A search for hepatoprotective agents of natural originshodhganga.inflibnet.ac.in/bitstream/10603/80883/10/10...A search for hepatoprotective agents of natural origin Department of

A search for hepatoprotective agents of natural origin

Department of Chemical Technology, Dr.Babasaheb Ambedkar Marathwada University, Aurangabad

26

In any phytochemical investigation of medicinal plants, their botanical

identification forms the most crucial and basic as any erroneous identity

here will render all further studies on chemistry, pharmacology and clinical

trial irrelevant.

3.1 Berberis aristata DC. Var. aristata Fam:- Berberidaceae

Regional and Other Names

Ben.: Daruharidra, Darhaldi; Eng: Indian Berberry ; Guj.: Daruharidra,

Daruhuladur; Hind.: Daruhaldi, Kashmal, Rasaut, Daruhald, Darhald,

Kashmar, Rasvat, Chitra, Chotra; Kan.: Maradarishana, maradarishina,

Daruhaldi, Doodamaradarisina, Mal.: Maramanjal, Maradrisina,

Kasturimanjal; Mar.: Daruhalad; Ori,: Daruharidra, Daruhaldi; Punj :

Sumulu, Chitra, Kasmal, Simlu; Tam.: Gangeti, Varatiu manjal,

Maramanjal, Mullukala, Usikkala, Kasturimanjal; Tel.: Manupasupu,

Kasturipaspu, Daruharidra.

Habit and Habitat

An erect, spinescent shrub, found in Himalayas from Garhwal to Bhutan, at

altitude of 1800-3000m.

Ayurvedic Description

Sanskrit name : Daruharidra, Rasanjana (dry extract)

Synonyms : Darunisa, Darvi, Pacapaca, Katamkatri, Parjanya,

Paita, Pitadaru

Properties : Rasa: Katu, tikta; Guna: Ruksa;

Virya: Usna, Vipaka.

Actions : Kaphapittahara, lekhaniya, sodhani, kanduhara,

vranadosahara, varnya, chedana, romasanjanana



27

Therapeutic uses : Karnaruk, netraruk, mukharuk, kandu, vrana,

meharoga, tvagdosa, sotha, Rasanjana Vranadosa

and putikarna.

Therapeutic Uses Mentioned in Ayurvedic Pharmacopoeia

The stem is used in amatisara (diarrhoea), medoroga (obesity),

urustambha (stillness/loss of movement of legs), kapharoga (diseases due to

excessive phlegm), karnaroga (diseases of the ear), mukharoga (diseases of

oral cavity), netraroga (ophthalmic diseases), kandu (itching), vrana (wounds)

and meha (diabetes).1

Properties and Uses

The drug ‘rasaut’ is regarded as bitter tonic and is reported to be used

as a cholagogue, stomachic, laxative, diaphoretic, antipyretic and antiseptic.

It is administered externally in painful eye affections, indolent ulcers and

haemorrhoids.

The fresh berries are laxative and antiscorbutic, useful in piles, sores

and eye disease particularly conjunctivitis. Mixed with the bark of

Cinnamonum tamala and honey, they are prescribed in leucorrhoea. A

decoction is used as mouth was for treating swollen gums and toothache.

Decoction of root bark is used in malarial fever.2

Ethanobotanical Studies

The Plant is used as antiulcerogenic, antidiabetic, antidysentric, in

ophthalmic, skin diseases3, as antidiarrhoeal, in menorrhagia, jaundice and

as alterative, astringent, deobstruent, diaphoretic and antiperiodic. The fruits

are used as laxative in leucorrhoea, swelling of gums and toothache and in

haemorrhoids whereas berries and laxative and antiscorbuiti. The bark is



28

used as an antimalarial. The root is used in rheumatism, ophthalmic

diseases, piles, menorrhagia, blood coagulation and chronic ulcers. Also it is

used as an antipyretic, antiulcerogenic, laxative, tonic; antimalarial;

antidiarrhoeal, in skin diseases, menorrhagia, jaundice, haemorrhoids and

as liver tonic. The root bark is employed in ophthalmic diseases, skin

diseases and boils haemorrhoids, to wash ulcers and wounds4 in liver

complaints, as antidiarrhoeal, in ear troubles backache and as a general

tonic.5

PHARMACOGNOSTIC STUDIES

The dried root under ordinary light is brown in whole form, faint

yellow in fractured form and greyish brown in powder form. Under ultraviolet

light the root appeared brown in whole form, greenish yellow in fractured

form and deep green in powder form.6

A study on the calcium oxalate crystals of forty five medicinally

important barks revealed that the bark contained solitary square rectangular

prism shaped calcium oxalate crystals in the cortex region. Under normal

light the bark powder appeared dark blond and under ultraviolet light

greyish green in colour.7

CHEMICAL STUDIES

The flowers were reported to yield five polyphenolic compounds,

namely E-caffeic acid, quercetin, chlorogenic acid, meratin and rutin.8The

bark afforded berberine chloride, palmatine chloride and a mixture of

palamatine and bebrine chlorides.9

The root showed presence of alkaloids, flavonoids, glycosides,

saponins and sterol and absence of terpenoids10. Another study, however,



29

PLATE NO.1



30

reported the presence of alkaloids and tannins in the root but flavonoids and

saponinis were reported to be absent.11

‘Rasut’ which is a brown almost solid extract prepared from wood or

root of Barbaris sp. was chemically assayed for obtaining an idea of its purity

and gross adulteration. Out of fifteen samples collected from the market,

seven either did not contain alkaloid or only traces of it. The variation in the

alkaloidal content in the rest of the samples was from 1.67 to 4.26 %.12

High performance thin layer chromatography and spectrophotometric

methods were described for the estimation of berberine from the root and the

average content reported by these methods was 2.76 % and 2.58 %,

respectively. The sensitivity by HPTLC method was 0.01 mg/ml and that of

spectroscopic method was 0.02 mg/ml 13. General derivatives of berberine

were prepared from dihydroberberine, 8-acetonyldihydroberberine and 8-

benzyldihydroberberine by making use of their enamine character and

evaluated for their biological activities .14

PHARMACOLOGICAL AND BIOLOGICAL STUDIES

Antiarthritic: An emulsion prepared from the gum on oral

administration (1 ml) failed to show any significant beneficial effect in

formalin-induced arthritis in albino rats.15

Heparin neutralizing Action: Berberine (1-3mg) neutralized in vitro the

anticoagulant action of 50 i.u. of heparin/ml on dog and human blood.

Larger dose (10mg/ml) had a paradoxical anticoagulant action. The activity

was found similar to that produced by protamine sulphate and toluidine

blue.16



31

General Pharmacology

The alcoholic extract showed moderate direct action on the intestine of

rabbit and uterus of guinea pigs, while action was observed on heart and

rectus abdominus of frog. Its antagonism with barium chloride was found to

be negligible, no action was observed with histamine.17

The alcoholic extract of the root was studied on the platelet activating

factor (PAF) on rabbit platelets. It inhibited the PAF-induced aggregation of

platelets in a dose dependent manner. It also inhibited the [3H]-PAF binding

to rabbit platelets in a competitive manner.18

In a preliminary study, the 50 % ethanolic extract of root showed

hypoglycaemic activity in rats, effects on CVS in dogs/cats and anticancer

activity against human epidermoid carcinoma of nasopharynx in tissue

culture. The extract was devoid of antibacterial, antifungal, antiprotozoal,

anthelmintic, antiviral, antispirochaetal and antifertility activities and effects

