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Flavonoids, Alkaloids and Tannins of Three Varieties of Horse Radish (Moringa Oleifera
Lam.) Extracts: Potential Components of Toothpastes
Jhoseph B. Carranza,Precious Gem L. Molina,John Joseph R. Ortañez, Lorcelie B. Taclan*,
Vicky C. Mergal and Orlex Yllano
Adventist University of the Philippines
Puting Kahoy, Silang, Cavite
Moringa oleifera Lam is a very common plant in tropical countries such as the
Philippines. Recently, the Philippine Congress approved it as the national vegetable because of its
various health benefits. The study generally identified the secondary compounds of the three
varieties namely; Native, Yard Long, and Chinese through phytochemical analysis. Specifically,
to establish the secondary compounds of the three varieties that would be potential components of
toothpaste. Gathering of samples of the three varieties was done at the university campus. Each
variety was dried using the Multi commodity heat pump dryer to preserve their nutritional
contents before crude extraction that was done at the Department of Science and Technology-
Industrial Technology Development Institute (DOST-ITDI) using 95% ethanol. Each extract was
subjected to phytochemical analysis using standard laboratory procedures. Results revealed that
the extracts contain flavonoids, alkaloids and tannins that are known to have antimicrobial and
anti-inflammatory properties, thus, these Moringa varieties have potential natural components in
the manufacture of toothpastes.
Keywords: Moringa oleifera L., flavanoids, alkaloids, tannins
Growing up in a tropical country, people have the tendency not to notice the beauty of the
vegetation that surrounds them. More often than not, the plants that are easily seen in our
backyard are taken for granted. Why say this? It is because the researchers are also guilty of it.
For example, Moringa oleifera Lam. or commonly called as malunggay in the Philippines, the
plant that is present in every Pinoy’s food, backyard, or sometimes in an abandoned lot. Not only
Moringa oleifera Lam. is easily seen in the Philippines, the country has the most researched
proven variety of the commodity. According to Nelly Favis-Villafuerte (2015), there are 13
different species of moringa plant and the best-known species and the most widely cultivated is
this specie native to the Philippines. The leaves, flowers, seeds, pods, roots, bark, gum, and the
seed oil from the malunggay plant are continually being subjected to intensive research and
development programs because the various constituents of the Moringa oleifera Lam. are known
to have, among other properties, anti-diabetic, anti-hypertensive, anti-inflammatory,
anti-microbial, anti-viral, anti-parasitic, anti-tumor and anti-aging activities. Though the plant is
native to the Philippines, it was also entitled as one of the world’s most useful plant (Marro,
2004).
The human’s oral cavity has many microorganisms inside, be it anaerobes or aerobic
bacteria. In fact, over 700 different kinds of bacteria have been detected in the human oral cavity.
The normal number of strains that can be found on a human oral cavity is 34 to 72 varieties only.
These are not harmful bacteria (Woudstra, 2015). Every part of the oral cavity has a different
concentration of microorganism. A study entitled Aerobic and Anaerobic bacteria in subgingival
and supragingival plaques of adult patients with periodontal disease compared the bacteria
present in plaques supragingival and subgingival. A total of 42 bacterial strains were isolated
from subgingival plaques, which of 24 (51.1%) belonged to 7 anaerobic species and 18 (42.9%)
to 12 aerobic species. There were more aerobic (33/53; 62.3%) than anaerobic bacteria (20/53;
37.7%) in supragingival plaques (Daniluk, 2006). These bacteria help in the first stage of food
digestion which is the breaking down or chewing of food into smaller pieces. The bacteria are
with the saliva that initiates the digestion and aid in chewing the food. The food then is made into
smaller, soft mass called bolus (YGOY Health Community, 2015). One of the most harmful
bacteria that can be found in the oral cavity is the Streptococcus mutans. It lives inside the mouth
and feeds on the sugars and starches that a person eats. That alone wouldn’t be so bad, but as a
by-product of its ravenous appetite, it produces enamel-eroding acids, which make Streptococcus
mutans the main cause of tooth decay in humans (Woudstra, 2015). Streptococcus mutans is an
indigenous micro flora of the oral cavity. It cannot be completely eliminated from the mouth and
the more it stays in the oral cavity, feeding on the starch and sugar that the person ingests, the
more they become in number that will be very bad and will result in greater probability of having
dental caries. Once a person gets a strain of oral bacteria, the person would not likely to get rid of
it but the good news is that people can control the bacteria living inside their mouth with good
oral hygiene. A study published in the Journal of Periodontology (JOP), the official publication of
the American Academy of Periodontology (AAP) demonstrates that flossing when made a routine
can actually help in reducing the amount of gum disease-causing bacteria that are found in the
oral cavity, thus giving people a better oral health. (American Academy of Periodontology, 2016)
Objectives of the Study
General Objective
To identify the secondary compounds of three varieties of Moringa oleifera Lam.
