Journal of Ethnopharmacology 82 (2002) 111
/116

www.elsevier.com/locate/jethpharm

Hypoglycemic and antihyperglycemic activity of Murraya koenigii

leaves in diabetic rats
S. Yadav, V. Vats, Y. Dhunnoo, J.K. Grover *
Department of Pharmacology, All India Institute of Medical Science, Ansari Nagar, 110029 New Delhi, India

Received 17 February 2002; received in revised form 8 May 2002; accepted 6 June 2002

Abstract

The commonly used spice curry patta (Murraya koenigii ) is traditionally consumed by diabetics in southern part of India. Feeding
of diet containing various doses of curry leaves (5, 10 and 15%) to normal rats for 7 days as well as mild diabetic (blood glucose
levels
/175 mg/dl induced by alloxan 35 mg/kg IP) and moderate diabetic rats (blood glucose levels
/250 mg/dl induced by STZ
60 mg/kg IP) for 5 weeks showed varying hypoglycemic and anti-hyperglycemic effect. In normal rats, reduction in blood glucose
was almost negligible ( / 4% with 10 and 15% diet). In mild and moderate diabetic rats, feeding of 5, 10 and 15% diet caused a
maximal reduction in blood sugar by 13.1, 16.3 and 21.4% (NS, P B/0.05 and 0.005) and 3.2, 5.58, 8.21% (NS), respectively. The
mechanism of action is further discussed in light of results of previous and the present study. # 2002 Elsevier Science Ireland Ltd.
All rights reserved.

Keywords: Murraya koenigii ; Diabetes; Streptozotocin; Alloxan

1. Introduction resources and a rich history of traditional medicine

Diabetes is the world’s largest endocrine disease
involving metabolic disorders of carbohydrate, fat and
protein. According to WHO projections, the prevalence
of diabetes is likely to increase by 35% (King et al.,
1998). Currently there are over 150 million diabetics
worldwide and this is likely to increase to 300 million or
more by the year 2025 (King et al., 1998; Boyle et al.,
2001). Statistical projection about India suggests that
the number of diabetics will rise from 15 million in 1995
to 57 million in the year 2025 making it the country with
the highest number of diabetics in the world (King et al.,
1998). Therefore, it is necessary to look for new
sloutions to manage this health problem. Although,
many drugs and interventions are available to manage
diabetes, in most instances these are expensive (like
insulin, thiazolidinediones) for a developing country like
India and have adverse effects (like hypoglycemia).
India is a country with a vast reserve of natural
* Corresponding author. Tel.: /91-11-6594897 (O)/4615315 (R);
fax: /91-11-6862663; telex: 31-73042 AIMS IN
E-mail address: jkgrover@hotmail.com (J.K. Grover).
0378-8741/02/$ - see front matter # 2002 Elsevier Science Ireland Ltd. All rights reserved.
PII: S 0 3 7 8 - 8 7 4 1 ( 0 2 ) 0 0 1 6 7 - 8
(Grover and Vats, 2001). Certain spices used commonly
in Indian cooking have been mentioned in these systems
to possess anti-diabetic properties. Murraya koenigii
(MK) is known as ‘Curry Patta’ in Hindi and is widely
use as a spice and condiment in India and other tropical
countries. It belongs to the family Rutaceae (citrus
family). Ayurveda mentions its use as a treatment for
diabetics (Satyavati et al., 1987). Whether these claims
are valid is a subject of great interest and should be
probed scientifically. With this aim we carried out this
study to assess the anti-diabetic property of the leaves of
MK (L. Spengl) in varying degree of hyperglycemia in
rats induced by different doses of alloxan and STZ.
2. Material and methods
2.1. Animals
Animals used for the study were obtained from the
experimental animal facility of the All India Institute of
Medical Sciences after getting approval from Institute
Ethics Committee. Albino rats (both sexes) with body
weight ranging from 150 to 250 g were used for study.
112 S. Yadav et al. / Journal of Ethnopharmacology 82 (2002) 111
/116