on CNS, isolated guinea pig ileum and rat uterus. LD50 value of the extract

was found to be 20 mg/kg i.p. in mice.19

Antimicrobial

Alcoholic extract of wood powder was found to be active

Staphylococcus aureus and Salmonella typhosa, while water extract did not

show any activity against Salmonella typhosa but was active against

Staphylococcus aureus.20

The methanolic extract of rhizomes did not show antibacterial activity

against Escherichia coli in vitro studies. The extract of tap root showed

antibacterial activity against Shigella flexneri type IV while it was inactive

against all species of Salmonella and Escherichia coli.21



32

The aqueous, hexane and alcoholic extracts of root when screened for

antimicrobial activity against bacillus subtilis, Escherichia coli, Proteus

vulgaris, Salmonella typhimurium, Pseudomonas aeruginose and

Staphylococcus aureus did not show any activity.22

Insecticidal

The alcoholic extract of root exhibited very mild/negligible inserticidal

activity against houseflies Musca domestica nebulo and mosquitoes Aedes

aegypti.23

The 50 % ethanolic extract of root, when screened for preliminary

biological activities showed effects of CVS in dogs/cats, isolated guinea pig

ileum and possessed confirmed anticancer activity against human

epidermoid carcinoma of nasopharynx in tissue culture. The extract did not

show the other activities tested viz. antibacterial, antifungal, antiprotozoal,

anthelmintic, antiviral, antispirochaetal, antifertility and hypoglycaemic

activities and effects on CNS. The MTD value of the extract was found to be

250 mg/kg i.p. in mice. In another study, the 50 % ethanolic extract of the

plant ( excluding root), exhibited CVS effects in dogs/cats and weak diuretic

activity in rats, while the extract was devoid of other activities viz.,

antibacterial, antifungal, antiprotozoal and antiviral activities and effects on

CNS in experimental animals and on isolated guinea pig ileum. LD50 of the

extract to be 316 mg/kg i.p. in mice. 19

Antimicrobial

Different extracts from stem bark were tested for their antimicrobial

activity against Staphylococcus aureus, enterococus faecalis, Escherichia coli,

Klebsiella pneumoniae, Enterobacter sp., Citrobacter sp., Serratina



33

marcescens, Pseudomonas aeruginosa, Proteus vulgaris, Premirabilis,

Edwardsiella tanda, Salmonella paratyphi A, Shigella dysentriae, Shigella

flexneri, Vibrio cholera, Vib parahaemolyticus, Plesiomonas shigelloides,

Aeromonas hydrophylla and Candida albicans. The crude aqueous extract

exhibited a zone of inhibition against 11 out of 20 microorganisms. The

gram positive bacteria Staphylococcus aureus and Enterococcus faecalis

were observed sensitive. Among the gram negative bacteria, Escherichia coli,

Klebsiella pneumoniae, Serratia marcescens, Proteus mirabills, Prvulgaris,

Salmonella paratyphi A, Shigella dysenteriae and Pseudomonas aeruginosa

exhibited a zone of inhibition. The alcoholic extract was found to be the best

for Candida albicans. Alkaloid fractions derived from the plant showed

stronger activity than pure berberine iodide.24

CLINICAL STUDIES

Antidiarrhoeal

Clinical studies with berberine were conducted in 356 patients of

cholera and compared with 264 patients treated with chloramphenicol at

Chittaranjan Hospital, Kolkata. Berberine was found to be effective in both

bacteriologically positive and negative patients. It reduced the mortality rate,

volume and duration of diarrhoea, intake of intravenous fluid and the

convalescence period. Berberine was found to be better than

chloramphenicol in this respect. It showed no side effect of toxicity in the

doses tried. Initial dose of berberine hydrochloride was 15 mg (oral), 6 hourly

first 2days, 8 hourly on the 3rd day and only one dose on the 4th day. Later

the doses were increased to 50 mg on admission, 50 mg twice daily till the 4th



34

to 5th day. The possible mechanism of berberine in cholera was proposed to

be its neutralizing action on the diarrhoea producing component.25

In a clinical study at Kasturba Hospital, Mumbai, berberine

hydrochloride was tried in 100 cases of gastroenteritis for its antidiarrhoeal

action and compared with equal number of controls. The drug was given

along with routine line of treatment. The recovery in berberine treated groups

was faster and quicker. There was no death in the berberine treated group,

while there were five deaths in the control group treated with bismuth,

kaolin, chloramphenicol and streptomycin. Further, berberine was tried

along as an antidiarrhoeal drug in additional 30 cases of gastroenteritis. It

arrested diarrhoea in all these patients with no mortality and toxicity.26

Berberine hydrochloride along with uneomycin compound in a dose of

1 ml twice daily on the first day and once daily on second day was found to

be most effective when treated for infective diarrhoeas of infancy as

compared to the patients treated with uneomycin compound and berberine

hydrochloride along.27

In a trial at Cama and Albless Hospital, Mumbai involving 200

children suffering from diarrhoea of various etiologies, berberine tannate was

assessed for its antidiarrhoeal effect and compared with equal number of

patients receiving chloramphenicol. It was found to be effective. Children

receiving berberine tannate recovered faster than the chloramphenicol group.

Only in about 3 per cent of the cases, the diarrhoea was not controlled in

berberine treated group. In 25per cent of the cases the diarrhoea was

controlled within 24-72h. The drug and probably some antispasmodic and

astringent action on the intestinal mucosa with no side effects.28



35

Berberine tannate (25mg/6hrs.) was clinically tried at Isolation

Hospital, Jaipur, in 65 children below 5 years with acute diarrhoea for its

antidiarrhoeal action and compared with an equal number of controls on

standard antidiarrhoeal drugs. The recovery in berberine treated children

was faster. It was found effective against diarrhoeas caused by Escherichia

coli, Shigella sp., Salmonella paratyphi, Klebsiella aerogenes and Faecalis

aerogenes.29

The effect of berberine was studied in the treatment of diarrhoea of

infancy and childhood. It was tried in 50 cases of gastroenteritis in children

at JLN Hospital, Ajmer and revealed beneficial effect particularly in mild (4-6

loose stools in 24h) and moderate cases (with 6-15 loose stools in 24h with

dehydration). In three cases, commuting led to the withdrawal of the drug.30

In another study at M.Y. Hospital, Indore the efficacy of bereberine

hydrochloride was studied in cases of acute gastroenteritis, acute bacillary

dysentery and acute amoebic dysentery alone or in combination with

standard antidiarrhoeal drugs. 129 cases of acute diarrhoeal disorders were

selected for the trial which consisted of 57 cases of acute enteritis, 37 cases

of acute bacillary dysentery and 35 cases of acute amoebic dysentery.

Berberine hydrochloride give orally in doses of 100 mg. t.i.d. showed curative

action in acute gastroenteritis, acute bacillary dysentery and acute amoebic

dysentery. Berberine hydrochloride when given in combination with

chloroampenicol and streptomycin had better curative action in cases of

acute gastroenteritis and acute bacillary dysentery than berberine

hydrochloride alone or chloroamphenicol and streptomycin in combination.

Combination of berberine hydrochloride with iodochlorhydroxyquinoline and



36

sulphadimine showed better curative action in acute amoebic dysentery than

any of the drug given alone. Berberine hydrochloride showed antispasmodic

effect in addition to its antidiarrhoeal action.31

Antigiardiasis

Twenty five patients (between the ages 1-10 years) of giardiasis were

treated at AIIMS, New Delhi with berberine in a dose of 5 mg/kg/day for

6days, and the results were compared with those of metronidazole given in a

dose of 10 mg/kg/day in nine patients. Twenty patients receiving vitamine B

complex syrup for 6days served as the placebo controls. Relief of clinical

symptoms was observed in twelve patients receiving berberine, three

receiving metronidazole and three receiving vitamine B complex. The stool

became free of giardia in seventeen patients receiving berberine, six receiving

metronidazole and five receiving vitamin B complex after 6days of

treatment.32

Trachoma Lesion

Patients with clinically active trachoma lesion (stage I and II) were

treated for 8 wk with eye drops of either 0.2 % berberine or 20 %

sulphacetamide or a combination of both. There was marked clinical

improvement in all the patients treated. The sulphacetamide eye drops gave

the best clinical results but the conjunctival scrapings of these patients were

invariably positive for inclusion bodies of Chlamydia trachomatis and there

infection.33

Antimalarial

The barberine sulphate in a doses of 3 to 7 grains t.i.d. on three

consecutive days failed to arrest paroxysms of malarial fever and reduced



37

number of malarial parasites present in the blood. In nine cases of malaria at

Carmichael Hospital, Kolkata for Tropical Diseases, in which barberine

sulphate was tried, in no instance was there any change in the signs and

symptoms produced by the disease.2

Oriental Sores

‘Rasaut’ on continuous local application was claimed to have carrying

degree of beneficial effect in some cases of oriental sores34. Infilteration of

beberine sulphate was reported to have beneficial effect in oriental sores.35

Berberine sulphate inhibited the growth of Leishmania tropica in vitro

studies. Local injection (1 % solution of Berberine sulphate was reported to

cure oriental sources within 2 or 3 wk. Berberine hydrochloride (2 %

solution) was used in 54 cases of oriental sore which were positive for

Leishmania tropica and devoid of secondary infection. Cure rate was 67 %

with an average cure rate time of about 4 wk. Face lesions showed marked

inflammatory reaction and slow response to treatment. The drug was

otherwise well tolerated and found safe.36

3.2. Acacia nilotica Linn. Fam.:- Mimosaceae


Ben. : Babla, Babul; Eng.: Babul, Black Babool, Indian Gum Arabic tree;

