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Ethnobotany of Yucatan
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Diss. ETH No. 13555
Yucatec Mayan Medicinal Plants:
Ethnobotany, Biological Evaluation
and
Phytochemical Study of Crossopetalum gaumeri
A dissertation submitted to the
SWISS FEDERAL INSTITUTE OF TECHNOLOGY ZURICH
For the degree of
Doctor of Natural Sciences
Presented by
ANITA SABINE ANKLI
Eidg. dipl. Apothekerin
born June 25, 1967
Zullwil/Meltingen (SO)
Accepted on the recommendation of
Prof. Dr. 0. Sticher, examiner
Prof. Dr. M. Heinrich, co-examiner
Dr. J. Heilmann, co-examiner
Zürich 2000
Acknowledgement
Acknowledgements
I wish to express my sincerest thanks to the following people:
- Prof. Dr. Otto Sticher, my supervisor, for giving me the chance to carry out a
ethnobotanical-phytochemical project in his research group, for providing
extraordinary working facilities as well as for the open door to discuss problems. I
am most grateful to Prof. Dr. Michael Heinrich, my co-supervisor, for introducing
me into the fascinating field of ethnobotany, for the encouraging discussions and
for the valuable support during the field study in Mexico. Special thanks go to Dr.
Jörg Heilmann for helping me with the structure elucidation of the isolated
compounds and accepting to support this thesis as a referee. -
- the healers and midwives of Chikindzonot, Ekpedz and Xcocmil for their
openness and patience in teaching me the usage of the medicinal plants and for
the opportunity to participate in their healing sessions and ceremonies. Many
thanks for answering my repeated questions. Only the knowledge of the healers
and midwives made it possible to carry out this thesis.
- the family of Don Abundio Chan Kauil and Dona Claudia Uc Cahun for their
hospitality, allowing me to stay in their home, for the delicious food and for the
introduction in the Mayan cosmovision. I would like to express my warmest thank
to Miriam and Gregoria Chan Uc for their courage to sleep in my house, thus
protecting me and for passing a good time together. I also like to thank Cresencio
Chan Uc for the uncountable discussions on any topic. (I like to apologize for the
mistakes I made due to cultural differences or lack of understanding).
- Marciana Poot Kauil for helping me as a Maya-Spanish translator and friend and
for making it easier to come in contact with the healers and midwives of Ekpedz
and for her help to collect plants in the forest and in hardly passable regions. I am
Acknowledgement
grateful to her father Don Silvestre Poot Poot and her mother for their hospitality
and the introduction in the actual Mayan beliefs and mystic tales.
- children of Chikindzonot who filled my house with great happiness and who
followed, not yet, the social roles and taboos.
- Dr. Ingrid Olmsted for the botanical support in the CICY (Centro de
Investigaciön Cientifica de Yucatan) and for the invitations to several botanical
excursions on the Peninsula of Yucatan. For the help of the botanical identification
I am very much indebted to the biologists and co-workers of the CICY, especially to
Jorge Carlos Trejo, Paulino Sima and Dr. Rafael Duran. I also like to thank the
botanists and specialists of the MEXU (Herbario Nacional de Mexico, Mexico D.F.):
Dr. Mario Sousa, Dr. Oswaldo Tellez, Dr. Rafael Lira, Dr. Jose-Luis Villasenor,
Dr. Fernando Chan and Dr. M. Martinez Gordilla.
- Dr. Ignacio Tuz and Dona Aurora (INI, Instituto Nacional Indigenista, Valladolid)
for supporting the project and inviting me to the meetings of the healers.
- Gisel Vargas, Roxana Chavarrfa and Don Julio Chavarrfa for their friendship
and hospitality in Mérida and Valladolid and for telling me stories about the
Mexican way of life.
- Dr. Barbara Pfeiler (Universidad Autönoma de Yucatan, Mérida) and Dr. Carlos
Viesca (UNAM, Universidad Nacional Autönoma de Mexico, Mexico D. F.) for the
linguistical and anthropological discussions and the invitations in their wonderful
homes. I am grateful to Dr. Ramon Arzâpalo (UNAM) and Jolanda Arzapâlo for
their fruitful discussions on Maya culture and for their hospitality in the biggest city
of the world.
Acknowledgement
- Dr. Carlos Zolla (INI, Mexico D.F.), Dr. Arturo Argueta and Dr. Gonzalo Solis
(INI, Mérida) for their advice to find a good place of study and for their friendliness
to accept and respect me as a broken-Spanish speaking person (at that time).
- Dr. Matthias Baltisberger (ETH) for sending the valuable equipment to dry
plants, Prof. Dr. Daniel Moerman (University of Michigan-Dearborn) for the
productive discussions about medicinal and non-medicinal plants.
- Dr. Oliver Zerbe for teaching me in the interpretation of NMR spectra and for the
untiring and fruitful discussions about NMR problems. I also wish to thank Dr.
Engelberg Zass for performing search on chemical compounds and Dr. Walter
Amrein, Oswaldo Greter and Rolf Häflinger for recording mass spectra.
- Dr. Hongmei Liu and Dr. Jimmy Orjala for their support and stimulating
discussions about chromatography and Micheal Wasescha for the determination
of KB cell cytotoxicity of plant extracts and pure compounds. My warm thanks go to
Dr. Barbara Frei Haller for the encouraging ethnobotanical discussions, the
literature search on plants and for her excellent pioneer work in ethnobotany in our
phytochemistry group.
- Prof. Dr. Horst Rimpler for the good co-operation of the Freiburg-Zürich group
(Albert-Ludwigs-University, Freiburg). I especially would like to thank Dr. Peter
Bork, Dr. Bilkis Heneka and Dr. Elke Beha.
- Dr. Lutz Wolfram, Dr. Peter Bauerfeind (University Hospital, Zurich), Dr.
Claudia Weiss, PD. Dr. Reto Brun, Cécile Schmid (Swiss Tropical Institute,
Basel), Regina Bruggisser (University, Basel), Jürg Gertsch (ETH) and Dr.
Helmut Wiedenfeld (University, Bonn) for testing extracts of different plants. I
would like to thank Dr. Christoph Schachtele and Dr. Frank Totzke for
determining protein kinase activity (Klinik für Tumorbiologie, Freiburg). I am
Acknowledgement
grateful to Prof. Dr. Marcus Schaub (University, Zurich) for proposing the
hypothesis about the effect of cardenolides in this treatment for snake bites.
- David McLaughlin and Anna Jen for the Engl.sh correction of this thesis.
- all my colleagues and staff at the Institute of Pharmaceutical Sciences, ETH
Zurich for the great time we had together, especially I like to thank my laboratory
colleagues (17L80).
- all my friends who visited me in my home in Yucatan and those who critically
supported my ideas and dreams.
Last but not least I express my deepest thank to my parents and Julian
Granados for their private support and patienc3, which gave me great motivation
to overcome various problems during this PhD-thesis.
Financial support during the work of this thesis was obtained from:
- Swiss Agency of Development Cooperation (SDC), Berne
- Swiss Academy of Natural Sciences (SANW), Zurich
- Barth Fonds of ETH, Zurich
Contents
Abbreviations
Summary
Resumen
Zusammenfassung
1
4
6
8
Part I Medicinal Ethnobotany
1 Introduction 12
1.1 Goal and objectives 16
2 Yucatan and the Mayas 17
2.1 Background 17
2.1.1 Geology and fauna 17
2.1.2 Ancient Maya history 18
2.1.3 From the Spanish conquest to the Caste War 21
2.1.4 The Yucatec Maya today 23
2.1.5 Manuscripts on Maya medicinal plants 28
2.1.6 Medicinal ethnobotany of Yucatan 30
2.2 Methods in the field 33
2.3 Abbreviations mentioned in the plant list 37
2.4 Plant list 41
2.5 Informants 60
2.6 Gardens of medicinal plants 61
2.7 Selection of plant species for their biological evaluations 62
3 Publication I: Medical Ethnobotany of the Yucatec Maya:
Healer's consensus as a quantitative criterion 63
4 Publication II: Yucatec Maya medicinal plants versus non-
medicinal plants: indigenous characterization
and selection 97
Part II Plant Evaluation
5 Publication III: Yucatec Mayan medicinal plants:
Evaluation based on indigenous uses 134
6 Additional results 160
6.1 Antimicrobial activity 160
6.2 Comparison of disk method and TLC method 161
6.3 Protein kinase activity; Method and results 162
6.4 Other activities 167
6.5 Crossopetalum gaumeri- the plant species for the
phytochemical study 168
Part III Phytochemistry of Crossopetalum gaumeri
7 Celastraceae family and the genus Crossopetalum 170
7.1 Botanical taxonomy 170
7.1.1 Crossopetalum gaumeri 171
7.2 Phytochemistry of the Celastraceae 172
7.2.1 Terpenoids (terpenes, isoprenoids) 172
7.2.2 Alkaloids 176
7.2.3 Flavonoids and other phenolic compounds 176
7.3 Phytochemistry of Crossopetalum species 176
7.4 Biosynthesis of terpenoids (terpenes, isoprenoids) 177
7.5 Chemosystematic and phylogenetic relationships 180
7.6 Biological activities among the Celastraceae 182
7.7 Popular medicinal use 185
7.7.1 Yucatec Maya medicinal use of C gaumeri 185
7.7.2 Medicinal application of other Crossopetalum species 186
7.7.3 Global medicinal use of Celastraceae species 186
8 Methods (isolation procedure) 188
8.1 Thin layer chromatography (TLC) 188
8.2 Vacuum liquid chromatography (VLC) 188
8.3 Middle pressure liquid chromatography (MPLC) 188
8.4 High performance liquid chromatography (HPLC) 189
8.5 Open column chromatography 189
8.6 Liquid-liquid partition (LLP) 189
9 Methods of structure elucidation 190
9.1 Nuclear magnetic resonance spectroscopy (NMR) 191
9.2 Mass spectrometry (MS) 193
9.3 UV spectroscopy (UV) 194
9.4 Optical rotation 194
9.5 Acidic hydrolysis 194
10 Plant extraction 195
10.1 Small scale plant extraction 195
10.2 Large scale plant extraction 195
10.3 Fractionation of the methanol extract 196
10.4 Fractionation of the dichloromethane extract 197
11 Structure elucidation of the isolated compounds 200
11.1 Cardenolides 200
11.2 Ourateacatechin 213
11.3 Triterpenes 219
11.3.1 Pristimerin 219
11.3.2 Friedelane-3-on-29-ol 221
11.3.3 2,3,7-Trihydroxy-6-oxo-1,3,5(10),7-tetraene-24-nor-
friedelane-29-oic acid methylester 224
11.3.4 Celastrol 228
11.4 3,15-Dihydroxy-18-norabieta-3,8,11,13-tetraene 231
12 Biological activities of isolated compounds 233
12.1 Cytotoxicity 233
12.2 Other activities 235
13 Biomedicine, a way to explain the medicinal use
of C. gaumeri ? 236
13.1 Gastrointestinal problems 236
13.2 Snakebites 237
13.2.1 C. gaumeri used in the treatment for snake bites 238
13.2.2 What have the cardenolides to do with snake bites ? 240
14 Publication IV: Cytotoxic cardenolides and antibacterial
terpenoids from Crossopetalum gaumeri 243
15 Conclusion 261
References 264
List of publications 278
List of poster presentations 278
Oral presentations 279
Curriculum Vitae 280
Abbreviations
Abbreviations
A
AT
[a]D
B
C
ATCC
CDCI3
CHCI3
CH2CI2
Ô
d
dd
2D
DEPT
DER
DQF-COSY
EI-MS
ESI
eV
EYE
FAB-MS
FEM
Gl
H20
HMBC
HSQC
HPLC
non-polar extract (dichloromethane-methanol 2:1)
bites and stings of venomous animals
specific optical rotation
polar extract (1-butanol)
ethanol extract
American type cultures collection
deuterated chloroform
chloroform
dichloromethane
chemical shift
doublet
double doublet
two-dimensional
distortionless enhancement by polarization transfer
dermatological conditions
double quantum filter correlation spectroscopy
electron impact-mass spectrometry
electronspray ionization
electron Volt
illnesses of the eyes
fast atom bombardment-mass spectrometry
women's medicine
gastrointestinal disorders
water
heteronuclear multiple bond correlation
heteronuclear single quantum correlation
high performance liquid chromatography
1
Abbreviations
Hz
'Cgo
INADEQUATE
J
KB cell line
m
MeOD
MeOH
MIC
MHz
MS
MPLC
m/z
Mr
n-BuOH
NCI
NF-kB
NMR
3-NOBA
NOE
OTH
PK
PFE
ppm
q
RES
ROESY
Rf
Hertz
50 % inhibition concentration
incredible natural abundance double quantum transfer
experiment
coupling constant
human nasopharyngeal carcinoma cell line
multiplet
deuterated methanol
methanol
minimum inhibition concentration
Megahertz
mass spectrometry
medium pressure liquid chromatography
mass to charge ratio (MS)
relative mass
1-butanol
National Cancer Institute
nuclear factor kB
nuclear magnetic resonance
3-nitrobenzyl alcohol
nuclear Overhauser effect
other uses
protein kinase
illnesses associated with pain and fever
parts per million
quartet
respiratory illnesses
rotating frame Overhauser enhancement spectroscopy
retention factor (TLC analysis)
RP reversed phase
RT room temperature
s singulet
Si gel silica gel
sp. species
spp. species (plural)
ssp. subspecies
t triplet
TLC thin layer chromatography
TOCSY total correlation spectroscopy
UR urological problems
UV ultraviolet spectroscopy
VLC vacuum liquid chromatography
Summary
Summary
The use of medicinal plants played an important role in the lives of the Ancient
Maya. Also today, more than 450 years after the conquest of the New World,
medicinal plants are an essential part of the medical system of the lowland Maya of
Yucatan.
During 18 months of field work in three Yucatec Mayan communities (Mexico)
information about medicinal plants, the concepts of disease and methods of
treatment were collected. Based on the knowledge of 40 healers and midwives,
360 medicinal plants and 1828 single- use reports could be documented. In a
quantitative approach, the most frequent illnesses of this region were evaluated.
Gastrointestinal problems (32 %) and dermatological conditions (19 %) were the
most important medical problems, followed by illnesses associated with pain and/or
fever (13 %), respiratory illnesses (11 %), '"women's medicine" (8 %), other uses (5
%), bites and stings of venomous animals (5 %), urological problems (4 %) and
eye disorders (3 %). To better understand the selection criteria for medicinal
plants, 12 healers and midwives were interviewed about ten plants that in their
opinion have no medicinal value. The characteristics of these non-medicinal plants
were compared with those of the medicinal ones. The results showed that odor and
taste are essential criteria for plant characterization. Also humoral classification
plays an important role. In general, illnesses are classified as hot or cold and the
medicinal plants ought to have the opposite humoral classification. Color, form and
texture are also important criteria in the selection of medicinal plants.
In the second part of the study, 48 medicinal plants were evaluated in several
bioassays. All plant extracts were tested for their antibacterial (gram-negative and
gram-positive bacteria), cytotoxic (KB cells) and anti-inflammatory (NF-kB) activity.
In addition, they were tested in further bioassays based on their indigenous uses.
Plant species used against gastrointestinal problems were evaluated for
antiparasitic (Giardia duodenalis) and additional antibacterial (Helicobacter pylori
4
Summary
and Campylobacter jejuni) activity. The plants of the group used for skin conditions
were also tested for their anti-fungal effects (Candida albicans). For the plants
traditionally used against pain and fever the antimalarial activity (Plasmodium
falciparum) was examined. Plants used in the treatment of type II diabetes were
tested for a-amylase inhibitory effect and the dopamine D2 receptor test was
applied for the taxa used in the group "women's medicine". Different activities were
evaluated that substantiate the traditional use of the herbal remedies.
In a third step, one plant species -Crossopetalum gaumeri (Celastraceae)- was
investigated phytochemically. The roots of this plant were chosen due to their oral
and local use against diarrhea and snake bites, and on the basis of the positive
results obtained in the above mentioned bioassays. From the methanol extract one
known and four new highly cytotoxic cardenolides and the known ourateacatechin
were isolated. The dichloromethane extract afforded a new diterpene of the
abietane type and a new pentacyclic triterpene. Three known triterpenes
(pristimerin, celastrol and friedelane-3-on-29-ol) were also isolated and examined
in different bioassays. Pristimerin and celastrol showed high antibacterial activity
and remarkable cytotoxicity against KB cells. In some respects, the activities of the
isolated compounds substantiate the indigenous uses of C. gaumeri. However, the
plant should be used with caution due to its high cytotoxicity.
5
Resumen
Resumen
El uso de las plantas médicinales por las sociedades mayas prehispanicas
representaba parte importante del sistema medicinal. Aün hoy en dia, después de
450 ahos de la conquista, las plantas médicinales son una parte esencial del
conocimiento médico-farmacolôgico de las sociedades mayas en la Peninsula de
Yucatan.
Se trabajö durante 18 meses en très comunidades mayas, colectando informacion
sobre el uso de las plantas médicinales, tratamiento de las enfermedades y la
conceptualizacion de lo que se denomina enfermedad. Basado en el conocimiento
de 40 curanderos y comadronas se documentaron 360 especies de plantas
médicinales y 1828 usos médicinales. Usando un criterio cuantitativo se
clasificaron las enfermedades mâs frecuentes de la region en que se trabajö. Los
problemas gastrointestinales (32 %) y las enfermedades de la piel (19 %) son los
principales problemas de salud. Seguidamente se encuentran las enfermedades
relacionadas con problemas de dolor y/o fiebre (13 %), problemas respiratorios (11
%), medicina de las mujeres (8 %), otras indicationes (5 %), mordedura y picadura
de animales venenoso (5 %), problemas urologicos (4 %) y enfermedades de la
vîsta (3 %). Para entender cuâl es el criterio que se usa para decidir si una planta
es medicinal o no, se encuestö a 12 curanderos y comadronas, a los cuales se les
pidio que seleccionasen 10 especies de plantas no médicinales. Basandose en
dichas especies se les preguntö sobre el criterio de selecciön para dichas plantas.
Las respuestas acerca de las plantas no médicinales se compararon con las
obtenidas para las plantas médicinales. Los resultados muestran que el sabor y el
olor son caracteristicas para las selecciön de una planta medicinal. Las
caracteristicas humorales (frîo o caliente) son también otro criterio de importancia
para la selecciön de las plantas médicinales. En general las enfermedades se
clasifican en frias y calientes y este criterio se considéra en la clasificacion
humoral de las plantas. Color, forma y textura son también criteriorios de
selecciön de gran significado para la selecciön de una planta medicinal.
6
Resumen
La segunda parte del trabajö consistiö en la evaluaciön de 48 especies de plantas,
usando diferentes bioensayos. Todas los extractos de plantas fuero evaluados con
bioensayos antibacteriales (Bacterias gram-positivas/negativas), citotixicidad
(Celulas KB) y efecto antiinflamatorio (NF-kB). Ademâs, se testaron los usos
indigenas, usando otros bioensayos. Plantas que se usan contra enfermedades
gastrointestinales, se probaron contra usando bioensayos antiparasitales {Giardia
duodenalis) y antibacteriales {Helicobacter pylori, Campylobacter jejuni ). Las
plantas clasificadas en el grupo de enfermedades de la piel, fueron testadas contra
actividad antifungicida {Candida albicans). Las plantas clasificadas como
antifebriles, fueron testadas contra malaria, usando Plasmodium falciparum .Las
plantas clasificadas como Diabetes II se testaron usando el test a-Amylase contra
hiperglucemia. Se encontraron respuestas activas en las plantas, que
corresponden a los usos indigenas en que se clasificaron las plantas.
En la tercera parte del estudio se analisö fitoquimicamente la especie
Crossopetalum gaumeri. Las rai'ces de esta especie son de uso antidiarréico y
antiviperino, tal y como lo confirman los resultados del bioensayo en que se testé
dicha actividad. Del extracto obtenido de la planta con metanol, se encontraron
cuatro nuevos glucösidos cardîacos, altamente citotöxicos; asi como también el
conocido compuesto Ourateacatequina. A partir del extracto con diclorometanol,
se encontaron nuevos abitandipertenos y nuevos pentacicloterpenos. Asi mismo,
se aislaron très terpenos ya conocidos (Pristimerin, Celastrol und Friedelane-3-on-
29-oI) y fueron testados usando los bioensayos anteriormente mencionados. La
Pristimerina y el Celastrol muestran una gran capacidad antibacterial y citotöxica.
Desde una amplia perspectiva se puede decir, que el uso de las plantas
médicinales aqui estudiadas, dada su alta citotoxicidad, debe recomendarse de
una forma cautelosa.
7
Zusammenfassung
Zusammenfassung
Der Gebrauch von Medizinalpflanzen spielte im Leben der alten Maya eine
wichtige Rolle. Auch heute, nach über 450 Jahren der Eroberung der Neuen Welt,
sind Medizinalpflanzen ein wesentlicher Bestandteil des Gesundheitswesens der
Tiefland-Maya von Yukatan.
Während eines 18-monatigen ethnobotanischen Feldaufenthaltes in drei
yukatekischen Maya-Dörfern (Mexiko) wurden Informationen über Medizinal¬
pflanzen, Krankheitskonzepte und Behandlungsmethoden gesammelt. Auf das
Wissen von 40 Heilem und Hebammen basierend konnten 360 Medizinalpflanzen
und 1828 einzelne Anwendungen dokumentiert werden. In einer quantitativen
Auswertung wurden die häufigsten Erkrankungen dieser Region eruiert.
Gastrointestinale Beschwerden (32 %) und Hauterkrankungen (19 %) waren die
wichtigsten medizinischen Probleme, gefolgt von Krankheiten verbunden mit
Schmerz und/oder Fieber (13 %), respiratorischen Beschwerden (11 %),
„Frauenmedizin" (8 %), andere Indikationen (5 %), Bisse und Stiche von Gifttieren
(5 %), urologische Probleme (4 %) und Augenkrankheiten (3 %). Zum besseren
Verständnis der Auswahlkriterien von Medizinalpflanzen wurden 12 Heiler und
Hebammen über zehn Pflanzen, welche ihrer Meinung nach keinen medizinischen
Wert haben, befragt. Die Eigenschaften dieser Nicht-medizinalpflanzen wurden mit
denjenigen der Medizinalpflanzen verglichen. Die Resultate zeigten, dass Geruch
und Geschmack wichtige Parameter zur Charakterisierung der Pflanzen sind. Auch
die humorale Einteilung spielt eine wichtige Rolle. Im allgemeinen werden die
Krankheiten in heiss und kalt eingeteilt und die verwendeten Medizinalpflanzen
erhalten die entgegengesetzte humorale Bezeichnung. Farbe, Form und Textur
sind bei der Auswahl von Medizinalpflanzen ebenfalls bedeutende Kriterien.
Im zweiten Teil der Arbeit, wurden 48 Pflanzenarten in verschiedenen biologischen
Testsystemen evaluiert. Alle Pflanzenextrakte wurden auf antibakterielle (gram¬
negative und gram-positive Bakterien), zytotoxische (KB Zellen) und entzündungs¬
hemmende (NF-kB) Wirkung untersucht. Zusätzlich wurden sie bezüglich der
8
Zusammenfassung
indigenen Anwendung in weiteren Testsystemen geprüft. Pflanzen, die gegen
gastrointestinale Beschwerden eingesetzt werden, wurden auf antiparasitäre
(Giardia duodenalis) und zusätzliche antibakterielle (Helicobacter pylori,
Campylobacterjejuni) Aktivität untersucht. Pflanzen der Gruppe Hauterkrankungen
wurden ferner auf antifungale Wirkung (Candida albicans) getestet. Die
Antimalaria-Aktivität (Plasmodium falciparum) wurde für die Pflanzen, die gegen
Fieber angewendet werden, ermittelt. Pflanzen zur Behandlung von Diabetes II
wurden in einem a-Amylase Test auf antihyperglykämische Wirkung geprüft und
der D2 Dopamin-Rezeptor Test wurde für die Spezies, die im Gebiet der
Frauenmedizin eingesetzt werden, verwendet. Es resultierten verschiedene
Wirkungen, welche die traditionelle Verwendung der pflanzlichen Heilmittel
begründen können.
Im dritten Teil der Studie, wurde eine Pflanzenart -Crossopetalum gaumeri
(Celastraceae)- phytochemisch untersucht. Die Wurzeln dieser Pflanze wurden
auf Grund der oralen und lokalen Anwendung gegen Diarrhöe und Schlangenbisse
sowie der positiven Resultate in den oben genannten Testsystemen ausgewählt.
Aus dem Methanolextrakt konnten ein bekanntes und vier neue, sehr zytotoxische
Cardenolide sowie das bekannte Ourateacatechin isoliert werden. Die
Untersuchung des Dichlormethanextraktes führte zu einem neuen Abietanditerpen
und zu einem neuen pentazyklischen Triterpen. Zusätzlich wurden drei bekannte
Terpene (Pristimerin, Celastrol und Friedelan-3-on-29-ol) isoliert, und in den
verschiedenen biologischen Testsystemen geprüft. Pristimerin und Celastrol
zeigten starke antibakterielle Wirkung sowie beachtliche Zytotoxizität in KB Zellen.
In mancher Hinsicht belegen die Aktivitäten der isolierten Substanzen die
traditionelle Verwendung, doch sollte die Pflanze auf Grund der starken
Zytotoxizität mit Vorsicht eingesetzt werden.
9
Introduction
1 Introduction
The main goal of this interdisciplinary study is the detailed documentation of the
medicinal plants of the Yucatec Maya of three communities and their botanical
identification. The understanding of the healing methods and the concepts of
diseases were further purposes of the field study of 18 months. In the second
stage, the most important plant species were evaluated in various bioassays based
on indigenous uses. Additionally, one of the species was investigated
phytochemically and the isolated compounds were examined in different
bioassays.
Medicinal plants are an important element of the medical system of the Yucatec
Maya (Mexico). An impressive number of healers and midwives apply the empirical
medicine, which has been developed over hundreds of years. Some plant uses are
supported by documentary evidence in old manuscripts written by the Maya or
Spaniards in the 16th century (Arzapälo Marin, 1995; Diego de Landa, 1992). Also
the performance of ceremonies addressing the rain-god to ask for rain and the
ceremonies for protecting milpas are examples of cultural heritages which go back
to the time of the Ancient Maya (Diego de Landa, 1992). The Yucatec Maya are
one group of the direct descendents of the Ancient Maya, which were a highly
advanced civilization. Herbal remedies must have been very important to the them.
Since no historic records survived from this it is of particular interest to study the
use of medicinal plants by the modern Maya. Another important point for studying
the medical system of the Yucatec Maya is the lack of modern studies of medicinal
plants and their scientific identification. There are several interesting books and
studies about medicinal plants of Yucatan. However, their information is often
based on secondary sources with data documented at earlier times or with plants
not scientifically identified.
12
Introduction
Ethnobotany is an interdisciplinary specialty, which studies plant-human
interrelationships, which occurs in different aspects of the lives of human beings,
such as medicine, nutrition, and ecology. It also includes various fields such as
ethnology, botany, medicine, linguistics and pharmacy. Knowledge about useful
plants must go back to the beginning of human existence. Humans certainly had to
differentiate between plants without any use, plants from which they could obtain
nourishment or stimulation, plants which could alleviate ailments or even cure
sicknesses, plants with psychoactive properties, and plants which could be used to
kill (Schultes and Siri von Reis, 1995).
Medical ethnobotany is an interdisciplinary science, which studies the use of
medicinal plants among cultures. There are several ways to examine this topic.
One is the anthropological approach to the way indigenous people interpret and
treat their useful plants. Another goal concerns the documentation of the medicinal
plants and their evaluation in subsequent bioassays. The discovery of natural
products for the development of new drugs is a further aim in this field of study.
The term "medical ethnobotany" is closely allied with ethnopharmacology, however
it does not necessarily include the examination of the physiological and clinical
impact of plant use on human health.
One of the objectives of medical ethnobotany is to document the uses of medicinal
plants and thus to rescue an important cultural heritage. The quantification of
documented medicinal plant usages determines which plants are most frequently
used in a culture. Their evaluation in biological test systems and their detailed
study are of high importance for the safety and efficacy of plants used in primary
health care. The WHO (World Health Organization) has estimated that about 80 %
of the people living in less developed countries rely almost exclusively on
traditional medicine for their primary health care needs. Thus, there is a need for
the study of these plants concerning safety and efficacy and to develop galenical
preparations that are standardized and stable (Farnsworth, 1980). The goals of
WHO are: (1) To strengthen research on the safety and efficacy of herbal
13
Introduction
medicines. (2) To strengthen and promote the rational use of herbal medicines
(WHO, 1993).
The phytochemical study of medicinal plants based on an ethnobotanical
approach led to the development of several drugs used in modem medicine. One
of the best examples is curare (from Chondrodendron spp., Menispermaceae). The
paralyzing effect of the plant extract is used as an arrow-poison among some tribes
in South America. One component of this plant extract, tubocurarine, is used as a
muscle-relaxant in modern medicine. Another example is foxglove {Digitalis
purpurea, Scrophulariaceae) which was used in England to treat dropsy and
epilepsy. Today the cardiac glycosides, digitoxin and digoxin, as the most
important compounds of the species are used in the treatment of chronic heart
insufficiency. One of the most recent and promising medicines is artemisinin,
isolated from Artemisia annua (Asteraceae), a species which has been used in
traditional Chinese medicine against fever and malaria for two thousand years.
This antimalarial drug is of great interest due to the development of resistance
against the known remedies and the great health risk in tropical areas. Farnsworth
et al. (1985) mentioned a total of 119 pure compounds, isolated from plants which
are currently used in medicine.
The phytochemical investigation of plants, the isolation of compounds and their
examination in biological test systems not only is essential for the study of safety
and efficacy of plants but can also help to find potent and selective drugs. There is
considerable need to obtain new, active compounds particularly in the field of
cancer, infections and tropical diseases. The plant world with about 250,000
species of flowering plants is an immense source of chemical compounds with a
vast array of unusual chemical structures that display a variety of biological
activities. Until now only a small part of this resource has been explored.
The anthropological approach of medicinal ethnobotany investigates, among
other things, the selection criteria for medicinal plants and the classification system
14
Introduction
of plants used among indigenous people. Some aspects of this key field of study
are reflected in the following questions. Why is a plant medicinal? What are the
criteria for medicinal plant selection? How do indigenous people classify plants?
Are non-medicinal plants also classified? What are the differences between
medicinal and non-medicinal plants?
Several studies have systematically investigated the hot-cold concept and its role
in indigenous medical systems especially for classifying plants and illnesses
(Foster, 1988; Ingham, 1970). Some authors mentioned the hot-cold categorization
of humoral medicine as the "basic cognitive principle" of traditional medicine in
Latin America (Tedlock, 1987). Others criticized the hot-cold system as too narrow
to explain plant choices and showed that taste and odor are important parameters
for the characterization of medicinal plants (Brett, 1992; Heinrich, 1989). To the
author's knowledge, there has been no study that focused on the comparison of
non-medicinal plants with medicinal ones and on the ways people perceive such
plants. Thus, the study of the non-medicinal plants could shed new light on
indigenous selection criteria. Furthermore it helps us to better understand the
classification system of a culture. Hence, one of our studies focused on the
Yucatec Maya medicinal plants in comparison with the non-medicinal ones
concerning taste and smell perception as well as the hot-cold concept.
15
Introduction
1.1 Goal and objectives
The goal of the studies presented in this thesis was to document the medicinal
plants of the Yucatec Maya; to study their perception and classification of the
medicinal plants; to evaluate the most important species in different bioassays; to
phytochemically study one species and to examine the isolated compounds for
biological activity.
Specific Objectives
To understand the Mayan healing practice, the illnesses, their cause, symptoms
and prevention; the medicinal plants and the preparation of the remedies, the
combination with other plants, the popular plant name, the plant part used, the
dosage of the remedies and side effects as well as their classification.
To obtain knowledge about the history of becoming a healer, the way they select
medicinal plants, and how they diagnose illnesses and perform healing
ceremonies.
To evaluate important medicinal plants in different bioassays based on the
indigenous use.
To select a medicinal plant which is used against the most important health
problem for phytochemical investigation.
To study a plant species phytochemically using bioactivity-guided isolation and
isolate pure compounds, to identify the chemical nature of the compounds, to
examine their activity and cytotoxicity as well as to correlate them with the
indigenous use of this plant.
To return the documented data and the results of this thesis to the informants of
the study region and the people and organizations which supported this project
as well as to the libraries of Mérida so that the thesis is accessible to people
who have an interest in this subject.
16
Medicinal Ethnobotany
2 Yucatan and the Mayas
2.1 Background
2.1.1 Geology and fauna
The Maya territory occupies the northwestern half of the Central American Isthmus
and is divided into highlands and lowlands. The proximity of two coasts, the
contrast of rainfall due to the tropical climate, and the varied relief result in
considerable differences in the two environments.
The Yucatan Peninsula consists of the Mexican states of Yucatan, Campeche,
Quintana Roo, as well as small portions of the states of Chiapas (the Lacandon
Jungle and Marquez de Comillas) and Tabasco (Balancan region), the north of
Belize and the Peten region of Guatemala. The Maya Mountains constitute the
southeastern limit of the Peninsula, and the mountains to the north of Chiapas form
the southwestern limits. The Mexican part of the Peninsula is formed by a porous
limestone platform with altitudes of less than 350 m above sea level. Due to the
porous limestone the rainwater sinks immediately below the surface, where it forms
underground reservoirs in great caves. Due to the absence of surface streams the
cenotes (open sinkholes, which are connected to the water-bearing bed) have
always been the main sources of drinking water for the inhabitants. Rivers only
exist in the extreme southwest and southeast.
The Yucatan Peninsula maintains a very characteristic flora. Of the 2,300 species
of vascular plants, which build the flora of the Mexican part of the Peninsula, 168
are endemic taxa (7.3 %) (Düren et al., 1998). The vegetation shows floristic
elements of the neighboring areas such as the Antillean region, Central America,
and the southeast of Mexico (Standley, 1930; Rzedowski, 1988). According to
Estrada-Loera (1991) the most important floristic elements are those of Central
America, however the endemic species and the floristic similarities with the Antilles
are of special interest (Standley, 1930).
17
Medicinal Ethnobotany
Not only the geological and floristic aspect, but also by its physiographic feature
and human inhabitants, the Yucatan Peninsula is sharply differentiated from the
rest of Mexico.
Most of the Peninsula shows an Aw climate (hot with long dry period, rainy season
in summer), however a narrow northern coastal strip is of BS type (dry and hot).
More information of the geographical, floristic and climatic conditions is provided, in
publication I.
2.1.2 Ancient Maya history
Origins of agriculture
12,000 years ago human population subsisted by various forms of hunting and
gathering. In Mesoamerica, a shift toward plant and animal domestication occurred
9,000 years ago. In the Fertile Crescent of the Near East it happened 10,000 years
ago and in Southeast Asia 7,000 years ago (Lewin, 1999). Social and political
complexity was a consequence of the adoption of agriculture.
The Preclassic Period (2,000 B.C.-A.D. 250)
By about 1,500 B.C. settled farming life was established in most parts of Meso¬
america, including the Maya world. Before 1,000 B.C., a new kind of society was
emerging along the Gulf Coast: the Olmec centers. The society stood out for the
sharp contrast and status, marked with centralized political power reflected in
monumental architecture and sculpture. Although some parts of the Maya world,
mostly in the highlands, were tied into the economic networks of the Olmec world.
The early Maya communities in the lowlands were small and simple villages. After
500 B.C. some communities were beginning to reflect a new development. Jewelry
and other goods made from exotic raw material indicated increasing prosperity,
and sharper differences in wealth and social status. Decorated public buildings
reflected the emergence of powerful permanent leader, chiefs or kings (Henderson,
1997).
18
xvï r???
Figure 2.1. Landscape of the Yucatan Peninsula with Uxmal
Figure 2.2. Cenote (natural sinkhole)
19
Medicinal Ethnobotany
The Classic Maya Civilization (A.D. 250-1000)
Especially during the Early Classic period, the concentration of the political and
economic power in the hands of elite grew. Several regions experienced intensified
population growth with well-developed hierarchies of communities. Many cities
enjoyed a boom in building. Relationships with distant societies intensified. Nobles
acquired greater political and religious authority. Specialist multiplied in every field:
architecture, arts, crafts, writing and in the intellectual sphere generally.
Interchanges took place in the aristocratic as well as in the intellectual sphere
among priests devoted to astronomical and astrological investigations. The Classic
period was a time of cultural florescence throughout the Maya world (Henderson,
1997).
Transformation. In the ninth century, new processes, involving internal strains as
well as external pressures disrupted long-standing patterns of growth and
expansion. This episode, which was once conceptualized as a sudden and
universal collapse of all facets of the Maya civilization, now appears to have been a
series of processes that operated over several centuries. An extraordinary and
unusual aspect of the transformation was that most regions went through a stage
of deep decline in the cultural development at about the same time (Henderson,
1997). By the middle of the tenth century quite every southern city was an
abandoned ruin. The state institutions declined with consequent transformations of
aristocratic economies. In most regions, village and household life went on, but in
much-simplified political and economic systems. The cities in northern Yucatan
generally continued to flourish during the tenth century (Chichén Itzâ, Uxmal),
although a process of decline may have begun in a few places as external
pressures intensified (Henderson, 1997).
The Postclassic Maya (A.D. 1000-1525)
During the Postclassic period long-distance exchange increased and agriculture
was less central to economic systems. Political systems shifted away from a near-
exclusive focus on one person as the ruler, to new, more flexible forms of
20
Medicinal Ethnobotany
organization that involved a much broader distribution of power. The public roles of
religion were reduced but ritual maintained a central place in domestic life
(Henderson, 1997).
The Lowlands. When Chichén Itzâ fell into decline as a political and economic
center, Mayapän, an important political center, replaced it. The Mayapân's sphere
disintegrated into different provinces at least a century before the arrival of the
Spaniards. At that time, several families controlled the provinces in Yucatan: for
example Canul (West of Yucatan), Cupul (eastcentral Yucatan), Tutul Xiu (Mam)
and Cocom (Sotuta) (Henderson, 1997).
2.1.3 From the Spanish conquest to the Caste War
The Spanish conquest (16th-17th centuries)
In 1517 and 1518 two Spanish expeditions in search of gold, new territory and
slaves were started from Cuba. In 1519 Hernân Cortés landed in Veracruz from
where the Spaniards conquered the Aztecs, situated in the center of Mexico, in a
year. It took another 20 years to conquer the numerous Maya provinces of
Yucatan. In Yucatan and Guatemala the colonial regime was firmly established
with Montejo (about 1546) and Alvarado (about 1525) , respectively. However, until
about 1697, vast areas of jungle between the mountains of Guatemala and
northern Yucatan remained unconquered.
The colonial regime
Under Spanish law the Indians were considered subjects of the Spanish crown and
slavery was outlawed. As well as being slaughtered during the Conquest itself, the
Indians also fell victim to viral and other infections transmitted by the Spaniards,
which decimated the population. The increase of the power of the Catholic Church
and the suppression of ancient beliefs provoked rebellions among the Mayas,
including those of Canek (Yucatan) in 1671, Chiapas (1692), and the Tzeltal
rebellion at Cancuc in 1713. Fray Diego de Landa ordered an auto-da-fé during
21
Medicinal Ethnobotany
which Indians were tortured and executed and hundreds of idols and more than
twenty Mayan codices (books) were burnt (Baudez and Picasso, 1990).
Only four codices of the pre-conquest Maya culture written in hieroglyphs could be
rescued and are known today. All of them are made of amate (bark of Ficus sp.)
and have 12-56 pages (9-25 x 12-20 cm). The Codex Dresden conserved in the
Sächsische Landesbibliothek of Dresden (Germany) mentions mythology,
astrology, ceremonies and Gods. The content of the Codex Madrid or Tro-
Cortesianus, in the Museo de Americas of Madrid, refers principally to the
prophecy with the following themes: hunting, agriculture, cloth and rituals asking for
rain. The Codex Paris can be found in the Bibliothèque Nacional de Paris contains
sequences of ceremonies and rites. Codex Grolier is conserved in the Instituto
Nacional de Antropologia e Historia (Franch, 1992).
After independence
The Mexican priest Miguel Hidalgo initiated the call for independence from Spain.
After a long struggle, Mexico declared independence in 1821, followed immediately
by Yucatan, Chiapas and Gutemala. In 1841 Yucatan declared independence from
Mexico.
Caste War (1847-1904)
During the second half of the 19th century the Yucatan Peninsula was shaken by a
violent conflict between the Maya and Europeans. The Indians, heavily taxed by
the government, saw their ancestral lands being taken from them. The Maya,
armed by English settlers in Belize, regained 90 % of their lands. At this time the
Mayas inexplicably withdrew to their villages. It is said that the Mayas went home,
because of the beginning of the rainy season, thus they had to plant their milpa.
The Europeans regrouped with the help of military force of Mexico and the United
States. Hence, the Maya were driven back to Chan Santa Cruz (now Felipe Carrillo
Puerto), where they founded the cult of the Talking Cross. Inspired by the oracle
that promised them victory, they resisted vigorously for several years. In 1904, the
22
Medicinal Ethnobotany
eastern part of the Peninsula (Quintana Roo) converted to Mexican national
territory. After 1920, the chicle production and chicle gum, as its main product
{Manilkara zapota L. Van Royen), as well as the export of mahagony {Swietenia
macrophylla King) in the forest of Quintana Roo attracted people from different
parts of the country. The rebelling Maya had their own territory and did not permit
the entry to strangers. In 1936, this Maya region was accepted by the Government
as a Maya zone.
2.1.4 The Yucatec Maya today
Today more than three million people speak one of the twenty-six Mayan
languages and dialects, which are divided into ten large groups. Most of these
different languages are spoken in Guatemala, just a few ones are spoken in
Mexico. The languages are classified into the following groups (Castaheda, 1988;
Coe, 1975): (1) Huastec, Chicomuceltec, (2) Choi, Chontal, Chorti, Mopan, (3)
Tzeltal, Tzozil, Tojolabal, (4) Chuj, (5) Kanjobal, Jacaltec, Solomec, (6)
Motozintlec, (7) Mam, Tec, Aguatecpec, Ixil, (8) Quiche, Rabinalachi, Uspantec,
Cakchiquel, Tzutuhil, (9) Kekchi, Pocomchi, Pocoman, (10) Yucatec (dialects:
Lacandon, Itza)
On the Peninsula one-third of the population (1.1 million) have Yucatec Maya as
their mother tongue (Wilhelmy, 1990). Even though the Maya population increased
since 1950 this group has suffered a decline in the relative number of the native
speakers (Olivera et al., 1982). Bilingual programs at schools have always had the
goal of assimilation and resulted in a move towards learning Spanish, but this goal
seems to have come under attack because of ideological changes in education in
Mexico and autonomous movements of indigenous people throughout the
Americas (Burns, 1998). Land problems in the Maya area are still not resolved.
Two insurrections, one in Guatemala (1963-1993) and one in Chiapas (1994-...)
are evidence of the multitude of problems that still persist.
23
Medicinal Ethnobotany
Figure 2.3. Groups of Mayan languages (Coe, 1975)
Chikindzonot, Ekpedz, Xcocmil - the study area
According to oral tradition Chikindzonot (= west of the cenote; sinkhole) was found
in the 14th century. However, until the end of the 16th century a stable encomienda
(place of forced labor) was known. Chikindzonot and the neighboring villages
Ekpedz and Xcocmil were abandoned during the Caste War and repopulated at
around 1915. The inhabitants of the villages still tell stories about the outbreak of
the war in Thiosuco, a nearby village, and the recovery and rebuilding of the
villages afterwards. Oversized churches dominate Chikindzonot and Ekpedz, this is
a common picture in most villages in Yucatan. The walls of the catholic churches
and the baptismal font are ornamented with indigenous figures. This kind of fusion
of Maya culture with Christian and Spanish elements can be seen in practically
every part of daily life. For example, the inhabitants of Yucatan celebrate two
24
Medicinal Ethnobotany
baptisms, a Christian one and hekmetz, which is a heritage of the Ancient Maya
(Diego de Landa, 1990). Chocolate (formerly cacao) is an engagement gift, which
the fiancé gives to the family of his future wife. As opposed to this, the bride at the
official wedding celebration in the church wears a white wedding dress of
European style. The architecture of the houses made of planks, branches and
palm leaves looks the same as 500 years ago. However, the building of stone
houses is getting more popular. Food consists mainly of corn products, beans,
marrow and chili. To enrich the meal, animals are hunted in the forest or domestic
animals like chicken, pigs, and cows, introduced by the Spaniards, are kept. The
work of the women generally takes place in the house and the extensive home
garden. The most important site in the kitchen is the fireplace which is always built
using three stones. A small table and chairs are used as working place for the
women, to make tortillas, and as the dining-table for the whole family. The metate
(millstone), formerly used to turn the corn into flour, has been replaced by a hand
or motor-operated mill. Today the metate is used for grinding herbs to enrich meals
or for the preparation of medicine. Gourds and bowls made of clay or wood as well
as metal and plastic bowls are common utensils in the kitchen. Men principally
dedicate their time to the work in the milpa, the culture of bees and the breeding of
cattle. For sowing corn in the milpa, a traditional digging stick, called a choul, is
used and has undergone only one development since prehistoric times: the point,
originally hardened by the fire, has been replaced by a steel tip. Bullfights are the
central part of the feasts celebrated in honor of the Saint of the villages. Baseball,
introduced from the United States, seems to be the national sport of the Yucatec
Maya (Redfield and Villa Rojas, 1990 [orig. 1934]).
The fusion of the Mayan culture with aspects of the Christian faith are also present
in the incantations and prayers used for the treatment of patients or in the
numerous ceremonies. The modern ethnopharmacopoea of the Mayas consist
mostly of plants species, which originated on the Peninsula. But also introduced
ones are used, for instance Mentha spp. (Europe), Aloe spp. (Africa), Citrus fruits
(Asia) and are important medicinal plants.
25
Figure 2.4. Houses, new style and old style (poles, palm-leaf thatch)
Figure 2 5. House interior, kitchen
26
Figure 2.6. H-men (shaman) performing a
santiguar
Figure 2 7 Curandera (healer)
preparing herbal medicine
27
Medicinal Ethnobotany
Since about 1990, a tarred street connects the three villages with Peto and
Valladolid, two nearby towns. Electricity and thereupon TV changed the rhythm of
daily live (telephone since 1995). An increasing number of young people finish
secondary school in Chikindzonot and look for a job outside the villages,
particularly in the cities of Valladolid, Mérida and Cancun. Even though the
influence of the outside forces has been enormous, the Yucatec Maya still retain a
large number of ancient traditions.
2.1.5 Manuscripts on Maya medicinal plants
Medicinal plants played an important part in Ancient Maya culture. Evidence of this
is provided by old books and documents with medicinal characters. Two of them
are by indigenous writers, namely the Ritual de los Bacabes and Libros de Chilam
Balam, whereas the others are written by Spaniards (Gubler, 1997).
In the Ritual de los Bacabes (ritual of the spokesman/interpreter) incantations
and prayers for the treatment of disease are recorded. They reflect the concept
of Mayan illnesses and illustrate medicine to be closely associated with religion.
This unique Mayan work was written in the 18th century. The main author is
unknown, two pages were written by Joan Camul (Arzapâlo, 1987).
The Books of Chilam Balam are the sacred books of the Maya of Yucatan and
were named after their greatest prophet Chilam {Balam: Jaguar, priest).
Conventionally they are generally named after the towns in which they were
found. Three of the Books namely, Ixil, Kaua, and Nah include information on
medicinal therapy and mention several remedies against a variety of illnesses
(Gubler, 1997).
After the auto-da-fé in 1558, Diego de Landa wrote a history of the life of the
Maya of Yucatan. In the Relacion de las cosas de Yucatan (1565) and the
Relaciones de Yucatan (1580-1585), he explicitly mentioned that the Maya had
specialists for curing illnesses. He listed some medicinal plants and celebrated
28
Medicinal Ethnobotany
the beauty of the flora of Yucatan. His words indicate little confidence in
indigenous people (Diego de Landa, 1990 and 1992).
"There is in this land a great quantity of medicinal plants of various
properties, and if there were any person here who possessed a
knowledge of them, it would be most useful and effective. There is no
disease to which the native Indians do not apply the plants. But when
they are asked for an account of their properties, they can give none
other than that they are cold or hot, and that they are accustomed to
employ them to obtain the effect for which they apply them. However, as
a matter of fact, there are many of great virtue for every sort of illness
and as antidotes. On the other hand there are those which are
poisonous and deadly" (Roys, 1976).
In the Calepino de MotuI names of indigenous plants and some diseases as well
as some of the specialists who treat them are registered. The work was written
in the 16th century, when the Maya medicine was still largely unaffected by
Spanish influence (Roys, 1931). The book mentioned for instance: ac haban, a
herb with bad smell, which is used against cold and pechuguera (Gubler, 1997;
Arzâpalo, 1995).
El Libro del Judio is one of the most important studies of the medicinal plants of
Yucatan. The main part of the book gives a version, which is attributed to Juan
Francisco Mayoli, a roman physician who lived in Valladolid during the 18th
century and who used the pseudonym of Ricardo Ossado and the surname "El
Judio". The book contains a long list of indigenous uses and medicinal plants as
well as a few introduced ones. Furthermore it gives information on diseases.
The original work probably was written in the 16th or 17th century. Barrera and
Barrera Väsquez (1983) note that this is not just one book, but that several
copies and different versions of copies exist.