extracts namely; Native, Chinese, and Yard Long malunggay using phytochemical analysis.
Specific Objectives
1. To identify the appropriate solvent in the extraction of the secondary compounds of
native, yard long, and Chinese malunggay;
2. To identify the secondary compounds of the three varieties of malunggay that would be
potential components of toothpaste.
Review of Literature
Moringa oleifera Lam.
Malunggay in the Philippines is well known for its nutritional value as herbal medicine
(Fuglie, 2000). This plant is well cultivated in areas with tropical climate such as the Philippines,
India and Africa. This plant can grow to as high as 9meters with erect white trunks. The
compound leaf has about 3 to 9 leaflets. The plant has white fragrant flowers that produceslong
pods with 3-angled winged seeds (Medical Health Guide, 2011).
The Horse radish plant scientific classification:
Kingdom: Plantae
Division: Magnoliophyta
Class: Magnoliopsida
Order: Brassicales
Family: Moringaceae
Genus: Moringa
Species: Moringa oleifera Lam.
Different solvents used in the extraction of phenolic content of plants
Three genotypes of horseradish roots were collected in (latitude 57° 03’ N, longitude 22°
91’ E) during the period from September to November, 2011. For analyses, the average sample of
300 grams was collected by the researcher; this study took 3 unknown genome of roots. Each
genome of fresh roots were washed, peeled and homogenized (for 5 minutes). All samples of one
type of horseradish were homogenized together in order to obtain representative sample.
Figure 1 Total phenolic content of horseradish extracts
Reading of the Total Phenolic Content of horseradish extracts showed differences depending
on solvent used, namely: n-hexane (HE), ethyl acetate (EA), diethyl ether (DI), 2-propanol (PR),
acetone (AC), ethanol 95% (ET), ethanol/water/acetic acid (80/20/1/v/v/v) (EWA), and
ethanol/wataer (80/20 v/v) (EW). As the best solvents ethanol and ethanol / water solutions can
be chosen (Tomsone, 2012).
Effect of ethanol/water solvent on phenolic contents
A study revealed that phenolic compounds extracts were significantly dependent on the
concentration of ethanol/water solvent. The study revealed the highest phenolic content is
extracted when ethanol/water was used as a solvent compared to other solvents in the study
(Chunli, 2015).
Secondary Components of Moringa oleifera Lam. as an Anti-microbial
Flavonoids and alkaloids that are found in some plants is said to have an antiviral and
antimicrobial activities. A study done by the Faculty of Pharmacy, Department of Pharmaceutical
Microbiology, Gazi University, Ankara, Turkey proved that this was true. They tested the antiviral
activity of the compound by testing it against DNA virus herpes simplex type 1 and RNA virus
influenza (type-3). The Antibacterial activity was assessed against the following bacteria and their
isolated strains: Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis, Klebsiella
pneumoniae, Acinetobacter baumanni, Staphyloccocus aureus, Enterococus faecalis, and Bacillus
subtilis (Ozcelik, 2011). Adding to the wonders of flavonoids, a study by Salvatore Chirumbulo
stated that flavonoids has anti-inflammatory agents thus making it a great potential to be used as a
main ingredient in an anti-inflammatory drug (Chirumbulo, 2010). To prove the antibacterial
effect of Alkaloid, Shachi Singh et al did a study on several gram positive and gram negative
bacterial strains and arrived to a conclusion that the antibacterial test conducted revealed that
alkaloids have antibacterial property (Shachi et al, 2011). Phenolic compound on each plant
differs, therefore. it is hard to develop a standard extraction procedure suitable for the extraction
of all plant phenols. But, for the extraction of Moringa oleifera Lam., the use of ethanol and
ethanol/water (80/20 v/v) are the best solvents to use (Tomsone 2012).