Rats were acclimatized for a period of 2
/3 days in the
new environment and subsequently used for further
study.
2.2. Preparation of treatment diet
Fresh green leaves of MK were collected locally (New
Delhi) in the month of July and August and identified
by National Bureau of Plant Genetics Resources and
deposited at the herbarium in National Institute of
Botanical Research against voucher no NBPGR/
NHCP2002-23/600. Leaves were dried under shade at
room temperature ( B/30 8C). After complete drying,
the leaves were grounded into a fine powder using a
domestic electric grinder and mixed thoroughly with the
already grounded standard chow diet. Binder in the
form of 1% starch paste was added to the mixture and
kneaded into a dough with a semi-solid consistency. The
dough was then spread on a tray and cut into pieces of
suitable size and it was then dried at 30 8C in an oven.
Different dose diets were prepared by mixing leaf
powder with chow powder in the ratio of 5, 10 and
15% separately.
2.6. Experimental design for anti-hyperglycemic effect
Rats were divided into 5 groups; normal control
(NC), diabetic control (DC) and treated group MK-5,
MK-10 and MK-15. Control group were fed normal diet
while the treated groups were fed 5, 10 and 15% MK
diet.
2.7. Parameter studied
. Fasting blood glucose: Blood glucose was estimated
by commercially available glucose kit based on
glucose oxidase method (Trinder, 1969) Autopak†
Bayer Diagnostics, Baroda.
. Body weight: Gravimeterically.
2.8. Statistical analysis
Values are given as mean9/S.D. The results were
analyzed by one way ANOVA test using computerized
software, MICROCAL ORIGIN version 2.9, Northampton,
USA.

3. Result
2.3. Preparation of diabetic animals

Administering either alloxan 35 mg/kg or STZ 60 mg/ 3.1. Hypoglycemic effect

kg (IP) produced two different grades of hyperglycemia.
Three days after administration of diabetogenic agents, Basal blood glucose levels of normal control (NC) as
fasting blood glucose was determined and rats with well as MK-5, MK-10, MK-15 were well matched (87
/
more than blood glucose of 175 mg/dl in case of alloxan 89 mg/dl) (see Table 1). After 7 days of treatment, the
and
/250 mg/dl in case of STZ were included in the blood glucose levels in the above-mentioned groups
study. were 87.59/5.97, 84.79/8.54, 81.89/7.47, respectively.
Though the levels decreased, the effect was not sig-
nificant. After the 3rd day of alloxan injection (35 mg/
2.4. Sample collection kg) to the same group of animals, the respective blood
glucose values among treated groups were lower (sig-
Blood samples were collected retro-orbitally from nificant in MK-10 and MK-15) as compared with
inner canthus of eyes using Micro Hematocrit Capil- normal treated group (see Table 1).
laries, Mucaps after at least 12 h of fasted animals.
Table 1
Hypoglycemic and prophylactic anti-hyperglycemic effect of feeding
2.5. Experimental design for hypoglycemic and various doses of MK on glucose levels (mg/dl) in alloxanized rats (35
mg/kg)
prophylactic anti-diabetic effect
Groups Basal Seventh day Tenth day (third day after
Three different diet containing MK leaves powder alloxan 35 mg/kg)
(i.e. 5, 10 and 15%) were fed to normal rats for 1 week.
DC 8796.30 87.595.97 175.5910.87
On the 7th day, blood sugar was estimated to assess the MK-5 86.697.24 84.798.54 173.796.71
hypoglycemic effect of the plant diet. The same rats were MK-10 86.7598.86 81.897.47 169.2910.56*
then injected with alloxan (35 mg/kg body weight IP) MK-15 86.197.43 81.897.47 162.3911.23*
and plant diet was continued till the next 3 days. Blood
Values are given as mean9S.D. for eight animal each. DC, Diabetic
sugar was estimated on the last day of the experiment control; MK, Murraya koeingii . Treated groups were compared with
(i.e. 10th day) to assess the prophylactic antidiabetic corresponding value of the diabetic control. Values were singnificant at
effect on development of hyperglycemia. *P B 0.05.
 S. Yadav et al. / Journal of Ethnopharmacology 82 (2002) 111
/116 113