29
Medicinal Ethnobotany
An important source, which is not published, carries the title Libro de medicinas
muy seguro para curar varias dolencias con yerbas muy experimentadas y
provechosas de esta provincia de Yu[ca]than. It is a copy of an old manuscript
of 1751 and contains a list of medicinal plants and their prescriptions. Some of
the remedies incorporated European plants and elements. These documents
are preserved in the Bibliotheca Crescendo Carrillo y Ancona, Mérida (Gubler,
1997).
Another unpublished source, which can be found in the same library, is the
Relaciôn de las cosas y sus nombres de la provincia del YucalPeten written in
1710. Different themes are discussed and including two pages with a list of
indigenous medicinal plant (Gubler, 1997).
Two further books, Cuademo de Teabo and Cödice Perez, are mentioned as
containing medicinal texts (Gubler, 1997; Edmonson, 1986).
2.1.6 Medicinal ethnobotany of Yucatan
As part of a Mexican national evaluation, a list of medicinal plants was generated
based on publications, state inventories and student theses. This review includes
3,352 vascular plant species distributed in 1,214 genera and 166 families. Although
the vascular plant flora in Mexico has not been thoroughly explored, it is estimated
that it consists of at least 21,600 species. Hence, 15 % of the Mexican flora has
been employed for medical purposes. In order to produce catalogs of regional
medicinal plants, many Mexican institutions have compiled state inventories. The
states with highest percentage of locally documented medicinal plants are:
Quintana Roo (99 % of 373 species), Yucatan (60 % of 623 species), Veracruz (28
% of 548 species), Durango (26 % of 255 species), and Sonora (18 % of 548
species) (Bye etal., 1995; Argueta et al., 1994).
The flora of the Mexican part of the Yucatan Peninsula as mentioned before has
about 2,300 species of vascular plants and is therefore not very rich in comparison
with other regions in Mexico. Thus, Bye et al. (1995) points out that a greater
30
Medicinal Ethnobotany
degree of species richness does not necessarily indicate higher ethnobotanical
diversity. He suggests that the more phytochemically interesting plant-human
interaction is found in environmentally marginal or stressful areas than in species-
dense regions.
The botanical exploration of the Yucatan Peninsula began 1893. Several botanists
among them Gaumer, Millspaugh, Standley and Steyermark were dedicated to the
taxonomic investigation of the flora of Yucatan. Georg F. Gaumer, a practicing
physician who spent forty-five years in Yucatan, was keenly interested in medicinal
properties attributed to the plants by the indigenous people and in their Maya plant
names. Most of these voucher specimens are deposited in the Field Museum of
Natural History in Chicago (Standley, 1930).
In The Ethno-Botany of the Maya, Roys (1931) describes remedies, which are
essentially herbal but also contain animal and mineral materia medica. The texts
written in Maya and English were collected from several old books like El Judio,
and organized according to illnesses. The medicinal plants are identified based on
the works of Gaumer, Loesener, Millspaugh and Standley.
Mendieta and del Arno (1981) reviewed information about medicinal plants of
Yucatan and published this information in the Plantas Médicinales del Estado de
Yucatan. They also used secondary sources which are mentioned above. Others
are the Catalogos de Nombres Vulgares y Cientificos de Plantas Mexicanas
(Martinez, 1979) and Nomenclatura Etnobotanica Maya (Barrera et al., 1976). In
the latter book, the Mayan names of plants, their scientific identification, the origin
of the popular names and the etymological significance are discussed. Also here,
the information is based on secondary sources.
Alfonso Villa Rojas, born in Mérida, took charge of the Chan Kom school in 1927, a
village about 27 km from the study area of this thesis. He wrote several books and
publications about different aspects of the life of the Yucatec Maya. Together with
31
Medicinal Ethnobotany
Redfield, an anthropologist from the University of Chicago, he published the
ethnography Chan Kom - A Maya village, in which they included one chapter on
diseases, and their treatment as well as the meaning of nature (Redfield and Villa
Rojas, 1990 [orig. 1934].
One of the most important works on medicinal plants of the Yucatan, which
includes phytochemistry and toxicity, is the Atlas of Medicinal Plants of Middle
America - Bahamas to Yucatan of Morton (1981). The data here are also based on
secondary sources.
Until now no modern detailed ethnobotanical study on the Yucatec Maya has been
available. Thus, one of the main points of this study was to document the actual
use of medicinal plants of Yucatan and the scientifically identify the plant species.
The lack of pharmacological, toxicological and phytochemial studies of Mexican
plants provided a further point of the thesis. In the most recent work listing 2,049
medicinal plants of Mexico carried out by the Institute Nacional Indigenista (INI)
394 species were studied chemically, 280 species were studied chemically and
pharmacologically, 177 species were investigated chemically, pharmacologically
and toxicologically and from 69 species active principles were studied (Aguilar et
al., 1994; Argueta and Zolla, 1994).
32
Medicinal Ethnobotany
2.2 Methods in the field
Ethnobotanical data was collected during a total of 18 months, from February 1994
to June 1995 and from September 1996 to October 1996.
Qualitative and quantitative methods were used for the ethnobotanical evaluation
(Martin, 1996; Russell, 1988). Participant observations were carried out as one
of the methods used. The author lived with the Maya people and shared with them
many facets of their life: e.g. plant use in daily life, healing sessions, ceremonies,
cooking and marriages. Subjects like dreams, omen and witchcraft were discussed
in semi-structured or open-ended interviews. By means of questionnaires and
lists of questions and topics structured and semi-structured interviews were
held with 40 healers and midwives (Figure 2.8). They were interviewed
independently from each other. Only a few persons with non-specialist knowledge
concerning medicinal plants were interviewed. Excursions were made with the
specialists to their home gardens, the forests and along the borders of the paths in
the villages. The species were collected, pressed and dried in a field dryer
(wooden box of 70 x 120 x 80 cm, with a metal wire-lattice at the bottom and
underneath two electric bulbs to maintain a temperature of about 40 °C, Figure
2.9). Some interviews, prayers and songs during ceremonies were recorded on
tape with the permission of the healers. After becoming familiar with the medicinal
plants a portable herbarium was made and presented in some interviews. The
form of the interviews as well as the data obtained concerning medicinal plants
were documented and transferred to a database (FileMaker Pro ).
At the beginning of the field work, the ethnobotanical project was presented in a
meeting of the healers and midwives of Chikindzonot and Ekpedz. They were
asked to participate in the project. Initially, interviews with bilingual specialists were
carried out. Later on the interviews were held in Maya and translated to Spanish
with the help of a young woman of Ekpedz.
33
Figure 2.8. Questionnaire (in English and Spanish)
Popular name:
nombre popular
Scientific name:
Nombre cientifico
Number. Numero
Collection: Herbano
Picture- Foto
Informant, profession: Family.Flower:
Informador, profesion Familia
Floi
Seed, fruit:
Semilla, fruta
Uses: Collection' Leaf:
Usos Recolection Hoja
Identification Root:
Identificacion Raîz
Plant part used:
Partes utelisadas
Uses in other locations.
Utilizacion diferente que en el lugar de investigacion
Preparation:Preparacion
Description of the plant
Description
Doses, application (form,
time):
Habitat (vegetation type):
Dosis, aplicacion (forma,
tiempo) Habitat (tipo de veqetacion)
Effect: Observations:
Efecto Observaciones
Side effect, contra¬ Illustration of the plant:indication:
Efecto secundano, contra-
indicacion
Classification-
Clasificacîon Dibujode la planta
Description of the plant
(informant):
Importance of the plant (informant).
Description de la planta
(infomador) Importance de la planta (infomador)
34
Medicinal Ethnobotany
The author participated and supported the projects of the organization OMIMPY
(Organizaciön de Medicos Indigenas traditionales de la Peninsula de Yucatan)
during their meetings organized by the INI (Institute Nacional Indigenista). In order
to get a broader knowledge about the medical system of different regions of
Yucatan, the participating specialists in the meetings of this association were
interviewed and discussions were initiated. The ethnobotanical project was
presented at the secondary school of Chikindzonot and a small excursion was
carried out with the pupils. They were asked to collect one medicinal plant, which is
used in their homes, and to describe their medicinal effects. These species, dried,
pressed, labelled and covered with plastic, were deposited in the secondary school
at Chikindzonot. Other voucher specimens were deposited as described in
publications I, II and III.
Figure 2.9. Field dryer
The voucher specimens were identified with the help of botanists and specialists at
the CICY (Centro de Investigacion Cientifi'ca de Yucatan), Mérida and MEXU
(Herbario Nacional at the Universidad Nacional Autönoma de Mexico, D. F.). For
the plant evaluation of 48 species 100 - 200 g of plant material were collected. In
35
Medicinal Ethnobotany
the cases of the species which were of phytochemical interest, 1 - 2 kg of the plant
part used as part of a remedy were collected and dried in the shade.
The plant material was collected and exported with official permission of the
Secretana de Medio Ambiente, Recursos Naturales y Pesca - Instituto Nacional
de Ecologia, Mérida (22. April 1994 [No. 01245]; 12.April 1995 [No. 01105]; 21.
February 1996 [No. 686]) and Secretana de Agricultura, Ganaderfa y Desarrollo
Rural, Dirrecciôn General de Sanidad Vegetal, Mexico (Certificado Fitosanitario
Internacional, 12. May 1995 [No. 24075]; 23. October 1996 [No. 186]).
36
Medicinal Ethnobotany
2.3 Abbreviations mentioned in the plant list
Table 2.1. Plant use
No llnesses/conditions Enfermedades/condiciones
AT
1
counteract bites and stringsof venomous animals
snake bites
mordeduras y picaduras de
animales venenosas
mordedura de vibora
DER
1
dermatological conditions
inflammation
enfermedades
dermatologicasinflamacion
2 pimples granos
3 abscess absceso
4
5
inflammation of the throat
chickenpox
inflamacion de garganta
(mumps)viruella
6 measles sarampion
7 dermatomycosis hongos
8 pellagra pellagra
9 warts verrugas
10 psoriasis psoriasis
11 discoloration of the skin mal de pinto
12 scabies sarna
13 infection infecion
14 burning quemadura
EYE illnesses of the eyes enfermedades oculares
1 pain dolor
2 pimples granos
3 eye complaint problemas oftalmologicas
FEM women's medicine medicina para mujeres
1 spasm pasmo
2 problems of the vagina problemas vaginales
3 infertility infentilidad
4
5
pain of menstruation, disorder
of menstrual cyclechildbirth
dolor menstural, menstruacion
irregularinduccion del parto
6 "to induce" abortion induccion del aborto
7 inflammation of the vagina inflamacion vaginal
8 prevention of abortion prevencion del aborto
9 vomiting and fever duringconfinement
vomito and fiebre durante
alumbramiento (jobenal jolol)Gl gastrointestinal disorders afecciones gastrointestinales
1 diarrhea diarrea
2 dysentery dientena
3 mal de ojo mal de ojo
4 vomiting vomito
37
Medicinal Ethnobotany
5 spasm pasmo
6 constipation constipacion
7 bad air in the stomach mal aire en el estomago
8 parasites parasitas
9 stomachache dolor de estomago
10 mal viento mal viento
11 problems of the bile problemas de la billis
12 cirro cirro
OTH different uses otros usos
1 dandruff caspa
2 toothache dolor dental
3 pimples in the mouth herbes bucal
4 antidote antidoto
5 fractures of bone fracturas de huesos
6 hemorrhage hemorragia
7 vitamin deficiency deficiencia vitammica
8 earache doloren la oreja
9 ceremony ceremonia
10 splitting hair
Illnesses associated with
pelo con orquilla
PFE enfermedades asociadas con
pain and/or fever dolor y/o fiebre
1 rheumatism reumatismo
2 sweat during night, cold body sudacion nocturna, cuerpo fno
3 fever fiebre
4 headache dolor de cabeza
5 insomnia (lover's grief) insomnio (mal de amor)
6 trembling of babies tetano de bebes (dolor de
ombligo)7 invigorate the muscle tonifica los musculos
RES respiratory illnesses problemas respiratorias
1 catarrh catarro
2 bronchitis bronquitis
3 respiratory problems problemas respiratorias
4 cough tos
5 asthma asma
UR
1
2
3
4
urogenital problems
kidney trouble
diabetes II
anuresis
pain of the urogenital system
problemas urogenitales
problemas del nnon
diabetes II
anuresis
problemas urologicas
Medicinal Ethnobotany
Table 2.2. Plant part used
Abbreviation of Plant part used Parte usada de la planta
plant part used
ap aerial part hierba
ba bark corteza
bu bulb bulbo
fl flower flor
fr fruit fruta
gp green part hojas y tall os verdes
ju juice (watery) agua, jugo
la latex (milky) latex
Iv leaf hoja
pu pulp pulpa
re resin résina
rh rhizome rizoma
ro root rai'z
se seed semilla
st stalk tallo
tr trunk tronco
tu tuber tuberculo
wh whole plant planta entera
wo wood madera
so shoot retoho
Table 2.3. Application
Abbreviation for mode of
application
Mode of application Modo de aplicaciôn
con
lOG
nas
ora
pulrec
spi
vag
conjunctivallocal
nasal
oral
pulmonalrectal
spiritual
vaginal
conjuntivallocal
nasal
oral
pulmonal-rectal
espiritual
vaginal
39
Medicinal Ethnobotany
Table 2.4. Preparation
Abbreviation Mode of preparation Forma de preparaciön
M: combination with other plants combinaciön con otras plantas
bath bath baho
dec decoction decocto
drops drops gotas
empl emplastrum emplastruminf infusion infusion
lini liniment liniment
mac maceration rnaceracion
oint ointment pomada
pow powder polvo
soap soap jabon
syrup syrup jarabe
cer ceremony ceremonia
Table 2.5. Plant classification among the Mayas
Abbreviation Plant classification Clasificaciön de plantas
Tb bitter amargo
Ta astringent astringenteTe sweet dulce
Ts spicy picanteTi acid agrioTn no taste sin sabor
Sa aromatic, good smell buen olor
Ss strong smell olorfuerte, apestosoSb bad smell mal olor
Sn no smell sin olor
Sf little smell poco olor
Hh hot caliente
He cold frio
HI lukewarm tibio
Ho cool freso
2.4
Plant
list
Plantname(AANK#voucher)
Popularname
Use
Part
Applica-
Preparation
Ciassifica-
No.
of
used
tion
tion
resp.
ACANTHACEAE
Blechumbrownei(Kunth)(4
14)
ElytrariaimbricataPers.(310,449)
Ruellianudiflora(Engelm.&Gray)
Urb.
;
Syn.
:R.yucatanaTharp.
&Barkley
(115)
Ruelliasp.(528)
AGAVACEAE
Agaveamericana
L.(312)
Agave
maculata;
Syn.
:Manfredamaculata
(Reg
el)Rose
(197
)
Agave
aff.
fourcroydesLemaire(6
54)
YuccaelephantipesRegel
(416
)
ALLIACEAE
Alliumcepa
L.(6
41,642,643)
Alliumschoenoprasum
L.(4
63)
AMARANTHACEAE
Achyranthes
aff.
indica
Mill
.(5
37)
AMARYLLIDACEAE
(LILIACEAE)
Crinum
aff.americanum
(223
)
Crinumerubescens
Ait.
(408
)ANACARDIACEAE
AstroniumgraveolensJacq.(4
73)
Mangiferaindica
L.(4
09)
Spondiaspurpurea
L.(2
86)
ANNONACEAE
Annonamuricata
L.(630,631)
Annonapurpurea
Moc.&Sessé
(618
)
Annona
reticulata
L.(2
58)
AnnonasquamosaL
(261
)
Ak'abxiw
GI3PFE2
Ivfl
loc
bath
1
Kabalxa'an,Kambaxa'an
GI34
5,FEM1
2ap
ro
ora
loc
decbath
Hh
7
Kaba
lya'
axni
k,UR1
EYE1
ap
ora
loc
decM:bath
2
Sb
Maguey
FEM3
Ivora
M:
inf
Hh
Pets'k'im,
Pets'k'inil
PFE4
leloc
emp
He
Henequen
NI
IvTb
Tuk
PFE5
Iv
Ajo
FEM3
4PFE4
bu
ora
M:macdec
He
Cebollina
PFE4
bu
ora
mac
Bayche'
Gl6
ora
Sb
2 2 1 1 4 1
Pets'kini,
Pets'kinil
PFE4
Ivloc
empl
He
2
Xts'ulam
DER4
AT1
bu
loc
empi
1
-
RES3
Ivloc
1
Mango
FEM5PFE3
Ivloc
empl
Tn
1
Abal,Ciruela
DER5
6Iv
loc
M:
bath
iempl
SaTa
5
Guanâbana
RES4
Ivora
M:decsyrup
sweet
2
Poox
NI
fr1
Oop,Anona
DER1
PFE6
Ivloc
M:emp
ilbath
He
5
Saramuyo,Ts'almuy
GI4RES4
Ivora
loc
dec,bath
HeHhTa
9
ro
Plantname(AANK#voucher)
Popularname
Use
Part
Appl
ica-
Preparation
Classifica-
No.
of
used
tion
tion
resp.
Malmeadepressa
(Bai
ll.)
R.
E.
Fr.;
Syn.:
Guatteria
leio
phyl
la(F.D.Sm.)
Saff.Ex
Standi.(161
)
Sapranthuscampechianus
(Kunth)
Standley
(291,509)
APIACEAE(UMBELLIFERAE)
CoriandrumsativumL
(005
)
Pirnpinellaanisum
L.(2
55)
APOCYNACEAE
Catharanthusroseus
(L.)
G.Don
f.(2
71)
Echitesyucatanensis
Millsp.exSt
andl
ey(5
19)
Plumeriasp.(472,562,628)
Rauvolfiatetraphylla
L.(0
95)
Tabernaemontanaam
ygda
lifo
liaJacq.
(190229)
ThevetiagaumeriHemsl.
(324)
Urechites
andrieuxiiMuell.Ar
g.(4
41,446)
ARACEAE
Anthurium
schlechtendaliiKunth
ssp.
schlechtendalii(2
43)
Philodendronhederaceum
(Jacq.)Schott
(586,505)
Syngoniumpodophyllum
Schott(5
06)
ARISTOLOCHIACEAE
AristolochiaanguicidaJacq.(1
39)
AristolochiamaximaJacq.
(350
)
AristolochiapentandraJacq.(315)
Elemuy
UR1
2ro
Chuyuchajum,Sak-
UR1
NI
elemuy
Cilantro
GI4
Anisengranos
FEM5
GI1
Vicaria
FEM2
Sak-
vipe
rol,
Vipe
rol
rojo
DER1
AT1
Nikte'ch'om,
Flordemayo
DER7
Kambamuk
EYE2
DER8
NI
Uts'
upek
'DER1
9NI
Akits,
Cojönde
perro
DER1
AT1
Vipe
rolverde,Vi
pero
lAT
bejuco
Bobtun
FEM1
-
DER2
NI
-
NI
Wahk'oh-ak',Wahk'ohde
GI7FEM1
5
pato,Guaco
Wahk'oh
cast
illo
,Guaco
GI7FEM1
6
castillo
Wahk'oh
cast
illo
,Guaco
GH
2
castillo
se
se
Ivse
ro
re
Iv
ju-f
rre
re
re
ro
Iv
wh
ro
ro
ro
ora
ora
ora
ora
vag
loc
loc
loc
loc
loc
loc
ora
loc
ora
ora
ora
dec
dec
TbTaSs
Te
14
dec
Sa
11
powdec
SaTb
He
4freshbath
dec
M:decempl
Tb
2
drops
3
drops
Iv:TnTaSn
7
Sb
drops
Iv:TnSnSb
7
He
M:drops
2
freshempl
TbHe
5
M:dec
1
TnSn
2
TnSn
2
M:pow
inf
Sa
3
M:powdec
HhTbSa
1£
M:macdec
HhTsTb
3
Plantname(AANK#voucher)
Popularname
ASCLEPIADACEAE
(158)
Asclepias
curassavica
L.(0
70)
Cynanchum
schlechtendalii(Decne.)
Standi.&
Stey
erm.
(390
)Matelea
vindifloraWoodson;
Syn.
:Gonolobus
vindiflorusReom.
(022,
192,365,450)
Mateleayucatanensis(S
tand
i)Woodson
(345)
ASPHODELACEAE(LILIACEAE)
Aloevera
(L.)
Burm.f.;Syn..Aloe
barbadensis
Mill
.(065)
ASTERACEAE(COMPOSITAE)
AgeratumgaumeriRob.
(052
)
Artemisialudoviciana
Nutt.ssp.mexicana
(Willd.)
Keck(0
26)
Bidens
sp.(062
)Bidenssquarrosa
Less.
(121,
170,187,
404)
Galea
urti
cifo
liaMillsp.var.yucatanensis
Wussow,
Urb.&
Sullivan(1
35,153)
EupatonumodoratumL;Chromolaena
odorataR.M.King&
H.Rob.
(339
)Millenaquinqueflora
L.(3
69,413)
Montanoa
atri
phcifoliaC.Koch
(382)
Partheniumhysterophorus
L.(4
48)
Plucheasymphytifoha
(Mil
l.)Gillis
(009
,
109)
PorophyllumpunctatumBlake(152)
Tagetes
sp.(494
)
Tithoniadiversifoha(Hemsl
)Gray
(017
)
TrixisinulaCrantz(3
55)
VerbesinagiganteaJacq.(288)
Kalakts'u'um
Anal,
Ik'a
bal,
Polkuts
Xîum-ak'
Piin
-k'a
k',Kuyuch-ak',
Xp'okini,Emtsul
Ensul,Emtsul
Sâbila
,Petk'inki
Xpasmarxiw
Si'i
sim,
Artemisia
Sahun,Saksahun
Ya'axk'an-ak'
Xka'xikin,
Xikinkaax
Tok'aban
Xontolok
Xtankas-
ak',
Xuxtankas
Altamisa
Chalche',SantaMaria
Xuk'ii
Tempula
Arnica,Chaksu'um
Fluxionxiw,Xtankas-ak'
Chulkeeh
Use
Part
used
DER4
ro
OTH2
re
UR2
3re
Applica-
Preparation
Classifica-
No.
of
tion
tion
reso.
locora
empldrops
ora
drops
HI
ora
drops(d
ec)
OTH3
re
loc
drops
Hh
PFE6
loc
fresh
RES1
Ivora
M:dec
He
7
DER1
23
OTH1
loc
ointsoap
GI58
ap
ora
dec
Hh
6
GI48
Ivora
dec
inf
HhHcSaTb
26
PFE3
RES2
5Iv
ora
dec
2
GI3
Ivloc
M:
bath
8
DER2
10
Ivloc
M:bath
TB
10
Gl4
nas
inn
UR1
23
ro
ora
M"dec
TnSn
6
DER2
Ivloc
drops
3
GS10
ap
loc
bath
2
OTH4
FEM2
wh
loc
freshdec
2
FEM5
6Iv
ora
M:dec
inf
Hh
12
OTH
Ivloc
dec
Ss
4
FEM7
ap
loc
1
PFE
1Iv
loc
oint
5
PFE4
Ivro
loc
empl
2
RES2
Ivloc
empl
hot
1
ÊPlantname(AANK#voucher)
Popularname
Use
Part
Appl
ica-
Preparation
Classifica-
No.
of
_usecf
tion
tion
resp.
Wedelia
fertilisMcVaugh
(217
)
BASELLACEAE
AnrederavesicariaC.
F.Gaertner(116,
196)
BIGNONIACEAE
AmphilophiumpaniculatumKunth
var.
molle
(Schltdl.&Cham.)
(118
,172)
AmphilophiumpaniculatumKunth
var.
paniculatum
(144
)ArrabidaeafloribundaLoes.(2
07)
Ceratophytumtetragonolobum
(Jacq.)
Sprague&Sandw.
(180
)Crescentiacujete
Veil.(2
39)
Cydista
diversifoliaMiers.
(201,550)
Godmania
aesculifolia(Kunth)Standi.
(412
)Parmentieraaculeata(Kunth)Seem.;
Syn.:
P.edulisDC.
(107,096)
Parmentieramillspaughiana
(L.)
Williams
(135)
Pithecocteniumcrucigerum
(L.)
A.Ge
ntry
;Syn.:Pithecocteniumechinatum
Schum.
(002)
Stizophyllumri
pari
umSandw.
(156
)Tecoma
stansJuss.(0
04)
BIXACEAE
Bixaorellana
L.(2
33)
BOMBACACEAE
Ceiba
aesculifolia(Kunth)
Britton&Baker
(497
)CeibapentandraGaertn.
(456,614)
PachiraaquaticaAublet(4
37)
Sahun
Kaxichel
Sit'macho,
Petak'
Sosk
i-ak
',Xdunt'-ak'
Sak-ak'
Bakchiwoh
Luch.Jicara
Ek'k'ixil,
Soski-ak'
Xo'k'ab
Pepino
cat,
Kat
Katche'
Xache'ma'ax,Xache'xnuuk
AT1
DER2
DER1
OTH5
GI1
3
G12DER2
13
DER11
AT1
RES2
5
RES5
DER1
2NI
FEM5
7
UR1
23
UR1
IV
loc
1:bath
Xa'bach
RES2
5
K'anlol,
Tonadora
UR2
NI
Ki'wi',Achiote,K'uxub
GI9PFE3
PFE6
Pi'im
EYE1
DER10
Ya'axche'
PFE7
K'unche',K'
uych
e',Bonete
PFE3
Ivfr
ju-b
a
Iv ba
ora
loc
con
loc
loc
ora
pow
oint
drops
bath
drops
1
tu
loc
M:empl
HcTnSn
11
Ivloc
M:bath
Sa
4
Lv
loc
bath
2
Ivloc
M:bath
1
Ivpow
inf
ora
1
pu-fr
ora
dec
5
Ivf!
loc
M:empl
bath
SnSbTn
7
Ivfr
ora
M:dec
1
flfr
ro
ora
M:dec
SnTn
fr:Te
4
Ivora
M:dec
1
roseca
loc
M:pow
Sb
1
rofr
ora
dec
Te
1
ro
Ivfl
ora
dec
4
Plantname(AANK#voucher)
Popularname
Pseudobombax
elli
ptic
um(Kunth)
Dugand;
Syn.
:Bombax
elli
ptic
umKunth
(275)
BORAGINACEAE
CordiadodecandraA.DC.
(634,635,636)
Ehretia
tinifoli
aL.
(021
)
Amabola
bianco,
Sikl
ite,
Xk'uxche'
Cirricote,
Kop'te'
Beeb,
Roble,Xi'mche'
Hehotropiumangiospermum
Murr.(053,
131)
Toumefortia
volubilis
L.(126,165)
BRASSICACEAE(CRUCIFERAE)
Raphanus
sativus
L.(0
06)
BROMELIACEAE
Aechmea
bracteatavar.bracteataGnseb.
(167
)Bromeliakaratas
L.(3
73)
TillandsiabalbisianaSchuttes&Schultes
f.(625)
Tillandsiabrachycaulos
Schltdl.(4
96)
TillandsiaelongataScheudel
(479
)TillandsiaschiedeanaSteud.(6
26)
Tillandsiasp.(6
24)
BURSERACEAE
Burserasimaruba
Sarg
.(0
42)
CACTACEAE
Hylocereusundatus
(L)Britton&
Rose;
Syn.:CereusundatusHaw.
(427
)
Nopalea
cochenillifera
(L.)
Salm-Dyck
(313)
Selenicereusdonkelaarii
Britton&
Rose;
Syn.:CereusdonkelaniSalm-Dyck
(284)
Xnema'ax
Xulk
'ini
,Sal
Râbano
Ch'uk,Cinta
k'uk'
Ch'om,Ch'am,Pinuela
Ch'u
Ch'u
Ch'u
Ch'u
Ch'u
Chakah
Pitahaya
Nopal,Pak'am
Tsaran-ak'
en
Use
Part
Applica-
Preparation
Classifica-
No.
of
used
tion
tion
resp.
RES1
25
Ivba
ora
M:dec
Te
PFE3
RES25
ba
ora
dec
3
RES1
Ivba
locrec
M:dec
TeSa
15
PFE3
NI
ora
empl
GH268
ap
locrec
bathdec
HhTnSn
7
DER3
ap
loc
M.powempl
2
RES1
tu
ora
dec
2
FEM8
FEM5OTH6
FEM9
FEM4
FEM4
FEM9
FEM4
PFE2
3
GI2
RES2
UR4
OTH3
6
Iv Iv wh
wh
wh
wh
wh
Ivre
pu-l
v
Iv Iv
ora
Mmac
oraloc
decpow
loc
bath
ora
M.dec
ora
M:dec
loc
bath
ora
M:dec
loc
Hh
mac
bath
HeSs
ora
mac
HeTnSn
17
oraloc
macempl
Hh
3
ora
drops
5
èPlantname(AANK#voucher)
Popularname
CAESALPINIACEAE
Bauhiniadivancata
L.(0
07)
Bauhmia
herrerae
(Britton
&Rose)
Standi.
SSteyerm.
(124,148,442)
CaesalpmiagaumeriGreenman
(155,
344)
ChamaecristaglandulosaGreene
var.
flavicoma(Kunth)
H.
S.
Irwin&
Barneby;
Syn.:Cassia
g.(362)
Sennaatomana
(L.)
H.S.Irwin&Barneby;
Syn.
:Cassiaatomana
L.(2
73,337)
Senna
fruticosa
(Mil
l.)H.
S.
Irwin&
Barneby
(035
)Senna
obtusifolia
(L.)
H.S.Irwtn&Barneby
(361)
Sennaracemosa
(Mil
ler)
M.
Irwin&
Barneby
Senna
sp.(525)
Senna
uniflora
(P.
Mill
er)H.
Irwin&
Barneby
Senna
villosa
(Mil
l.)H.S.
Irwin&
Barneby;
Syn.:Cassia
villosa
Mill.(0
84)
Tamarmdus
indica
L.(2
69)
CAPPARACEAE
Cieomegynandra
L.(4
93)
CARICACEAE
Caricapapaya
L.(6
46)
CELASTRACEAE
Crossopetalumgaumeri
(Loes.)Lundell;
Syn.
:Mygindagaumeri
Loes.ex
Millsp.;Rhacoma
g.(0
38,128)
HippocrateaexcelsaKunth
(174
)
Mayvaca,Patadevaca
Kibix,
Mayvaca
rojo
,
Ts'ulubtok'
Kitamche'
Salatxiw,
Salat-ik'
Tu'ha'abin
K'anchik'in-ak'
Mehenbu'ul-xiw
Chululdzu
Xpahpul
Saalche',Boxsaal
Tamanndo
Papaya,
Put
Vipe
rolnegro
Sak-bo'ob.Xooknom
Use
Part
Appl
ica-
Preparation
Classifica-
No.
of
used
tion
tion
resp.
RES14UR12
ap
ora
Mdec
GI3
loc
Mbath
Gl3
Ivro
loc
Mbath
RES4
ora
Mdec
PFE4
Ivloc
pow
lini
EYE1
con
drops
Sa
10
7 4 3
PFE1
NI
Iv
GI3
Ivse
GH
PFE1
Iv
NI
Iv
NI
Iv
NI
Iv
DER2
Iv
GI1
ju-f
r
NI
Iv
AT1
re
AT1
ro
iv
GI1
loc
loc
loc
loc
ora
M-bath
M:
bath
bath
Sb
Tb
bath
decfresh
TbSb
Sf
TnSn
5 3 1 1 1 10
3
loc
drops
Tb
1
loc
ori
empl
fresh
TaTbHc
20
ora
M:dec
RES1
2Iv
ora
pow
inf
HhHc
Plantname(AANK#
voucher)
Popularname
Use
Part
Appl
ica-
Preparation
Classifica-
No.
of
used
tion
tion
resp.
•vl
CHENOPODIACEAE
ChenopodiumambrosioidesBertex
Steud.;Syn.:Teloxysambrosioides
(L.)
W.A.Weber
(028
)COMBRETACEAE
Terminaliacatappa
L.(3
26)
COMMELINACEAE
CommelinaelegansKunth
(093,106)
Rhoeo
discolorHance
(306
)CONVOLVULACEAE
Ipomoeabatatas
(L.)
Poir.(6
37)
IpomoeaheterodoxaStandi.&Steyerm.
(218
,248)
Turbinacorymbosa
(L.)
Raf.(0
46)
CRASSULACEAE
Kalanchoë
blossfeldiana
Poelln.(0
68)
Kalanchoë
intégraKuntze
(016
)Kalanchoëpinnata
Pers
.;Syn.:
BryophyllumpinnatumKurz(4
39)
CUCURBITACEAE
CayaponiaracemosaCogn.
(431
)
Cionosicyosexcisus(Griseb.)C.Jeffrey
(387
)Ibervilleamillspaughii
(Cogn.)C.
Jeff
rey;
Syn.:Co
ralo
carp
usmillspaughii
Cogn.
(094
)
Lagenaria
siceraria(M
olin
a)Standi.(3
83)
Luffaaegyptiaca
Mill.;
Syn.:
Luffa
cylindrica
Roem.
(149)
Momordicacharantia
L.(2
68)
Epazote,Lukumxiw
Almendra
G185
UR1
GI68
ro
Ivora
ora
dec
dec
Hh
after
25
cooking:
Sa
Ta
Uk'
ak'ah
ko'l
ebil
,EYE2
ju-fl
con
loc
3
Ya'axha'xiw
Chakts'am
DER78
11
Ivloc
empl
bath
3
Is,Camote
DER2
Ivloc
bath
Sb
1
Chiwohk'ax,Cancerxiw
DER12
PFE6
ro
loc
empl
bath
Hc
3
Xtabentun
PFE2
RES25
NI
Ivloc
M:bath
SbTb
3
FEM5
ro
ora
M:empi
Belladonna
DER1
Ivloc
empl
1
Belladonna
DER12OTH5
Ivloc
M:emp:
oint
Hc
11
Siempreviva
DER1
ivloc
1
Takeeyl
DER8
Ivloc
bath
-4
I
Kasam
DER8
Ivloc
empl
1
K'umkanul
DER1
PFE1
tu
loc
freshempl
SnSb
17
Lek
PFE1
pu-fr
loc
empl
1
Limpion
UR3
GI11
se
Ivora
dec
1
Morax
DER2
NI
Ivloc
bath
HITb
Sf
4
œPlantname(AANK#
voucher)
Popularname
SicydiumtamnifoliumCogn.
(036,336)
CYPERACEAE
Cyperus
articulatus
L.(0
15)
Sclerialithosperma
L.(1
75)
DIOSCOREACEAE
Dioscorea
spiculiflora
Hemsl.
(213
)
EBENACEAE
DiospyrosanisandraBlake(1
34)
Diospyroscuneata
Standi.
(431,424)
ERYTHROXYLACEAE
Erythroxyium
rotundifoliumLunan
(206,
379)
EUPHORBIACEAE
AcalyphaalopecuroidesJacq.(3
59)
Acalypha
sp.(4
32)
AcalyphaunibracteataMuell.Ar
g.(4
33,
507,581,593)
Astrocasiatremula(Griseb.)Webster
(455,504)
Cnidoscolus
aconitifoliusspp.
aconitifolius
(cultivated:
chayamansaMcVaugh)
(037)
CnidoscolussouzaeMcVaugh
(384
)Crotonchichenensis
Lundell(216,374)
Croton
humilis
L.(4
38)
Crotonlobatus
Forssk.;
Syn.:Jatropha
iobatusMuell.Ar
g.(1
38)
Croton
lundelli
iStandi.8040,
127,216,
374)
Chakmots-ak',Hoykep,
Saloli-ak',Cbikimu-ik'
Tupux
Xoknoon
Cancer-ak',
Wil-ak'
Xkakalche'
Sibi
l
Xik'iche'
Xmis
bil,
Mehenmis,Cola
degato
Sakpasmarxiw
Ch'ilibtux
il
P'ixt'onk'ax
Chaya,Chay
Chayademonte
Xebalam,Butsumukuy,
Xikinch'omak
lk-aban
Cruzoj
oxiw
Kokche'
Use
Part
Appl
ica-
Preparation
Classifica-
No.
of
used
tion
tion
resp.
EYE1
2AT1
Ivcon
loc
dropsempi
Hc
11
DER1
RES24
tu
ora
dec
Sa
4
RES24
ap
ora
M:pow
inf
1
AT1
Ivro
loc
dec
Hc
2
DER2
12
Ivloc
bath
Ss
13
DER1
NI
Ivloc
bath
Tb
4
DER2
Ivloc
bath
GI3
ap
loc
dec
TnSn
7
UR2
NI
ora
DER2
Ivloc
bath
1
Gl3
NI
Ivloc
bath
SnTb
7
NI
Ss
2
FEM1OTH67
re
Ivora
dec
Hh
2
PFE1
Ivloc
empl
1
DER2
12
Ivre
loc
bathemip
lHc
3
DER9
re
loc
drops
1
GI3
Ivloc
M:
bath
3
RES1
4Iv
ora
M:decinf
Hc
t
Plantname(AANK#voucher)
Popularname
CrotonperaeruginosusCroizat(1
32,205.
231,289)
Croton
reflexifoiiusKunth
(143
,377,399)
CrotonyucatanensisLundell(1
27)
Euphorbiaarmourrii
Millsp.(111,305)
Euphorbiahete
rophylla
Desf.(1
81,221)
Euphorbia
hirta
L.(3
64)
Euphorbia
aff.ocymoides
(112
)
EuphorbiaptercineuraA.Berger(0
34)
JatrophacurcasWall.(225)
JatrophagaumeriGreenman
(419)
ManihotesculentaCrantz(081)
Pedilanthusitzaeus
Millsp.(2
19)
Pedilanthusnodiflorus
Millsp.
(297
)
PhyllanthusacuminatusVahl(3
57)
Ek'balam,Xikm
burro,
Xikinch'omak
Pets'k'uts
Sakpokche',Ik-haab
Sakchakah,
Sibik'
Hobonk'ak'
Xanabmukuy
Kambalchakah
Much'kok
Sikl
ite'
Pomolche',Pinon
Yuca,Ts'nm
Ya'axhalalche'
Nabalche',Nahualte
Xulimil
4^CD
Phyllanthusglauscecens
Schltr.&Cham.
(228,261,526)
Phyllanthusmicrandrus
Muell.Ar
g.(0
88,
117,309,421)
Phyllanthussp
(202
)
Ricinuscommunis
L.(353)
Tragia
aff.
yucatanensis
Mill
.(459)
FLACOURTIACEAE
CaseanacorymbosaJacq.
(150,237)
SamydayucatanensisStandi.
(146,212)
XylosmaflexuosumHemsl.
(168
,209)
Zuelaniaguidoma(S
w.)
Bntt.&
Millsp.
(357,522,585)
ILLIACEAE
liliciumverumHook.
f.(255a)
LAMIACEAE(LABIATAE)
Hypt
issp.(4
75)
P'ix'ton-ak'
Pets'k'mi,
Kambaikiche'
P'ix'tonche'
Xk'ooch,
Higuerilla
P'op
'ox
Xi'mche'
Naranjache'
Puts'ukche',Xchaknif
Tamay,
Bot'ox
Anis
estrella
Xta'ulum,OreganoKaX
Use
Part
used
Appl
ica¬
tion
Preparation
Classifica¬
tion
N re
DER2OTH3
Ni
re
loc
decdrops
HcToTs
9
DER2OTH3
re
loc
drops
Hc
7
PFE3
Ivloc
inf
1
DER4
AT1
re
leloc
dropsempl
Hc
2
EYE1
re
con
drops
2
DER2
re
loc
drops
3
DER4
AT1
Ivloc
empl
1
RES1
24
reap
ora
decdrops
HhTb
8
DER2OTH3
re
loc
drops
TbTa
2
GI2
roju-s
tora
dec
TiTa
6
PFE2
DER212
Ivloc
decbath
Sb
2
AT1
wh
locora
empldec
Hc
1
OTH2
3re
ora
drops
Hc
2
DER1
2NI
Ivloc
decempl
Hh
6
bath
UR1
NI
Ivba
ora
dec
SsTb
4
DER1
OTH26
Ivap
locora
empldec
9
DER1
Ivloc
empl
1
GI6PFE34
Ivse
ora
loc
decempl
4
PFE1
Ivloc
empl
lini
2
PFE3
DER12
NI
Ivro
loc
bath
TbSa
6
UR23
ro
ora
dec
4
AT1
DER3
Ivloc
empl
2
FEM1
ro
ora
syrupdec
Hh
5
GI7
frora
M:dec
1
GI3
Ivloc
M:
infbath
Sb
1
en
Plantname(AANK#voucher)
Popularname
Use
Part
Appl
ica¬
Preparation
Classifica¬
No.
of
used
tion
tion
resp.
Leonotisnepetaefolia
(L.)
R.
Br.(4
05)
-PFE4
Ivloc
mac
1
Mentha
aff.
arvensis
L.(029)
Hierbabuena
GI459
ap
ro
ora
M:dec
SaTe
13
Mentha
aff.
citrataEhrh.(0
30)
Toronjil,Balsamo
GI45OTH2
Ivro
ora
M.dec
Sa
14
Mentha
aff.
pipe
rita
L.(0
31,045,054)
Menta,Balsamoxiw
GI49PFE4
ap
Ivora
M:mac
inf
dec
SaTaHh
23
Ocimum
basilicum
L.(0
71)
Albahaca
PFE4OTH2
ap
locspi
M:decdrops
16
EYE1
se
con
fresh
Ocimummicranthum
Willd.(033,147)
Xkakaltun,Albahacade
GI2
ju-l
vap
ora
Mdrops
HcSaTa
13
monte
DER2
ap
loc
Mbath
oint
Origanumvulgare
L.(6
38)
Oregano
FEM6
Ivora
dec
1
Plectranthusamboinicus(Lour
)Spreng;
Oregano
cast
illo
,Oregano
OTH8
ju-l
vloc
drops
Hc
8
Syn..Coleusamboinicus
Lour.(0
13)
chino
Rosmarinus
officinalis
L.(589)
Romero
FEM5
Ivora
Mdec
Hh
1
SalviacoccineaJuss.exMurray
(110
)Pezuhade
caballo,Unas
de
caballo,Chaktsits
GI2
NI
ro
ora
mac
Ta
3
SalviamicranthaDesf.(0
25)
Xiax-k'ax,
Chi-k'ak',
DER12
13
ap
loc
Mbath
SaTeHcTb
14
Contrahierba
RES1
GI10
Ni
ora
dec
Satureja
brownei(S
w.)
Briq
.;Sy
n.:
Poieo
PFE4
Ivora
loc
M:decmac
SaHe
15
MicromenabrowneiBenth.
(044
)Scutellaria
aff.gaumenLeonard
(338
)Orozuz,Balsamoxiw
GH
4ap
ora
Mdec
1
LAURACEAE
Perseaamencana
Mill
.(4
53)
On,Aguacate
UR1
2RES4
Ivora
M-decsyrup
TeTa
4
LOASACEAE
Gronoviascandens
L.(3
58)
Laalmuch
PFE1
Ivloc
empl
1
LOGANIACEAE
Spigelia
sp.(5
35)
Lombnzero
GI8
wh
ora
dec
1
MALPIGHIACEAE
BunchosiaswartzianaGnseb.
(142,
173,
Sipche'
GI3PFE4
NI
wh
spi
purifie
6
552)
bath
Byrsonima
crassifoliaKunth
(426
)Chi1,Nance
GI1
DER2
6Iv
ba
ora
Mdecbath
5
MALVACEAE
(024)
Taman-ka'an
PFE4
GI10
se
locora
powempl
2
AbelmoschusesculentusMoench;
Syn.:
Caféchino
AT1
se
loc
powempl
1
Hibiscusesculentus
L.(0
19)
Plantname(AANK#
voucher)
Popularname
Abutilonpermolle
Sweet(0
91)
Gaya
calyptrata
Kunth
(356
)
Gossypium
hirsutumCav.
(086
)
Gossypium
sp.(632,647)
Hibiscusrosa-sinensis
L.(0
67)
Hibiscustubiflorus(Moç&Sessé)DC.
(120,474)
MalvaviscusarboreusCav.
var.arboreus
(140,189,597)
SidaacutaBurm.
f.(3
66)
Sida
aff.
rhombifolia
L.(4
42,588)
MARANTACEAE
Marantaarundmacea
L.(4
64,558)
MARTYNIACEAE
Martyniaannua
L.(4
95)
MELIACEAE
Cedrelaodorata
L.(3
01)
TnchiliaarboreaC.DC.
(503
)Tnchilia
hirta
L.(5
14)
MENISPERMACEAE
Cissampelos
pareira
L.(0
08)
MIMOSACEAE
Acaciaangustissima
Mill
.(378)
Acacia
collinsn
Saff.(0
39)
Sakmisbil,Sakpetmis
Xpup
ul-i
k'
Taman,Algodôn
gris
Chuy-taman
Tulipan
Bisil,Xcampana
ka'ax
Bisi
l-ch
e',
Bisil,
Holol
Chichibe
Pasmarxiw
Chaak
Unade
gato
,Carnavalia,
Sarsapanlla
Cedro
Chobenche'
Chobenche'
Peteltun,Orejade
ratön
Waxim
Subinche'
Acaciasp.(432
,640)
Desmanthus
aff.
depressusHumb.&
Bonpl.ex
Willd.(1
84,287,298)
EnterolobiumcyclocarpumGnseb.
(018
)
Leucaenaleucocephala
(L.)
deWit
(481)
Lysiloma
latisi
liqu
aBenth.(3
80)
Mimosabahamensis
Benth.(3
42)
Ch'imay
Sala
t-ik
',Sibik'xiw,
Sib-
ik",
Sik'ink'ax
Pich
Waxim
Tsalam
Sak-katsim,Katsim
Use
Part
Appl
ica¬
Preparation
Classifica-
No.
of
used
tion
tion
resp.
OTH3
NI
Ivloc
fresh
TnSn
Sb
8
GI3DER2
Ivro
ora
loc
decbath
2
RES
young
frora
M:syrupdec
Te
2
RES1
Ivora
RES4
5Iv
ora
M:dec
2
GI3
Ivfl
loc
M:bath
2
GI2
Nl
ro
ora
macdec
HcSnTn
13
GI3OTH3
re
loc
decbath
2
GI510
9P
ora
Mdec
Hh
8
GI23
rh
ora
ju
2
UR3
trora
dec
1
OTH6
Ivnas
fresh
Ss
6
NI
IvTb
Sn
3
NI
IvTnSn
1
GI3
loc
M-
oath
10
DER2
Ivloc
bath
HITb
1
AT1
DER12
Ivloc
powempl
dec
HhHc
4
GI6DER2
ro
Ivora
loc
dec
2
EYE1
ivcon
drops
4
GI310
NI
Ivloc
Mbath
3
DER1
ivloc
dec
Hc
2
OTH2
ba
loc
empldec
Hh
1
RES4
roba
ora
dec
3
r5Plantname(AANK#voucher)
Popularname
PitheceliobiumalbicansBenth
;Sy
n.:
Havardiaalbicans
Britton&Rose
(166
)
Pitheceliobiumdulce(Roxb.)Benth.(4
99)
Pitheceliobiummangense.;
Syn.
:Chloro-
leuconm.
Britton&Rose(163)
MORACEAE
Brosimum
alicastrumSw
(092
)
Cecropia
obtusifoliaBertol.(0
41)
Chlorophoratinctona(L
)Gaud.(515)
DorsteniacontrajervaL
(330
)
Ficus
cotinifoliaKunth
(020
)
MUSACEAE
Musasp
(539
,541,542,543.544)
MYRSINACEAE
Ardisia
aff.
escallonioidesSchltr.&Cham.
(436)
MYRTACEAE
Calyptranthes
millspaughii
Urban.
(517
)
Eucalyptussp
(613,629)
Eugenia
aff.
capuli
(Schlecht&Cham
)
Berg
(479)
Eugenia
buxifolia(Sw
)Willd.(1
33)
Pimenta
dioica
(L)Merr.;Sy
n..Pimenta
officinalisLmdl.
(023
)Psidiumguajava
L.(2
35)
Chukum
Ts'iuche'
Xiax-ek'
Ox,Ramon
Xk'oochle',Guarumo
Morax
Xkambalhaw
Kopo
',Alamo
Plâtano,Ha'as,Platano
manzano
Xook'num
Eucalyptus
Chaknii
Xhilnich',
Sakloobche'
Nohochpol,Pimientade
Tabasco
Pich
i',Guajaba
Psidiumsartonanum
(Ber
g)Nied
(211
)
NYCTAGINACEAE
Boerhaviasp.
(471
)
Neeapsychotnoides
F.D.Sm.
(274,512)
Pisoniaacuieata
L.(154,249,407)
OLACACEAE
Ximeniaamencana
L.(4
18)
Pichiche'
Chakle'
Xtatsim
Beeb,Unadegato
Nabche',Tsu'tsup
Use
Part
Appl
ica-
Preparation
Classifica-
No.
of
used
tion
tion
resp.
DER1
79
ba
loc
bath
HhHc
7
NI
IvSnTb
1
AT1
DER3
Ivloc
M:powempl
1
RES4
5re
ora
drops
9
UR
Ivora
Mdec
5
NI
IvTa
1
GI7FEM3
rh
ora
pow
infdec
HhSaTb
27
RES4
NI
re
ora
drops
4
GI1
ora
Mfreshdec
Hh
RES45
loc
empl
NI
IvSaTi
2
RES45
Ivora
Mpow
inf
2
DER12
EYE3
ba
Ivloc
1
DER2
Ivloc
bath
Tb
3
FEM2
34
frora
M.dec
Hh
14
GI4
Ivloc
GH
frora
dec
fresh
Iv:HhTaTb
34
DER2
6Iv
loc
bath
Safr:Hc
DER2
Ivloc
bath
TaSa
HI
12
DER2
12
loc
Mbath
H
DER2
13
NI
frIv
loc
bath
2
FEM5
so
ora
dec
Tb
8
GI1
2ro
ora
Mdecmac
Plantname(AANK#voucher)
Popularname
ORCHIDACEAE
CatasetumintegerrimumHook.