Antibacterial Potential of Moringa oleifera Lam.
This present study is similar to that of Caurez (2012), the researcher evaluated the
antibacterial potential of Moringa oleifera Lam. root extracts in gram-positive (Staphylococcus
aureus and Bacillus subtilis) and gram-negative (Eschirichia coli and Salmonella typhii) bacteria.
The aqueous (distilled water) and alcohol (95% ethyl alcohol) root extracts of Moringa oleifera
Lam. exhibited antibacterial activity against S. Aureous and S. typhii. On table 1 is the
antibacterial acvitiy of Moringa Oleifera Lam. leaf juice extracts against some human pathogenic
bacteria.
Table 1 Antibacterial acvitiy of Moringa Oleifera Lam. leaf juice extracts against some human pathogenic bacteria (Caurez 2012)
Bacteria
Zone of Inhibition(mm)
Fresh leaf juice Aqueous extractsa
Aqueous extractsa
Ethanol Extractsa Positive Control
Tetracycline
Liquidb Powdera dissolved in DMSO
Fresh leaves Dried leaves Fresh
leaves Dried
Leaves
Cold Hot Cold Hot
Gra
m N
egat
ive
Shigella shinga 20.2±0.04 36.2± 0.08 7.75±0.56 + + + 17.5±0.34 + 12.20±0.13
Pseudomonas aeruginosa 17.00±0.66 39.60±0.49 15.00±034 + + + 21.21±0.05 + 18.30±0.12
Shigela sonnei 25.1±0.12 33.5±0.12 13.45±0.04 + + + 21.50±0.08 + 14.16±0.23
Pseudomonas spp. 25.2±0.04 42.3±0.16 27.5±0.21 + + + 21.25±0.1.3 + 20.16±0.19
Gra
m P
ositi
ve
Staphylococcus aureus 15.23±0.05 36.4±0.08 12.0±0.12 + + + + + 12.76±0.02
Bacillus cereus 22.4±0.28 29.25±0.2 8.00±0.42 + + + 16.25±0.04 + 9.34±1.3
Streptococcus-B-haemolytica 18.0±0.04 35.15±0.12 10.75±0.24 + + + + + 8.33±2.01
Bacillus subtilis 21.6±0.04 33.75±0.2 17.25±0.14 + + + 20.23±0.56 + 7.25±0.08
Sarcina lutea 18.1±0.04 34.4±0.44 8.50±0.09 + + + 19.50±0.21 + 8.66±0.13
Bacillus megaterium (Entero) 19.0±0.04 39.25±0.2 14.75±0.04 + + + 20.50±0.04 + 20.16±0.43
Values are presented as mean ± S.E. of triplicate experiments. + = Growth a Diameter of inhibition zone
including diameter of disc 6 mm (tested at a volume of 10 μl/disc at a concentration of 1175μg disc-1). b Diameter of inhibition zone including diameter of disc 6 mm (tested at a volume of 10 μl disc-1). DMSO = Dimethylsulfoxide
In the following aspects, the study used the extracts of Moringa oleifera Lam. leaves and the
organism tested is Streptococcus mutans. Philippine Council for Health Research and Development
published an article entitled “Moringa Potential Ingredient for Mouthwash/toothpaste”, the article
expounded that the study was conducted by researchers from Our Lady of Fatima University. In the study,
the result of the microbial in vitro test performed by the University of the Philippines Los Banos-National
Institute of Molecular Biology and Biotechnology (BIOTECH_UPLB) showed that juice of Moringa oleifera Lam. leaves inhibited the growth of Streptococcus mutans. Thus, the result of the present study
provided a framework for the researchers’ future study in the development of a Moringa oleifera Lam.
toothpaste for the inhibition of Streptococcus mutans in the oral cavity.
