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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
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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
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Department of Chemical Technology, Dr.Babasaheb Ambedkar Marathwada University, Aurangabad
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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,
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PLATE NO.1
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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
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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
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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
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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
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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
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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
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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
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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
Regional and Other Names
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.
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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
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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
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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
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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
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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 %).
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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
General Pharmacology
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
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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
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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
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3.3. Balanites roxburghii linn. Fam:- Balanitaceae, Simaroubaceae
Regional and Other Names
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
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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
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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
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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.
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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
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3.4. Achyranthus aspera Linn. Fam:- Amaranthaceae
Regional and Other Names
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
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(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
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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
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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
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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-
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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
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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
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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
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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
General Pharmacology
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
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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
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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
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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
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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
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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
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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
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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,
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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.
Regional and Other Names
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-
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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
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116) Basu, N.K.; J Proc Inst Chem, 29, 1957, 73-76. 117) Kapoor, V.K.; and Singh, H.; Indian J Chem 4, 1966, 461. 118) Kapoor, Y.K.; and Singh, H.; Indian J Pharm, 29, 1967, 285-288.
119) Kumar, S.; Singh, J.P.; and Kumar, S.; J Econ Bot Phytochem, 1, 1990, 13-16.
120) Misra, T.N.; Singh, R.S.; Pandey, H.S.; Prasad, C.; and Singh, S.; Indian
J Chem, 35B, 1996, 637-639. 121) Prasad, S.; and Bhattacharya, I.C.; J Sci Ind Res, 20C, 1961, 246-251. 122) Satyanarayana, M.S.; Susheela, B.A.; Rao, N.A.N.; and Vijayaraghavan,
P.K.; J Food Sci Technol, 1, 1964, 26-27. 123) Gopalachari, and Dhar, M.L., ; J Sci Ind Res, 11B, 1952,209. 124) Hariharan, V.; and Rangaswamy, S.; Phytochemistry, 9, 1970, 409-414. 125) Batta, A.K.; and Rangaswami, S.; Phytochemistry, 12, 1973, 214-216. 126) Sarkar, B.; and Rastogi, R.P.; J Sci Ind Res, 19B, 1960, 106-107. 127) Ali, M.; Oriental J Chem, 9, 1993, 84-85. 128) Daulatabad, C.D.; and Ankalgi, R.E.; Fette Seifen Anstrichm, 87, 1985,
196-197 (Cherm Abstr 1985 lO3, 34909p). 129) Seshadri, V.; Batta, A.K.; and Rangaswami, S.; Indian J Chem, 20B,
1981, 773-775. 130) Khastgir, H.N.; Sengupta, S.K.; and Sengupta, P.; J Indian.Chem Soc,
35, 1958, 693-694. 131) Sinha, S.K.P.; and Dogra, J.V.V.; Int J Crude Drug Res, 23, 1985, 77-
86. 132) Joshi, M.C.; and Sabnis, S.D.; Bull Med Ethnobot Res, 10, 1989, 61-82. 133) Agrawal, R.G.; Pant, P.; (nee Samwal), Tewari, L.C.; Singh, J.; Pandey,
M.J.; and Tiwary, D.N.; Bull Med Ethnobot Res, 10, 1989, 176-186. 134) Misra, T.N.; Singh, R.S.; Pandey, H.S.; Prasad, C.; and Singh, B.P.;
Phytochemistry 33, 1993, 221-223. 135) Gupta, S.S.; and Khanijo, I.; Indian J Physiol Pharmacol, 14, 1970, 63. 136) Pakrashi, A.; Mookerji, N.; and Basak, B.; J Reprod Fert, 43, 1975a,
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127-128. 137) Pakrashi, A.; Basak, B. and Mookerji, N.; Indian J Med Res, 63, 1975b,
378-381. 138) Pakrashi, A.; and Bhattacharya, N.; (nee Mookerji); Indian J Exp Biol,
15, 1977, 856-858. 139) Mathur, R.; Chauhan, S.; Saxena, V; Shukla, S.; and Prakash, A.O.; J
Jiwaji Univ (Sci Technol.Med), 9, 1983, 37-46. 140) Prakash, A.O.; Shukla, S.; and Mathur, R.; Camp Physiol Ecol, 12,
1987, 157-171. 141) Prakash, A.O.; Int J Crude Drug Res, 24, 1986, 19-24. 142) Wadhwa, V.; Singh, M.M.; Gupta, D.N.; Singh, C. and Kamboj, V.P.;
Planta Med, No 5, 1986, 231-232. 143) Indap, M.A.; Ambaye, R.Y.; and Bhat, M.L.; Indian Drugs, 23, 1986, 477. 144) Khanna, A.K.; Chander, R.; Singh, C.; Srivastava, A.K.; and Kapoor, N.K.;
Indian J Exp Biol, 30, 1992, 128-130. 145) Neogi, N.C.; Rathor, R.S.; Shreshtha, A.D.; and Banerjee, B.K.; Indian J
Pharmacol, 1, 1969, 37-47. 146) Bhide, N.K.; Altekar, W.W.; Trivedi, J.C.; and Sheth, U.K.; J Postgrad
Med, 4, 1958, 21-27. 147) Gupta, S.S.; Verma, S.C.L.; Ram, A.K.; and Tripathi, R.M.; Indian J
Pharmacol, 4, 1972b, 208-214. 148) Gambhir, S.S.; Sanyal, A.K.; and Chowdhury, N.K.; Indian J Physiol
Pharmacol, 9, 1965, 185-188. 149) Basu, N.K.; Singh, H.K.; and Aggarwal, O.P.; J proc Inst Chem, 29,
1957a, 55-58. 150) Neogi, N.C.; Garg, R.D.; and Rathor, R.S.; Indian J Pharm, 32, 1970, 43-
46. 151) Gupta, S.S.; Bhagwat, A.W.; and Ram, A.K.; Indian J Med Res, 60, I
972a, 462-471. 152) Ram, A.K.; Bhagwat, A.W.; and Gupta, S.S.; Indian J Physiol
Pharmacol, 15, 1971, 107-110. 153) Dhar, M.L.; Dhar, M.M.; Dhawan, B.N.; Mehrotra, B.N.; and Ray, C.;
Indian J Exp Biol, 6, 1968, 232-247.
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154) Aswal, B.S.; Goel, A.K.; Kulshrestha, D.K.; Mehrotra, B.N.; and Patnaik,
G.K.; Indian J Exp Biol, 34, 1996, 444- 467. 155) Mali, A.; Maheshwari, H.S.; and Santani, D.O.; J Indian Acad Forensic
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157) George, M.; Venkataraman, P.R.; and Pandalai, K.M.; J Sci Ind Res, 6B,
1947, 42-46. 158) Sushil Kumar, Bagchi, G.D.; and Darokar, M.P.; Int J Pharmacog, 35,
1997, 179-184. 159) Basu, N.K.; Neogi, N.C. and Srivastava, V.P.; J Proc Inst Chem, 29,
1957b, 161- 165. 159) Valsaraj, R.; Pushpangadan, P.; Smitt, U.W.; Adsersen, A.; and Nyman,
U.; J Ethnopharmacol, 58, 1997, 75-83. 160) Perumal Samy, R.; Ignacimuthu, S.; and Sen, A.; J Ethnopharmacol,
62, 1998, 173-182. 161) Perumal Samy, R.; Ignacimuthu, S.; and Patric Raja, D.; J
Ethnopharmacol, 66, 1999, 235-240. 162) Meera, P.; Anita Dora, P.; and Karunyal Samuel, J.; Geobios, 26, 1999,
17-23. 163) Mukadam, D.S.; Balkhande, L.D.; and Umalkar, G.V.; Indian J
Microbiol, 16, 1976, 78-79. 164) Singh, H.N.P.; Prasad, M.M.; and Sinha, K.K.; J Indian Bot Sac, 72,
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.;
Phytochemistry 31, 1992, 1811-1812. 168) Bisht, L.S.B.; Brindavanam, N.B.; and Kimothi, G. P.; Ancient Sci Life,
8, 1988, 125-132. 169) Kaleysa Raj, R.; Indian J Physiol Pharmacol, 19, 1975, 47-49. 170) Nandal, S.N.; and Bhatti, D.S.; Indian J Nematol, 13, 1983, 123-127. 171) Srivastava, V.S.; Jaiswal, A.K.; and Abidi, R.; Curr Sci, 54, 1985, 576-
578.
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172) Tripathi, S.N.; Chaturvedi, G.N.; and Dube, G.P.; J Med Sci (BHU), 4,
1963, 103-1 12. 173) Ojha, D.; Tripathi, S.N.; and Singh, G.; Lep Rev, 37, 1966, 115-120.