(188,302)
Ch'itku'uk
Encyclia
aff.
belizensis(Reichb.
f.)Sc
hltn
.;Xkananikte'
(425
)Oncidiumascendens
Lind
ley(244)
Puts'ubche',Bac
chivo:
Puts'maskab
Oncidiumcarthagenense(J
acq.
)Swartz
U'tsumpek
(245)
Spirantessp.(2
41)
Chiwohk'aak'
OXALIDACEAE
Oxalis
lati
foliaKunth(352,612)
Elel
PALMAE
Cocos
nucifera
L.(650)
Coco
Sabal
sp.(633,648,649)
Guano,
Ka'nal-xa'an
PAPAVERACEAE
Argemonemexicana
L.(026)
Carmesanta
PAPILIONACEAE
(465)
Chaksaal
(345
)Saklooche'
Abrus
precator
ius
L.(001)
Oxo
Aeschynomene
fascicularisCham.&
Salat-ik'
Schlt.(0
63)
Canavaliaensiformis
(L.)
DC.
(513
)Canavalia
Centrosema
sagittatum(Humb.&
Bonp
l.Buy-ak'
ex
Willd.)Malme;
Syn.
:Glycine
sagi
ttat
aHumb.&
Bonpl.ex
Willd.
(128
,137)
Daleacarthagenensis
var.barbata
Azüfrexi
w,Suyk'ak'
(Oer
st.)
Barneby
(125
,151,178)
Desmodium
aff.canumSchinz&
Thell.
Pak'umpak'
(367
)Desmodium
sp.(523)
-
mDiphysacarthagenensisJacq.
(435
)co
Susup,Ts'us'uk
Use
Part
Appl
ica-
Preparation
Classifica-
No.
of
used
tion
tion
resp.
DER1
13OTH5
so
loc
M.empi
Hc
7
DER1
bu
loc
1
DER9
Ivloc
empl
1
FEM3
ora
M:dec
1
DER2
7ro
loc
pow
Tb
2
PFE3DER2
Ivora
dec
Ta
2
FEM8
ju-f
rora
fresh
1
FEM349
NI
so
ora
dec
Hh
3
UR3
PFE3
RES2
Ivse
ora
dec
DER2
Ivloc
bath
Hh
1
DER2
Ivloc
dec
1
GI3
Ivloc
bath
Hc
12
DER1
Ivloc
M.powempl
1
NI
IvTbSn
EYE1
AT1
ro
Ivcon
loc
2
DER2
Iv
DER1
Iv
Nl
Iv
ATI
G13
Iv
loc
loc
dropspow
4
empl
1
SnTn
1
M:empl
bath
Te
5
$5Plantname(AANK#voucher)
Popularname
ErythnnastandleyanaKrukoff(3
79)
Chakmolonche'
Indigofera
jamaicensisSpreng.
(360
)Xoxo-ak'
Indigofera
suffruticosa
Mill
.(2
08)
Sujuxiw
Lonchocarpuspunctatusssp.
longistylus
Balche'
(351)
LonchocarpusxuulLundell(1
77)
Xuul
Mucunaprunens
(L.)DC;Syn/
Stizo-
Xpica
lobium
prunens
(L.)
Medic.
(518,549)
NissohafructicosaJacq.(3
43)
Xk'ant'uul
Pachyrnzuserosus
(L.)
Urban
var.
Kup,Jicama
palmatilobus(D
.C.)
Clausen
(346
)Piscidiapiscipul
a(L
.)Sarg.
(123,320)
Ha'abm
PASSIFLORACEAE
PassifloraconaceaeJuss.
(246
)
Passiflorafoetida
L.(4
52)
PHYTOLACCACEAE
PhytolaccaicosandraSims
(388)
Rivinahumilis
L.(089,308)
PIPERACEAE
PiperamalagoL
(098
,105)
POACEAE(GRAMINEAE)
Cymbopogon
citratus(Nees)
Stap
f;Syn.:
Andropogon
citratusHortexDC.
(061
)Lasiacis
ruscifoliaHitchc.&Chase
var.
ruscifoiia(3
33)
Zeamays
L,(363
,575)
POLYGONACEAE
AntigononleptopusHook.&
Arn.
;Syn.:A.
cordatum
Mart.&
Gal
(237
)CoccolobaspicataLundell(5
10)
Coccolobauvifera
L.(5
45)
Xik'sots'
Xpoch
T'eikox
Colario,
Ikiche*
Xpeheche'
Zacatede
limon
Sut
Maïs,Cabellode
elote
Atol
e,Pinole
SanPedro
Bob
Uvademar
Use
Part
Appl
ica-
Preparation
Classifica-
No.
of
tion
resp.
EYE2
se
con
pow
3
RES5
UR3
ora
M:dec
Gl3
Ivloc
bath
1
GI2
Ivloc
bath
1
RES5
PFE3
ba
ora
dec
HhTnSs
3
OTH9
NI
cer
PFE134
ro
loc
bath
Jim
5
GI6
frora
mac
2
AT1
ro
Ivloc
empl
1
RES1
24
frro
ora
freshdec
TeHc
3
GI2
RES4
OTH8
Ivlocdrops
1
DER12
frlocfresh
2
DER6
froralocdecbathTe
9
AT1
DER8
rolv
loc
empl
oint
HcSb
Tb3
Ivora
mac
syrup
lb
IvlocdropsfrlocfreshfroralocdecbathTe
ro
Iv
loc
empl
oint
HcSb
IvlocfreshsoapSsIvora
dec
HhTeIvlocorafreshempl
TnSn
froraM:dec
OTH10
NI
IvlocfreshsoapSs
10
RES1
4GI9
Ivora
dec
HhTe
12
DER9
NI
ivlocorafreshempl
TnSn5
UR1
2GI14
froraM:dec
7
RES4
5rofl
ora
dec
1
OTH3
DER2
NI
re
Ivloc
dnoos
TnSn
3
UR1
Ivora
dec
1
Plantname(AANK#voucher)
Popularname
Gymnopodium
floribundumRolfe(3
72)
Ts'i
ts'i
lche
'
NeomillspaughiaemarginataS.
F.Blake
Sakitsa',Xtastabin
(203)
POLYPODIACEAE
Microgramma
nitida
(J.Sm.)
A.ReedSm.
Tipte'-ak'
(183,303)
PRIMULACEAE
Samolus
ebracteatusKunth
(032
)Tsunya'hi
PUNICACEAE
Punicagranatum
L.(2
72)
Granada
RANUNCULACEAE
Clematis
dioica
L.(520)
Xmexmexib
en
cn
RHAMNACEAE
Colubrinagreg
giS.Watson
var.
yucatanensis(5
00)
Gouania
lupuloidesUrban
(376,417)
KrugiodendronferreumUrban
(498
,571,
578,623)
ROSACEAE
Rosachinenesis
L.(1
30)
RUBIACEAE
Borreriasp.
(470
)Borreria
vert
icil
lata
G.Meyer
(276,608)
ChiococcaalbaHitchc;
Syn.
:C.
racemosaL.
(215,240)
Coffeaarabica
L.(2
70)
HameliapatensJacq.
(199
)MorindayucatanensisGreenman
(113
)
PsychotriamicrodonUrban
(329
)
PsychotriapubescensSw.
(400
)Randia
longil
obaHemsl.
(415
)
Pujuche'
X-om-ak'
Chintok'
Rosa
ychina
Haway
Haway,Haway-k'ak'
Chimes-kas,
Xiax-al'
Café
Ele'kabi,K'anan
Piha
ak',
Pihakam
Xbakalik'
Tschul-keeh
K'ax
Use
Part
Appl
ica-
Prep
arat
ion
Classifica¬
No.
of
used
tion
tion
resp.
RES1
5flro
ora
dec
Ss
3
DER11
AT1
NI
RES1
so
loc
bath
SnTb
3
GI12
wh
ora
dec
HcSnTb
17
DER1
4OTH5
Ivloc
M:empl
GI1
frora
M:fresh
DER2
Ivloc
M:
bath
TaTi
12
12
DER2
Ivloc
1
NI
IvSbTb
4
OTH3
NI
ro
ora
mac
HcTbSn
5
UR1
ba
ora
dec
4
OTH2
wo
loc
empl
RES1
45
Ivora
dec
8
DER12
ap
loc
M:
bath
1
DER2
12
ap
loc
bath
Hh
10
DER2
Ivro
loc
pow
bath
Tb
9
FEM5
se
ora
inf
Tb
1
DER1
2Iv
loc
emplbath
Hh
5
DER9AT1
frlv
loc
empl
HcSb
9
PFE4
Ivloc
M:empl
bath
1
PFE34
Ivloc
mpi
bath
3
OTH9
frsp
ifresh
3
en
CD
Plantname(AANK#
voucher)
Popularname
Use
Part
Appl
ica¬
Preparation
Classifica¬
No.
of
used
tion
tion
resp.
Yuy,Sihun
G19RES1
PFE12
NI
Ivro
loc
decbath
9
Limön
pals,Limönagna
RES4
5ju
ora
drops
HcSaTbTi
27
GI5
Ivdec
dec
Pak'aal,Naranja
agri
aGI47
Ivora
dec
HhSa
18
Lima
GI3
Ivloc
dec
1
Mandarine
Gl7
Ivora
M:dec
1
Naranjadulce,China
GI7
Ivfr
ora
M:decdropsSaTb
13
Cajera
GI1
ro
ora
powmac
Hh
1
Limonaria
RES2
5fl
loc
pow
oint
4
Tamkasche',
Siische'
GI910RES5
ro
ora
linidec
Hh
11
Ruda
PFE4GI1
37
ap
Ivloc
M:macdec
Hc
24
Sinanche',Matade
PFE24
GI10
ro
Ivju
loc
pow
lini
HISb
16
escorpiôn
-
DER2
NI
Ivloc
SaSnTn
3
Sihum
NI
IvTbTnSn
5
Chi'keeh,Caimito
silvestre
GI127
ro
ora
M.powmac
2
Ya',
Zapote,Chicle-zapote
G12
ba
ora
M:dec
HcTa
Ti
32
Zapote
bianco,Sakya'
G11
frora
dec
3
RUTACEAE
CasimiroatetrameriaMillsp.(049)
Citrusaurantiifolia(Christm.)
Swingle
(257
)Citrusaurantium
L.(2
36,253)
Citruslemon
(L.)
Burm.
f.(6
51)
Citrus
reticulataBlanco(2
52)
Citrussinensis
(L.)Osbeck
(247,251)
Citrussp.(236
)
Murrayapaniculata
(L.)
Jack.
(099
)
PilocarpusracemosusVahl(4
54)
Rutachalepensis
L.(0
55)
ZanthoxylumcaribaeumLam.
(368
)
SAPINDACEAE
Allophyloscominia(L)Swartz(389,511)
Sapindussaponaria
L.(5
08)
SAPOTACEAE
ChrysophyllummexicanumBrandegee
(386
)Manilkarazapota
(L.)
Royen;
Syn.:Achras
zapota
L.;Sapotaachras
Mill
.(2
34)
Pouteriacampechiana
(Kunth)Baehni;
Syn.:LucumacampechianaKunth
(108)
Pouteriasapota
H.
E.MooreandSteam;
Syn.:Calocarpummammosum
Pierre
(540
,547)
Pouteriauniloculars(Donn.Smith)Baehni
(619
)SCROPHULARIACEAE
Capraria
biflora
L.(0
97)
Mamey
Zapote
amarillo
Claudiosa,Sak
clav
iosa
,
Chokwil-xiw
GI1
NI
rose
se
ora
dec
GI5OTH8
NI
loc
drops
Hc
Plantname(AANK#
voucher)
Popularname
RusseliasarmentosaJacq.(1
60,340)
SELAGINELLACEAE
Sela
gine
llalo
ngis
pica
taUnderw.
(214
)
SIMAROUBACEAE
AlvaradoaamorphoidesLiebm.
(136
)
SOLANACEAE
CapsicumchinenseJacq.
(265
)
Cestrumnoctumum
L.(0
50)
Datura
aff.
inoxia
Miller(4
58)
Nicotianatabacum
L.(0
82)
Physaliscinerascens(Dunal)
Hitch.(5
02)
Siik
'xiw
,Oxletk'ax
Mooch-tut,
Flordepiedra
Belsinikche',Palode
hormigas
Habanero
Juandenoche
Chaniko,Chamisa
K'uts,
Tabaco
Xpurusi
Solanum
aff.
armentalis
J.Gentry(4
47)
Xsikli-much
en
-vl
Solanumcandidum
Lind
ley(s
pec,
related
toS.hirtumVahl.)
(440
)SolanumerianthumG.Don
f.(334,530)
Solanum
hirtumVahl.(0
90)
Solanumnigrum
L.(2
67,420)
SolanumrudepannumDunal(s
pec.
relatedtoS.torvum)(4
69)
Solanum
sp.(5
83)
Solanum
sp.(5
27)
STERCULIACEAE
Guazuma
ulmifoliaLam.
(250
)
HelicteresbaruensisJacq.
(176
,553)
THEOPHRASTACEAE
Jacquiniaaurantiaca
Ait.
(371)
TILIACEAE
Lueheaspeciosa
Willd.(3
47)
TriumfettasemitrilobaJacq.
(230
)
Triumfetta
aff.
ulmifolia(2
48)
Papera
Xpahhux,Ukuch
kax
Putbalam
Hierbamorax
Xsikli-much
Tomate,
p'ak
'
Xpuh-hi
Pixo
y,Nohoch-pixoy
Tsutup,Suput
Sink'inche'
K'askat
Mul-och
Kambapixoy
Use
Part
Appl
ica¬
Preparation
Classifica¬
No.
of
used
tion
tion
resp.
AT1
DER2
ap
loc
empl
bath
6
UR1
ap
ora
powdec
2
DER2
710
NI
Ivloc
bath
HhSnTe
9
PFE6
frloc
powemp
2
DER1
PFE6
Ivro
loc
M:bathempl
Hc
8
DER2
Ivloc
ointempl
3
DER2
12
Ivloc
M:dec
lini
Hc
5
NI
IvTbSb
2
DER2
Ivloc
M:dec
oint
empl
2
DER2
frloc
empl
2
DER13
NI
Ivloc
emlp
2
AT1
frloc
empl
1
DER113
Ivloc
empl
Hc
5
DER2
Ivloc
bath
1
DER14
ju-l
vloc
drops
1
NI
IvSb
1
FEM5
ba
ora
dec
HhSa
12
OTH11
frsp
ifresh
10
OTH9
spi
amu
Hh
AT1
DER2
Ivloc
empl
2
GI2FEM5
ro
ora
decmac
HhHcSnTn
26
FEM4
5Ivba
ora
dec
HhSa
8
öSPlantname(AANK#voucher)
Popularname
Use
Part
Appl
ica-
Preparation
Classifica-
No.
of
used
tion
tion
resp.
TURNERACEAE
Turneradiffusa(Willd.ex
Schu
lt.)
(193
)
URTICACEAE
UreracaracasanaGaud,ex
Griseb.(3
25)
VERBENACEAE
Callicarpa
acuminataRoxb.
(169
)
CornutiapyramidataAiton(0
48)
Durantarepens
L.(222)
Lantanacamara
L.(2
82)
Lippia
albaN.
E.
Br.Ex
Britton&Wilson
(011)
Lippia
dulcisTrev.;
Syn.
:Phyla
scaberrima
(Juss.
ex
Pers.)
Moldenke;
ZapaniascaberrimaJussex
Pers.
(059)
Lipp
iaaf
f.graveolensKunth
(014
,554,
611)
Lipp
iastoechadifoliaKunth
(010
)
Petrea
volubilis
Veil.(003,119)
PrivalappulaceaePers.(4
30)
StachytarphetajamaicensisGardner(027,
521)
Stachytarpheta
sp.(5
36)
VitexgaumeriGreenman
(047
)
VIOLACEAE
Hybanthusthiemei
(F.Donn.Sm.)Morton;
Syn.
:Indiumthiemei
F.Donn.Sm.
(266)
Hybanthusyucatanensis
Millsp.(533,534)
VITACEAE
Cissus
trif
oliata
Lour.(0
43)
Oregano
k'ax
,Oreganode
RES4
Iv
monte
Laal,Or
tiga
,Pica--pica
GI9
PFE1
4Iv
Xpuk
'in,
Puk'im
GH
2so
Xolte'xnuk
RES1
PFE1
3Iv
K'anpokolche'
DER9
NI
fr
Tédemonte
GI4
Iv
Tédelimön
Orozuz
GI4
PFE3GI4
Iv Iv
Oregano
FEM6DER8GI4
ora
loc
ora
loc
loc
ora
ora
ora
ora
dec
Sa
dropsmac
Hh
dec
7
mac
bath
bath
drops
inf
HcTbSa
HcHc
SnTa
HhTn
12
9 2 1
mac
SaHc
10
dec
2
Tédechina
GI4
Ivora
dec
HcHhSa
29
Yochop'tsimin,Comidade
GI3
Ivfl
loc
bath
3
caballo
Xpak'umpak'
AT1
Gi7
Ivloc
empl
bath
2
Verbena,
Iben-xiw
PFE1FEM4
Ivloc
M:
lini
5
Malva,Verbenaxiw
NI
IvSs
1
Ya'axnik
PFE2
Ivloc
bath
Sa
1
Xpluxionxiw
OTH2
PFE4
ap
locora
dec
Hc
5
Sakbakelkam
AT1
NI
ro
loc
empl
TnSn
1
Cruzoj
oxiw
GI3
Ivloc
bath
14
Plantname(AANK#voucher)
Popularname
Vitistr
ifol
iataHumb.&
Bonp
l.exRoemer
Tabkanih
&Schultes(5
24,531)
WINTERACEAE
Drimys
winteriForster&
Forster
f.(2
56)
Canela
ycuyo
ZINGIBERACEAE
Zingiber
officinaleRoscoe
(370
)Em
ojib
le,(J
engi
bre)
(566)
Katku'ut
(574)
Muela
(476)
Oxo
k'ax
(157)
Ts'u
ts'u
pche'
(570)
Tusik'
(563)
Xiek'
in
enCD
Use
Part
Appl
ica-
Preparation
Classifica-
No.
of
used
tion
tion
resp.
NI
ivTnSn
G!
ba
ora
po'w
inf
HhSaTb
1
GI7
rh
ora
M:powmac
HhSaSs
8
UR2
ro
ora
M:dec
1
DER2
Ivloc
M:bath
1
GI3
frloc
M:
bath
1
EYE2
Ivcon
drops
1
RES4
5ap
ora
M:INF
1
DER2
34
Ivloc
empl
Hc
1
Medicinal Ethnobotany
2.5 Informants
Table 2 6 List of healers and midwives participating the ethnobotanical project
Name Specialisation Location
Claudia Uc Cahun Parfera hie>rbatera Chikindzonot
Abundio Chan Kauil H-men, hiei'batero partero Chikindzonot
Gregono Cen Uc H men, hier•batero Chikindzonot
Florencio Hoi Chan Hierbatero Chikindzonot
Juanita Pech Balam Parfera Chikindzonot
Maria Pastora Kauil Pech Hieratera Chikindzonot
Juventina Kauil Diaz Parlera Chikindzonot
Claudia Naidelfia Noh Pech Parfera Chikindzonot
Fehpa Moo Kahun Fartera Chikindzonot
Jose Carlos Chan Kauil Curandera Chikindzonot
Vicente de Paul Moo Pat Hierbatero, sobadoro EkpedzWilfndo Poot Moo Herbatero Ekpedz
Regulo Moo Dzib Hierbatero EkpedzRosa Maria Dzib Kauil Hierbatera EkpedzGumersindo Cocom Chan Hierbatero EkpedzAntolina Poot Kauil Parfera EkpedzCeliana Moo Pat Parfera EkpedzJustino Dzib Kauil Hierbatero EkpedzNarcisa Poot Poot Hierbatera EkpedzVicente de Paul Dzib Dzul Curandero, hierbatero EkpedzJuana Paula Moo Dzib Parfera EkpedzSinla Pat Cocom Hierbatera EkpedzJuan Santos Dzib Moo Hierbatero EkpedzNarcisa Poot Poot Hierbatera EkpedzJuana Paula Moo Dzib Parfera EkpedzValentina Pat Hue Parfera Ekpedz
Eligio Pat Carnal, H-men EkpedzAndrea Moo Pat Parfera EkpedzVicente Zim Poot Huesero, curandero EkpedzVicente de Paul Dzib Dzul Curandero, hierbatero EkpedzVacunda Dzib Dzul Parfera, curandera Ekpedz
Felipe Pat Chan Hierbatero EkpedzJustino Dzib Kauil Hierbatero EkpedzPedro Acantara Pat Cocom Hierbatero EkpedzJuan Santos Dzib Moo Hierbatero EkpedzPorfina Cocom Pech Parfera EkpedzJose Jsabel May Poot Curandero Xcocmil
Rajelia Poot Poot Hierbatera Xcocmil
Anselmo Chulm Chi Curandero Thiosuco
Emihana Parfera, Hierbatera Chichimila
Age of the informants 27 - 96 years
60
Medicinal Ethnobotany
2.6 Gardens of medicinal plants
The development of a medicinal plant garden in Chikindzonot and Ekpedz was
planned as a mark of gratitude towards the healers and midwives for their co¬
operation in the project. It was also meant as a sign of support of the use of
medicinal plants. These intentions were successful in some cases in others not.
For instance, the building of two medicinal gardens, one close to the clinic run by
the SSA (Secretana de Salud y Asistencia) in Chikindzonot and the other one in
the garden of the clinic of the traditional healers in Ekpedz, was not continuously
cared for due to organizational and political problems. On the other hand, the
translator, influenced by our work together, created a small medicinal plant garden
close to her home and she was (and still is) invited to the healer organization of the
INI (Instituto Nacional Indigenista). The species of medicinal garden in the CICY
(Centra de Investigacion Cientifica de Yucatan, Mérida) were labeled based on the
information of the healers and midwives of the study region.
Species of the botanical garden in Chikindzonot and Ekpedz:
Aloe vera; Bidens squarrosa; Capraria biflora; Chenopodium ambrosioides;
Cnidoscolus aconitifolius spp. aconitifolius; Crossopetalum gaumeri; Cymbopogon
citratus; Dorstenia contrajerva; Guazuma ulmifolia; Hamelia patens; Malmea
depressa; Morinda yucatanensis; Ocimum micranthum; Piscidia piscipula; Ruellia
nudiflora; Tecoma stans.
Species of the botanical garden in Mérida:
Aloe vera; Anredera vesicaria; Bursera simaruba; Cecropia obtusifolia; Citrus
aurantiifolia; Cymbopogon citratus; Hamelia patens; Malmea depressa; Mentha aff.
piperita; Pilocarpus racemosus; Psidium guajava; Punica granatum; Selaginella
longispicata; Tamarindus indica; Tithonia diversifolia.
61
Medicinal Ethnobotany
2.7 Selection of plant species for their biological evaluations
Several plant species were screened in various bioassays. The goal of this
evaluation was to better understand the use of the medicinal plants and their
pharmacological effects. The selection of the species was based on:
their importance as a remedy among the Yucatec Maya of the study region (the
numbers of the documented use reports)
the healers consensus concerning the plant use and preparation
the endemic occurrence of the species on the Yucatan Peninsula or the original
occurrence of the species in America
the lack of phytochemical and/or pharmacological investigations of the species
the analysis of ethnobotanical literature regarding the species
The results of the biological and pharmacological evaluations are shown in chapter
5 and 6.
62
Publication I
Medical ethnobotany of the Yucatec Maya:
Healers' consensus as a quantitative criterion
Anita Ankli1, Otto Sticher1 and Michael Heinrich2
1) Department of Pharmacy, Swiss Federal Institute of Technology (ETH) Zurich,
Winterthurerstr. 190, CH-8057 Zürich, Switzerland
2) Institute of Pharmaceutical Biology, Schänzlestr. 1, Albert-Ludwigs-University,
D-79104 Freiburg, Germany
Published in
Economic Botany 53 (1999) 144-160
Publication i
Abstract
There is an urgent need to obtain information on the relative importance of a taxon
used medicinally as compared to others within a culture. This was achieved
through a documentation of the current indigenous medical uses of 320 species in
three Yucatec Maya communities during 18 months of fieldwork. The 1,549
indivdual reports documented were divided into nine groups, which classify
indigenous uses. The frequency of usage of the individual plants reported was
employed in the analysis of the ethnobotanical importance of the respective taxa.
Species cited more frequently in a group of indigenous uses are regarded to be of
greater ethnobotanical importance than those cited only by a few informants. In
order to obtain information on possible biological, pharmacological and
toxicological effects of some particularly important species, the scientific literature
on these taxa was evaluated systematically.The study is the basis for
phytochemical and pharmacological evaluations of the traditional uses.
Key Words: Yucatec Maya traditional medicine, indigenous medicinal plants,
ethnobotany, evaluation of indigenous uses, quantitative method, Yucatan
(Mexico).
64
Publication I
Etnobotanica medica de los Mayas Yucatecos: consenso de curanderos
como criterio cuantitativo
Se considéra esencial la documentacion de la importancia relativa que un taxon de
uso medicinal tiene, en comparaciön con otros taxones dentro de una misma
cultura. Con este propösito se realize un estudio etnobotânico de 18 meses,
investigando el uso de 320 especies en très comunidades Mayas del Estado de
Yucatan (Mexico). Se documentaron 1,549 usos indigenas, que se clasificaron en
9 grupos. Se utilize el numéro de usos indigenas para determinar la importancia
relativa de cada especie; asi, las especies médicinales que fueron citadas con
mayor frecuencia se consideran las de mayor importancia, mientras que las
especies citadas con menor frecuencia son las de menor importancia. Para
evaluar los usos indigenas se obtuvo informacion sobre efectos biolögicos,
farmacologicos y toxicologicos de las especies, através de una revision
sistemâtica de la literatura cientifica. Este estudio es la base para la selecciön de
plantas que se evaluaran en estudios fitoqui'micos y farmacologicos.
65
Publication I
Introduction
In recent years we and others called attention to the lack of information on the
relative importance of a medicinal plant (or other useful plant) within a culture and
the need for comparing the uses of plants interculturally (Heinrich, Rimpler and
Antonio B. 1992, cf. Etkin 1994, Moerman 1996). A constructive method to obtain
such information is the quantification of indigenous uses (Phillips 1996) which is
appropriate when the relative importance of each use is similar, as with
pharmaceutical preparations such as medicinal plants used for different types of
illnesses. Accordingly, this paper is the third in a series on Mexican indigenous
medicinal plants (Frei, Sticher and Heinrich 1998 on the Isthmus Sierra Zapotecs,
Oaxaca; Weimann and Heinrich 1997 on the Nahua of the Sierra de Zongolica,
Veracruz). An additional goal of these studies has been the selection of plants for
phytochemical and biological/pharmacological studies (Bork et al. 1997, Kato et al.
1996).
Therefore all three of our ethnobotanical studies use similar methodologies (see
also Methods): (1) Specialists in medicinal plants (for example, healers, midwives,
herbalists) were interviewed during 14-18 months of fieldwork and the use-reports
of each informant recorded. (2) The use of the plants is grouped into 9-10
categories.
The principal groups are similar in all three studies: gastrointestinal disorders,
illnesses of the skin (mostly infections and subsequent inflammatory reactions),
respiratory disorders, gynecological (and andrological) conditions. Since there exist
ethnobotanical differences between the three ethnic groups, 5 - 6 additional groups
were formed, which are only used in one or two of the studies, for example, plants
used for bites and stings of poisonous animals (only Maya), opthalmological
illnesses (Maya and Zapotec), and culture bound syndromes (Nahua and
Zapotecs).
This comparative method facilitates the selection of medicinal plants for phyto¬
chemical and biological/pharmacological studies, and is useful in determining the
ethnobotanical importance of a particular plant in contrast with others in the same
use category (Heinrich et al., 1998). There have been several other approaches to
66
Publication I
establish quantitative criteria for the relative ethnobotanical importance of plants
(Berlin and Berlin 1996, Friedman et al. 1986, Johns, Kokwar and Kimanani 1990,
Phillips 1996). The method of Berlin and Berlin is of special relevance to ours. We
both used a similar approach, however they interviewed the general population and
thus recorded and evaluated an enormous set of positive responses (30,000,
Berlin and Berlin 1996: 81-82). Their method requires a considerable investment in
research funds and personnel. The method presented here is tailored to allow for
the assessment of the relative cultural, medical and, consequently, also the socio-
economical importance of plants employed by medical specialists in an ethnic
group and is feasible with a lower input of resources.
A detailed study of Yucatec Mayan medicinal plant use seemed to be of particular
relevance. Traditional forms of treating illness among the Yucatec Maya of Mexico
who still use locally available plant resources are of considerable importance. Their
medical system and knowledge is without doubt a vital part of their culture
(Redfield and Villa R. 1990 [orig. 1934], Roys 1931, Standley 1930, Steggerda and
Korsch 1943). Although many aspects of Mayan ethnobotany have been
addressed in detail, such as ethnoecology (Herrera 1994, Humphries 1993, Terän
and Rasmussen 1994) and plant nomenclature (Barrera M., Barrera V. and Lopez
F. 1976, Sosa V. et al. 1985), only a few reports exist on the medicinal plants
currently used. For example, Arnason et al. (1980) studied a Mayan community in
Belize, Comerford (1996) a Mayan community in lowland Guatemala, and Alcorn
(1984) the Huastec Maya. In addition, some theses have been written on the topic
and booklets are distributed locally or regionally (Cardeha V. 1985, Pulido S. and
Serralta P. 1993; cf. Mendita and Arno 198, Morton 1981, Argueta V. 1994).
Our study thus has a twofold purpose: (1) to systematically record the use of
medicinal plants in communities of the Yucatec Maya in Mexico, and (2) to form the
basis for comparative studies on Mexican Indian medicinal plant use.
67
Publication I
Background
Yucatan and the Maya
The peninsula of Yucatan forms the easternmost part of Mexico which is divided
politically into three states; Yucatan, Quintana Roo and Campeche. The northern
parts of Guatemala and Belize are also part of the peninsula and have a high
percentage of Lowland Mayan speakers (Fig. 1). The peninsula is an enormous
plateau, made of limestone, with an average altitude of less than 100 m a s I. No
surface rivers exist in the northern part of the peninsula. The most important water
sources are cenotes (natural sink holes formed by the collapse of the limestone
surface over the ground). The annual rainfall is highest in the southeast (1,300 -
1,400 mm) and diminishes towards the north and northwest (400 mm). The natural
vegetation in the southeast is tropical rainforest, while in the extreme northeast it is
low tropical deciduous forest. Due to the low latitude, the climate is warm and in
the southeast, humid.
For more than a millennium the civilizations of the Ancient Maya flourished on the
peninsula and adjoining regions (Koehler 1990). Currently 600,000 persons, or 36
% of the total population of the peninsula, are mono- or bilingual speakers of Maya
(Pfeiler 1995). Even though the influence of outside forces has been enormous, the
Yucatec Maya still retain a large number of ancient traditions and have opposed
the cultural dominance of the Spaniards and the surrounding national Mexican
culture. The so-called "caste war" of the last century which involved the Maya, is
just one example of the resulting conflicts (Orosa D. 1991).
Yucatec Maya language belongs to the mayance (or mayoide) subfamily of
Macropenutian. Maya vowels and consonants are generally pronounced as in
Spanish. A glottal stop ['] is known and glottalized consonants are frequent. In this
article, Maya words are transcribed after Barrera M., Barrera V. and Lopez F.
(1976).
68
Publication I
The Communities
This study was conducted in the communities of Chikindzonot (20° 20' degrees
latitude north, 88° 29' degrees west, 30 - 40 m a s I) and the neighboring Ekpedz
and Xcocmil south of the city of Valladolid in the southeastern part of the state of
Yucatan. These communities were selected because of a high rate of speakers of
Yucatec Maya, the well known cultural conservatism in this region and the high
number of healers known (Orosa D. 1991, Tuz, Valladolid, pers. comm.)
Figure 1. Region of fieldwork
Average annual temperature in the area is 25.7 °C and it has an average annual
precipitation of 1,220 mm. According to Duch (1988) the climatic subtype is Aw
1"(x')(i')g, i.e., a hot subhumid climate with rain from May to October (980 mm) and
little thermal oscillation throughout the year. The vegetation is characterized as a
median semideciduous forest with an average height of 10 - 20 m. 50 to 75 % of
the species remain deciduous during the dry season. Typical tree species include
Acacia pennatula (Schlecht. & Cham.) Benth., Bursera simaruba Sarg.,
Caesalpinia gaumeri Greenman, Cochlospermum vitifolium Willd. ex Spreng.,
Enterlobium cyclocarpum Griseb, Guazuma ulmifolia Lam., Gymnopodium
floribundum Rolfe, Mimosa bahamensis Benth. and Vitex gaumeri Greenman
(Salvador F. and Espejel C.1994).
69
Publication I
The communities of Chikindzonot and Ekpedz have 1,500 and 800 inhabitants,
respectively (INEGI 1990). The whole municipio of Chikindzonot has 2,750
inhabitants. 56% of the persons older than 15 yrs. are literate and one third of
those older than 5 yrs. are monolingual speakers of Maya and the remainder
bilinguals. The economy is based on subsistence agriculture (mostly maize, beans
and squash) and on the production of honey, fruit (watermelon and citrus fruits)
and cattle breeding. Hunting is still practiced regularly, especially by younger men.
Handicraft articles (hammocks and huipiles - female dress) are sold in the market
of Valladolid. No detailed anthropological monograph on the Maya of this area is
available, but the community of Chan Kom, which was first studied by and Villa R.
1990 [orig. 1934], is only 27 km to the north.
Health and healing
According to the unpublished data of the local health authorities and our surveys it
is apparent that gastrointestinal disorders (frequently diarrhea and - in younger
children - as a result thereof, dehydration) and respiratory illnesses are major
health problems. Infected wounds and other inflammatory skin diseases are also
common. Bites from poisonous snakes (e.g., tsab - cascabel - Crotalus durissus L.)
are feared, yet only a few cases have been recorded in recent years. Chronic and
infectious eye diseases are frequent. Diabetes is now considered an important
health problem by the local health authorities, and informants often claim to suffer
from this illness. Most of the medicinal plants sold in the markets of Valladolid and
Mérida promise to alleviate diabetes or to function as diuretics.
The best known group of healers are h-men (Maya for knowledgable healer or
prayer maker), who are not only healers but also specialists in religious rites and
who perform ceremonies addressing the rain-god to ask for protection of the milpa
or the community. He or she is the owner of a sastun, a stone used for divining.
Midwives and herbateros (specialist in medicinal plants) form another group of
healers. Those of the latter group are generally proficient in treating broken bones
and thus work as hueseros. Massages are given by another group of healers - the
sobadores - and by midwives. All these groups of healers make extensive use of
70
Publication I
medicinal plants. Some groups largely use these plants as part of empirical
medications, while others, in particular, the h-men, also use the plants for ritual
purposes.
An outpatient clinic run by the SSA (Secretaria de Salud y Asistencia) and staffed
with a pasante (a medical student in her or his last year of training) and a mestizo
nurse provides biomedical health care, but for most conditions people still prefer
the Mayan healers. In 1993/1994 the pasante (a woman) was called only once to
help during a delivery.
Methods
Ethnomedical and ethnobotanical data were collected mostly in the two
communities of Chikindzonot and Ekpedz from February 1994 until May 1995 and
September/October 1996, however the data from the second stay are not included
here. Specialists in medicinal plants and/or healers of the different regions were
interviewed. This paper is based on structured and unstructured interviews with 40
healers. Twelve healers aged between 29 and 71, who represent all the groups of
specialists mentioned above were interviewed frequently and contributed a large
share of the information presented here. Together with informants we collected
plant materials that were cited as medicinal. During meetings of groups of
indigenous healers, we conducted unstructured interviews on the medicinal plants
and methods of treatment. We thus obtained information on the use(s),
preparation(s), plant parts used, application(s) and properties of the plants as well
as descriptions of illnesses and treatments, which we compiled into ethnobotanical
data sheets.
Voucher specimens were collected and are deposited at the Herbarium of the
"Centra de Investigacion Cientifica de Yucatan" (CICY) in Mérida, the National
Herbarium of Mexico (MEXU), the Instituto Nacional Indigenista (INI) in Valladolid
(both Yucatan), the ETH Zurich (ZT) and the Institut für Pharmazeutische Biologie
in Freiburg, Germany (collection numbers A. Ankli, AANK1 - 540). Plants were
identified by comparison with authentic specimens and in some cases with the
assistance of specialists at CICY and MEXU.
71
Publication I
Reports of ethnobotanical uses were documented for each plant. The healers were
asked to demonstrate the plants which they currently use or which they had used.
In order to analyze the data, the plants were arranged into nine classes of
indigenous use and for each class the data were quantified by adding up the
individual reports on the uses of each plant. Species were then ranked according
to the number of reports of use. Plants in these groups which were cited as
medicinal by four (in case of the gastrointestinal illnesses by five) or more
informants are presented here, A literature search in BIOSIS PREVIEW (and in
selected cases, also in NAPRALERT and Chemical Abstracts) was performed for
the plants most frequently cited in order to obtain data on phytochemistry and on
biological and pharmacological effects of the respective taxa.
Results and discussion
Illness according to the Yucatec Maya may be classified as being humorally "hot"
or "cold" (Redfield and Villa R. 1990 [orig. 1934]). imbalance of the body, for
example, caused by consuming something "cold'' when a person is in a "hot" state
may lead to illness. Diarrhea, for example, is considered to be a "cold" illness,
while dysentery (bloody diarrhea) is classified as "hot". This classificatory system is
also central during and after childbirth. After giving birth, a woman is considered to
be in a "cold" state and therefore is not allowed to eat certain food such as pork,
beef and foods which contain much grease. The "hot/cold" classification of the
Yucatec Maya was recorded by Redfield and Villa R. 1990 [orig. 1934] and was
reported in the sixteenth century (Lopez A. and Viesca T. 1984, Villa R. 1981).
Nonetheless, the humoral system does not encompass the entire medical system
of the people we worked with. Many uses of plants are based on personal
experience or oral transmission within the family.
Illness may be caused by "wind" (i.e., by bad air which enters a weak person's
body), witchcraft or nocturnal birds and bats. Younger informants now refer to
microbes as another causative agent of disease. When a person is treated, ritual
and empirical plant use are closely connected. If the illness requires it, the healer
72
Publication
includes a ritual cleansing ceremony (santigua or limpia). The healer asks God for
help and for the permission to cure.
In this paper we concentrate on the medicinal use of plants among the Yucatec
Maya. We document a total of 1,549 individual responses to queries concerning
320 plant species. These responses were grouped into nine categories of
indigenous use (Fig. 2). We constructed all major and most of the lesser categories
to coincide with indigenous classifications (cf. Berlin and Berlin 1996). A few of the
smaller categories are combinations in order to accommodate, for example, the
various forms of chronic and acute "pain" and illnesses associated with a rise of
body temperature. One residual group ("other uses") is also included. Yucatec
Maya healers are well acquainted with the causes and detectable signs
(symptoms) of illness. It is no surprise then that the groups relate to the organs
affected during a certain illness (gastrointestinal tract, skin, respiratory system).
Figure 2. Quantitative ethnobotanical analysis of the nine groups which classify the
indigenous use reports (total number of species: 320; n = 1549 use reports; AT =
counteract bites and strings of venomous animals; DER = dermatological
conditions; EYE = illnesses of the eyes; FEM = women's medicines; Gl =
gastrointestinal disorders; PFE = illnesses associated with pain and/or fever; OTH
= other uses; RES = respiratory illnesses; UR = urological problems).
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Publication I
The largest number of species and individual use-reports (Fig. 2) were obtained for
gastrointestinal disorders. This group of disorders was second only to
dermatological illnesses in the studies of the Isthmus Sierra Zapotecs and Nahua
of the Sierra de Zongolica (Frei, Sticher and Heinrich 1998., Weimann and Heinrich
1997) in terms of the number of use-reports and among the Zapotecs in terms of
the number of species used. Among the Nahua it was the group with the largest
number of species recorded. Dermatological problems are the second largest
group among the Yucatec Maya. It is noteworthy that these two groups of uses are
prominent in all studies mentioned. Also, respiratory illnesses yield a large number
of use reports and 87 taxa - the third largest number. Of particular importance
according to the Yucatec Maya are plants used in the treatment of bites from
venomous animals, especially snakes (4.8% of all use-reports). In the following,
the principal species used by the Yucatec Maya for each of the 9 groups are
discussed. The ethnobotanical data are summarized in Tables 1 - 7.
Gastrointestinal disorders In this group 147 species with 476 use reports were
documented. Diarrhea is a frequent condition, especially in children. Other
illnesses, included in this group, are gastrointestinal cramps, vomiting and mal de
ojo (evil eye, no Yucatec Maya term used). The latter is an illness caused by a
person, who looks at a child with a so called "strong glance". A drunken person, a
women during menstruation and people born on Tuesday or Friday are particularly
likely to cause this illness. Its signs are various gastrointestinal symptoms,
particularly "green" diarrhea, gastrointestinal cramps and vomiting. To cure the ill
person, the one who has caused the illness has to embrace the patient or to show
her/his care in another way. Additionally herbal preparations are used. Another
important illness is tip'te' (cirro). It is an "organ" reported to sit below the umbilicus.
If a person has eaten inadequate food or after having carried something heavy the
tip'te' palpitates and is dislocated (cf. Berlin and Jara 1993 on a similar concept in
Highland Chiapas). Treatment consists mostly in circular massages around the
navel and in drinking a decoction of tip'te' ak {Microgramma nitida).
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Publication I
The three members of the genus Mentha - M. piperita, M. citrata (leaves/aerial
parts) and M. arvensis (roots and leaves), which are widely used to treat vomiting
and to a lesser degree diarrhea (Table 1), are widely known and effective
antispasmodics of European origin (Haensel et al. 1993). Abrus precatorius is used
only externally for diarrhea and the evil eye in the form of baths and therefore is not
discussed further. Manilkara zapota (mostly bark, also roots and fruits) is employed
for diarrhea and is best known for yielding chicle - a latex rich in polyisoprenes.
The bark is also known to be rich in tannins (Hegnauer 1973:296), but no data on
their structure have been published (Hegnauer 1990: 498). While a huge number
of publications on the production and use of the latex are available, no
pharmacological information relating to the indigenous uses is available. The
relevant reports on the genus Aristolochia were recently summarized (Weimann
and Heinrich 1997). No data are available to substantiate the Mayan uses for
diarrhea. Callicarpa acuminata (shoots) is a Yucatec Maya remedy for mal de ojo
associated with green diarrhea and dysentery. The genus Callicarpa is one of the
few in the Verbenaceae (s.l.) in which no iridoids could be found (Falk 1992).
Instead diterpenoids as well as triterpenoids and flavonoids were reported from the
genus (Glasby 1991). Whether these exert any relevant pharmacological effects is
unknown. Lippia alba, L. stochadifolia (leaves) and related species are used in
many different cultures for gastrointestinal disorders, the Yucatec Maya value the
species to induce vomiting. Many species of this genus are rich in essential oil
{Lippia alba > 1 %; Hegnauer 1973:668) and thus may act as carminatives.
Roots of Dorstenia contrajerva are used for a variety of gastrointestinal conditions,
chiefly for stomach ache, "air in the stomach" and gastrointestinal cramps. Similar
uses of D. drako are reported from the Istmo Sierra Zapotecs (Frei, Sticher and
Heinrich, 1998). Furanocoumarins are known from D. contrajerva (Terreaux et al.
1994), but no data are available on its pharmacological effects. Pimenta dioica is
rich in essential oil (with approx. 35 % eugenol and 40 - 45 % eugenol
methylether), has a wide use for unspecified gastrointestinal problems (List and
Hoerhammer 1977) and may well be an adequate treatment of "air in the stomach".
A central nervous depressant effect of the aqueous (and to a lesser degree of the
75
Publication I
ethanolic) leaf extract applied i.v. was recently shown as part of a hippocratic
screening (Suârez et al. 1997). Psidium guajava (leaves and sometimes also root)
is used for diarrhea and dysentery. These uses are also described by Aguilar et al.
(1994), Weimann and Heinrich (1997) and Giron et al. (1991). The leaves are
mentioned in Farmacopea Mexicana as astringents. Pharmacological tests on the
leaves showed activity against E. coli and spasmolytic effects (Berlin and Berlin,
1994, Weimann and Heinrich 1997) which reportedly are due to a calcium-
antagonist effect of quercetin glycosides (Morales et al., 1994). All plant parts are
rich in phenols, especially hydrolyzable tannins and proanthocyanidins (Okuda et
al. 1987). Teloxys ambrosioides (syn.: Chenopodium ambrosioides) is another
widely used Mesoamerican species employed as anti-emetic, antiparasitic and
antidiarrhetic. Ascaridol is a well-known monoterpene with antiparasitic effects, but
also with undesirable side effects (Hegnauer 1964:421).
Artemisia ludoviciana ssp. mexicana (leaves) has a long tradition of use in
Mesoamerica. The Yucatec Maya value this plant for treating vomiting. It is rich in
sesquiterpene lactones, but no data on anti-emetic effects are available (Bork et al.
1996, Heinrich 1996). Ruta chalepensis (leaves) is known from many cultures and
widely used as antispasmodic. It is prominent for its content of a large number of
different alkaloids and furanocoumarins, is a common abortifacient and has severe
side effects (Haensel et al. 1993, 1994, Heinrich 1989).
76
TABLE
1SPECIESUSEDFORGASTROINTESTINALDISORDERS
(lis
ted
inorderofdecLrvngfrequency
ofuse)
Number
Vouche-'
Species
Family
PlantPart
MayaName
MainUses
of
AANK
Uses
Mentha
aff
piperita
L
indet
AbrusprecatonusL
Manilkarazapota
(L)vRoyen
AristolochiamaximaJacq
Call
icar
paacuminata
Kunth
Lippia
stoechadifolia
(L)Kunth
Teloxysambrosioides
(L)WA
Weber
DorsteniacontrajervaL
Mentha
aff
citrataEhrh
Pimentadioica(L
)Merr
PsidiumguajavaL
Artemisia
ludoviciana
Nutt
ssp
mexicana
(Wil
ld)Keck
Cissampelos
pareiraL
Citrusaurant
i um
L
Lipp
iaalba
(Mil
l)N
EBrex
Britton&
Wilson
MalvaviscusarboreusCav
var
arboreus
RutachalepensisL
BidenssquarrosaLess
Cissus
trifol
iata
(L)Lour
Citrussinensis
(L)Osbeck
Labiatae
Iv,tw,ap
ba,wo
Legummosae
se,
Iv,
fl
Sapotaceae
ba,
rt,fr
Anstolochiaceae
rt,wp
Menta,Balsamoxiw
Canelaycuyo
Oxo
Ya
,Zapote
Chiclezapote
Wahkohak
,Guaco
cast
illo
Verbenaceae
sh
Puk
in
Verbenaceae
kTedechina
Chenopodiaceae
Iv,tw,
rtEpazote,Apazote
Moraceae
rhKambalhaw
Labiatae
VTo
ronj
il
Myrtaceae
Iv,se
PimientadeTabasco
Myrtaceae
rt,
fr,
IvPichi
,Guayava
Compositae
kSi
isim
Menispermaceae
Iv
Rutaceae
W
Peteltun,Orejade
raton
Pak
al,Naranjaagna
Verbenaceae
IvTedelimon
Malvaceae
rt,iv
Bisil,
Holol
Rutaceae
Compositae
Iv,ap
V
Ruda
Ya'xkan-ak'
Vitaceae
Rutaceae
k Iv,se,
pe
fr,
-fr
C^uz
ojo
China,Naranjadulce
vomiting,
parasites
inthestomach
diar
rhea
,dysentery,
a,r
inthestomach
malde
ojo,greendiarrhea
diarrhea,dysentery
diarrhea,dysentery,
air
>nthestomach,
pasma*
malde
ojo,greendiarrhei
dysentery
vomiting
para
site
s,pasma,
vomiting
air
inthestomach
vomitingpasma
air
inthestomach
diarrhea
vomiting,diarrhea
malde
ojo,greendiarrhea
yellowdiarrnca,
air
inthestomach
of
youngerchildren
vomiting
vomiting
dysentery
malde
ojo,mal
viento,diarrhea
malde
ojo,
green
diar
rhea
,vomiting
malde
ojo,
greendiarrhea
air
inthestomach
ofyoungerchildren
18
031
12
256
12
001
12
234
11
350
11
169
11
010
10
025
10
330
10
030
10
023
10
235
9012
9008
9236
9011
9140
9055
8121
8043
8247
-J
-si
co
TriumfettasemitrilobaJacq
ZingiberofficinaleRoscoea
Mentha
aff
arvensisL
OcimummicranthumWilld
PunicagranatumL
mdet
Hylocereus
undatusjxL
)Britton&Rose
Microgramma
nitida
(JSm
)A
Reed
Sm
Piscidiapi
scipula(L
)Sarg
Bauhiniaherrerae
(Britton
&Rose)
Standley&Steyerm
Citrusaurantiifolia(Chnstm
)Swingle
Crossopetalumgaumeri(Loes)L
unde
lI
HeliotropiumangiospermumMurray
PimpmellaanisumL
ZanthoxylumcanbaeumLam
Tihaceae
rtMul-och
dysentery
8230
Zingiberaceae
rhGengibre
indi
gest
ion,
air
inthestomach
8370
Labiatae
rt,ap
Hierbabuena
vomiting,parasites
7029
Labiatae
Iv,ju
-lv
Kakalîun
dysentery
7033
Punicaceae
fr,
IvGranada
yellow
diarrhea
7272
-
Iv,ap
Pasmarxiw
stomachache,cramps,pasma
6422
Cactaceae
kPitahaya
dysentery
6427
Polypodiaceae
wp
Tip'
te'-ak
cirro
tip
te
,stomachache
6183
Leguminosae
sh
Ha
abin
malde
ojo,dysentery
6123
Leguminosae
kKibix,
Ts'ulubtok
malde
ojo,
whitedysentery
5124
Rutaceae
kLimon
airinthestomach
5257
Celastraceae
rtViperolnegro
diarrhea,dysentery
5038
Boraginaceae
IvNema
ax
dysentery,malde
ojo
5053
Apiaceae
sc
Anisengranos
air
inthestomach
5255
Rutaceae
rt,
ivSinanche
mal
vien
to,diarrhea
5368
*
pasma(r)
Oftenmentionedasacause
forgastrointestinal
illnessesand
isus
uall
yassociatedwith
drinksheorshe
gets
ill,
ap=
aerial
part
,ba^bark,
fl=flowers,
fr=
fruits,
ju-fr=juiceof
fruits,
ju-l
=peel
of
frui
t,pet=
peta
ls,
re=
resin,
rh=rhizome,
rt=
root
,se=seeds,
sh=
shoot,
tw=
twig,
stomchacheWhen
apersonma
hotstatetakescold
j=juiceofle
aves
,la=
latex(milky),
Iv=
leav
es,
pe-f
rwo=wood,
wp=whole
plan
t
Publication I
Dermatological Conditions In this group 302 use reports referre to 150 species.