Guj.: Babaria, Baval, Kaloabaval; Hind.: Babul Kikar; Kan.:Gobbli, Karijali;

Mal.:Karivelan, Karuvelum; Mar.: Babhul, Vedibabul, Babhula; Ori.:

Bambuda, Baubra; Punj.:Sak; Tam:Karuvelamaram, Karrivelei, Karuvael,

Karuvelam; Tel.: Nallatumma, Tumma, Tuma.



38

Habit and Habitat 37

A moderate-sized tree with a spreading crown and feathery foliage found

throughout the drier parts of India.

Ayurvedic Description 37

Sanskrit name : Babbula

Synonyms : Barbura, Kinkirata, Yugmakanta, Suksmapatra,

Pitapuspa.

Properties : Rasa: Kasaya; Guna: Guru, ruksa; Virya: Sita;

Vipaka: Katu.

Actions : General : Kaphara, grahi, visaghna

Gum : Grahi, vrsya, sonitasthapana

Fruit : Stambhana, asthisandhanakara

Therapeutic uses : General : Kasa, amatisara, raktatisara, krmi,

kustha, raktapitta

Gum : Raktatisara, meharoga, pradara,

bhagna

Flower : Netraroga, prameha

Properties and Uses

The gum obtained from the tree is useful in diarrhoea, dysentery,

diabetes mellitus and sore throat. The extract of the gum is styptic, tonic and

astringent. The bark is reported to be astringent, demulcent and aphrodisiac.

The pods are expectorant.

Ethnobotanical Studies

The parts of the plant find use in diabetes, skin diseases and

leucorrhoea38. These are also used as an antidiarrhoeal, antidysentric39. The



39

stem bark is demulcent40 used in diabetes as astringent41, anthelmintic, in

skin diseases, cough and bleeding piles gonorrhea42 and as an

antiasthmatic43. The tender twigs are used as toothbrushes44 while the

thorns are used for joint pains and in oral cavity lesions. The seeds and

leaves extracts are used for general body vigour45. The leaves are used in

diarrhoea46, dysentery47 in headaches48, eczema49, abscess and ophthalmic

disorders50. The root is used for wound healing51 and for burning sensation.52

PHARMACOGNOSTIC STUDIES

The gum of the plant as such and in fractionated form appears golden

yellow to brown under ordinary light while it appears lemon yellow with

bright white patches under ultraviolet light. In the powdered form the gum

appears brownish yellow and light yellow, respectively under ordinary and

ultraviolet light.53

Gum arabic has been identified as one of the constituents of a Unani

compound drug preparation ‘hab-e-bukhar’, specially given in fever due to

elephantiasis and malaria.54

Chemical Studies:-

The acetone extract of the flowers contained stearic acid along with

three phenolic components viz. kaempferol-3-glucoside, isoquercetin and

leucocyanidin55. The pods showed the presence of steroids/terpenoids,

tannins, phenolics and saponins but were devoid of alkaloids and

flavonoids56. The seeds contained fatty oil which showed the presence of

capric, lauric, myristic, palmitic, stearic, oleic, linoleic, linolenic and

arachidic acids. Trace quantities of epoxy and hydroxy fatty acids were also

detected.57, 58



40

The shoot and seeds in preliminary chemical studies showed the

presence of saponinis, flavonoids and alkaloids59,60. The seeds in addition

contained many amino acids. The total essential amino acid content was

30.37g. Other constituents were also quantitatively analyzed. These included

ether extractive, crude protein, fibre, total ash, calcium, phosphrous, iron,

niacin and ascorbic acid.61

The free vitamin C and carotene content of the leaves,62 crude protein,

hemicellulose and cellulose were quantitatively estimated. The total ash,

silica, dry matter and neutral as well as acid detergent fibre contents were

also determined63. The leaves contained alkaloids and saponins64. A large

variation was seen in the chemical constituents of the leaves.65

Several polyphenolic compounds were reported from the bark which

were identified as quercetin, gallic acid, (+)-catechin, (-)epicatechin, (+)

dicatechin and (+)- leucocyanidin gallate66. Bark was also reported to contain

(-)-epi-gallocatechin67. The total water soluble extractives and tannin content

in the bark of trunk as well as the branches were found to be 18.44 % and

11.02 % and 27.36 % and 14 %, respectively.68

Phytochemical analysis of the polyphenolic complex present in the

bark of the plant revealed the presence of phenols, flavones, tannins and

glycosides. The total phenolic and tannin contents in the dried bark were

found to be 9.86 %, respectively; while the total flavonols and total O-

dihydric phenols including chlorogenic acid were 0.0025 % and 0.013 %

respectively. The bark contained asponins.64



41

The benzene extract of the root bark of the plant was found to contain

octacosanol, β-amyrin, β-sitosterol, betulin while the petroleum ether extract

of the root heartwood contained β-amyric and β-sitosterol.69

The detected mineral elements in the plant were calcium, copper, iron,

magnesium, zinc, potassium, sodium, aluminium, manganese and cerium.70

Pharmacological Studies:

Antidiabetic

The bark in the form of decoction (20 ml/kg) as well as the standard

drug tolbutamide produced a significant reduction in blood glucose levels in

mild alloxonised diabetic rabbits fasted for 18h. The maximum reduction was

2h after the administration of the plant extract in rabbits as compared to the

controls.71

The three leuminous seeds viz., A. benthami, A. modesta and A.

nilotica ssp. indica fed for one week were found to exhibit hypoglycaemic

effect (blood sugar lowered by 29.99 %, 27.68 % and 25.05 %, respectively) in

normal rats, but did not show any significant hypoglycaemic effect in

alloxanised diabetic rats (blood sugar lowered by 3.69 %, 2.24 % and 2.14 %,

respectively). The hypoglycaemic effect of these legumes was due to their

direct or indirect stimulation of β-cells of islets of langerhans to secrete more

insulin. In alloxanised rats, since β-cells of langerhans were destroyed there

was no hypoglycaemic effect observed when the animals were fed on

leguminous seed diets.72

Antimutagenic

The methanolic extract of the bark decreased the UV-induced

mutagenicity using the Escherichia coli WP-2 in a dose of 5 mg/plate. This



42

decrease might be due to some enzymatic action which reverted the

formation of pyrimidine dimmers.73

Antiproteolytic

Inhibition of total proteolytic (caseinolytic), tryptic (by hydrolysis of

benzoyl arginine p-nitroanilide) and chymotryptic (by hydrolysis of acetyl

ethyl ester) activities by ten species of legume seeds on human and bovine

pancreatic proteases were studied. Acacia seed extracts displayed more

pronounced action on human tryspin and chymotrysin. It was more effective

in inhibiting the total proteolytic activity of the bovine system.74

Antifertility

The aqueous extract of the flowers showed 11.5 % abortifacient

activity in rats. It was further screened for teratological abnormalities in

failure cases (where pregnancy was not prevented) in pregnant rats. The

fetuses showed gross external morphological and skeletal defects. The extract

of the stem bark at 2 % concentration revealed semen coagulant activity in a

preliminary screening.75

Nutritional Value

Nutritional evalution of the refined seed oil was done by rat bioassay

and using peanut oil as control. The animals fed on 10 % seed oil diet

showed poor growth performance and low feed efficiency ratio. The

digestibility of the seed oil was 90 % as compared to 94 % for peanut oil. The

seed oil in the diet of rats for 4wk did not product any abnormal serum lipids

or histopathological findings. The seed oil was apparently non toxic.58 The

deoiled seed cake contained 21.9 % protein and balanced amino acids but

also contained antinutritional factors, tannins (4.2 %) and saponins (2.4 %).