Table 2
Effects of feeding various doses of MK on blood glucose levels (mg/dl) in alloxanized rats (35 mg/kg)

Basal Third day First week Second week Third week Fourth week Fifth week

NC 86.5910.21 83.5911.6 84.5913.32 85.5911.92 84.7912.41 85.7910.08 84.7913.32

DC 82.797.94 200.6928.88*** 200.2930.25*** 201929.59 (1.49)*** 200.7927.30*** 200.5927.01*** 200.6926.65
(n 7)***
MK-5 83.899.73 202.7925.32 199.3924.61 (1.66) 195.3924.55 (3.63) 191.6925.16 (5.48) 186.6925.41 (7.95) 176.1918.60
(n 7)
MK-10 86.1912.86 1.99.1924.32 193.3923.02 (2.88) 187.8923.32 (5.64) 182.7923.18 (8.22) 177.5924.34 (10.86) 166.5917.95
(n 7)*
MK-15 85.297.64 204.7928.97 199.3925.69 (2.62) 192.7925.84 (5.86) 186.3925.61 (8.97) 174.7920.82 (14.65) 160.7914.87
(n 7)**

Values are given as mean9S.D. for eight animals in all case except where indicated individually. Abbrevations as in Table 1. Values were
significant at *P B 0.05; **P B 0.005; ***P B 0.0005.

3.2. Antihyperglycemic studies in alloxanized (35 mg/kg)
rats
The basal blood glucose levels of all groups were
statistically not different from each other (see Table 2).
Three days after alloxan administration, blood glucose
values were 2
/5-folds higher (P B/0.0005) in all the
groups in comparison to NC and were not statistically
different from each other. Value of blood glucose at 1st,
2nd, 3rd and 4th week decreased in all the treatment
groups but remained stable in diabetic controls. Highest
reduction in blood glucose was seen in MK-15. The
percentage reduction was dose dependent in the MK
groups animals. However, this reduction was not
statistically significant. The reduction in blood glucose
at 5th week was statistically significant in MK-10 (P B/
0.05) and MK-15 (P B/0.005) treated groups. Plateau
effect was not reached in either treatment.
3.3. Antihyperglycemic studies in STZ diabetic rats (60
mg/kg)
All groups of rats had nearly same blood glucose
values at basal level and were not significantly different
from each other. Seventy-two hours after administration
of STZ, blood glucose values were significantly raised by
more than 3-folds and the rise was same in all the
groups. Blood glucose values decreased only marginally
after 1, 2, 3, 4 and 5 weeks of treatments. However, this
reduction was insignificant in comparison to the diabetic
controls at all time periods in the treated groups (see
Table 3). Maximal reduction of 3.2% by MK-5 and
5.58% by MK-10 was seen at 5th week. However, it was
8.21% in MK-15 at 4th week of treatment.
3.4. Body weight in alloxanized rats
The basal body weight (mean) of all groups ranged
from 193 to 199 g and there was no inter-group
variation. Administration of alloxan led to a loss in
body weight after 1 week of treatment but this was
statistically insignificant. On the other hand, normal
controls showed an increase in body weight by 7% at the
end of the experiment (i.e. 5th week). The body weight
of diabetic controls was significantly less at the end of
3rd week (P B/0.05), 4th week (P B/0.05) and 5th week
(P B/0.005) in comparison to NC. Though the body
weights of the treated groups were increased, it was not
significantly different from the diabetic controls (see
Table 4).
3.5. Body weight in STZ rats
Basal body weights of all groups ranged from 197 to
203 g and were not significantly different. After 1 week
of treatment, body weight decreased by 3
/5% in all

Table 3
Effects of feeding various doses of MK on blood glucose levels (mg/dl) in STZ rats (60 mg/kg)