Components of a Basic Toothpaste
Dental Health Foundation in Ireland made a detailed and interesting article about the ingredients
of a basic toothpaste which is composed of abrasives, detergent, binding agents, humectants, flavoring
sweetening and coloring agents, preservatives, water, fluoride, and other therapeutic agents. Abrasive. Abrasive are the cleaning and polishing agents in a toothpaste and account for about a
third of the toothpaste by weight. Most abrasives are chalk or silica based. Examples are dicalcium
phosphate, sodium metaphosphate, calcium carbonate, silica, zirconium silicate or calcium pyrophosphate.
Abrasives differ in strength. An international standard defines a test paste against which toothpaste
abrasivity can be assessed. The unit of measurement is known as the Relative Dentine Abrasivity (RDA)
or the Relative Enamel Abrasivity (REA)
Detergent. Detergent (1-2%) foam and loosen plaque and other debris from the tooth surface.
Principal examples are sodium lauryl sulphate and sodium N-lauroyl sarcosinate.
Binding agents. Binding agents (1%), these agents prevent separation of solid and liquid
ingredients during storage. They are usually derived from celullose, sodium carboxy-methyl cellulose
being the most commonly used. Carrageenans (seaweed derived), xantham gums and alginates are also
used.
Humectants. Humectants (10-30%) acts to retain moisture and prevent the toothpaste from
hardening on exposure to air. Glycerol, sorbitol and propylene glycol are commonly used. Glycerol and
sorbitol also sweeten the toothpaste, though this is not their main function.
Flavoring, sweetening and coloring agents. (1-5%) Peppermint, spearmint, cinnamon,
wintergreen and menthol are among the many different flavoring used. While rare, mucosal irritations
from toothpaste (i.e., ulcerations, gingivitis, angular cheilitis, perioral dermatitis) are usually linked to
flavourings or preservatives they contain.
Preservatives. (0.05-0.5%) Alcohols, benzoates, formaldehyde and dichlorinated phenols are
added to prevent bacterial growth on the organic binders and humectants.
Fluoride and other therapeutic agents. The majority of toothpastes combine the caries
protection of fluoride with other therapeutic agents to control plaque, tartar and gum disease. The
inclusion of antibacterial agents can help individuals improve their plaque control. Mostly toothpastes
include triclosan, which has been shown to offer a clinically useful improvement in gum health. Other
pastes specially target “tartar” (hardened plaque) and use phyrosphosphate to inhibit the mineralisation of
dental plaque and hence the buildup of tartar (calculus). Toothpastes with desensitising agents are also
available for sensitive teeth. A study was conducted using an in vitro test of the effectiveness of
malunggay using disc diffusion and minimum inhibitory concentration (MIC) determination method
against human pathogenic bacteria. The study tested Shigella shinga, Pseudomonas aeruginosa, Shigella
sonnei, Pseudomonas spp, staphylococcus aureus, Bacillus cereus, Streptococcus-B- haemolytica, bacilus subtilis, sarcina lutea, Bacillus megaterium. The tests done using fresh leaf juice, powder from fresh leaf
juice, and cold-water extracts of fresh leaf exhibited inhibitory effect against all the tested Gram-negative
bacteria and Gram-positive bacteria and that includes Streptococcus-B-. However, the test using ethanol
extracts of fresh leaves gave a positive result on both Gram-negative and Gram-positive bacteria except in
S. Aureus and Streptococcus-B-haemolytica. The consequences of this investigation suggest that the
extracts and juice of M. oleifera Lam. can be used to discover antibacterial agent for developing new
pharmaceuticals to control studied human pathogenic bacteria responsible for severe illness” (Rhaman
2009). The related studies presented here were selected on the basis of their significance in promoting
directions for this present study. Streptococcus mutans is a good factor in the initiation of caries formation
(Simon 2007). The researcher expounded two pointers why Streptococcus mutans is a potent initiator of
caries. “Firstly, S. mutans is an anaerobic bacterium known to produce lactic acid as part of its
metabolism. Secondly, there is the ability of S. mutans to bind to tooth surfaces in the presence of sucrose
by the formation of water-insoluble glucans, a polysaccharide that aids in binding the bacterium to the
tooth” (Simon 2007). Also, unique to other species of microorganisms present in the oral cavity,
Streptococcus mutans does not slow down its metabolism even in a low pH environment. The effect of
flavonoids, alkaloids and tannins was established to have antimicrobial and anti-inflammatory effect by
previous studies, thus making it a potential component of toothpaste to reduce bacterial growth and
induce anti-inflammatory action.
