Evidence Based Ayurveda For Revitalization of Mental Health

Volume 01
Number 02
Page 51 - 99
December 2011
ISSN 2012 9238
Sri Lanka Journal of Indigenous Medicine
SLJIM
Peer reviewed research publication of the
INSTITUTE OF INDIGENOUS MEDICINE

University of Colombo, Rajagiriya, Sri Lanka
Sri Lanka Journal of Indigenous Medicine (SLJIM)

Volume 01 Number 02 Page 51 - 99 December 2011
EDITORIAL BOARD
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Kaumarabhrithya
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Department of Zoology, Faculty of Science, University of
Colombo
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Cover story
Bacopa monnieri (L) Pennel (Scrophulariaceae)
Sinhala: Lunuwila; Tamil: Pirami; Hindi: Brahmi;
English: Thyme-leaved gratiola
Brahmi or Lunuwila possesses numerous
medicinal properties. Its uses are documented in
ancient Ayurvedic texts and the herb has been
widely used to promote the intellect, and treat
neurological and mental problems. This plant is
commonly distributed in moist and damp areas
on the edges of streams and water trenches. It is
a prostrate, glabrous and fleshy herb. The leaves
are sessile, soft, and succulent up to 2.5 mm long
with obscure venation. The stem is 10-30 cm long
and 1-2 mm thick, with soft ascending branches.
Flowers white or blue with purple veins, axillary
and solitary on peduncles usually longer than
the leaves. Fruits ovoid, acute capsules include
in the persistent calyx.
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Brahmine and Herpestine are major alkaloid

present in the aerial parts. Flavonoid such as
glucuronyl-7-apigenin and glucuronyl-7-luteolin
are present. Bacosides and Bacosaponins are
important saponin constituents.
Brahmi has the capacity to improve the higher
order cognitive processes and improve learning
capability. It also has anxiolytic activity, anti
depressant activity, intellect promoting activity
antioxidant property, analgesic activity,
spasmolitic activity, and bronchodilatory activity.
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ISSN 2012-9238

Contents
Original Papers
Experimental evaluation of gastroprotective and adaptogenic activity of
Amalakayas Rasayana and its vehicle (ghee and honey)
51
S M S Samarakoon, S K M K Herapathdeniya, H M Chandola, B Ravishankar
Study of the efficacy of an ayurvedic treatment regimen on balaka pakshaghatha

with special reference to cerebral palsy
55
Saroja Weerakoon, A P G Amarasinghe
Clinical efficacy of Dashamoola Taila Matra Vasti on management of primary dysmenorrhoea
59
Kaumadi Karunagoda, Shilpa Donga, Lakshmi Priya Dei
In vitro evaluation of different aqueous extracts of Senna alata leaves for antibacterial activity
64
E Christy Jeyaseelan, S Tharmila, A C Thavaranjit
Selection of the most suitable pot height and harvesting stage for higher growth,
yield and oil quality of Vettiver (Vetiveria zizanioides)
70
N D N Priyadarshani, M K T K Amarasinghe, S Subasinghe, I R Palihakkara, H K M S Kumarasinghe
Anti hyperlipidemic effect of Vara Asanadi Kwatha against high fat diet induced hyperlipidemic rats
76
Anju P Ramachandran, M Shyam Prasad, Vijay Kumar, B K Ashok, B Ravishankar, H M Chandola
Short Communication
Antibacterial properties of Accmus mouth wash
83
S Tharmila, T Thileepan, A C Thavaranjit, R Srikaran
Review Papers
Anti rheumatic herbal compound drug Yi Shen Juan Bi (YJB) as selective cytokines
target in rheumatoid arthritis
86
Pathirage Kamal Perera, Yunman Li
Evidence based Ayurveda for revitalization of mental health
91
Nisha Ojha, Abhimanyu Kumar
Guidelines for authors
98
Original Paper
51
Experimental evaluation of gastroprotective and adaptogenic activity

of Amalakayas Rasayana and its vehicle (ghee and honey)
S M S Samarakoon1, S K M K Herapathdeniya2, H M Chandola3, B Ravishankar4
Abstract
Amalakayas Rasayana (AR) was tested for its antiulcer activity in forced swimming induced hypothermia
and stress induced gastric ulceration. AR was
administered in the dose of 270 mg/kg orally for 7
consecutive days prior to the experiment. The
adaptogenic activity was assessed by determining and
comparing the changes in rectal temperature and ulcer
index and compared in AR and vehicle treated group
against stress control group. In forced swimming
induced gastric ulceration, pretreatment with AR caused
significant attenuation of ulcer index when compared with
both stress control (p<0.001) and vehicle control (p<0.05)
groups. AR exhibits significant reduction in ulcer index in
comparison to stress control group (p<0.05) and vehicle
control (p<0.001) groups. The results suggest that AR
possesses significant adaptogenic and gastro protective
activities.
Introduction
Ageing is the accumulation of changes in humans
refers to a multi-dimensional process of physical,
psychological, and social changes [1]. The ageing process
is a biological reality which has its own dynamic course
that is beyond human control. Ageing is defined as a
progressive generalized impairment of function resulting
in a loss of adaptive response to stress and in a growing
risk of age associated disease [2].
Ayurveda has classified ageing into two viz., Kalaja
jara (Physiological ageing) which is natural process of
ageing and Akalaja jara (premature ageing) [3]. Ayurveda
has described various rejuvenative therapies with the help
of special class of medicinal preparations called Rasayana
that are believed to rebuild the body, mind, prevent degeneration and postpone ageing [4]. Amalakayas Rasayana
(AR) is one among many Rasayana formulations
mentioned in Ayurvedic classical text Charaka Samhita for
the treatment of ageing related disorders [5] and used by
Ayurvedic physicians. However, no report on the

pharmacological screening on this formulation is available;
hence, this study was designed to assess adaptogenic
and anti-stress activities of AR to provide pharmacological
basis to clinical claims and to justify its use as anti-ageing
medicine.
Materials and Methods

Test drug: The raw materials (Table 1) of the test formulation
were collected from Gujarat Ayurveda University pharmacy
and were subjected to pharmacognostical studies in order
to establish their authenticity. From the raw materials, the
test drug AR was prepared following the classical
guidelines [6]. The vehicle viz., honey and ghee of standard
brands were purchased from the local market.
Chemicals: All the chemicals and reagents used in the
experimental study were procured from standard and
reputed firms and are of analytical grade regularly being
used in the laboratory.
Animals: Charles Foster strain albino rats of either sex
weighing between 200 30g were selected and procured
from the animal house attached to the institute (Registration
No.548/2002/CPCSEA). They were housed in large
spacious polypropylene cages and fed with Amrut brand
rat pellet feed supplied by Pranav Agro Industries and tap
water given ad libitum. The animals were acclimatized for
at least one week in lab condition before commencement
of the experiment in standard laboratory conditions 12
01 hour day and night rhythm, maintained at 25 3C and
40 to 60% humidity. Before the test, the animals were kept
fasting for 12 hours. Institutional Animal Ethics Committee
had approved the experimental protocol (Approval
number: IAEC 05/09-10/Ph.D.08) and the care of animals
was taken as per the CPCSEA Guidelines (Committee for
the Purpose of Control and Supervision on Experiments
on Animals) [7].
Department of Kayachikitsa, Gampaha Wickramarachchi Ayurveda Institute, University of Kelaniya, Yakkala, Sri
Lanka.
2
Department of Dravyagunavignana, Institute of Indigenous Medicine, University of Colombo, Rajagiriya, Sri Lanka.
3
Department of Kayachikitsa, Institute of Postgraduate Teaching and Research in Ayurveda, Gujarat Ayurved University,
Jamnagar, India.
4
Pharmacology Laboratory, Institute of Postgraduate Teaching and Research in Ayurveda, Gujarat Ayurved University,
Jamnagar, India.
Correspondence: Dr. S. M. S. Samarakoon, Senior Lecturer, Department of Kayachikitsa, Gampaha Wickramarachchi
Ayurveda Institute, University of Kelaniya, Yakkala, Sri Lanka. E-mail: samarakoonsms@yahoo.com. Received 04
August and revised version accepted 12 November 2011.
Samarakoon et al. Experimental evaluation of gastro protectiveSLJIM 2011; 01 (02): 51-54
52
Original Paper
Table 1: Formulation composition of Amalakayas Rasayana
Ingredients
Botanical Name
Part used
Quantity
Amalakai
Emblica officinalis
Fruit
11 Part
Shweta
Alpenia galanga
Rhizome
1 Part
Shatavari
Asparagus racemosus
Root
1 Part
Punarnava
Boerhavia diffusa
Root
1 Part
Manduka parni
Centella asciatica
Whole plant
1 Part
Shalaparni
Desmodium gangiticum
Root
1 Part
Jivanti
Leptadenia reticulate
Root
1 Part
Rasna
Pluchea lanceolata
Root
1 Part
Haritaki
Terminalia chebula
Fruit
1 Part
10
Guduchi
Tinospora cordifolia
Stem
1 Part
11
Lauha Bhasma
Incinerated Iron
1.5 Part
Dose selection and schedule: The classical dose of AR in

human beings is 3 g/day [8]. The dose for experimental
animals was calculated by extrapolating the human dose
to animals (270 mg/kg) based on the body surface area
ratio by referring to the standard table of Paget and Barnes
(1964) [9]. The drug solution was made by adding unequal
quantity of ghee (700 mg/kg) and honey (1350 mg/kg) as
per the classical indication [10] and administered to animals
orally with the help of gastric catheter sleeved to syringe.
The drugs were administered to over night fasted animals.
Adaptogenic and anti-ulcer activity [11]: The selected
animals were divided in to four groups of six animals in
each. Normal control (water control WC) animals were
kept under standard laboratory conditions, left undisturbed
in their home cages without stress exposures. Second
group received only distilled water and served as stress
control (SC) group where as the third group received
combination of ghee (700 mg/kg) and honey (1350 g/kg)
and served as vehicle control (VC). Fourth group (AR)
was administered with the AR (270 mg/kg) plus vehicle.
For the experimental group, drugs were given for seven
consecutive days. On sixth day the rats were kept in
individual metabolic cages to prevent coprophagy and
fasted for 36 hours with access to water ad libitum. On
the seventh day one hour after drug administration, the
initial rectal temperature of individual rats was noted. After
noting initial rectal temperature rats are kept inside
specially arranged containers, which were made up of
plexiglass with holed lids. The water level was maintained
up to 25 cm height and temperature of water was maintained
at 22 2 C. Rats were placed in the container and exactly
after 20 minutes of exposure to stressed condition, the
rats were taken out individually and final rectal temperature
of each rat was noted. The drop in rectal temperature was
noted down.
Effect of drugs on stress-induced ulcer was evaluated

by following the method of Parmar and Jagruti [12]. which
was modified according to the experimental need. The rats
after noting their final rectal temperature were again
exposed to the swimming stress inside the same container
for 16 hours. At the end of 16 hour period blood was
obtained from the retro-orbital puncture under light ether
anaesthesia using capillary tubes. The body weight was
noted and then they were sacrificed. Blood samples were
collected for assessing different types of haematological
parameters by using automatic haematological analyzer
(ACRUS automated haematology auto-analyzer). The vital
organs like liver, heart, kidney and adrenals were dissected
out, cleaned for extraneous tissues, blotted with tissue
paper, weighed and computed per 100 g body weight.
The stomach was excised, cleaned and opened along
the greater curvature. The inner surface was cleaned gently
by washing with cold saline solution and spread on wax
board with the mucous surface upwards avoiding
corrugation and examined for ulceration with a magnifying
lens. Severity of ulcer and total number of ulcers in each
rat was recorded for calculating ulcer index. Ulcer index
was calculated according to the method described by
Kulkarni and Goel [13]. Mean ulcer scores for each
experimental group were calculated and expressed as the
ulcer index.
Statistical analysis: The results were presented as mean
SEM for six rats in each group. Statistical comparisons
were performed by unpaired students t test and one way
Anova with Dunnets multiple t test as post-hoc test by
using Sigma Stat Software (version 3.1) and the level of
significance was set at p<0.05.
Original Paper
Results
Effect of AR on rectal temperature
Table 2: Effect of AR and vehicle on rectal temperature
in rats subjected to forced swimming stress
Group Dose (g/kg) Percentage decrease %
in rectal
change
temperature (C)
SC
QS
28.046 0.835
--
VC
0.7+1.4
22.608 1.909 *
19.39
AR
0.27+0.7+1.4 16.470 0.530 **
41.32
*p<0.05 **p<0.001 Vs stress control (unpaired t test); p<0.05 Vs

vehicle control (unpaired t test)
Administration of vehicle and test drug significantly

decreased rectal temperature in comparison to stress
control group. Further, test drug shows statistically
significant decrease in rectal temperature in comparison
to vehicle control group (Table 2).
Effect of AR and vehicle on ulcer index

Table 3: Effect of AR and vehicle on ulcer index in rats
subjected to forced swimming stress
Group
Dose (g/kg)
Ulcer index
% change
SC
QS
47.58 2.44
VC
0.7+1.4
20.70 2.55#
56.49
AR
0.27+0.7+1.4
05.90 0.48#*
87.60
# One
way Anova - F value 102.50; p<0.001: p<0.05 for VC and

AR Vs stress control.
* p<0.001
Vs vehicle control (Unpaired t test)
Administration of vehicle and test drug significantly

attenuated stress induced ulceration in comparison
to stress control group. Further, test drug shows
statistically highly significant decrease in ulcer index in
comparison to vehicle control group (Table 3).
53
promotion of health by revitalizing the metabolism and

enhancing immunity. Rasayana therapy encompasses
procedures of revitalization and rejuvenation to increase
the bodys power of resistance to disease and supposed
to slowdown the advancement of ageing [15].
Swimming stress in small laboratory animals has been
widely used for studying the physiological changes and
the capacity of the organism to adjust in response to stress
[16]. Swimming is not always a simple exercise stress,
because emotional factors are difficult to be eliminated.
Even short single stress like one day forced swimming
stress is as effective as prolonged stressor in bringing
about the stress induced alterations in the body [17].
Swimming induced hypothermia is an inevitable outcome
of swimming at water temperature lower than the animals
core temperature. In this study, forced swimming lead
to remarkable hypothermia and pre-treatment with both
vehicle (p<0.05) and AR (p<0.001) attenuated it in
significant manner. The magnitude of attenuation
observed in AR treated group is comparatively high in
comparison to vehicle (p<0.05). Thus, the observed adaptogenic effect can mainly be due to adaptogenic
properties of AR.
Stress ulcers are due to both physiological and
psychological factors, which is crucial for gastrointestinal
defense and increased accumulation of acid and pepsin
leading to auto-digestion of the gastric mucosa [18]. Stress
in animals is known to increase gastric motility and acidity
which could lead to ulceration manifested by severe
mucosal damage and haemorrhage [19]. Importance of
impaired mucosal blood flow also appears among the
important factors in the pathogenesis of stress-induced
ulcers [20].
The other factors that may be involved are plateletactivating factor [21], increase in gastric motility, vagal
over activity [22], mast cell degranulation [23] and
decreased prostaglandin (PG) synthesis [24]. The reactive
oxygen species generated by the metabolism of
arachidonic acid, platelets, macrophages, and smooth
muscle cells may also contribute to gastric mucosal
damage [25]. Results presented in this work showed that
oral administration of AR and vehicle before stress
induction decreased the incidence and severity of stress
induced gastric ulcers in rats (p<0.05). Attenuation of ulcer
index in pretreated with AR against vehicle control group
is highly significant (p<0.001).
Conclusion
Discussion
Ageing is universal but complex biological process
with definite manifestations characterized by impairment
of various functions and decreased ability to respond to
stress [14]. Rasayana chikitsa is a specialized section of
Ayurveda, which mainly deals with the preservation and
Many of the drugs in AR are reported to have antistress and adaptogenic activity. Further the vehicle;
combination of ghee and honey is also reported to have
adaptogenic activity. Thus, the observed adaptogenic
profile of AR may be attributed to one or more bioactive
principles present in these drugs. From this study, it can
54
Original Paper
be concluded that AR is having significant adaptogenic

and gastroprotective activity. The observed adaptogenic
and anti-stress effect may probably be either through
attenuation of stress induced stimulation of hypothalamus-pituitary-axis (HPA), quenching of free radicals,
and enhancement of cell proliferation or cellular
detoxification mechanisms.
References
1.
Kirkwood T, Ebrahim S, Kalache A. Mechanisms of Ageing

in Epidemiology in Old Age, BMJ Publishing Group,
London. 1996; 3.
2.
Hamilton IS. The Psychology of Ageing: An Introduction,

Jessica Kingley Publishers, London. 1998; 1.
3.
Acharya JT. Sushruta Samhita, Chaukmbha Surabharati

Prakashan, Varanasi. 2008; 8.
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Acharya JT. Charaka Samhita, Chaukambha Prakashan,

Varanasi. 2004; 377.
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Samarakoon SMS, Chandola HM, Ravishankar B. A clinicexperimental study on the efficacy of Amalakayas Rasayana
in the management of premature ageing, PhD thesis, Gujarat
Ayurveda University, Jamnagar. 2010; 139-67.
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Anonymous. Ayurvedic Formulary of India, Department

of AYUSH, New Delhi, 2007; Part-I: 96.
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Paget GE, Barnes JM. Evaluation of drug activities.

Pharmacometrics, Academic Press, New York. 1964; 1:
161.
10. Acharya JT. Charaka Samhita, Chaukambha Bharati

Academy, Varanasi. 2004; 520.
11. Sheth MD, Rege NN, Dahanukar SA. Effect of Tinospora
cordifolia on gastrointestinal dysmotility induced by
chronic, unpredictable wrap-restraint. Indian Journal of
Pharmacology 2001; 331-5.
12. Parmar NS, Jagruti KD. A review of the current methodology
for the gastric and duodenal anti-ulcer agents. Indian Journal
of Pharmacology 1993; 25: 120-35.
13. Kulkarni SK, Goel RK. Gastric antiulcer activity of UL409 in rats. Indian J Exp Biol 1996; 34: 683-8.
14. Samarakoon SMS, Chandola HM, Ravishankar B, Ashok
BK, Gupta Varun B. Evaluation of an adaptogenic activity
profile of a compound Ayurvedic formulation Amalakayas
Rasayana. Indian Journal of Traditional Knowledge 2011;
10(4): 661-7.
15. Handa SS. Rasayana drugs. Pharmatimes 1993; Part-I: 914.
16. Gehlot A, Godhwani JL, Godhwani S, Aseri ML, Jain P,
Vyas MCR. Sound stress-induced changes and their
modification by drugs in albino rats an experimental study.
Indian Journal of Pharmacology 1997; 29: 187-9.
17. Nagaraja HS, Jaganathan PS. Forced swimming stress
induced changes in the physiological and biochemical
parameters in albino rats. Indian J Physiol Pharmacol 1999;
43(1): 53-9.
18. Goel RK, Bhattacharya SK. Gastroduodenal mucosal
defense and mucosal protective agents. Indian Journal of
Experimental Biology 1991; 29: 701-14.
19. Salim AS. The significance of removing oxygen derived free
radicals in the treatment of acute and chronic duodenal
ulceration in rats. J Pharm Pharmacol 1990; 42: 64-7.
20. Schiessel R, Feil W, Wenzel E. Mechanisms of stress
ulceration and implications for treatment. Gastroenterology
Clinics of North America 1990; 19: 101-20.
21. Cho CH. The role of endogenous ulcerogenic mediators in
the pathogenesis of peptic ulcer. Life Sciences 1994; 55:
1521-35.
22. Cho CH, Ogle CW, Dai S. Acute gastric ulcer formation in
response to electric vagal stimulation in rats. European
Journal of Pharmacology 1976; 35: 215-9.
23. Cho CH, Ogle CW. Cholinergic mediated gastric mast cell
degranulation with subsequent histamine H1 and H2
receptor activation in stress ulceration in rats. European
24. Miller TA. Mechanism of stress-related mucosal damage.
American Journal of Medicine 1987; 83: 8-14.
25. Repetto MG, Llesuy SF. Antioxidant properties of natural
compounds used in popular medicine for gastric ulcers.
Brazilian Journal of Medical and Biological Research
2002; 35(5): 523-34.
Original Paper
55
Study of the efficacy of an ayurvedic treatment regimen on Balaka

Pakshaghatha with special reference to cerebral palsy
Saroja Weerakoon1, A P G Amarasinghe1
Abstract
Balaka Pakshaghatha (BP) is a condition that affects
movement, posture and co-ordination, and it is caused
by Shiromarmabhighata (damage of the brain), before,
during or soon after birth. Because of this problem most
of the children have to face many motor activities
dysfunction and it becomes a common developmental
disability problem. Cerebral palsy (CP) is also a condition,
described in modern medical science resulting from
damage to the brain before, during or soon after birth.
Etiopathogenisis and symptoms of Balaka Pakshaghatha are similar to cerebral palsy. Objectives of this
study were to evaluate the efficacy of an Ayurvedic
treatment regimen with selected Pancha Karma (bio
purification measures) treatments for the management
of Balaka Pakshaghatha with special reference to
cerebral palsy and to compare the effectiveness of
Ayurvedic treatment and Physiotherapy treatment for the
management of Balaka Pakshaghatha. Sixty patients in
the age group of 1-6 years of CP/BP were taken for the
study from the Paediactric Clinic of Ayurveda Teaching
Hospital, Colombo and Physiotherapy Unit of
Awissawella Base Hospital in Sri Lanka. Sixty patients
were divided in to two groups, 30 patients in each group.
The test group of 30 patients were selected for Ayurvedic
treatment. 30 patients were randomly selected for
physiotherapy treatments. Ayurvedic treatment regimen
was of three rounds of treatment of 45 days in each round
with two months gap and six months follow up. The
efficacy of each therapy was evaluated by Gross Motor
Function Classification System. Both Ayurvedic and
Physiotherapy treatments had significant result of p value
at < 0.05. In Ayurvedic, treatment p value is 9.44 E-13 and
in the Physiotherapy treatment, has been 3.32 E-08.
Ayurvedic treatment regimen and Physiothrapy have the
capacity to improve the gross motor functions of Balaka
Pakshaghatha. However, Ayurvedic treatment regimen
is highly effective than Physiotherapy treatment in the
management of Balaka Pakshaghatha.
Introduction
Balaka Pakshagatha (BP) or Cerebral Palsy (CP) is a
condition that affects movement, posture and coordination, and it is caused by damage to the brain before,
during or soon after birth [1]. Most of the children have
to face many motor activity dysfunctions due to this
problem and it becomes a common developmental
disability problem. It has been recognized as one of the
Vatha vyadhi in the field of Ayurvedic medicine [2].
Annual incidence of BP/CP is 2-2.5 per 1000 births [3]
Extrapolated statistics for Balaka Pakshaghatha in Sri
Lanka is 39810 per 19, 905, 165 [4]. The objectives of
this study were to evaluate the efficacy of an Ayurvedic
treatment regimen with selected Pancha Karma (Bio
Purification measures) treatments available in Paediatrics
Ward at Boralla Ayurvedic Teaching Hospital for the
management of Balaka Pakshaghatha with special
reference to Cerebral Palsy and to compare the
effectiveness of Ayurvedic treatment and Physiotherapy
treatment for the management of Balaka Pakshaghatha.