43

The nutrient and amino acid composition of the detoxified seed meal (PAM)

was almost similar to that of unprocessed seed meal except for

antinutritional factors. PAM was nutritionally evaluated using rat bioassay

procedure in a comparative study with cesain as standard. Nutritional

indices, biochemical parameters and histopathological findings indicated the

possibility of using PAM as supplementary feed for livestock animals.76


The 50 % ethanolic extract of the stem bark in a preliminary biological

screening exhibited antiprotozoal activity against Entamoeba histolytica, CVS

effect in dog/cat, antispasmodic activity in guinea pig ileum and CNS

depressant activity as evidenced by amphetamine hyperactivity test in mice.

The extract was devoid of antibacterial, antifungal, antiviral, hypoglycaemic

and anticancer activities. The LD50 was found to be 500 mg/kg i.p. in mice77.

A quanternary base picrate, (mp 242-44oC) isolated from 11 species

including the stem bark of the plant was reported to be pharmacologically

identical to choline78. The saline extract of the pollen grains stimulated the

ileum of guinea pig which was blocked by mepyramine and atropine. The

petroleum ether extract stimulated the rat uterus and the heart of pila which

was blocked by 2-bromo LSD. The effect of acid treated acetone extract was

blocked by mepyamine.79

Antimicrobial & Antibacterial

Air dried and powdered alcoholic and water extracts of the bark

exhibited significant in vitro antibacterial activity against Streptococcus

pyogenes, Staphylococcus aureus, Escherichia coil, Salmonella

typhimurium, Pseudomonas aeruginosa and Klebsiella sp. using the disc



44

method. Pseudomonas aeruginosa was found to be resistant to both the

extracts. Further, both the extracts were highly inhibitory to the gram

positive organisms in comparison with gram negative organisms tested. All

the microorganisms showed resistance against the petroleum extract.80

The alcoholic extract of the gum, leaf and fruit revealed in vitro

antibacterial activity against Staphylococcus aureus (zone of inhibition 10-19

mm) using agar diffusion method while it was devoid of any activity against

the other bacteria Bacillus subtilis, Escherichia coli, Proteus vulgaris,

Salmonella typhimurium and Pseudomonas aeruginosa. The hexane and

aqueous extracts were completely devoid of any activity.81

Antifungal

The acetone extract of the bark inhibited the condial germination of

Pyricularia oryzae and Colletotrichum falcatum at the concentration of 10.0

g/l and to a lesser extent at a concentration of 1.0g/l. It was toxic at the

concentration of 0.1 g/l 82. The ethyl acetate soluble fraction exhibited

fungitoxicity against Pyricularia oryazae comparable to that exhibited by

condensed type of tannins obtained from the bark.67

The bark and leaf decoction of the plant inhibited the

polygalacturonase enzyme activity of Alternaria tenuis indicating the

presence of tannins or phenolic compounds.83 The polyphenolic complex of

the bark at a concentration of 50 % showed maximum growth inhibition ( 56

%) as compared to controls against Fusarium oxysporum. The extract at 10

and 25 % dilution showed 24.0 % and 37.0 % inhibition, respectively in in

vitro studies.84



45

Combined Studies

In vitro antibacterial and antifungal activities of some metal arabates

(mercury, silver and copper) prepared from arabic acid of gum acacia showed

antimicrobial activity against Escherichia coli, Bacillus subtilis, Bac

anthracis, Bac pumilus, Salmonella typhosa, Staphylococcus aureus,

Micrococcus pyogenes, Proteus vulgaris and Aspergillus niger, A. flavus,

Trichophyton equingia, Fusarium oxysporum and Cryptococcus neoformans.

Both the activities were less as compared to the standard drugs penicillin

and salicylic acid respectively85. The fatty oil and unsaponifiable matter of

the seeds possessed antibacterial activity against Escherichia coli, Bacillus

anthracis, Bac subtilis, Corynebacterium pyogenes, Pseudomonas

aeruginosa and Staphylococcus aureus and antifungal activity against

Fusarium solani, F. moniliforme, Helminthosporium oryzae, H. turcicum,

Alternaria helianthi and Collectotrichum capsici. The fatty oil and the

unsaponifiable matter were less active as compared to penicillin and

streptomycin (antibacterial) and salicylic acid and resorcinol (antifungal) as

standards.86

Nematicidal

The aqueous leaf extract of the plant as also of A. Senegal showed

nematicidal activity against Meloidogyne incogenita as it inhibited its

hatching.87

Antiplaque Activity

The plant extract showed promising antiplaque activity in 12 male

students ranging from 19 to 24 years having healthy gingivae in a

comparative study made with eight new formulations of tooth powders.88



46

3.3. Balanites roxburghii linn. Fam:- Balanitaceae, Simaroubaceae


Sansk.: Angavriksha Hind.: Hingan, Hingoli. Kann.: Ingalore, Ingalkai Mar. :

Hinganbet Mal.:Nanjunta Guj.: Ingoriyo Tel.: Gara Ori.: Ingudihala, Beng.:

Hingon

Habit and Habitat

It is distributed in the drier parts of Pennisular India, western

Rajasthan and from South-East Punjab to West Bengal and Sikkim.89

Chemical studies

The fruit pulp contains five steroidal saponins, designated as

balanitism A, B, C, D and E. Balanitism A has been identified as diosgenin-

3-O-α-D-glucopyranosyl(1-3)-O-α-D-glucopyranosyl-(1�4)-O-L-ramnopyran-

oside. The diosgenin content of the fruits varies from 0.3 to 3.8 %.90

The leaves contain saponin of diosgenin, sigmasterol and free

diosgenin. Seeds are rich in protein, the amino acid composition of which is

reported. The oil from kernel contains linoleic acid, oleic, palmitic acid,

steric acid and balanitisins F and G. Deltoin, protodeltoin and a new

saponin, (25R and S)-spirost-5-en-3β-01; balanitos are present in stem bark,

balanitisin H and I are in stem-wood, 3-O-α-rhamnopyranosyl, (1�2)-β-D-

glucopyranosyl, (1�3)-β-D-glucopyranosyl, (1�4)-β-D-glucopyranoside are

reported in root-wood and six flavonoid glucosides have also been reported.91

Diosgenin (1.26%) are isolated from plant grow in Sudan. Diosgenin

content in leaves was lowest in May (0.05%) and highest in December

(0.47%). Two new saponins I & II characterized as 25(R) and 25(S) spirot-5-

en-3β-01-3-0- [α-L-rhamnopyranosyl (1�2)]-[β-D-glucopyranosyl (1�3)-β-D-

glucopyranosyl (1�4)]-β-D-glucopyranoside respectively are present in stem



47

bark in the ratio of 1.5:1. Deltonin and prodeltonin are also isoalted.

Quercetin-3-glucopyranosyde, quercetin-3-rutinoside, isorhamnetin-3-

glucoside and isorhamnetin-3,7-diglucoside are isolated from leaves and

branches whereas isorhamnetin-3-rutinoside and 3-rhamogalactoside are

isolated from leaves, branches and fruits. Compounds isolated from stem

bark exhibited insect antifeedant activity.