Materials and Methods
Collection and preparation of Moringa oleifera Lam. for drying
The researchers gathered three kilograms of fresh Moringa oleifera Lam. for each variety to have
a yield of 300 grams dried of each. The researchers needed to have a minimum of 300 grams dried for
each variety in able to be tested for phytochemical analysis and antimicrobial susceptibility test. Weighing
scale was used to have an accurate measurement and to compute how much dried Moringa oleifera Lam.
the researcher can have in every certain weight of fresh Moringa oleifera Lam. that was subjected to
drying. Multi Commodity Heat Pump Dryer (MCHD) was used as the means of drying each variety; this
facility dehydrated the samples at an ideal drying condition which is 50°C. This equipment ensures the
embedded nutritional values, as well as the color of the samples will be retained after drying (Taclan,
2012). The researchers made sure that they got the desired dryness of all the varieties by weighing the
fresh varieties before drying. After 5h of drying, the researchers weigh the dried varieties and subjected
the dried varieties for another hour. After an hour, each variety was weighed again and when the present
weigh matched the weigh an hour before, it is an assurance that all varieties are dried enough and ready
for the next procedure. Heavy duty grinder was used to reduce the size of the dried varieties into smaller
particles or powdered form for the samples to be extracted.
Identification of the appropriate solvent in the extraction of secondary compounds
After preparing and having the needed weight of 300 grams each variety, the ethanolic crude
extraction of Moringa oleifera Lam. extracts, was done with the help of Department of Science and
Technology Industrial Technology Development Institute.
First, each powdered variety was soaked in the researcher’s chosen solvent which is 1.8 liters of
95% ethanol for 48 hrs. The use of rotary evaporator device was used after 48h of soaking to finally
extract the needed solution for the phytochemical analysis. There were still traces of the solvent used in
the crude extracts thus the use of 60°C water bath was needed to vaporize the remaining solvent as
another purification process.
Crude Extraction of Malunggay Leaves - Ground
Native Malunggay. The 300 grams of Native malunggay leaves produced 1.8 liters of
ethanolic extracts. Which was filtered and yielded 25 grams of semi solid crude extracts.
Yard Long Malunggay. The 300 grams of Yard Long malunggay leaves produced 1.8
liters of ethanolic extracts. Which was filtered and yielded 32 grams of semi solid crude extracts.
Chinese Malunggay. The 300 grams of Chinese malunggay leaves produced 1.8 liters of
ethanolic extracts. Which was filtered and yielded 35 grams of semi solid crude extracts.
Phytochemical analysis
The extracts of each variety were subjected to phytochemical analyses at the National
Institute of Molecular Biology and Biotechnology (BIOTECH) to identify their secondary
compounds which were Sterols, Triterpenes, Flavonoids, Alkaloids, Saponins, Glycosides, and
Tannins.