Sixty patients in the age group of 1-6 years of BP/CP
were taken for the study irrespective of their sex, race and
religion etc. from the Paediactric Clinic of Ayurvedic
Teaching Hospital Colombo and Physiotherapy Unit in
Awissawella base Hospital.
Patients with history of delayed milestone, spasticity
in one or all the limbs, persistence of primitive reflexes
were diagnosed by using performa as BP/CP and
selected for the study. Children who have history of
delayed developmental milestone, body stiffness, communication difficulties, restless behaviour, abnormal reflexes,
BP/CP with history of convulsion were included in this
present study. Children those who are suffering from
convulsion, who are not in progress after two months
OPD treatment, children below one year and above 6 years
of age, and those who have other systemic disorders like
Asthmatic condition, Heart disease etc. were excluded.
The sample of sixty patients was divided in to two
groups, including 30 patients of each group. Test Group
Department of Prasutitantra Kaumarabhrithya, Institute of Indigenous Medicine, University of Colombo,

Correspondance: Dr. Saroja Weerakoon, Lecturer, Department of Prasutitantra Kaumarabhrithya, Institute of
Indigenous Medicine, University of Colombo, Rajagiriya, Sri Lanka. E-mail: sarojaweerakoon@yahoo.com.
Received 24 August and revised version accepted 16 November 2011.
Saroja and Amarasinghe. Ayurvedic Treatment Regimen on Balaka PakshaghathaSLJIM 2011; 01(02): 55-58
56
Original Paper
(Group A), who had some progress after two months

O.P.D. treatments, were selected for Ayurvedic
Treatment. Patients in Phsiotherapy group (Group B) were
selected considering their age range such as 1-2 years,
2-4 years and 4-6 years for Physiotherapy Treatments.
Ayurvedic treatment schedule
Outdoor treatment period was two months.The
patients who had some progress were treated in Indoor
Patient Department with three rounds of treatments of
45 days, each round with two months gap, and six months
follow up.
Methods of administration of drugs
In first two weeks Patients were treated with
Etamadeduru Decoction [5] and Bhrahmi Mandukaparnie
Maduyashti Decoction (Bacopa monnieri L., Centella
aciatica L., Glycyrrhiza glabra L.) 60 ml each twice a
day, Vacha Ashvaganda [Acorus calamus L., Wilhania
somnifera L. Dunal] powder tea spoon at night with
bee honey, and Mahadalu Anupana [6] with
Chandrakalka [7] 250mg twice a day with bee honey. As
an external treatment, used Narayana oil application (Taila
Abyanga) over upper limbs, lower limbs and head. In
third week, Dashamooladi Decoction [8] and Bhrahmi
Mandukaparnie Maduyashti Decoction 60 ml. each twice
a day and Vacha Ashvaganda powder teaspoon at
night with bee honey given internally and externally used
Shiro Dara with Narayana oil.
During the period of fourth week, used the same
internal treatment as third week and externally treated
with Pizichil using Narayana oil. During the period of
fifth week, internally treated with Danthimoolade
Decoction [9] and Bhrahmi Mandukaparnie Maduyashti
Decoction twice a day, Vacha Ashvaganda Powder at
night and Saraswatha Powder [10] -tea spoon with bee
honey in the morning. Externally patients were treated
with Sashtika shali Pinda sweda.
After the fifth week, patients were treated three days
with Darthri Powder [11] tea spoon at night, which has
mild purgative action. During the last week of treatment
regimen, internally used Mashabalade Decoction [12]
and Bhrahmi Manduka parnie Maduyashti Decoction 60
ml each twice a day, Saraswatha Powder tea spoon in
the morning with bee honey and Vacha Ashvaganda
Powder tea spoon with bee honey at night.
Administered Vasti treatment (Enema) using one ounce
of Narayana oil.
Physiotherapy treatment schedule
Physiotherapy treatment depended on the affected
limbs and the body parts. It consists of a number of
exercises that include stretching, strengthening, and
positioning. Bracing, abduction pillows, knee immobilizers, wheelchair inserts, sitting recommendations and
handling techniques were used in physiotherapy
according to the affected parts of the body.
Duration for physiotherapy was 45 minutes per day.
It continued with three rounds of treatment of 45 days,
each round with two months gap and six months follow
up.
Assessment criteria
The criteria for assessment of treatment was based
on gross motor function classification system (GMFCS)
[13]. Before starting the treatment for the children, they
were assessed by using the assessment criteria of
GMFCS. Then patients were categorized according to
the age and levels of their conditions. GMFCS contains
of five levels. Level 1 is the best (maximum improvement)
and level 5 is the least (minimum improvement). After
completion of the treatment, they were assessed by using
GMFCS, compare pre treatment, and post treatment
levels.
Statistical method
Inter group statistical analysis was performed by
using sample t test of pre-treatment and post-treatment.
Level of significance was set at p < 0.05 in both groups.
Discussion
The efficacy of each therapy was studied by using
GMFCS and results were derived after subjecting to
statistical analysis.
In group A, only five patients were found between
the age from 1-2 years. One patient was in level 5, three
were in level 4 and one was in level 3 at the beginning of
Ayurvedic treatments. After completion of this treatment
regimen, they were found to be in the levels of 1, 2, and
3 (Table 1) Seventeen patients were between the ages of
2 and 4 years. All of them had a significant progress and
came forward (Table 2). Eight patients were found
between the ages of 4 and 6 years. Most of them achieved
progress after the Ayurvedic treatment (Table 3).
In Group B, 10 patients were found between the age
of 1-2 years. They too were in the levels of 3, 4, and 5 at
first, after treatments they acquired some improvement.
Some of them remained in the same level even after
treatment (Table 1). There were only 10 patients between
the age group of 2 and 4 years. Some patients remained
in the same levels even after the treatments (Table 2).
Only 10 patients were found between the age groups of
4 to 6 years. After treatment, they achieved some
progress. However, no one succeed to the level 1 and 2
(Table 3).
57
Original Paper
Table 1: Effects of treatment of patients between 1 and 2 years of age (Group A and B) (n=15)
GMFCS
Levels
No. of patients
before treatment
No. of patients progressed after treatment (GMFCS Levels)
Level 1
Level 2
Level 3
Level 4
Level 5
Total
10
Table 2: Effects of treatment of patients between 2 to 4 years of age (Group A and B) (n=27)
GMFCS
Levels
No. of patients
before treatment
Level 1
Level 2
Level 3
Level 4
Level 5
10
Total
17
10
10
Table 3: Effects of treatment of patients between 4 to 6 years of age (Group A and B) (n=18)
GMFCS
Levels
No. of patients
before treatment
Level 1
Level 2
Level 3
Level 4
Level 5
Total
10
58
Original Paper
Table 4: Intra Group analysis of the overall effect of Ayurvedic Treatment Regimen
and Physiotherapy Trearment regimen
Group
Mean
SD
Remarks
BT
AT
Ayurvedic
(Group A)
4.133333
2.333333
0.62881
0.844182
11.64127
9.44E-13
Significant
Physioth.
( Group B)
4.275862
3.413793
2.067816
1.847645
7.260223
3.32E-08
Significant
P values are significant at p < 0.05
behavioural disorders. Indian Journal of Paediatrics 2005;

72: 865-6.
According to the Table 4, Inter group analysis of pre

treatment and post treatment progress levels of both the
groups revealed significant p value (p < 0.05). The p
value of treated group is 9.44E-13 whereas p value of
physiotherapy group is 3.32 E-08.
4.
US Census Bureau, International Data Base 2004.
5.
Ailapperuma IAS. Vatika Prakaranaya Havest Guli Kalka

Pota, Jinalankara Piriwena, Chandra kalka Anupana. 1915;
221: 2424.
Conclusion
6.

Pota, Jinalankara Piriwena, Chandra Kalka Anupana. 1915;
221: 2425.
7.

Pota, Jinalankara Piriwena, 1915; 220: 2421, 2422.
8.
Ayurvedic Pharmacopia, Department of Ayurveda. 1975;

Part I, Vol I: 98.
9.
Ayurvedic Pharmacopia, Department of Ayurveda. 1975;

Part I, Vol I: 97.
Ayurvedic treatment regimen and Physiothrapy have

the capacity to improve the gross motor functions of
Balaka Pakshaghatha/Cerebral Palsy. However, ayurvedic
treatment regimen is highly effective than that of
Physiotherapy treatment in the management of Balaka
Pakshaghata/Cerebral Palsy. Further studies are proposed
to evaluate the efficacy of combine therapy, i.e. Ayurvedic
treatment regimen and Physiotherapy for the management
of Balaka Pakshaghatha/Crebral Palsy.
Reference
1.
Cerebral palsy hope through research. National Institute

of Neurological Disorders and Stroke (Retrieved on 13 July
2007).
2.
Kumarasinghe A, Charaka Samhita, Department of

Ayurveda,1994; 2nd Edition, Chikitsa Stana/28.
3.
Shankar C, Mandkur N. Symposium on developmental and
10. Ayurvedic Pharmacopia, Department of Ayurveda. 1975;

Part I, Vol I: 127.
Part I, Vol I: 123.
Part I, Vol I: 97.
13. Robert P, Peter R, Stephan W, Dianne R, Ellen W, Barbara
G. Gross motor function classification system for cerebral
palcy. Dev Med Child Neurol 1977; 39: 214-23.
Original Paper
59
Clinical efficacy of Dashamoola Taila Matra Vasti on management of

primary dysmenorrhoea
Kaumadi Karunagoda1, Shilpa Donga2, Lakshmi Priya Dei3
Abstract
Primary dysmenorrhoea is the most common
gynaecological complaint among young women.
Prevalence of dysmenorrhoea is estimated as 45% to
85% among reproductive aged women. It is a condition
which has no any underline pelvic pathology or
anatomical defect. In modern sciences, nonsteroidal anti
inflamatory drugs and oral contraceptive pills are used
as a symptomatic treatment for this condition but they
are having many side effects and they are not curative.
Hence study was carried out to find out a reliable and
longlasting Ayurvedic management for the condition and
to find out the efficacy of Dashamoola Taila Matra Vasti
on primary dysmenorrhoea. The dose of Dashamoola
Taila Matra Vasti was 60 ml and duration was 7 days for
two consecutive cycles. Results were assessed on the
basis of specially prepared grading system for pain. The
results obtained were highly significant. Total effect of
therapy was, 38.89% got complete remission while
marked improvement was there in 50%. In the follow up
period no patient complaint recurrence of symptoms.
The study suggests that Dashamoola Taila Matra Vasti
can be established as a reliable longlasting treatment
for relieving primary dysmenorrhoea.
reproductive aged women [3]. It is a condition which has

no any underline pelvic pathology or anatomical defect
[4]. There is several pathophysiologies and risk factors
have been identified as a etiology of this condition.
Treatment in practice for the condition depends upon
analgesics and oral contraceptive drugs which give several
unwanted effects as well as short term. Several Ayurvedic
oral therapies give significance results on management of
primary dysmenorrhoea without adverse effect, but its
long lasting effect is debatable. Though Uttara Vasti has
proven long lasting effect on this condition it cannot be
implemented to unmarried girls who are the most common
sufferers of Primary dysmenorrhoea [4].
Pain is the main feature of primary dysmenorrhoea,
so it has strong relation with Vata Dosha. Matra Vasti was
taken as a treatment since Vasti has been mentioned as
one of the best therapeutic procedure for alleviation of
vitiated Vata [5]. The present study was carried out as a
very preliminary step to find out a reliable and longlasting
Ayurvedic management for the condition and to find out
the efficacy of Dashamoola Taila Matra Vasti on primary
dysmenorrhoea.
Introduction
Selection of drug
Dysmenorrhoea is a medical condition characterized

by severe uterine pain during menstruation. While many
individuals experience minor pain during menstruation,
dysmenorrhoea is diagnosed when the pain is so severe
as to limit normal activities, or requires medication [1]. It
has been defined as painful menstruation of sufficient
magnitude so as to incapacitate day to day activity [2].
Primary dysmenorrhoea is the most common gynaecological complaint among young reproductive age
women. By 40 high quality studies, prevalence of
dysmenorrhoea is estimated as 45% to 85% among
Kashtartava especially manifesting as primary

dysmenorrhoea is a Vata predominant condition and
selected drugs are also good Vatashamaka drugs as
mentioned in classics. Dashamoola Taila has been
mentioned for the treatment of Udavarta Yonivyapad [6],
which is one of the main disease conditions coming under
primary dysmenorrhoea in Ayurveda.
Contents of Dashamoola [7, 8] are Aegle marmelos
Corr., Premna integrifolia L., Oroxylum indicum Vent.,
Steriospermum suaveolens DC, Gmelina arborea Roxb.,
Desmodium gangeticum DC., Uraria picta Desv.,
Department of Prasutitantra Kaumarabhritya, Institute of Indigenous Medicine, University of Colombo,

2
Department of Stree Roga and Prasutitantra, Institute for Postgraduate Teaching and Research in Ayurveda,
Gujarat Ayurved University, Jamnagar, India.
3
Department of Stree Roga and Prasutitantra, Institute for Postgraduate Teaching and Research in Ayurveda,
Gujarat Ayurved University, Jamnagar, India.
Correspondence: Dr. Kaumadi Karunagoda, Lecturer, Department of Prasutitantra Kaumarabhritya, Institute of
Indigenous Medicine, University of Colombo, Rajagiriya, Sri Lanka. E-mail: kaumadik@gmail.com. Received 20
August and revised version accepted 15 November 2011.
Kaumadi et al. Clinical efficacy of Dashamoola Taila Matra Vasti SLJIM 2011; 01(02): 59-63
60
Original Paper
Solanum indicum L., Solanum surattense Brum and F.,

Tribulus terrestris L.
Ten ingredients of dried Dashamoola are collected
from the pharmacy, identified with the help of organoleptic
and powder microscopic studies. Equal amount of
Dashamoola made in to Yavakuta form is dipped in water
for overnight and next day Kwata is prepared. This Kwata
along with Kalka of Dashamoola is added in Tila Taila
(sesame oil) and oil is prepared as per standard protocol
[9].
Selection of patients
Patients attending the O.P.D. and I.P.D. of Department
of Striroga and Prasutitantra, Institute of Postgraduate
Training and Research in Ayurveda, Gujarat Ayurved
University, Jamnagar complaining of pain in menstruation
and fulfilling the criteria of inclusion were selected for the
present study. An elaborative case taking proforma was
specially designed for the purpose of incorporating all
aspects of the disease on Ayurvedic and modern parlance.
Patients of age group between 15-25 years, coming with
chief complaint of painful menses from more than 3 cycles
with scanty or average amount of menses. Patients below
15 years and above 25 years, patients with chronic illness,
patients with intrauterine contraceptive devices, patients
with menorrhagia or any uterine pathology fibroid,
adenomyosis, endometriosis etc. were excluded from the
study.
Investigations
Routine haematological and urinary examinations
were done before and after teatment. Sonography for
uterine and adnexal study was done for exclude
pathological cases.
left hand below the head. 60 ml of lukewarm Taila was

taken in enema syringe. Rubber catheter oleated with Taila
was attached to enema syringe. After removing the air
from enema syringe, rubber catheter was administered into
the anus of the patient up to the length of 4 inches. The
patient was asked to take deep breath while introducing
the catheter and drug.
Criteria of assessment
The effect of the therapy was assessed considering
to the overall improvement in signs and symptoms. For
this purpose, following categories were maintained.
Severity of pain (multidimensional scoring pattern)
0 Menstruation is not painful and daily activity
unaffected
1 Menstruation is painful and daily activity not affected.
No analgesic required.
2 Menstruation is painful and daily activity affected.
Analgesic drug were needed.
3 Menstruation is painful, she cannot do even her normal
routine work and has to absent from class / office
during menses. Had to take analgesic but poor effect.
Duration of pain
0 no pain in menstruation
1 pain persist less than 12 hours
2 pain continue for 12 -24 hours
3 pain continue more than 24 hours
Criteria for the assessment of overall effect of the
therapies:
Complete remission: 76%-100% relief in the signs and

symptoms were considered as complete remission.
Daily 60 ml of Dashamoola Taila was administered in

morning hours through rectal (Matra Vasti) for 07 days for
two consecutive menstrual cycles starting from mid cycle.
After stopping the administration of the drugs under trial,
patients were advised to report weekly for follow up study,
which was carried out for 2 months.
Marked improvement: 51%-75% relief in the signs and

symptoms were considered as markedly improvement.
Improved: 26%-50 % relief in the signs and symptoms.
Unchanged: Below 25% relief in the signs and

symptoms were considered as unchanged.
Method of administration of Matra Vasti
Investigations : Laboratory investigations Hb%, WBC/

DC, ESR, PCV, were carried out before and after treatment
to rule out any other pathological conditions as well as to
record any specific change by the treatment.
Method of administration
The patient was advised to take light meal, not more

than 3/4th of the usual quantity. Before administration of
Vasti, Abhyanga (application of oil) with Tila Taila was
done on the region of lower back and lower abdomen.
Thereafter, Nadi Sweda was performed.
After this pre preparatory measures (Purvakarma),
the patient was advised to take left lateral position with
left lower extremity straight and right lower extremity flexed
on knee and hip joint. The patient was asked to keep her
Results
Total 20 patients were registered for the study among
them 18 patients had completed the treatment and 02 left
against medical advice. So observations and results drown
from 18 completed patients.
Kaumadi et al. Clinical efficacy of Dashamoola Taila Matra VastiSLJIM 2011; 01(02): 59-63
61
Original Paper
Table 1: Risk factors
Risk factor
No. of patients (n=18)
Early age (<20years)
14
77.77%
Early menarche (12-13years)
11
61.11%
Positive family history
12
66.66%
Lose weight (BMI <20)
33.33%
Null parity
15
83.33%
Smoking
0%
Table 2: Features related to primary dysmenorrhoea