Small cuts, infections and other skin problems are frequent and generally treated at
once with plants (150 species with 302 use reports) that are readily available.
Other frequent health problems are pimples (saa sak' winkli), warts and scabies.
The rootstock of Anredera vesicaria is used as a wound dressing and for infections
(Table 2). Practically no chemical information is available on the genus. Only a
retrochalcone was isolated form another species of this genus (Calzado et al.
1990). Calea urticifolia, another remedy for pimples, contains sesquiterpene
lactones and is an effective inhibitor of the transcription factor NF-kB, which
controls genes responsible for the inflammatory responses of the body (Borges de
C. et al. 1981, Bork et al. 1997), Crossopetalum gaumeri is known to have
antibacterial and cytotoxic effects (see Plants Used to Counteract Venomous
Animal Bites and Stings). Diospyros anisandra is employed in the treatment of
pimples and scabies. Of the approximately 240 species in this genus, only D. kaki
and D. mollis were investigated in greater detail by various groups. Triterpenes
(lupeol, betulin, betulinic acid) and naphtoquinones are widely distributed in the
genus. D. anisandra has yet to be studied (Hegnauer 1989:403-407). The leaves of
Ocimum micranthum and Salvia micrantha are both used for infections and
inflammation of the skin. Neither species has been studied in detail, but O.
micranthum especially merits further study (Heinrich 1992). Species of both genera
are rich in essential oil; in Salvia spp. neo-clerodane diterpenoids were frequently
reported as bioactive compounds (Rodriguez-Hahn et al. 1995). The use of
Kalanchoë intégra (leaves) as a topical anti-inflammatory seems to be based on
the succulent nature of the leaves. Whether the plant exerts any specific anti¬
inflammatory effect is unknown. Leaves of Psidium sartorianum are used for the
same conditions. While no specific data for this taxon are available, species of this
genus are rich in hydrolyzable tannins and proanthocyanidins in all plant parts
(Okuda et al. 1987), and thus may have some anti-infective effects. No
pharmacological data are available to substantiate the claims in the case of
Samolus ebracteatus.
79
coo
TABLE
2SPECIESUSEDFORDERMATOLOGICALCONDITIONS
Species
Family
Plant
Part
MayaName
Pnmulaceae
kTs
unya
iSBasellaceae
rtKaa'xicheel
Crassulaceae
kBelladona
Compositae
kKaxikin
Celastraceae
rt.l
vVi
pero
lnegro
Ebenaceae
kKakalche',Kanan
Labiatae
Iv,ap
Kakaltun
Myrtaceae
IvPichiche1
Labiatae
Iv,ap
Ya'xkax
Rubiaceae
ap
Haway
Orchidaceae
kCh
itkuuk
Euphorbiaceae
la.l
vEk'balam,Xikmburro
Rubiaceae
kElelkabil,
Kanan
Euphorbiaceae
kXul-im-il
Euphorbiaceae
kXilkin,Kambaikiche'
Apocynaceae
laUtsupek'
Simaroubaceae
kBelsinikche'
Leguminosae
kAzufre-xiw
Rubiaceae
fr,
IvPiha
ak'
Phytolaccaceae
frT'elkox
Myrtaceae
kPi
chi'
,Guayaba
Punicaceae
kGranada
Leguminosae
kSaal,Saalche'
Cucurbitaceae
kChakmotz-ak',Hoyke
MainUses
Number
Voucher
of
AANK
Uses
9032
8196
8016
6153
6038
6134
6033
6211
6025
5276
5188
5205
5199
5357
5088
5190
4136
4125
4113
4388
4235
4272
4084
4036
Samolus
ebracteatusKunth
Anredera
vesicariaGaertner
f
Kalanchoë
inté
graKuntze
Calea
urti
cifoliaMi
llsp
varyucatanensis
Wussow,Urb&
Sullivan
Crossopetalum
gaumeri(Loes
)Lundell
DiospyrosanisandraBlake
OcimummicranthumWilld
Psidiumsartonanum
(Ber
gius
)Nied
Salvia
micranthaVahl
Borreria
verticillata
(L)G
Mey
CatasetumintegernmumHook
CrotonperaeruginosusCroizat
HameliapatensJacq
PhyllanthusacuminatusVahl
PhyllanthusmicrandrusMeuII
Arg
Tabernaemontana
amyg
dali
foli
aJacq
AlvaradoaamorphoidesLiebm
Daleaca
rthagenensis
var
barbata
(Oerst
'
Barneby
MonndayucatanensisGreenman
Phytolacca
icosandra(L
)Sims
PsidiumguajavaL
PunicagranatumL
Senna
villosa
(Mil
l)Irwin&Barneby
Sicydiumtamnifolium(Kunth)Cogn
wounds,
inflammation
inflammation
tumor,
inflammation
litt
le,redpimples
x-onob
inflammation
oftheneck
itch
ingpimples
usan
pimples,whitespots
siklikmuch'
measles
k'aa'ahko
lebill
scabies
pimples
x-onob
smal
l,whitepimple
bigpimples
chukum
pimples
inflammation,redpimples,
wounds
inflammation
oftheni
pple
s
wound,
infe
ctio
n,inflammation
of
theneck
warts
itch
ingpimples
pimples
wounds
measles
pimples
saasak
wmkli
measleskaaah
kolebil
hard
,li
ttlep'mples
AbbreviationseeTable
1.
Publication I
Illnesses Associated with Pain and/or Fever This category (see Table 3) is
rather diverse and includes plants (112 species 204 with use reports) that are used
for chronic or acute pain and fever. These ailments are commonly treated with
external appplications of species known to be rich in essential oils (e.g. Ruta
chalepensis, leaves, aerial parts, locally known as an external remedy for
headache, see above). Illnesses associated with fevers are generally treated with
baths. Pain is generally treated with plasters and other external applications.
Satureja brownei [syn.: Micromeria brownei {Sw.) Benth.; leaves] is used in the
form of baths for headaches and has a diverse range of uses among many other
peoples in Central America, the Caribbean and Mexico (Heinrich 1992). Essential
oil and methylated flavones are found in taxa from this genus (Tomas B., Husein
and Gil 1988). The ethanolic leaf extract showed inhibitory activity against
Staphylococcus aureus and Streptococcus pyogenes in a screening of 68
Guatemalan plants (Caceres et al. 1991), but no relevant pharmacological data are
available on the species used and therefore it is not possible to evaluate the
Mayan use. Zanthoxylum caribaeum (roots) is widely known among the Yucatec
Maya as a cure to rheumatism, headache, and "mal viento". Many species of this
large genus with approximately 200 species have been studied phytochemically
and/or pharmacologcially in detail. Characteristic are benzyl isochinolin alkaloids,
lignans, coumarins and unusual amides (Hegnauer 1990: 449-455). Since the plant
is employed externally in ritual cleaning ceremonies, an evaluation of the
indigenous claims goes beyond the scope of this study.
Ocimum basilicum is a well known species rich in essential oil (0.5 - 1.5 %)
introduced into numerous indigenous medical systems of the Americas (Heinrich
1992). Analgesic or antibacterial effects would be of relevance for the Yucatec
Maya use in case of toothache, and there is some information on antibacterial
effects of the essential oil (Heinrich 1989, List and Hoerhammer 1977). Leaves of
Bursera simaruba are exclusively employed externally for fever (baths). The genus
is rich in essential oils, Triterpenes, lignans and procyanidies are also found
(Khalid 1983), but are probably of no relevance to the use discussed here. Its
81
Publication I
external use for fever is known from other parts of Mexico and seems to be
partially based on the aromatic smell of the leaves, resin and bark (Heinrich 1989).
Respiratory Illnesses The species in this group (87 with 177 use reports) are
locally used for cough, bronchitis and "asthma" (Table 4). The latter illness is
described as a longlasting wheezing, caused by winds called tus ik'. In
phytotherapy in many European countries plants rich in essential oils or in
polysaccharides are regarded as useful therapies for the first two conditions (Ph.
Helv. 8). Only Cymbopogon citratus (leaves) is known to be rich in essential oil
(usually employed as a spasmolytic). The genus Ehretia is rich in rosmarinic acid,
which has antiviral, antihistamine release and anti-inflammatory effects (Kuhnt et
al. 1995, Simpol et al. 1994). Other important Mayan medicinal taxa are Croton
lundellii (leaves), Euphorbia ptercineura (latex), Rosa chinensis (leaves) and
Turnera diffusa (leaves). Some species might have moderate antibiotic effects, but
there are no data to further substantiate the Mayan claims. No chemical data on
these taxa are available that would substantiate the indigenous claims, but some
species {Croton lundellii, Euphorbia ptercineura) might have severe side effects,
because of the presence of phorbole esters.
82
TABLE
3.SPECIESUSED
INTHETREATMENTOFILLNESSESASSOCIATEDWITHPAINORFEVER
NumberVoucher
Species
Family
Plant
Part
MayaName
MainUses
ofAANK
Uses
Satureja
brownei(S
w.)
Briq.
Labiatae
Rutachalepensis
L.
Rutaceae
ZanthoxylumcaribaeumLam.
Rutaceae
Ocimumbasilicum
L.Labiatae
Burserasimaruba
(L.)
Sarg
.Burseraceae
Blxaorellana
L.Bixaceae
BunchosiaswartzianaGriseb.
Malpighiaceae
Cestrumnocturnum
L.
Solanaceae
Ehretia
tini
foli
aL.
Boraginaceae
Hybanthusthiemei
(F.Donn.Sm.)Morton
Violaceae
UreracaracasanaGriseb.
Urticaceae
Iv,ap
Poleo
headache
Iv,ap
Ruda
headache,
trembling
rt,Iv
Sinanche'
ma!
vien
to,feverrheumatism
ap
Albahaca
headache,toothache
Iv,re
Chakah
feverwithacoldbody:
k'ilkab
sh
Kiwi,K'uxub,Achiote
feve
r,trembling
ofbabies
ap,tw
Sipche'
headache,malde
ojo,
fever
kJuandenoche
feverwithacoldbody:
k'ilkab
Iv,ba
Beek,
Roble
fever
ap
Fluxion'xiw
toothache
kLaal,Or
tiga
rheumatism
8044
8055
8368
7071
6042
4233
4142
4050
4021
4266
4325
TABLE
4.SPECIESUSEDFORRESPIRATORYILLNESSES
NumberVoucher
Species
Family
Plant
Part
MayaName
MainUses
of
AANK
Uses
CDco
Croton
lundelliiStandley
Cymbopogon
citratus(Nees)Stapf
Ehretia
tini
foli
aL.
EuphorbiaptercineuraA.Berger
Rosachinensis
L.
Turnera
diffusa
(Willd.exSchultes)
Bauhiniadivahcata
L.
Citrusaurantiifolia(Christm.)
Swingle
Gossypiumhlrsutum
L..
Pseudobombax
ellipticum
(Kunth)Dugand
Piscidiapisc
ipul
a(L.)
Sarg
.Cornutiapyramidata
L.
Murrayapa
nicu
lata
Jacq
.
Euphorbiaceae
Iv
Gramineae
Iv
Boraginaceae
ba,
Iv
Euphorbiaceae
la
Rosaceae
Iv
Turneraceae
k
Leguminosae
Iv,
fl
Rutaceae
Iv,ju
-fr
Malvaceae
Iv,
fr,
fl
Bombacaceae
Iv,ba
Leguminosae
sh
Verbenaceae
Iv
Rutaceae
fl,tw
Kok-ché
Zacatede
limön
Beeb,
Roble
Xmuch
kok
Rosa
Oreganodemonte
Mayvaca,Patadevaca
Limön
pais
Chuytaman,Algodon
Amabola,
Xk'unche'
Ha'abin
Xolte'xnuk
Limonaria
asthma:
tus
ik',
bron
chit
is,cough
asthma,cough,
catarrh
cough,asthma
asthma,cough
cough
cough,
bronchitis
asthma,cough,
bronchitis
cough,asthma,
bronchitis
cough,asthma:
tus
ik'_
asthma,cough,
bronchitis
cough,asthma
feve
r,asthma
asthma,
bronchitis
7040
7061
7021
7034
7130
6193
5007
5257
5086
5275
4123
4048
4099
AbbreviationseeTable
1.
Publication I
Gynecological Uses Table 5 shows the most frequently mentioned species out of
74 species with 129 use reports. Plants used during delivery are the most
prominent group in this category. As described above the categorization into "hot"
and "cold" illness and remedies is culturally important. Infertility of the women is
regarded as a cold illness and consequently "hot" remedies (e.g. Pluchea
symphytifolia) are prescribed. Infertility is considered an important problem by the
healers.
Bark and leaves of Guazuma ulmifolia are used to relieve the pain of childbirth. It is
one of the best known plants in the two communities and has practically no other
uses. Among the Lowland Mixe it is used as a remedy for diarrhea and also for
pain in the uterus and vaginal hemorrhages (Heinrich 1989), among the Isthmus
Sierra Zapotecs for diarrhea and fever (Frei, Sticher and Heinrich, 1998).
Polymeric proanthocyanidins are common and antisecretoric effects on cholera
induced colonic secretion were reported (Hoer, Rimpler and Heinrich 1995). No
data are available to validate the therapeutic claims of the Yucatec Maya. Pluchea
symphytifolia (leaves) is mostly used to "warm up the womb", for "irregular
menstruation" and uterine spasms. According to the Yucatec Maya the preparation
of the remedy is essential for the therapeutic effect. If an abortive effect is desired,
the remedy has to be drunk while it is still hot. For the other therapeutic uses the
remedy is drunk when it has cooled down. While some data on the biological
effects of the plants and on biologically active caffeoylquinic acids are available
(Scholz, Heinrich and Hunkler 1994), no information validates the traditional Mayan
uses. No data are available which support the reported use for relieving pain of
Pisonia aculeata.
Plants Used to Counteract Venomous Animal Bites and Stings In this group 44
species with 76 use reports were documented. Most of the plants are applied
topically on the wound caused by a snake or scorpion. Many persons state that the
treatment of snake bites requires 16 species, but no informant listed that many
taxa. Crossopetalum gaumeri is the most frequently mentioned species in this
group. All informants report that immediately after the bite of a snake one should
84
Publication I
chew a piece of the drug. The powder drug is also applied externally and/or orally
as a decoction. Triterpenes of the oleanan, lupan amd friedelan type are frequently
reported in the Celastraceae and also in the genus. Characteristic for the family are
chinoid pigments - the celastroids - which are derived from friedelanes. These
compounds are reported to have antibacterial and cytotoxic effects (Hegnauer
1989:223). Anredera vesicaria and Urechitis andrieuxii are wound dressings. No
data on antivenomous effects of any one of these species are available.
Urological Problems In Table 7, 44 species with 66 use reports are summarized.
"Kidney trouble" is mentioned most frequently, yet it does not represent a specific
condition. Many of the plants may act as diuretics, the Yucatec Maya term for the
illness - k'aluix - is generally explained as "the patient can't pass the urine". Also
included in this group is "diabetes". The Yucatec Maya consider plants to be
effective in the treatment of the latter illness if the plants act as diuretics. Malmea
depressa - the most popular treatment of "kidney trouble" among the Yucatec
Maya - was recently investigated phytochemically in detail for the first time and
revealed the presence of phenylpropanoids (Jimenez A. et al.1996). No data to
substantiate the indigenous claims are available.
Eye Remedies In this group 39 use reports referred to 27 species. Inflammations
and disturbing, long lasting spots on the surface of the eye (buy) are frequently
treated with herbal remedies. Often drops prepared from the leaf-sap of various
plants are used and applied topically. No plant stands out as particularly more
important than the others: Ocimum basilicum L, Chamaechsta glandulosa Greene
and Desmanthus spp. are mentioned three times each, Euphorbia hirta L. and E.
heterophylla Desf. two times. In the case of 0. basilicum the mucilaginous seeds
are used for the treatment. No attempts were made to validate the indigenous
uses.
85
Co
CD
TABLE
5.WOMEN'SMEDICINES
NumberVoucher
Species
Fami
lyPlant
Part
MayaName
MainUses
of
AANK
Uses
Guazuma
ulmifoliaLam.
Sterculiaceae
ba,
ivNohoch
pixoy
Pisoniaaculeata
L.Nyctaginaceae
sh
Beeb,Unadegato
Plucheasymp
hyti
foli
a(M
ill.)
Gillis
Compositae
kChalche'
Pimenta
dioica
(L.)
Merr.
Myrtaceae
Iv,
frNukuch
pool
,Pimientade
Tabasco
AristolochiamaximaJacq.
Aristolochiaceae
itWahk'oh
ak'
indet.
Ulmaceae
ba
Kamba
pixoy
Plmpinella
anisum
L.Apiaceae
se
Anisengranos
Zuelanlaguidonia
(Sw.
)Britton&
Flacourtiaceae
itBotox,Tamay
Millsp.
childbirth,abortion*
childbirth,abortion
desire
ofhaving
achild,
pasmo0,
abortion
pain
,co
lic,
problems
ofmenstruation,pasmo,
bath
ofvagina
problems
ofmenstruation,pasmo
childbirth,abortion
childbirth
desire
ofhavinga
child,pasmo
12
250
7154
7009
6023
5350
5248
4255
4375
*
The
inductionorprevention
ofabortiondependson
thedosage
(high,
low)
and
ofthepreparation
(hot,co
ld);
"painduring
menstruation,darkblood,
infe
rtil
ity.
AbbreviationseeTable
1.
TABLE
6.SPECIESUSEDFORINJURIESCAUSEDBYVENOMOUSANIMALS
Species
Family
PlantPart
MayaName
MainUses
Number
of
Uses
Voucher
AANK
Crossopetalumgaumeri(Loes.)Lundell
Anredera
vesicaria_Gaertner
f.
UrechitesandrieuxiiMuell.Arg.
Celastraceae
Basellaceae
Apocynaceae
rt,iv
rt rt,Iv
Vipe
rolnegro
Kaa'xiche'el
Vipe
rolbejuco
snakebite
snakebite,
snakebite,
wound
inflammation
12 6 5
038
196
466
AbbreviationseeTable
1.
TABLE
7.SPECIESUSEDFORUROLOGICALPROBLEMS
NumberVoucher
Species
Family
Plant
Part
MayaName
MainUses
of
AANK
Uses
Malmeadepressa
(Bâillon)R.E.Fries
Annonaceae
Chromolaenaodorata
(L.)
R.M.King&
H.Rob.
Compositae
Bauhiniadivaricata
L.Leguminosae
Cecropia
obtusifolia
Bertol.
Moraceae
Parmentieraaculeata(Kunth)Seemann
Bignoniaceae
rtElemuy
diur
etic
,kidneytr
oubl
e,kidney
stones
rtTok'aban
diabetes,kidney
trou
ble,
urine
does
notpass:
k'alwix
Iv,
rtMayvaca
kindney
trouble
Iv,
rtK'
ooch
le',
Guarumbo
diabetes
rt,fr
Kat
diabetes,
urinedoes
notpass:
k'alwix
8161
6339
4007
4041
4135
AbbreviationseeTable
1.
c»
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Other Uses The uses summarized in this group (53 species with 80 use reports)
are very diverse. But only two plants stand out as being of some importance. In
most cases it is not possible or meaningful to evaluate the therapeutic claims.
Piper amalago (6 use reports) is popularly used to fight dandruff and split hair tips.
No attempt was made to ascertain the therapeutic value. The rootstock of
Anredera vesicaria is used for broken bones as a hard gypsum like plaster
bandage.
Conclusion
This study clearly demonstrates that medicinal plants still are an important natural
resource for the Yucatec Maya of Chikindzonot, Ekpedz and Xcocmil. However,
biomedical pharmaceuticals are becoming increasingly popular. Such
pharmaceuticals are of less importance in the area we studied as compared to
other Yucatec Mayan communities. No detailed study on these forms of treatment
in the Yucatec Maya area exists (van der Geest, Reynolds and Harden 1996).
Most of the species recorded in this study have been reported as useful medicinals
in other regions of Central America, Mexico and the Caribbean, but contrary to
these studies we have presented data that allow the evaluation of the relative
importance of a particular species in the medical system of the Yucatec Maya. The
method also is useful for other culturally important plant products (e.g. dyes). It is
based on interviews conducted by a single researcher, with the help of a field
assistant or translator. In our previous studies with the Zapotecs and Nahua, we
documented 3,600 and 800 individual use-reports, respectively. (Frei, Sticher and
Heinrich 1998, Weimann and Heinrich 1997). These use reports yield relevant
information on the intracultural importance of a specific plant as compared to other
taxa. In a subsequent analysis the three ethnobotanical studies conducted with a
similar methodology will be compared.
The quantitative data have to be seen in comparison to other types of
ethnobotanical information. Garden plants are generally more important in the
medical system of the Yucatec Maya than plants collected outside the community.
Many informants also proudly show (newly) introduced taxa (esp. Mentha spp.,
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also Citrus spp., Lippia alba). Consequently, in order to foment the indigenous use
of medicinal plants a medicinal plant garden is being built up in cooperation with
two of the three communities. Other types of information relating to the cultural
importance of a plant can be drawn from the classification of plants in the
indigenous system (the humoral system or systems based on organoleptic
properties of plants; Heinrich n.d). Such a discussion goes beyond the scope of
this paper.
Based on estimates by Bye (1993:707) 15 % or 5000 species of the Mexican flora
are used medicinally. The method presented here contributes to the selection of
the most important ones, which then can be studied pharmacologically,
toxicologically and phytochemically in order to evaluate the indigenous claims
(Robineau and Soejarto 1996). These studies will also help to revalue the
indigenous uses of medicinal plants. Plants which seem to be of particular
relevance for such studies are, for example, Anredera vesicaria, Dorstenia
contrajerva, Diospyros anisandra, Guazuma ulmifolia and Zanthoxylum caribaeum.
Studies on some of these plants are consequently under way.
An earlier version of this paper focusing on plants used in gastrointestinal medicine
was presented at the symposium "Plants for Food and Medicine", July 1 - 6, 1996,
Imperial College, London at the joint meeting of the Society for Economic Botany
and the International Society for Ethnopharmacology.
Acknowledgments
This research would not have been possible without the collaboration of the
healers, midwives and other inhabitants of the communities we worked in, who are
the traditional keepers of this knowledge. The botanical identification at CICY and
MEXU was performed in collaboration with the numerous specialists of this
institution. Particularly we would like to thank Dra. I. Olmsted, J. Granados, P.
Sirna and J.C. Tejön of CICY as well as 0. Tellez, R. Lira and Dr. M. Sousa of
MEXU. This research owes a lot to the continued support of Prof. H. Rimpler
(Freiburg), and to the help of Dra. B. Pfeiler (UADY, Mérida), Dr. Tuz (INI,
Valladolid) and Dr. Baltisberger (Zürich). We are very grateful to Dr. John Plant
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(Freiburg) for critically revising the manuscript. Financial support by SDC (Swiss
Agency for Development and Cooperation, Berne, Switzerland) and the SANW
(Swiss Academy of Natural Sciences) is gratefully acknowledged.
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Yucatec Maya medicinal plants versus nonmedicinal
plants: Indigenous characterization and selection
Anita Ankli1, Otto Sticher1 and Michael Heinrich1,2
1) Department of Pharmacy, Swiss Federal Institute of Technology (ETH) Zurich,
Winterthurerstr. 190, CH-8057 Zürich, Switzerland
2) On leave from: Institut für Pharmazeutische Biologie, Albert-Ludwigs-Universität,
Schänzlestr. 1, D-79104 Freiburg, Germany,
Fax.:+49-761-203-2803
Published in
Human Ecology 27 (1999) 557-580
Publication II
Abstract
Medicinal plants are an important part of the environment as it is perceived by
Mexican indigenous groups. The aim of this study, which was conducted over a
period of 18 months in three Yucatec Mayan communities, is to better understand
the selection criteria for medicinal plants. An important group of selection criteria
are the flavor and aroma of plants. The absence of smell or taste indicates that the
taxon has no potential medical value. Medicinal plants are more often considered
to be sweet or aromatic (to smell good) or astringent, while a similar percentage of
medicinal and nonmedicinal plants are considered bitter, spicy, acidic, or bad
smelling. The relationship between the ethnobotanical data obtained for the
individual plants and the secondary plant products (natural products) prominent in
each species is specifically addressed in this paper. It shows that an understanding
of the indigenous concepts used to distinguish medicinal from nonmedicinal
species has considerable heuristic value.
KEY WORDS: Indigenous knowledge; medicinal plants; nonmedicinal plants;
traditional medicine; ethnobotany; plant selection criteria; taste; smell; hot-cold
classification; Yucatec Maya; Yucatan (Mexico).
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Desde el punto de vista de los grupos indigenas Mexicanos, las plantas
médicinales son una parte relevante del entorno natural donde estos se
desarrollan. El objetivo de este estudio se centra en el entendimiento del o los
criterios que el grupo indigena Maya Yucateco utiiiza para seleccionar una planta
como medicinal. El trabajo se realize durante 18 meses en très comunidades
Mayas Yucatecas. Cuando se compararon las plantas médicinales contra las
plantas no-medicinales se encontre que las caracteristicas de sabor y olor son un
criterio para la seleccion de una planta como medicinal. Asi, la ausencia de sabor
o olor indica un potencial valor medicinal. Se encontre que las plantas médicinales
son consideradas frecuentemente como dulces, aromâticas (con olor agradable) 6
astringentes, siendo para el caso de las plantas no-medicinales las caracteristicas
de amargo, picante, acido y con olor agradable los aspectos que constituyen el
criterio de selecciön.
Se analiza de manera especi'fica la relaciön entre los datos etnobotânicos
obtenidos para cada planta y los productos naturales secundarios de las mismas;
ésto, con el objetivo de mostrar que el entendimiento de los conceptos indigenas
usados para distinguir las especies de plantas médicinales de las no-medicinales
es de un considerable valor heristico.
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Introduction
Impressive collections of documented plants used in indigenous medical systems
are available (for the Americas see, for example, Aguilar et al., 1994; Argueta V.,
1994; Heinrich, 1996; Moerman, 1998; Morton, 1981), but the ethnobotanical study
of medicinal plants has largely remained descriptive, and the process and rationale
for the use of plants as medicine has not been explored in detail (Brett and
Heinrich, 1998). Researchers in the field have generally been more interested
towards the practical application of ethnobotanical information in Western
biomedicine or its uses in primary health care1 (Balick and Cox, 1996; Robineau
and Soejarto, 1996). Among the most noteworthy exceptions are the work of Etkin
(e.g. 1994), Moerman (1996), and Johns (1990).
On the other hand, theoretically informed ethnobotanical studies have made major
contributions to related fields such as ethnoecology (Alcorn, 1984; Balée, 1994;
Ellen and Fukui 1996), cognitive anthropology, ethnoscience (Berlin 1992), and the
study of humoral classification of such disparate phenomena like food, types of
illnesses and plants (Foster, 1994). Foster's study is particularly relevant in the
context of this paper. Some authors have systematically explored the hot-cold
concept and its role in indigenous medicine and diet (Foster, 1994), but little
information is available on botanically identified species and their classification in
this system (cf. Messer, 1991). Also, we have recently criticized the hot-cold
system as too narrow to explain plant use (Brett and Heinrich, 1998). The selection
of plants as medicine based on their taste and smell seems to be important, but
hitherto little explored in many cultures (Brett and Heinrich, 1998 and references
therein; Crellin and Philpott, 1997). That taste and smell are particularly important
criteria for characterizing medicinal plants has been shown in ethnobotanical
1 Such approaches are exemplified by many of the articles published in journals such as the Journal of Ethnopharmacology,
Fitoterapia, Economic Botany and Pharmaceutical Biology (formerly International Journal of Pharmacology).
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studies with the Tzeltal and Mixe (Brett, 1998; Heinrich, 1998). Those two studies
did not, however, look at the differences in the people's taste and smell perceptions
of medicinal versus nonmedicinal plants, the specific focus of this paper. The
cultural reasons for selecting a plant as a medicinal one, the cultural processes
which allow the selection of new medicinal plants, the forms of transmission of this
knowledge, and the management of culturally important plants in the environment
(e.g. by growing them in the house yard or sparing a plant during the cleaning of an
area; Heinrich, 1997) are pertinent topics for ethnobotanical research on medicinal
plants.
The Maya of the Yucatan peninsula (Mexico) are a particularly relevant example,
since plants are an integral and important part of their indigenous culture and since
they have strongly resisted outside influences. Traditional treatments for illness
among the Maya of the Yucatan peninsula, who still use locally available plant, are
of considerable importance. Detailed studies of their medical system and
knowledge (Redfield and Villa R., 1990) and of many aspects of their ethnobotany,
including ethnoecology (Herrera O, 1994; Terân and Rasmussen, 1994) and plant
nomenclature (Barrera et al., 1976; Sosa et al., 1985) are available. Only a few
reports address currently used medicinal plants (cf. references cited in Ankli et el.,
1999). In this paper we analyze the Yucatec Mayan2 criteria for distinguishing
between medicinal and nonmedicinal plants. The wealth of ethnobotanical
information of the Yucatec Mayan healers as well as the intra- and intercultural
variations in medicinal plant knowledge has been described and analyzed before
(Ankli et al., 1999; Heinrich étal., 1998). In this paper, the hot-cold classification
and the classification system based on taste and smell among Yucatec Maya are
2Unless stated otherwise the term "Yucatec May a" is used throughout this paper to refer to the inhabitants of three
communities south of Valladolid (see Background), since no common denominator exists for these three communities. If we
make reference to the Yucatec Maya in general, we use the term Maya of the Yucatan peninsula.
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examined to discover how the Maya distinguish between medicinal and
nonmedicinal plants.
The Study Area and the people
Yucatan and the Maya
The land of the lowland Maya stretches over most of the peninsula of Yucatan,
which contains the easternmost part of Mexico and the northern parts of
Guatemala and Belize. The peninsula is an enormous limestone plateau; its
highest altitude barely reaches 400 ms above sea level (Hernandez, 1985). No
surface rivers run through the northern part of the peninsula; the most important
water sources are cenotes (natural sinkholes formed by the collapse of the
limestone surface over the ground). The annual rainfall is greatest in the southeast
(1,300 - 1,400 mm) and diminishes towards the north and northwest to 400 mm.
The southeast portion is tropical rainforest, and in the extreme northeast low
tropical deciduous forest. The latitude and the adjoining warm sea make the
climate warm and humid.
Yucatec Maya language belongs to the Mayance (or Mayoide) subfamily of
Macropenutian. Maya vowels and consonants are generally pronounced as in
Spanish. A glottal stop ['] is used, and glottalized consonants are frequent.
Currently 600,000 persons, or 36 % of the total population of the peninsula, are
mono- or bilingual speakers of Maya (Pfeiler, 1995). In this article, Maya words are
transcribed after Barrera et al. (1991).
This study was conducted, in the communities of Chikindzonot and the neigh¬
boring Ekpedz and Xcocmil, south of the city of Valladolid in the southeastern part
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of the state of Yucatan. Average annual temperature in the area is 25.7 °C and the
average annual precipitation 1,220 mm. It is a hot, subhumid climate that has rain
from May to October (980 mm) and little temperature variation throughout the year
(Duch, 1988). The vegetation is characterized as a median semideciduous forest
with an average height of 10 - 20 m. Some 50 % to 75 % of the species remain
deciduous during the dry season (Salvador and Espejel, 1994). The communities
of Chikindzonot and Ekpedz have 1,500 and 800 inhabitants, respectively (INEGI,
1990) and the whole municipio of Chikindzonot has 2,750 inhabitants (Fig.1). Fifty-
six percent of the people over 15 years of age are literate and a third of those over
five years are monolingual speakers of Maya, the rest being bilingual. The
economy is based on subsistence agriculture (maize, beans, and squash) and on
the raising of honey, citrus fruits, watermelons and cattle.
Fig. 1. Place of field study.
The most important group of healers are h-men, who are not only healers but also
specialists in religious rites and who perform ceremonies asking the rain god for
protection for the milpa (com field) or the community. He or she is the owner of a
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sastun, a stone used for divining. Midwives {ah-k'ax tuch ahal: person who cuts
the navel string) and herbalists {ts'a ts'ak xiw: person who gives medicinal plants)
form another group who are generally proficient in treating broken bones.
Massages are given by another group of healers - the masseur {yet)- and
midwives3. While all these groups of healers make extensive use of medicinal
plants, some use them largely as part of phytotherapeutic preparations while
others, in particular, the h-men, also use the plants for ritual purposes.
No detailed anthropological monograph on the Maya of this area is available, but
the community of Chan Kom, which was first studied in the 1930s by Redfield and
Villa R. (1990) is only 27 km to the north.
Methods
Ethnomedical and ethnobotanical data were collected from February 1994 until
May 1995 and in September/October 1996, mainly in the two villages of
Chikindzonot and Ekpedz. The information was gathered from structured and
unstructured interviews with 40 traditional healers and midwives. Twelve healers
aged 40 - 71 years were interviewed frequently and contributed much of the
information presented here. Ten of these were between 40 and 50 years of age,
and all actively practice their healing art. All twelve consider themselves herbalists,
and most of them also work as h-men (2), masseurs (5), midwives (4) or bone
setters (3). Generally, the midwives also work as masseurs. Six healers are men
and six women.
Together with informants, we collected voucher specimens and additional material
of medicinal taxa. During meetings of groups of indigenous healers, we conducted
unstructured interviews on the medicinal plants and methods of treatment. We
3The Maya terms for the various groups of empirical healers generally are descriptive.
104
Publication II
thus obtained information on the use(s), preparation(s), plant parts used,
application(s) and properties of the plants as well as descriptions of illnesses and
treatments, which were compiled into ethnobotanical data sheets.
Reports of ethnobotanical uses were documented for each plant. The healers were
asked to indicate which taxa they were currently using or which ones they had
used. For each species, information on the plant part(s) used, their preparation and
application, and the Maya uses, plus data on the indigenous classification of the
species, were recorded. To analyze the data, the use reports for a species were
arranged into nine groups based on symptoms or application techniques
mentioned by the healers (Ankli et ah, 1999; Heinrich et al., 1998). For each group,
the data were quantified, adding up the individual reports on the uses of each
plant. They were then ranked according to the number of reports of use (see Ankli
et ai, 1999).
To distinguish between medicinal and nonmedicinal plants we looked at plants
considered by a single informant (generally a healer) to be medicinal and
compared them to those plants the same person considered not useful as
medicine. After having completed the documentation of medicinal plants of the
communities, individual healers were asked to select a maximum of 10
ethnobotanical taxa that in their opinion had no medical value. The healers were
next asked about the taste and smell, the humoral, and other properties of the
taxa. The informants generally were unfamilier with the taste and smell of taxa they
did not use. Therefore, they were tasted by the informant, the translator, and the
key investigator. The healer then gave his opinion on the taste and smell properties
of each taxon. Voucher specimens were collected and are deposited at the
National Herbarium of Mexico (MEXU), the Herbarium of the Centra de
Investigacion Cientifica de Yucatan (CICY) in Mérida, the Instituto Nacional
Indigenista (INI) in Valladolid, Yucatan, the ETH Zurich (ZT), and the Institut für
105
Publication II
Pharmazeutische Biologie in Freiburg, Germany (collection numbers A. Ankli 1 -
540). They were identified by comparison with authentic specimens and in some
cases with the assistance of specialists at CICY and MEXU.
The selection of medicinal plants
The informants gave various reasons why a plant was considered to be a
medicine. For many informants, the traditional knowledge passed on from one
generation to the next was a key to deciding why they used a certain plant. Most
of them were taught by an experienced healer {h-men) or by elderly relatives. In
the initial interviews on medicinal plants, a traditional healer reported that good-
smelling plants were useful against stomach ache, bitter-tasting ones against skin
problems, and sweet plants to strengthening the body and the blood. The
reasoning of other healers was not always that a plant was a medicine because it
has a certain taste or smell property but often that the plant was used for an illness
and that it was bitter, astringent or aromatic, etc. The informants did not recall
whether the information on the plants' characteristics was also transmitted to them,
but it seems likely that they learned both what plants to used and, implicitly the
reasons for their use. Other information was obtained through dreaming about
useful plants and subsequently testing the effect of these plants in treatment.
Some plants were selected because they showed similarities to a certain illness or
the diseased body or organ. The flower of Matelea yucatanensis (Standi.)
Woodson (Asclepiadaceae) resembles the navel. Therefore, a single flower is used
against pain of the navel of the babies and for trembling of babies. Because of its
spines, which "hold back the fetus", the aerial part of the cactus pitaya4
4 To facilitate the reading of the text, common English terms for species (following Morton, 1981) have been added in the
main body of the paper if possible (but not in the tables) along with their botanical names. Indigenous names were recorded
and can be found, for example, on the voucher specimens.
106
Publication II
[Hylocereus undatus (L.) Britton & Rose, Cactaceae] are used for preventing
abortion. The fruit of Godmania aesculifolia (Kunth) Standi. (Bignoniaceae)
resembles the umbilical cord and is therefore used to speed up labor and to expel
the placenta.
Based on the work by Redford and Villa Rojas (1990), we had anticipated that
humoral concepts would be important in the selection of medicinal plants. Our
informants never mentioned hot or cold properties of a plant as a reason for its
medicinal use. When asked about these properties they first made reference to the
disease and its humoral characteristics, which this remedy would be used to treat,
and then volunteered data on a plant's property. Additionally, in explaining the
causes of an illness, thermal concepts5 were frequently mentioned by the healers6.
Plants are not chosen at random for treating a certain illness. Sweet plants are
clearly preferred for treating respiratory illnesses; 65 % of all use reports in the
group "respiratory illnesses" concerning taste properties are about plants
commonly considered to be sweet (Table I). Another important characteristic of the
plants in this group is that they typically have a strong odor (65% of all taxa with
ascribed smell properties). Plants for the treatment of bites of venomous animals,
especially snakes, are frequently mentioned (n=14 answers) and generally are
classified as bitter (n=10; 71%). Smell is not an important criterion in this group (n
= 1). Women's medicine are considered to have no smell (40%) or no taste (57%)
or to be aromatic (55 %). For skin conditions, bitter (64%) and astringent (33 %),
5There is a methodological problem in distinguishing between the thermal and humoral properties of a species or an illness.
Frequently the informant refer to a "hot" or "cold'1 item without it becoming clear immediately which of the two types they were
referring to. The data in the following discussion are therefore based on systematic inquiries into the meaning of the terms of
the infoimant.
6Water of the cenotes is considered to be cold. Cool water should not be drunk or used for washing or bathing. Caution must
also be observed with cold winds coming from the North (Maya; cascac ik). On the other hand, the hot sun is feared as a
cause of illness.
107
Publication II
and aromatic (47%) taxa are preferred. The various gastrointestinal illnesses were
divided into three subgroups: dysentery, diarrhea and vomiting. For vomiting,
aromatic plants (61%) or bitter (58%) ones are preferred. Diarrhea generally is
treated with astringent (50%), aromatic (65%), or bitter (33%) plants, whereas the
most popular type of plants for dysentery are those that have only minimal smell, if
any, (67%) or a bitter taste (35%). Pain and fever are treated mostly with aromatic
(52%) or bad-smelling plants (43%).
Responses were sought on the classification of the plants in the various groups of
indigenous uses according to the humoral system (Table II). Disorders specific to
women are treated with remedies considered to be hot (88 % of all responses).
Pain and fever, on the other hand, call for cold ones (74 %). A particularly
interesting group is gastrointestinal disorders. Roughly half of all reports in this
group classify the taxa as hot or cold, respectively. But if one looks, for example, at
dysentery, it is clear that it requires remedies that are cold (96 %). On the other
hand diarrhea and vomiting call for hot remedies. Dysentery is considered to be a
hot illness because of the blood in the feces. Diarrhea and vomiting are caused by
cold winds, cloudy skies, or by rain during the rainy season. Consequently, these
two illnesses are cold and require hot remedies. Inflammatory skin diseases and
bites of venomous animals produce a localized reddening of the skin and
temperature elevation and thus require remedies that are cold.
The main groups of skin conditions distinguished are inflammation and pimples.
Inflammation is considered a hot illness; pimples a cold one. For respiratory
illnesses, the classification of the plants is not clear-cut: 53% of the plants are said
to be hot and 47% cold.
108
Table
I.Classification
ofMedicinal
PlantsbySensory
CharacteristicsandGroup
ofuse:
Taste(%)
Smell(%)
Group
ofuse"
Bitter
Astr
ing¬
Sweet
Tasteless
Spicy
Acid
Aromatic
Odorless
Strong-
Bad-
ent
"tot
smelling
smelling
"tot
Gastrointestinal
42
29
715
43
73
56
26
11
788
disorders
Dysentery
35
17
926
49
23
17
67
-6
15
Diarrhea
33
50
13
4-
-24
65
24
66
17
Vomiting
58
19
-
15
8-
26
61
16
16
756
Dermatological
64
33
3-
--
33
47
720
27
15
conditions
Women's
medicine
29
--
57
14
-
14
55
40
-
520
Painandfever
25
-
25
25
25
-
452
-5
43
21
Respiratoryillnesses
917
65
--
923
24
665
617
Bitesbyvenomous
71
21
7-
--
14
-_
-
(1)
1
animals
a
Figu
reswithshadedbackground:>30%,>3
responses.
bUrological
problems^
tot=
1°I
otherusesn^
=2;
conditionsoftheeyesn
^0^
:
Publication II
Table II. Hot-Cold Classification of Medicinal Plants in the Groupsof Uses3
Hot Cold
Group of usea (%) (%) /7tot
Gastrointestinal disorders 65 45 101
Dysentery 4 96 27
Diarrhea 67 33 18
Vomiting 77 23 56
Dermatological conditions 29 71 35
Infection 4 96 24
Pimples 82 18 11
Women's medicine 88 12 25
Pain and fever 26 74 23
Respiratory illnesses 53 47 19
Bites by venomous animals - 100 13
a
Figures with shaded background: >70°'o of the total number of
individual use reports; two responses: Urological problems; conditions
of the eyes; other uses.
Differences between medicinal and nonmedicinal plants
Individual healers were asked about differences between medicinal and
nonmedicinal plants. Practically all medicinal plants are named folk taxa. For 28 %
of the nonmedicinal plants, the informants did not know the names of plants that
they did not personally use.
Most of the nonmedicinal taxa are trees and herbs, and they usually are rather
inconspicuous plants, without showy flowers and good-tasting fruit, for example.
When selecting a plant to be discussed as a nonmedicinal, the informants in most
cases described and tasted the leaves. Since the Maya clearly distinguish between
bok (smell) and kii (taste) (Barrera et al., 1991), the answers were recorded in two
groups (Fig. 2 and 3). There is a significant difference between the two groups with
respect to taste and smell properties (%2 = 56, 99.9% CI of %2 is 20 with 5 degrees
of freedom).
Smell was generally described as being good, bad, or absent. In explaining the
smell properties, the informants additionally compared the odor with common ones,
such as the smell of a person, of a lemon, or honey, or the strong, aromatic smell
of adrue {Cyperus articulatus L, Cyperaceae). According to the Maya, a strong
110
^
CE
60
50
40
30
20
10
n169 Responses on
Medicinal Taxa
m 100 Responses on
Non-Medicinal Taxa
Aromatic Odorless Strong smell Bad smell
Quality of Smell
Fig. 2. Smell of medicinal and nonmedicinal plants of the Yucatec Maya.
50
45
40
35
30
25
20
15
10
5
o *•-•
166 Responses on
Medicinal Taxai
m 100 Responses on I
Non-Medicinal Taxa1
I—WM
itter Astringent Sweet Tasteless
Quality of Taste
Spicy Acid
Fig. 3. Taste of medicinal and nonmedicinal plants of the Yucatec Maya.
m
Publication II
smell can be pleasant or unpleasant, but it is distinguished from other groups of
smell by being unusually strong. The ways in which the Maya characterize taste
and smell of a plant were used in the analysis (i.e., we accepted the criteria as they
were stated by the Maya [Table III], including somewhat ambiguous categories
such as "strong" for bad smell).
We have documented a total of 335 responses about taste and smell properties
and 222 about the humoral classification for 329 medicinal taxa. For the 69
nonmedicinal ones, 200 responses concerning sensory perception and just two
responses for humorally hot were recorded (Figs. 2 and 3).
Nonmedicinal plants were more often reported to have no smell (Fig. 2) or no taste
(Fig. 3). Medicinal plants, on the other hand, are more often aromatic (good smell),
and there is practically no difference in the frequencies of responses about bad or
strong smell (Fig. 2). A good odor was mentioned in 50 % of the cases as a
characteristic of a medicinal plant and thus is a sign for medicinal use, whereas the
absence of smell indicates that the taxon has no potential medicinal value.
With respect to taste, a larger percentage of medicinal plants were reported to be
astringent or sweet, but there are no differences in the qualities bitter, spicy, and
acid (Fig. 3). It is noteworthy that the informants considered 44 % and 42% of the
nonmedicinal plants and medicinal plants, respectively, to be bitter. This
characteristic is therefore attributed to a rather large segment of the surrounding
flora but seems to be of no direct relevance to the selection of medicinal plants.
It is worth repeating that, for the Maya, it is not the case that medicinal plants are
characterized as unusually bitter. There is no difference in the percentages of
medicinal and of nonmedicinal plants that are considered to be bitter. This
contradicts reports in other ethnobotanical studies that bitterness is a particular
characteristic of many medicinal plants (e.g., Heinrich et al., 1992).
112
Table
III.
Qualities
ofMedicinalandNonmedicinal
Plants
oftheLowlandMaya
Maya
Spanish
English
Example
Taste
k'a
Amargo
Bitter
suts'
Astringente
Astringent
ch'uhuk
Dulce
Sweet
pap
Picante
Spicy
pu
pah
Agrio
Acid
kl'ubok
Buen
olor
Aromatic,goodsme
chee
ol
Olor
tuerie,apestoso
Strong
smell
tu'ubok
Malolor
Bad
smell
:-cold
chokô
Caliente
Hot
sis
Frio
Cold
Crossopetalumgaumeri
(Loe
s.)Standi,
Psidiumguajava
L.
Pachyrhizuserosus
Urb.
var.palma
Piperamalago
L.
Citrusaurantiifolia
Swingle
Psidiumguajava
L.
Chenopodiumambrosioides
L.
Zanthoxylumcaribaeum
Lam.
Dorsteniaco
ntra
jerv
aL.
Citrusaurantiifolia
Swingle
Publication II
The humoral properties of the plants were also elucidated in the interviews. The
informants sometimes did not clearly distinguish between thermal and humoral
characteristics. Questions about the humoral properties of a plant made no sense
to some of the informants, who proceeded to describ the process of preparing the
remedy (i. e., hot or cold extraction). Frequently, the humoral connotations of hot
and cold describes the illness. The medicinal plant to be used must have the
opposite property (i.e., cold for a hot illness). This does not refer to the selection
criteria for a plant, but to its classification. It therefore comes as no surprise that
nonmedicinal plants are generally not classified humorally (Table IV). In only two
cases was a nonmedicinal one considered to be humorally hot: one {Senna sp.)
because of the Mayan name chak sal (red; little pimples), since red is considered
humorally hot, and the other {Sabal sp.), because the healer was told to use this
species as a women's medicine, nearly all of which are hot.