Bacterial Specimen
Only one microorganism strain was used in the investigation for antimicrobial assay
namely Streptococcus mutans. The microorganism was obtained from the PNCM Staff Philippine
National Collection of Microorganism. National Institute of Molecular Biology and
Biotechnology. University of the Philippines Los Baños College Laguna 4031 Philippines.
Antimicrobial Susceptibility Test
Procedure was done by PNCM Staff Philippine National Collection of Microorganism.
National Institute of Molecular Biology and Biotechnology (BIOECH). University of the
Philippines Los Baños College Laguna using 8mm diameter of cylinder cup.
Antimicrobial Susceptibility Test Interpretation
After 24 to 48h of incubation a ‘halo’ or ‘clearing’ around each disc will appear, this is
known as Zone of Inhibition. The plates will be inverted and a ruler will be used to measure the
diameter zone of inhibition, in millimeter. Results will be expressed in millimeter diameter zone
of inhibition. The size of the paper disc will be also recorded. A metric ruler will be placed across
the zone of inhibition, at the widest diameter, and the disk will be measured from one edge of the
zone to the other edge. The disc diameter will actually be part of that number. The results
acquired will be compared to the zone of inhibition of the standard antibiotic Phenol for bacterial
strain.
Figure 2
Flowchart of the Study
Collection and preparation for drying
of the three varieties of Moringa
oleifera Lam.
Preparation of Moringa
oleifera Lam. extract
Perform
Susceptibility test
on S. mutans
(Verification)
Phytochemical
Analysis
Analyses and
interpretation of
the results
Identification of extraction solvent
Results and Discussions
Concentration of 95% ethanol was the best solvent identified in the extraction of the three
varieties of malunggay
This study was focused on the identification of the secondary components of malunggay
compound. Thus, the researchers used ethanol as a solvent. Ethanol was proven to be the best
solvent for the extraction of secondary compounds of plants (Tomsone 2012).
Secondary compounds of three varieties of malunggay as potential components of
toothpaste
Table 2 shows the secondary components found in the three varieties of malunggay as
potential components of toothpaste. Native and yard long malunggay has an abundance (+++) of
tannins, this further explains that incorporating this on a toothpaste will give to it an inflammatory
agent reducing gingivitis. This result is congruent to the findings of Chirumbulo (2016). Out of
the three varieties considered, native malunggay has a moderate (++) amount of flavonoids.
Flavonoids was proven to have antibacterial and anti-inflammatory effect (Pietta 2000). All three
varieties of malunggay show traces (+) of alkaloids. Alkaloids have an antibacterial effect
(Kakhia 2012). The effect of flavonoids, alkaloids, and tannins was established by previous
studies, thus making it a potential component of a toothpaste to reduce bacterial growth and
induce anti-inflammatory action. (Table 2) Based on the result of the phytochemical analysis, it
was proven that Moringa oleifera Lam. extracts contains flavonoids, tannins and alkaloids that
made the three varieties of Moringa oleifera Lam. potential component of a toothpaste.
Table 2
Secondary components found in the three malunggay variety as potential components’ for a toothpaste
Sample Native Malunggay Chinese Malunggay Yard Long
Malunggay
Sterols (+++) (+) (+++)
Triterpenes (-) (++) (-)
Flavonoids (++) (+) (+)
Alkaloids (+) (+) (+)
Saponins (+) (++) (+++)
Glycosides (+) (+) (+)
Tannins (+++) (+) (+++)
Note: (+) Traces; (++) Moderate, (+++) Abundant (-) Absence of Constituents Reference:
Pharmacognosy, 15th edition, 2002, Trease and Evans
Present Constituents of Moringa oleifera Lam. Extracts
Sterols
Sterols, also known as steroid alcohols, are a class of chemicals that play multiple
important roles in the body. They have parts that can dissolve in fat-like molecules and parts that
can dissolve in water. The most widely known human sterol is cholesterol, which serves as a
precursor to steroid hormones and fat-soluble vitamins. Some people take plant sterols such as
vitamins A, D, E and K as supplements (Fox et al, 2015). Sterols, particularly cholesterol, are
integral to the stability of the cell membrane. The membrane is the outer covering of the cell,
analogous to the skin. Chemically, it is called lipid bilayer; lipids refer to fat-like molecules that
function to keep the environments inside and outside the cell separate, and bilayer means there
are two layers. The cholesterol embedded in the membrane maintains flexibility, fluidity, and
providing stability to the membrane over a range of temperatures. Sterols work to reduce the
absorption of cholesterol in the gut so more is lost in the feces. This in turn helps to lower total
cholesterol and LDL-cholesterol (bad cholesterol) in the blood (Main, 2015). A 2003 study in the
"American Journal of Clinical Nutrition" showed that sterols may interfere with the absorption of
excess cholesterol from the diet and therefore may be good for the heart and blood vessels.