Features
No. of patients
Chronicity (4-6 years)
50%
Onset of pain (1 day before)
11
54.28%
Aggravation of pain (1st day)
17
94.28%
Severity of pain (grade 2)
12
65.71%
Site of pain (Hypogastrium)
18
100%
Duration of pain (12-24hrs.)
10
60%
Table 3: Effect of therapy (% of relief)

Associated symptoms
% of relief
Severity of Pain
75%
Duration of Pain
68.33%
Table 4: Effect of therapy (paired t test)

No of pt.
Mean
B.T.
A.T.
%of relief
S.D.
S.E.
Severity of pain
18
2.23
0.67
70%
0.922
0.22
7.159
<0.001
Duration of pain
18
2.00
0.83
63.89%
0.826
0.19
6.559
<0.001
Table 5: Total effect of therapy

Effect of therapy
No. of pts.
Complete remission (76%-100% relief )
07
38.89%
Marked improvement (51%-75% relief)
02
11.11%
Improved (26%-50% relief)
09
50%
Unchanged (25% -0% relief)
00
0%
62
Original Paper
Discussion
In this study maximum numbers of patients were
suffering form dysmenorrhoea since 4-6 years (Table 2)
among them grade 2 type of severity was found in 65.71%.
The 48.71% of patients were on analgesics/antispasmodics
(Table 3). The data are suggestive of the chronicity of the
problem and also supports the reality that even people
suffering from such common problem visit Ayurvedic clinic
quite late and after taking several other trials.
Majority (54.28%) of them had onset of pain one day
before to the onset of menstruation (Table 3), pain
aggravation on first day on menstruation was found in
94.28% and pain persisted for 12 to 24 hours in 60% (Table
3). These observations show typical characters of primary
dysmenorrhoea [3]. It is a known fact that two days prior
to onset of menstruation, large quantity of progesterone
and estrogen secretes from corpus luteum. This high level
of progesterone induce increases the tone in the isthmus
and upper part of the cervix [4]. An exaggeration of this
could therefore be the basis of the non-coordinating action
of the uterus. Again high level of ovarian hormones stop
FSH (Folicular Stimulating Hormone) and LH (Lutinising
Hormone) secretion causes sudden stoppage in secretion
of progesterone and oestrogen which results to menstruation. Withdrawal of progesterone preceding
menstruation probably causes break down of lysosomes
and the synthesis of various prostaglandins [10]. These
prostaglandins are responsible for pain in menstruation
by myometrial contraction, vasoconstriction and increase
sensitivity of nerve endings for pain [11]. It is the course
of strong pain on starting of menstruation and withdraws
after 24 hours as prostaglandins are of short lifespan [12].
On the basis of site all the patients showed pain in
hypogastrium (Table 3), while in 71.43% complain pain
radiated towards inner and front aspect of the thighs. It
could be because sympathetic nerves, arising from
segments T5 and T6 in the case of motor nerves and from
segment T10 to L1 in the case of sensory nerves, pass
down from the celiac plexus through the intermesentric
plexus, lying retroperitoneally in the front of the abdominal
aorta [13].
Discussing the risk factors of disease, it is similar to
those mentioned for dysmenorrhoea. Early onset of
menarche and early age; below 22 years, which are
mentioned as risk factors for dysmenorrhoea, were found
in 61.11% and 77.77% (Table 2) patients respectively.
The theory postulated behind this finding is that in
this age, pituitary and other endocrine gland do not attain
their maturity till the age of 20 [14]. It can lead to hormonal
imbalance and thus, dysmenorrhoea.
Concept of 'Bija' given as the Nidana of Yonivyapada
in classics was supported by the data obtained, as positive
family history was found in 66.66% (Table 2) of patients. It
suggested that there is a definite relation of a persons
genetic trait and Prakriti with the condition of
dysmenorrhoea. Nulliparity, which is considered as the

risk factor for primary dysmenorrhoea was found in 83.33%
(Table 2) of patients. It is believed that vaginal delivery
removes the stenosis or pin hole of cervical canal and
internal os [1 ] and thus, facilitates the flow of menstrual
blood. It reduces that pain during menstruation, because
the flow of blood through the cervix becomes easier. Lose
weight also a risk factor given as per modern science.
Present study it is noticed 33.33% (Table 2) patients are
below 20 in B.M.I. (Body Mass Index). It supports the
modern findings regarding in this issue [15].
Effect of therapy on primary dysmenorrhoea shows 75%
relief on severity of pain and 68.33% relief in duration and
(Table 4) according to t value highly significant results
(P < 0.001) on both the components (Table 5) evidence
that treatment is effective on both severity as well as
duration of menstrual pain. When considering total effect
of therepy 50% of patients got complete o marked
remission. Not a single show negative respond to the
therapy (Table 6).
In follow up period, most of the patients show
prolong and lasting effect on primary dysmenorrhoea. The
prolongation in recurrence of symptom can be due to the
strong anti-inflammatory and analgesic property of
Dashamoola. It is a proven drug, effective on primary
neurological disorder and improves nerve conduction
velocity [16]. This effect of Dashamoola on nervous
system can be the responsible factor behind its lasting
effect.
Conclusion
Dashamoola Taila Matra Vasti is effective to relieve
primary dysmenorrhoea. Matra Vasti seems to be better
than oral analgesics and oral contraceptive pills used in
dysmenorrhoea, because it is found efficacious in whole
the feature complex related to dysmenorrhoea.
Dashamoola Taila Matra Vasti helps to protect from the
recurrence of dysmenorrhoea. With some further
researches, Dashamoola Taila Matra Vasti can be
established as line of treatment for Primary dysmenorrhoea.
Reference
1.
Dutta DC. The Text Book of Gynaecology, New Central

Book Agency (P) Ltd, Kolkata 2007; 30.
2.
Andrew A. Primary Dysmenorrhoea. American Family

Physician 1999: 06, No. 02, (Retrived on 08/07/2009).
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Howkins & Bourne. Shaws Textbook of Gynaecology, eds

Padubidri VG, Daftary SN. Elsevier India Private Limited,
New Delhi, 2004.
4.
Jeffcoates Principles of Gynaecology, eds. Kumar P,

Malhotra N. Jaypee Brother Medical Publishers (P) Ltd,
New Delhi, 2008; 618.
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Vagbhata. Ashtanga Samgraha, with Hindi Commentry edt.

Kaviraj Atridev Gupta, Krishnadas Aadamy, Varanasi 2002;
(Sutra./19/67).
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Original Paper
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Srikantha Murthy. Vagbhata, Astanga Hrdayam, Krishnadas

Academy, Varanasi. 2001; Vol 3 (A.H./Uttara/39/42).
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Anonymous. The Ayurvedic Pharmacopoeia of India, First

edition, Govt. of India, 2006; Part 1 vol 1.
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Central Council for Research in Ayurveda and Siddha

Database on Medicinal plants used in Ayurveda,
Department of Ayush, Ministry of Health and Family
Welfare, Govt. of India, New Delhi, 2004, Vol 01: 08.
Sharangadhara, Sarngadhara Samhita,eds. Srikanta Murthy
K.R. Chaukambha Orientalia, Varanasi, 2001;115.
10. Rajan R. Postgraduate Reproductive Endocrinology, Jaypee

Brothers Madical Publisher (P) LTD, New Delhi, 2004.
11. Dutta DC. The Text Book of Gynecology, New Central
Book Agency (P) LTD, Kolkata. 2007; 3.
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12. Rajan R. Postgraduate Reproductive Endocrinology, Jaypee

Brothers Madical Publisher (P) LTD, New Delhi, 2004;
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13. Dutta DC. The Text Book of Gynecology, New Central
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Gynaecology, Oxford University Press, New Delhi, 2004.
15. Andersch B, Milsom I. An epidomiologic study of young
women with dysmenorrhea. American Journal of Obstetrics
and Gynecology, 1982; 144: 655-60, www.emedicine.com
(retrieved on 01/10/2008)
16. Chen C, Cho SI, Damokash AI, Chen D, Li G, Wang X.
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64
Original Paper
In vitro evaluation of different aqueous extracts of Senna alata leaves

for antibacterial activity
E Christy Jeyaseelan1, S Tharmila1, A C Thavaranjit1
Abstract
The present study was to enrich the knowledge of
antimicrobial activity of aqueous extracts (cold, hot and
fresh juice) of leaf of Senna alata (L.) Roxb. and to confirm
their effect through qualitative phytochemical analysis.
The preliminary antibacterial assay was performed
against Bacillus subtilis, Staphylococcus aureus,
Escherichia coli, Pseudomonas aeruginosa and Proteus
vulgaris by agar well diffusion method. The Minimum
Inhibitory concentration (MIC) and Minimum Bactericidal
Concentration (MBC) of the cold and hot extracts were
determined by macro broth dilution method. Streptomycin
and sterile distilled water were used as the standard
and control respectively. Qualitative phytochemical
analysis was done to identify the chemical compounds
present in the extracts. The antibacterial activity of the
test extracts differed significantly (P <0.05); the hot and
cold extracts were able to inhibit the growth of all tested
bacteria, while the fresh juice inhibited only B. subtilis
and S. aureus. The cold extract revealed significantly (P
<0.05) higher inhibition on all test bacteria except B.
subtilis, which was highly inhibited by fresh juice. The
MIC of the cold extract ranged from 5 mg/ml to 40 mg/
ml. The lowest value was against P. vulgaris. The MBC
of the cold and hot extracts ranged from 20 mg/ml
to 160 mg/ml. Phytochemical analysis revealed the
presence of glycosides, alkaloids, saponins, cardiac
glycosides, tannins, phlobatannins, flavonoids,
terpenoids and anthraquinones. This study has proved
the feasibility of in vitro control of tested bacteria by
aqueous extracts of S. alata leaves.
Introduction
Senna alata (L.) Roxb (formerly known as Cassia
alata Linn.) (Family-Fabaceae) (Tamil-Vandugolli, SinhalaEt-tora) is a tropical plant, widespread in South East
Asian countries [1]. It is a large shrub with very thick,
finely downy branches; leaves large, sub sessile and
pinnate; flowers are irregular, bisexual, golden yellow in
spiciform pedunculate racemes; pod is long, ligulate with
a broad wing down the middle of each valve [2]. This
plant is traditionally used for the treatment of various
ailments including several infections caused by bacteria,
protozoa, fungi and viruses. The aqueous leaf extracts are
used in the treatment of ringworm and parasitic skin

diseases [3]. In Belgian Congo, the plant is employed as a
remedy for leprosy. In Ghona the leaves are crushed,
mixed with black peppers and applied on dhobys itch,
craw craw and ringworm on the head and skin. In the
Pacific Island and Mauritius the leaves are used for skin
disease [2].
Owoyale et al. (2005) carried out antibacterial and
antifungal screening of different organic solvent extracts
of Senna alata, and they reported that the flavonoid
glycoside is an active ingredient of the extracts [4].
Adedayo et al. (1999) demonstrated the antifungal activity
of methanolic crude extract and partially purified fractions
of flowers of Senna alata against standard and local
fungal isolates [5]. Sule et al. (2010) reported the in vitro
control of fungi; Microsporum canis, Trichophyton
jirrucosum, Trichophyton mentagrophytes and
Epidermophyton jlorrcosum using hot ethanol leaf extract
of Senna alata [6]. The present study is an attempt to
enrich the knowledge of antibacterial activity of different
forms of aqueous extract of leaf of S. alata against some
selected bacteria known to be pathogenic in human. The
phytochemical components were also investigated as a
scientific assessment of the claim of therapeutic potency
of the extracts.

Preparation of plant extracts
The healthy plant leaves were collected from botanical
garden of Department of Botany, University of Jaffna, Sri
Lanka.They were dried in shade. Completely dried leaves
were ground into fine powder using an electric blender.
The powder was used to get cold and hot aqueous
extractions as described below.
a). Cold extract
20 g powder was soaked in 60 ml sterile distilled water
with intermittent shaking for one hour at ambient
temperature. Then the mixture was filtered through doubled
layered muslin cloth and the filtrate was further filtered
through Whatman no 1 filter paper. The filtrate was
completely dried in an oven at 45 C [7].
Department of Botany, Faculty of Science, University of Jaffna, Sri Lanka.
Correspondence: E. Christy Jeyaseelan, Department of Botany, Faculty of Science, University of Jaffna, Sri Lanka.
E-mail: christyjeyaseelan@gmail.com. Received 15 August and revised version accepted 10 November 2011.
Jeyaseelan et al. Antibacterial activity of Senna alata leaves SLJIM 2011;01(02): 64-69
Original Paper
65
b). Hot extract
f). Test for cardiac glycosides
20 g powder was soaked in 60 ml sterile distilled water and

kept in boiling water bath with intermittent shaking for
one hour. Then the mixture was filtered through doubled
layered muslin cloth and the filtrate was further filtered
through Whatman no 1 filter paper. The filtrate was
completely dried in an oven at 45 C [8].
1 ml of concentrated H2SO4 was taken in to a test tube. 5

ml of extract was mixed with 2 ml of glacial CH3CO2H
containing one drop of FeCl3.The above mixture was
carefully added to the 1 ml of concentrated H2SO4. Presence
of cardiac glycosides was detected by the formation of a
brown ring.
c). Fresh extract
g). Test for phlobatannins
20 g fresh healthy leaves were crushed with 20 ml distilled

water using motar and pestle. The crushed material was
filtered through two layered muslin cloth, and the filtrate
was further filtered through Whatman no 1 filter paper.
The filtrate was immediately used for the study [9].
10 ml of extract was boiled with 1% HCl in a boiling tube.

Deposition of a red precipitate indicated the presence of
phlobatannins.
Test microorganisms
Test bacteria, Bacillus subtilis, Staphylococcus aureus,
Escherichia coli, Pseudomonas aeruginosa and Proteus
vulgaris were obtained from a bacterial culture collection,
Department of Botany, University of Jaffna, Sri Lanka.
Phytochemical analysis
The phytochemical analysis of the fresh leaf juice was
carried out to determine the presence of the following
biomolecules using the standard qualitative procedures
as described by Trease and Evans (1989) [10].
a). Test for tannins
1 ml of distilled water and one to two drops of ferric chloride
solution were added to 0.5 ml of extract solution and
observed for brownish green or a blue black coloration.
b). Test for terpenoids
5 ml of extract was mixed with 2 ml of CHCl3 in a test tube.
3 ml of concentrated H2SO4 was carefully added along the
wall of the test tube to form a layer. An interface with a
reddish brown coloration indicated the presence of
terpenoids.
c). Test for steroids
0.5 ml of extract was treated with 0.5 ml of acetic anhydride
and 0.5 ml of chloroform. Then concentrated H2SO4 was
added slowly. Bluish green color was observed for
steroids.
d). Test for saponins
5 ml of extract was shaken vigorously to obtain a stable
persistent froth. The frothing was then mixed with three
drops of olive oil and observed for the formation of an
emulsion, which indicated the presence of saponins.
e). Test for flavonoids
A few drops of 1% NH3 solution was added to the 2 ml of
extract in a test tube. A yellow coloration was observed
for the presence of flavonoids.
h). Test for Alkaloids

1ml of 1% HCl was added to the 3 ml of extract in a test
tube. Then it was treated with a few drops of Meyers
reagent. A creamy white precipitate indicated the presence
of alkaloids.
i). Test for Resins
5 ml of copper solution was added to the 5 ml of extract.
The resulting solution was shaken vigorously and allowed
to separate. A green precipitate indicated the presence
of resin.
j). Test for Glycosides
10 ml of 50% H2SO4 was added to the 1 ml of extract in a
boiling tube. The mixture was heated in a boiling water
bath for 5 min. 10 ml of Fehlings solution (5 ml of each
solution A and B) was added and boiled. A brick red
precipitate indicated the presence of glycosides.
k). Test for Anthraquinones
Extract was mixed well with benzene, and then half of
its own volume of 10% ammonia solution was added.
Presence of a pink, red or violet coloration in the ammonial
phase indicated the anthraquinones.
Determination of antibacterial activity

Agar well diffusion method was used to determine
the antibacterial activity. 20 ml molten nutrient agar media
were mixed with 1 ml of 106 colony forming units /ml) each
test bacterial inoculum and poured into sterile Petri dishes
separately. After complete solidification, 8 mm diameter
wells were made using sterile cork borer.
The cold and hot test extracts were dissolved in
distilled water. The wells were filled with filter sterilized
100 l of (50 mg) cold extract, (50 mg) hot extract, fresh
juice, (50 g) streptomycin and sterile distilled water.
Streptomycin and sterile distilled water were used as
standard and control respectively. The plates were
incubated at 37C for 24 hours and antibacterial activity
was determined by measuring the diameter of clear zone
around the well using Vernier caliper [11]. Each experiment
was repeated three times.
66
Original Paper
Determination of Minimum Inhibitory Concentration

(MIC) and Minimum Bactericidal Concentration (MBC)
The minimum inhibitory concentration was
determined by the macro broth dilution method [12]. The
hot and cold extracts were diluted to 320, 160, 80, 40, 20,
10, 5, 2.5, and 1.25 mg/ml and the streptomycin was diluted
from 5.12 mg/ml to 0.02 mg/ml as two fold dilution in
nutrient broth. The tubes were inoculated with 1.0 ml (0.5
McFarland standards) of test bacteria and incubated at
37C for 24 hours. The MIC was taken as the lowest
concentration of test samples that did not permit any
visible growth. For the determination of MBC, two loops
full of culture were taken from each of the broth tubes that
showed no growth in the MIC tubes and inoculated onto
fresh nutrient agar plates. After 24-hour incubation, the
plates were observed for the growth of bacteria. The concentrations of the extracts that showed no growth were recorded
as the MBC. Each experiment was repeated three times.
Statistical analysis
Results were expressed as mean SD of three
experiments. Statistical significance was determined using
analysis of variance and Tukey test at p = 0.05 using
statistical software SPSS Windows version 13.0.
Results
The qualitative tests for the presence of phytochemicals revealed that the fresh juice of S. alata possess
glycosides, alkaloids, saponins, cardiac glycosides,
tannins, phlobatannins, flavonoids, terpenoids and
anthraquinones. But, the tests for steroids and resins did
not show positive results (Table 1).
The hot and cold extracts were able to inhibit the
growth of all test bacteria, while the fresh juice failed to
inhibit the growth of E. coli, P. vulgaris and P. aeruginosa.
The cold extract showed significantly highest inhibition
on all test bacteria except B. subtilis compared to other
two test extracts and the largest zone of inhibition was
produced against P. vulgaris (23.1 0.2 mm). There was
no significant difference between the inhibitory effects
produced by the cold and hot extracts on S. aureus. The

B. subtilis was highly inhibited by the fresh juice of S.
alata leaf (Table 2).
The antibiotic streptomycin inhibited the growth of
all test bacteria except P. aeruginosa. In most of the cases
the diameter of clear zone produced by the (50 mg/ml)
crude test extracts on test bacteria were found to be larger
than that produced by the (50 g/ 100 l) streptomycin to
the respective bacteria (Table 2).
The minimum inhibitory concentration (MIC) of hot
and cold extracts ranged between 5 mg/ml and 80 mg/ml.
The lowest MIC value, 5 mg/ml was exerted by cold extract
on P. vulgaris. The required MIC value of the cold extract
for E. coli, P. vulgaris and P. aeruginosa were found to be
lower than the hot extract. However, for B. subtilis and S.
aureus, the required MIC values were equal in both hot
and cold extracts. The MIC value of streptomycin was
found between 0.04 mg/ml and 1.28 mg/ml. The minimum
bactericidal concentration (MBC) of test extracts ranged
from 20 mg/ml to 160 mg/ml, and the hot and cold extracts
revealed the lowest MBC against P. vulgaris (Table 3).
Table 1: Phytochemical constituents of fresh leaf juice
of Senna alata
Phytochemicals
Glycosides
Alkaloids
Saponins
Cardiac glycosides
Tannins
Phlobatannins
Resins
Flavonoids
Terpenoids
Steroids
Presence / Absence
+
+
+
+
+
+
+
+
-
Anthraquinones
+ present, - absent
Table 2: Antibacterial activity of different form of aqueous extracts of S.alata leaf