Table IV. Hot-Cold Classification of Medicinal and Nonmedicinal
Plants of the Lowland Mayas
Responses on Responses on
medicinal taxa nonmedicinal
Classification ntot (%) taxa ntot
Hot 104(46.8) 2
Cold 101 (45.5) _
Cool 12(5.4) __
Lukewarm 5 (2.2) -
Perception and chemical constituents of the plants
The documented sensory perceptions of the medicinal and nonmedicinal plants
were further analyzed using published information on the known chemical
constituents of the species. With this analysis, we intend to reach a better
understanding of the relationship between the indigenous perception of a plant's
taste or odor and the chemical constituents. Tables V and VI show all the plants
114
Publication II
with three or more reports on taste or smell properties and the known (groups of)
constituents. (To facilitate the reading of the text, the plant families and the authors
are listed in the tables).
Taste. It comes as no surprise that species considered to be astringent contain
polyphenols (hydrolyzable tannins and/or proanthocyanidins). The astringent and
disinfecting properties of the polyphenols makes plausible their use for infections
like diarrhea and skin disease.
Bitter-tasting plants are quite common, and bitterness may be attributable to
various groups of compounds such as cardenolides {Dorstenia contrajerva,
Urechites andrieuxii). Other bitter-tasting compounds are terpenes, like the neo-
clerodane-type diterpenes of Salvia spp. or quassin, a quassinoid found in many
taxa of the Simaroubaceae {Alvaradoa amorphoides), and the sesquiterpene
lactones found in many Compositae, such as Calea urticifolia. The genus
Callicarpa is one of the few in the Verbenaceae (s.l.) in which no bitter iridoids
have been found. Instead, bitter diterpenoids, triterpenoids, and flavonoids are
known from this genus. Crossopetalum gaumeri has a bitter taste due to
cardenolides (Ankli et ah, submitted). Naseberry {Manilkara zapota) has latex rich
in polyisoprenes, but no information is available on compounds, which may
produce the bitter taste. Alkaloids are responsible for the bitter taste {Casearia
corymbosa). For soap tree {Sapindus saponaria) saponins or flavonoids are the
relevant compounds.
The yam bean tuber {Pachyrhizus erosus) has a sweet taste probably attributable
to glycoproteins or the flavonoid pachyrhizin. The acid amides of Jamaika black
pepper {Piper amalago) are responsible for its typical spicy taste, like that of black
pepper {Piper nigrum). A typical acid-tasting plant is acid lime (in leaves and fruit
of Citrus aurantiifolia), which contains acidic derivatives of limonoids and acidic
phenols in
115
TableV.MedicinalandNonmedicinal
Plants
ClassifiedbyTasteandSmell
Prop
erti
esAccording
toYucatecMayan
Healersandthe
Constituents
oftheseTaxa3,
Plantname
(familyf
TA
EQ(%)
SL
DT
AK
CM
Other
Ref
Astringent
plant
PsidiumguajavaL,MRT
Manilkarazapota
(L.)
vanRoyen,SPT
Crossopetalumgaumeri(L
oes.
)Standi.,CEL
Punicagranatum
L.,PUN
Bitter
plant
Crossopetalumgaumeri(Loes.)Standi.,CEL
Manilkarazapota
(L.)
vanRoyen,SPT
Dorsteniacontra
jerv
aL.
,MOR
AlvaradoaamorphoidesLiebm.,SMR
Galea
urti
cifo
liaMillsp.var.yucatanensis,
CMP
Callicarpa
acuminataRoxb.,VRB
CaseariacorymbosaJacq.,
FLC
Salviamicrantha
Desf
.,LAB
Sapindussaponaria
L.SAP
+H,
P
+ + +Iv
,fr
+ +
+ +
UrechitesandrieuxiiMuell.
Arg.
,APO
Sweet
plant
PachyrhizuserosusUrb.var.palma,LEG
Spicy
plant
Piperamalago
L.,PIP
+rt
Acid
plant
CitrusaurantiifoliaSwingle,
RUT
+fr
JatrophagaumeriGreenman,EUP
+
Manilkarazapota
(L.)
vanRoyen,SPT
+
PsidiumguajavaL,MRT
+H,
P+
+ +
Lv,
fr
Polyisoprenes
Triterpenes,
sugaralcohols
Lv:tr
iter
pene
s,st
eroi
ds,
fr:sugars,
phenolicacids
Triterpenes,sugaralcohols
Polyisoprene,
fr:sugar
+FU
Cardenolides
Anthraquinones,
trit
erpe
nes(quassinoids)
Phloroglucins
Trit
erpe
noid
s,flavonoids
+Saponins
Terpenoids,
flavonoids
Saponins,
flavonoids,
lipids
Cardenolides
Rt:gl
ycop
rote
ins,
flavonoids(p
achy
rhiz
in)
Piperamides,
sesquiterpenes
+Iv
Terpenes
(lim
onoi
ds),
organicacids
Saponins,
cyanogeniccompounds
Polyisoprene
Lv.fr
12
14,
9
89
39
39,
8
79
79,
3
69,
23
39,
5,
20
39,2
35
39
317,
21
39,
11,
12,25
39
36
49,1
29
29
29
214
,9
Table
V.
(continued)
Plantname
Aromatic
smelling
plant
Psidiumguajava
L.,MRT
Chenopodiumambrosioides
L.(T
eloxys
ambrosioides),CHN
Dorsteniaco
ntra
jerv
aL.
,MOR
Artemisialudovicianassp.mexicana
(Wil
ld.)
Keck,CMP
CitrusaurantiifoliaSwingle,
RUT
Lippia
stoechadifoliaKunth,VRB
Lippia
albaN.
E.
Br.ex
Britton&
Wilson,VRB
Mentha
spp.,LAB
Satureja
brownei(Micromeriabrownei(S
w.)
Benth),LAB
OcimummicranthumWùld.,LAB
Strong-smelling
plant
Chenopodiumambrosioides
L.(Teloxys
ambrosioides),CHN
CitrusaurantiifoliaSwingle,
RUT
Piperamalago
L.,PIP
AlvaradoaamorphoidesLiebm.,SMR
Bad-smelling
plant
ZanthoxylumcaribaeumLam.,RUT
Colubrinagr
eggi
var.yucatanensis,RHM
Senna
uniflora(P.
Mill
er)H.
Irwin&
Barneby,
LEG
Bitter
plant
without
smell
AcalyphaunibracteataMuell.Ar
g.,EUP
TA
EO
SL
DT
AK
CM
Other
Ref.
+P
+(-
2)
+(f
r:0.
4)+
(>1)
+(>1)
+H)
+(-
4)
+(-
2)
+(f
r:0.
4)+
+IA
Lv,
fr
Saponins,
trit
erpe
nes,
organic
acid
s,
flavonoids
FU
Cardenolides
Triterpenes,xanthophylls,flavonoids
IvTerpenes
(lim
onoi
ds),
organicacids
Phenolics
(ros
mari
nic
acid
),flavonoids
Phenolics,
iridoidgl
ycos
ides
,flavonoids
Phenolics,flavonoids
Saponins,
trit
erpe
nes,
organic
acid
s,
flavonoids
+Iv
Terpenes,organicacids
Piperamides
Anthraquinones,
trit
erpe
nes(quassinoids)
Lignans,unusualamids
Saponins,flavonoids
Anthraquinons,
prot
eins
,aromatic
compounds,
flavonoids
Phenolics,cyanogeniccompounds
sugars
12
14,9,
10
99
89,23
59,
18
57,
9
59
49
421,
8
424,
9
37
49
39,
7
39
39,
5,
20
59,
4
39
39
Table
V.
(continued)
Plantname
TA
EO
SL
DT
AK
CM
Other
Ref:
Plants
without
specific
tasteans
smell
AcalyphaalopecuroidesJacq.,EUP
AbutilonpermolleSweet,MLV
TriumfettasemitrilobaJacq.,
TIL
Hylocereusundatus
(L.)
Britton&Rose,CAC
SapindussaponariaL,SAP
+
Microgramma
nitida
(J.Sm.)A.Reed,PLG
+
Phenolics,cyanogeniccompounds
sugars
5/5
Mucilage,
trit
erpe
nes,
fatty
oil,
phenolic
acids
Mucilage,cyanogeniccompounds,
flavonoids
Flavonoids,betalain
Saponins,
flavonoids
Triterpenes,
cyanogeniccompounds,
sugars,
resin
4/4
9
4/4
9
3/4
9
3/4
9
3/3
9
aTA:
tannins;EO,
essential
oils
;SL,sesquiterpene
lactones;DT,diterpenes;AK,
alkaloids;
CM,
coumarins;
H:hydrolyzabletannins;
P,proanthocyanidins;
Bl
benzylisoquino
line
alka
loid
s;1A
,indolealka
loid
s;FU,furanocumarins;
Iv,leaves;
fr,fruits;
rt,roots.
bAbbreviations
ofplantfamiliesfollowthecode
ofWeber,W.
A.(1
982)
.cReferences:1Achenbach
et
al.(1984).2
Borges
delCastilloet
al.(1981).3
Chung
et
al.(1997).4
DeliaCasaandSojo
(1967).
5
Glasby
(1991).cGomes
et
al.
1997
).7Guenther(1949).8
Hansel
et
al.(1993).9
Hegnauer
(1962-1996).10Ji
et
al.(1991
).11Lemos
et
al.
(1992).VlLemos
etal
(1994).nMollonbeck
et
al.
1997).
uOkuda
et
al.(1
987)
.15
Patitucciet
al.(1
995)
.16Rodn'guez-Hahn
et
al.(1
995)
.1/Ruiz-Cancino
et
al.
(1993).
!aScholz
et
al.
(1994).nSchratz
(196
6).2
0SteineggerandHansel
(199
2).2
1Tanabe
et
al.(1
992)
.22Terreaux
et
al.(1995).23Tomas
et
al.
(1988).24WahabandSelim
(198
5).
Publication II
the fruit. Jatropha gaumeri contains caustic latex, which consist of di- and
triterpenes. Especially the unripe fruit of the guava {Psidium guajava) contain little
sugar and are rich in fruit acids. No data are available on naseberry (M. zapotä)
which is also said to be sour.
Smell. All species mentioned as aromatic plants contain relatively large amounts
of essential oils, which are volatile, odoriferous mixtures of compounds that are
largely insoluble in water. The Labiatae (e.g., Mentha, Satureja, Ocimum spp.) and
Myrtaceae {Psidium) are typical representatives of the essential oil-containing plant
families. Two species of the genus Lippia (Verbenaceae) were mentioned several
times by Maya as being aromatic. This genus is one of the few in the Verbenaceae
whose species are rich in essential oil. American wormseed {Teloxys
ambrosioides), Citrus spp., and also estafiate {Artemisia ludoviciana ssp.
mexicana) are mentioned several times as having a good smell, and are all rich in
essential oil (Table V). Dorstenia contrajerva is reported to contain coumarins, but
no data on its essential oil content are available.
The strong-smelling medicinal plants are a puzzling group. American wormseed
{Teloxys ambrosioides) and acid lime {Citrus aurantiifolia) have a strong smell
because they contain much essential oil. Piper amalago and Alvaradoa
amorphoides, on the other hand, are not remarkably fragrant, but their strong and
very characteristic taste seems to be culturally interpreted as strong smell. In this
group a judgment as to the sensory perception seems to be made by the
informants when tasting or smelling the plant. Accordingly, no specific group of
natural products is responsible for a "strong odor".
Among bad-smelling plants, no special group of constituents are responsible for
the unpleasant smell. Scorpion tree {Zanthoxylum caribaeum) is said to smell like
pork, a feature that can probably be attributed to the essential oil or the unusual
amides present in this genus. Senna uniflora is particularly prominent for being rich
119
Table
VI.TypicalHumorallyColdandHotPlantsAcoordmg
toYucatecMayan
Healersand
theConstituents
ofTheseTaxa3
Plantname
TA
EO
SL
DT
AK
CM
Other
Ref
Cold
plant
CitrusaurantiifoliaSwingle,
RUT
+f
r
Hylocereusundatus
(L)Britton&Rose,CAC
Triumfettasemitnloba
Jacq
,TIL
MalvaviscusarboreusCav
var
arboreus,MLV
Crossopetalumgaumeri(Loes
)Standi,CEL
f
Manilkarazapota
(L)vanRoyen,SPT
+
Psidiumguajava
L,MRT
+H,P
+
Artemisialudovicianassp
mexicana
(Willd
)+
+
Keck,CMP
Microgramma
nitida
(JSm
)A
Reed,PLG
++
Satureja
brownei(Micromenabrownei(Sw
)+
Benth),LAB
Hot
plant
DorsteniacontrajervaL
,MOR
ChenopodiumambrosioidesL
(Teloxys
+
ambrosioides),CHN
Triumfettasemitnloba
Jacq
,TIL
Plucheasymp
hyti
foli
a(M
ill
)Gills,CMP
+
Psidiumguajava
L,MRT
+H,P
+
Artemisialudovicianassp
mexicana
(Willd
)+
+
Keck,CMP
Aristolochiaspp
,ARS
CitrusaurantiifoliaSwingle,
RUT
-r
fr
Microgramma
nitida
(JSm
)A
Reed,PLG
++
Pimenta
dioica(L
)Merr,MRT
+
UreracaracasanaGriseb
,URT
+
ZingiberofficinaleRoscoe,Z1N
+
+lv
Terpenes
(lim
onoi
ds),
organicacids
Flavonoids,betalames
Mucilage,cyanogeniccompounds,
flavonoids
Mucilage,fl
avon
oids
,phenolic
acids
Trit
erpe
nes,
sugaralcohols
Polyisoprenes
Lv.fr
Triterpenes,xanthophylls,flavono'ds
Triterpenes,
cyanogeniccompounds,
sugars,
resin
Phenolics(rosmannic
acid
),indo'd
glycosides,flavonoids
+FU
Cardenolides
Saponins,
trit
erpe
nes,
organicacids,
flavonoids
Mucilage,cyanogeniccompounds,
flavonoids
Caffeoylquinic
acd,
flavonoids,simple
phenols,
phenolic
acids
Triterpenes,
xanthophylls,f'avonoids
Anstolochicac
id,anstoloiactame,
lign
anods
+lv
Terpenes
(lim
onoi
ds),
organicacids
Triterpenes,
cyanogeniccompounds,
sugars,
resin
Mucilage
Rt
galano'actons
pheno'ic
acid
s,resin
10
9
89
79
59
49
49
414,9
49
39
324,9
12
9,23
79
79
619
,9
514
,9
49
39
39
39,
15
316
39
313
,22
aTA
tannins,EO,
essential
oils
,SL,sesquiterpene
lactones,
idin
s,Bl
,benzylisoquinoline
alka
loid
s,IA,indolealkaloids,
FU
bAbbreviations
ofplantfamiliesfollowthecode
ofWeber,W
DT,di
terp
enes
,AK,
alka
loid
s,CM,
coumarins,H
hydrolyzabletannins,P
proanthocyn-
,furanocumanns,
Iv,leaves,
fr,fr
uits
,rt
,roots
A(1982)
References-seeTableV
Publication 11
in anthraquinones, but an aromatic smell is not a prominent characteristic
according to the perception of the principal investigator. No essential oil has yet
been reported from Colubrina greggi. Since these three species are not necessarily
very rich in essential oil, the "bad" property seems to be an unpleasant feeling
when tasting the plant. Nearly all plants said to have no smell are also said to be
tasteless. The only exception is Acalypha unibracteata, which has no smell but
reportedly tastes bitter. Practically all plants in this group are known to contain
flavonoids, and some contain cyanogenic compounds and/or tannins.
Based on these data, several clear-cut correlations becomes apparent between
indigenous perceptions and groups of natural products (e.g., aromatic and
astringent compounds). In other cases, no such correlation is noticed. Plants
considered to be aromatic generally contain essential oil, while, for example, plants
described as astringent contain hydrolysable tannins and/or proanthocyanidins.
Hot-cold classification. We were unable to identify any specific group(s) of
compounds associated with the alleged hot or cold properties of a plant (Table VI).
Interestingly, five of 17 species were described by some informants as hot and by
others cold. Thus, agreement among the informants is much lower for the hot-cold
classification.
Conclusions
The aim of this study is to better understand the selection criteria of medicinal
plants used by the lowland Maya. As far as we know, no study comparing
medicinal and nonmedicinal plants nor any that deals specifically with
nonmedicinal plants exists. Our study shows that an understanding of the concepts
indigenous people used to distinguish medicinal from nonmedicinal species has
considerable heuristic value. Sampling the secondary plant products by tasting and
smelling them yields culturally defined clues about a species' potential value and
121
Publication II
helps them to distinguish between used and non-used plants. The approach
employed in this paper thus sheds new light on the selection criteria of the Maya of
the Yucatan peninsula. Plants are not selected at random, nor are they selected
purely by abstract criteria, like hot and cold.
The taste and odor of medicinal plants and the labels applied to them (e.g.,
astringent, bitter, aromatic) include, or encode, considerable information about the
groups of illnesses a particular phytomedicine is best used to treat. Examples
among Yucatec Maya and other cultures are plant remedies classified as
astringent or aromatic. It is, on the other hand, noteworthy that in this study, there
was no difference in the percentage of species classified as bitter in the medicinal
and nonmedicinal plant groups. This contradicts the assumption common in our
culture that medicine has to be bitter. Therefore, no particular sensory property
definitively characterizes a medicinal plant. Taste and smell are very important
selection criteria, but they are not the central unifying principle of indigenous Maya
medicinal plant classification. Indeed, such a unifying concept does not exist (cf.
Worsley, 1997).
In the Yucatec Maya understanding of medicinal plants, specific sensory
properties, like smell, taste, color, form, and texture are the first criteria for
selecting a medicinal plant. These characteristics presumably also serve as
mnemonic aids for identifying medicinal plants that are in regular use. If a plant is
to continue being used as a medicine, it must show a "positive health effect", as
that is interpreted culturally. Therefore, not all plants that share a certain property
serve the same purpose.
Because smell and taste can characterize typical groups of natural products
another key interest here is the relationship between the ethnobotanical data
obtained for the individual taxa and the secondary plant products (natural products)
prominent in each species. This is not only essential for the recognition and
122
Publication II
selection of medicines but can also help to explain the pharmacological effects of a
species (Ankli et al., 1999). Examples are astringent plants, which are widely used,
also among Yucatec Maya, against gastrointestinal disorders and dermatological
problems. The pharmacological effects of these plants, which contain polyphenols,
can partly be explained by the natural products characteristic of the species. Such
a clear-cut connection exists for a few classes of compounds (especially astringent
compounds). Bitter-tasting compounds, on the other hand, are distributed among
a large variety of groups of natural products, so no specific pharmacological
conclusions can be drawn from the fact that a given plant tastes bitter.
Nevertheless, such information is an additional criterion for selecting plants for
phytochemical and pharmacological analysis. Examples are the two species of
Acalypha. The informants consider one of these to be bitter {A. unibracteata),
whereas another {A. alopecuroides) is said to have neither taste nor smell. A
detailed phytochemial comparison of the two would thus be of interest. Since the
data in the scientific literature do not allow quantification of the relevant groups of
natural products, no definite conclusions on the physiological effects of the species
used medicinally can be drawn. In future phytochemical studies, it may be of
interest, not to search for the active principle(s) with an assay for a certain
pharmacological (e.g., anti-inflammatory) activity as a lead but to identify what
compounds are responsible for the taste or smell properties reported by the Maya,
Shifting the focus from the valuable to the "non-valuable" plants gives new
information on the hot-cold dichotomy. In Yucatec Mayan culture, this parameter is
not very important. For our informants, it is a mnemonic aid and an explanation of
plant use and selection for certain illnesses, when the therapy is already known.
Generally, what a species is used for is primary, and, once this is known, the
humoral characteristics are assigned. People who have had regular and more
123
Publication II
intensive contact with Mestizo culture also seem to rely more heavily on this
system than informants who have had less contact.
The analysis presented here also raises a large number of methodological and
conceptual questions. Comparative data on the classification by taste and smell
from other cultures, especially from South America, are urgently needed. Thus, it is
essential that future fieldwork in South America and Mesoamerica not focus on the
hot-cold system or purely on botanical documentation of indigenous medicinal
plants. It will be also interesting to look at the development of classificatory
systems over time to detect changes in the systems. Changing the perspective
from the medicinal to the nonmedicinal plants raises serious questions about our
definitions of medicinal plants. Is a medicinal plant one that is widely used in a
culture and that has been shown to have the pharmacological effects desired by
the informants? What about plants used by very few people or ones that have no
known pharmacological effects? Are these species also medicinal? What
distinguishes a medicinal from a nonmedicinal one? A better understanding of the
multiple ways medicinal plants are classified and thus distinguished form
nonmedicinal will shed light on the differences between the two groups.
Acknowledgments
We are very grateful to the healers, midwives and the inhabitants of Chikindzonot,
Ekpedz and Xcocmil, Yucatan, for their collaboration, for their friendship and
hospitality. This manuscript has profited much from electronic and personal
discussions and other input from Prof. Dr. D. Moerman (Dearborn, Mi). The
botanical identification at CICY and MEXU was performed in collaboration with the
numerous specialists of these institutions. Particularly, we would like to thank Dra.
I. Olmsted, J. Granados, P. Simâ, J.C. Trejo, Dr. R. Durân of CICY as well as O.
124
Publication II
Tellez, Dr. R. Lira, Dr. J. Villaserior and Dr. M. Sousa of MEXU. This research
owes a lot to the help of Dr. B. Frei (Zürich), Prof. H. Rimpler (Freiburg), Dra. B.
Pfeiler (UADY, Mérida), Dr. Tuz (INI, Valladolid), Dr. C. Viesca (UNAM) and Dr.
Baltisberger (Zürich). We are very grateful to S. Ritt for the English revision of the
manuscript and to R. Fisullo and Y. Fang for their help in statistical analysis.
Financial support by SDC (Swiss Agency for Development and Cooperation,
Berne, Switzerland) and the SANW (Swiss Academy of Natural Sciences) is
gratefully acknowledged.
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Yucatec Mayan medicinal plants:
Evaluation based on indigenous uses
Ankli Anita3, Heinrich Michael11'5, Bork Peterb, Wolfram Lutzc, Bauerfeind Peter0,
Brun Retod, ScHMiDCéciled, Weiss Claudia0, Bruggisser Regina', GERTSCHJürg1,
Wasescha Michael8, Sticher Ottoa*
a Department of Applied BioSciences, Institute of Pharmaceutical Sciences, Swiss
Federal Institute of Technology (ETH) Zurich, Winterthurerstr. 190, CH-8057
Zürich, Switzerland
b Institute of Pharmaceutical Biology, Albert-Ludwigs-University, Schänzlestr. 1, D-
79104 Freiburg, Germany
c Department of Internal Medicine, Division of Gastroenterology, University
Hospital Zurich, Rämistr. 100, 8091 Zürich, Switzerland
d Department of Medical Parasitology, Swiss Tropical Institute, Socinstr. 57, CH-
4002 Basel, Switzerland
e Department of Infectious and Tropical Diseases, London School of Hygiene and
Tropical Medicine, Keppel Street, London, UK
f Department of Pharmaceutical Biology, University of Basel, Totengässlein 3, CH-
4051 Basel, Switzerland
g current address: Centre for Pharmacognosy and Phytotherapy, The School of
Pharmacy. 29/39 Brunswick Sq., London WC1N 1 AX, UK
Submitted to
Journal of Ethnopharmacology
Publication III
Abstract
As part of an ethnobotanical field study bulk samples of 48 medicinal plants were
collected in three Yucatec Mayan communities. Based on their documented uses
the plants were evaluated using several biological assays.
Three species - Piscidia piscipula, Jatropha gaumeri, Casimiroa tetrameria -
employed to treat gastrointestinal disorders showed remarkable activity against
Helicobacter pylori. Crossopetalum gaumeri showed activity against Giardia
duodenalis. In the group of plants used for dermatological conditions several
species were active against gram-positive bacteria and Candida albicans.
Cytotoxic effects against KB cells were found for C. gaumeri, Diospyros anisandra,
Jatropha gaumeri, Croton reflexifolium, Dalea carthagenensis and Alvaradoa
amorphoides. D. carthagenensis and Luehea speciosa, were found to be active in
the NF-kB test. Oestrum nocturnum, applied in cases of pain and fever, showed a
weak activity against Plasmodium falciparum. Bauhinia divaricata exhibited
antihyperglycemic activity. None of the six species of the group of "women' s
medicine" showed relevant affinity to the D2 dopamine receptor.
Based on this evaluation, plants with strong activities should be further investigated
phytochemically and pharmacologically in order to better understand their potential
in the treatment of illnesses.
Keywords: Yucatec Maya, ethnopharmacological evaluation, medicinal plants,
traditional medicine, antibacterial, anti-inflammatory, antihyperglycemic,
antiparasitic, D2 receptor binding.
135
Publication III
Introduction
One of the numerous objectives of medical ethnobotany is the selection of
culturally important plant species in order to further evaluate them for
pharmacological activity (Browneret al., 1988; Etkin, 1994; Famsworth, 1988; Frei
et al., 1998a, Messer, 1978). For the evaluation of ethnopharmacopoeias,
bioassays should be chosen based on indigenous uses (Frei et al., 1998b; Heinrich
et al., 1992a, b; Lewis and Elvin-Lewis, 1994).
The knowledge of medicinal plants was a part of the ancient Maya culture and they
are still utilized by the Yucatec Mayan inhabitants on the peninsula of Yucatan,
Mexico (Roys, 1933, De Landa, 1992). During an ethnobotanical study in three
Mayan villages (Feb. 1994-June 1995; Sept. 1996-Oct. 1996), 360 medicinal plants
and 1828 reports on their uses were documented. The uses of the plants were
divided into nine therapeutical groups (Ankli et al. 1999, Heinrich et al. 1998).
Forty-eight species were chosen and evaluated in bioassays relevant to the
following groups of illnesses: gastrointestinal disorders, dermatological conditions,
women's medicines as well as pain and/or fever. Plants used to treat "diabetes"
were also tested (Table 2).
Materials and methods
Plant material
The plants were collected in the villages and surroundings of Chikindzonot, Ekpedz
and Xcocmil, Yucatan (Mexico). Authenticated voucher specimens were deposited
at the Herbarium of the Centro de Investigacion Cientffica de Yucatan (CICY) in
Mérida, the National Herbarium of Mexico (MEXU), the Instituto Nacional
Indigenista (INI) in Valladolid, Yucatan, the ETH Zurich (ZT) and the Institut für
Pharmazeutische Biologie in Freiburg, Germany (AANK1-654). They were
identified by comparison with authentic specimens and in some cases with the
assistance of specialists at CICY, MEXU and K (Kew, Royal Botanical Garden,
UK).
136
Publication III
Extract preparation
Shade-dried and powdered plant material (20g) was extracted by maceration with
100 ml dichloromethane/methanol 2:1, repeating the process 2 times during 36 h.
The filtered solvents were combined and evaporated under vacuum to give the
non-polar extract A. The residue of the dichloromethane/methanol mixture was
dissolved in 100 ml methanol/water 7: 3 and macerated 2 times during 24 h. The
solvents were combined, most of the methanol was evaporated and the resulting
water extract was partitioned between n-butanol (3 x 30-50 ml) and water. The n-
butanol fractions were evaporated to obtain the polar fraction B.
In case of S. aureus and Y. enterocolitica extract C was used. To get extract C,
plant material (10 g) was extracted once with ethanol 96% and twice with ethanol
70%.
Bioassays
Antibacterial and antifungal activity
The organisms used to test biological activities are listed in Table 1. Antimicrobial
tests were performed by the disc diffusion technique (Rios et al., 1988; DIN, 1992).
An aliquot (30 ul) of strain stock suspension was transferred into 5 ml broth
(Nutrient Broth for bacteria, Sabouraud Liquid Medium Oxoid for fungi). 10 ml of
Mueller-Hinton agar or malt extract agar (for C. albicans) was inoculated with 50 til
of overnight culture of the test organisms and poured over the agar base. Paper
discs (6 mm Blanc Discs, Oxoid) were impregnated with 200 u.g and 600 u.g plant
extracts, respectively, and placed on the inoculated agar. After 16 h incubation for
bacteria/fungi at 37 °C the plates were sprayed with MTT (methylthiazolyl-
tetrazolium chloride, Fluka). The inhibition zones were measured in mm.
Susceptibility tests with H. pylori and C. jejuni were carried out on Wilkins Chalgren
agar plates supplemented with 5% defibrinated sheep blood and the following
antibiotics: 2 ug amphotericin B/ml, 6 ug vancomycin/ml, 5 ug cefsulodin/ml, and 5
ug trimethoprim/ml. 100 ul of a thick H. pylori suspension (yield from one agar
137
Publication II
plate, resuspended in 1 ml PBS: phosphate buffered saline) were applied onto the
plates, the discs soaked with plant extracts (600 pig) and placed on the plates.
They were then incubated under a water-saturated, micro-aerophilic atmosphere at
37 °C for one day {C. jejuni), or three to five days {H. pylon) and the growth was
controlled regularly.
The MIC determination for H. pylori was carried out in a modified minimal medium
according to Nedenskov (1994). Different concentrations (in the range from 0.3 to
200 jig/ml) of the plant extracts were added to the uninoculated medium (5 ml in a
25 ml Erlenmeyer flask). A fresh H. pylori culture was used as a 1 % inoculum and
grown under micro-aerophilic conditions in a water saturated atmosphere at 37 °C
in a rotary shaker (G25; New Brunswick Scientific, New Jersey, USA). The
incubation was continued for two days at 175 rpm. After two days the optical
density of the cultures were photometrically determined at 600 nm (DU-64
spectrophotometer, Beckman, UK). The MIC value was defined as the extract
concentration not allowing visible growth (less than 0.03 in comparison with 1.2 for
the control).
Table 1. Test organisms for antibacterial and antifungal activity
Microorganism Origin Clinical picture'"
gastrointestia! problems dermatologicalconditions
Bacillus cereus ATCC 10702 diarrhea
Campylobacterjejuni*
enteritis, diarrhea
Candida albicans H29 ATCC
26790
mycosis
Escherichia coli ATCC 25922 diarrhea, dysentery infected wounds
Helicobacter pylori ATCC 43504 gastritis, peptic ulcer
Pseudomonas aeruginosa r\ \ \j\j ^oy^c infected wounds
Staphylococcus aureus ATCC 25933 diarrhea (food
intoxication)
topical infection
Staphylococcus epidermidis ATCC 12228 infection (septicemia)Yersinia enterocolitica 03 enterocolitis
* Obtained from the Department of Internal Medicine, Division of Gastroenterology, University
Hospital Zurich0(Kayseretal., 1993)
138
Table
2.Ethnomedicaldataon
plan
tsstudiedandchosen
testsystems
Family
Plantname(AANK#voucher)
Group
ofuse
Plant
Tested
for
part
Acanthaceae
Ruellianudiflora(Engelm.&Gray)
Urb.(1
15)
UR
ap
1-7,14
Annonaceae
Malmeadepressa
(Baill.)
R.
E.
Fr.(161)
UR
rt1-7,14
Apocynaceae
Tabernaemontana
amyg
dali
foli
aJacq.(190)
DER
Iv1-5
Araceae
AnthuriumschlechtendaliiKunthssp.schlechtendalii(2
43)
FEM
Iv1-5,15
Aristolochiaceae
AristolochiamaximaJacq.
(350
)Gl
,FEM
rt1-10,15
Asteraceae
VerbesinagiganteaJacq.
(288
)RES
Iv1-5
BidenssquarrosaLess.
(121
)Gl
ap
1-12
Basellaceae
Anredera
vesicariaC.FGaertner(196)
DER
Iv,tu
1-7
Bignoniaceae
Parmentieramillspaughiana
(L.)
Williams(1
35)
UR
Iv1-5,14
Parmentieraaculeata
(Kunth)Seem.
(096)
UR
Iv,
rt1-5
Bombacaceae
Pseudobombax
elli
ptic
um(Kunth)Dugand
(275
)RES
Iv1-5
Boraginaceae
Ehretia
tinifolia
L.(0
21)
RES,UR,PFE
Iv1-5,13
Bromeliaceae
Aechmea
bracteata
var.bracteataGriseb.
(167
)DER
Iv1-7
Cactaceae
Hylocereusundatus
(L.)
Britton&Rose
(427
)Gl
,UR
Iv1-12,14
Caesalpiniaceae
Bauhinia
divaricata
L.(0
07)
RES,UR,Gl
Iv1-10,14
CaesalpiniagaumeriGreenman
(155)
PFE
Iv1-7,13
Celastraceae
Crossopetalumgaumeri(L
oes.
)Lundell
(038
)Gl
,DER
Iv1-12
Cucurbitaceae
Ibervilleami
llsp
augh
ii(Cogn.)
C.Je
ffre
y(0
94)
DER
tu
1-7
Ebenaceae
DiospyrosanisandraBlake(1
34)
DER
Iv1-5
DiospyroscuneataStandi.
(341
)DER
iv1-5
Euphorbiaceae
Croton
reflexifoliusKunth
(143
)DER
Iv1-5
datrophagaumeriGreenman
(419
)Gl
,DER
rt1-12
Flacourtiaceae
CaseariacorymbosaJacq.
(150
)DER,PFE
iv1-7,13
Lamiaceae
Salviamicrantha
Desf.(0
25)
DER,FEM
ap
1-7,15
Metiaceae
Cedrelamexicana
L.(3
01)
RES
Iv1-5
Moraceae
BrosimumalicastrumSw.
(092
)RES
Iv1-5
Dorsteniaco
ntra
jerv
aL.
(330
)Gl
,FEM
rh
1-12,15
Myrtaceae
Psidiumsarîorianum(Berg)
Nied.(211
)DER,
Gl
Iv1-10
Nyctaginaceae
Neeaps
ychotrioides
F.D.Sm.
(274)
DER
Iv1-7
Pisoniaaculeata
L.(154)
FEM
Iv1-7,15
Papilionaceae
Daleacarthagenensis
var.barbata
(Oer
st.)
Barneby
(125
)DER
Iv1-5
Piscidiapi
scip
ula
(L.)
Sarg.
(123
)Gl
,RES
iv1-10
Phytolaccaceae
PhytolaccaicosandraSims
(388
)DER
fr1-5
Rivinahumiiis
L.(0
89)
DER
ap
1-5
Polygonaceae
Neomillspaughiaemarginata
S.
F.Blake(2
03)
DER,RES
Iv1-5
Polypodiaceae
Microgramma
nitida
(J.Sm.)
A.ReedSm.
(183
)Gl
wp
1-10
Rubiaceae
Rutaceae
Sapotaceae
Sela
gine
llac
eae
Simaroubaceae
Solanaceae
Sterculiaceae
Tiliaceae
Borreria
vert
icil
lata
G.Meyer
(276
)MorindayucatanensisGreenman
(113
)Casimiroa
tetrameriaMillsp.(0
49)
ChrysophyllummexlcanumBrandegee
(386
)Manilkarazapota
(L.)
Royen,Achraszapota
L.(2
34)
Sela
ginell
alo
ngis
pica
taUnderw.
(214
)AlvaradoaamorphoidesLiebm.
(136
)Cestrumnocturnum
L.(050)
SolanumerianthumG.Don
f.(3
34)
Solanumnigrum
L.(2
67)
HelicteresbaruensisJacq.
(176
)Lueheaspeciosa
Wiild.(3
47)
DER
ap
1-7
DER
fr1-7
Gl,PFE
Iv1-12
Gl
rt1-10
Gl,PFE
ba
1-10,13
UR,RES
ap
1-5
DER
Iv1-7
DER,PFE
Iv1-5,13
DER
iv1-5
DER
Iv1-5
FEM
Iv1-5,15
DER
Iv1-
7,11,12
UR:
urological
problems
(includi
ng"d
iabe
tes"
),DER:
dermatologicalconditionsin
clud
ing
inju
ries
causedbyvenomous
animals;
FEM:women's
medicines;
Gl:ga
stro
inte
stin
aldisorders;PFE:
illnessesassociated
withpa
inand/orfe
ver;
RES:
resp
irat
ory
illn
esse
s;ap:
aerialpa
rts;
ba:ba
rk;
fr:fr
uits
;Iv:leaves;
rh:rhizome;
rt:ro
ot;
tu:tuber;wh:whole
plan
t;1:
B.cereus;
2:
E.
coli;3:
C.
albicans;
4:KB-cell
line;
5:NF-kB;
6:
P.aeruginosa,
7:
S.ep
ider
midl
s;8:H.
pylori;
9:
C.jejuni;10:
G.duodenalis;
11:
S.aureus;
12:
Y.enterocolitis;
13:
P.fa
lcip
arum
;14:cx-amylase;
15:D2-receptorbindingassay
Publication III
Cytotoxicity study using KB cell culture
The cytotoxicity of the plant extracts was assessed using the KB cell line (ATCC
CCL 17; human nasopharyngeal carcinoma). The test was carried out with some
modifications according to the screening technique of Swanson and Pezzuto
(1990). The assay was performed in 96-well plates (Falcon) with an inoculum of
2.5 x 104 cells/ml. Total volume was 150 ul. The dried extracts were dissolved in
ethanol. Water was added to dilute the solution five fold. Concentration of 50 Lig/ml
with max. 1% ethanol was tested. These solutions were diluted 20 fold by mixing it
with culture medium. For active extracts, IC50 values were determined. The
quantification was performed by adding 15 ul of a solution of MTT with 5 mg/ml in
PBS (Mosmann, 1983). After incubation at 37 °C for 4 h, the metabolically active
cells produced an insoluble formazan dye. The medium was drawn off and the
formazan dye was dissolved using 150 ul of 10 % SDS (sodium dodecylsulfate) in
water. After 24 h of incubation at room temperature, the optical density was
measured at 540 nm using a microplate reader (MRX, Dynex Technologies).
Inhibitory activity on NF-kB (Nuclear Factor kB)
Anti-inflammatory activity of the plant extracts was assayed in EMSA shift
experiments (Electrophoretic Mobility Shift Assay) using the inhibition of NF-kB
binding to a radioactive labeled oligonucleotide as a molecular target. The
bioassay was carried out as described in Bork et al. (1996).
Antimalarial activity
Antimalarial activity was assessed for the chloroquine resistant K1 strain and the
chloroquine sensitive T9-96 clone of P. falciparum. The parasites were maintained
in continuous culture of infected A+ human red blood cells in RPMI 1640
supplemented with 6.9 mg/ml HEPES, 2 mg/ml glucose, 2.33 mg/ml NaHC03, 50
urg/ml hypoxanthine, 40 ug/ml gentamicin (all Sigma) and 10% A+ serum (North
London Blood Transfusion Center) (Fairlamb et al., 1985). Antimalarial IC50 values
were assessed using the modified in vitro lactate dehydrogenase assay (Makler et
141
Publication III
al., 1993). Extracts were tested in concentrations from 1000 to 4.12 u.g /ml (3-fold
dilution). Fifty jlx! of a 1% parasitemia blood suspension (predominantly ring form)
were added to 50 pi of drug solution in RPMI 1640 (final hematocrit 2%). The 96-
well microtiter plates were incubated for 48 h at 37 °C in an atmosphere of 1% 0?
and 3% C02 in balanced N2. After the incubation period, 20 ul of the parasite
suspension was added to 100 ul of Malstat reagent (Flow Incorporated, USA)
and incubated at room temperature (RT) for 15 minutes before adding 20 ul of
freshly made 1:1 NBT/PES-mixture (2 mg/ml and 0.2 mg/ml respectively) to each
well. The plates were reincubated for 20 min at RT in the dark and subsequently
read at 650 nm. All compounds were tested twice in triplicate.
Giardia duodenalis
G. duodenalis trophozoites were cultivated in Diamondi's modified TYI-S-33
medium (Keister, 1983) supplemented with 10 % heat inactivated fetal calf serum.
The in vitro assay was performed as described for the Alamar Blue® assay for
trypanosomes by Raez et al. (1997) with modifications for G. duodenalis WB strain
(isolated 1982 from a human in Afghanistan). Briefly, 200 uJ of a trophozoite
suspension were inoculated into 96-well microtiter plates (Costar, USA) at a
density of 4 x 105 trophozoites/ml culture medium. The trophozoites were
incubated in the presence of serial 3-fold dilutions of extracts for 72 hours at 37 °C.
Wells without drug served as controls. Minimum inhibitory concentration (MIC) was
determined microscopically after 70 hours of incubation (the lowest drug
concentration at which no trophozoite with normal morphology could be observed).
Ten uJ Alamar Blue® were added to each well and after 2 hours of incubation the
fluorescence determined using a fluorescence measuring instrument (Cytofluor,
Millipore; excitation wavelength at 530 nm, emission at 590 nm). IC50 values were
calculated by linear interpolation selecting values above and below the 50 % mark.
142
Publication III
Dopamin D2 receptor binding assay
Two concentrations of extracts (100 ug/ml and 10 j-tg/ml) were tested in the
dopamine receptor binding assay. The affinity of the extracts to the dopamine
receptor was assessed according to Berger (1998).
a-Amylase assay
Plant extracts (1, 3, 6 mg/ml in 5 ul solvent) were mixed with 45 ul of amylase
reagent (ET-G7 PNP 1,0 mmol/l, magnesium chloride 10 mmol/l, sodium chloride
50 mmol/l, cx-glucosidase 25.000U/I, buffer pH 7.0, sodium azide 0.05%) obtained
from Sigma Diagnostics and incubated at 37 °C for 2-10 min. The absorbance was
recorded at 405 nm versus water as a reference. The incubation was continued
and the absorbance was read after exactly 1 and 2 min. The amylase activity was
calculated according to Pierre et al. (1976).
Results
All polar and non-polar extracts of the 48 plants were screened for cytotoxic activity
against KB cell culture, B. cereus, E. coli, C. Candida and in the NF-kB test.
Extracts were additionally evaluated in selected test systems, which are of direct
relevance to the indigenous uses of the species (Table 2). The microorganisms
chosen cause gastrointestinal problems and/or dermatological illnesses, and are of
direct relevance for these conditions (Table 1). The extracts, which showed
noteworthy positive activities, are listed in Tables 3-7.
Active plants for gastrointestinal problems
Gram-positive and gram-negative bacteria as well as protozoa (G. duodenalis),
known as causes of gastrointestinal problems, were used to determine activities of
species used for gastrointestinal disorders (Table 3). Six species showed at least
some activity against G. duodenalis with IC50 values <100 ug/ml, three of them with
MIC values <100 jig/ml. The most active extract was the non-polar extract (A) of
Crossopetalum gaumeri (IC50: 2.1 ug/ml, MIC: 6.3 ug/ml), whereas the polar extract
143
Publication III
(B) showed very weak antiprotozoal activity. The non-polar and polar extracts of
Psidium sartorianum, Piscidia piscipula, Bidens squarrosa, Casimiroa tetrameria
and the non-polar fraction of Bauhinia divahcata showed a weak activity with IC50
values between 20 and 90 ug/ml. Several plants were active against H. pylori,
which is considered to play a critical role in the pathogenesis of gastritis and peptic
ulcer (Eaton et al, 1991). The non-polar extract of P. piscipula showed the highest
activity (MIC: 0.7 jig/ml) followed by the polar one of the same species (MIC: 3
(.ig/ml) (Fabry et al., 1996; Cellini et al, 1996). The non-polar extracts of Casimiroa
tetrameria (MIC: 3 ug/ml) and Jatropha gaumeri (MIC: 5 ug/ml) were active against
H. pylori. Other active extracts were Dorstenia contrajerva (A and B) and the polar
extracts of Psidium sartorianum, Microgramma nitida, Chrysophyilum mexicanum.
The non-polar root extract of J. gaumeri was found to be the most active plant
tested against gram-positive B. cereus. Extract A of C. gaumeri as well as extracts
A and B of P. sartorianum showed weak activities against this strain. The ethanol
extract of G tetrameria weakly inhibited the growth of S. aureus. There was no
significant activity against the gram-negative bacterium C. jejuni. The butanol
fraction of C. gaumeri and the dichloromethane/methanol extract of Dorstenia
contrajerva weakly inhibited the growth of this strain. None of the tested plant
species used for gastrointestinal problems showed activity against E. coli and V.
enterocolitica.
Active plants for dermatological conditions
Selected plants, documented as medicines for dermatological conditions, were
evaluated for anti-inflammatory, antibacterial and antifungal activities (Table 4). To
determine the cytotoxicity of the plant extracts, KB cell line and the HeLa cell line
were used (Table 4).
Crossopetalum gaumeri showed the most potent effect in the NF-kB test with an
inhibitory activity down to a concentration of 25 jig/ml. Already at 100 jig/ml the
polar fraction was cytotoxic to the cell line during the period of the test. The non-
144
Table
3.Evaluation
ofplantspeciesused
forga
stro
inte
stin
aldisordersbytheYucatecMaya
Ext.
KB
IC*
G.duodenalis
H.
pylori
B.cereus
S.aureus
Plantname
MIC
ICso
MIC
600ug
200ug
600ug
200ug
[ug/
ml]
[ug/
ml]
[ug/
ml]
tug/
mi]
[mm]
[mm]
[mm]
[mm]
Bidenssquarrosa
B_
-
70
nt
.-
.-
Bauhiniadivaricata
A-
-
51
nt
--
-nt
Crossopetalumgaumeri
A0.7
6.3
2.1
nt
-
12
-
B10.2
-
90
nt
--
-
nt
Jatrophagaumeri
A7.8
--
58
23
-
Dorsteniaco
ntra
jerv
aA
--
-
10
4-
--
B-
--
nt
3-
-nt
Psidiumsartorianum
A-
69.2
51
nt
-<1
1nt
B-
-
65
nt
4-
1nt
Piscidiapisc
ipul
aA
-41
27
0.7
11
--
nt
B-
-
70
312
--
nt
Microgramma
nitida
B-
--
nt
3-
-
nt
Casimiroatetrameria
A-
-72
34
--
C:1
Chrysophyllummexicanum
B-
--
nt
3-
-nt
Metronidazole
8.5
5
Ornidazole
1.1
0.5
Tetracycline
20
(10ug)
Ampicillin
23
(100ug
)Kanamycin
19(40ug
)Streptomycin
19
(50ug
)Chloramphenicol
22
(25ug)
8(10
ug)
8(10ug)
Cipr
oflo
xacin
1(0
.01)
KB:
cytotoxicity,
(-):
1C50
>50
ug/ml);
Gl:Giardiaduodenalis,
(-):
IC50>100
ug/m
l;H.
pylo
ri:
(-):<2mm;
Antibacterial
activitywasmeasuredas
inhibitionzone
in[mm];
(-):
no
activity;A:non-polar
extract,
B:po
larextract,
C:ethanol
extract;
nt:nottested.
Table
4.Evaluation
ofpl
antspeciesused
fordermatological
conditionsbytheYucatecMaya
Plantname
Ext.
KB,
IC50
NF-kB
S.epidermidis
200
ug
600ug
[ug/
ml]
[ug/
ml]
[mm]
[mm]
Aechmea
bracteata
B_
_3
5
Crossopetalumgaumeri
A0.7
25
23
B10.2
*-
-
Dios
pyro
sanisandra
A14
100
nt
nt
Diospyroscuneata
A-
75
nt
nt
Jatrophagaumeri
A B A
7,8
--
Croton
reflexifolium
39
_
nt
nt
Caseariacorymbosa
A-
-
11
Psidiumsartorianum
A-
--
1B
--
1.5
2Morindayucatanensis
A-
-
24
Daleacarthagenensis
A31
150
--
Alvaradoaamorphoides
A10
--
-
B14
--
-
Lueheaspeciosa
A-
100
--
Podophyllotoxin
0.006
Parthenolide
10uM
PDTL
100'uM
Chloramphenicol
8(1
0ug)
8(1
0ug)
Tetracycline
Miconazole
E.
coli
C.albicans
600ug
600ug
[mm]
[mm]
6(10ug)
5(1
ug)
KB:
Cyto
toxi
city
,{-
):IC
>50
ug/m
l;NF-kB(HeLa
cell
line),
(-):>150
ug/ml;*cy
toto
xicat100ug/mldu
ring
theperiod
ofthe
test
;Antimicrobial
acti
vitywasmeasuredas
inhibition
zone
in[mm];
A:non-po
larex
trac
t,B:po
larex
trac
t,nt
:not
tested.
Publication HI
polar extracts of Diospyros anisandra, D. cuneata and Jatropha gaumeri inhibited
NF-kB. These effects are probably due to the potent cytotoxicity of the extracts.
The non-polar fractions of Dalea carthagenensis and Luehea speciosa elicited
remarkable inhibition of NF-kB binding at 150 ug/ml and 100 ug/ml, respectively.
The former one showed cytotoxic activity on KB cells (IC50; 31 ug/ml), whereas no
such effect could be detected for L. speciosa.
Several extracts showed antibacterial effects against gram-positive S. epidermidls
and gram-negative E. coli. The most active extract against the former strain was
the butanol extract of Aechmea bracteata. The non-polar fruit extract of Morinda
yucatanensis was active against S. epidermidls. Other active ones were: the non-
polar fraction of C. gaumeri and Casearia corymbosa as well as both extracts of P.
sartorianum. The growth of E. coli was inhibited by the dichloromethane/methanol
extract of D. anisandra at 600 jig of the tested plant species used for
dermatological conditions showed activity against S, aureus and P. aeruginosa.
Antimalaria activities
Plants used against fever and/or pain were screened in vitro for antimalaria activity
against P. falciparum (Table 5). The non-polar extract of Oestrum nocturnum
showed some antimalaria activity without exhibiting overt cytotoxicity.
D2-receptor binding affinities
Dopamine-agonists are used in the treatment of the premenstrual syndrome (PMS)
(Steiner, 1997). For the dopamine D2 receptor binding assay five plant species of
the group of "women's medicine" were chosen (Table 2). The affinities of the
extracts on the dopamine receptor, which inhibit the secretion of prolactin, were
tested. No significant activity could be observed. The non-polar extracts of Salvia
micrantha and Aristolochia maxima showed weak receptor affinities.