Though potential to many health benefits, long-term effects and side effects of taking sterols still
need to be established. Consultation to a physician is highly recommended if planning to take
sterols (Fox et al, 2015).
Triterpenes
Originally synthesized by plants as metabolites, and are abundantly present in the plant
kingdom in the form of free acids or aglycones (Esser, 1999), triterpenes are synthesized via the
mevalonate pathway and are not regarded as essential for normal growth and development, and
although they do exist in plants in simple unmodified form, they often accumulate as conjugates
with carbohydrates and other macromolecules, most notably as triterpene glycosides. Triterpene
glycosides have important ecological and agronomic functions, contributing to pest and pathogen
resistance and to food quality in crop plants. They also have a wide range of commercial
applications in the food, cosmetics, pharmaceuticals, and industrial biotechnology sectors
(O’Maill et al, 2014). Though without scientific basis, in many Asian countries, herbal products
containing triterpenes are widely prescribed to prevent or treat a variety of diseases by the
traditional healers (Pharmar, 2013).
Flavonoids
Flavonoids are phenolic substances isolated from a wide range of vascular plants, with
over 8000 individual compounds known. They act in plants as antioxidants, antimicrobials,
photoreceptors, visual attractors, feeding repellants, and for light screening. Many studies have
suggested that flavonoids exhibit biological activities, including antiallergenic, antiviral,
anti-inflammatory, and vasodilating actions (Pietta 2000).
Alkaloids
Alkaloids are naturally occurring chemical compounds containing basic nitrogen atoms.
The name derives from the word alkaline and was used to describe any nitrogen-containing base.
Alkaloids are produced by a large variety of organisms, including bacteria, fungi, plants, and
animals and are part of the group of natural products (also called secondary metabolites). Many
alkaloids can be purified from crude extracts by acid - base extraction. Many alkaloids are toxic
to other organisms. They often have pharmacological effects and are used as medications, as
recreational drugs, or in entheogenic rituals (Kakhia, 2012). Antibiotic activities are common for
alkaloids and some are even used as antiseptics in medicine, e.g., berberine in ophthalmics and
sanguinarine in toothpastes, however, it is difficult to know the extent to which alkaloids give
antimicrobial protection in the plant (Margaret, 2013).
Saponins
Saponins are the second metabolites that are widely distributed in the plant kingdom. It
acts as a chemical barrier or shield in the plant defense system to counter pathogens and
herbivores (Cheok, 2014). Saponins are rich in pharmaceutical properties and recently many
studies focus on saponins' ability to increase immune responses, and possession of antibacterial,
antioxidant, anticancer, antidiabetic and anti-obesity properties (Cheok, 2014). A study proved
that 100% methanol fraction of saponin-rich extracts from guar meal exhibited antibacterial
activities against Staphylococcus aureus, Salmonella Typhimurium and Escherichia coli, however
the results showed 20% and 60% methanol fractions stimulated Lactobacillus spp. growth.
Aginoside saponins extracted from A. nigrum L. roots had significant antifungal activity.
Saponins isolated from seeds of Capsicum annum L. showed higher antimicrobial activity against
yeasts compared to common fungi. The n-butanol extracts of shallot basal plates and roots
exhibited antifungal activity against plant pathogenic fungi (Cheok, 2014).