174) Ojha, D.; and Singh, G.; Lep Rev, 39, 1968, 23-30. 175) Dwivedi, A.K.; Chaudhury, M.; Kulshreshtha, D.K.; and Sarin, J.P.S.; J
Res Ayur Siddha, 12(1-2), 1991, 85-92. 176) Sharma, K.P., Kushwah, H.K. and Patil, J.Y.; Sachitra Ayurved 47,
1994, 685-688. 177) Sharma, A.; Mehta, M.; Handa, S.S.; Dwivedi, V.K.; Prakash, P.; Singh,
S.; and Sarin, J.P.; Ksharasutra Therapy in Fistula-in-ano and Other Anorectal Disorders.” Eds. Sharma, S. K., Sharma, K.R. and Singh, K. Rashtriya Ayurved Vidyapeeth, 1995, 184-203.
178) Singh, S.K.; Singh, S.; and Sijoria, K.K.; J Res Ayur Siddha, 15, 1994,
150-154. 179) Shankar, A.; Parsai, M.R.; Naqvi, S.M.A.; and Jain, J.P.; J Res Ayur
Siddha, 1, 1980, 514-527. 180) Suresh, A.; Anandan, T.; Sivanandam, G.; and Veluchamy, G.; J Res
Ayur Siddha, 6, 1985, 171-176. 181) Saha, J.C.; and Kalyanasundaram, S.; Indian J Med Res, 50, 1984,
881-888. 182) Anjaneyulu, A.S.R.; Murty, Y.L.N.; Row, L.R.; J Indian Chem Soc, 58,
1981, 817-818. 183) Singh, S.P.; Bhattacharji, S.; Sen, A.B.; Bull Natl Inst Sci India, 31,
1965, 41-43. 184) Dictionary of Organic Compounds, Vol. IV. London: Eyre and
Spottiswoode Publishers Ltd., E & F. N Span Ltd. New York: Oxford University Press.
185) Srivastava, V.; Rathore A.; Ali, S.M.; Tandon, J.S.; J Nat Prod, 53, 1990,
303-308. 186) Mathuram, V.; Kundu, A.B.; Patra, A.; Seminar on research in Ayurveda
and Siddha CCRAS, New Delhi, March 20-22, 1995. Med Aromat Plant Abstr, 17, 1995, 477.
187) Mathuram, V.; Rao, R.B.; Haldar, S.; Banerji, A.; Kundu, A.B.; J Indian
Chem Soc, 1994, 71, 215- 217.
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188) Sen, A.B.; Singh, S.P.; J Indian Chem Soc, 41, 1964, 192-194. 189) Siuppner, H.; Muller, E.P.; Mathuram, V.; Kundu, A.B.; Phytochemistry
32, 1993, 375- 378.
190) Saxena, R.S.; Gupta, B.; Lata, S.; J Ethnopharmacol, 81, 2002, 321-325.
191) Saxena, R.S.; Gupta, B.; Saxena, K.K.; Srivastava, V.K.; Prasad, D.N.; J
Ethnopharmacol, 19, 1987, 193- 200. 192) Singh, R.C.; Saxena, R.S.; Gupta, B., Saxena, K.K., Prasad, D.N.; Indian
J Pharmacol, 16, 1984, 47. 193) Saxena, R.S.; Gupta, B.; Saxena, K.K.; Singh, R.C.; Prasad, D.N.; J
Ethnopharmacol, 1984, II, 319-330. 194) Paul, B.N.; Saxena, A.K.; J Ethnopharmacol, 56, 1997, 153-158. 195) Devasagayam, T.P.A.; Sainis, K.B.; Indian J Exp Biol, 40, 2002, 639-
655. 196) Singha, U.K.; Guru, P.Y.; Sen, A.B.; Tandon, I.S.; Int J Pharmacog, 30,
1992, 289-295. 197) Misra, P.; Pal, N.L.; Guru, P.Y.; Katiyar, I.C.; Tandon, I.S.; Int J
Pharmacog, 29, 1991, 19-23. 198) Talakal, T.S.; Dwivedi, S.K.; Sharma, S.R.; Pharmaceut Biol, 38, 2000,
326-329. 199) Chitravanshi, V.C.; Singh, A.I.; Ghoshal, S.; Krishna Prasad, B.N.;
Srivastava, V.; Tandon, I.S.; Int J Pharmacog, 30, 1992, 71-75. 200) Goyal, D.K.; Tiwari, S.K.; Agarwal, R.; Sachitra Ayurveda, 51, 1999,
667-668. 201) Sharma, P.V.; Classical Uses of Medicinal Plants. Ist edn. Varanasi:
Chaukhambha Visvabharati (Oriental Publishers & Distributors), 1996, 237-238.
202) Rahman, I.M.F.; Shahriar, M.; Chakraborty, P.; Sattar, M.; Choudhuri,
M.S.K.; Hamdard Med, 43(2), 2000, 19-23.
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