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Publication III
Table 5. Anti-malaria activity of plant species used against fever and pain by the Yucatec
Mayai
Plant name Ext. IC50 against HB3 (CS) IC
50 against K1 (CR)[ug/ml] [ug/ml]
Ehretia tinifolia A
B
A
B
A
B
A
--
Caesalpinia gaumeri --
Casearia corymbosa 441.40(2.26) 494.49 (3.07)
Manilkara zapota --
Cestrum nocturnum
B
A
B
172.40(2.80) 283.31 (0.48)
Chloroquine 7.96 ng (0.03) 193.96 ng (0.81)
K1: chloroquine-resistant (CR) strain of Plasmodium falciparum;HB3: chloroquine-sensitive (CS) clone;
(- ): >500 ug/ml; no extract showed cytotoxicity in KB cell line (IC 50>50 ug/ml
Hypoglycemic effects
The polar extracts of five plant species, used against "diabetes" were tested for
their ability to inhibit a-amylase. The inhibition of the enzyme leads to a reduced
splitting of poly- and disaccharides of food in the colon. It comes to a delayed
resorption and to a balance of the value of blood sugar (Keller and Berger, 1983).
Bauhinia divaricata was the most potent inhibitor of the enzyme, followed by
Hylocereus undatus (Cactaceae) (Table 6). Polyphenols, known to interfere with
enzymes, don't occur very freguently in these genera or families (Hegnauer, 1989;
Hegnauer and Hegnauer, 1994).
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Publication III
Table 6. Anti-diabetic plants tested for a-amylase inhibition effect
Extract: Extract: Extract:
Plant name Ext. 1 mg/ml 3 mg/ml 6 mg/ml
[U/l] [U/l] [U/l]
Ruellia nudiflora B 435 403 378
Malmea depressa B 400 288 79
Parmentiera millspaughiana B 424 205 93
Hylocereus undatus B 255 0 0
Bauhinia divaricata B 0 0 0
Negative control: 620 U/l; control with methanol: 548 U/l
Other activities
Some extracts showed activities in assays which are not directly related to the
indigenous uses (Table 7). The antibiotic effects of the polar and non-polar
fractions of Caesalpinia gaumeri used for pain of the body and headache are
particularly noteworthy.
Discussion
The species evaluated in this paper were selected because of their cultural
importance with the Yucatec Maya and because of their use(s) for specific
syndromes. In this section the relevance of these findings for interpreting the
indigenous uses is discussed.
The roots of Crossopetalum gaumeri are used orally for diarrhea and snake bites
and topically to prevent inflammation after a snake bite. In a detailed
phytochemical study of C. gaumeri the non-polar fraction showed the presence of
terpenoids, whereas the butanol extract consisted of several cardenolides (Ankli et
al.; 2000, accepted). The study exhibited that the terpenoids are responsible for
antibacterial activities, which support the therapeutical value for diarrhea. The
potent antiprotozoal effect as well as the potent inhibitory activity on NF-kB is
probably due to the high non-specific cytotoxicity of the extracts.
149
Table
7.
Activities
inthebiological
assayswhichcould
notdirectly
becorrelated
totheYucatecMayanuses
Plantname
(group
ofuse)
Ext.
NF-kB
[ug/
ml]
C.albicans
S.epidermidis
E.
coli
B.cereus
600ug
200ug
600
ug
600
ug
200
ug
600ug
[mm]
[mm]
[mm]
[mm]
[mm]
[mm]
Y.enterocolitica
600
ug
mm
Caesalpiniagaumeri(PFE)
A B
Caseariacorymbosa(DER,NEU)
Casimiroatetrameria(Gl)
A
Diospyrosanisandra(DER)
A
Lueheaspinosa(DER)
C
Piscidiapisc
ipula
(Gl)
A
Verbesinagigantea(RES)
B
100
2 1
3 2
2 1 nt
3 2 5
Antibacterial
acti
vitywasmeasuredas
inhibitionzone
in[mm];
nt:nottested.
Positivecontrol:
s.Table
3-4.
Publication III
The leaves of Piscidia piscipula are applied as a medicine for gastrointestinal
disorders (especially diarrhea and cramps) and for cough. The remarkable
activities of this species against H. pylori and to some extent against G. duodenalis
may be a reason why Yucatec Maya value this plant for treating gastrointestinal
problems. In a study by Caceres et al. (1991) P. piscipula is shown to have
antimycotic effects. Another species, P. erythrina L., has been widely investigated
and isoflavones were found as spasmolytic principles (Delia Loggia et al., 1994).
Among other ethnic groups, P. piscipula is used as fish poison (Acevedo-
Rodriguez, 1990). The determined activity in the NF-kB assay is of no direct
relevance to the traditional use of this plant (Table 7).
Bauhinia divaricata is used for a variety of illnesses like gastrointestinal problems
but more frequently for "diabetes" and respiratory problems. Its activity against G.
duodenalis may be one of the reasons for the indigenous use. The leaf extract of
B. purpurea L. was reported to have significant antidiarrheal activity in vivo
(Mukherjee et al., 1998). A possible hypoglycemic activity was also found in our a-
amylase test which supports the reported hypoglycemic effects in a previous study
by Roman R. et al. (1992).
The roots of Jatropha gaumeri are used for diarrhea and the resin is used as a
medicine for herpes (labialis). The therapeutic value against diarrhea may partially
be based on the antibacterial activity against B, cereus. Different pharmacological
effects are mentioned in the literature concerning the genus Jatropha, among
others antimicrobial effects (Odebiyi, 1980). Many taxa of the Euphorbiaceae are
known to have cytotoxic phorbol esters. Cytotoxic effects of J. curcas L. using the
brine shrimp assay were described (Gupta et al., 1996).
Psidium sartorianum is employed internally (diarrhea) and externally (mostly for
measles or any kind of pimples). The antibacterial activities {H. pylori, B. cereus
and S. epidermidis) might be of interest for the antidiarrheal therapy and the use
for skin problems. To evaluate the use against measles, antiviral tests will have to
be performed. No pharmacological data for P. sartorianum are available, but P.
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Publication III
guajava L. was shown to have antibacterial effects, especially against the
enterobacterium Shigella.
In the "Ethno-Botany of the Maya" (Roys, 1976 [orig. 1931]) Casimiroa tetrameria
is mentioned as a medicine for gastrointestinal problems including diarrhea. Today
it is still used in this way but also for rheumatism and fever. The antiprotozoal effect
as well as the antibacterial activities against H. pylori and S. aureus may be of
relevance for the internal use. The antifungal activity is of no direct relevance for
the traditional use (Table 7). No pharmacological data are available, but when C.
edulis Llave and Lex. was tested against enterobacteria, no activity was detected
(Câceres et al., 1990). The antidiarrheal and antispasmodic effects are currently
investigated (Heneka et al, in preparation).
Diospyros anisandra and D. cuneata are used for dermatological problems. The
antibiotic and the antifungal activities explain the species' use in a rational way.
The NF-kB inhibiting activities is probably a result of the cytotoxic effects.
Antimicrobial activity of Diospyros lycioides Desf. (Li et al., 1998), as well as in vivo
anti-inflammatory activity of Diospyros leucomelas Poir. (Recio et al., 1995) and
antitumor promoting effects (Kapadia et al., 1997) were reported.
Various species of the genus Croton are used in the medical system of the
Yucatec Maya for dermatological problems but also for fever and respiratory
illnesses. The resin of C. reflexifolius is used for pimples in the mouth (herpes) and
eye problems. The possible occurrence of phorbol esters in this species
(Euphorbiaceae) may express the medicinal value of the use as an antiviral drug.
The application of the resin into the eyes might have severe side effects because
of the cytotoxicity.
The leaf decoction of Alvaradoa amorphoides is applied externally for skin
problems. Cytotoxic effects were found in the KB cell line, which is a common
effect in the Simaroubaceae family due to quassinoids (Phillipson, 1995).
Luehea speciosa is an effective inhibitor of the transcription factor NF-kB, which
controls genes responsible for the inflammatory responses of the body. The
Yucatec Maya value this plant for treating skin diseases and toothache, and apply
152
Publication III
the medicine in form of plasters. The inhibitory effect on NF-kB partially explains
the use of this plant. The weak antibacterial activity against Y. enterocolitica can
not substantiate the traditional use of the plant. To our knowledge no
pharmacological or phytochemical data have been published yet.
Aechmea bracteata (Bromeliaceae) and Morinda yucatanensis are applied in form
of plasters against snake bites and infected wounds and showed antibacterial
potency against S. epidermidls. This confirms the indigenous uses. M.
yucatanensis is also employed in the treatment of warts. No data on the antiviral
effect of this species are available, but many studies on Morinda spp, concerning
antitumor or antimalaria activities are available (Awe & Makinde, 1998).
Several reports on plants applied for fever are obtained, but only one of the healer
explicitly mentioned malaria. Cestrum nocturnum is applied for children with night
fever and cold bodies. Even though the drug was not mentioned specifically for
malaria by the Maya, further testing of fractions of this plant would be of interest.
Phytochemically the species is well investigated but no data have been found for
antimalarial effect (Ahmad et al. 1991),
The therapeutical group of "women's medicine" contains a lot of plants used to
relieve pain of delivery, induce labor or help "fulfill the desire for a child" (infertility).
The symptoms of PMS were never mentioned explicitly by the Mayan healers and
midwives but reports of menstrual pain are documented. A possible explanation of
the negative results of the screening may be that the bioassay is specific for PMS
and of no immediate relevance to the indigenous uses.
Conclusion
One goal of this evaluation is to better understand the use of Yucatec Mayan plant
and their healing concepts. In this paper we show some correlations between uses
of the medicinal plants and the activities obtained in the selected bioassays. Other
indigenous uses currently cannot be explained in a bioscientific manner because
the bioassays applied are not appropriate. In other cases the symbolic aspects
might be more important to the Maya. The combination of a detailed
153
Publication IN
documentation of ethnomedical uses and bioassays, which are of immediate
relevance for these uses, are of great importance and lead to a better
understanding of the ethnopharmacopoeia of the Yucatec Maya (and other
indigenous groups).
Evaluations of plants as well as the phytochemical and further pharmacological
studies are important tasks. Organizations like WHO (World Health Organization)
and TRAMIL (Central America; popular pharmacopoeia) encourage the use of
remedies that have been proven to be safe and effective. Herbal medicines are a
valuable and readily available resource for primary health care and complementary
health care systems. Hopefully this study contributes to strengthen and promote
the use of traditional medicine.
Acknowledgements
The authors wish to thank all persons who have helped in the field study and
especially the healers, midwives and the inhabitants of Chikindzonot, Ekpedz and
Xcocmil, Yucatan, for their collaboration, for their friendship and hospitality. The
botanical identification at CICY {Centro de Investigacion Cientffica de Yucatan ),
and MEXU (National Herbarium of Mexico) was performed in collaboration with the
numerous specialists of these institutions. Particularly we would like to thank Dra. I.
Olmsted, J. Granados, P. Sirna, J.C. Trejo, Dr. R. Duran of CICY as well as O.
Tellez, Dr. R. Lira, Dr. J. Villaserior and Dr. M. Sousa of MEXU. This research
owes a lot to the help of Dr. J. Heilmann (Zürich), Dr. J, Orjala (Davis), Dr. B. Frei
Haller (Zernez), Prof, Dr. W. Schaffner (Basel) and Prof. Dr. H. Rimpler (Freiburg).
Financial support by SDC (Swiss Agency for Development and Cooperation,
Berne, Switzerland) and the SANW (Swiss Academy of Natural Sciences) is
gratefully acknowledged.
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159
Plant Evaluation
6 Additional results
6.1 Antimicrobial activity
In addition to the antimicrobial tests reported in publication III, two further strains
were used to evaluate the antimicrobial activity of the 96 plant extracts, namely
Micrococcus luteus (ATCC 9341) and Mycobacterium fortuitum (isolated from a
patient). M. luteus is a gram-positive coccus whereas M. fortuitum belongs to the
gram-negative, acid fast asporogenous rods. M. fortuitum causes dermatological
problems (local abscess) or less often illnesses of the bones and junctions. The
genus Mycobacterium, which causes important human infections such as
tuberculosis and leprosy, is of high interest due to these diseases and the problem
of resistance against known antibiotics.
The antimicrobial tests were performed with all extracts mentioned in Table 2 of
publication III using the disc diffusion technique as described in the same
publication. M. fortuitum is a slow growing bacterium, thus the cultivation takes
several weeks. Because of contamination with faster growing bacteria, the results
obtained with M. fortuitum must be interpreted with great caution.
Table 6.1. Activitiy of plant extracts against M. luteus and M. fortuitum
Plant name (family) Ext. M. luteus M. fortuitum''
200 ug [mm] [mm]
Aechmea bracteata (Bromeliaceae) B - 2
Caesalpinia gaumeri (Caesalpiniaceae)
Casearia corymbosa (Flacourtiaceae)
Crossopetalum gaumeri (Celastraceae)
Jatropha gaumeri (Euphorbiaceae)
Psidium sartorianum (Myrtaceae)
Chloramphenicol 1 ug
Tetracycline 5 ug
A 1
A -
A 1
A -
B 1
B 1
- 3.5
1.5
A: non-polar extract; B: polar extract; spat|ent material; obtained from the Institute of
Medical Microbiology, University of Zurich; -: not active:
160
Plant Evaluation
The water extracts of the 48 plants (Table 2, publication III), obtained from the
liquid-liquid partition between 1-butanol and water, were also examined against
gram positive and gram-negative bacteria, the fungus C. albicans as well as for
KB-cell cytotoxicity. In two cases the water extract were active: Psidium
sartorianum was active against S. epidermidls and M. fortuitum, and Crosso¬
petalum gaumeri showed KB cell cytotoxicity.
6.2 Comparison of disk method and TLC method
The non-polar and the polar extracts of seven species were not only tested with the
disc method but also using a TLC method. By means of micropipettes the same
quantity of extract was applied directly onto a TLC plate (9x9 cm) and 10 ml of the
overnight culture was poured over it (TLC in petri plates; 10x10 cm). The test
organisms used were B. cereus, S. epidermidls and E. coli. The inhibition zones of
the extracts using the TLC method were compared with those of the disc method
(Table 6.2).
Results: The inhibition zones of the extracts obtained from the TLC method were
generally larger than the inhibition zones of the same extracts in the disc method.
The only exception was the non-polar extract (A) of C. gaumeri tested against S.
epidermidls. Here the inhibition zone obtained from the disc method is larger than
the one from the TLC method. Generally, the differences between the two methods
were more pronounced among the polar extracts (B) and less important among the
non-polar extracts (A). This phenomenon seems to be caused by the
disadvantageous diffusion of the non-polar extracts (A) from the discs into the polar
agar medium. Furthermore, the contact of the extracts with the agar is limited in the
disc method. As a consequence, the zone of inhibition in the disk method was
generally smaller than the one of the TLC method. In order to compare results
standardized methods and standard positive controls should be used. However, in
this case, the TLC technique is preferred due to decreased diffusion problem and
minimized error.
161
Plant Evaluation
Table 6.2. Comparison of inhibition zones obtained from disc - and TLC method
Plant name Ext. B. cereus S. epidermidis t. coli
TLC Disc TLC Disc TLC Disc
Caesalpinia gaumeri A 2 2 2 2 -
(Caesapliniaceae) B 3 - 3 1 -
Casearia corymbosa A 1.5 1.5 1 1 .
(Flacourtiaceae) B 2 - 2 -.
Crossopetalum gaumeri A 1 1 0.5 2 -
(Celastraceae) B 2 - 1.5 - .
Dalea carthaginensis A 2 < 1 3 - 1
(Papilionaceae) B 2 - 1 - .
Diospyros anisandra A 7 <5 15 nt 2 < 1
(Ebenaceae) B 3 - 5 nt -
Jatropha gaumeri A 3 2 4 < 1 .
(Euphorbiaceae) B - - - - -
Psidium sartorianum A 2 < 1 2 - .
(Myrtaceae) B 2 - 2 1.5 -
Chloramphenicol 1 ug - 10 2.5 - - -
Tetracycline 1 jig - - - 2 - 6 4
Quantity of extract tested: 200 ug; the inhibition zone is given in [mm]; nt = not tested; - : not active.
6.3 Protein kinase activity: Method and results
Protein kinases (PK) are enzymes that transmit phosphate to proteins. With the
induced change of conformation in proteins, they regulate different physiological
processes. Thus, they are important in the control of growth, division and
differentiation of cells. Based on today's knowledge, overexpression and lack of
control of PK activities are involved in the creation and development of tumors.
Therefore, they are targets for the development of new drugs in tumor therapy, as
well as in treatment of atherosclerosis, psoriasis and inflammatory processes.
Method: The non-polar (A) and the polar (B) extracts of 33 medicinal plants used in
the treatment of respiratory illnesses (RES), dermatological conditions (DER),
illnesses associated with pain or fever (PFE) and women's health (FEM) were
evaluated in a high throughput screening with PKs. The catalytic activity of the
162
Plant Evaluation
enzymes was measured in 96-well microtiter plates with recombinantly produced
human PKs. By means of 33P-marked phosphate, their incorporation into the
substrate was measured. The reaction volume used was 50 ul and the incubation
time was 80 minutes at 30 °C (Schachtele and Totzke, 1999). The concentration
tested of the non-polar (A) and polar (B) extracts were 1 and 10 ug/ml. The results
in Table 6.3 refer only to the lower concentration (1 ug/ml) in order to focus on the
extracts with pronounced activity. The plant family and the authors of the species
are given in Table 2 of publication III. The following kinases were employed in the
screening:
CDK4/CyclinD1 * TIE-2 » PKB (AKT)
CDK2/CyclinE ErbB2 FLT-4
EGFR Kinase IGF-R1
Results: The results are compared with those of the KB cell test and the NF-kB test
(publication III). Of the 66 tested plant fractions tested with 8 different PK, 66
positive results on the PKs were obtained (13 %) (Table 6.3). Extract B of A.
vesicaria showed 53 % specific inhibition of CDK2/CyclinE kinase. The same
extract did not show any activity in the KB cell test and the NF-kB test. However,
the non-polar extract (A) exhibited weak cytotoxic effect at 50 ppm. The polar
extracts (B) of A. bracteata var. bracteata (58 %), H. baruensis (93 %), P. piscipula
(72 %) and S. micrantha (91 %) blocked the PKB and did not show any inhibition of
other PKs. Of these four species the lipophilic extract (A) of P. piscipula were
active in the NF-kB test, down to a concentration of 100 pig/ml and the lipophilic
extract (A) of S. micrantha showed weak KB cell cytotoxicity at 50 ppm. The
hydrophilic extracts (B) of B. verticillata, C. corymbosa and E. tinifolia were active
against IGF-R1 kinase and PKB, respectively. From these plants, the non-polar
extract (A) of E. tinifolia was weakly cytotoxic in the KB cells system at 50 ppm.
163
Table
6.3.
Inhibition
ofdifferentprotei
nkinasesby
plan
textracts
at
1ug/ml
Plantname(GROUPOFUSE)
%effect
Ext.
CDK4/
CDK2/
EGFR
1GF-R1
CycIinDI
CyclinE
Kinase
TIE-2
PKB
(AKT)
FLT-4
Aechmea
bracteata
var.bracteata,
DER,FEM
Alvaradoaamorphoides,DER
Anredera
vesi
cari
a,DER
Anthunum
schlechtendaliissp.schlechtendalii,
FEM
Bauhinia
diva
rica
ta,RES
Borreria
verticiIlata,DER
Caesalpiniagaumeri,PFE
Caseariacorymbosa,DER
Croton
refl
exif
oliu
s,DER
Daleacarthaginensis,DER
Ehretia
tini
foli
a,PFE,RES
Helicteresbaruensis,FEM
Neomillspaughiaemarginata,DER
Piscidiapiscipula,RES
Psidiumsartorianum,RES
Salviamicrantha,DER,RES,FEM
B A A A A B B B B A B B
-70
-62
-57
-5£
-53
-71
-53
-68
-53
-56
-58
-55
-65
-82
-82
-86
-86
58
-51
-63
-69
54
-83
-89
-87
68
-65
-82
55
-57
-80
-56
-55
-63
-67
-80
-75
-89
-86
-58
-97
-101
-57
-96
-65
-83
-83
-80
-92
-84
-99
-98
-97
-93
-102
-72
-82
-100
-91
-80
-72
-69
-64
-54
-95
Plant Evaluation
Quite unspecific PK inhibition was caused by the polar extracts (B) of B. divaricata,
C. reflexifolius, D. carthaginensis and N. emarginata. They were active against
three to six different PKs. From B. divaricata not only the polar (B), but also the
non-polar (A) extract inhibited the PKB. Furthermore, extract A was weakly active
in the KB cell test at 50 ppm. The genus Croton of the Euphorbiaceae family is
known to have PK activity and to be cytotoxic due to phorbol esters. The
cytotoxicity of the non-polar extract (A) of C. reflexifolius (Euphorbiaceae) was
determined as IC50= 39 ug/ml. The non-polar extract of D. carthaginensis showed
cytotoxicity in the KB cell system (IC50= 31 ug/ml) as well as NF-kB activity down
to a concentration of 150 ug/ml.
The most unspecific activities in the PK screening were obtained from A.
amorphoides, A. schlechtendalii spp. schlechtendalii, C. gaumeri and P.
sartorianum. Both extracts of A. amorphoides showed PK inhibition and
additionally strong KB cytotoxicity (A: IC53= 10 ug/ml, B: IC50= 14 ug/ml). Extract A
of P. sartorianum exhibited PK inhibition as well as weak KB cytotoxicity at 50 ppm.
With respect of the search for new and specific kinase inhibitors, these four
species are less important due to the inhibition of all PKs tested.
Comparing the results of the PK tests, it is significant that polar extracts (B) were
particularly active, whereas the non-polar ones were less active or did not show
any PK inhibition. In contrast, only non-polar extracts (A) showed activity in the KB-
cell and NF-kB tests, with the exception of extracts of A. amorphoides (publication
111). However, cytotoxicity and PK activity do not necessarily correlate, this may
partially be due to the differences in the methods used. Explanation of these
phenomena presumably can be found in the different methods of the test systems.
The kinases are enzymes produced by recombinant technology and are not
incorporated into cells, whereas the KB cell test and the NF-kB test are performed
with human cell lines. To inhibit PKs located inside the cells, the compounds first
have to pass the lipophilic cell membrane. Non-polar extracts are able to pass the
membranes more easily than polar ones and, therefore, are advantageous to
165
Plant Evaluation
inhibit the PKs. Thus, particularly lipophilic extracts showed positive effect in the
cell line tests.
Another problem, in the case of crude extracts, especially in polar ones, is the rate
of false positive results. Highly purified enzyme-based targets and cell membranes
can be affected by polyphenols and saponins, respectively (O'Neill and Lewis,
1993). This is most likely reason why a higher amount of polar extracts (B) in
comparison to non-polar extracts (A) showed inhibition in the PK test system.
Most of the crude extracts tested have the capacity to inhibit PKB. Accordingly, this
kinase seems an ineffective target for providing a useful basis for specific drug
development. However, tyrosine kinase of the ErbB2 receptor was not effected by
any of the plant products. Compound or extracts that inhibit this type of PK would
probably be more interesting for further investigations.
Is there a link between the results of the PK test and traditional phytotherapy?
The initial idea was to screen the ethnobotanically pre-evaluated drugs used in the
treatment of respiratory illnesses (RES), dermatological conditions (DER) and
illnesses associated with pain or fever (PFE) for their anti-inflammatory potency.
The results of the screening showed a relative high number of active samples as
well as promising results. The ethnobotanical approach may be a reason for these
effects. Another reason may be the false positive results based on the presence of
polyphenols (e.g. P. sartorianum). It has to be pointed out, that polyphenols are
important compounds in popular phytotherapy, however they are less important in
the search for new drugs. Their ability to unspecifically inhibit enzymes, to act as
antioxidants as well as further effects, may have positive influences in the healing
process. Generally, the PK test is too specific for evaluating plant extracts and for
explaining the mechanism of healing. Therefore, further bioassays relevant for the
species use have to be conducted. In addition, more detailed phytochemical
investigation of the extracts and subsequent PK tests with isolated compounds
have to be performed. Also biopharmaceutical aspect (resorption, distribution,
metabolism) of the compounds must be incorporated into a detailed study of the
plant remedies.
166
Plant Evaluation
6.4 Other activities
Casimiroa tetrameria:
Casimiroa tetrameria Millsp. (Rutaceae), Hylocereus undatus (L.) Britton & Rose
(Cactaceae) and Bidens squarrosa Less. (Asteraceae) are used against diarrhea.
Hence, they were tested for their antisecretory activity in the USSING chamber
model (Greger et al., 1991). The crude extract of the leaves of C. tetrameria
reversed the Prostaglandine E2-induced CT-secretion significantly. Thus, this
species was chosen for further phytochemical investigations. Among others, four
fractions each containing a different polymethoxyflavone as the main compound,
showed the highest antisecretory activity. The same polymethoxyflavone
containing fractions showed significant antispasmodic activity when tested on the
isolated guinea pig ileal smooth muscle (Samuelsson, 1991). (This work was
carried out by Bilkis Heneka at the Institute für Pharmazeutische Biologie, Albert-
Ludwigs Universität Freiburg, Germany).
Malmea depressa:
The root of Malmea depressa (Baill.) R.E.Fr. [syn.: Guatteria leiophylla (Donn. Sm.)
Saff. Ex. Standi.,- Guatteria gaumeri Greenmann (Fries, 1939)] (Annonaceae) is the
most important remedy for the treatment of diabetes type II in Yucatan. Based on
ethnobotanical information, an aqueous root extract was prepared and tested on
male Whistar rats. The test medium, dissolved in physiological NaCI-solution, was
given orally to rats after an intraperitoneal injection of 50 mg/kg streptozotozin and
waiting 7 to 15 days. In addition, the test was performed with pure NaCI-solution
and glibenclamide (positive control). The application of the aqueous root extract of
M. depressa led to promising results.
Hernandez et al. (1993) studied the effect of ce-asarone, a hypolipidemic active
principle of M. depressa. Jimenez-Arellanes and Mata (1996) phytochemically
investigated this plant. They isolated five phenylpropanoids from the CHCI3 extract
of the stem bark. It might be possible that also these compounds are responsible
167
Plant Evaluation
for the hypoglycemic effect of the remedy since several similar compounds were
found to show hypoglycemic activity (Hernandez et al., 1993; Sarges et al., 1996).
6.5 Crossopetalum gaumeri -the plant species for the phytochemical study
Crossopetalum gaumeri was chosen for the detailed phytochemical study and
further biological investigations. The following points led to this selection:
its importance as a remedy among the Yucatec Maya of the study region
the healers consensus concerning the plant use and preparation
the use in a decoction against diarrhea, which is a major health problem among
Yucatec Maya
the oral use of the fresh root against snake bites
the endemic occurrence of the species on the Yucatan Peninsula
the cytotoxic and antibacterial effects detected in the in-house bioassays
4 the lack of the phytochemical investigation of C. gaumeri
the few phytochemical studies, which have been carried out with the genus
Crossopetalum
» the importance of the Celastraceae family in the search for new, effective
compounds in the field of antibiotics and cancer therapy (Kupchan et al., 1972b).
C. gaumeri was chosen for the phytochemical investigation with the aim isolating
the antibacterial and cytotoxic principles. Furthermore, the research was carried
out to clarify the empirical phytotherapy of C. gaumeri.
168
Phytochemistry
7 Celastraceae family and the genus Crossopetalum
7.1 Botanical taxonomy
Species: Crossopetalum gaumeri (Loes.) Lundell
Genus: Crossopetalum
Family: Celastraceae (Bittersweet family)
Order: Celastrales
Subclass: Rosidae
Class: Rosopsida (former; Dicotyledoneae)
Subdivision: Magnoliophytina (Angiospermae)
Division: Spermatophyta
(Frohne and Jensen, 1998)
The family Celastraceae including the Hippocrateaceae encompasses 94 genera
with approximately 1,300 species. It is a widely distributed family, which is native to
the tropics. Fewer species are native to warmer temperate regions. The species of
this family, usually glabrous with laticifers, are trees, shrubs and lianas. The chief
genera are Cassine, Celastrus, Crossopetalum, Hippocratea, Maytenus, and
Salacia (Mabberley, 1996).
The genus Crossopetalum is represented by 36 species in tropical America. They
are little trees and shrubs with indeciduous, opposite leaves. They are basically
distributed in Central America and the Antilles, only one species is reported from
South America, namely Crossopetalum rhacoma Crantz (Gentry 1993). The same
species, which is the most widely distributed one of the genus, is also native to
Florida. On the Yucatan Peninsula two species of the genus Crossopetalum are
known, namely C, gaumeri and C. rhacoma.
170
Phytochemistry
7.1.1 Crossopetalum gaumeri
C. gaumeri has different synonyms, these include Rhacoma gaumeri (Loes.)
Standi, and Myginda gaumeri Loes (Figure 7.1). The habitat of the plant is moist,
wet thickets or thin forests, mostly on limestone, higher than 200 m above sea
level. The species is widely distributed on the Yucatan Peninsula, which includes
the Mexican states of Yucatan, Campeche and Quintana Roo, as well as in Belize
and the northern part of Guatemala. The Mayan name of the plant in Yucatan is
recorded as camba-och-lob (Standley and Steyermark, 1949). Today the Maya in
the area of the place of this study call the plant viperol.
Figure 7.1. Crossopetalum gaumeri (Loes.) Lundel
171
Phytochemistry
Botanical description of C. gaumeri (Standley and Steyermark, 1949):
"A shrub, commonly about a meter high, glabrous except in the inflorescence, the
young branches green, tetragonous, sometimes sparsely hirtellous; leaves on
rather stout petioles 12 mm long or shorter, ovate to elliptic or obovate-elliptic, 5-11
cm long, 2-5.5 cm wide, obtuse to acuminate, usually bright green when dried,
coriaceous, lustrous, narrowly long-attenuate to the base, appressed-serrate, often
conspicuously so, the nerves and vines elevated and very conspicuous on both
surfaces, the veins closely reticulate; inflorescences mostly 4 cm long or shorter,
few-many-flowered, the branches stout, the pedicels filiform, much longer than the
flowers, puberulent; flowers densely puberulent or minutely pilose; calyx 1.5 mm
broad, the lobes short, rounded; petals maroon-red, orbicular, more than 1 mm
long; stamens very short; style almost none; fruit obovoid, somewhat asymmetric,
almost or fully 1 cm long, somewhat narrowed at the base."
7.2 Phytochemistry of the Celastraceae
The family of Celastraceae and the Hippocrateaceae are discussed together due to
the similarities in chemosystematics (chapter 7.5). The members of Celastraceae
family generally contain dulcit, terpenoids, polyisoprenes, polyphenols and
alkaloids (Brüning and Wagner, 1978).
7.2.1 Terpenoids (terpenes, isoprenoids)
Cardiotonic steroids (cardenolides)
Euonymus europaea L. is the best known species of Celastraceae family in Middle
Europe. The seeds of a brilliant orange (Carotinoids) are toxic due to the presence
of cardiotonic steroids and alkaloids. The cardiac glycosides of the Celastraceae
belong without exception to the cardenolide type. The different mono-, di- and
triglycosides of the species of Euonymus are generally derived from digitoxigenin
and cannogenol.
172
Phytochemistry
Digitoxigenin: R^R^R^H, R3=CH3
Cannogenol: R1=R2=R4=H. R3=CH2OH
Strophanthidin: R1= R4=H, R2=OH, R3=CHO
Antiarigenin: R1= H, R2=R,;=OH, R3=CHO
The aqueous bark extract of Lophopetalum toxicum Loher was used as arrow
poison among Philippinan indigenous people. Cardiac glycosides from the
strophanthidin and antiarigenin type were isolated from the bark, which was shown
to be cytotoxic and to have positive inotropic activity (Habermeier, 1980; Wagner et
al. 1984). Indigenous people of the Philippiness mentioned that during the
preparation of the arrow poison contact with acidic substances must be avoided
not to decrease or neutralize the effect of the poison. This may be due to the
aldehyde function on position C-10 of the aglycon, which is easily autoxidated in
water (~> oxidation to carboxyl group -» decarboxylation) (Hansel et al., 1999).
An unusual type of cardiac glycoside was isolated from seeds of Elaeodendron
glaucum Pers. by Kupchan et al. (1977). The elaeodendrosids show double bonds
on position C-4/5 and contain double binding sugars like some cardenolides of the
family Asclepiadaceae.
0\ ^O
* Elaeodendroside
O O
173
Phytochemistry
Thus, cardenolides were found in three genera (Euonymus, Lophopetalum and
Elaeondron) of the Celastraceae (Hegnauer, 1989). With the isolation of cardiac
glycosides from Crossopetalum gaumeri a fourth genus is added.
Quinone triterpene (celastroloid, quinone methide)
The orange-red pigments of the root bark of several species of the family
Celastraceae are pentacyclic triterpenes. They have a conjugated lOrt-system with
a quinone methide part of ring A and B. These compounds are characteristic
structural types for the Celastraceae. Pristimerin and celastrol are two prototypes
of these compounds. Tingenon is another member of the quinone triterpenes.
Several celastroids showed interesting activity like high cytotoxicity and
antibacterial activity.
yRi
» Pristimerin: R^COOCHg
Celastol: R, = COOH
4 Tingenon: H^ = H, C-21 : keto group
Non-quinone triterpene
Triterpenes with friedelane and oleanan skeleton are common in the Celastraceae
as in many other taxa. Friedelane triterpenes with different oxidation levels were
isolated from several species of the Celastraceae. Some of them are precursors of
the characteristic quinone methides like the maytenon acid with antileukemic
activity (Abraham et al., 1971). Besides these triterpenes, phytosterines, lupan-
and taraxeran triterpenes have been isolated from the Celastraceae.
174
Phytochemistry
* Friedelane: R1 = H
* Maytenon acid: R^ = COOH
Diterpenes and sesquiterpenes
Several diterpenes with an abietan skeleton and their derivatives are known from
members of the Celastraceae. Interesting tri-epoxide diterpenes, particularly
triptolid and tripdiolid, with antileukemic activities were isolated from Tripterygium
wilfordii Hook f. (Kupchan et a!., 1972a). Other characteristic terpenoids are
sesquiterpenes (dihydroagarofuran) and sesquiterpene-polyalkaloids. Compounds
of both of these structure types are described to have insecticidal and insectifuge
effects (Delle Monache et al., 1984).
4 Triptolid, R,= H
* Tripdiolid, R,= OH
Dihydroagarofuran
175
Phytochemistry
7.2.2 Alkaloids
A great variety of alkaloids have been found in the Celastraceae but most of them
were isolated in very small amounts. Phenylalkylamines are well known from the
fresh leaves of Catha edulis (Vahl) Endl. (Cathin = norpseudo-ephedrin). Other
isolated alkaloids are tetrahydroisochinolin and tetrahydrochinolin alkaloids,
pyrrolizidin, peptid, spermidin, xanthin and macrocyclic alkaloids. The antitumor
active maytansinoids were isolated for the first time from Maytenus species
(Kupchan etal., 1972b).
4 General structure of
maytansinoids
7.2.3 Flavonoids and other phenolic compounds
Most of the flavonoids are kaempferol and quercetin glycosides. Catechins,
particularly ourateacatechines, are characteristic of Celastraceae. Proanthocyani-
dines, anthocyanidines and tannins have been confirmed in several Celastraceae.
7.3 Phytochemistry of Crossopetalum species
The genus Crossopetalum has not been well investigated phytochemically. From
Crossopetalum tonduzii (Loes.) Lund, sesquiterpenes (Tincusi et al., 1998) and
from Crossopetalum uragoga 0. Ktze triterpenes have been isolated (Dommguez
etal. 1984).
176
Phytochemistry
7.4 Biosynthesis of terpenoids (terpenes, isoprenoids)
At present more than 22,000 natural isoprenoids are known (Zamponi et al. 1998).
They occur in all organisms, but a great variety of them were found in higher
plants, however, they are not randomly distributed over the plant kingdom. Certain
isoprenoids or combinations of them are characteristic of plant species and are
helpful in the chemosystematic. Most terpenoids are considered "secondary"
metabolites, because they lack an apparent role in the basic processes of growth
and development. Many are thought to have ecological functions, serving as
defense against herbivores and pathogens, as attractant for pollinators and fruit-
dispersing animals, or as allelopathics.
Terpenoids are biosynthesized from isoprene moieties. Squalene, or in higher
plants oxidosqualene (C30), is the biosynthetic intermediate for triterpenoids and
steroids. During phytosterol biosynthesis, it is enzymatically cyclized to
cycloartenol. After an oxidative separation of 3 methyl groups cholesterol is built,
which is an intermediate for all steroids (Figure 7.2).
Cyclization to pentacyclic triterpenes such as ß-amyrin proceeds via the chair-
chair-chair conformation of oxidosqualene. Abe et al. (1993) demonstrated that the
proton-initiated cyclization produces first the tetracyclic dammarenyl cation. The
subsequent rearrangement leads to the baccharenyl and lupenyl cationic
intermediates and to the pentacyclic oleanyl cation. Finally a series of 1,2-hydride
shifts with elimination of the H-12 proton gives ß-amyrin. Friedelane triterpenes
belong to the oleanen triterpene group and are biosynthesized by different methyl
and hydrid shifts, particularly the 4,5-methyl shift of one of the methyl group on C-4
(Brieskom, 1987). Friedelane is supposed to be the precursor of the natural
triterpene quinone methides. Itokawa et al. (1991) suggested a biogenetic route
involving the oxidative elimination of the methyl group on position 24 from
friedelane to give pristimerin-type triterpenes (Figure 7.3).
177
^ 2,3-Oxidosqualene
enzymatic cyclization
chair-chair-boat conformation
HO'
Protosteryl cation
Cycloartenol, C30 -3C
1
chair-chair-chair conformation
Tetracyclic dammarenyl cation
Y
Baccherenyl cation
Lupenyl cation
Oleanyl cation
I
HO
Cholesterol, C27
Side chain shortening
Addition of acetyl-CoA
Figure 7.2. Biosynthesis scheme of steroids and terpenoids
Cardenolides, C23
intermediate stage:
ß-Amyrin/Friedelin
Friedelane-3-on-29-ol
CH2OH
O.
HO'
AH
A
Pristimerin
o.'^r
CHO
Cangoronine A
HO/;,..
Salaspermic acid
COOH
Maytenoic acid
Figure 7.3. Proposed biogenetic pathway forcelastroloids
£OOH
fKH
COOH
Phytochemistry
Tricyclic diterpenes, particularly abietan diterpenes, are built by an enzymatic
cyclization of geranylgeraniol (C20) to a dicyclic diterpene, namely labdadien.
Abietan is then built by ring closure of labdadien to pimaran and subsequently it is
modified by methyl shifts (Hanson, 1971) (Figure 7.4).
H^
V.*H >
Geranylgeraniol
-4$f!L-_
Labdadien
\ H
Abietan Pimaran
Figure 7.4. Biosynthesis scheme of abietan diterpenes
7.5 Chemosystematic and phylogenetic relationships
Celastraceae and Hippocrateaceae contain several common compounds like
dulcit, guttapercha, proanthocyanidines, phenolcarbon acids and celastroloids.
The very close chemotaxonomical relations between these two families as well as
other criteria justify their fusion, however, no sesquiterpene polyesters and
polyesteralkaloids have been found in the Hippocrateaceae. In recent literature like
Mabberley (1996) or Frohne and Jensen (1998) only the Celastraceae are listed
whereas the Hippocrateaceae are not mentioned as a separate family. The
180
Phytochemistry
chemical similarities of the Celastraceae with other families within and beyond the
order Celastrales is shown in Figure 7 5
Figure 7.5. Chemical relationship of the Celastraceae with other families (Bruning
and Wagner, 1978)
The phytogeny of the order Celastrales is not clear. The definition of this order
varies according to different authors. The term Celastrales was first introduced by
Bentham and Hooker (1862) and was defined, among other characters, by "ovula
erecta, raphe ventralis". They included the families Celastraceae, Stack-
housiaceae, Rhamnaceae and Vitaceae. In the Engler system of Melchior (1964),
other families, particularly Aquifoliaceae and Icacinaceae, were added to
Celastrales due to some vegetative features. They were also placed in this order in
the classification of Takhtajan (1980). However, Thome (1983) excluded
Aquifoliaceae and Icacinaceae from Celastrales, placing them in Theales and
181
Phytochemistry
Cornales, respectively. Savolainen et al. (1994) suggest that the majority of the
morphological characters used to separate families of the Celastrales are
insufficient. Therefore, they carried out molecular systematic investigations with
chloroplast genes of 19 species of angiosperms. According to their results, the
order Celstrales is polyphyletic. The lineage of Celastrales should be restricted to
Celastraceae including Hippocrateaceae. The Aquifoliaceae and Icacinaceae do
not belong to this order, but form a separate lineage. Furthermore, the authors
mentioned that Euphorbiaceae (Euphorbia, Mercurialis) comprises a sister group of
the Celastrales (Euonymus, Salacia. Hippocratea). Together, they form a
monophyletic group with the Rosaceae (Rosa, Geum, Malus) and the
Rhamnaceae (Rhamnus) (compare with Figure 7.5).
Takhtajan (1980) indicated a common ancestor for the Celastrales and Santalales,
being derived from the Saxifragales. According to Frohne and Jensen (1998),
Celastrales and Santalales are listed in the same group of order and relationships
with Urticales, Saxifragales and Caryophyllidae are mentioned.
7.6 Biological activities among the Celastraceae
Many compounds isolated from the Celastraceae exhibited biological activities,
some of them showed promising pharmacological effects (Table 7,1).
In recent years, an impressive number of phytochemical studies on Trypterygium
wiifordii Hook f. (Thunder God vine) were carried out, particularly by the Takaishi
group. The plant has been used for several illnesses in Chinese folk medicine
(Table 7.2). Due to in vitro and in vivo antileukemic activity of the alcoholic extract
a detailed phytochemical investigation was made and led to the isolation of
antileukemic diterpenoid triepoxides (Kupchan et al. 1972a). The evaluation of
triptolide showed relatively potent but non-specific cytotoxicity and modest
antitumor activity (Shamon et al., 1997). In other studies the plant species was
found to show anti-inflammatory, anti-tumor, anti-immunosuppressive and anti-
AIDS activity (Shishido et al. 1994; Morota et al. 1995; Ujita et al., 1993; Chen et
182
Phytochemistry
al. 1992). Salsaspermic acid (Figure 7.3) also isolated from Thpterygium wilfordii
is reported to show promising anti-HIV activity.
The maytansinoid alkaloids (chapter 7.2.2) isolated from several species of
Celastraceae showed high cytotoxicity (KB cells: ED5010"4-10"6 ug/ml). Maytansine
was found to have the greatest potency and has been studied extensively as an
anti-tumor agent. The compound binds to tubulin and leads to arrest of cell division
in metaphase (antimitotic). Maytansine was tested in several clinical phase I and II
studies. Beside some positive clinical observations, several toxic effects were
reported, particularly neurological and gastrointestinal toxicity, and therefore, the
clinical study was stopped (Smith and Powell, 1984). In recent years the interest in
maytansinoids has increased again due to success in binding to mice-antibodies.
These carriers bring the drug to the tumor cells where they bind to receptors and
are concentrated. Thus, the toxic effects can be avoided. This effect must be
investigated further (Chari et al., 1992).
183
Table7
1Biological
activities
ofcompounds
ofCelastraceae
Activity
Responsible
compounds
Plantname
Literature
posi
tiv-
inot
rop,
cardiotonic,
cyto
toxi
c
antitumor,anti-AIDS
antitumor,antileukemic
antitumor
antitumor
antileukemic,antitumor
cytotoxic
cytotoxic
antibacterial
antimicrobial
anti
vira
l,antimicrobial
antimalana
immunosuppressive,
antiin¬
flammatory,an
tipe
roxi
dati
on
sedativ
psychoactive
insecticidal,antifeedant
cardiacglycosides
trit
erpe
nes
maytansinoides
sesquiterpenes
dulotol
celastroloide
elaedendroside
diterpeneepoxides
celastroloide
trit
erpe
nedimers
sesquiterpenes
celastroloide
celastro1
Euonymus
sp
,
LophopetalumtoxicumLoher
CelastrushindsuBenth
Maytenussp
andothers
Celastrusst
epha
notu
foli
usMakino
Maytenus
ebenifoliaReiss
Celastrussp
andothers
ElaedendronglaucumPers
Tnpterygium
wilf
ordi
iHook
f
Maytenussp
andothers
Maytenusumbellata(R
Br)Mabb
GlyptopetalumsclerocarpumLaws
Salaciakraussu(Hary
)Harv
,
Maytenussenegalensis(Lam
)Exel1
Tnpterygium
wilfordiiHook
f,others
esteralkaloids
Celastrus
sp
,Cassmesp
'phenylalkylamines
Catha
edulis(V
ahl)
End1
sesquiterpenes
Maytenus
sp
,Celastrussp
Bishayand
Kowaiejvski
1973
Habermeier1980,Kitanakaetal,1996
KuoandKuo,
1997
Kupchanetal
1971,Kuoetal
1990
Takaishietal
,1993
Shirotaetal
1998
Abraham
et
al
,1971,Itokawa
etal
,1991
Kupchan
et
al
,1977
Kupchan
et
al
,1972a
Gonzalezetal,1996
Gonzalez
et
al1992
Sotanaphun
etal
,1999
Figueiredo
et
al
,1998,E
Tahir
et
ai
1999
Li,1993
Sassaetal
,1994
Snethetal
,1963
Hofmann
e*
ai
1955
Gonzalez
et
al1997
<//j
etai,1992
Phytochemistry
7.7 Popular medicinal use
7.7.1 Yucatec Maya medicinal use of C. gaumeri
According to various h-men (shamans), healers and midwives of the villages of
Chikindzonot, Ekpedz and Xcocmil (Yucatan) a two-cm piece of root is used as a
treatment for snake bites. It is chewed as soon as possible after the bite. The
powdered root mixed with water is put on the wound in the form of a plaster.
Sometimes the decoction is combined with other medicinal plants like Morinda
yucatanensis Greenman, Samolus ebracteatus Kunth and Anredera vesicaria C. F.
Gaertner. The rootstock of latter plant which is rich in polysaccharide is responsible
for the galenic form of the plaster. Diarrhea is a further usage of C. gaumeri. A
decoction of the root, mixed with other species, is orally used against diarrhea.
In the study region C. gaumeri is named viperol. Formerly it was called camba-och-
lob (Standley and Steyermark, 1949). The species is also listed in the Diccionario
Maya as kambaochdob, but no medicinal use is mentioned (Barrera et al., 1991).
The change of the plant name seems to come from the medicine named viperol,
which in the past was produced from the roots of C. gaumeri. According to oral
history, the medicine in the form of ampoules was used orally as an antidote
against venomous snake bites. The use of the medicine, under the name viperol, is
still known but detailed information about ingredients, preparation and place of
production could not be obtained (oral information from healers, employee of the
Instituto Nacional Indigenista and other inhabitants of the Yucatan Peninsula).
According to the healers of the study region two other species named viperol are
known. These are Urechites andrieuxii Muell. Arg. and Echites yucatanensis Millsp.
ex Standley. Both species are members of the Apocynaceae family and are also
used against snake bites.
185
Phytochemistry
7.7.2 Medicinal application of other Crossopetalum species
The Huastec Maya, situated in the northeastern part of Mexico, use
Crossopetalum uragoga O. Ktze not only as an anti-diarrheal medicine, but
also for burning eyes, hemorrhage and as a dermatological medicine
(Dominguez, 1984; Alcorn, 1984).
4 Crossopetalum parvifolium (Hemsley) Lundell is applied for dysentery among
the Tzotzil of Zinacantan in Chiapas, Mexico (Breedlove and Laughlin, 1993).
In the West Indies and Cuba, the roots of Crossopetalum rhacoma Crantz are
used in the treatment of kidney stones and the leaves are applied as diuretic
and for kidney inflammation (Morton, 1981 ).
Two species from Panama, namely Crossopetalum tonduzii (Loes.) Lund,
and C. lobatum Lundell were screened for antimicrobial and cytotoxic activity.
Both showed antibacterial activity and C. lobatum exhibited cytotoxic activity
against HeLa cells (Gonzalez et a!., 1994). Unfortunately no data about local
medicinal use of these species could be found.
7.7.3 Global medicinal use of Celastraceae species
The Celastraceae are widespread throughout the world and have a long history of
use at the folk level both for medicinal and agricultural purposes. Some of the
medicinal uses are listed in Table 7.2.