Dawidar and his co-workers compared diosgenin content of Balanites

roxburghii plant grow in Sudan as that of Egypt. They studied fleshy parts of

the fruits (epicarp and mesocarp) as a diosgenin source. Their results

revealed that the fleshy parts of the fruits of Balanites aegyptiaca grown in

Sudan contain an attractive % of diosgenin as a sole product nearly 1.267

while fruit from Egypt contains 0.3.92

Dawidar and Fayez found five diosgenin glucosides from leaves and

kernels of Balanites aegyptiaca including di, tri- and tetraglucoside. On

hydrolysis of the saponins they got 25D-spirosta-3,5-diene and 3β-chloro-

25D-spirost-5-ene as artifacts accompanying diosgenin. The unsaponifiable

fraction of the leaf fat contains two C29 sterols, one of which has an 8: 14

double bond as suggested by mass spectroscopy.93

Yamogenin was isolated as major sapogenin in these plant parts from

root, soft fruit wall (epicarp and mesocarp) and the stem bark. Infrared

spectrophotometric analysis showed 1% total steroidal sapogenin in the root,

0.7% in the stem bark and 1.2% in the soft fruit wall of Balanites

aegyptiaca Del, on moisture free basis. Elemental analysis of the sapogenin



48

and its derivatives as well as Mass, I.R. and NMR spectroscopy data show

that major sapogenin in this plant part is Yamogenin.94

Five new saponins named as Balanitisin A, B, C, D and E were

isolated from Balanites roxburghii (Planch) by Varshney and Co-workers95.

Balanitisin A contains 2 moles of glucose and 1 mole of rhamnose attached

at C-3 hydroxyl group of Diosgenin.The structure of Balanitisin A has been

establishedas diosgenin-3-O-α-D-glucopyranosyl(1�3)-O-α-D-glucopyranosyl

(1�4)-OH-L-rhamnopyranoside.

O

O

Me

O

O

H OH

CH2OH

O

O

OH

CH2OH

OHOH O

O

OH

CH2OH

OH

Me

Balanitisin A

Diosgenin from Balanites roxburghii leaves and kernels is also

reported96. The alcoholic extract of the crushed kernel of Balanites

roxburghii yielded an amorphous saponin which on acidic hydrolysis gave

mixture of three genins, out of which only the major one could be identified

as Diosgenin through mp and superimposible I.R. spectra.96



49

The isolation of the steroidal sapogenin, Diosgenin (25(R) spirost-5en-

3-β-ol], Yamogenin, β-sitosterol, Stigmasterol and β-sitosterol-3-O-β-D-

glucoside were recorded from the stem bark of Balanites roxburghii.95

The new saponin with insect antifeedant activity has been isolated

from the stem bark of Balanites roxburghii and characterized as (25(R)

spirost-5en-3-β-ol], 3-O-[α-L-rhamnopyranosyl (1�2)]-[β-D-glucopyranosyl

(1�3)- β-D-glucopyranosyl (1�4)]- β-D-glucopyranoside. In addition, two

other saponins were identified as the known saponins Deltonin and

Prideltonin.97

Brain and co-worker have performed rapid quantitative estimation of

25 α and 25 β-sapogenin individually and together in a crude plant extract.

The procedure is appropriate for the study of the variation in the C25 epimers

in morphological parts and with season, as well as for routine screening for

steroidal sapogenin of economic importance.

Five steroidal glycosides were isolated from the fresh caudex of Yucca

gloriosa together with YG-I and PS-I previsously obtained form flowers and

the structures of these glycoside were established. These are named as YS-

II,YS-III,YS-IV and YS-V. On acid hydrolysis five signals of steroidal

sapogenins was detected.98

Therapeutic Uses:-

Fruits: Unripe fruits are anthelmintic and cathartic, useful in curing

skin diseases and whooping cough when ripe; massage on chest with the

pulp mixed with goat-milk is known to be highly beneficial in the treatment

of pneumonia in children. The acidic fruit juice is used to remove stains on

clothes and for cleaning silk and cotton as detergent.



50

Seeds: Expectorant, efficacious in colic and whooping cough, oil

having antibacterial and anti-fungal properties is used for burns,

excoriations, freckles and other skin diseases.

Leave and Bark: Anthelmintic and purgative root emetic.99

Wood: The wood is used for making walking sticks and handles for

implements.100

PHARMACOLOGICAL STUDIES

The kernels yield bland, yellow oil (36-43%), which possesses anti-

bacterial and anti-fungal activity and is reported to be used in the treatment

of skin diseases, burns, excoriations and also freckles. It can be used for

production of soap.

The defatted seed meal is rich in protein (54.6%), having high content

of lysine. Other amino acids present are alanin 3.77 %, isodanine 4.12 %,

aspartic acid 9.21 %, Cysteine 0.56 %, glutamic acid 20.22 %, glycine 4.22

%, hystidine 1.4 %, leucine 7.9 %. The defatted seeds are source of

diosgenin. Also the fruit wall, which contains 1.5 % diosgenin on fresh weight

basis. The diosgenin content of the roots varies from 0.3 to 1.5%.101

Efficacy of daily oral feeding of crude alcohol extract from the

pericarp of unripe fruits of Balanites roxburghii at a dose of 500mg/kg for

45 days has been reported with respect to epididymal and testicular

histophysiology, metabolism and the fertilities potential of mice, following

withdrawal study. The testicular and cauda epididymal parameters revealed

altered physiology and metabolism resulting in a reduction in fertility of

extract fed mice.102



51

3.4. Achyranthus aspera Linn. Fam:- Amaranthaceae


Ben. Apang, Chirchiti; Eng.: Prickly-Chaff Flower, Rough Chaff tree; Guj.:

Angheda, safed Aghedo, Anghedo; Hind.: Chirchira, Chirchitta, Latjira,

Apamarg; Kan. Utranigida, Uttaranee; Mal.: Kadaladi; Mar.:Aghada; Ori.:

Apamaranga, Apamargo; Punj.: Chichra, Kutri; Tam.: chirukadaladi,

Naayurivi, Tel.: Apamargamu, Uttareeni, Uttaraene, Uttareni

Habit and Habitat

An erect or procumbent, annual or perennial herb, often with a woody base

commonly found as a weed on waysides and wet places throughout India up

to an altitude of 2100m.

Ayurvedic Description

Sankrit name : Apamarga

Synonyms : Sikhari, Adhahsalya, Mayuraka, Pratyakpuspa,

Kharamanjari

Properties : Rasa: katu, tikta; Guna: laghu, tiksna, sara;

Virya: Usna; Vipaka : Katu

Action : Dipana, pacana, kaphamedahara, vamanopaga,

vranasodhaka, raktasamgrahi, visaghna.

Therapeutic uses : Krmi, arsa, kandu, asmari

Therapeutic Uses

The dried plant is used in sula (colic), udararoga (diseases of the

abdomen), apaci (lymphadenitis-cervical), arsa (haemorrhoids), kandu

(itching), medoroga (obesity). The dried root of the plant is used in chardi

(vomiting), adhmana (tympanitis), kandu (itching), sula (colic), apaci



52

(lymphadenitis), granthi ( tumor), bhagandara (fishula-in-ano), hrdaroga

(diseases of heart), jwara (pyrexia), switra (leucoderma), vadhirya (deafness),

udararoga (diseases of the abdomen), yakratroga ( disorders of the liver),

dantaroga ( diseases of tooth), raktavikara (blood disorders).103

Properties and Uses Ascribed

The plant is reported to be pungent, astringent, pectoral and diuretic.

It is used as an emmenagogue in piles and skin eruptions. The decoction of

the plant is useful in pneumonia and renal dropsy while the juice of the

plant is used in opthalmia and dysentery. The leaves are used as a cure for

gonorrhoea and excessive perspiration. Their extract is used for leprosy and

the heated sap for tetanus. The root is astringent, the paste is applied to

clear opacity of cornea and to wounds as a haemostatic. The root is also

reported to be useful in cancer. A decoction of the root is used for stomach

troubles and an aqueous extract for stones in the bladder. The flowers,

ground and mixed with curd and sugar are given for menorrhagia. The flower

tops are stated to be employed for the treatment of rabies. Powdered seeds

are soaked in butter milk and given for biliousness. The seeds are said to be

emetic and used in hydrophobia.103

Ethnobotanical Studies

The plant is used in dropsy, piles, skin eruptions, colic, as a diuretic,

astringent and purgative104 in heart diseases, ascites, uterine-bledding105 as

diuretic, in piles, dysentery, in fractured bones, intestinal parasites,

whooping cough, respiratory troubles, 106 for ashma, 107,108 in abdominal

disorders and as antitumour agent, 109 in renal disorders110 , mild and

malarial fever111 , as a laxative112 and in leucoderma113 . The inflorescence is



53

used in cough. The stem is used in pyrrhoea and scorpion bite while the

twigs are used in toothache, in arthritis114. Fruits are used in hydrophobia.