Diameter of inhibition zone (mm)*
Gram positive bacteria
Treatment
Gram negative bacteria
B. subtilis
S. aureus
P. vulgaris
E. coli
P. aeruginosa
Fresh extract
20.60.6a
11.60.8b
Cold extract
14.60.7b
14.30.3a
23.10.2a
21.10.8a
19.00.7a
Hot extract
13.80.4b
14.60.5a
21.70.6b
18.80.7b
10.50.5b
Streptomycin
14.50.6
13.60.7
13.50.5
14.80.4
No activity; * Zone of inhibition includes the diameter of well (8 mm); Values are mean SD, Values with different superscript on the same
column are significantly (P < 0.05) different.
67
Original Paper
Table 3: Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC)
of Senna alata leaf extracts and streptomycin
Test bacteria
MIC (mg/ml)
MBC (mg/ml)
Hot
Cold
Streptomycin
Hot
Cold
Streptomycin
B. subtilis
40
40
0.08
80
80
0.32
E. coli
20
10
0.04
40
40
0.08
P. vulgaris
10
0.64
20
20
1.28
S. aureus
40
40
0.64
80
160
1.28
P. aeruginosa
80
20
1.28
160
80
2.56
Discussion
The present study was undertaken to determine the
feasibility of in vitro control of bacteria by using three
different forms of aqueous extracts of leaf of Senna alata.
In indigenous medicine, these three forms of extracts are
widely used for the treatment of various diseases. The
results revealed that both cold and hot extracts were more
effective than fresh juice, and these two extracts were
able to inhibit the growth of both Gram negative and Gram
positive bacterial species selected for this study.
The activity of the plant extracts against both Gram
positive and Gram negative bacteria is an indication of the
presence of broad spectrum antibacterial compounds [13].
Generally Gram positive bacteria shows higher sensitivity
to plant extracts than Gram negative bacteria [14-17]. This
variation is due to the differences in the cell wall structure
and composition of Gram positive and Gram negative [18].
In this study, even though the cold extract had broad
spectrum of activity, the Gram negative bacteria were highly
inhibited than Gram positive bacteria. This suggests that
there might be specific substances which inhibit the
growth of Gram negative bacteria more.
The lower or absence of bacterial growth inhibition
by the fresh juice of the leaf may be due to the lower
concentration of active ingredients which are toxic to
bacteria. Further study with higher concentration may give
better inhibition. The amount of active ingredients in plant
extracts depend on the climate conditions where the plants
grow. Wandee (2010) reported that the amount of
anthraquinone glycosides in the leaves of S. alata varied
with season. In winter (November-February) and summer
(March-May) plants contain the highest amount of total
antraquinone glycosides (1.24% dry weight). But, the
samples collected in rainy season (June-October) contain
only 0.16% dry weight [1]. In the present study, the sample
was collected from botanical garden where the plant is
irrigated well. Therefore, the amount of active ingredients
may be lower than that grow in wild.
The inhibitory effect of a plant extract resulted from

the activity of phytochemicals was present in the extract.
The type of phytochemical present in an extract depends
on the type of solvent used for the extraction and the
mode of extraction. In this study, the plant material was
extracted in three different methods with water. It can be
clearly seen the variation in the inhibitory effect with the
variation of extraction method (Table 2).
The result of this experiment correlates with a former
study, where the E. coli, S. aureus and P. aeruginosa
were inhibited by aqueous leaf extract of S. alata in agar
well diffusion method. In a previous study done by Okoro
et al. (2010) documented that S. aureus was susceptible to
polyphenol extracts of S. alata, while E. coli appeared to
be resistant to the extracts [19]. But in the present study
both bacteria were inhibited by aqueous extracts. In
another study, hot (soxhlet) aqueous leaf extract of S. alata
failed to inhibit the growth of S. aureus and P. vulgaris,
where the antimicrobial screening was performed by agar
disk diffusion method [20]. The variation in the results
may be due to the variation in the extraction method or
method of antibacterial screening or by both. It was
already reported that agar well diffusion method is more
effective than disc diffusion method for antibacterial
screening as filter paper disc composed of cellulose where
many free hydroxyl groups present on each glucose
residues makes the surface of the disc hydrophilic [21],
and therefore if an extract contains cationic active
constituents with a good antibacterial activity it will not
be expressed in disc diffusion method [22]. In the present
study the hot extract showed comparatively lower activity
than cold extract. Generally, treatment of plant extracts to
high temperature could inactivate volatile compounds, but
could also increase the release of active components and
free radicals [7].
The result for the qualitative phytochemical analysis
also correlates with some previous studies [4,7]. It has
been reported that different phytoconstituents have
68
Original Paper
different degree of solubility in different type of solvents

depending on their polarity [7]. In traditional preparations
water is largely used as the solvent.
5.
Adedayo O, Anderson WA, Moo-Young M, Kolawole DO.

Antifungal properties of some components of Senna alata
flower. Pharmaceutical Biology 1999; 37 (5): 369-74.
The inhibition of tested bacteria by S. alata leaf

extracts confirmed their antibacterial activity and this is
most likely due to the action of different phyto-constituents
present in the extract. Owoyale et al. (2005) reported that
the antimicrobial activity of S. alata is associated with the
presence of phytochemicals such as phenols, tannins,
saponins, alkaloids, steroids, flavonoids and carbohydrates [4]. Flavonoids act as cytoplasmic poisons and
also they have been reported to inhibit the activity of
enzymes [23]. Saponins are surface active agents which
interfere with or alter the permeability of the cell wall.
Therefore, this facilitates the entry of toxic materials or
leakages of vital constituents from the cell. Tannins act
by coagulating the cell wall proteins [24]. Anthraquinones
react irreversibly with amino acids in proteins, often
leading to inactivation of the protein and loss of function.
The alkaloids have ability to intercalate with DNA [6].
6.
Sule WF, Okonko IO, Joseph TA, Ojezele MO, Nwanze

JC, Alli JA, Adewale OG and Ojexele OJ. In vitro antifungal
activity of Senna alata Linn. crude leaf extract. Research
Journal of Biological Sciences 2010; 5 (3): 275-84.
7.
El-Mahmood AM, Doughari JH. Phytochemical screening

and antibacterial evaluation of the leaf and root extracts
of Cassia alata Linn. African Journal of Pharmacy and
Pharmacology 2008; 2 (7): 124-9.
8.
Suleiman MN, Emua SA, Taiga A. Effect of aqueous leaf

extracts on a spot fungus (Fusarium Sp) isolated from
compea. American-Eurasian Journal of Sustainable
Agriculture 2008; 2(3):261-63.
9.
Mohana DC, Raveesha KA. Anti-fungal evaluation of some

plant extracts against some plant pathogenic field and storage
fungi. Journal of Agricultural Technology 2007; 4 (1): 11937.
The standard antibiotic streptomycin showed higher

activity with lower MIC and MBC values compared to the
test extracts. Streptomycin is refined and purified product,
whereas the test extracts are a mixture of varies plant
constituents. Some of these constituents can interfere
within them and this ultimately affects the antibacterial
activity of the extract [7]. Therefore, further study with
bioassay guided fractionation and isolation of pure
compound(s) is necessary to authenticate the effect of
these extracts.
Conclusion
The results of the present study have confirmed the
long history of the use of aqueous leaf extracts of S. alata
in traditional medicine for the treatment of microbial
infections. Even though the hot and cold extracts had
inhibition on all test bacteria, the cold extract showed
comparatively better effect. Therefore, cold aqueous leaf
extract of S. alata can be used for antibacterial treatment
and antibacterial drug screening.
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Somchit MN, Reezal I, Elysha Nur I, Mutalib AR. In vitro

antimicrobial activity of ethanol and water extracts of Cassia
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Owoyale JA, Olatunji GA, Oguntoye SO. Antifungal and

Antibacterial Activities of an Alcoholic Extract of Senna
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70
Original Paper
Selection of the most suitable pot height and harvesting stage for higher
growth, yield and oil quality of Vettiver (Vetiveria zizanioides)
N D N Priyadarshani1 , M K T K Amarasinghe1 , S Subasinghe1 , I R Palihakkara1 ,
H K M S Kumarasinghe1
Abstract
Vetiveria zizanioides (L.) Nash is a valuable
medicinal and aromatic plant used in both indigenous
medicine and perfumery industry. Economically most
important part of the Vettiver is root system. Vettiver roots
are directly used for the medicinal purposes and indirectly
for extraction of essential oils. Low yield and poor quality
roots as well as oil are the problems associated with
Vettiver production. Yield and quality of Vettiver roots
depend on climatic conditions, growing media,
agronomic practices, time of harvesting etc. Objective of
the present study was to select the most promising pot
height and harvesting stage in order to enhance biomass production, oil content and quality of Vettiver. A pot
experiment was conducted at Medicinal Plant Garden,
Faculty of Agriculture, from March 2008 to April 2009. Three
pot heights, namely, 35, 40 and 45 cm with four different
harvesting intervals such as 3, 6, 9 and 12 months after
planting were used for this experiment. Data on number
of tillers, number of leaves, dry weight of roots and shoots
were recorded at 3, 6, 9 and 12 months of planting as
different harvesting stages. Root oil contents, chemical
composition of oils such as Khusimol, -Vetivenene, Vetivone, -Vetivone, Iso-valencinol and fiber content
were also analyzed. Results revealed that, Vettiver
planted in 45 cm pot height showed higher biomass
production. Oil content of Vettiver increased with the
increasing harvesting intervals. Higher oil content
(2.15%) was recorded 12 months after planting.
Subsequently higher oil percentage (2.13%) was
recorded in 9 months after planting. However, there were
no significant differences between oil content of 9 and
12 months after planting. It was also observed in the
present study that the Vettiver harvested at 9 months of
planting had significantly (P< 0.05) high Khusimol
(14.5%), -Vetivone (1.4%) and Iso-valencinol (4.9%)
contents in root oil. Relatively lower fiber contents (36%)
were associated with 9 months after planting compared
to 12 months after planting. Therefore, Vettiver planted in
45 cm pot height and roots harvested at 9 months after
planting could be used as most promising pot height

and harvesting interval in order to enhance bio-mass
production, oil content and quality of Vettiver.
Introduction
Vetiveria zizanioides (L.) Nash (Sinhala Sevendara,
Tamil Vettiver) which belongs to the family Poaceae is
one of the most important medicinal and aromatic plants
widely used in indigenous medicine and perfumery
industry. Vettiver oil is one of the most valuable product
of Vettiver roots. Vettiver oil has 442 extensive applications
in the soap and cosmetic industries, pharmaceutical
companies and as antimicrobial and anti-fungal agent [1].
In Sri Lanka, annual national demand for Vettiver is 41175
Kg (dry basis) and this is valued as 4 million rupees [2].
The root system of Vettiver consists of long fibrous roots
and rootlets. These roots grow more than 2 m in depth
and about 80% of the roots can be found in the first 30-35
cm [3]. Even after the careful harvesting, 40% of the roots
remain in the soil yielding highly damaged roots. One of
the main problems in Vettiver production is poorly
developed low quality roots. These roots produce lower
oil yields as well as low quality oils. Such problems in
Vettiver production could be avoided by adopting proper
agronomic and crop management practices. Therefore, the
present study was carried out to select the most promising
pot height and harvesting stage in order to enhance biomass production, oil content and quality of Vettiver.

A pot experiment was conducted from March 2008 to
April 2009 at Medicinal Plant Garden, Faculty of
Agriculture, University of Ruhuna. Three pot heights,
namely, 35, 40 and 45 cm with four different harvesting
intervals such as 3, 6, 9 and 12 months after planting were
used for this experiment. Three different heights of black
polythene bags were filled using top soil: sand (1:2).
Leaves of tillers were cut down by keeping 3 cm from the
base. Tillers were planted in pots keeping one tiller per
pot. Pots were arranged in a Completely Randomized
Design (CRD) with four replicates. Watering was done at
Department of Crop Science, Faculty of Agriculture, University of Ruhana, Mapalana, Kamburupitiya, Sri
Lanka.
Correspondence: Dr. N. D. N. Priyadarshani, Department of Crop Science, Faculty of Agriculture, University of
Ruhana, Mapalana, Kamburupitiya, Sri Lanka. E-mail: nilukanakandala@gmail.com. Received 15 August and
revised version accepted 12 November 2011.
Priyadarshani et al. Suitable pot height andSLJIM 2011; 01(02): 70-75
Original Paper
two day intervals up to four weeks after planting and
thereafter plants were subjected to rain fed condition. Hand
weeding was practiced at two month intervals.
As non destructive measurements, number of tillers
per bush and number of leaves were taken in 3, 6, 9 and 12
months after planting. Roots were harvested manually and
roots were air dried in the laboratory for three weeks period
to a constant weight. Dry weight of roots and shoots
were taken in Vettiver roots harvested at 3, 6, 9 and 12
months after planting as different harvesting stages. Total
root oil content, chemical composition of oil (Khusimol,
-Vetivenene, -Vetivone, -Vetivone, Iso-valencinol) and
fiber content were determined. Vettiver root samples were
air dried in the laboratory for three weeks period and roots
were cut into 1 cm length of root pieces using a secatier.
Then prepared root samples were used for the oil extraction
and the residue after the oil extraction was used for the
analysis of fiber content of Vettiver roots with the four
replicates from each treatment. Samples were subjected to
AOAC method for determination of crude fiber [4]. Oil
content and chemical compounds in oil were analyzed
71
using Steam Distillation Procedure and Gas

Chromatography Internal Normalization method,
respectively. Data with percentage values were subjected
to angular transformation where necessary and analyzed
using ANOVA (analysis of variance) with Statistical
Analysis System (SAS version 6.12).
Results
Effect of different pot height and harvesting stages on
biomass production of Vettiver
A pot height of 45 cm (T3) showed higher root (dry)
weights 84.5 g, 242 g, 641 g and 777 g respectively at 3, 6,
9 and 12 months after planting. At 9 months after planting
it was more than double the root (dry) weight at 6 months
after planting (Figure 1).
Higher shoot weights (dry) were recorded in 45 cm
pot height (T 3) 192.25 g, 584g, 1572.8 g and 1836.8 g
respectively at 3, 6, 9 and 12 months after planting
(Figure 2).
Figure 1: Changes in dry root weight of Vetiver as affected by different pot heights (cm) at
different harvesting stages (3, 6, 9 and 12 months after planting) (=0.05). T1-35 cm, T240 cm T3-45 cm.
Figure 2: Changes in shoot dry weight of Vetiver as affected by different pot heights (cm)
at different harvesting stages (3, 6, 9 and 12 months after planting) (=0.05). T1-35 cm, T240 cm T3-45 cm.
72
Original Paper
Figure 3: Changes in number of leaves of Vetiver as affected by different pot heights (cm)
at different harvesting stages (3, 6, 9 and 12 months after planting) (=0.05). T1- 35 cm,
T2- 40 cm T3- 45 cm.
Figure 4: Changes in number of tillers of Vetiver as affected by different pot heights (cm)
at different harvesting stages (3, 6, 9 and 12 months after planting) (=0.05). T1- 35 cm,
T2- 40 cm T3- 45 cm.
Pot height had not shown significant differences (P>

0.05) in number of leaves up to 3 months after planting.
However, numbers of leaves were significantly (P<0.05)
affected by pot height after 6 months of planting.
Significantly higher (P<0.05) number of leaves of 215, 471
and 543 were recorded in 45 cm pot height (T3) at 6, 9 and
12 months after planting respectively (Figure 3).
Similarly, pot height had not shown significant
differences (P>0.05) in number of tillers up to 3 months of
planting. However, it was significantly affected by pot
height after 6 months of planting. A significantly higher
(P? 0.05) number of tillers of 36, 67 and 79 was recorded in
45 cm pot height (T3) at 6, 9 and 12 months after planting
respectively (Figure 4).
All the growth and yield parameters (root dry weight,
shoot dry weight, number of leaves and number of tillers)
of Vettiver were higher in 45 cm pot height (T3) compared
to other treatments (pot height of 35 and 40 cm).
Effect of different harvesting stages on oil content and

quality of Vettiver
Oil content of Vettiver increased with the increasing
harvesting intervals. Highest oil content (2.15%) was
observed 12 months after planting (T4). Subsequently
higher oil percentage (2.13%) was recorded in 9 months
after planting (T3) (Figure 5).
Results of chemical compound analysis revealed
that a significantly highest (P<0.05) Khusimol content
(14.5%) was recorded in Vettiver harvested 9 months after
planting (T3). It varied as 10.5%, 7.7 % and 13.5 %
respectively at the 3, 6 and 12 months after planting.
Khusimol content was higher in 3 months old plants
(10.5%) than in the 6 months old plants (7.7%). Production
of Khusimol showed a twofold increase 9 months after
planting (14.5%) compared to the 6 months after planting
(7.7%) (Figure 6).
Original Paper
Figure 5: Oil content (%) of Vetiver at different
harvesting periods. Means with the same letter are not
significantly different at =0.05. T1-3 MAP, T2-6 MAP,
T3-9 MAP, T4-12 MAP.
73
A significantly (P<0.05) higher -Vetivenene (0.8%)

content was recorded 6 months old plants (T2) compared
to 3, 9 and 12 months old plants (Figure 7).
A significantly higher (P<0.05) -Vetivone content
(1.4%) was recorded at the 9 months after planting (T3)
compared to the other harvesting intervals. Production of
-Vetivone increased during the first nine months and
after that it decreased to 0.8%, when it reached to 12 months
after planting (Figure 8).
Figure 8: -Vetivone content (%) of Vetiver oil at different

Figure 6: Khusimol content (%) of Vetiver oil at different

There was an increasing trend in -Vetivone content

(%) with increasing intervals of harvesting. Significantly
high (P<0.05) -Vetivone content (5.2%) was recorded 12
months after planting (T4) (Figure 9).
Figure 7: -Vetivenene content (%) of Vetiver oil at

different harvesting periods. Means with the same letter
are not significantly different at =0.05. T1-3 MAP, T2-6
MAP, T3-9 MAP, T4- 2 MAP.
Figure 9: -Vetivone content (%) of Vetiver oil at different

74
Original Paper
A significantly higher (P<0.05) Iso-valencinol content

(4.9%) was recorded at 9 months after planting (T3)
compared to other harvesting intervals (Figure 10).
Figure 10: Iso-valencinol content (%) of Vetiver oil at

different harvesting periods. Means with the same letter
are not significantly different at =0.05. T1-3 MAP, T2-6
MAP, T3-9 MAP, T4-12 MAP.
Different harvesting intervals showed significant

differences (P<0.05) in fiber content of roots. Fiber content
of roots increased with the increasing harvesting intervals.
A significantly high (P<0.05) root fiber content (44.1%)
was recorded in Vettiver harvested 12 months after planting
(T4) (Figure 11).
Figure 11: Changes in root fiber content of Vetiver as

affected by different harvesting intervals. Means with the
same letter are not significantly different at =0.05. T13 MAP, T2-6 MAP, T3-9 MAP, T4-12 MAP.
45 cm. Yoon (1993) found that, larger bag sizes of 6 13,

7 15 and 8 12 are considered too large for practical
use and there was a decrease in the number of tillers and
top dry weights production from the largest bag to the
smallest bag, which is in agreement to results in this study
[5]. Chomchalow (2001) reported that digging of soil for
root harvesting may be environmentally undesirable, an
alternative means of growing Vettiver could be in polybags and other containers [6]. He further pointed out that,
this would not only mitigate soil erosion concerns but
also increase cost benefit ratio of Vettiver cultivation for
its roots and root oil, as well as optimum utilization of
degraded lands as poly-bag platforms.
It was reported in the present study that oil content
(1.24%) doubled 6 months after planting when compared
to the oil content (0.63%) at 3 months after planting.
Similarly, when considering the oil content between 6 and
9 months after planting it was nearly double at 9 months
after planting. But there was no such increment between 9
and 12 months after planting. The most promising
harvesting time with respect to the root yield was 9 months
after planting. Therefore, it is not economically viable to
keep extra 3 months in the field as it increases the cost of
production.
Maffi (2002) pointed out that the Vettiver roots give
a yield of about 0.3 to 2 % essential oil depending upon
the biotype, cultural practices, age of roots and mode and
duration of distillation [7]. However, in the present study,
the oil yields were 2.13% and 2.15% respectively, 9 and 12
months after planting on a dry weight basis and there
were no significant differences oil contents between 9
and 12 months. Therefore, harvesting interval of 9 months
after planting could be recommended to obtain an
economically viable root and oil yields.
It was also observed in the present study that the
Vettiver harvested at 9 months of planting (T3) had
significantly (P<0.05) high Khusimol, -Vetivone and Isovalencinol contents in root oil. However, during the period
of 9 to 12 months of planting -Vetivone content (5.2%)
of Vettiver increased while Khusimol, -Vetivone and Isovalencinol contents in Vettiver oil decreased.
There were no remarkable changes in temperature,
monthly average rainfall and number of rainy days up to
nine months of planting. However, there were remarkable
reductions in monthly average rainfall and number of rainy
days during the period between harvesting intervals of 9
and 12 months. Present study was conducted under the
rain fed conditions (watering was done at two day intervals
up to four weeks after planting).
Discussion
There was a positive correlation between biomass
productions of Vettiver and pot height. Increase in pot
heights facilitates the downward movement of roots
providing more space. This may be the reason for higher
growth and yield observed in 45 cm than other treatments.
It is not practically feasible to handle pot heights above
Water stress conditions are highly associated with

the secondary metabolites production of Vettiver. These
may be the reasons for such changes in active ingredients.
Maffei (2002) reported that in North India, there is no
definite period for harvesting and the roots are harvested
both for the manufacture of articles and for oil distillation
when plants are 10-12 months old. Chadha (1995) pointed
out that tremendous diversity of oil composition exists
Original Paper
with respect to pattern of growth, orientation and thickness
of roots, as well as for occurrence of secondary roots and
harvesting time [8]. Aggarwal et al. (1998) demonstrated
that the age, quality and stage of root harvest, and
processing for distillation are vital components for essential
oil distillation [9].
High fiber content reduces the yield and quality of
roots as well as oil. Therefore, it is necessary to select
best possible harvesting interval with lower fiber content
for yield and quality improvement of the Vettiver oil. Anon
(1976) reported that Vettiver has a high content of
hemicelluloses and its cellulose content is 45.8% (Dry
Weight basis) [10]. He also revealed that Vettiver
containing short fiber and pulp has to be used in admixture
with 30-40% of a long-fibered pulp. Though the high fiber
content of Vettiver is important for the paper industry it is
not a good feature in oil distillation as it creates practical
difficulties in processing, oil extraction as well as loses
the essential ingredients in oil.
References
1.
Singh G, Singh BS, Kumar BRV. Antimicrobial activity of

essential oils against keratinophilic fungi. Indian Drugs
1978; 16(2): 43-5.
2.
Abeywardana N, Hettiarachchi LJK. Statistics on the