186
Table 7 2 Medicinal use of Celastraceae species
Plant species Uses (plant part used)' Source Location
* Hippocratea
- cclastroides Kunth
- excelsa Kunth
Maytenus
- amazonica C
Martius
- buxifolia (A. Rich)
Griseb
- laevis Reissek
- senegalensis
(Lamarck) Exell
* Celastrus
- angulatus Max
-scandens
Euonymus
- atropurpurea Jacq
- europaea L
tranquilizer (p), Morton 1981 Yucatan
asthma, bronchitis, cough (i) Ankli see appendix Yucatan
rheumatism flu, gastrointestinal diseases, Peru
antitumoral, (various parts), Chavez et al 1998
Stomach ache, diarrhea colds fever, menstrual Brhamas
hemorrhage (I) Morton 1981
painkiller rheumatism stimulant d'uretic(b), Western
Schultes and Rauffauf, 1990 Amazonia
arrow poison (b) gastrointestinal problems Africa,
fever, yellow fever snake bite (r) Neuwinger, Tanzania
1996, Gessler 1995
insecticide, Wang et al,1997 China
diarrhea, sores to regulate menstruation, (I), New Jersey
diuretic, laxative cancer cough (r) Still, 1998
laxative (p), eye bath, uterine problems (b) New Jersey
Still, 1998
cardiotonic, emetic purgative, insecticide and Europe
vermin, Bishay and Kowalewski 1973
Cassine matabelica skin cancer, diarrhea, stimulant (r), Bruning, Afrca, India
(Loes)Steedm 1978
bilharziosis, dysentery (r), Figueiredo et al, Mozambique
1998
dermatitis, rheumatoid arthritis, nephritis China
insecticide (r) stimulant others Chen et al
1992, Shishido etal,1994
* Salacia kraussn
(Harv ) Harv.
Tnpterygium sp
(I) leaf, (r) root, (b) bark, (p) whole plant
187
Phytochemistry
8 Methods (isolation procedure)
8.1 Thin layer chromatography (TLC)
Thin layer chromatography (TLC) was used to optimize the mobile phase for VLC,
MPLC, HPLC and open-column chromatography. The eluents were optimized
based on the eight proposed solvent groups of the PRISMA model and with
modified mobile phases for special structure types reported in literature (Nyiredy
et al., 1988; Wagner et al., 1983). TLC was further employed for chromatographic
examinations of fractions and extracts (two-dimensional TLC), for preparative
chromatography, for monitoring chemical derivation (hydrolysis) and for locating
biological activity on TLC plates. To enhance good separation, the TLC mobile
phase was transferred to off-line systems by reducing the solvent strength
(Kowalska, 1996).
8.2 Vacuum liquid chromatography (VLC)
Vacuum liquid chromatography (VLC) was initially used as separation procedure
of the crude extracts. Furthermore some small fractions were separated by RP-
VLC. The columns were dry-packed under vacuum. All extracts were adsorbed on
Celite 535 and applied to the column in order to avoid obstruction. The fractions
were dissolved in the starting mobile phase and the separation was forced by a
water suction pump producing a vacuum (Hostettmann et al., 1998).
8.3 Middle pressure liquid chromatography (MPLC)
Medium pressure liquid chromatography (MPLC) was employed to separate the
complex fractions after the application of the VLC technique. The column and
pre-column were slurry-packed under pressure (21 bar, flow speed: 8 ml/min).
After packing, the pre-column was removed. The sample (1.2 g), dissolved in a
little solvent (< 5 ml), was injected via a sample loop (Hostettmann et al., 1998).
188
Phytochemistry
8.4 High pressure liquid chromatography (HPLC)
The semipreparative high-pressure liquid chromatography (HPLC) was used as
one of the final steps of the isolation procedure. Samples were applied in the
amount of 5 mg for one injection. Further parameters see publication III. For semi¬
preparative HPLC 1-100 mg probe can be applied, depending on the column size
and particle size of the mobile phase (Unger, 1995).
8.5 Open column chromatography
The classical open column chromatography was applied for the separation of
complex fractions and for small fractions. The size of the column was chosen
according to the quantity of the sample (sample:stationary phase = approx. 1:100)
(m:m). The columns were slurry packed either with normal phase material (silica
gel) or reversed phase material (octadecyl phase) (Müller and Keese, 1988).
8.6 Liquid-liquid partition (LLP)
Liquid-liquid partition was used tor the first separation step of the methanolic
crude extract. The separation is based on the dipole interaction properties of the
solvents. This separation technique takes place under mild conditions and
without any loss of material (Müller and Keese, 1988).
189
Phytochemistry
9 Methods of structure elucidation
Several spectroscopic and spectrometric instruments exist for the process of
structure determination of natural products. The combination of physical data e.g.
UV- spectroscopy, optical rotation, MS-spectrometry and NMR-spectroscopy,
together with information of chemical methods is used to determine the structure
of compounds.
Isolate
1H NMR
13C NMR
DEPT
Inverse gated
number of carbons and hydrogens
functional groups
reliable 13C integrals
M S molecular weight
UV chromophore
Molecular formula; double bond equivalent functionalities
HSQC
DQF-COSY
TOCSY
HMBC
T INADEQUATE
one-bond correlation (C-H)
vicinal protons (CH-CH) and geminal protons (CH2)
protons in a spin system
long-range correlation (C-C-C-H), (C-C-H)
C-C coupling
Planar structure
X-Ray n
ROESY
[a]D f relative/absolute stereochemistry, optical rotation
y dérivât.J
Complete structure with relative/absolute stereochemistry
Figure 9.1. Scheme of methods for structure elucidation
190
Phytochemistry
9.1 Nuclear magnetic resonance spectroscopy (NMR)
Nuclear magnetic resonance spectroscopy (NMR) is a method which is applied
for nuclei that have a nonzero spin quantum number (e.g. 1H, 13C, 15N). When
these types of nuclei are brought in a static, homogenous magnetic field, they
interact with it. Depending on the surrounding electron density, the observed
nuclei are able to absorb energy, when they are irradiated with the appropriate
radio frequency.
The pulsed Fourier transform NMR spectroscopy (FT-NMR) is used as the
particular NMR technique in which all of the nuclei of one isotope are activated
simultaneously by an appropriate pulse. The absorbed energy is subsequently
lost to the surroundings or to other nuclei over a period of time (relaxation time).
The response of the system is monitored as interferogram (intensity as a function
of time). The resulting emission signal from the excited nuclei is known as the free
induction decay (FID) and the Fourier transformation of this decay yields the NMR
spectrum.
One dimensional NMR methods
The frequency, in which a nucleus is able to absorb energy, depends on the
environment of the particular nucleus and is proportional to the strength of the
magnetic field. It is expressed as the chemical shift ô and given in parts per
million. Functional groups are identified by characteristic 1H and 13C chemical shift
values. The number of signals gives information about the number of nuclei of the
molecule under investigation. Furthermore the multiplicity and the coupling
pattern of a signal is important. Many signals are split into several lines due to the
effect of neighboring nuclei. The frequency differences between such multiple
lines are given as coupling constants J (Hz) and depend on the number and
nature of bonds and the angular relationships of the coupled nuclei (Byrne, 1993;
Kalinowski et al., 1984).
By means of DEPT (Distortionless Enhancement by Polarisation Transfer), a
further one-dimensional experiment, the number of hydrogens bonded to the
191
Phytochemistry
particular carbon atoms can be determined. DEPT 135 distinguishes positive
signals for CH3 and CH and negative signals for CH2 resonances.
The inverse gated experiment is used as a method to get reliable integrals of
13C spectra. In order to obtain comparable integrals, it is necessary to avoid
building up of NOEs (Nuclear Overhauser Effect) in the sample; it is usually
reached by making them all equal to zero. This can be achieved by inverse
gated decoupling in which the decoupler is on during pulse and acquisition,
but is off during a relaxation delay. At the moment of pulse, none of the carbon
signals is enhanced and all have the same integrals (Sanders and Hunter,
1988). The different relaxation times, which also leads to unreliable integrals
can be avoided by increasing the relaxation delay, so that nuclei with long
relaxation times can turn back to the basic state. The inverse gated
experiment was used for detecting doubled intensities of carbons of the
symmetrical unit of compound 1 (ourateacatechin).
Two dimensional NMR methods
Two dimensional (2D) NMR techniques give more information on the interaction
between nuclei. The homonuclear and heteronuclear scalar coupling techniques
provide the direct connectivities between the atoms and enable the configuration
of a molecule to be determined (DQF-COSY, TOCSY, HMQC, HMBC,
INADEQUATE). The homonuclear dipolar coupling techniques give the
internuclear distances. Thus, the relative stereochemistry of the substance can be
determined (ROESY).
Homonuclear 2D NMR methods
The DOF-COSY (Double Quantum Filter Correlation SpectroscopY)
experiment is the standard 2D experiment for homonuclear shift correlation of
vicinal and geminal protons.
The TOCSY (TOtal Correlation SpectroscopY) experiment is employed to
define all protons within a spin-system.
192
Phytochemistry
* The ROESY (Rotating frame nuclear Overhauser Effect SpectroscopY)
experiment helps identifying pairs of protons close enough to interact through
space.
* The INADEQUATE (Incredible Natural Abundance Double Quantum Transfer
Experiment) detects signals due to 13C,13C coupling. It is used to trace the
entire C skeleton of a molecule. The experiment shows extremely low
sensitivity, therefore a high amount of material is used depending on the
molecular weight (~ 1/5 of M/1000 ).
Heteronuclear 2D NMR methods
* The HSQC (Heteronuclear Single Quantum Correlation) experiment is
employed for one-bond heteronuclear chemical shift correlation.
The HMBC (Heteronuclear Multiple Bond Correlation) is a method for
determining connectivities between 1H and 13C atoms, separated by two or
three bonds.
9.2 Mass spectrometry (MS)
Mass spectrometry involves the generation of positive or negative ions from
organic molecules that are in gas-phase. Subsequent separation and detection of
fragment ions provide information about the molecular formula and also about
structure elements of a compound (Baldwin, 1995).
Electron impact (El) MS: In this standard method, the vaporized sample
molecules are bombarded with electrons emitted from a heated filament (70
eV). Due to the energy, electrons are ejected with the formation of positive
ions.
Fast atom bombardment (FAB) MS: The compound is suspended in a viscous
liquid with low volatility (3-nitrobenzylalcohol) and bombarded by fast atoms
193
Phytochemistry
(argon or xenon) or ions. Molecular information yielded [X+Hj+ ions or [X+Na]+,
[X+K]+.
Electronspray ionization (ESI) MS: The involatile sample is injected in a
flowing liquid stream of methanol and ammonium acetate in order to achieve
better ionization ([X+H]+, [X+NHJ/, [X+Na]+; apparatus: Finnigan 7000 TSQ).
9.3 UV-Spectroscopy (UV)
Due to the electronic structure of a molecule, the characteristic absorption of a
chromophore enables the identification and classification of structural elements.
UV- Spectroscopy normally refers to the absorption in the ultraviolet (200-380
nm) and visible (380-800 nm) light.
9.4 Optical rotation
A compound is optically active, if it is able to change the direction of the linearly
polarized light. The specific rotation [ct]D, measurable angle to the plane of the
incident light, is a characteristic property of chiral compounds using the sodium D-
line (589.5 nm).
9.5 Acidic hydrolysis
In order to identify monosaccharides of sugar chains, acidic hydrolysis was
performed. Subsequent analysis of the sugar moiety by TLC in comparison with
defined sugar references allowed identifying the monosaccharides.
194
Phytochemistry
10 Plant extraction
10.1 Small scale plant extraction
Air-dried and powdered roots of Crossopetalum gaumeri (20 g) were macerated
in 70 ml petroleum ether for 4 hrs at room temperature. The subsequent extraction
was carried out in a percolator (2.5 x 30 cm) with a series of solvents of increasing
polarity, namely, petroleum ether, dichloromethane, methanol, a mixture of
methanol-water 1:1 and water. The percolation took place over a 43 h period, i.e
over 4 (petroleum ether), 8 (CH2C!2), 4 (MeOH), 18 (MeOH-H20) and 9 hrs (H20),
respectively. Chromatographic examinations of the five extracts by TLC showed
the similarity of the petroleum ether and the dichloromethane extract. Both
extracts exhibited NF-kB activity and antibacterial potency against different gram-
positive bacteria, namely, B. cereus, S. epidermidls and M. luteus. As a result of
these preliminary experiments, the process for the main extraction was
accordingly modified. Instead of petroleum ether and dichloromethane, only the
latter one was used for the non-polar extraction.
All five extracts exhibited cytotoxic activities down to a concentration of 10 ppm
based on KB cell culture. The extracts showed neither antifungal activity nor any
growth inhibition of E. coli.
10.2 Large scale plant extraction
Powdered roots (sieve 6 mm, then 1 mm) of C. gaumeri (2.26 kg) were
successively extracted in a percolator (18 x 45 cm) with distilled dichloromethane,
dichloromethane/methanol, methanol, 70% aqueous methanol mixture and de-
ionized water. From time to time the drug was manually stirred with a spatula (I =
60 cm). The CH2CI2 extraction took place over a 168 h period, the CHgCL/MeOH
and the MeOH extraction over a 120 h period. The MeOH/H20 and the H20
extractions went on for 72 h, each. The change in the polarity of the solvents was
indicated by the change in the color of the solvents. By removing the solvents in
195
Phytochemistry
vacuo, five extracts were obtained and stored in a freezer at -20°C. The
extraction scheme is shown in Figure 10.1.
Roots of Crossopetalum gaumeri
(2.26 kg)
Percolation at ambient temperature
Evaporation in vacuo
Extract: CH2CI2 CH2CI2/MeOHMeOH MeOH/H20 H20
Quantity: 94 g 23 g95 g 35 g
27 g
Color: brown-red red-orange brown red-orange light-brownI J v J v~.„ ^s
Figure 10.1. Extraction scheme
The cytotoxic potential of the CH2CI2, MeOH and H20 extracts were assessed
based on KB cell culture. The MeOH extract showed activity down to 1.25 ppm,
the brown-red CH2CI2 extract was still active at 2.5 ppm and the water fraction at 5
ppm. Furthermore the CH2CI2 extract showed antibacterial activity {B. cereus, S.
epidermidis and M. luteus).
10.3 Fractionation of the methanol extract
As a crude separation of the methanol extract, liquid-liquid partition was carried
out twice for each phase. Further fractionation was done using VLC, open column
chromatography and HPLC. UV-spectra of relatively pure fractions were obtained
on a spectrophotometer in order to optimize the wavelengths for the UV-detection
and HPLC separation of the compounds. Thus, the isolated cardenolides were
detected by X = 226 nm. The fractions obtained were collected and combined
196
Phytochemistry
The detailed isolation procedure is shown in Figure 10.2 and described in
publication IV. The numbers of the isolates in this thesis do not correspond to the
numbering system in the publication.
10.4 Fractionation of the dichloromethane extract
For preliminary fractionation, the CH2CI2 extract was separated by VLC on silica
gel. The fractions obtained were then chromatographed using MPLC and RP-
VLC. The final purification steps were carried out with semi-preparative RP18-
HPLC, C18 cartridges (Sep-Pak®) or by liquid-liquid partition. The isolation
process was guided by the antibiotic activity against S.epidermidis and with the
help of 1H NMR spectroscopy. Detailed fractionation and isolation is shown in the
flow chart (Figure 10.3) and discussed in publication IV.
197
Methanolextract(42g)
Liquid-liquid
part
itio
n60%aqueousCH3OH-CHCI3
1:1
CH3OH/H20
extract
Liquid-liquid
partition
H20-n-BuOH
1:1
CHCI3
extract
(3g)
H20
extract
(17
g)
n-BuOH
extract
(22
g)VLC
silicagel
CHCI3
-
>CH3OH
stepgr
adie
nt
Fr.9
130mg
Fr.2(24mg),
HPLCRP18
ACN-H20
20:80
Fr.5(155
mg),
HPLCRP18
ACN-H20
20:80
Fr.13-15
(6.9
g),
Opencolumnchromatographyon
silicage
lCHCl3-CH3OH-H?0
->89:10:1
->45:50:5
Fr.7
(72mg),
HPLCRP18
ACN-H20
20:80
Fr.9(250mg).
HPLCRP18
ACN-HpO
15:85
Fr.11
(640mg),
HPLCRP18
ACN-H20
15:85
5
39mg
6
2.4mgJ
Figure
10.2.
Isolationscheme
ofthemethanol
extract
Dichloromethaneextract(33
g)
VLC
silicage
ln-hexane—>EtOAc
Fr.8
7
1.8g
Fr.3
Fr.6(63mg)
Liqu
id-l
iqui
dpartition
8
14mg
Fr.11&
12
(3.6
g)MPLC
silicage
ln-hexane-EtOAc-CH3OH
15:3:0.5
10:10:5
Fr.8
(1.3
g)VLCRP18
CH3OH-H20
6:4
Fr.13(11mg)
RP18
CartridgeSe
p-Pa
k®CH3OH-H20
1:1
->CH3OH
CH3OH
Fr.24&25
(147mg)
VLC
silicage
ln-hexane-»CHCI3
Fr.10-15(3
4mg)
HPLCRP18
ACN-H20
9:1
9
5.3mg
10
2.1mg
Figure
10.3.
Isolationscheme
ofthedichloromethaneextract
Phytochemistry
11 Structure elucidation of the isolated compounds
In this chapter, additional information on the isolated compounds is discussed in
publication IV. Interesting data of the isolates are compared for each group of
natural compounds. The discussion of the isolates follows the numbers as shown
in the isolation protocols (Figure 10.2 and 10.3).
11.1 Cardenolides
Five cardenolides were isolated from the methanol extract. Four different types of
aglycones (securigenin, sarmentosigenin, sarmentogenin, and 19-
hydroxysarmentogenin) and three different types of sugar moieties were
identified, namely ß-6-deoxygulose, ß-6-deoxyallose and a-allose.
\—Q
HO—/ V"*-
HO tDH
RiO
B
HOH2C
rilOl:b-OH HO OH
Name of the isolated cardiac glycoside R1 R2 R3
Securigenin-3ß-0-ß-6-deoxyguloside (2)
Sarmentosigenin-3ß-0-ß-6-deoxyguloside (3)
19-Hydroxy-sarmentogenin-3ß-0-ß-6-deoxyguloside (4)
Sarmentogenin-3ß-0-[a-allosyl-(1 -^4)-ß-6-deoxyallose] (5)
Securigenin-3ß-0-[cc-aIlosyl-(1 -^4)-ß-6-deoxyallose] (6)
A H CHO
A OH CHO
A H CH2OH
B H CH3
B H CHO
200
Phytochemistry
Color reaction on planar chromatography (TLC)
The cardiac glycosides showed characteristic color reactions on TLC after
detection with vanillin-H2S04 reagent and heated to 110 °C for 5-10 minutes. The
color pattern included greenish to intense blue colors, depending on the
substitution pattern of the genin and the sugar moieties. Compound 5 with the
methyl group on C-10 resulted in an intensely blue spot. This phenomenon
supports the assumption that only cardenolides. which are not oxidized on C-19,
respond with a blue color (Pauli and Junior, 1990).
(1 ) orange spot; ourateacatechin
(2) greenish spot, fluorescence under UV-366 nm
(3) green-yellow spot, fluorescence under UV-366 nm
(4) green spot, fluorescence under UV-366 nm
(5) blue-green spot, fluorescence under UV-366 nm
(6) brown-greenish spot, fluorescence under UV-
366 nm
Figure 11.1. Thin layer chromatography (TLC) of the isolated compounds 1-6.
Mobile phase: ethyl acetate-methanol-water 81:11:8; stationary phase: silica gel
60 F254; detection: sprayed with vanillin-H2S04 reagent and heated to 110 °C for
5-10 min, evaluation under visible and under UV light 366 nm.
201
Phytochemistry
The vanillin-H2S04 reagent reacts with the aglycone and the sugar moiety (Stahl
and Glatz, 1982). The color reaction under visible light depends partly on the
sugar moiety while fluorescence on UV 366 nm is attributed to the aglycones
(Pauli, 1993). The transformation with Kedde reagent (NaOH, dinitrobenzoic
acid), which has not been used for the detection in this study, takes place on the
butenolide ring and formation of a Meisenheimer-complex occur (Jork et al.,
1993).
1H NMR comparison of the cardenolides
The cardenolides showed complex 1H NMR spectra due to the excessive signal
overlap in the 1.0-2.5 ppm chemical shift area. The signals at 5 4.91-4.92 {dd, J =
18.2-18.5, 1.5, H?-21) and 5 5.90-5.94 (s, H-22) are characteristic signals for the
butenolide ring of the cardenolides. The sugar moieties showed shift ranges
between 5 3.28 and 5 4.74 with a methyl group between 51.21-1.29 {d, J- 6.1-
6.7, H-6') each. The aldehyde group on C-19 of compound 3 at 5 9.97 is shifted
downfield compared to those of 2 and 5 (5 9.41 and 9.42). This phenomenon is
caused by the hydroxyl group at position C-5 of compound 3. The genin of
compound 4 is characterized by an alcohol function at C-19, replacing the
aldehyde function of 2, 3, 6 (s. 9.41-9.97). The two doublets were shifted to
higher fields and resonated at 5 3.86 and 3.71 (each d, J = 10.8 Hz). The
chemical shift of H-11 (5 3.72-4.42) changed due to different substituents at C-5
and C-19. Further characteristic signals among the five cardiac glycosides are the
triplet signals for H-17 at 5 2.93 (r, J = 6.9-7.1 Hz). The signals of H2-21 are not
clearly expressed because of water suppression at approx. 4.80 ppm. The MeOD
peak at 5 3.31 was always accompanied by a MeOH peak at lower field (Figure
11.3).
202
Phytochemistry
13C NMR comparison of the cardenolides
Two quaternary carbon signals in the range of 5 175.7-177.7 showed different
intensities due to different relaxation times. The weaker signal in higher field was
assigned to the carbonyl carbon C-20 of the butenolide ring. Due to HMBC
correlations the stronger signal was assigned to C-23. Different substituents on C-
5 and C-19 indicated differences in the 13C NMR shifts between the isolated
cardenolides. Thus, the vicinal carbons C-4 (5 29.1-34.0), C-6 (5 27.7-37,1) C-10
(5 37.5-55.1) and also C-1 (5 19.7-33.8) shifted downfield or upfield. The C-4' (5
84.0-84.1) and C-6' (5 18.2) signals of the disaccharides 5 and 6 in comparison
with the monosaccharides 2, 3 and 4 (C-4!: 72.2-73.7; C-6': 14.9-16.4) are shifted
to lower field due to the O-linkage of the second sugar moiety (Figure 11.4),
COSY and HMBC correlations of the cardenolides
The assignment of the signals of the cardenolides and their sugar moieties was
verified using COSY, TOCSY, HSQC and HMBC measurements, particularly.
With the aid of COSY and TOCSY spectra the saccharides were identified (Figure
11.5, 11.6).
Relative and absolute stereochemistry of the cardenolides
The relative stereochemistry was elucidated by ROESY experiments, the 1H,1H
coupling constants and the application of steric arguments. The results were
verified by comparing the data with those of the literature.
A/B ring junction:
Due to the 1H,1H ROESY correlations between H-5 and H-19 A/B eis
configurations were suggested for the aglycones (Figure 10.2). The comparison
with the 13C NMR data for C-5, C-1, C-7 and C-9 of all five compounds with those
of several A/B eis configuration and also with uzarigenin (A/B trans system)
verified the geometry of the A/B ring junction as eis (Drakenberg, 1990; Cheung et
al., 1981). The chemical shifts of C-5 of uzarigenin (H-a) are in lower field
203
Phytochemistry
compared to the same carbon atoms in A/B c/s-fused systems. These differences
are due to steric compression shifts induced in the latter systems by forcing the
ring system into the c/s-configuration (Wehrli, 1979).
Figure 11.2. ROESY correlations of compound 6 (600 MHz, MeOD)
* B/C ring junction:
The B/C frans-fusion was indicated by the observation of correlations in the
ROESY spectrum between H-8/H-19 (2, 3, 4, 5), H-11/H-19 (2, 5, 6), H-8/H-11
(5, 6) and the lack of any correlations between H-8/H-9 (Figurel 1.2).
C/D ring junction and H-17 orientation:
The C/D ring junction was assigned as eis, with the butenolide ring attached at
the C-17ß position. In support of this configuration, the protons of H3-18ß gave
rise to strong synaxial NOE correlations with the proton of H-11ß and H-8ß, H2-21
and H-22 of the free rotating butenolide ring showed correlations to the H3-18ß,
which indicated a ß position of the lactone ring. In addition, the chemical shift of
C-12 of the aglycones lie in the range of 48.5-50.6 ppm, whereas a butenolide
ring in C-17a position is characterized by an upfield shift of approx. 8 ppm to
approx. 5 42 for C-12 (Kawaguchi et al., 1993). Due to this evidence and in
204
Phytochemistry
comparison with the 13C and 1H shifts of digitoxigenin, C/D ring junction as eis and
C-17ß orientation were verified (Drakenberg, 1990).
Relative stereochemistry of the saccharides and H-3 orientation:
The relative stereochemistry of the sugar moieties is described in detail in
publication IV.
Mass spectrometry of the cardenolides
For the cardiac glycosides positive FAB-MS were measured due to the soft
ionization method of FAB technique. A fragment of m/z 146 indicates the
presence of a deoxyhexose (2, 3), m/z 162 substantiates the presence of a
hexose (5, 6) (Figure 11.7). For the disaccharides, 5 and 6, no molecular peaks
were obtained. Instead, peaks were obtained which indicated the loss of a
hexose [M+Na-sugar]+, [M+K-sugar]\ Based on an ESI-MS the molecular ion of
the disaccharide cardenolides was detected.
205
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11.4.13CNMR
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Figure
11.5.1H1HDQF-COSYspectrum
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11.7
PositiveFABmassspectrum
ofcompound2
Phytochemistry
11.2 Ourateacatechin
(-)-4'-0-methyl-epigallocatechin (Drewes and Mashimbye, 1993), 3,3',5,5',7-
pentahydroxy-4'-methoxy-2,3-c/"s-flavane (Weeratunga et al., 1985)
OCH,
2 ^
'%
OH
1
M,: 320
C16Hl607
Ourateacatechin (1) was isolated as a brown-orange amorphous solid. The 13C
NMR and the DEPT experiment detected 14 carbons. In the 1H NMR a double
intensity of the aromatic H-2' {6 6.55) was observed (Figure 11.8). The inverse-
gated 13C (1H) NMR experiment verified the double intensity of the methine group
at 107.12 ppm and showed the same phenomena for the quaternary carbon at
151.15 ppm (Figure 11.9). This indicated some type of symmetry in the molecule
leading to at least eight quaternary carbons, six methine groups, one methylene
and one methyl group. The combination of these results with the EI-MS, which
showed a pseudo-molecular peak at m/z 322.0 [M+2H]+, allowed the
establishment of the molecular formula C16H160T. Furthermore, the mass spectrum
revealed a pair of important fragment peaks at m/z 183 and 140 that are typical for
retro-Diels-Alder fragmentations (Garcia et al., 1993) (Figure 11.10). The 13C-
INADEQUATE NMR allowed the formulation of the carbon skeleton C4-C3-C2-
CT-C2'-C3'-C4' and C-4 to C-10 (Figure 11.11). The two fragments were
composed with the help of HMBC experiments, especially with the correlation
between H-2 and C-9 and the correlation of H-6 to C-5/8/10 and H-8 to C-6/7/10.
Thus, 4'-0~methyl-epigallocatechin (1) was identified. The stereochemistry was
213
Phytochemistry
elucidated by ROESY experiments and coupling constants. The intensive NOE
correlation between the protons of H-2 and H-3 indicated a eis relationship of the
B ring and the hydroxyl group. This was confirmed due to the lack of a 7 Hz vicinal
H-H coupling constant, which would be consistent with a trans relationship
(Palazzo de Mello et al., 1996). The *H and 13C NMR data showed proton and
carbon shifts, which are in accordance with literature values (Delle Monache et al.
1992) (Table 11.1).
Table 11.1. 1H, 13C NMR data and long-range correlations (HMBC) of
ourateacatechin (1) in MeOD
No. ,3cr, 1H. 5 ppm, HMBC
8 ppm (Jm Hz) correlations
2 79 46 d 4.77 br s 3.4, 9, 1s, 2-
3 67.24 d 4.19 br s 10
4 a 29 03 t 2.74 dd (16.8, 2.6) 2,3, 5,9, 10
ß 2.87 dd (16.8, 4.4)5 157.77 s - -
6 96.40 d 5.97 d (2.2) 5, 8, 10
7 157.42 S - -
8 95.86 d 5.93 d (2.2) 6, 7. 10
9 157.00 s - -
10 100.03 s - -
r 136.45 s - -
276' 107.12 d 6.55 s 2, 3', 4', 27=6'
3',5' 151.15 s - -
4' 135.94 s - -
OMe 60.77 q 3.79 s 4!
*
Multiplicities determined by DEPT sequences
Physical data of ourateacatechin (1). Brown-orange amorphous powder: [a]D -
47.3 ° (MeOH, c 1.00, 24 CC); UV Anax (MeOH): 271, 209 nm; positive EI-MS (70
eV) m/z : 322 [M+2H]+, 183, 168, 154, 140,33.
214
OCH,
2/6'
K„_7
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/A.
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Figure
11.11.13CINADEQUATEspectrum
ofourateacatechin
(1)
inMeOD/pyridine-d5(125MHz)
Phytochemistry
11.3 Triterpenes
11.3.1 Pristimerin
(20a)-3-Hydroxy-2-oxo-24-nor-friedelane-1(10),3,5,7-tetraen-carbonacid-(29)-
methylester (Bruning and Wagner, 1978)
= 464
C30H40O4
Compound 7 was found to have the molecular formula C3ûH40O4 by a combination
of mass spectrometry and 13C NMR experiments including DEPT, The 1H NMR
showed three olefinic protons (<5 6.35, 6.55 7.03) and seven methyl groups,
among them one vinyl methyl (5 2.21, C-23) and a methoxy group (5 3.66, C-31)
(Figure 11.12). Based on these data and the isolation of a remarkable quantity of
the compound (ca. 20% of the CH2CI2 extract) with an intensive orange color
suggested the presence of pristimerin (7). Pristimerin is a well-known pigment in
the root bark of plants of the Celastraceae (Grant and Johnson, 1957;
Habermeier, 1980). The comparison with the 1D NMR data with those of the
literature (Gunatilaka et al., 1989) confirmed the isolation of pristimerin (7), a
quinone-methide triterpene. The 1H and 13C chemical shifts are listed in Table
11.2 and 11.3, respectively.
219
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220
Phytochemistry
Physical data of pristimerin (7). Orange amorphous solid: [a]D - 179.0 °
(CHCI3, c
4.67, 24 °C); UV Amax (MeOH): 213, 423 nm; positive EI-MS (70 eV) m/z : 464, 267,
253,241,201,27.
11.3.2 Friedelane-3-on-29-ol
D:A-friedooleanan-29-ol-3-one (Itokawa et al., 1991), 29-hydroxyfriedelane-3-one
(Betancor et al., 1980), 3-oxofriedelane-29-ol (Patra and Chaudhuri, 1987)
31
Mr = 442
C30H50U2
8
Compound 8 was isolated as a white amorphous powder. The molecular formula
was obtained by EI-MS (M+ at m/z 442) as well as 13C NMR and formulated as
C30H50O2. The 1H NMR showed seven methyl groups attached to quaternary
carbons (Table 10.2 and 10.3). The partial spin systems obtained by COSY
spectra (H-1/H-2, H-1/H-10, H-4/H-23, H-6/H-7, H-18/H-19) were connected using
the aid of long-range correlations* and led to the identification of the known
friedelane triterpene 8 (Figure 11.14).
*Long-range correlations of HMBC measurements: H-4 to C-1/2/5, H-23 to C-
3/4/5, H-24 to C-4/5/6/10, H-25 to C-8/9/10/11, H-26 to C-8/13/14/15, H-27 to C-
12/13/14/18, H-28 to C-16/17/18/22, H-29 to C-19/20/21/30, H-30 to C-
19/20/21/29.
221
Phytochemistry
The stereochemistry was identified by ROESY experiments. The following
relevant correlations were observed: H-4/H-10, H3-23/H3-24, H3-24/H-8, H3-25/H-
8, H3-25/H3-26, H3-26/H-18, H,-26/Hr28, H„-27/H,-29, (Figure 1113)
Figure 11.13. ROESY correlations of compound 8 (300 MHz, CDCI3)
Physical data of fnedelane-3-on-29-ol (8). White amorphous solid [a]D ~ 25.2 °
(CHCI3, c 1.15, 24 °C); UV A^dX (MeOH) 212 nm, positive EI-MS (70 eV) m/z ; 442
[M*], 301, 273, 258, 246, 231, 216, 205, 188, 177, 163, 149, 83, 48
222
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Phytochemistry
11.3.3 2,3,7-Trihydroxy-6-oxo-1,3,5(10),7-tetraene-24-nor-
friedelane-29-oîc acid methylester
= 496
C3cH40O6
The main problem in the structure elucidation of the new compound 9 was the
identification of the weak signal C-7 of the quaternary carbon at 147.4 ppm in the
13C NMR. In addition, it did not show any long-range correlation in the HMBC
experiment due to the absence of the protons at C-5, C-9 and C-14 (Figure 11.
16). The 1H and 13C NMR data showed similar chemical shifts as those of the C, D
and E-rings of pristimerin (7) and were assigned with the aid of the HMBC and
COSY experiments (Figure 11.17; Tables 11.2 and 11.3). The stereochemistry
was identified by ROESY measurements (Figure 11.15). Comparing the 13C NMR
and 1H NMR data with a similar compound regeol C, the structure of compound 9
could be confirmed as 2,3,7-trihydroxy-6-oxo-1,3,5(10),7-tetraene-24-nor-
friedelane-29-oic acid methylester (Takaishi et al. 1997).
224
ppm
-110
-120
1-130
M40
-150
-160
M70
-180
5/1
4/1
3/1
62/1
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8/25$
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29/19
29/21
710
;
6:0
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Figure11.16.
PartofHMBC
spectrum
ofcompound9
inMeOD
(H/C:300/75.5MHz)
..-—
2.0
-r-^
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CL
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227
Phytochemistry
11.3.4 Celastrol
(20a)-3-Hydroxy-2-oxo-24-nor-friedelane-1(10),3,5,7-tetraen~carbonacid
= 450
^29''38^4
The comparison of 1H and 13C NMR data with those of pristimerin 7 showed a
close similarity between these two molecules (Table 11.2 and 11.3). The only
difference the methoxy group (5 3.55, C-31), which not present in compound 10.
The mass spectra exhibited the molecular peak at m/z 450 and confirmed the
molecular formula C29H3804 as well as the absence of the methyl group. The
relative stereochemistry was determined by a ROESY experiment. The
correlations between H-18/H3-28, H3-25/H3-26 and H3-26/H-18 indicated diaxial
relationships of these protons. Therefore ring fusions C/D and D/E were trans and
eis, respectively. Compound 10 was identified as celastrol, a pigment common to
many genera of the Celastraceae (Gisvold, 1939; Kutney et al. 1981: Morota et
al., 1995). In comparison with pristimerin (7) (6 g), Celastrol (10) was isolated in
very small quantity (2.1 mg).
Physical data of celastrol (10): Orange-yellow amporphous powder: [a]D - 23.0 °
(MeOH, c 1.00, 24 °C); UV Amax (MeOH): 213, 423 nm; positive EI-MS (70 eV) m/z.
450,436,253,241,202, 149, 83.
228
Phytochemistry
Table 11.2.1H NMR data of compounds 7, 8,10 in CDCI3 9 in MeOD (J in Hz)
H 7 (300 MHz) 8 (300 MHz) 9 (300 MHz) 10 (500 MHz)S ppm
1 6.55 s 1.97 m, 1.69 6.86 s 6.50 s
2 - 2.38 dd (3.3, 1.9)2.42 dd (3.2, 1.9)
- -
4 - 2.25 m - -
6 7.03 d (6.9) 1.777
1.31
- 7.06 d (7.0)
7 6.35 d (7.1) 1.50\1.42'
~ 6.33 d (7.2)
8 - 1.40} - -
10 - 1.53' - -
11 1.86°, 1.477 2.16°, 2.14 m,
2.17° 1.34J 1.97 dt (4.2, 14.2) 1.81e
12 1,81°, 1.35' 1.82 m, 1.87:,1.68* 1.66 dt (4.1, 14.1) 1.65"
15 1.65°, 1.56\ 2.88 m, 1.59",1.57° 1.32° 1.85° 1.54°
16 1.52°, 1.60°, 1.91', 1.81*.
1.89° 1.34-' 1.45' 1.47*
18 1.59° 1.62' 1.59 d (7.8) 1.58'
19 2.42 d (15.6) 1.50', 2.47 brd (15.5), 2.50 d (2.5),1.223 1.71 dd (7.4, 14.7) 1.75'
21 2.20°, 1.37' 2.17,
2.25',
1.37° 1.43' 1.36'
22 2.05°, 1.395, 2.12 dt (4.1, 13.8), 0.94 m.
0.97° 1.00' 0.96 m 2.10m
23 2.21 s 0.89 d (6.3) 2.57 s 2.21 s
24 - 0.73 s - -
25 1.45 s 0.88 s 1.49 s 1.44 S
26 1.26 s 1.04 s 1.37 s 1.26 s
27 0.53 s 1.05 s 0.69 s 0.59 s
28 1.09 s 1.22 s 1.08 s 1.10 s
29 - 3.29 d (10.7)3.25 d (10.7)
- -
30 1.17 s 1.03 s 1.15s 1.28 s
OCH, 3.55 s - 3.55 s
1H chemical shifts were assigned on the basis of HSQC experiments
0
Signal multiplicity was not described due to signal overlap.
229
Phytochemistry
1 able 113 13C NMR data of compounds 7, 8 10 in CDCI3 and
9 in MeOD (75 5 MHz, <Sppm)
Ca 7 8 9 10
1 1195 d 22 3 t 109 0 d 120 5 d
2 178 2 s 41 5 t 151 7 s 178 3 s
3 146 0 s 213 2 s 143 5 s 146 9 s
4 1170 s 58 2 d 127 9 s 1183 s
5 127 3 s 42 1 s 119 9s 127 5 s
6 133 9 d 41 3 t 182 8 S 135 2 d
7 1180 d 182 t 147 4 s 120 1 d
8 169 9 s 53 4 d 139 1 S 172 4 s
9 42 8 s 37 4 s 40 7 s 43 0 s
10 164 6 s 59 4 d 152 8 s 165 0 s
11 33 5 t 35 6 t 34 2 t 33 7 t
12 29 6 t 30 6 t 30 5 t 29 3 t
13 39 3 s 39 9 s 40 0 S 39 3 s
14 44 9 s 38 2 s 47 2 s 45 3 s
15 28 5 t 32 7 t 29 6 t 28 7 t
16 36 3 t 35 8 t 37 8 t 36 3 t
17 30 4 s 30 5 s 31 1 s 30 6 s
18 44 2 d 41 8 d 45 4 d 44 3 d
19 30 8 t 29 7 t 31 9 t 31 Ot
20 40 3 s 33 1 s 41 7 s 39 9 s
21 29 8 t 27 8 t 30 9 t 29 5 t
22 34 7 t 39 5 t 36 2 t 34 5 t
23 101 q 68q 140 q 104 q
24 - 146q . -
25 38.2 q 17.9 q 41 6 q 38 3 q
26 21 5 q 20 7 q 196 q 21 5 q
27 182 q 184 q 20 6 q 187 q
28 31 5 q 32 1 q 32 1 q 31 5 q
29 178 6 s 74 8 t 180 7 s 182 2 s
30 32 6 q 25 8 q 32 9 q 32 4 q
OCH, 51 4 q — 52 2 q —
1 Multiplicities determined by DEPT sequences
230
Phytochemistry
11.4 3,15-Dihydroxy-18-norabieta-3,8,11,13-tetraene
M =302
C19H2603
11
The structure elucidation is described in publication IV. 1H NMR and 13C NMR are
shown in Figure 11.18, The stereochemistry was elucidated by a ROESY
experiment. Due to the lack of correlation between H-5 and the protons of C-20,
A/B trans configuration was indicated and confirmed by comparison with literature
data (Nakano etal., 1997). Compound 11 showed an enol-form on position C-
3/4, but no keto function was detected in the 13C NMR spectrum (Kalinowski et al.,
1984). However, the "enolising" of the carbonyl group is generally a slow
reaction, the isolated compound 11 was stabile. The new triterpene 9 also
showed an enol-form on position C-7/8. In this case, the enol-form was favored
due to an expanded conjugated system (keto group on C-6).
231
oCM
h-
CD
CO
Q.
CL
| CM
JQ )CM
*>-CO ~^>
"7 O
\ Ie0
J I1-» »
-____ „___
___^
L1 LO
Lo
"3-L
1 LO
"xf
co
^IcLw CD
Ç\J ""^~"^=3~
CM-^
EQ.
CL
o"CM
232
Phytochemistry
12 Biological activities of isolated compounds
12.1 Cytotoxicity
Cardiac glycosides are known to be cytotoxic. The isolated cardenolides (2-6)
showed KB-cytotoxicity in a range of 0.074-0.199 iimol/ml. Our structure-activity
comparison indicated that the difference of one sugar in the chain resulted in a
significant difference in KB-cell cytotoxicity. Furthermore, the introduction of a C-
5ß hydroxyl group increased the cytotoxic activity(publication IV). The
cardenolides isolated from Euonymus alata (Thunb.) Sieb, exhibited KB-cell
activity in a range of 0.186-1.812 umol/ml. The genins of these compounds
showed a methyl group on C-19, a proton on C-5 and a hydroxyl group on C-1
instead of C-11. By a comparison of these cardenolides with different numbers of
sugars moieties, the cytotoxic activities decreased when the numbers of sugars
increase (Kitanaka et al., 1996).
Cassady and Suffness (1980) discussed the anti-tumor activities of the
cardenolides and structure-activity relationships in the KB cell cytotoxicity system
screened at the National Cancer Institute (NCI). The conclusions about structural
requirements for cytotoxicity in the KB system are the following:
There is some influence of the C-3 substituent on cytotoxicity with glycosides
being slightly more active than aglycones.
There is no substantial difference between a hydrogen and hydroxyl group at
C-5ß with regard to cytotoxicity (contradictory to our results, see above).
Those compounds with a hydrogens at C-5 retain activity but are notably less
cytotoxic than those with ß hydrogens at C-5.
Compounds with C-19 hydroxyl groups retain activity but are somewhat less
active than the C-19 aldehydes, while a C-19 acid is inactive.
233
Phytochemistry
* Substituents, which are tolerated without large changes in cytotoxicity, are 1ß-
OH, 11a-OH, and 12ß-OH groups.
4 C-14 requires a ß hydroxy group for maximum cytotoxicity.
Cardenolides are lacking therapeutically relevant cytotoxic activity in in vivo
systems. The activity is not easily reproducible and in the range of the toxic dose.
Consequently, these compound can not be used therapeutically. From the
volume of negative or extremely marginal in vivo data on cardenolides and the
large number of compounds tested at the NCI, it is apparent that there is little if
any prospect of developing therapeutically useful anti-cancer compounds from
this group (Cassady and Suffness, 1980).
Pristimerin (7) and celastrol (10) showed cytotoxicity in KB ceil (IC50 = 0.28, 0.72
fig/ml). According to Schwenk (1962) cytotoxicity may be dependent on the
quinoid structures. The reduction of the quinoid substances in the cells to
hydroquinones, which by autoxidation give hydrogen peroxide, may destroy
tumor cells by inhibition of glycolysis. Campanelli et al. (1980) studied the binding
of tingenone (quinone methine) to DNA. They suggest a possible mode of cellular
interaction, which may elicit anti-tumor activity. These two propositions show that
mechanism of the cytotoxic potential of quinone methides is still unclear.
Cytotoxicity of pristimerin (7) and celastrol (10) was also noted in HeLa cells
used for the NF-KB-test. Forty minutes after adding pristimerin (7), the cells
changed their morphology. After adding celastrol (10) the same phenomenon
was observed after 60 minutes. Gonzales et al. (1987) reported the ID50 value of
pristimerin (7) in HeLa cells as 0.6 ^g/rnl, and therefore it is similar to the value of
the cytotoxicity against KB cells.
Friedelane-3-on-29-ol (8) was not cytotoxic in KB cell (IC50 > 20 (ig/ml), but similar
compounds were reported to have cytotoxicity: 3-oxo-friedelane-29-oic acid (A-
234
Phytochemistry
549 lung carcinoma cells: ED50 = 3.7 uq/ml) and 3-oxo-friedelane-28-oic acid (KB
cells: ED50 = 3.7 ug/ml; L-1210: EDS0 = 2.95 ug/ml) (Mahato and Sen, 1997).
12.2 Other activities
Antibacterial and antifungal activity: The isolated compounds were tested against
several gram-positive and gram-negative bacteria, namely Bacillus cereus,
Staphylococcus epidermidls, Micrococcus luteus, Pseudomonas aeruginosa and
Escherichia coli. Candida albicans was chosen as a model for testing the
compounds against fungi. The results are summarized in publication IV. No
activity was detected agains gram-negative bacteria and against C. albicans.
NF-kB activity. At a concentration of 0.125 jig none of the isolated compounds
were active in the NF-kB assay. As previously mentioned, pristimerin (7) and
celastrol (10) showed cytotoxic activity in HeLa cells. The cytotoxic effect of
pristimerin (7) was probably responsible for the "positive" NF-kB test of the non-
polar extracts seen in the EMSA shift experiments (chapter 10.1).
Other activity. Pristimerin (7) was further tested against the following gram-
negative strains or parasites (methods: publication III).
Microorganism Result
Helicobacter pylori
Campylobacter jejuni
Plasmodium falciparum K1
Plasmodium falciparum T9/96
Trypanosoma brucei
Trypanosoma cruzi (epimastigotes)
Giardia duodenalis
1 mm, inhibition zone (600 ug tested)
not active (600 ug tested)
IC50 = 0.41 ug/ml
IC50 = < 0.41 |jg/ml
MIC = 0.12 ug/ml
MIC = < 3 jig/ml
MIC = 6.3 ug/ml; IC50 = 2.1 ,ug
Pristimerin (7) showed good anti-parasitic activity in all anti-parasitic assays. This
phenomenon is probably due to the unspecific and high KB cell cytotoxicity. The
activity against H. pylori and C. jejuni is very weak.
235
Phytochemistry
Ourateacatechin (1) was tested against H. pylori, C. jejuni, G. duodenalis and P.
falciparum but no activity was found. In the protein kinase screening, the catechin
did not show any inhibition of the kinases tested (chapter 6.3).
Friedelane-3-on-29-ol (8) showed no activity against G. duodenalis.
The cardiac glycoside (2-6) did not show any activity against P. falciparum.
13 Biomedicine, a way to explain the popular use of C, gaumeri ?
13.1 Gastrointestinal problems
Gastrointestinal problems are one of the major health problems among Yucatec
Maya. Especially children are affected by diarrhea (mal de ojo), flatulence and
worms. Adults are frequently medicated for diarrhea, dysentery, vomiting,
flatulence, cramps, indigestion, stomachache and tip'te' (publication I). The most
frequent pathogenic organisms, which cause gastrointestinal problems, are
bacteria, viruses and parasites. Some of the microorganisms causing
gastrointestinal problems are listed in Table 1 of publication III. In addition, Vibrio
cholerae induces cholera, which often leads to death due to dehydration.
Shigella dysenteriae and Entamoeba histolytica causes dysentery and Ascaris
lumbricoides (eelworm) are responsible for flatulence, diarrhea, lost of weight as
well as obstipation.
The antibacterial activity of pristimerin (7) seems to be an explanation of the use
of C. gaumeri as an anti-diarrheal medicine. Also the growth inhibition of G.
duodenalis by pristimerin (7) is of relevance for understanding the indigenous
use in treating diarrhea, pain, and indigestion. The spasmolytic effect of
ourateacatechin (1) supports the use of this species as a remedy for
gastrointesinal problems.
236
Phytochemistry
13.2 Snake bites
Venomous snakes are common in the
lowland Maya area. Three snakes of the
family of the Crotalidae, Crotalus
durissus (rattlesnake, tzabcan), Bothrops
asper (Fer-de-lance, taxinchan) and
Agkistrodon bilineatus {uolpoch) are the
most feared and respected snakes
among the Maya today. Their poisons
contain multi-protein complexes,
enzymes that support the distribution of
the venom, as well as digestive
enzymes. The toxins of the snakes can
be classified in:
a-toxins (postsynaptic inhibitory activity): Peptides with S-S bridges
%-toxins (neuronal inhibitory activity)
* ß-toxins (presynaptic inhibitory activity): Heterogeneous proteins and oligo¬
peptides, similar to phopholipases
* toxins which support presynaptic activity: Built of 57-60 amino acids
Na+-chanal-activator: Built of 39-43 amino acids
Hemorrhagines (metallo-proteases): Mr 20,000-100,000
For the three species of snakes, discussed here the effects of ß-toxins, Na+ -
chanal-activator and hemorrhagines are important. The ß-toxins destroy the
structure of the membranes and change the permeability of ions. As a result,
phospholipase A2 is able to pass the membranes. Depolarisation leads to the
contraction of the heart, skeletal and smooth muscle cells. In addition, some of the
ß-toxins are able to destroy the membranes of muscle cells. As a consequence,
Figure 13.1. Maya hunterbitten
by a rattlesnake (Lee, 1996),
237
Phytochemistry
myoglobinuria occurs. Hemorrhagines (metallo-proteases) destroy the
membranes of the capillaries and/or the inhibitors of the blood plasma.
Consequently blood extravasates to the tissues. Thus, the symptoms are
ecchymosis at the bite, paresthesia around the mouth, twitches, weakness,
syncope, sweating and vomiting. In case of death, shock, and collapse of the
cardiovascular system are observed (Teuscher and Lindequist, 1994).
13.2.1 C, gaumeri used in the treatment for snake bites
A two-cm piece of fresh root is chewed after a venomous snake bite. A piece of
this length weights about 0.8-1.6 g (o = 0.5-1 cm). Thus, 8-16 mg (1 % of dry
weight) of pristimerin (7) 0.16-0.32 mg (0.02 %) of ourateacatechin (1) and 0.08-
0.16 mg (0.01 %) of cardiac glycosides (2-6) are consumed. The other isolated
compounds (8, 9, and 10) are not discussed further because they occur in little
amounts and no strong and relevant activity is expectable.