The seeds are employed in bleeding piles, as an emetic, purgative, cathartic,

in gonorrhoea, for insect bites, snake bite and in hydrophobia115,for scorpion

sting, in skin diseases, cough including whooping cough, as an

antiasthmatic, in diarrhoea, as an astringent, diuretic and laxative. These

are also used in renal dropsy, bronchitis, as antileprotic and in gastric

troubles. The leaves are used in wounds, injuries, as an antidote to insect

bite and scorpion sting, in cough, toothache, excessive bleeding in

menstruction, in urination, as eye drops, in breast ulcers, backache, cuts,

bolis, blisters; inclammed ears and eyes, leucoderma, amoebiasis, dysentery,

diarrhoea, dog bite, as antimalarial, for aprains, as antipyretic and

antirickettsial, in skin diseases and liver enlargement, in stomachache and

in typhoid. The root is used in body pain including waist and ribs pain,

labour pain, as antipyretic, antirickettsial, as a tonic, in pyrrhoea, as a

contraceptive, for ante-partum treatment, whooping cough, tonsillitis, in

diarrhoea, menstrual disorder; burn healing, for snake bite and scorpion

sting, as an astringent, chronic fever, renal troubles, sensitive gums,

haemorrhage, leucorrhoea, in cough, hydrophobia, as antiasthmatic, as a

galactagogue, for dental problems, spermatorrhoea, cataract, stomach ache,

in constipation, piles, as haemostatic, in malarial fever, diarrhoea, vomiting;

as diuretic, diaphoretic, antisyphilitic, in toothache, for dandruff, hair

problems, acne/pimples, in leprosy and as an antiseptic after delivery.103



54

CHEMICAL STUDIES

The plants were reported to yield a water-soluble base and a

chloroform-soluble base. The former was earlier designated as

achyranthine116 and was characterized as a betaine derivative of N-

methylpyrrolidine-3 carboxylic acid. Later studies showed that the water-

soluble base was betaine and not achyranthine117. The chloroform-soluble

basic fraction was shown to be a mixture of two uncharacterized alkaloidal

entities.118

The ethanolic extract of the plant parts contained alkaloids and

saponins while flavonoids and tannins were found absent.119

The shoot yielded a new aliphatic dihydroxyketone, characterized as

36, 47-dihydroxyhenpentacontan-4-one together with tricontanol; an

essential oil, a new long chain alcohol characterized as 17-

pentatriacontanol, four new compounds characterized as 27-

cyclohexylheptacocsan 7-ol, 16-hydroxy-26-methylheptacosan-2-one, 4-

methylheptatriacont-1-en-10-ol and tetracontanol-2.120

Various parts of the plant viz. seeds, stem, leaves and root were

reported to contain ecdysterone. The chloroform extract of the stem led to the

isolation of pentatriacontane, 6-pentatriacontanone, hexatriacontane and

triacontane. The inflorescence was reported to contain flavonoids and

alkaloids.121

The food value of the seeds in terms of its protein quality was studied.

The composition of the seeds showed close similarity to Bengal gram with a

protein content of 24.8% and calorific value of 3.92/g. The hydrolysate

contained the usual amino acids. The values obtained for ten essential amino



55

acids and cystin showed that the seed protein compared favourable with

Bengal gram in its leucine, isoleucine, phenylalanine and valine content,

while its tryptophan and sulphur amino acid (methionine and cystine)

content were higher than most of the pulses. It was, however, deficient in

arginine, lysine and theronine as compared to the whole egg protein.122

The defatted seeds were reported to yield a saponin in an yield of 2 %

which was identified as oleanolic acid, oligosaccharide. The sugar moiety of

the saponins was composed of glucose, galactose, xylose and rhamnose123.

Khastgir and associates isolated a crude sapogenin fraction from the seeds,

which yielded oleanolic acid. Later, investigations led to the isolation of two

oleanolic acid based saponins, saponin A and saponin B which were

characterized as α-L rhamnopyranosyl (1�4) – β-D-glucopyranosyl (1�4)- β-

D-glucuronopyranosyl (1�3)- oleanolic acid β-D-galactopyranosyl (1�28)

ester of saponin A, respectively124. In another study, the total saponins were

hydrolysed with acid and the genin was identified as oleanolic acid125. A

rapid procedure for the separation of triterpenoid saponins based on

partition chromatography from the plant has been described126. The seeds

contained hentriacontne, 10-octacosanone, 10-triacosanone and 4-

tritriacontanone.127

The fatty oil constituents of the seed oil comprised of lauric, myristic,

palmitic, stearic, arachidic, behenic, oleic and linoleic acids.128

The unripe fruits yielded two new saponins (C and D ) which were

identified as β-D-glucopyranosyl ester of α-L-rhamnopyranosyl (1�4) -β-D-

glucuronopyranosyl (1�3)-oleanolic acid and β-D-glocopyranosyl ester of α-



56

L-rhamnopyranosyl (1�4) –β-D- glycopyranosyl(1�4) –β-D-

glucuronopyranosyl (1�3) oleanolic acid respectively.129

The chemical constituents of the root varied in different preliminary

studies carried out. The root was found to contain oleanolic acid as the

aglycone from the saponin fraction130. Both root and shoot of the plant were

found to contain saponins and alkaloids but no flavonoids131. In another

study the root of the plant was found to contain alkaloids but indicated

absence of saponins and tannins132. In yet another preliminary chemical

study the root was reported to contain alkaloids, flavonoids, saponinis,

steroids and terpenoids. Glycosides were found to be absent133. Isolation of β-

sitosterol was also reported from the root.134

PHARMACOLOGICAL AND BIOLOGICAL STUDIES

General pharmacological studies of the plant did not elicit any exciting

activity. However, the antifertility activity needs to be looked into. The plant

seems to lower lipids.

Antifertility

The alkaloidal fraction obtained from the alcoholic extract of the root

bark inhibited the response of oxytocin in isolated rat uterus. This fraction

did not inhibit the responses to serotonin and acetylcholine in rat uterus and

to histamine in guinea pig uterus.135

The crude benzene extract of the stem was found to have potent

abortifacient effect in mice136. In an attempt to locate the active principle,

various chromatographic fractions were tested for antifertility activity in

female mice. The maximal activity was found to be located in the fraction

eluted with 50 percent benzene in petroleum ether.137



57

The benzene eluate of the benzene extract of the plant showed 100 %

abortifacient activity in rabbit at the single dose of 50 mg/kg b.w. In a study

on the possible mode of action, the extract was found neigher estrogenic nor

antiestrogenic or androgenic in mice. No teratogenicity was observed138. The

benzene extract of the plant (excluding root) also revealed 66.6 %

antiimplantation activity in female albino rats139. The ethanolic extract of the

plant (excluding root) at a dose of 100-200 mg/kg b.w. administered orally

revealed 60 % antifertility activity on early pregnancy in rats. Further, the

plant also showed potent activity at secondary testing level.140

The methanolic extract of the root revealed 60 % antiimplantation

activity in rats while the acetone extract of the root prevented implantation in

50% of rats.141

The n-butanol fraction of the aerial parts prevented pregnancy in

adult female rats when administratered orally daily dose of 75 mg/kg or

more on 1-5day post coitum, but was ineffective in hamsters up to 300

mg/kg dose. Antifertility activity was not observed in the aqueous fraction in

either rats or hamsters. In ovariectomized immature female rats, the extract

exhibited potent estrogenic activity at the dose of 75 mg/kg. It induced even

at a does of 3.75 mg/kg.142

Antitumour

In an antitumour evaluation of Acryanthus aspera, a sterol glycoside

(mp 280-85oC) isolated from the plant did not show any antitumour activity

against P-388 tumour in mice.143



58

Liver and Lipids

The alcoholic extract of the plant at 100 mg/kg dose lowered total

serum cholesterol (TC) and phospholipids (PL), triglyceride (TG) and total

lipids (TL) levels by 60, 51, 33 and 53 %, respectively in triton-induced

hyperlipidemic rats. The chronic administration of the extract at the same

doses to normal rats for 30days, lowered serum TC,PL, TG and TL by 56, 62,

68 and 67 % respectively followed by significant reduction in the levels of

hepatic lipids. The faecal excretion of cholic acid and deoxycholic acid

increased by 24 and 40 % respectively under the action of the drug. The

possible mechanism of action of cholesterol lowering activity of the plant

might be due to rapid excertion of bile acids causing low absorption of

cholesterol.144

Antiinflammatory

The water soluble alkaloid achyranthine was screened for its anti-

inflammatory and antiarthritic activity against carrageenin-induced foot

oedema, granuloma pouch, formalin-induced arthritis and adjuvant arthritis

in rats. It showed significant anti-inflammatory activity in all the four models

employed but was less active than phenylbutazone and betamethasone.