Basal Third day First week Second week Third week Fourth week Fifth week

NC 86.5910.21 85.3913.02 84.5913.30 85.6910.52 83.7916.50 8399.81 84.298.73

DC 82.797.94 275.3965.31* 276.5964.13* 275.3965.24* 275.5964.38* 274.6962.91* 271.8963.11 (1.4)*
MK-5 83.899.73 287.6931.66 285.6931.36 (0.69) 285.5931.03 (0.73) 282.2932.34 (1.86) 279.7932.83 (2.73) 278.2931.98 (3.2)
MK-10 86.1912.86 290.8934.51 288.3934.52 (0.85) 285.2933.82 (1.93) 281.3932.14 (3.26) 277.2932.01 (4.68) 274.6932.36 (5.58)
MK-15 85.297.64 283954.51 290.5961.30 (1.76) 272.8953.74 (3.57) 267.8954.39 (5.34) 259.7953.12 (8.21) 260.3952.09 (7.99)

Values are given as mean9S.D. for eight animals eight. Abbrevations as in Table 1. Values were significant at *P B 0.0005.
114 S. Yadav et al. / Journal of Ethnopharmacology 82 (2002) 111
/116

Table 4
Effect of feeding various doses of MK on body weight (g) in alloxanized rats (35 mg/kg)

Basal First week Second week Third week Fourth week Fifth week

NC 199.8913.24 202.5912.71 205.2912.63 207.5912.69 212.3911.79 214.7912.11

DC 195.296.05 191.1915.61 192915.21 192.2914.19* 192.6913.50* 193.6912.7**
MK-5 197.1915.55 192.7915.92 194.5915.41 196915.53 197.1915.30 199.5915.83 (n 7)
MK-10 194.2913.22 189.3912.82 191.3912.83 193.1912.86 195913.80 197915.50 (n 7)
MK-15 193915.60 188.7915.21 190.6915.52 192.7915.62 195.2916.68 196.5916.93 (n 7)

Values are given as mean9S.D. for 8 animals in all cases except where indicated individually. Abbreviations as in Table 1. Values were significant
at *P B 0.05, **P B 0.005.

groups except the normal controls but this decrease was
not significantly different. Normal controls showed
weight gain by 1.47, 3.44, 4.43, 5.9 and 7.3% at 1st,
2nd, 3rd, 4th and 5th week of treatment, respectively.
On the other hand, body weight in diabetic controls
showed continuous decrease throughout the experimen-
tal period and remained significantly less than the
normal controls (P B/0.05 at 2nd and 3rd week and
P B/0.005 at 4th and 5th week). The body weight lost in
the 1st week was regained in the treated groups and
reached the basal values at 5th week (198.69/16.33,
201.39/13.26, 202.19/18.12 in MK-5, MK-10 and MK-
15). However, they remained significantly less in com-
parison with normal controls (see Table 5).
4. Discussion
Historical accounts reveal that knowledge regarding
diabetes existed since Brahmic period as this was
mentioned in the Ayurvedic textbooks-Sushruta Sam-
hita-written in fourth and fifth centuries B.C. (Dhanu-
kar and Thatte, 1989). In this ancient text, two forms of
diabetes were described; one genetically based and the
other as a result of dietary indiscretion (Dhanukar and
Thatte, 1989). Even the treatment in the Indian ancient
pharmacopoeia mentioned specific treatments for the
two types including dietary modifications, medicinal
plant remedies and minerals. Moreover the research
conducted over last several decades has shown plant and
plant based therapies have a potential to control and
treat diabetes (Oliver Bever and Zahnd, 1979; Bailey and
Day, 1989; Ivorra et al., 1989; Marles and Farnsworth,
1995) and its complications (Grover et al., 2001). Role
of Indian plants as an anti-diabetic has been reviewed
elsewhere (Grover et al., 2002).
Appropriate nutritional management is essential for
restoring and maintaining a normal metabolic state.
Therefore, diet remains a cornerstone in diabetic man-
agement. Spices and cereals like Fenugreek, MK,
Brassica juncea as dietary constituents have been found
to have beneficial effect on carbohydrate metabolism
experimentally as well as clinically (Iyer and Mani, 1990;
Vats et al., 2002; Khan et al., 1995).
In the present study we have investigated the efficacy
of feeding curry leaves (MK) as dietary constituents in
controlling hyperglycemia. Feeding of diet containing
various doses of MK to the normal rats for 7 days did
not show any profound hypoglycemia in this study and
this is in contrast to the report of Khan et al., 1995. This
may be possibly due to shorter duration (7 days) of the
present study in comparison to 60-day duration in the
Khan study. However, MK exerted profound anti-
alloxan effect as diet containing higher proportion of
MK significantly prevented the rise in blood glucose
after administration of alloxan (35 mg/kg).
Alloxan and STZ produce hyperglycemia by selective
cytotoxic effect on pancreatic b cells. One of the