National Demand for Medicinal Plants, Sri Lanka
Conservation and Sustainable Use of Medicinal Plants
Project 2001; 76-9.
3.
Peyron L. Vettiver in Perfumery, Quintessenza. Taylor

and Francis Publishers, London 1989; 4-14.
4.
American Association of Cereal Chemists (AACC) Dietary

Fiber Technical Committee. The definition of dietary fiber.
Cereal Foods World 2001; 46-112.
5.
Yoon PK. A look-see at vettiver grass in Malaysia, Part 2:

establishment and management of quality vettiver
hedgerows. Vetiver Information Network 1993; 53.
6.
Chomchalow N. The utilization of vettiver as medicinal

and aromatic plants with special reference to Thailand.
PRVN Tech. Bull. No. 2001/1, Office of the Royal
Development Projects Board (ORDPB), Bangkok 2001;
26-9.
7.
Maffei M. Vetiveria (the genus Vetiveria). Taylor and

Francis Publishers, London 2002; 191.
8.
Chadha YR. The Wealth of India a dictionary of Indian

raw materials and industrial products. Raw materials vol.
x-: sp-w, Publications and Information Directorate, CSIR,
New Delhi 1995; 451-6.
9.
Aggarwal KK, Singh A, Kahol AP. Parameters of vettiver

oil distillation. Herbs, Spices and Medicinal Plants 1998;
55-61.
Conclusion
Pot height of 45 cm and Vettiver harvested at 9
months of planting could be used as most appropriate pot
height and harvesting interval in order to enhance biomass production, oil content and quality of Vettiver. Period
of harvesting highly depends on the soil and climatic
conditions, agronomic practices adopted and purpose of
harvesting.
Therefore, further research has to be carried out to
select proper harvesting time in relation to the soil types
and climatic condition of the different regions to obtain
maximum yield in good quality.
Acknowledgement
Department of Ayurveda, Ministry of Indigenous
Medicine is greatly acknowledged for the funds provided
for this research project.
75
10. Anon. The Wealth of India raw materials. Council for

Scientific and Industrial Research, New Delhi, India 1976;
451-7.
76
Original Paper
Anti hyperlipidemic effect of Vara Asanadi Kwatha against high fat diet
induced hyperlipidemic rats
Anju P Ramachandran1, M Shyam Prasad1, Vijay Kumar2, B K Ashok3, B Ravishankar4,
H M Chandola5
Abstract
Changing life style and diet patterns along with
significant role played by genetics made Hyperlipidemia/
Dyslipidemia as one of the most common metabolic
aberration of lipids among people all over the world. An
extra cavernous research study on classical Ayurvedic
formulations which are not in the limelight of routine
clinical practice is essential to explore the most effective
and target oriented anti hyperlipidemic drugs. Objective
of this study was to evaluate anti-hyperlipidemic activity
of Vara asanadi kwatha against high fat diet induced
hyperlipidemic Wistar strain albino rats. Wistar strain
albino rats of either sex weighing 180 25 g six animals
were selected and housed with each cage containing 6
animals. Test drug treated animals were managed with
Vara Asanadi Kwatha at a dose of 8 ml/kg in which the
efficacy of medicine has been assessed on various
serum biochemical parameters, histopathological
sections and weights of liver, kidney and heart. One group
kept as cholesterol control and the remaining as water
control. Findings are in favour of mild anti hyperlipidemic
and significant hepato-protective and nephro-protective
activities of the test formulation. Vara Asanadi Kwatha is
a mild anti-hyperlipidemic and potent hepato-protective
as well as renoprotective drug.
form of decoction, which claims to be effective in the

management of overweight and obesity [3]. An
experimental evaluation of the drug on its antihyperlipidemic action will certainly give thoughts
regarding the efficacy of Vara Asanadi Kwatha in
counteracting the ill effects of dyslipidemia. With this
judgment the present experimental study was carried out
to screen anti-hyperlipidemic potential of VAK in
experimental animals.

Test formulation:
The ingredient wise composition of Vara Asanadi
Kwatha has been provided in Table 1.
Each raw constituent of VAK was subjected to
pharmacognostical identification and was certified as
genuine and of good quality in the Department of
Pharmacognosy, Institute of Post Graduate Teaching and
Research in Ayurveda (IPGT and RA), Gujarat Ayurved
University, Jamnagar. The test drug was prepared by
adding one part of the crushed raw drug to sixteen parts
of water, boiled and reduced to half. Thin sheets of Iron
was added during the boiling period of kwatha and later
removed while filtering. The prepared drug was procured
from Ayurveda Pharmacy, Kannur, Kerala.
Introduction
Lipid and lipo-protein abnormalities have become
enormously common in the general populace. The
metabolic aberrations of lipids are linked as risk factor
with numerous numbers of serious systemic illnesses
including cardio vascular disorders and metabolic
syndrome [1]. Obesity and hyperlipidemia often exist
together clinically and share much in common from the
etio-pathology to the complications [2]. Vara Asanadi
Kwatha [VAK] is a classical Ayurvedic formulation in the
Animals:
Wistar strain albino rats of either sex weighing 180
25g were obtained from animal house attached to
Pharmacology Laboratory of IPGT and RA Gujarat
Ayurved University, Jamnagar. Six animals were housed
in each cage made up of poly-propylene with stainless
steel top grill. The dry wheat (post hulled) waste was used
as bedding material and was changed every morning. The
animals were exposed to 12 hour light and 12 hour dark
Department of Kayachikitsa, Institute of Postgraduate Teaching and Research in Ayurveda (IPGTR&A), Gujarat
Ayurved University, Jamnagar, India.
2
PGT-SFC, Gujarat Ayurved University, Jamnagar, India.
3
Pharmacology Laboratory, IPGTR&A, Ayurved University, Jamnagar, India.
4
SDM College of Ayurveda, Laxminarayana Nagar, Kuthpady, Udupi, Karnataka, India
5
Department of Kayachikitsa, IPGTR&A, Gujarat Ayurved University, Jamnagar, India.
Correspondence: Dr. Anju. P. Ramacahndran, w/o Dr. Shyamprasad. M. Mahalakshmipuram, Thekkil P.O Pin671541, Kasargod (dist) Kerala, India E-mail: anjushyamprasad@gmail.com. Received 24 August and revised
version accepted 10 November 2011.
Anju Ramachandran et al. Anti hyperlipidemic effectSLJIM 2011; 01(02): 76-82
Original Paper
77
Table 1: Composition of Vara Asanadi Kwatha

Plant and part used
Latin name
Quantity
Haritaki
Terminalia chebula
1 part each
Bibhitaki
Terminalia bellerica
Amalaki
Emblica officinalis
Asana
Pterocarpus marsupium
1 Part
Citraka
Plumbago zeylanica
1 Part
Haridra
Curcuma longa
1Part
Triphala
Lohaptra
(Fe2O3)
cycle with the relative humidity of 50 to 70% and the

ambient temperature during the period of experimentation
was 22 03C. Animals were fed with Amrut brand rat
pellet feed supplied by Pranav Agro Mills Pvt. Limited.
For their drinking purpose tap water ad libitum was
used. The experiment was carried out after obtaining the
permission from institutional animal ethics committee.
(Approval number; IAEC (Institutional Animal Ethics
Committee) 06/09-11/PhD/05).
Dose fixation and schedule:
The human dose of Vara Asanadi Kwatha is 45ml
twice a day (90 ml per day) [4]. The suitable dose for rats
was calculated by referring to table of Paget and Barnes
(1964) [5] and the dose obtained thus was 8 ml/kg rat. The
test formulation was administered with the help of oral
catheter attached to a disposable syringe.
Anti-hyperlipidemic activity:
The effect of test formulation on diet induced
hyperlipidemia was carried out as per previous study [6].
The selected animals were divided into three groups of six
animals each. First group was kept as normal control (NC)
which received only tap water.
To second group hyperlipidemic diet was
administered and served as cholesterol control (CC) group.
Third group received hyperlipidemic diet and Vara Asanadi
Kwatha (VAK). Test drug was administered at morning
hour and hyperlipidemic diet (to second and third group)
was administered at evening hours for 20 consecutive
days. The hyperlipidemic diet includes hydrogenated
vegetable oil (Vanaspati Ghee - Raag brand, Batch No.
BA 76, Adani Wilmar Ltd., Gujarat) and cholesterol extra
pure powder (Batch No. 14036 Suvidhnath Laboratories,
Baroda) made in to 20% suspension in coconut oil
(Parachute coconut oil, Batch No. GSW002, PondaGoa.).The suspension was administered at the dose of 0.5
ml/100 g rat. On the 21st day after overnight fasting, the
Thin sheets added on boiling

kwatha (Decoction) and
removed afterwards
animals were weighed again and blood was collected from

retro-orbital plexus under ether anaesthesia. From
separated serum; biochemical parameters like glucose [7],
serum total cholesterol [8], serum triglyceride [9], and
serum high density lipoprotein cholesterol (HDL-C) [10],
Serum low density lipoprotein cholesterol+ very low
density lipoprotein cholesterol (LDL-C + VLDL-C) were
estimated. Serum (LDL+VLDL) was calculated by
subtracting HDL cholesterol value from total cholesterol
instead using both values separately, as in rats whose
serum cholesterol is <100 mg/dl Friedewald formula
overestimates LDL levels [11]. Further blood urea [12],
serum creatinine [(13], serum glutamic oxaloacetic
transaminase (S.GOT), serum glutamic pyruvic
transaminase (S.GPT) [14], alkaline phosphatase [15], total
bilirubin [16], direct bilirubin [17] and serum uric acid[18]
were also estimated as per standard procedure. Further,
all the rats were sacrificed by overdose of ether anesthesia
and from the sacrificed animals liver, kidney, heart and
aorta were excised out. The liver, kidney and heart were
weighed and fixed in 10% buffered neutral formalin solution.
After fixation, tissues were embedded in paraffin and serial
sections were cut and each section was stained with
hematoxylin and eosin [19]. The slides were viewed
under trinocular research microscope (Germany) at various
magnifications to note down the changes in the
microscopic features of the tissues studied.
Statistical analysis:
The results were presented as mean SEM for six
rats in each group. Statistical comparisons were performed
by unpaired students t test by using Sigma stat software
(version 3.1) for all the treated groups with the level of
significance set at P<0.05.
Results
Data related to effect of VAK on body weight of
albino rats have been provided in Table 2.
78
Original Paper
Table 2: Effect on body weight
Treatment
Body weight (g)

Initial body
weight (g)
Final body
weight (g)
Actual change in
body weight (g)
% change in comparison
to control
NC(n=6)
203.00 6.38
218.00 6.97**
15.00 3.49
--
CC(n=6)
174.67 8.88
196.00 11.41*
21.33 5.74
42.20
VAK(n=6)
177.00 3.96
217.33 7.04**
40.33 6.56
89.08
Data: Mean
SEM (standard error of mean), - Increase, *P<0.05, **P<0.01,
(Compared with initial body weight, paired t test)
In normal control rats a progressive gain in body weight was occurred in comparison to its initial values. In contrast
to this, significant increase in body weight was occurred in cholesterol control group in comparison to both initial values.
In VAK treated group also significant increase in body weight was occurred in comparison to its initial value. Marginal
increase of relative weight of liver and heart was found in cholesterol control group in comparison to normal control
group which is found to be statistically non-significant (Table 3).
Table 3: Effect on weight of important organs

Treatment
Weight of liver
(mg/100g)
Weight of heart
(mg/100g)
Weight of kidney
(mg/100g)
NC (n=6)
3061.83 102.96
335.13 6.26
605.00 08.04
CC(n=6)
3356.85 185.82
337.39 14.58
642.19 10.32*
VAK(n=6)
2934.58 43.99
330.69 6.93
591.58 10.42
Data: Mean SEM

*P<0.001(Compared with normal control group, unpaired t test)
P<0.01(Compared with cholesterol control group, unpaired t test)
Treatment with VAK attenuated weight of these organs in non-significant manner. Further, cholesterol control
group significantly increased the kidney weight and treatment with VAK significantly attenuated it. The data related to
the effect of VAK on serum biochemical parameters were provided in Table 4.
Feeding of cholesterol diet led to significant increase in serum glucose in comparison to normal control group and
treatment with VAK non-significantly attenuated it. Further blood urea, serum creatinine and serum lipid profiles were
significantly increased by feeding with hyperlipidemic diet. These parameters were also non-significantly attenuated by
administration of VAK. S.GOT, Aspartate transaminase (AST) and alkaline phosphatase activities were significantly
enhanced by feeding with hyperlipidemic diet in rats. VAK significantly attenuated activity of these enzymes in comparison
to cholesterol control group. Further total bilirubin and serum uric acid levels were also elevated by feeding of hyperlipidemic
diet and VAK significantly attenuated them.
Histopathological sections from control group shows normal cytoarchitecture of liver, kidney and heart (Fig. 1A, 2A
and 3A). In contrast, hyperlipidemic diet produced perivascular cell infiltration and micro fatty changes in liver, cell
infiltration and fatty changes in kidney and cell infiltration and fatty changes in majority of sections of heart (Fig. 1B, 2B
and 3B). Simultaneous treatment with VAK significantly attenuated cholesterol induced pathological changes in all the
three organs (Fig. 1C, 2C and 3C).
79
Original Paper
Table 4: Effect of on various serum bio-chemical parameters

Parameters
NC
CC
% change in
comparison to NC
VAK
% change in
comparison to CC
Blood sugar (mg/dL)
87.50
4.33
111.67
3.34**
27.62
101.67
7.44
08.95
Cholesterol (mg/dL)
58.00
2.87
87.33
2.40***
50.56
80.50
4.93
07.82
Triglyceride (mg/dL)
81.16
2.54
164.00
16.54***
102.06
134.16
15.07
18.19
LDL+VLDL (mg/dL)
27.50
2.51
42.80
4.42*
55.63
42.73
4.38
--
HDL (mg/dL)
30.50
1.25
39.50
3.08*
29.50
38.00
3.58
03.79
Apolipoprotein B
19.83
0.30
14.16
0.94***
28.59
17.16
0.87
21.18
Blood urea (mg/dL)
75.33
3.48
42.33
1.05***
43.80
44.80
4.56
06.37
Serum creatinine (mg/dL)
0.48
0.03
0.68
0.03***
41.66
0.60
0.03
11.76
S.GOT (IU/L)
152.50
5.29
236.00
25.73**
54.75
144.66
11.22
38.70
S.GPT (IU/L)
46.67
3.31
73.00
4.81**
56.41
63.66
2.01
12.79
Alkaline phosphatase
(IU/L)
211.16
9.81
526.33
22.64***
149.25
402.50
28.00
23.52
Bilirubin total (mg/dL)
0.35
0.02
0.55
0.06*
57.14
0.35
0.05
36.36
Bilirubin D (mg/dL)
0.15
0.20
33.33
0.15
25.00
0.02
0.02
0.90
0.10
1.35
0.15*
Uric acid (mg/dL)
Data: Mean
0.02
50.00
0.80
0.11
40.74
SEM, - Increase; - Decrease,
*P<0.05, **P<0.01, ***P<0.001(Compared with normal control group, unpaired t test)
P<0.05,
P<0.01 (Compared with cholesterol control group, unpaired t test)
80
Original Paper
Figure 1A. NC-liver showing normal

cytoarchitecture, Hc-hepatocytes
Kc-Kupffer cell, S-sinusoid.
Figure 1B. CC-liver showing micro

and macro (Fc) fatty changes, (CI)
cell infiltration.
Figure 1C. VAK liver showing

almost normal cytoarchitecture.
Figure 2A. NC-kidney showing

normal cytoarchitecture Gglomerulus; Ct-convoluted tubule.
Figure 2B. CC-kidney showing

micro (Fc) fatty changes, and (CI)
cell infiltration.
Figure 2C. VAK kidney showing

almost normal cytoarchitecture.
Figure 3A. NC-heart showing normal

cytoarchitecture, Mc-myocardium.
Figure 3B. CC-heart showing

CI - cell infiltration.
Figure 3C. VAK heart showing

normal cytoarchitecture.
Discussion
Elevated levels of different types of lipids have been
implicated in the production of atherosclerosis. In this
stipulation the blood vessel wall thickens due to
cholesterol deposition ensuing to inflammatory reaction.
This ultimately leads to loss of elasticity of affected
vessel wall and becomes the major pathology involved
in the occurrence of a number of serious systemic
disorders such as cardio vascular diseases, cerebrovascular accidents, peripheral arterial disease which
account as the significant culprit for mortality/disability
in both developed and developing countries. Even
after the prescription of dietetic, lifestyle and therapeutic

interventions the incidence and prevalence of lipid
abnormalities and resultant fatal complications are hiking
up. Hence there is huge scope for the introduction of
effective hypolipidemic and anti-hyperlipidemic drugs
in to existing therapeutic armamentarium.
In the current experimental work, in comparison to
cholesterol control group, the VAK treated animals
exhibited moderate level of decrease in S.cholesterol,
S.triglycerides and HDL-C, but the variations were
statistically non significant.
Original Paper
81
Statistically significant changes were attained in the

values of SGOT, SGPT, alkaline phosphatase, total bilirubin
and uric acid revealing the high hepatoprotective and
nephroprotective properties of the test formulation. SGOT
determination is of immense value in the assessment of
coronary artery diseases and myocardial infarction.
blood glucose and renal lesions. It had been demonstrated

to reduce smooth muscle cell proliferation and endothelial
dysfunction [26]. As per recent research studies Curcumin
has been reported to have the nephroprotective effect to
improve creatinine and urea clearance and also can protect
the chronic renal allograft nephropathy [27].
Elevated serum enzyme activity associated with

cardiac disorder is assumed to reflect activity of enzyme
released from the injured cardiac tissue too [20]. The
significant change attained in the value of SGOT may also
have noteworthy role in the cardio-protective activity of
the trial drug. The observations attained in bio chemical
parameters are in line with the histopathological findings
of this study. The normal cytoarchitecture and absence of
cholesterol induced pathological changes in the
histopathological sections of liver, heart and kidney shows
the efficacy and capability of VAK in the management of
dyslipidemia induced complications.
Furthermore the hypolipidemic and hepatoprotective

activities of Pterocarpus marsupium (Asana) are also well
established by several studies [28,29]. Most of the
Ayurvedic drugs such as E. officinalis and C. longa are
stronger and efficient anti-oxidants; which may be helpful
in preventing lipid peroxidation [30].
Vara is well known as triphala (combination of