Further, achyranthine significantly reduced the weight of adrenal gland,

thymus, spleen and raised the adrenal ascorbic acid and cholesterol

contents. The effects were qualitatively similar to betamethasone. All the

three drugs tested reduced food intake but had no significant effect on

urinary and faecal output and on mortality rate. Incidence of gastric ulcers

was maximum with betamethasone and minimum with achyranthine.145



59

Diuretic

Earlier studies by Bhide et al on potassium diuretics indicated that

the diuretic activity of the ash of the plant may be due to high potassium (44

%) content.146

Effect of a saponin isolated from the seeds on urine output in albino

rats has been investigated in comparison to mersalyl and acetazolamide. The

saponin in 10-20 mg/kg i.m. doses caused significant increase in urine

output after 2, 6 and 24h as compared to untreated control rats. The diuretic

effect was comparable to that observed with 3 mg/kg dose of mersalyl. The

optimum dose of the saponin was 10 mg/kg. After oral administration of the

saponin (5-10 mg/kg) in rats a significant increase in urine output was

observed which was comparable to that of 10 mg/kg oral dose of

acetazolamide. The diluretic effect of the saponin like acetazolamide was

associated with an increase in the excretion of sodium and potassium in the

urine.147


In a preliminary study the aqueous and alcoholic extracts of the root

caused a sharp and transient fall in blood pressure without any significant

action on the respiration of anaesthetized dogs. In higher doses there was

slight respiratory depression. Atropine sulphate blocked the hypotensive

effect of the extracts. On frog’s heart the extracts had a temporary negative

inotropic and chronotropic effects. The extracts produced spasm of isolated

rabbits ileum increased the tone and amplitude of contractions in gravid and

non-gravid uteri of albino rats, guinea pigs and rabbits. Oral administration

of the drug significantly increased the urinary output in rabbits.148



60

The total chloroform soluble basic fraction ( alkaloidal residue)

obtained from the plant raised the blood pressure of anaesthetized dog

caused initial transitory stimulation of respiratioin and increased the

amplitude of cardiac contractions of isolated guinea pigs heart. It showed

spasmolytic action against various spamogens on intestine and uterine

muscles of guinea pigs and a slight antidiuretic action in rats. No specific

CNS effects were observed in mice. The fraction did not possess analgesic

activity in rats. The water soluble alkaloid, achyranthine isolated from the

plant was found to lower blood pressure, depress the heart, dilate the blood

vessels and increase the rate and amplitude of respiration in anaesthetized

dogs. It showed spasmogenic effect on frog’s rectus muscle and diuretic as

well as purgative active in the albino rats. No effect was observed on isolated

rabbit, guinea pig and rat ileum and on CNS. The drug exerted a slight

antipyretic effect.149,150

The mixture of saponins isolated from the seeds caused a significant

increase in force of contractions of the isolated heart of frog, guinea pig and

rabbit. The stimulant effect of the lower dose (1 to 50 µg) of the saponins was

blocked by pronethalol and partly by mepyramine. The effect of higher doses

was not blocked by pronethalol. The saponin increased the tone of the

hypodynamic heart and also the force of contraction of failing papillary

muscle. The effect was quicker in onset and shorter in duration in

comparison to that exerted by digoxin151. The effect of saponin on the

phosphorylase activity of the perfused rat heart has been investigated and

compared with that of adrenaline. The saponin has been found to stimulate



61

the phosphorylase activity of the heart and its effect was comparable to that

of adrenaline.152

The ethanolic extracts of the plant153 and leaves154 were screened for

preliminary biological activities. The former extract showed hypoglyceamic

activity in rat. It was devoid of antibacterial, antifungal, antiprotozoal,

anthelmintic, antiviral and anticancer activities and effects on isolated

guinea pig ileum, respiration, CVS and CNS in experimental animals. The

MTD of the extract was found to be 1000 mg/kg b.w. orally in mice. The leaf

extract was found to be devoid of antiprotozoal and antiviral activities and

effects on respiration, preganglionically stimulated nictitating membrane,

CVS and CNS in experimental studies. The LD50 of the latter extract was

greater than 100 mg/kg i.e. in mice.154

Toxicity

An alkaloid isolated from the plant was tested for its acute, subacute

and chronic toxicity in rats. During acute toxicity test, there was a slight

increase in sedation and slight loss in wrighting region at 6.0 mg/kg. dose

level which became prominent at 7.0 mg/kg dose level. At higher doses,

significant deplection in wrighting region, depression in respiration,

remarkable increase in sedation and diarrhoea was observed. Subacute

toxicity test revealed (5.0 and 6.0 mg/kg.) a significant increase in sedation

and hypnosis, depletion in respiration and loss of brighting reflexes. At 6.0

mg/kg dose, it also caused remarkable increase in salivation and diarrhoea.

Chronic toxicity showed (3.0 mg/kg.) an increase in sedation, hypnosis,

salivation and diarrhoea. These was a significant depression of respiration

and loss of body weight.155



62

Antimicrobial

The aqueous solution of the base achyranthine as well as the entire

plant showed antibacterial activity against Staphylococcus aureus,

Streptococcus haemolyticus and Bacillus typhosus156 while the alcoholic and

the aqueous extract of the leaves showed antibacterial activity against

Staphylococcus aureus and Escherichia coli.157

The seeds growing on cattle dunk revealed antibacterial activity

against bacterial strains of Bacillus subtilis, Pseudomonas cichorii and

Salmonella typhimurium158. In another study, the 80 % ethanolic extract of

the leaves and stem of the plant inhibited Bacillus subtilis and

Straphylococcus aureus bacterial strains at a concentration of 25 mg/ml.159

The aqueous leaf extract in in vitro studies showed antibacterial

activity against Proteus vulgaris at concentration of 5000 ppm. The extract

inactive against Klebsiella aerogenes, Pseudomonas aeruginosa and

Escherichia coli160. The aqueous residues of another sample of the plant

levels were found devoid of any activity against Alkaligenes viscolactis,

Aeromonas hydrophilla, Cytophaga sp., Vibrio parahaemolytica, Vib damsela,

Bacillus cereus and Streptococcus pyogenes in addition to Escherichia coli,

Klebsiella aerogenes, Pseudomonas aeruginosa161. In another study the

extract of the leaves was found to be active against the isolated bacteria

Escherichia coli, Staphylococcus citri and aerobic spore formers from soft

drinks.162

The deproteinised leaf extract of the plant affected the spore

germination of the plant fungi viz., Alternaria brassicicola, Helminthosporium



63

apattarnae, Pestalotia sp., Penicillum purpurogenum, Aspergillus niger,

Trichotheciym sp., Neurospora sp., Fusarium sp., Trichoderma sp. and

Rhizopus sp.163. The aqueous extract of the leaves in treated bananas showed

poor activity against the infested fungi Botryodiploidia theobromae, Fusarium

oxysporum, Helminthosporium speciferum, curvalaria lunata, Aspergillus

flavus and Trichothecium roseum causing banana rot.164

The extract of the leaves revealed colatile fungitoxicity against the

damping off fungi Pythium aphanidermatum, P. debaryanum and Rhizoctonia

solani. The extract was neigher inhibitory nor stimulatory against the

conidial germination of Drechslera oryazae.165,166

The essential oil isolated from the shoot was reported to have

antifungal activity against Aspergillus carneus at various concentrations. The

oil showed 85-100 per cent inhibition of the mycelial growth at concentration

ranging between 1000-5000 ppm.167

In a comparative study of herbal agents used for fumigation in relation

to formalin, the plant reduced the microbial colony counts in air samples

considerably.168

Anthelmintic

Alcoholic extract of the plant did not show any effect on human

Ascaris lumbricoides in vitro169. The leaf extract at a dilution of 1:5 showed

100 % mortality against Meloidogyne Javanica.170

Insecticidal

The crude extract of the plant did not show juvenoid activity against

nymphus of Dysdercus cingulatus.171



64

CLINICAL STUDIES

The plant was subjected to wide clinical evaluation with special

reference to its use in leprosy, bronchial asthma and fistula-in-ano. Diuretic

activity could not be confirmed.