Tannins
Tannins are considered as protectants against the growth of microorganisms such as
bacteria and fungi. These compounds affect microbial growth adversely and provide resistance to
the plants against them. Tannins act as redox buffers in plant cells, they protect the plant from the
deleterious effect of light and also impart antidessicant properties to the plant. A carbohydrate
core appears to play an important role in reducing the reactivity and solubility of tannins. It
enables them to be transported and stored in the plant without producing toxic effect (Pandey,
1993).
Glycosides
Glycosides are a class of molecules in which, a sugar molecule is bonded to a
"non-sugar" molecule. Glycosides play important roles in our lives. Many plants store
medicinally important chemicals in the form of inactive glycosides. The non-sugar portion
contains the biochemically active properties of medical interest. Once the glycoside is split into
its two components (sugar and non-sugar parts), the non-sugar component is now free to exert its
chemical effects on the body. For example, digitalis is a glycoside that when ingested, causes the
heart to contract (pump) more forcefully. This is useful in medicine, where heart failure is present.
Glycosides can be classified by the aglycone, glycone, or glycosidic bond. If the glycone portion
of a glycoside is glucose, then the researchers refer to the molecule as a glucoside. If the glycone
portion of the glycoside is fructose, then the researchers refer to the molecule as fructoside. If the
glycone portion of the glycoside is glururonic acid then the researchers refer to the glycoside as
a glucuronide. In the human, toxic substances are often bonded to glucuronic acid. The
glucuronic acid functions to increase the water solubility of the glycoside, which can be more
easily secreted. (http://www.nutriology.com/glycoside.html)
Comparison and Discussion of Different Solvents for Isolation of Phenolic Compounds
Phenolic content of plant extracts are affected by different factors such as, variety of
plant used, climate where the plant was taken, storage of the plant before and after extraction,
handling and processing during testing etc. Plants grown in different part of the globe was proven
to have different Total Phenolic Content. The recovery of polyphenols from plant materials is
influenced by the solubility of the phenolic compounds in the solvent used for the extraction
process. The solvents listed below are arranged in their unpolarity starting from the most unpolar.
Solvent polarity plays a key role in increasing phenolic solubility. It was proven that Total
Phenolic Count generally increased by increasing a polarity of solvents. Phenolic compounds on
each plant also differs, therefore, it is hard to develop a standard extraction procedure suitable for
the extraction of all plant phenols (Tomsone 2012).
Summary and Recommendations
Summary
The plant constituents analyzed were Sterols, Triterpenes, Flavonoids, Alkaloids,
Saponins, Glycosides, and Tannins. Native and Yard Long malunggay have a close presence of
components in general yet have a slight difference between Saponins. Yard long have an abundant
constituent while Native malunggay have only traces and Flavonoids; wherein Native malunggay
have moderate constituents while Yard long malunggay only have traces. Chinese malunggay
have the least presence of components in general. Even with different concentrations, all three
varieties have presence of flavonoids, tannins, and alkaloids, which are potential antimicrobial
and an anti-inflammatory agent. The crude extracts were tested for its anti-microbial property on
Streptococcus mutans through antimicrobial susceptibility test done at National Institute of
Molecular Biology and Biotechnology. All three varieties with their different concentrations
resulted negative to the said test. Based on the researcher’s gathering of related literatures,
ethanol is the best solvent in extracting phenol rich plants such as the Moringa oleifera Lam.
Conclusions
The study showed that: (1) Ethanol was appropriate for the extraction of the three varieties of
malunggay namely: Native, Yard-long and Chinese and; (2) based on the result of the
phytochemical analysis, it was determined that Moringa oleifera Lam. extracts contains
flavonoids, tannins and alkaloid which is a potential toothpaste component.
Recommendations
1. Future studies using other solvents for extraction may be conducted on same varieties of
malunggay.
2. Extraction method using mortar and pestle is recommended.
3. Based from the results of the phytochemical analysis, toothpaste with ground malunggay
may be developed.
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