Terminalia chebula, Terminalia bellerica and Emblica
officinalis) in Ayurveda is the foremost ingredient of VAK
and Ayurvedic science has identified its benefits in
obesity, diabetes mellitus and hepatic disorders. There
are many reports with regard to pharmacological effects
of triphala, including its anti-hypercholesterolemic, antioxidant and hepatoprotective properties [21,22]. Emblica
officinalis (Amalaki) given in a ration of rabbit at 1g/kg
has found to have anti-hypercholesterol activity. In one
of the study; T. arjuna, T. bellerica and T. chebula was
fed to rabbits on cholesterol rich diet inducing atherosclerosis which showed that T.chebula as the most potent
hypolipidemic agent among the three drugs and induced
partial inhibition of rabbit atheroma as seen from plasma
and tissue lipid content and the lesions of aorta. Haritaki
(T. chebula) is also well known for its anti-hepatotoxic
activities. Hepatoprotective activity of T.bellerica is also
been reported as the alcoholic extract of fruit of T. bellerica
in a dose of 30 mg/kg given I/V to dogs showed significant
bile stimulant activity and increased solids in bile secretion.
Further numerous studies have been conducted on the
anti-hyperlipidemic acivity of Citraka (Plumbago
zeylanica), which is the one of the ingredient of VAK.
In the study conducted on hyperlipidemic rabbits
Plumbagin; the active constituent of Plumbago zeylanica
reduced serum cholesterol by 53-86% and elevated
decreased HDL cholesterol significantly [23].
Curcuma longa (Haridra) is an established hepato
protective drug and is been used widely in the management
of jaundice and hepatic disorders [24]. C. longa prevents
the formation of fatty liver by the modulation of
expressions of enzymes that are important to fat metabolism
[25].
In a usual mutant obesity, Curcuma longa had
significantly reduced cholesterol and triglyceride
concentration, while increasing HDL cholesterol. Advance
evidences indicate that it diminishes the oxidation of LDL,
Thus multiple constituents of VAK are reported to

have antihyperlipidemic, anti-hepatotoxic and hepatoprotective activities. The ingredient such as Curcuma
longa is having nephroprotective properties also and the
same is reflected in present study. The non-significant
changes obtained in the most of the values of lipid profile
and blood sugar cannot be interpreted negatively, as the
results are definitely pointing towards the direction of
reduction. The weak action obtained in terms of these
parameters may be because of the fact that the drug is
administered in the form of decoction. Otherwise most of
the individual components of the Vara Asanadi Kwatha
are proven anti-hyperlipidemic drugs when used in single
or in combinations. The alcoholic extract of the same drugs
may show more potent and significant anti-hyperlipidemic
activity as reported by the various studies in this regard.
Conclusion
From the present study it can be concluded that
Vara Asanadi Kwatha is having mild anti-hyperlipidemic
and remarkable hepato-protective and nephroprotective
activities. Exclusive experimental works on hepatoprotective and nephroprotective properties of Vara
Asanadi Kwatha may reveal hidden and highly informative
facts regarding this wonderful classical formulation.
Acknowledgement
The authors wish to thank Dr Sulakshan Chavan,
Miss Hetal Aghera and staff of pharmacology laboratory,
IPGT and RA, and Jamnagar for their technical support.
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Short Communication
83
Antibacterial properties of Accmus mouth wash

S Tharmila1, T Thileepan2, A C Thavaranjit1, R Srikaran3
Abstract
Antimicrobial herbs can be used individually or in
combination to prepare mouth wash which is healthier
and safer than the synthetic ones. In this study a new
Accmus herbal mouth wash was prepared and its
antibacterial properties were evaluated. Alcoholic, boiled
alcoholic and aqueous extracts of Accmus mouth wash
were prepared from the bark of Acacia arabica, Acacia
speciosa and root of Calamus rotang in combination by
tincture and hot extract methods respectively. Alcohol
content and pH were also determined. Antibacterial
properties of the above extracts were also studied against
Staphylococcus aureus, Bacillus sp of Gram (+)ve and
Pseudomonas aeruginosa, Klebsiella sp of Gram (-) ve
in vitro by using agar well diffusion method. This study
showed that the alcohol content and pH of mouth wash
preparations were in acceptable levels. Aqueous extract
exhibited better antibacterial activity compared with
alcoholic extract and had maximum sensitivity towards
Bacillus sp and low towards Klebsiella sp. Staphylococcus aureus was only inhibited by all preparations of
mouth wash. So the hot extraction method was efficient
than the alcoholic extraction and this could be
recommended with antibacterial properties rather than
the alcoholic extract of mouth wash. Further study is
needed for further purification and characterization of
active constituents from various solvent extracts of mouth
wash against oral diseases.
Introduction
Mouth wash or mouth rinse is a product used to
enhance oral hygiene. Commercial brands of mouth wash
contain synthetic and semisynthetic chemical substances
such as thymol, methyl salicylate, menthol, chlorhexidine
gluconate, methylparaben, hydrogen peroxide etc [1] and
also include water and sweetness such as sorbitol, sodium
saccharin [2]. Sometimes a significant amount of alcohol
is added as the carrier for the flavour. Sodium benzoate is
a common preservative in commercial mouth washes [3].
The risk of acquiring cancer rises almost five times for
users of alcohol containing mouth wash who neither
smoke nor drink [4]. Mouth washes containing
cetylpyridinium chloride are also associated with loss of
taste sensation and brown discoloration of teeth [4]. To
overcome such harmful effect natural mouth washes are

available in markets and are produced from plant based
healthy ingredients such as organic aloe vera, peppermint,
clove bud essential oils, perilla seed extract etc. The
present study is to prepare a new Accmus mouth wash
from the bark of Acacia arabica, bark of Acacia speciosa
and root of Calamus rotang. Acacia arabica (KaruvelT) is a tree, becomes under family leguminosae. Its bark
has medicinal properties, mainly used in oral diseases.
Hence, it has 24-42% of tannin. Acacia speciosa
(Kadduvakai T) becomes under family mimosaceae.
Its bark decoction is being used in orodental diseases for
gargle. Powder of root bark is used for bleeding. Calamus
rotang is a climber one and it is classified under family
palmae. In traditional medicine the root of Calamus rotang
has been used against many oral diseases such as gum
bleeding and aphthous ulcer in form of decoction for
gargling [5,6]. The objective of this study is to prepare a
natural new Accmus mouth wash and test its
antibacterial activity against Gram (+) ve and Gram (-)ve
bacteria.
Collection of plant materials
The plant Acacia speciosa was collected by the Unit
of Siddha Medicine, University of Jaffna, Sri Lanka and it
was identified based on herbarium records in the
Department of Botany, University of Jaffna and other
relevant materials [7,8]. And healthy bark was obtained,
washed under running tap water, dried in sun shade for
three weeks. Then ground into fine powder. Bark of Acacia
arabica and root of Calamus rotang were also collected
from local market and their characters were compared with
herbarium records [7,8]. The above parts were washed
under running tap water, dried in sun shade for five days
and then ground into fine powder, by using electric
blender. The powder was stored in air tight dark bottles at
room temperature.
Preparation of mouth wash
Accmus mouth wash was prepared by two
methods.
Department of Botany, Faculty of Science, University of Jaffna.

Unit of Siddha Medicine, University of Jaffna.
3
Department of Chemistry, Faculty of Science, University of Jaffna.
2
Correspondence: Miss. S. Tharmila, Assist. Lecturer, Department of Botany, Faculty of Science, University of Jaffna.
E-mail: Tharmila9@gmail.com. Received 15 August and revised version accepted 10 November 2011.
Tharmila et al. Antibacterial properties of Accmus... SLJIM; 01:(02) 83-85
84
Short Communication
Tincture method
25 g of each of the above herbal powder was mixed
and mixture was soaked in 93.75 ml of 25% ethanol and
281.25 ml distilled water for two weeks under direct sun
light with occasional shaking. The mixture was filtered
through double layered muslin cloth and the filtrate (355
ml) was collected into a clean dried dark bottle.
Half of the above volume of the filtrate was boiled at
85C for 30 minutes and poured into a clean dried dark
bottle as boiled alcoholic extract [9].
Hot extract method
25 g of each of the above herbal powder mixture was
mixed with 250 ml distilled water in a sterile beaker. It was
heated at 50C on hot plate for 6 hours continuously till
the final volume of extracts reached as 150 ml. Then
extracts were filtered through double layered muslin cloth
and the filtrate was concentrated by heating. It was kept
at 4C until used for assay [10].
Determination of pH was determined by pH meter.
Determination of alcohol content
Alcohol content of mouth wash was determined by
ebuliometer. Durability of mouth wash also noted based
on its characters such as color change, (odour) smell
formation, turbidity and change in viscosity.
Antibacterial assay
Culture preparation.
The bacterial isolates of Staphylococcus aureus,

Bacillus sp from Gram positives and Gram negative
Pseudomonas aeruginosa, Klebsiella sp were obtained
from bacterial culture collection, Department of Botany,
University of Jaffna for this study. Test organisms were
stored on nutrient agar slants at 4C and these were sub
cultured before 24 hours of the experiment and incubated
at 37C. After the incubation a loop full of young bacterial
inoculum was transferred into the 10 ml of sterile saline
water (0.85%) in an aseptic condition. Inoculum
concentration was estimated by haemocytometer and the
number of cells per ml was adjusted to 106 cells by using
tenfold dilution [11].
Determination of antibacterial activity
Nutrient agar medium was autoclaved and cooled to
40C. The antibacterial assay was performed by agar well
diffusion method [12]. 1 ml of test culture (106 CFU/ml)
was inoculated into a sterile petridish with 20 ml sterile
nutrient agar and mixed well and allowed to solidify. Then
wells were made by using sterile cork borer (8 mm in
diameter) on the surfaces of agar plates and were filled
with 100 l of each extracts using sterile Pasteur pipette.
100 l of commercially available Chlorhexidine
digluconate mouth wash was used as standard and
alcohol and water were used as control. Then plates were
incubated at 37C for 24-48 hours. Antibacterial activity
was determined by measuring the diameter of the clear
zone around the well. The above experiment was repeated
five times and the mean diameter of the zone of inhibition
was calculated.
Results and Discussion

Table 1: Antibacterial activity of mouth wash extracts on test bacteria
Mouthwash extracts
Mean zone of inhibition (mm)

S. aureus
Bacillus sp
P. aeruginosa
Klebsiella sp
(+)ve
(+)ve
(-)ve
(-)ve
Alcoholic extract of mouth

wash (Tincture)
12
Alcoholic extract of mouthwash

after boiling (Tincture)
10
Aqueous extract of mouthwash
15
16
12
10
Chlorhexidine
digluconate (Standard)
16
20
15
13
Zone of inhibition includes the diameter of the well (8mm in diameter). (-) No activity.
Short Communication
85
Table 2: pH and alcohol content of mouth wash extracts

Mouth wash extracts
pH
Alcohol content (%)
Alcoholic extract of mouth wash
4.5
18
Alcoholic extract of mouthwash after boiling
5.1
Aqueous extract of mouthwash
5.8
Out of five samples of alcoholic mouth wash, turbidity

was observed after 8 months in two samples and 11 months
in other three samples. Whereas in aqueous mouth wash,
cloudiness and colour change were observed after 3 days.
This indicated that the durability period of alcoholic mouth
wash was higher (8-11 months) than that of aqueous mouth
wash (2-3days) at room temperature. But aqueous mouth
wash could be kept safe at 4C for 6-8 months.
In commercially available mouth wash, alcohol
content goes up to 27% and the pH ranges from 5-7 [13].
These two parameters were in acceptable ranges in newly
prepared mouth wash (Table 2). Results also showed that
aqueous extract of mouth wash containing natural
ingredients, exhibited better antibacterial activity when
compared to alcoholic extract. It had maximum sensitivity
towards Bacillus sp, while it had low sensitivity towards
the Klebsiella sp. Among the tested bacterial growth,
Staphylococcus aureus was only inhibited by both
preparations of mouth wash. All tested bacterial growth
was inhibited by the aqueous extract of mouth wash and
the positive control Chlorhexidine digluconate. But
alcohol alone (control) didnt inhibit the growth of any
tested bacteria (Table 1). This is due to less alcoholic
concentrations and the tolerance of test bacteria. Aqueous
natural mouth wash showed greater antibacterial activity
than alcoholic extracts of mouth wash. Hot extract method
was highly efficient for the extraction of antibacterial
compounds rather than tincture method. Long term use of
alcoholic mouth wash is not preferable, because of the
hazardous effects especially for children and causes
dehydration in mouth [14]. Recently the possibility that
the alcohol used in mouth wash acts as a carcinogen has
been raised [15]. Even though the durability period of
aqueous mouth wash was low at room temperature, it
showed greater range of antibacterial activity against test
bacteria and absence of alcohol. So this could be recommended rather than the alcoholic extract of mouth wash.
Further studies should be done clinically and test
the effectiveness of this Accmus aqueous extract of
mouth wash against oral diseases.
Conclusion
In both preparations of mouth wash pH and alcohol
content were in acceptable level. Staphylococcus aureus
growth was only inhibited by both mouth wash
preparations. Hot extraction method was efficient than
that of alcoholic extraction. Aqueous mouth wash showed
greater antibacterial activity against test bacteria and it
could be recommended with antibacterial activity rather

than the alcoholic extract of mouth wash.
References
1.
Goldberg S, Konis Y, Rosenberg. Effect of Cetylpyridinium

chloride on microbialadhesion to hexadecane and
polystyrene. Appl Environ Microbiol 1990: 1678-82.
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Giertsen E, Emberland H, Scheie AA. Effects of mouth

rinses with xylitol and fluoride on dental plaque and saliva.
Caries Res (1999); 33(1): 23-31.
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Lachenmeier DW, Keck WA, Sauermann A, Mildau G. Safety

assessment of alcohol containing mouth washes and oral
rinses. SOFWJ 2008; 134: 70-8.
4.
Farah C, Mclntosh L, McCullough M. Mouth washes.

Australian Prescriber 2009; 32: 162-4.
5.
National Institute of Science Communications. The Wealth

of India, Council of Scientific and Industrial Research, Vol
II, New Delhi 2001; 97.
6.
Muthaliyar M. Kunapadam (Moolikaivakuppu). 1936; I:

621-3.
7.
Pandey BP. Taxonomy of Angiosperms, S. Chand and

Company Ltd, 6th edition, Ram Nagar, New Delhi, 1997;
ISBN: 81-219-0932-5.
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Kugathasan KS. Check List of Some Plants of Botanical

and General Interest With Brief Descriptions, Field
Workcentre, Thondaimannaru, Jaffna. 2004; Bulletin no :16.
9.
Melntyre A. Herbs For Common Ailment, Gaia Books

Limited, UK. 1992; 13.
10. Chikitsai S. Sarabenthiravaiththiyamuraikal, SaraswathiMahal, Thanjavour. 1951; 89.

11. Nester W, Evans Roberts C, Pearsall N, Anderson G, Nester
T. Microbiology. A human perspective, WCB/Mc GrawHill, 2nd edition, 1998; ISBN-0-697-28602-9: 86-96.
12. Lino A, Deogracious O. The in vitro antibacterial activity
of Annona senegalensis, Securidacca longipendiculata and
Steanotaenia araliacea Ugandan medicinal plants. Afr
Health Sci 2006; 6: 31-5.
13. Lachenmeier DW, Keck-Wilhelm A, Sauermann A, Mildau
G. Safety assessment of alcohol-containing mouthwashes
and oral rinses. SOFW J 2008; 134: 70-8.
14. Cole P, Rodu B, Mathisen A. Alcohol-containing mouthwash
and oropharyngeal cancer: a review of the epidemiology. J
Am Dent Assoc 2003; 134(8): 1079-87.
15. Weaver C. Mouthwash linked to cancer. The Daily
Telegraph (News Ltd). 2009; http://www.news.com.au/
dailytelegraph/story/0,22049,24896583-5001021,00.html.
(retrieved 12 January 2009).
86
Review Paper
Anti rheumatic herbal compound drug Yi Shen Juan Bi (YJB) as selective

cytokines target in rheumatoid arthritis
Pathirage Kamal Perera1, Yunman Li2
Abstract
Rheumatoid arthritis (RA) is a chronic inflammatory
disease often resulting in increased morbidity, mortality
and disability. Cytokines regulate a broad range of
inflammatory processes that are implicated in the
pathogenesis of rheumatoid arthritis. In rheumatoid
joints, it is well known that an imbalance between pro
and anti-inflammatory cytokine activities favours the
induction of autoimmunity, chronic inflammation and
thereby joint damage. During recent decades a better
understanding of the pathogenesis of RA has led to the
development of new strategies for disease control which
have transformed the management of RA. However, none
of them are effective in curing rheumatoid arthritis.
Furthermore, the potentially greater efficacy of treatment
with TNF antagonists comes at a cost that is too high for
the majority of the world's population and with more side
effects. In this review we discussed about effective
compound herbal drug Yi Shen Juan Bi (YJB) derived
from traditional medicine as cytokine target in
rheumatology. According to our published research
findings YJB significantly ameliorate symptoms and
prevented severe arthritis development in rats. Our
studies showed that YJB significantly reduced the
production of IL-1, IL-6 and TNF- in vivo and in vitro.
These data indicate that YJB may have the potential to
regulate the immunomodulatory cytokines. So these
herbal compound drugs are more effective in the
treatment of RA. It is our hope that this kind of traditional
drugs can be developed to become new pharmaceutical
agents that can be used in cost and clinically effectively
for people suffering from rheumatic diseases.
Introduction
Rheumatoid arthritis (RA) is a common, chronic
disease, for which multiple pharmacotherapies are
generally applied. RA is a prevalent condition often leading

to a high burden of suffering in patients [1]. Conservative
treatment is mostly symptomatic and often associated with
adverse effects. Therefore, it is understandable that many
RA patients seek complementary and alternative medicine
(CAM) to manage their illness [1]. In the USA, about 60 to
90% of arthritis patients use CAM [2]. An Indian study
reported that around 40%of RA patients use either
Ayurvedic or homeopathic medicines or TCM alongside
conventional medicines [3]. According to the World Health
Organization (WHO), traditional herbal preparations
account for 30-50% of the total medicinal consumption in
China [4].
The suppression of auto immunity in RA can be
observed either as the induction of cell cycle arrest, which
slows down inappropriate or uncontrolled cell division,
or as the induction of apoptosis in stressed cells. Some
anti-inflammatory plant natural products have been found
to be very effective regulators of the cell cycle of
autoimmunity by targeting specific cell signaling
molecules, leading to apoptosis or cellular senescence.
Many anti-inflammatory plant natural products have
molecular signaling targets that can be potentially
employed for treatment of rheumatoid arthritis. A main
feature of a number of anti-inflammatory plant natural
products is their action on the suppression of
autoimmunity by upregulating key signaling molecules
like Bax, Bak, and Bid and the subsequent down
regulation of expression of various other key signaling
molecules such as NF-B, Bcl-2, and activate the
caspases, in the nucleus and/or cytoplasm, which
eventually induces apoptosis of target cells. Also most
plant natural compounds respond to inflammatory
mediators including IL-1, 4, 6, 8, 10, 12, 13, 17, 18, 21,
TNF-, TGF-, IFN-, VIP, iNOS, and cyclooxygenase2, prostaglandin E2.In this review we discussed herbal
medicine cytokines targets in rheumatology special
regards to Yi Shen Juan Bi (YJB) [5-8] (Table 1) .
Department of Dravyaguna Vignana, Institute of Indigenous Medicine, University of Colombo, Rajagiriya, Sri
Lanka.
2
Department of Physiology and Pharmacology, China Pharmaceutical University, Mailbox 207, Tongjiaxiang
24, Nanjing, Jiangsu, 210009, P. R. China.
Correspondence: Pathirage Kamal Perera, Department of Dravyaguna Vignana, Institute of Indigenous Medicine,
University of Colombo, Rajagiriya, Sri Lanka. E-mail: drkamalperera@yahoo.com. Received 20 August and revised
version accepted 15 November 2011.
Kamal Perera and Yunman Li. Anti rheumatic herbal compoundSLJIM 2011; 01(02): 86-90
Review Paper
Table 1: Ingredients of Yi Shen Juan Bi (patent
number: ZL200510040550) [5-8].
Ingredient
Composition
(g)
Rehmannia glutinosa
262.5
Eupolyphaga sinensis
210
Angelica sinensis
210
Bombyx batryticatus
210
Herba epimedii
210
Herba erodii
262.5
Buthus martensi
Corydalis yanhusuo
Scolopendra subspinipes
31.3
210
31.5
Panax ginseng
210
Polistes mandarinus
(stir-baking)
210
Cynanchum paniculatum
262.5
Rhizoma drynariae
Polygonum cuspidatum
31.5
262.5
Pyrola rotundifolia
31.5
Millettia reticulata
262.5
Zaocys Dhumnades
(stir-fried with wine)
210
Humulus scandens
262.5
Rehmannia glutinosa (dried)
210
The cytokine network in rheumatoid arthritis