Also the powdered root mixed with water is used topically. The healers mentioned
that the remedy takes away the pain, reduces the inflammation and "adsorbs
venom". From a biomedical perspective no clear answers can be found, why C.
gaumeri is traditionally used to treat bites. To get an answer, further
pharmacological studies with the plant and their main principles must be carried
out. However some hints about active compounds against snake bites are found
in literature (Houghton and Osibogun, 1993; Mors, 1991).
Persimmon tannin from Diospyros kaki L. f. (Ebenaceae) neutralized
neurotoxic and haemorrhagic venoms. The results were very promising.
Consequently the authors recommended the extract as a washing agent in
the emergency treatment of snake bite wounds. The structure of the D. kaki
tannin was reported as being made up of catechin and gallocatechin units.
Protocatechuic acid from Cryptolepis sinensis Merr. (Asclepiadaceae)
was isolated. It deactivated venom and
238
Phytochemistry
gymnemic acids of the leaves of Gymnema sylvestre R. Br. (Asclepiadaceae)
inhibited ATPase.
* The roots of Aristolochia spp. (Aristolochiaceae) rich in aristolochic acid
deactivate various snake venoms.
The leaves of Strophanthus hispidus (Apocynaceae) prolonged the time
taken to clot for blood treated with the venom of Echis cahnatus, which
causes intra-arterial clotting of blood.
# Cabeca-de-negro (negro's head) is supposed to be the main ingredient of the
oral anti-snake-venom remedy Especffico Pessoa, which is manufactured in
north-east Brazil. Two prenylated pterocarpans were isolated from the
remedy and shown to be active against the toxic cardiovascular effects of
Bothrops venom (Hostettmann et al., 1997).
Features which could possibly be of importance in the treatment of snake bites
are the acidic nature and/or catechol grouping, Polyphenolic substances are well
known to bind on proteins and to inhibit enzymes. Ourateacatechin (1), a
polyphenolic substance, presumably has the ability to inhibit the toxic enzymes of
snake venom by topical application. Generally, catechines show anti-edematic
activities and act as diuretics (Hansel et al., 1999). In case of ourateacatechin,
this effect could help to treat the local and expanded tumescence after a bite of C.
durissus and B. asper. Furthermore, the treatment of the edema could help to
avoid or to diminish necrosis and ecchymos. The mechanisms of the catechins
are assigned to the anti-oxidative property and the inhibition of enzymes (Hansel
etal., 1999).
When the plant remedy is used in the form of a plaster it probably prevents
secondary infections of the bite. This could well be due to pristimerin (7), which
showed high antibiotic activity.
Anti-inflammatory activity is a property common to many plants claimed to be
active against snake bites. But at the test concentration used no compound
isolated from C. gaumeri showed any activity in the NF-kB assay.
239
Phytochemistry
13,2.2 What have the cardenolides to do with snake bites?
Cardiotonic steroids and their activity: Cardenolides are known for their positive
inotropic effect on cardiac muscle by increasing contractility and decreasing the
frequency of heart beat via Na7K+-ATPase inhibition and increasing delivery of
Ca2" to cardiac muscle cells. Cardiotonic steroids have a very narrow therapeutic
window. The emetic dose of cardenolides is approximately 50 % of the lethal
dose. Fatal poisoning after oral ingestion of plants is rare due to the bitter taste
and the subsequent vomiting. The resorption rate of ouabain (g-strophanthin) by
peroral application is less than 5 % due to the substance's polarity (five hydroxyl
groups). Digitoxin with one hydroxyl group is resorbed practically completely (100
%). The isolated compounds (2-6) have two to three hydroxyl groups and it can
be supported that their resorption rate most likely lie between those of ouabain
and digitoxin. The resorption rate furthermore depends on the sugar moieties,
which can be optimized by increasing the lipophilic character (separation of
sugars, etherification and esterification of hydroyl groups). The study on the
structure-activity relationships (SAR) of the cardenolides on the Na7K+-ATPase is
summarized in the following list and show some interesting correlations to the
SAR of cytotoxicity (Malcolm, 1991; Thomas 1992) (see chapter 12.1).
A lactone function (C-17) and sugar side chains (attached to C~3) are not
necessary, but contribute considerably to the selectivity and activity of
cardenolides. The compounds containing 6-deoxy sugars are the most potent
of all cardiotonic steroids. The addition of an extra sugar unit to a monoside
leads to a decrease in activity.
14ß is a prerequisite for receptor recognition and high potency (C/D eis
junction). Trans configuration prevents recognition by Na7Kr-ATPase.
* 5a (A/B trans) is more active than 5ß (A/B eis)
Polarity (e.g. OH-groups on C-1, C-5, C-11, C-12) influences the absorption
from the intestine, but not necessarily the activity.
240
Phytochemistry
Cardenolides in the therapy of snake bites: As mentioned before, a two-cm piece
of root of C. gaumeri contains approximately 0.08-0.16 mg cardiac glycosides. A
standard initial dosage for treating heart insufficiency with digitoxin lies between
1.0-1.5 mg, afterwards a dosage of 0.25 mg every 6 h is prescribed. Thus, the
herbal medicine reaches the therapeutic dosage.
Cardiac insufficiency and cardiac arrhythmia caused by snake venom are not
described in Mexico and Middle America. However failure of cardiovascular
system occurs due to hypovolemic and hemorrhagic shock (Junghanss and
Bodio, 1996). The cardenolides have a positive influence on the cardiovascular
system. The stimulation of the Nervus vagus by the cardiac glycosides decreases
the frequency of the heart and the velocity of the atrioventricular conduction (AV).
This effect is due to an increased KT permeability and thus stabilizes the
membrane potential. As it is well known, cardenolides also exert a positive
inotropic effect on the heart muscle. In consequence, they improve the circulation
of a patient with an insufficient heart and they probably can help to avoid
hypovolemic shock. The plant remedy and the cardenolides are certainly not a
specific antidote against snake venom, but influence the cardiovascular system in
a positive way. Caffeine or coffee is given to patients bitten by a venomous snake
to stimulate the central nervous system (CNS) and in particular, the respiration
(Falbe and Regitz, 1992). Could it be that the cardiac glycosides also have a
general stimulatory effect on the CNS? The mechanism may be a similar one as
observed with local anesthetics or alcohol. These latter agents stabilize the cell
membranes in general and restrain the inhibitory pathways in the CNS resulting
in a predominance of the excitatory neurons. Should the cardenolides have a
similar general stabilizing effect on the membranes in the CNS, one can
hypothesize that the inhibitory pathway may be affected first and this would also
lead to a predominance of the excitatory systems, including stimulation of
respiration. It is well known that the cardiac glycosides accumulate strongly in the
CNS. Indeed, cardenolides often induce side effects in the CNS such as general
241
Phytochemistry
excitement, xanthochromia and hallucinosis in heart patients. In order to further
test this hypothesis in vivo tests should be carried out.
It is of interest that the Yucatec Maya use two other important plant species for the
treatment of snake bites. These are Urechites andrieuxii and Echites
yucatanensis, both belonging to the Apocynaceae. The Apocynaceae are known
to contain cardenolides and therefore strengthen the hypothesis that these
compounds could potentially be useful in the treatment of snake bites.
242
Publication IV
Cytotoxic cardenolides and antibacterial terpenoids from
Crossopetalum gaumeri
Anita Ankli3, Jörg Heilmann3, Michael Heinrich5 and Otto Sticher1
a
Department of Applied BioSciences, Institute of Pharmaceutical Sciences, Swiss
Federal Institute of Technology (ETH) Zurich, Winterthurerstr. 190, CH-8057
Zürich, Switzerland
b Centre for Pharmacognosy and Phytotherapy, The School of Pharmacy, 29/39
Brunswick Sq., London WC1N 1 AX, UK
Accepted in
Phytochemistry 2000
Publication IV
Abstract
From the methanol extract of the roots of Crossopetalum gaumeri four new highly
cytotoxic cardenolides, securigenin-3ß-0-ß-6-deoxyguloside (2), 19-hydroxy-
sarmentogenin-3ß-0-ß-6-deoxyguloside (4), sarmentogenin-3ß-0-[a-allosyl-(1 ->4)-
ß-6-deoxyalloside] (5), securigenin-3ß-0-[a-allosyl-(1->4)-ß-6-deoxyalloside] (6)
were isolated. The dichloromethane extract afforded the new diterpene 3,15-
dihydroxy-18-norabieta-3,8,11,13-tetraene (7) as well as the new triterpene 2,3,7-
trihydroxy-6-oxo-1,3,5(10),7-tetraene-24-nor-friedeIane-29-oic acid methylester
(11). The new terpenoids lack cytotoxcity and the antibacterial activity is moderate
to low.
Keywords
Crossopetalum gaumeri; Celastraceae; cardenolides; triterpenes; diterpene;
cytotoxic activity; antibacterial activity; Yucatec Maya; traditional medicine.
244
Publication IV
Introduction
Based on an ethnobotanical field study with the Yucatec Maya and an evaluation of
their medicinal plants, the roots of Crossopetalum gaumeri (Loes.) Lundell were
chosen for a detailed phytochemical study (Ankli et al., 1999; Ankli et al.,
submitted). A piece of root is chewed after a person has been bitten by a snake,
the pulverized root is mixed with water and is put on the wound in form of a plaster.
Furthermore, the decoction is used orally for diarrhea. Crossopetalum is a genus in
the family Celastraceae with 36 species in tropical America (Mabberly, 1987).
Especially due to the discovery of the antitumor effect of maytansinoides and other
novel structure types isolated from Celastraceae, this family is of great interest for
phytochemical investigation (Bruning & Wagner, 1978).
However, the genus Crossopetalum is not well investigated. From C. tonduzii
sesquiterpenes (Tincusi et al., 1998) and from C. uragoga used by the Huastec
Maya as an anti-diarrheal medicine triterpenes have been isolated (Dommguez et
al., 1984). This report describes the isolation, structure elucidation, as well as
cytotoxic and antibacterial activity of five cardiac glycosides (2-6), four terpenoids
(7-11), and one catechin derivative (1) from the methanol and dichloromethane
extracts of the roots of C. gaumeri.
Results and discussion
The methanol extract was fractionated using cytotoxicity against a KB cell line as a
lead. This led to the isolation of the well known ourateacatechin (1) (Drewes &
Mashimbye, 1993) and five cardenolide glycosides (2-6). All cardenolides were
isolated as white amorphous powder and showed greenish to blue spots on TLC
after spraying with vanillin-H2S04 as well as an intense fluorescence under UV 366
nm.
245
Publication IV
2 ß-6-deoxygulosyl H CHO
3 ß-6-deoxygulosyl OH CHO
4 ß-6-deoxygulosyl H CH2OH5 a-allosyl-(1-^4)-ß-6-deoxyallosyl H CH36 <x-allosyl-(1->4)-ß-6-deoxyallosyl H CHO
The 1H NMR spectrum of compound 2 showed characteristic signals of a
butenolactone ring at 8 5.03, 4.92 (each dd, J ~ 18.3, 1.5 Hz, H-21a and b) and
5.92 (s, H-22), as well as a singlet proton at 5 9.41 indicating a cardenolide with an
aldehyde function. A doublet at <5 4.67 (J= 8.2 Hz, H-1'), four additional protons
between 3.99 - 3.46 ppm and a signal at S 1.21 (d, J = 6.7 Hz, H3-6') pointed to the
presence of a ß-linked deoxyhexose (see Table 1). 13C NMR experiments,
including DEPT 135, sorted 29 carbons into two methyl, nine methylene, 13
methine and five quaternary carbons which is consistent with the molecular formula
C29H42O10 (see Table 2). FAB-MS spectrum showed pseudomolecular peaks at m/z
551 [M+H]+and at m/z 573 [M+Na]+. Fragments at m/z 413 and 146 confirmed the
sugar to be a deoxyhexose (Ferth & Kopp, 1992; Habermeier, 1980). In
accordance with the proposed molecular formula positive HRESI-MS revealed a
pseudomoleculare ion at 551.2849 [M+H]+. Based on COSY, HSQC, HMBC and
ROESY experiments and in comparison to literature data the aglycon was
identified as securigenin (Kawaguchi et al., 1993; Ferth et al., 1992). Structure
elucidation of the sugar moiety was performed on the basis of coupling constant
analysis and of ROESY and COSY experiments. H-2' resonated at 3.60 ppm as
246
Publication IV
double doublet indicating an axial-axial relationship to H-1' (J = 8.2 Hz) and an
axial-equatorial to H-3' (J = 3.5 Hz). Due to the small coupling constants of H-4'
(dd, J = 3.5 and 1.0 Hz) and a strong NOE between H-1' and H-5', H-4' is in
equatorial and H-5' in axial position. Based on these data the sugar moiety was
identified as ß-6-deoxygulose, which was confirmed by TLC after hydrolysis of 2
and comparison to authentic ß-6-deoxygulose. Consequently, compound 2 was
identified as the new securigenin-3ß-0-ß-6-deoxyguloside (2). COSY, HMBC and
HSQC experiments showed unambiguously that C-2' resonates at <5 68.1 and C-4'
at 8 72.3 (Table 2). Therefore, the 13C chemical shift assignment for ß-6-
deoxygulose in literature must be corrected (see Table 2).
The 13C NMR spectrum of compound 3 showed close similarities to compound 2
with the exception of the signal of C-5 which resonated as quaternary carbon at c5
74.0 and downfield shifts of C-4 and C-6 pointing to an additional substitution with
a hydroxyl group. In accordance the positive FAB-MS spectrum exhibited a [M+H]+
peak at m/z 567 and a fragment at 421 [M+H-deoxyhexosyl]\ After extensive 1D
and 2D NMR analysis compound 3 was identified as sarmentosigenin-3ß-0-ß-6-
deoxyguloside. It was described for the first time as canescein from the genus
Erysimum (Maslennikova et al., 1967; Makarevich & Kovalev, 1968).
The molecular formula of compound 4 was established as C29H44O10, obtained from
the 13C NMR spectrum and the positive FAB-MS, showing a pseudomolecular peak
at m/z 553 [M+H]+. In contrast to securigenin-3ß-0-ß-6-deoxyguloside (2), in the 1H
NMR spectrum of 4 the aldehyde proton is replaced by two protons resonating at 8
3.86 and 3.71 (each d, J = 10.8) pointing to a hydroxymethyl group. As a result of
MS and NMR analysis, the genin was identified as 19-hydroxysarmentogenin
(Kopp & Kubelka, 1982). The sugar rest was identified as ß-6-deoxygulose as
described for compound 2, Thus, compound 4 is the hitherto unknown 19-hydroxy-
sarmentogenin-3ß-0-ß-6-deoxyguloside.
The 1H and 13C NMR of compound 5 indicated the presence of a cardenolide with
two sugar moieties and a C-19 methyl group (see Tables 1 and 2). The DEPT
experiments sorted 35 carbons into three methyl, ten methylene, 17 methine and
247
Publication IV
five quaternary carbons. The positive ESI-MS spectrum gave a molecular ion at
m/z 698. Therefore, the molecular formula of 5 was determined as C35H54014 and
the aglycon was identified as sarmentogenin (Hanada et al., 1992). The identity
and connectivities of the sugar moiety was deduced from a combination of 1D and
2D NMR experiments (1H,1H COSY, HSQC, HMBC and ROESY) and confirmed by
TLC hydrolysis and comparison with authentic compounds. Therefore, 5 was
established as the new sarmentogenin-3ß-0-[a-allosyl-(1->4)-ß-6-deoxyallosideJ.
The aglycon of compound 6 showed nearly the same chemical shifts and
correlations in the 1D and 2D NMR spectra as compound 2 and was therefore
identified as securigenin. The chemical shifts and connectivities of the sugar
moieties were identical with those of compound 5. Therefore, 6 was identified as
the new securigenin-3ß-0-[a-allosyl-(1-»4)-ß-6-deoxyalloside].
All isolated cardiac glycosides 2-6 showed high cytotoxic activity against a KB cell
line (see Table 3). It is worthy of note that the cytotoxicity is not correlated with the
oxidation status of the C-19 methyl group. Cytotoxicity of compounds 2 and 4 (IC50
0.164 umol 2 vs. 0.199 jimol 4) as well as 5 and 6 (IC50 0.075 jimol 5 vs. 0.104
umol 6) were in the same range. Comparison between compounds with identical
genin and different sugar chain revealed significant difference (IC500.164 umol 2
and IC50 0.104|imol 6). Introduction of a hydroxyl group at C-5 doubled cytotoxicity
(IC500.164 umol 2 vs. 0.074 umol 3).
248
Table
1
'HNMR
data
ofcompounds2-6(CD3OD,
8ppm,J
inHz,500MHz;compound
6,600MHz
)*
H2
34
56
12.
15*,
1.82*
2.47m,2.24m
2.25°,
1.84
*2.30brd(13
4),1.
50*
2.18
*,1.
82*
21.81
51.88*
1.82
*,1.
74*
1.82
*,1.
66*
1.80
*
3 4
4.06
brs
2.15°,
1.73°
4.21brs
2.09°,
1.73*
4.05brs
1.81*,
1.60
*
4.01brs
1.83
*,1.54°
4.06m
1.78
*,1.
70°
52.37m
-
2.11m
1.79°
2.36m
61.76°,
1.46°
1.88
*,1.
66*
1.78*,
1.27
°1.86
*,1.
27*
1.46*
71.82
°2.
07°,
1.29
*1.83*,
1.33
°1.
80*
1.82
*,1.
26*
81.82°
2.01*
1.72*
1.66*
1.84*
91.
82°
1.74
*1.
89*
1.80*
1.82*
r4.42m
3.95*
3.82
td(1
0.5
4.3)
3.72
td(1
0.4
4.3)
4.42m
12
1.75
°,1.50°
15
2.18e,
1.69°
1.68
*,1.50dd
(13.7,
10.7
)
2.16*,
1.69*
1.69m,
1.56brt(1
2.1)
2.25*,
1.78
°
1.68
*,1.
56*
2.21
*,1.75*
1.76
*,1.53brt
(11
2.18
°,1.70*
16
2.18°,
1.91°
2.15°,
1.90
*2.22*,
1.91
°2.19
*,1.
90*
2.17
°,1.
90*
17
2.931(7.1)
18
0.98s
2.931(7.1)
0.90s
2.93m
0.93s
2.91m
0.90s
2.93
t(6
.9)
0.98
s
2"
2.1 1 2
)9.41
s
5.03dd
(18.2
4.92dd
(18.2
Z5.92s
4.67d
(8.2
)3.60dd
(8.2
,
,1.5)
,1.5)
3.5)
9.97s
5.03dd
(18.4,
1.5)
4.92dd
(18.4,
1.5)
5.92s
4.70d
(8.1)
3.59dd
(8.2
,3.3)
3.86d
(10.8),
5.03dd
(18.
24.92dd
(18.
25.94d(1.0)
4.67d
(8.2)
3.62dd
(8.1,
v
3.71
d(10.8)
1.5)
1.5)
3.4)
1.07s
5.02d
(18.
3)4.91d(18.3)
5.91
s
4.67d
(7.9
)3.35°
9.42
s
5.02dd
(18.
5,1.5)
4.91dd
(18.
5,1.5)
5.90s
4.70d
(8.1
)3.37dd
(8.1
,3.1
)3 4 5
3.96°
3.46dd
(3.8
,3.99°
1.0)
3.96*
3.46d
(3.4)
4.01
q(7.2,
1.0)
3.98*
3.47d
(3.5)
3.99
*
4.33
t(2
.9)
3.27dd
(9.3,
3.85°
2.7)
4.341(
2.6)
3.28dd
(9.5,2.
6)3.84*
6 1 2
1.21d(6.7)
1.22
dd(6.6)
1.23d
(6.7)
1.28d
(6.1
)4.73d
(7.9
)3.
34°
1.29d(6.2)
4.74d
(7.7
)3.35°
3 4 5
,
4.05t(2.9)
3.54dd
(9.0,
3.69°
2.7)
4.05m
3.53dd
(8.8,2.
8)3.70°
6'
3.82
°,3.70*
3.82
°,3.70*
ro
10
"Ass
ignm
ents
areconfirmedbyCOSY,HSQCandHMBC,
*Multiplicities areuncleardue
tooverlapping
Table 2
13C NMR data of compounds 2-7,11 (CD3OD, 75.5 MHz, ö ppm)
Ca 2 3 4 5 6 Ca 7 1 1
1 23.5 t 19.7 1 27.5 t 33.8 t 24.9 t 1 34.6 t 109.0 d
2 21.5 t 25.5 t 28.0 t 28.3 t 26.9 t 2 26.0 t 151.7 s
3 72.8 d 73.6 d 74.4 d 75.9 d 74.4 d 3 163.1 S 143.5 s
4 29.1 t 34.0 t 31.4 t 31.6 t 30.4 t 4 119.0 s 127.9 s
5 30.6 d 74.0 s 32.0 d 39.1 d 32.0 d 5 46.4 d 119,9 s
6 28.8 t 37.1 t 27.7 t 28.1 t 30.1 t 6 21.1 t 182.8 s
7 25.5 1 23.9 t 22.2 t 22.7 t 22.9 t 7 24.6 t 147.4 S
8 41.4 d 40.6 d 41.7 d 41.9 d 42.8 d 8 124.1 s 139.1 s
9 41.2 d 44.5 d 42.7 d 43.0 d 42.6 d 9 148.3 s 40.7 s
10 52.1 S 55.1 S 41.1 s 37.5 s 53.5 S 10 36.8 s 152.8 S
11 66.3 d 66.9 d 69.0 d 68.9 d 67.7 d 11 115.7 d 34.2 t
12 48.8 t 48.5 t 50.6 t 50.5 t 50.2 t 12 123.5 d 30.5 t
13 49.6 s 49.5S 51.1 S 51.0 s 51.0 s 13 129.4 s 40.0 s
14 83.9 s 83.8 s 85.8 S 85.6 s 85.3 s 14 154.6 s 47.2 s
15 31.7 t 31.5 t 33.5 t 33.6 t 33.1 t 15 75.7 s 29.6 t
16 26.4 t 26.4 t 27.9 t 27.9 î 27.8 t 16 30.6 q 37.8 t
17 50.2 d 49.9 d 51.8 d 51.8 d 51.6 d 17 30.7 q 31.1 S
18 15.9 q 16.0 q 17.6 q 17.5 q 17.3 q 18 18.5 q 45.4 d
19 207.1 d 209.6 d 66.7 t 24.3 q 208.5 d 19 - 31.9 t
20 175.7 s 175.7 s 177.1 s 177.1 s 177.1 s 20 22.9 q 41.7 s
21 73.9 t 73.9 t 75.3 t 75.3 1 75.3 t 21 30.9 t
22 116.5 d 116.6 d 118.0 d 118.0 d 118.0 d 22 36.2 t
23 176.2 s 176.0 s 177.7 s 177.7 s 177.6 s 23 14.0 q
1' 99.1 d 98.5 d 100.0 d 99.9 d 99.8 d 24 -
2' 68.1 d 68.1 d 69.5 d 72.0 d 72.0 d 25 41.6 q
3' 72.2 d 72.1 d 73.5 d 72.4 d 72.4 d 26 19.6 q
4' 72.3 d 72.2 d 73.7 d 84.1 d 84.0 d 27 20.6 q
5' 68.6 d 68.8 d 69.9 d 69.4 d 69.5 d 28 32.1 q
6' 15.0 q 14.9 q 16.4 q 18.2 q 18.2 q 29 180.7 s
1" 103.5 d 103.5 d 30 32.9 q
2" 72.3 d 72.3 d 31 52.2 q
3" 73.1 d 73.1 d
4" 68.4 d 68.4 d
5" 75.2 d 75.2 d
6" 62.6 1 62.6 t
Multiplicities determined by DEPT sequences.
250
Publication IV
Table 3
Cytotoxicity of compounds 1-11 against a KB cell line (IC50 in umol)
Compound IC50 standard error n
1 5.938 ±0.1 4
2 0.164 ±0.015 4
3 0.074 ±0.009 4
4 0.199 ±0.008 6
5 0.075 ± 0.004 8
6 0.104 ± 0.005 6
7 > 66 - 4
8 >45 - 4
9 0.603 ±0.01 4
10 1.6 ±0.14 4
11 9.476 ±0.35 4
podophyllotoxin 0.014 - »4
Bioactivity-guided fractionation of the dichloromethane extract, using antibiotic
activity against Bacillus cereus, Staphylococcus epidermidls and Micrococcus
luteus led to the isolation of the new diterpene 3,15-dihydroxy~18-norabieta-
3,8,11,13-tetraene (7), the three known triterpenes friedelane-3-on-29-ol (8),
pristimerin (9) (Gunatilaka et al.. 1989), celastrol (10) (Kutney et al., 1981; Patra &
Chaudhuri, 1987), and the new triterpene 2,3,7-trihydroxy-6-oxo-1,3,5(10),7-
tetraene-24-nor-friedelane-29-oic acid methylester (11). Compound 7 was isolated
as a pale yellow amorphous powder. The 13C NMR spectrum showed signals of 19
carbons, which could be assigned with DEPT experiments into four methyl, four
methylene, three methine and eight quaternary carbons. The positive EI-MS,
showing a pseudomolecular peak at m/z 301 [M-H]~, in combination with the 13C
NMR spectra allowed the establishment of the molecular formula C19H2603. The
1H,1H TOCSY and COSY spectra revealed three spin systems. Spin system A with
two coupling protons at 8 6.79 and 6.91 (each d, J- 8.3, H-11 and H-12) indicated
the presence of a 1,2,3,4-tetra-substituted aromatic ring. Spin systems B (H-5/H-
6/H-7) and C (H-1/H-2) belong to a CH-CH2-CH2 and a CH2-CH2 moiety,
respectively. The linkage between the spin systems was established by a HMBC
experiment and led to a norabietan skeleton with a C-3/C-4 double bond (Takaishi
et al, 1997a). HSQC, HMBC and ROESY were utilized to clarify the 1H and 13C
251
Publication IV
NMR assignments and the stereochemistry. Taken together the data established
the structure 3,15-dihydroxy-18-norabieta-3,8,11,13-tetraene (7). 13C NMR data
clearly revealed that we isolated only the enol form 7 whereas the tautomeric
keton is completely absent.
7 11
The molecular formula of compound 11, a yellow amorphous powder, was
established as C30H40O6 obtained from the 13C NMR spectrum and from positive EI-
MS, showing the molecular peak at m/z 496 [M]+. The 13C NMR and the DEPT
revealed the presence of seven methyl, seven methylene, two methine and 14
quaternary carbons. The UV spectrum contained absorption maxima at 248 and
321 nm due to a phenolic ring system. The 1H and 13C NMR data showed similar
chemical shifts to those of the C, D and E-ring of pristimerin (9) (Gunatilaka et al.,
1989). For ring A and B, long range correlations of the proton H-1 with the carbons
C-2, C-3, C-4, C-5, C-6, C-10 and the protons H3-23 with C-1, C-2, C-3, C-4, C-5,
C-6 and C-10 were observed. These data and comparison with literature data of
regeol C (Takaishi et al., 1997b) led to the identification of 2,3,7-trihydroxy-6-oxo-
1,3,5(10),7-tetraene-24-nor-friedelane-29-oic acid methylester (11).
Confirming earlier published data the triterpenoids 9 and 1 0 showed high
antibacterial activity particularly against S. epidermidls as well as a remarkable
cytotoxicity against KB cells (Kutney et al. 1981; Gonzalez et al., 1998). Both new
252
Publication IV
terpenoids 7 and 11 lack cytotoxicity and the antibacterial activity is moderate to
low (see Tables 3 and 4).
The antibacterial activity of pristimerin (9) seems to be an explanation of the use of
C. gaumeri as an anti-diarrheal medicine. Due to its cytotoxic potential C. gaumeri
and preparations thereof should be used with great caution.
Table 4
Antibacterial activities of compounds 7-11 (MIC in umol)
Minimum inhibition concentration (MIC) in broth
Compound ——-
B. cereus S. epidermidls M. luteus
7 423.84
8
9 8.62 0.54 8.62
10 4.44 1.11 4.44
11 129.03 129.03 32.26
chloramphenicol 6.19 12.38 6.19
Experimental
General experimental procedures
Optical rotations were measured in MeOH or CHCI3 on a Perkin-Elmer model 241
Polarimeter. UV spectra were obtained on a Kontron-Uvikon 930
spectrophotometer, using MeOH as solvent. EI-MS were measured on a Hitachi-
Perkin Elmer RMUGM mass spectrometer at 70 eV. FAB-MS were obtained in the
positive mode on a ZAB 2-SEO spectrometer, using 3-nitrobenzylalcohol as matrix.
ESI-MS (positive mode) were measured on a TSQ 7000 mass spectrometer.
Applying HRESI-MS (positive mode) to the cardiac glycosides only compound 2
expressed a detectable [M+H]+ pseudomolecular peak. 1H and 13C NMR spectra
were recorded using Bruker AMX-300, DRX-500 and DRX-600 spectrometers. The
spectra were measured in CD3OD or CDCI3 and the residual CH3OH and CHCI3
resonances were used as internal references. For VLC, silica gel (60F254, 40-60
u,m, Merck) and RP-18 material (40-63 [im, CU Chemie Uetikon AG) was used.
253
Publication IV
MPLC was carried out using a Büchi chromatography pump B-688 and a 3.5 x 80
cm Büchi column packed with silica gel (60HF254, 15 j.im, Merck). HPLC
separations were performed with a Merck-Hitachi L-6200 intelligent pump
connected to a Merck-Hitachi L-4000 UV detector or a Waters model 590 pump
connected to a Pharmacia Biotech Uvicord Sil detector. HPLC columns were from
Knauer (Spherisorb S5 ODSII, 250 x 16 mm, EOT Chemie AG, 5 |im - for the
MeOH extract and Spherisorb ODSII, 250-20 mm. Waters, 10 |iin - for the CH2CI2
extract).
Plant material
C. gaumeri was collected in the villages and surroundings of Chikindzonot, Ekpedz
and Xcocmil, Yucatan, Mexico (1994-1995). Authenticated voucher specimens
were deposited at the Herbarium of the Centra de Investigacion Cientifica de
Yucatan (CICY) in Mérida, the National Herbarium of Mexico (MEXU), the Instituto
Nacional Indigenista (INI) in Valladolid, Yucatan, the ETH Zurich (ZT) and the
Centre for Pharmacognosy and Phytotherapy, The School of Pharmacy, London,
UK.
Extraction and isolation
Air-dried and powdered roots of C. gaumeri (2.26 kg) were successively extracted
with CH2CI2, MeOH, 70% aqueous MeOH and H20. Extraction with CH2CI2 yielded
42 g extract and from the methanol extraction 95 g were obtained. An aliquot of the
latter extract (42 g) was partitioned between CHCI3 and 60% aqueous MeOH (1:1).
The polar fraction was further partitioned between n-butanol and H20 (1:1). The
butanol fraction was combined with the CHCI3 one and applied to VLC, as two
separate portions, 10 g each. Elution with CHCI3 containing increasing amounts of
methanol yielded 18 fractions. Fraction 9 (CHCI3-MeOH, 9:1) yielded compound 1.
Fractions 13-15 (6.9 g, CHCI3-MeOH, 9:1 to 6:4) were combined and subjected to
an open column chromatography using silica gel 60 (35-70 |±m) as stationary
phase and increasing amounts of aqueous MeOH (MeOH-H20, 99:1) in CHCI3 as
254
Publication IV
eluent (100 % CHCI3 to 45 % CHCI3) to give 17 subfractions. The subfractions 2
(83.5:16.5), 5 (78:22), 7 (70:30), 11 (67:33) and 14 (50:50) were purified by HPLC
on RP-18 using ACN-H20 (20:80) as mobile phase for the subfractions 2, 5 and 7
and ACN~H20 (15:85) for the subfractions 11 and 14 yielding the compounds 2-6.
An aliquot (33 g, 35 %) of the CH 2CI2 extract was fractionated over silica gel
(VLC), using n-hexane-EtOAc mixtures with increasing polarity as eluent to afford
22 frs. Fraction 8 (80:20) contained compound 9. Frs 11 (30:70) and 12 (30:70)
were combined and subjected to normal-phase MPLC employing n-
hexane-EtOAc-MeOH (15:3:0.5 to 10:10:5) mixtures as mobile phase. Based on
TLC control the subfractions were combined to give 21 fractions. Fraction 3
(15:3:1) contained compound 9, after purification of fraction 6 (12:8:1) based on
MeOH-solubility, compound 8 was isolated. Fr 13 (10:10:3) was purified on a C18
Sep-Pak® Cartridge with 50% aqueous MeOH increasing the methanol proportion
to give 7. Fr 8 (10:10:1) was separated by RP-VLC using increasing amount of
MeOH in water as eluent to give 38 fractions. Of these, frs 24 and 25 (MeOH-H20,
9:1) were combined and refractionated by a silica VLC using n-hexane and n-
hexane-CHCI3 mixtures with increasing polarity which afforded 19 frs. Frs 10-15
(20:80) were combined and purified by RP-HPLC (ACN~H20, 9:1) to yield 10 and
11.
Detection of cardiac glycosides and hydrolysis of the cardiac glycosides on
TLC plates
As mobile phase for TLC analysis of the cardenolides EtOAc-MeOH-H20
(81:11:8) was used. For detection vanillin-H2S04 was sprayed on the TLC plates
(silica gel 60 F254) and heated at 110 °C for 5-10 min. The evaluation was carried
out under vis and under UV light 366 nm.
The TLC hydrolysis of the cardiac glycosides was realized following Kartnig &
Wegschaider (1971) with some modifications. The tank was not saturated with
36% HCl and the TLC plate was exposed to HCl vapor for 5 min (100 °C). The
plate was then dried for 2 hours in the air and 30 min on a heating plate (80 °C).
255
Publication IV
Development was carried out with CHCI3-MeOH-H20 (64:36:8) as mobile phase
and it was sprayed with 0.5 g thymol in 95 ml EtOH and 5 ml H2S04 (cone).
Cytotoxicity study using KB cell culture
The cytotoxicity of the compounds was determined using a KB cell line (ATCC CCL
17; human nasopharyngeal carcinoma). The test was carried out with some
modifications according to the screening technique of Swanson & Pezzuto (1990)
in 96-well plates (Falcon) with an inoculum of 2.5 x 104 cells/ml. Test solutions
were made as stocks in 20% ethanol in water. Before testing, the solutions were
diluted 20 fold and final ethanol concentration was 1% (v/v) or less. Total assay
volume was 150 uJ. For quantification of cytotoxicity 15 ul of an aqueous solution of
methylthiazolyltetrazolium chloride (MTT, Fluka) with 5 mg/ml PBS was added
(Mosmann, 1983). After incubation at 37 °C for 4 h, the metabolically active cells
produced an insoluble formazan dye. The medium was drawn off and the formazan
dye was dissolved using 150 ul of 10 % SDS (sodium dodecylsulfate) in water.
After 24 h of incubation at room temperature, the optical density was measured at
540 nm using a microplate reader (MRX, Dynex Technologies). For determination
of the IC50 values, the optical density was plotted against the log concentration.
The test was performed at least in duplicates.
Antibacterial activity
Antibacterial activity against B. cereus (ATCC 10702), S. epidermidls (ATCC
12228), M. luteus (ATCC 9341) and E. coli (ATCC 25922) were assessed using
the doubling dilution method (Liu et al., 1999).
Securigenin-3ß-0-ß-6-deoxyguloside (2)
White powder (11.1 mg); [a]24 -61.0° (MeOH, d.O); UV Amax (MeOH): 217 nm;
positive FAB-MS m/z 573 [M+Na]+, 551 [M+H]+, 443 [M+K-deoxyhexosyl]+, 369
[M+H-desoxyhexosyl-2H20]+; 1H NMR (500 MHz, CD3OD) Table 1; 13C NMR (75.5
MHz, CD3OD) Table 2.
256
Publication IV
Sarmentosigenin-3ß-0-ß-6-deoxyguloside (3)
White powder (9.1 mg); [a]24- 26.0° (MeOH, c 2.3); UV Amax (MeOH): 214 nm;
positive FAB-MS m/z 567 [M+H]+, 421 [M+H-deoxyhexosyl]+; 1H NMR (500 MHz,
CD3OD) Table 1; 13C NMR (75.5 MHz, CD3OD) Table 2.
19-Hydroxy-sarmentogenin-3ß-0-ß-6-deoxy-guloside (4)
White powder (23.4 mg); [a]24 - 36.0° (MeOH, c 1.0); UV Amax (MeOH): 214 nm;
positive FAB-MS m/z 553 [M+Hif; 1H NMR (500 MHz, CD3OD) Table 1; 13C NMR
(75.5 MHz, CD3OD) Table 2.
Sarmentogenin-3ß-0-[a-allosyl-(1 ->4)-ß-6-deoxyallosidel (5)
White-brown powder (39 mg); [a]24 - 5.2° (MeOH, c 2.3); UV Amax (MeOH):
214 nm; positive FAB-MS m/z 573 [M+K-H-hexosyl]+; 1H NMR (500 MHz, CD3OD)
Table 1 ; 13C NMR (75.5 MHz, CD3OD) Table 2.
Securigenin-3ß-0-[a-allosyl-(1 ->4)-ß-6-deoxyalloside] (6)
White-yellow powder (2.4 mg); [a]24 - 28.7° (MeOH, c 2.2); UV Amax (MeOH): 214
nm; positive FAB-MS m/z 573 [M+Na+H-hexosyl]*; 1H NMR (600 MHz, CD3OD)
Table 1 ; 13C NMR (75.5 MHz, CD3OD) Table 2.
3,15-Dihydroxy-18-norabieta-3,8,11,13-tetraene (7)
Yellow-brown powder (6.9 mg); [a]24 + 23.5° (MeOH, c 2.9); UV Amax (MeOH): 273
nm; positive EI-MS m/z 301 [M-H]T, 258, 216, 202, 188, 173, 149, 85, 83, 49; 1H
NMR (300 MHz, CD3OD) 8:1.04 (3H, s, H-20), 1.53 (1H, m, H-1b), 1.56 (3H, s, H~
17), 1.58 (3H,s, H-16), 1.62 (1H, m, H-6b), 1.93 (3H, brs, H-18), 2.23 (1H, m, H-5),
2.29 (1H, m, H-6a), 2.36 (1H, m, H-1a), 2.48 (2H, brd, J = 3.7 Hz, H-2), 2.70 (1H,
dd, J = 9.8, 18.9 Hz, H-7), 2.90 (1H, dd, J = 7.2, 18.4 Hz, H-7a), 6.79 (1H, d, J =
8.3, H-11), 6.91 (1H, d, J=8.3, H-12); 13C NMR (75.5 MHz, CD3OD) Table 2.
257
Publication IV
2,3,7-Trihydroxy-6-oxo-1,3,5(10),7-tetraene-24-nor-friedelane-29-oic acid
methylester (11)
Yellow powder (5.3 mg); [a]24 - 45.5° (MeOH, c 3.03); UV Amax (MeOH): 284, 321
nm; positive EI-MS m/z 496, 263, 248, 234, 203, 44; 1H NMR (300 MHz, CD3OD) cS:
0.69 (3H, s, H-27), 0.96 (1H, m, H-22b), 1.08 (3H, s, H-28), 1.15 (3H, s, H-30),
1.37 (3H, s, H-26), 1.43 (1H, m, H-21 b), 1.45 (1H, m, H-16b), 1.49 (3H, s, H-25),
1.59 (1H,d, J=7.8Hz, H-18), 1.66 (1 H, dt, J=4.1, 14.1 Hz, H-12b), 1.71 (1H, dd,
J =7.4, 14.7 Hz, H-19b), 1.82 (1H, m, H-12a), 1.85 (1H, m, H-15b), 1.91 (1H, m,
H-16a), 1.97 (1H, dt, J = 4.2, 14.2 Hz, H-11b), 2.12 (1H, dt, J = 4.1, 13.8 Hz, H-
22a), 2.16 (1H, m, H-11a), 2.17 (1H. m, H-21a), 2.47 (1H, brd, J = 15.5 Hz, H-
19a), 2.57 (3H, s, H-23), 2.88 (1H, m, H-15a), 3.55 (3H, s, H-31), 6.86 (1H, s, H-
1a); 13C NMR (75.5 MHz, CD3OD) Table 2.
Acknowledgements
The authors are very grateful to the healers, midwives and the inhabitants of
Chikindzonot, Ekpedz and Xcocmil, Yucatan (Mexico) for their collaboration, for
their friendship and hospitality during the fieldwork. The botanical identification was
performed in collaboration with the numerous specialists of the Centra de
Investigacion Cientifica de Yucatan (CICY) and the National Herbarium of Mexico
(MEXU). Particularly, we would like to thank Dr. I. Olmsted, Mr. J. Granados, Mr. P.
Simâ, Mr. J.C. Trejo, Dr. R. Durân of CICY as well as Dr. O. Tellez, Dr. R. Lira, Dr.
J. Villasehor and Dr. M. Sousa of MEXU. We are grateful to Prof. Dr. Brigitte Kopp,
Institute of Pharmacognosy, University of Vienna for the reference substances,
desglucocheirotoxol and strophanolosid. The authors thank Dr. O. Zerbe (ETH,
Department of Applied BioSciences) for assistance in NMR measurements, M.
Wasescha (ETH, Department of Applied BioSciences) for performing KB cell
assays, Dr. E. Zass (ETH, Department of Chemistry) for literature search, O.
Greter, R. Häfliger and Dr. W. Amrein (ETH, Department of Chemistry, MS-service)
for recording mass spectra. We are grateful to Prof. H. Budzikiewicz (University of
Cologne, Institute of Organic Chemistry) for performing the HRESI-MS spectra.
This research owes a lot to the help of Dr. Hongmei Liu (ETH, Department of
258
Publication IV
Applied BioSciences) and Dr. J. Orjala (Agra Quest Inc. Davis, USA). Financial
support by SDC (Swiss Agency for Development and Cooperation, Berne,
Switzerland) and the SANW (Swiss Academy of Natural Sciences) is gratefully
acknowledged.
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260
Conclusion
15 Conclusion
The healers and midwives of the three Yucatec Mayan communities, Chikindzonot,
Ekpedz and Xcocmil, have an extensive and deeply rooted knowledge about
medicinal plants. Forty informants reported a total of 360 plant species as
medicinal and 1828 individual-use reports were documented. The arrangement of
these responses into nine groups of uses formed the basis for an analysis of the
indigenous uses. The gastrointestinal problems are the most frequently mentioned
health problem of the study region, followed by skin problems. Illnesses according
to the Maya of Yucatan are classified as being humorally "hot" or "cold". However,
these characteristics were never spontaneously mentioned in the case of medicinal
plants. The healers first made references to the disease and its humoral
characteristic, and then referred to the herbal medicine indicated and its property.
In general, the humoral system is only one of the explanatory models used in this
medical system. Also form, texture or color of a plant and dreams about the healing
potency of a plant are important selection criteria.
The study of medicinal plants in comparison with non-medicinal ones showed that
taste and odor characteristics include considerable information about their use in
the treatment of illnesses. Aromatic plants are, for example, indicated to treat
diarrhea and vomiting. Taste and odor further help to distinguish between used and
non-used plants and play an important role in plant selection. Another interesting
point is that there was no difference in the percentage of species classified as bitter
between the medicinal plants and the non-medicinal ones. Thus bitterness is not a
specific critérium for plant remedies.
The evaluation of 48 important medicinal plants in various bioassays had the aim
to better understand the use of Yucatec Maya phytomedicines and their
pharmacological effects. Some correlations between the activities obtained in the
bioassays and the indigenous plant use could be shown. Other indigenous uses
261
Conclusion
cannot be explained based on the results of the pharmacological models. The
evaluation of these plants with respect to activity and cytotoxicity is a contribution
in their study of safety and efficacy. However further investigations should be made
this field.
For the detailed phytochemical study Crossopetalum gaumeri (Celastraceae) was
chosen due to its oral use against gastrointestinal problems, which is the most
frequent health problem in the study region. In addition, it is orally and locally used
in the treatment of snake bites. Furthermore, the extracts showed positive results
in different bioassays. The polar and nonpolar extracts of the roots of
Crossopetalum yielded six new compounds, four cardenolides and two terpenes.
Even though the cardenolide glycosides showed high cytotoxicity, they are not of
interest in the development of new anticancer drugs due to their strong effects on
the human heart and their small therapeutic window. The roots of C. gaumeri
showed good antibiotic and antiparasitic activity. However its oral use as a remedy
in gastrointestinal disorders must be viewed at with caution because of its high
cytotoxicity.
There is considerable knowledge about plant use by indigenous peoples and there
are established methods for testing the safety and efficacy of medicinal plants. The
aim of this ethnobotanical - phytochemical study was to forge a link between these
two areas. The study achieved this by investigating the activity of plants commonly
used by Yucatec Mayan healers and midwives. It is hoped that this collaborative
work between indigenous people and Western researchers has been mutually
beneficial and that it has helped to support the use of medicinal plants in the
region. Similarly, investigation of the phytochemistry and activity of these plants is
a further step towards the development of safer and efficient phytomedicine.
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List of publications
Ankli, A., Sticher, O., Heinrich, M., 1999. Medical Ethnobotany of the Yucatec
Maya: Healers' consensus as a quantitative criterion. Economic Botany 53,
144-160.
Ankli, A., Sticher, O., Heinrich, M., 1999. Yucatec Maya medicinal plants versus
nonmedicinal plants: Indigenous characterization and selection. Human
Ecology 27, 557-580.
Ankli, A., Heinrich, M., Bork, P., Wolfram, L., Bauerfeind, P., Brun, R., Schmid, C,
Weiss, C, Bruggisser, R., Gertsch, J., Wasescha, M., Sticher, O., (submitted).
Yucatec Mayan medicinal plants: Evaluation based on indigenous uses.
Journal of Ethnopharmacology.
Ankli, A., Heilmann, J., Sticher, 0., Heinrich, M., (accepted 2000). Cytotoxic
cardenolides and antibacterial terpenoids from Crossopetalum gaumeri.
Phytochemistry.
Heinrich, M., Ankli, A., Frei, B., Weimann, C, Sticher, O., 1998. Medicinal plants in
Mexico: Healers' consensus and cultural importance. Social Science and
Medicine 47, 1859-1871.
List of poster presentations
Ankli, A., Heneka, B., Orjala, J., Sticher, O., Heinrich, M., 1996. Plants in the
treatment of gastrointestinal disorders in two Yucatec Maya communities
(Mexico). Joint Meeting of the Society for Economic Botany and International
Society for Ethnopharmacology: Plants for food and medicine, London, UK, 1-
6 July. [Honorable mentioned]
Ankli, A., Heilmann, J., Sticher, O., Heinrich, M., 1999. New terpenoids from
Crossopetalum gaumeri- A Yucatec Maya medicinal plant. Joint Meeting of:
American Society of Pharmacognosy; Association Française pour
l'Enseignement et la Recherche en Pharmacognosie; Gesellschaft für
278
Arzneipflanzenforschung; Phytochemical Society of Europe. Amsterdam, The
Netherlands, 26-30 July.
Heinrich, M., Ankli, A., Frei, B., Weimann, C, Sticher, O., 1999. Arzneipflanzen in
Mexiko: Intra- und interkultureller Vergleich ethnnobotanischer Daten.
Deutsche Gesellschaft für Tropenökologie. Ulm, Germany, 17 February.
Oral presentations
Ankli, A., Sticher, O., Heinrich, M., 1997. Medicinal plants versus nonmedicinal
plants - Yucatec Maya selection criteria. II Congreso Intemacional
Etnobotânica '97. Mérida, Yucatan, Mexico. 12-17 October.
Ankli, A., Frei, B., Weiss, C, Heinrich, M., Sticher, O., 1997. Feldforschung:
Grenzen und Möglichkeiten. Arbeitstagung: "Rechte an biogenetischen
Ressourcen". Berne, Switzerland, 17 June.
Ankli, A., 1999. Charakterisierung der Medizinalpflanzen gegenüber den Nicht-
Medizinalpflanzen durch die Maya in Yucatan (Mexiko). Ethnologisches
Seminar der Universität Zürich. Ethnobotanik und Ethnomedizin in der
Basisgesundheitsversorgung. Zurich, Switzerland, 21 June.
279
Curriculum Vitae
1967 Born on June 25, Laufenburg, Switzerland
1974-1979 Primary school, Stein AG
1979-1983 Secondary school, Rheinfelden
1983-1987 Grammar school of Mathematics and Natural Science, Basel
Summer 1987 Language school in Saffron Waiden, UK
1987-1992 Study of Pharmacy at ETH Zurich
Diploma in Pharmacy (Eidgenössisches Staatsexamen)
1993-1994 Community pharmacist in Kreuz-Apotheke, Winterthur;
Language school in Cuemavaca, Mexico
1994-1995 PhD study: Ethnobotanical fieldwork in Yucatan, Mexico
1995-2000 PhD study under the supervision of Prof. Dr. 0. Sticher, at
section Pharmacognosy and Phytochemistry, Institute of
Pharmaceutical Sciences, ETH Zurich
Teaching in practical courses Pharmacognosy and
Phytochemistry I and II
1996-1999 Part-time employment as community pharmacist in Carmen-
Apotheke, Zurich
January 2000 Final examination to obtain the degree of Doctor of Natural
Sciences, ETH Zurich
280