Leprosy

The effect of oral decoction of Acryanthus aspera in the treatment of

leprosy was studied (uncontrolled) in 19 patients who were found to have

positive stain smears at the S.S. Hospital, Varanasi. Fourteen patients were

in stage of reaction and rest of them had active lesions but none of them was

in quiescent stage. The study revealed encouraging results in both lepra

reaction as well as the quiescent stage of lepromatous leprosy. Skin lesions

and ulcers had tendency to subside quickly with the treatment.The

bacteriological index had also improved.172

In an attempt to get additional data on the efficacy of the decoction of

Acryanthus aspera, a study was undertaken for its role in the management of

reactions in leprosy. An open clinical trial was conducted on twelve cases

selected from the leprosy clinic of S.S. Hospital, Banaras Hindu University,

Varanasi. Out of the twelve cases, six were of acute and subacute type of

lepra reaction, three of acute and subacute type of border line reaction, and

three of subacute type of tuberculoid reaction. It was observed that the

decoction was useful in the treatment of reactions in leprosy particularly in

subacute and mild type. When administered in conjunction with the

antileprosy drug diaminodiphenylsulphone (DDS), it was found that the

chances of reaction becames less and rate of improvement was faster. No



65

toxic manifestation which could be attributed to Acryanthus aspera was

noted during the trial.173

In another study, at S.S. Hospital, Varanasi, thirty six patients of

advanced, infiltrative and nodular type of lepromatous leprosy, who had no

previous treatment (as revealed by their histories), were selected for the trial.

The patients were randomly divided into three groups and administered (a)

the decoction of the plant in group 1; (b) DDS in group 2 and (c) the

decoction of the plant along with DDS in group 3. The decoction was

administered orally in a dose of 1.0 oz twice daily. Clinical and bacterial

assessments were carried out every three months and finally after one year.

During the whole treatment period a statistical change was noted in the

clinical condition, bacterial status and general health. All the patients,

irrespective of their grouping, showed clinical improvement of varying

degrees. It was observed that those patients who received DDS alone could

make a better clinical progress compared with those who received the

decoction of ‘apamarga’ alone, whereas the patients who received the

different drugs separately. The patients who received only the decoction

showed no improvement in their bacterial index. In fact most of them showed

slight deterioration. The patients on DDS alone and those on a combined

therapy of the decoction and DDS showed definite improvement in their

bacterial index, the latter group showed a greater improvement. An

improvement in general health was observed in all the patients receiving the

decoction. Those patients who were on DDS alone did not show any

improvement in general health. ‘Apamarga’ did not produce any toxic

manifestation in any patient and was well tolerated. The study revealed that



66

decoction of the plant had clinical effectiveness in the therapy of leprosy,

prevented the antileprosy action of DDS and reactions in leprosy.174

Fistula-in-ano

There have been a number of studies on the use of Kshaarasootra (a

medicated thread prepared by coating the latex of Euphorbia neriifolia,

alkaline powder of Acryanthus aspera and Curcuma longa in the treatment of

fistula-in-ano. The studies revealed that the long term use of Kshaarasootra

was quite effective in treatment of various fistulous tracks. The

standardization of Kshaarasootra the methods of identification and assay of

the individual constituents were studied.175, 176,177

Kshaarasootra has also been found to given encouraging results in five

patients of chronic non healing milk-fistula ‘stannadi-varna’ with additional

local application of ‘jatyaditaila’ and oral administration of ‘shigru guggulu’

(two tablets t.i.d) during the course of treatment.178

Diuretic

A clinical trial was undertaken at the Government Ayurvedic College

Hospital, Gwalior, involving fifteen cases of general anasarca (shoth) to

evaluate the diuretic effect of Acryanthus aspera. The decoction of plant, its

seed or saponin did not possess a significant diuretic property (immediate or

late or late diluresis) in cases of general anasarca.179

Bronchial Asthma

A pilot study was carried out at the Central Research Institute for

Siddha in Madras on fifteen cases of bronchial asthma. The oil obtained from

the root soaked in cows urine was smeared on betal leaf and administered

thrice a day to these patients. In most of the cases symptoms like wheezing,



67

gasping, dyspnoea, sneezing and cough disappeared. A fall in the total WBC

and eosinophil counts and ESR was observed.180

TOXICOLOGICAL STUDIES

The allergenic pollens of the plant are common in the area of

Pondicherry. The period of anthesis has been found to be between November

and December.181

3.5. Nyctanthes arbor-tristis Linn. Fam: - Oleaceae

Habit and Habitat

Drug consists of the dried leaf of Nyctanthes arbor-tristis Linn.Fam.

Oleaceae. A small tree occurring wild in the outer Himalayan ranges from

Kashmir to Nepal and almost throughout India up to 1000m. IT is also

cultivated in gardens and elsewhere.


Beng. -Sephalika, Seoli Eng.-Coral Jasmine Guj.-Jayaparvati Hind.-

Harsinghar, Seoli, Siharu Kan.-Harsing, Parijata Mal.-Pavizhamalli,

parijatakom Mar.-Khurasli, Parijatak Ori.-Godokodiko, Gunjoseyoli,

Singaroharo Tam.-Manjhapu, Pavazhamalligai, Pavalamallikai Tel.-

Parijatamu, Pagadamalli

Chemical Studies:-

Nicotiflorin182,183, nyctanthic acid182,184 are major constituents besides

this others are Astragalin182, friedelin, mannitol, Beta-sitosterol, lupeol,

oleanolic acid182, arborside A,B,C185, nyctoside186,

desrhamnosylverbascoside186,187, benzoic acid, hentriacontane, Beta- amyrin,

D- mannitol, glucose, fructose188, 6,7-di-O-benzoylnyctanthoside, 6-O-trans-



68

cinnamoyl-6B-hydroxyloganin, 7-O-trans-cinnamoyl-6 B-hydroxyloganin189

are also found.

Pharmacological studies

The extract of the leaves showed tranquillizing, hypothermic and

purgative effects190. The extract exhibited significant antipyretic, analgesic,

and ulcerogenic activities191.The alcoholic extract produced anti-

inflammatory activity192, 193 and blocked the stimulatory response of

acetylcholine and histamine in isolated rabbit ileum192. Oral administration

of the extract produced depletion of tumour necrosis factoralpha194, reduced

plasma interferon-gamma194 and showed immunostimulant property195. The

leaf extract showed antileishmanial activity in hamsters196. The alcoholic

extract possessed antimalarial197, antitrypanosomal198 and antiamoebic

activities. 199

Major Therapeutic Claims

Sciatica200, 201

Safety Aspects

Oral administration of the leaf extract for fix consecutive days induced

gastric ulcers in rats191. The extract also produced histological changes in

kidney, thymus, lymph node and lachrymal gland in rats.202

Dosage

Juice of leaves: 10 to 20 ml.201



69

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1993, 163-164. 165) Pandey, V.N.; and Dubey, N.K.; Biol Plant, 34, 1992, 143-147. 166) Ganesan, T.; and Krishnaraju, J.; Adv Plant Sci, 8, 1995, 194-196. 167) Misra, T.N.; Singh, R.S., Pandey, H.S., Prasad, C. and Singh, B.P.;

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1965, 41-43. 184) Dictionary of Organic Compounds, Vol. IV. London: Eyre and

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185) Srivastava, V.; Rathore A.; Ali, S.M.; Tandon, J.S.; J Nat Prod, 53, 1990,

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