Cytokines regulate a broad range of inflammatory
processes that are implicated in the pathogenesis of
rheumatoid arthritis. In rheumatoid joints, it is well known
that an imbalance between pro-and anti-inflammatory
cytokine activities favours the induction of autoimmunity,
chronic inflammation and thereby joint damage. However,
it remains less clear how cytokines are organized within a
hierarchical regulatory network, and therefore which
cytokines may be the best targets for clinical intervention
a priority. Analysis of cytokine mRNA and protein in
rheumatoid arthritis tissue revealed that many
proinflammatory cytokines such as TNF alpha, IL-1, IL-6,
GM-CSF, and chemokines such as IL-8 are abundant in all
patients regardless of therapy [9]. This is compensated to
some degree by the increased production of antiinflammatory cytokines such as IL-10 and TGF beta and
cytokine inhibitors such as IL-1ra and soluble TNF-R. In
rheumatoid joint cell cultures that spontaneously produce
IL-1, TNF alpha was the major dominant regulator of IL-1.
87
Subsequently, other proinflammatory cytokines were also

inhibited if TNF alpha was neutralized, leading to the new
concept that the proinflammatory cytokines were linked
in a network with TNF alpha at its apex. This led to the
hypothesis that TNF alpha was of major importance in
rheumatoid arthritis and was a therapeutic target. This
hypothesis has been successfully tested in animal models
of, for example, collagen-induced arthritis, and these
studies have provided the rationale for clinical trials of
anti-TNF alpha therapy in patients with long-standing
rheumatoid arthritis. Several clinical trials using a chimeric
anti-TNF alpha antibody have shown marked clinical
benefit, verifying the hypothesis that TNF alpha is of major
importance in rheumatoid arthritis. Retreatment studies
have also shown benefit in repeated relapses, indicating
that the disease remains TNF alpha dependent [9].
Immunomodulatory and anti-arthritic potential of
traditional medicine
Ayurveda, traditional Chinese medicine (TCM), and
other traditional systems are today yielding their theoretical
and experiential frameworks to investigation by modern
scientific techniques, applied mainly for the purpose of
illustrating the effectiveness of remedies that have been
developed over the centuries. In this context, the
underlying theoretical framework fades away, and the
tested substances become the focus of a new international
effort at preventive health care and disease treatment.
Herbal formulas developed today rely on a combination
of traditional and modern indications for the use of the
medicinal materials [10].
Arthritis has been a recognized medical condition
since ancient times, and the Chinese had developed
numerous formulas for its treatment. Chinese herbal
formulas were not specifically designed for either of the
two major types of arthritis defined today. The basis for
Chinese doctors differentiating arthritis into subgroups
was not the microscopic details of the pathology. Instead,
arthritis was divided into traditional medicine categories:
hot and cold types, upper and lower body involvement,
deficiency or excess syndrome, pain characteristics (such
as variability and severity), and whether the site of the
arthritis was fixed or "moving." Both rheumatoid arthritis
and osteoarthritis fall under the heading of bi syndrome, a
disorder of qi and blood circulation that leads to symptoms
of pain, numbness, swelling, and stiffness [11].
Rheumatoid arthritis fits most closely those syndromes
characterized by the Chinese as wind-damp invasion
affecting the joints. Osteoarthritis more closely fits the
syndrome of liver/kidney deficiency syndrome causing
weakness and stiffness in the legs with painful joints. In
China, syndromes similar to rheumatoid arthritis were an
area of special concern, generating considerable literature
on the subject, since the condition could arise suddenly
and could rapidly become severely debilitating [11].
Osteoarthritis, on the other hand, tended to be lumped
together with other disorders of aging, in which stiffness
88
Review Paper
and pain, especially of the legs, was considered just one

part of the gradual deterioration of body functions that
occurs with old age. As such, it is usually not the subject
of much discussion separate from antiaging therapies.
The closest traditional Chinese medicine term to
rheumatoid arthritis is fengshi bing which literally means
wind-damp disease [12-13]. The wind and damp factors
can complex with either cold or heat factors to yield
arthralgia. Almost all of the traditional approaches apply
to the complex involving cold factors rather than heat.
Gout, which has some characteristics in common with
arthritis, usually fits the cold-dominated category or the
cold-damp category of bi syndromes [12].
Chinese researchers have attempted to elucidate
how the herbs used in traditional arthritis formulas
alleviate the symptoms-from the modern viewpoint-by
carrying out numerous studies of the blood constituents
of patients [13]. According to studies that have been
carried out recently the mechanism of action that may be
dominant in the situations with good therapeutic results
is a reduction in the levels of pro-inflammatory cytokines,
such as interleukin-1 (IL-1), TNF etc [13]. The effect is
then to alter the levels of T-cells and the production of
activated antibodies and other components. In addition,
or as a result, the properties of the blood and its
circulation also change, with lowered sedimentation rate
and improved circulation to the extremities. The herbs
may also act on the prostaglandin synthesis and
degradation pathways, yielding a lower level of proinflammatory prostaglandins [13].
YJB as selective cytokine targeted anti rheumatic drug
The past studies evaluated anti-arthritic potential
of YJB in vivo and invitro rat models, which is very close
to its human counterpart. In these studies, we used
adjuvant arthritis (AA) induced and collagen induced
experimental rat models for our experiments. One of the
most imperative features of these models is chronic
synovitis, including inflammatory cell infiltration, panes
formation, and destruction of cartilage and bone erosion.
According to our research findings YJB significantly
ameliorate symptoms and prevented severe arthritis
development in rats [5, 6, 7, 8, 14].
To elucidate the effect of YJB on immunomodulatory
cytokines such as TNF-, IL-1 and IL-6, an ELISA assay
was performed. TNF- and IL-1 are considered key
mediators in the joint inflammation and in the destruction
of cartilage and bone in patients with RA [14].TNF- is a
pivotal mediator in inflammatory arthritis including RA
[15]. TNF- is an autocrine stimulator as well as a potent
paracrine inducer of other inflammatory cytokines such
as IL-1 and IL-6. The blockade and inhibition of TNF-
reduces the production of other inflammatory cytokines
in RA patients [16]. IL-6 is a proinflammatory cytokine
with a wide range of biological activities in immune
regulation, inflammation and oncogenesis [17]. IL-6 is
known to be responsible for the increase of serum gglobulin and the emergence of rheumatoid factors [18].
High levels of IL-6 have been observed in both sera and
synovial fluids from the affected joints of patients with
RA [19]. Our studies showed that YJB significantly
reduced the production of IL-1, IL-6 and TNF- [58].These data indicate that YJB may have the potential
to regulate the immunomodulatory cytokines. Further
our studies clearly confirmed that anti-arthritic property
of YJB substantiated lower TNF-, IL-1 production
capacity of macrophages in in vitro [5].
The contribution made by proinflammatory
cytokines in RA, such as tumour necrosis factor TNF-
and IL-1 has been validated in preclinical animal models
and in humans [20]. It is also well documented in human
RA and in animal models that IL-1 and TNF-
synergistically mediate synovitis and destruction of
cartilage and bone [21]. TNF- is an important regular
factor in inflammation and immunity response, which can
stimulate the synoviocyte and cartilage cells to
synthesize the PGE2 and collagenase causing synovium
and cartilage destruction as well as those of IL-1, IL-6,
and IL-8 [22]. Therefore it is one of the most important
factors in the cytokine network. In RA patients, IL-1 is
overexpressed in inflamed synovial tissue, in particular
in the lining layer and in sublining cells [23] and it is
elevated in draining lymph from affected joints [20].
Furthermore cartilage from arthritis patients' exhibits
upregulation of IL-1 mRNA as compared with normal
cartilage [24, 25]. In addition, increased production of
IL-1 in fibroblast like synoviocytes of susceptible
individuals may lead to a higher risk of developing severe
joint damage even in the absence of systemic
inflammation. In general, TNF- causes early joint swells
in RA, while IL-1 combining with the immune complex
leads to the cartilage erosion [25].
Considering these investigations it can be
concluded that TNF- and IL-1 have a pivotal role in
the pathogenesis of RA. Furthermore based on these
views, it can be pointed out that blocking of both TNF and IL-1 is necessary in the treatment of RA.
The study revealed that TNF- mRNA and IL-1
mRNA expression in synovial cells of model group was
significantly higher than that of normal rats. These are
correlated with above research findings in RA.
Furthermore pro-inflammatory cytokines, such as tumour
necrosis factor and interleukin-1, are expressed in the
arthritic joints in both AA rats and human rheumatoid
arthritis, and blockade of these molecules results in
amelioration of disease [26]. Our results confirmed that
YJB could significantly decrease the TNF- mRNA and
IL-1 mRNA expression in synovial cells. This may be
the one of the underlying mechanism that how YJB
ameliorate inflammation in RA. Therefore it is a promising
drug for the treatment of cytokine expression in vivo.
Review Paper
Conclusion
Taken together, our past results suggested that YJB
can be effectively applied to inflammatory and immune
diseases at the level of proinflammatory cytokines and
mediator regulation (see proposed mechanism of YJB's
activation in Figure 1).
Figure 1: The mechanisms of YJB activation proposed

here in scheme summarizes that the active components
, IL-
, IL-6, NO, PGE2, NFof YJB down regulate TNF-
B and COX-2 expression, which results in enhance anti
inflammatory and immunomodulatory action. YJB
potently induces the apoptosis of synovium, via ultimate
executioner caspase 3. YJB also down-regulates
cytochrome-c related Bcl-2 expressing and up-regulates
Bax expressing leading to triggered apoptosis cascade,
which results in enhance apoptosis in the RA synovium
and potentially limiting disease progression.'+': positive
effects '-': negative effects 'solid-line arrow': known
actions 'dotted-line arrow': our researches discovered
actions [5-8].
89
5.
Perera PK, Peng C, Xue L, Li Y, Han C. Ex vivo and in

vivo effect of Chinese herbal pill Yi Shen Juan Bi (YJB)
on experimental arthritis. J Ethnopharmacol 2011; 134:
171-5.
6.
Perera PK, Peng C, Li Y, Fang W, Han C. Immunomodulatory activity of a Chinese herbal drug Yi Shen Juan
Bi (YJB) in adjuvant arthritis. Indian J Pharmacol 2010;
42: 65-9.
7.
Perera PK, Peng C, Xue L, Li Y, Fang W, Han C. Effects of

Yi Shen Juan Bi (YJB) pill on experimental rheumatoid
arthritis. Chinese Journal of Natural Medicines 2010; 8:
57-61.
8.
Peng C, Perera PK, Li Y, Teng Q, Han C. Experimental

study on secondary affection of rat's collagen arthritis treated
with Yi Shen Juan Bi pill. Chinese Journal of Traditional
Medical Science and Technology 2010; 17(5): 389-90.
9.
Iain B, Schett M, Schett G. Cytokines in the pathogenesis

of rheumatoid arthritis. Nature Reviews Immunology 2007;
7: 429-42.
10. Hsu HY, Hsu CS. Commonly Used Chinese Herb Formulas
with Illustrations, rev. ed., Oriental Healing Arts Institute,
Long Beach, CA. 1980.
11. Hsu HY. Neuralgia, rheumatism, and gout and their Chinese
herb treatment. Bulletin of the Oriental Healing Arts Institute
1979; 4(4): 16-27.
12. Vangermeersch C, Sun Pei-lin. Bi-Syndromes, 1994 SATAS,
Belgium.
13. Guillaume G, Chieu M. Rheumatology in Chinese Medicine,
Eastland Press, Seattle, WA. 1996.
14. Ding CH, Li Q, Xiong ZY, Zhou AW, Jones G, Xu SY. Oral
administration of type II collagen suppresses proinflammatory mediators production by synoviocytes in
rats with adjuvant arthritis. Clin Exp Immunol 2003; 132:
416-23.
15. Firestein GS. Evolving concepts of rheumatoid arthritis.
Nature 2003; 423: 356-61.
16. Kitamura K, Nakamoto Y, Kaneko S, et al. Pivotal roles of
interleukin-6 in transmural inflammation in murine T cell
transfer colitis. J Leukoc Biol 2004; 76: 1111-7.
17. Bainbridge J, Madden L, Essex D, Binks M, Malhotra R,
Paleolog EM. Methionine aminopeptidase-2 blockade
reduces chronic collagen-induced arthritis: potential role
for angiogenesis inhibition. Arthritis Res Ther 2007; 9:
R127-R127.
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Amarasinghe APG, Park J, Dwivedi M, Ernst E. Ayurvedic

intervention for rheumatoid arthritis: a systematic review.
Focus on Alternative and Complementary Therapies 2003;
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Soeken KL, Miller SA, Ernst E. Herbal medicines for the

treatment of rheumatoid arthritis: a systematic review.
Rheumatology (Oxf) 2003; 42: 652-9.
3.
Sangha O. Epidemiology of rheumatic diseases.

Rheumatology (Oxf) 2000; 39 (Suppl 2): 3-12.
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World Health Organization: Traditional medicine (accessed

date: 25th August 2011). [http://www.who.int/mediacentre/
factsheets/fs134/en/].
18. Naka T, Nishimoto N, Kishimoto T. The paradigm of IL-6:

from basic science to medicine. Arthritis Res 2002; 4 (Suppl
3): S233-42.
19. Kim MJ, Kim HN, Kang KS, Baek NI, Kim DK, Kim YS,
Jeon BH, Kim SH. Methanol extract of Dioscoreae Rhizoma
inhibits pro-inflammatory cytokines and mediators in the
synoviocytes of rheumatoid arthritis. Int Immunopharmacol
2004; 4: 1489-97.
20. Dinarello CA. The IL-1 family and inflammatory diseases.
Clin Exp Rheumatol 2002; 20: S1-S13.
21. Choy EH, Isenberg DA, Garrood T, Farrow S, Ioannou Y,
Bird H, Cheung N, Williams B, Hazleman B, Price R,
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Yoshizaki K, Nishimoto N, Kishimoto T, Panayi GS.
Therapeutic benefit of blocking interleukin-6 activity with
an anti-interleukin-6 receptor monoclonal antibody in
rheumatoid arthritis: a randomized, double-blind, placebocontrolled, dose-escalation trial. Arthritis Rheum 2002; 46:
3143-50.
22. Choy EH, Panayi GS. Cytokine pathways and joint

inflammation in rheumatoid arthritis. N Engl J Med 2001;
344: 907-16.
23. Lipsky PE, van der Heijde DM, St Clair EW, Furst DE,
Breedveld FC, Kalden JR, Smolen JS, Weisman M, Emery
P, Feldmann M, Harriman GR, Maini RN. Infliximab and
methotrexate in the treatment of rheumatoid arthritis. AntiTumour Necrosis Factor Trial in Rheumatoid Arthritis with
Concomitant Therapy Study Group. N Engl J Med 2000;
343: 1594-602.
24. Olszewski WL, Pazdur J, Kubasiewicz E, Zaleska M,
Cooke CJ, Miller NE. Lymph draining from foot joints in

rheumatoid arthritis provides insight into local cytokine
and chemokine production and transport to lymph nodes.
Arthritis Rheum 2001; 44: 541-9.
25. Nadiv O, Beer Y, Goldberg M, Agar G, Loos M, Katz Y.
Decreased induction of IL-1 in fibroblast-like
synoviocytes: a possible regulatory mechanism
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44(12): 3147-54.
26. Alten R, Gram H, Joosten LA, van den Berg WB, Sieper
J, Wassenberg S, Burmester G, van Riel P, Diaz-Lorente
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Wright AM, Jung T. The human anti-IL-1 monoclonal
antibody ACZ885 is effective in joint inflammation
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patients with rheumatoid arthritis. Arthritis Res Ther
2008; 10(3): R67-R67.
Review Paper
91
Evidence based Ayurveda for revitalization of mental health

Nisha Ojha1, Abhimanyu Kumar1
Abstract
Mental health is an integral part of health. Mentally
healthy people are able to function well in home and
society and meet the ordinary demands of daily life. They
enjoy a positive quality of life and are free of disabling
symptoms of psychopathology. On the other hand, any
type of mental disorder is associated with a totally
disturbed life. According to the World Health Organization,
10% of the worlds population has some form of mental
disability and 1% suffers from severe incapacitating
mental disorders. In India community-based surveys
conducted during the past two decades showed that
the total prevalence of psychiatric disorder was around
5.8%. For children, only a few studies have reported a
prevalence rate ranging from 8.17-35.6% in India.
Although the current used drugs are the first choice
medication, however, these agents produce various
unacceptable side effects like, loss of appetite, stomach
aches/ cramps, headache, dizziness, irritability, drowsiness, staring, tics etc., which should be a matter of
concern in both adults and children. In this area, Ayurvedic
herbs having Medhya property may prove safe and
effective. Review of various experimental and clinical
studies offer clue to use different Medhya drugs,
judiciously for the management of psychiatric and
behavioural disorders.
Introduction
Mental health may be defined as a state of emotional
and psychological well being in which an individual is
able to use his or her cognitive and emotional capabilities,
function in society and meet the ordinary demands of
everyday life [1]. According to World Health Organization
mental health is defined as a being of well- being in which
the individual realizes his or her own abilities, can cope
with the normal stresses of life, can work productively
and fruitfully, and is able to make a contribution to his or
her community [2]. The World Health Organization states
that 10% of the world's population has some form of mental
disability and 1% suffers from severe incapacitating mental
disorders [3]. In India community-based surveys
conducted during the past two decades showed that the
total prevalence of psychiatric disorder was around 5.8%
[4]. Another study reports the prevalence rate for mental
disorders in India as 65.4 per 1000 population [5].
Mental health in children is defined by the
achievement of expected developmental cognitive, social
and emotional milestones and at the same time satisfying

social relationships and effective coping skills. Mentally
healthy children enjoy a positive quality of life; function
well at home, in school, and in their communities; and are
free of disabling symptoms of psychopathology [6]. For
children, only a few studies have reported a prevalence
rate ranging from 8.17-35.6% in India [7-11].
The most commonly occurring mental disorders are
anxiety disorders, mood disorders, personality disorders,
dementia and that of children as mentioned in DSM-IV,
include anxiety disorders, attention deficit and disruptive
behaviour disorders, autism and other pervasive disorders,
eating disorders, learning and communication disorders,
mood disorders, tic disorders etc. These disorders are
characterized by abnormal behaviour, thoughts, emotions
and relationship with others.
The current medications used in the treatment of
mental disorders including children incorporate
antipsychotic, antidepressants, anti anxiety drugs,
stimulants and mood stabilizing groups. Although these
drugs are the first choice medication, however, these
agents produce various unacceptable side effects like,
loss of appetite, stomach aches/cramps, headache,
dizziness, irritability, drowsiness, staring, tics etc., which
should be a matter of concern in both adults and children.
In this area, Ayurvedic herbs having Medhya property
may prove safe and effective. The drugs promoting
Medha (intellect) are termed as Medhya drugs.
Review of various experimental and clinical studies
offer clue to use different Medhya drugs, judiciously for
the management of psychiatric and behavioural disorders.
The review is taken from articles cited on Pubmed and in
MAPA (Medicinal and Aromatic Plant Abstracts) by using
the key words learning and memory, cognition, Bacopa,
Centella etc.
Nootropic activity
In a study, daily administration of Ashwagandha root
extract (50,100 and 200 mg/kg orally) for 6 days
significantly improved memory consolidation in mice
receiving chronic electroconvulsive shock (ECS)
treatment. Ashwagandha administered on day 7, also
attenuated the disruption of memory consolidation,
produced by chronic treatment with ECS. On the elevated
plus maze Ashwagandha reversed the scopolamine (0.3
Department of Bala Roga, National Institute of Ayurveda, Jaipur, 302002 India.
Correspondence: Prof. Abhimanyu Kumar, National Institute of Ayurveda, Jaipur, India. E-mail: ak_ayu@yahoo.co.in
Received 18 August and revised version accepted 15 November 2011.
Nisha Ojha and Abhimanyu Kumar. Evidence based Ayurveda SLJIM; 01:(02) 91-97
92
Review Paper
mg/kg) induced delay in transfer latency on day 1. On the

basis of these findings it is suggested that Ashwagandha
exhibits a nootropic like effect in naive and amnesic mice
[12]. A study indicate that treatment during postnatal
developmental stage with Centella asiatica extract can
influence the neuronal morphology and promote the higher
brain functions of juvenile and young adult mice [13].
Regeneration of nerves/ Induction of Axon-or Dendritic
outgrowth
Sub fractions of Centella asiatica ethanolic extract
were tested (100 microg mL 1) for neurite elongation in
the presence of nerve growth factor (NGF). Greatest
activity was found with a non-polar fraction (GKF 4).
Relatively polar fractions (GKF 10 and GKF 13) also
showed activity, albeit less than GKF 4. The findings
indicate that components in Centella ethanolic extract
may be useful for accelerating repair of damaged neurons
[14].
In a study, it was found that six of the 18 compounds
isolated from the methanol extract enhanced neurite
outgrowth in human neuroblastoma SH-SY5Y cells. In
Withanolide A treated cells, the length of NF-H-positive
processes was significantly increased compared with
vehicle treated cells, whereas, the length of MAP2positive processes was increased by Withanosides IV
and VI. These results suggest that axons are predominantly extended by Withanolide A, and dendrites by
Withanosides IV and VI [15].
Treatment with Withnolide A (WLA) isolated from
Ashwagandha, induced significant regeneration of both
axons and dendrites, in addition to the reconstruction of
pre and post synapses in the neurons. WLA (10 micro
mol Kg (-1) day (-1), for 13 days, p.o.) recovered A beta
(25-35) induced memory deficit in mice. At that time, the
decline of axons, dendrites and synapses in the cerebral
cortex and hippocampus was almost recovered. WLA is
therefore an important candidate for the therapeutic
treatment of neurodegenerative diseases, as it is able to
reconstruct neuronal networks [16].
Effect on cognitive function
In a study, Bacopa monnieri significantly improved
speed of visual information processing measured by the
IT task learning rate and memory consolidation compared
to placebo, with maximal effects evident after 12 weeks
[17].
In an experimental study, an extract of B. monnieri
was given to albino rats to measure its effect on three
newly acquired behavioural responses: brightness
discriminating, condition avoidance and continuous
avoidance. The facilitating effect of the Bacopa was clearly
discernible in all three learning responses, augmenting
both the rats cognitive function and mental retention
capacity. The rats learned faster, retained more of what
they had learned, and remembered it longer. The chemical
constituents responsible for the facilitating effect of