Table 5
Effect of feeding various doses of MK on body weight (g) in STZ rats (60 mg/kg)

Basal First week Second week Third week Fourth week Fifth week

NC 203.7919.77 206.8919.87 210.1919.05 212.8918.68 215.7918.32 218.2918.12

DC 197.5913.35 190.7913.01 189.5912.99 188.2913.41* 188.8912.50* 189.3912.37**
MK-5 200.7915.26 194.2914.58 195.2914.77 196.1915.42 197.5916.02 198.6916.33
MK-10 200.111.59 193.5912.03 195.3911.83 197.2912.30 199.7912.84 201.3913.26
MK-15 199918.72 193918.43 195.2917.90 197.6917.73 199.7917.65 202.1918.12

Values are given as mean9S.D. for eight animals each. Abbreviations as in Table 1.
 S. Yadav et al. / Journal of Ethnopharmacology 82 (2002) 111
/116
intracellular phenomenon for its cytotoxicity is through
generation of free radicals demonstrated both in vivo
and in vitro (Gandy et al., 1982; Papaccio et al., 1986).
MK probably prevented the destruction of b cells of
islets in the pancreas. This is an interesting finding and
suggests that it may have antioxidant or free radical
scavenger properties in preventing these changes. Anti-
oxidant/free radical scavenging property of MK leaves
has been stated earlier by many authors (Tachibana et
al., 2001; Khan et al., 1996, 1997; Ramsewak et al.,
1999). Thus MK may have a role in prevention of
diabetes and its consumption should be encouraged in
the early diabetic state.
MK showed moderate antihyperglycemic activity in
mild diabetic rats (induced by alloxan 35 mg/kg). The
onset of antihyperglycemic effect was seen in a week’s
time though the maximum and statistically significant
effect was observed only after several weeks of feeding
these dietary agents. The antihyperglycemic effect con-
tinued to increase with duration. The plateau, however,
was not achieved even after 5 weeks indicating a
possibility of achieving normal blood sugar on a later
day. Moreover as has been reported earlier (Grover et
al., 2000), it is important that to obtain maximum effect
therapy with plant products be continued for longer
duration. Unlike the sulfonylureas and biguanides, the
onset of action is slow and it cannot be used for
management of emergency situations.
No significant antihyperglycemic effect was seen in
moderate diabetes (induced by STZ 60 mg/kg). A
plateau effect was seen with higher doses of MK by
the end of 3rd or 4th week indicating that MK is not of
utility in the management of moderate or severe variety
of diabetes though a clinical study by Iyer and Mani
(1990) involving 30 type II patients reported a signifi-
cant reduction (P B/0.05) in fasting blood sugar on 15th
day (12 gm curry leaves powder per day orally).
The mechanism for hypoglycemia activity of MK as
demonstrated by Khan et al. (1995) is through increased
utilization of glucose. However, based on the results of
the present study, the probable mechanism seems to be
the release of insulin as MK displayed moderate
antihyperglycemic activity in mild diabetics but poor
response in moderate diabetics.
Other parameters of diabetes such as body weight
were also affected on treatment with MK. There was
gradual increase in body weight in the normal controls
while the diabetic controls continued to lose weight.
This is in agreement with earlier findings of Prince et al.
(1998). Treatments with MK decreased the reduction in
body weight by diabetes. The treatments altered the
body weights of diabetic animals towards normalcy.
Thus, in conclusion of above-mentioned facts, the
present study suggests that MK can play role in
management of pre-diabetic state or mild diabetes. For
control of moderate and severe diabetes this agent alone
115
is not likely to be useful and may be of no use in type I
diabetes. The spice seems to be safe in long-term use as it
has been safely consumed through ages as a dietary
constituent.
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