Bacopa on learning schedules were identified as a mixture
of bacosides A and B. The bacosides also enhanced vital
protein activity and produced an increase in protein
synthesis in the hippocampus, a part of the brain that is
important for long-term memory [18].
In an open 4 week trial of Bacopa in 35 patients with
anxiety neurosis 12g/day of dried Bacopa herb was given
in the form of syrup. Significant improvement in anxiety
(P<0.05), concentration (P<0.05) and immediate memory
span (P<0.01) were seen as a result of the treatment. Work
related mental fatigue, measured as total work output and
errors committed per unit time, also improved significantly
with Bacopa treatment (P<0.001). Improvements were also
seen in symptoms such as insomnia, headache, palpitation
and irritability [19]. To investigate the effect of Bacopa in
school children aged 6-8 years, 40 children were given
Bacopa syrup equivalent to 1 g dried herb daily for 3
months, in a single-blind design. Immediate memory,
perception and reaction/ performance times improved with
Bacopa treatment [20].
Neonatal rat pups (7 days old) were given different
doses of fresh leaf juice of C. asiatica (CeA) orally for
different period of time. These rats were then subjected to
spatial learning (T-Maze) and passive avoidance tests
along with the age matched normal saline control rats.
The result showed improvement in spatial learning
performance and enhanced memory retention in neonatal
rats treated with higher doses [21].
In a study on the effects of Brahmi (B. monnieri) on
human memory, seventy six adults aged between 40 and
65 years took part in a double blind randomized, placebo
control study in which various memory functions were
tested and levels of anxiety measured. The results showed
a significant effect of the B. monnieri on a test for the
retention of new information. Follow up tests showed that
the rate of learning was unaffected suggesting that B.
monneiri decreases the rate of forgetting of newly acquired
information [22]. Bacopa has also demonstrated a
significant memory promoting effect in animal models of
Alzheimers disease [23].
In an experimental study, fresh C. asiatica plant
extract was given orally to rat pups (n=5), from P7-P49 (6
weeks, 2 ml/kg/day). These and age matched normal
control (n=5) rats were subjected to learning tests in Tmaze and passive avoidance test. Following this, rats were
sacrificed and amygdaloid nucleus was processed for Golgi
staining. Results showed a significant increase in the
percent correct response (control: 86.44 + 2.33 percent Vs
Expt. 93.44 + 3.90 percent) in plant extract treated rats.
Passive avoidance retention test revealed a significantly
memory retention, dendritic intersection was significantly
increased at all concentric circles, except at 100 micron.
Dendritic branching points also significantly increased in
the inner three zones. These results indicate a correlation
between improved learning capacity and increased
Review Paper
dendritic arborization in amygdaloid nucleus. This may
be the neural basis for enhanced learning in Centella
asiatica treated rats [24].
A study undertaken to asses the potential of
Nordostachys jatamansi as a memory enhancer, elevated
plus maze and the passive avoidance paradigm were
employed to evaluate learning and memory parameters.
Three doses (50, 100, and 200 mg./kg. p.o.) of an ethanolic
extract of N. jatamansi were administered for 8 successive
days to both young and aged mice.
The 200 mg/kg dose of N. jatamansi ethanolic extract
significantly improved learning and memory in young mice
and also reversed the amnesia induced by diazepam (1
mg/kg, i.p.) and scopolamine (0.4mg/kg.i.p.). Furthermore,
it also reversed aging induced amnesia due to natural aging
of mice. Hence, N. jatamansi might prove to be a useful
memory restorative agent in the treatment of dementia
seen in elderly persons [25].
A study undertaken with the objective of studying
the effect of Tinospora cordifolia (Tc) on learning and
memory in normal rats and on cyclosporine induced
memory deficits, both alcoholic and aqueous extract of
T. cordifolia enhanced the cognition in normal rats as
were seen in behavioural tests Hebb William maze and
the passive avoidance task [26].
To investigate the effects of Glycyrrhiza glabra, on
learning and memory, the elevated plus- maze and passive
avoidance paradigm were employed to evaluate learning
and memory parameters. Three doses (75, 150 and 300 mg/
kg p/o) of aqueous extract of G. glabra were administered
for 7 successive days in separate groups of mice. The
dose of 150 mg/kg of the aqueous extract of liquorice
significantly improved learning and memory of mice.
Furthermore, this dose reversed the amnesia induced by
diazepam 1mg/kg i.p.), scopolamine (0.4 mg/kg i.p) and
ethanol (1mg/kg i.p) [27].
The effect of Withania somnifera extract prepared
by two different methods were assessed on behavioral
parameters using open field exploratory behavior, behavior
despair and passive avoidance tests and were compared
in young and old stressed Wistar rats. W. somnifera
extracts prepared with 50% methanol and solvent
containing water, ghee and honey were administered orally
as fine suspension, during the shock period. The results
revealed that stress produced depression anxiety and
retention deficit in young and old rats. Administration of
W. somnifera methanolic extract 250 mg/ kg during shock
period in young and old rats attenuated the stress-induced
depression and enhanced memory. W. somnifera traditional
extract 250mg/kg produced memory enhancement in both
control and stressed young and old rats [28].
Isolated constituents of W. somnifera (Sitoindosides
VII-X and Withaferine A) increased cortical muscarinic
acetylcholine receptor capacity partly explaining the
cognition enhancing and memory improving effects
traditionally attributed to Ashwagandha [29].
93
In a study, one half of a group of 40 healthy children

(ages 6-8) were given Bacopa in a syrup base three times
a day ( a total of 1.05 g/day) over the course of four weeks,
while the other half were given a placebo. Those children
taking Bacopa were superior in matters of speed and
accuracy in solving maze problems. Overall, these
improvements vitalized the childrens efficacy and their
propensity to choose exploratory behaviour and to opt
for novel experiences in preference to familiar ones [30].
Effect in ADHD
In a study 36 children in the 8-10 year age group were
selected for a double blind, randomized trial. 19 were given
50 mg of Bacopa twice daily, 17 others received placebo.
After 12 weeks of treatment, the children were subjected
to a battery of specialized tests. The data revealed a
significant improvement in the areas of sentence repetition,
logical memory and pair associative learning (matching
things that go together, e.g. test and grade) in all 19
ADHD children who took Bacopa [31].
A study to test the efficacy of Bacopa on children
for six weeks, 50 normal school children split into two
groups were given either Bacopa or placebo. At the
conclusion, they were evaluated for attention,
concentration, and memory. Bacopa was shown to improve
mean reaction time (auditory and visual) significantly [32].
Effect on behaviour
In an experimental study, rats were individually trained
in a simple T-maze until they reached a predetermined level
of performance. They were then divided into three groups
and given either nothing, diazepam (Valium), or Bacopa.
At the end of 10 days, they were evaluated by repeating
the T-maze trial. Those animals given Bacopa, showed
remarkable learning and memory enhancement compared
with the control and valium groups. Furthermore, the
neurochemical content of their brain tissue showed an
increase in the level of serotonin. Serotonin has been
identified with improved spatial memory as well as
anxiolytic benefits [33].
Antidepressant activity
In a study, B. monneiri extract given in the dose of 20
and 40 mg/kg, orally once daily for 5 days was found to
have significant antidepressant activity in forced swim
and learned helplessness models of depression and was
comparable to that of imipramine [34].
A 15 day treatment with N. jatamansi resulted in a
significant increase in the levels of NE, DA, 5-HT, 5-HIAA
and GABA. These data indicate that the alcoholic extract
of the roots of N. jatamansi causes an overall increase in
the levels of central monoamines and inhibitory amino
acids [35].
In an experimental study a bioassay-guided isolation
of the ethanol extract from the fruits of Piper longum
yielded, a known piperidine alkaloid, piperine, which
94
Review Paper
showed an inhibitory effect against monoamine oxidase

(MAO). The results suggest that piperine possesses
potent antidepressant-like properties that are mediated in
part through the inhibition of MAO activity and therefore
represent a promising pharmaco therapeutic candidate as
an antidepressant agent [36].
Anxiolytic/ antistress activity
The bioactive glycowithanolides (WSG), isolated
from W. somnifera roots WSG (20 and 50 mg/kg) was
administered in rats orally once daily for 5 days and the
results were compared by those elicited by the
benzodiazepine lorazepam (0.5mg/kg, i.p.), for the
antidepressant investigations. WSG induced an anxiolytic
effect, comparable to that produced by lorazepam, in the
elevated plus-maze test, social interaction and feeding
latency in an unfamiliar environment. WSG also exhibited
an antidepressant effect, comparable with that induced
by imipramine, in the forced swim induced behavioral
despair, and learned helplessness, tests [37].
In a study, mice received varying doses [10-300mg/
kg i.p.] of hydroalcoholic extract of roots and rhizomes of
G. glabra and anxiolytic activity was assessed using
different paradigms like elevated plus maze, foot shockinduced aggression and amphetamine induced stereotypy.
In all the animal models of anxiety, lower doses of
hydroalcoholic were more effective in alleviating anxiety.
The extract and standard anxiolytic agents increased
duration of occupancy of mice in open arm, increased
latency to foot shock-induced aggression and reduced
number of fighting bouts and delayed the onset of
amphetamine induced grooming, biting, sniffing and
repetitive head movements [38].
The antistress effect of bacosides of Brahmi (B.
monneiri) was studied in adult male Sprague Dawley rats
by administering oral doses of 20 and 40 mg/kg for 7
consecutive days. The data indicated that Bacopa has
potential to modulate the activities of HSP 70, P450 and
SOD (super oxide dismutase) thereby possibly allowing
the brain to be prepared to act under adverse conditions
such as stress [39].
A number of herbal drugs mostly in the form of their
extracts (holistic approach) or in some, as active principles
isolated from them, were evaluated for their antistress
activity by a number of tests. W. somnifera, Ocimum
sanctum, T. cordifolia, C. asiatica, G. glabra were reported
with encouraging results [40].
Neuroprotective effect
The effect of an aqueous extract of C. asiatica (100,
200 and 300 mg/kg for 21days) was evaluated in i.c.v.
STZ induced cognitive impairment and oxidative stress
in rats. The findings indicated that an aqueous extract of
C. asiatica is effective in preventing the cognitive
deficits, as well as the oxidative stress, caused by i.c.v.
STZ in rats [41].
The protective effect of N. jatamansi (NJ) on

neurobehavioural activities was studied in middle cerebral
artery (MCA) occlusion model of acute cerebral ischaemia
in rats. All the alternations induced by ischemia were
significantly attenuated by 15 days pretreatment of NJ
(250 mg/kg p.o.) and correlated well with histopathology
by decreasing the neuronal cell death following MCA
occlusion and reperfusion [42].
CNS Depressant/ sedative effect
The methanol extract of the whole plant of
Shankhapushpi, Convolvulus microphyllus sieb ex spreng
(convolvulaceae), was found to produce alternations in
the general behaviour pattern, reduction in spontaneous
motor activity, hypothermia, and potentiation of
phenobarbitone-sleeping time, reduction in exploratory
behavioural pattern and suppression of aggressive
behaviour. The extract also showed an inhibitory effect
on conditioned avoidance response and antagonism to
amphetamine toxicity. The findings explicitly suggested
that the whole plant extract of C. microphyllus possesses
a potential CNS depressant activity [43].
Effect of chronic administration of ethanolic extract
of Acorus calamus (AC) was studied on spontaneous
electrical activity and monoamine levels of brain. AC
seemed to exert its depressive action by changing electrical
activity and by differentially altering brain monoamine
levels in different brain regions [44].
A sedative action and potentiation of barbiturate
effect (increased sleeping time, reduced body temperature)
was observed in small animals (mice, rats, rabbits and cats)
following intraperitoneal adminstration of the aquous and
ethanolic extracts of both European and Asian varities of
A. calamus [45].
CNS stimulating action
The effects of a 50% ethanol extract of the root of
Plumbago zeylanica were investigated on locomotor
behavior and central dopaminergic activity in rats. The
results showed that the extract of the root of P. zeylanica
specifically enhanced the spontaneous ambulatory activity
without inducing stereotypic behaviour. The neurochemical estimations revealed elevated levels of DA and
its metabolite homovanillic acid (HVA) in striatum
compared with the control rats. These behavioural and
biochemical results indicated stimulatory properties of the
extract of the root of P. zeylanica, which may be mediated
by dopaminergic mechanisms in the rat brain [46].
Discussion
Review of the various clinical and experimental
studies of different Ayurvedic Medhya drugs reveals that
these drugs possess nootropic, cognition enhancing, antistress, anxiolytic, antidepressant, anticonvulsant, CNS
depressant, sedative and neuroprotective effect.
Review Paper
Ashwagandha and Centella possess nootropic
activity. Bacopa and Ashwagandha have the potential
for corrective effect in cognitive deficit, while Centella
can influence the neuronal morphology and can thereby
promote higher brain functions.
The drugs C. asiatica and W. Somnifera
(Ashwagandha) are useful for repair of damaged neurons
and in neurodegenerative disorders as these drugs have
the capacity to reconstruct the neuronal networks.
Regarding the effect on cognition, the drugs Bacopa,
Centella, N. jatamansi, T. cordifolia, G. glabra and W.
somnifera may be useful. Bacopa has potential for
revitalization of intellectual functions and may improve
the higher order cognitive functions such as learning and
memory. It has also showed effect on positive behavior
modification by increasing the level of serotonin. Centella
can induce memory retention an improved learning capacity
by increasing dendritic arborization in amygdaloid
nucleus.
N. jatamansi is useful as memory restorative agent
as it facilitates cholinergic transmission in the brain. Both
T. cordifolia and G. glabra are helpful in improving the
learning capacity and memory restoration. W. somnifera,
by increasing cortical muscarinic acetylcholine receptor
capacity, can enhance memory.
95
pattern, reduction in spontaneous motor activity,

hypothermia, and potentiation of phenobarbitonesleeping time, reduction in exploratory behavioural pattern
and suppression of aggressive behaviour. A. calamus
seems to exert its depressive action by changing electrical
activity and by differentially altering brain monoamine
levels in different brain regions. P. zeylanica possesses
CNS stimulant activity.
Conclusion
The studies indicate that Ayurvedic medhya drugs
act in different way on the brain and different drug have
specific action. Thus these drugs have potential to
improve the overall mental system. Studies also reveal
that Brahmi (B. monnieri) is the best intellect promoting
drug which can prove safe and effective in promoting
mental health in children. The review provides a clear view
about the specific properties of different medhya drugs
thus ther judicious use will be very much helpful for the
management of mental and behavioural disorders of adults
and children.
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and SOD (super oxide dismutase) thereby possibly
allowing the brain to be prepared to act under adverse
conditions.
C. asiatica by its potentiation of cellular oxidative
defense mechanism acts as neuroprotective drug.
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Guidelines
Guidelines for authors

accepts original research reports in English in all areas of
basic scientific and clinical research on indigenous/
traditional system of medicine, medicinal plants, Ayurvedic
pharmaceutical science etc. Further the editors encourage
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99
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Volume 01

ISSN 2012 9238

Sri Lanka Journal of Indigenous Medicine

Peer reviewed research publication of the

INSTITUTE OF INDIGENOUS MEDICINE

Sri Lanka Journal of Indigenous Medicine (SLJIM)

Prof. W. D. Rathnasooriya PhD

Prof. Jayantha Welihinda PhD

Dr. A. H. M. Mawjood PhD

Dr. D. P. A. Dissanayaka MPhil

EDITORIAL BOARD MEMBERS

Senior Lecturer, Department of Dravya Gunavignana

Prof. S. Bhavani MSAM

Dr. D. M. R. B. Dissanayaka MSAM

Dr. Praneeth Abesundara PhD

Prof. Ikhlas Khan PhD

Prof. Manjari Dwivedi PhD

Prof. M. S. Bhagel PhD

Senior Lecturer, Department of Basic Principles

Prof. Abhimanyu Kumar PhD

Prof. M. Shahu PhD

Prof. R. R. Dwivedi PhD

Sri Lanka Journal of Indigenous

The herb is mainly used to promote intellect and

Brahmine and Herpestine are major alkaloid

The original paper on page 55 and review paper

Single issue: Rs. 300/= (Local)

Sri Lanka Journal of Indigenous Medicine (SLJIM)

S M S Samarakoon, S K M K Herapathdeniya, H M Chandola, B Ravishankar

Study of the efficacy of an ayurvedic treatment regimen on balaka pakshaghatha

Saroja Weerakoon, A P G Amarasinghe

Clinical efficacy of Dashamoola Taila Matra Vasti on management of primary dysmenorrhoea

Kaumadi Karunagoda, Shilpa Donga, Lakshmi Priya Dei

E Christy Jeyaseelan, S Tharmila, A C Thavaranjit

N D N Priyadarshani, M K T K Amarasinghe, S Subasinghe, I R Palihakkara, H K M S Kumarasinghe

Anju P Ramachandran, M Shyam Prasad, Vijay Kumar, B K Ashok, B Ravishankar, H M Chandola

S Tharmila, T Thileepan, A C Thavaranjit, R Srikaran

Pathirage Kamal Perera, Yunman Li

Evidence based Ayurveda for revitalization of mental health

Nisha Ojha, Abhimanyu Kumar

Guidelines for authors

Experimental evaluation of gastroprotective and adaptogenic activity

Ayurvedic physicians. However, no report on the

Materials and Methods

Dose selection and schedule: The classical dose of AR in

Effect of drugs on stress-induced ulcer was evaluated

Samarakoon et al. Experimental evaluation of gastro protectiveSLJIM 2011; 01 (02): 51-54

0.27+0.7+1.4 16.470 0.530 **

*p<0.05 **p<0.001 Vs stress control (unpaired t test); p<0.05 Vs

Administration of vehicle and test drug significantly

Effect of AR and vehicle on ulcer index

way Anova - F value 102.50; p<0.001: p<0.05 for VC and

Vs vehicle control (Unpaired t test)

Administration of vehicle and test drug significantly

promotion of health by revitalizing the metabolism and

Samarakoon et al. Experimental evaluation of gastro protectiveSLJIM 2011; 01 (02): 51-54

be concluded that AR is having significant adaptogenic

Kirkwood T, Ebrahim S, Kalache A. Mechanisms of Ageing

Hamilton IS. The Psychology of Ageing: An Introduction,

Acharya JT. Sushruta Samhita, Chaukmbha Surabharati

Acharya JT. Charaka Samhita, Chaukambha Prakashan,

Acharya JT. Charaka Samhita, Chaukambha Prakashan,

Acharya JT. Charaka Samhita, Chaukambha Prakashan,