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Global Journal of Research on Medicinal plants & Indigenous medicine's - May 2013 issue
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An International, Peer Reviewed, Open access, Monthly E-Journal
ISSN 2277 – 4289 www.gjrmi.com
Editor-in-chief
Dr Hari Venkatesh K Rajaraman
Managing Editor
Dr. Shwetha Hari
Administrator & Associate Editor
Miss. Shyamala Rupavahini
Advisory Board
Prof. Rabinarayan Acharya Dr. Dinesh Katoch
Dr. S.N.Murthy Dr. Mathew Dan Mr. Tanay Bose
Dr. Nagaraja T. M. Prof. Sanjaya. K. S. Dr. Narappa Reddy
Editorial board
Dr. Kumaraswamy Dr. Madhu .K.P
Dr. Sushrutha .C.K Dr. Ashok B.K.
Dr. Janardhana.V.Hebbar Dr. Vidhya Priya Dharshini. K. R.
Mr. R. Giridharan
Honorary Members - Editorial Board
Dr Farhad Mirzaei Mr. Harshal Ashok Pawar
INDEX – GJRMI, Vol.2, Iss. 5, May 2013
MEDICINAL PLANTS RESEARCH
Bio-Chemistry NATURAL PLANT PRODUCT BERBERINE/CISPLATIN BASED RADIOTHERAPY FOR
CERVICAL CANCER: THE NEW AND EFFECTIVE METHOD TO TREAT CERVICAL CANCER
Komal, Singh Mayank, Deshwal Vishal kumar 278–291
Bio-technology PHYTOCHEMICAL SCREENING OF SECONDARY METABOLITES OF EUPHORBIA
NERIIFOLIA LINN.
Chouhan Leela, Bhatt Shashank, Dhyani Suresh 292–297
Ethno-Medicine
TRADITIONAL MEDICINES OF HERBAL ORIGIN PRACTICE BY THE ADI TRIBE OF EAST
SIANG DISTRICT OF ARUNACHAL PRADESH, INDIA
Das Moushumi, Jaishi Anju, Sarma Hirendra N 298–310
Bio-technology PRELIMINARY PHYTOCHEMICAL SCREENING OF SECONDARY METABOLITES OF
ADHATODA VASICA NEES. FLOWERS
Chouhan Surksha, Bhatt Shashank, Dhyani Suresh 311–316
Review Article
HERBAL MEDICINES FOR DEPRESSION AND ANXIETY: A COMPREHENSIVE STATE OF
THE ART REVIEW
Patel Shanti, De Sousa Avinash 317–336
Review Article A BRIEF REVIEW ON NONI (MORINDA CITRIFOLIA L.) - A HERBAL REMEDY FOR BETTER
HEALTH
Patel Swetal, Krishanamurthy R 337–347
INDIGENOUS MEDICINE
Ayurveda – Dravya Guna DEVELOPMENT OF RANDOM AMPLIFIED POLYMORPHIC DNA MARKERS FOR
AUTHENTICATION OF RIVEA HYPOCRATERIFORMIS (DESR.) CHOISY
Borkar Sneha D, Naik Raghavendra, Harisha C R, Acharya R N 348–356
Ayurveda – Dravya Guna
ANTIMICROBIAL EVALUATION OF CROTON ROXBURGHII BALAK. (EUPHORBIACEAE)
STEM BARK
Patel Esha, Padiya RH, Acharya RN 357–364
Ayurveda – Kaya Chikitsa
AN ESTIMATION OF HUMIC SUBSTANCES IN AN AYURVEDIC HERBOMINERAL DRUG
SHILAJATU (ASPHALTUM) AS PART OF PHYTO-PHARMACEUTICAL STANDARDIZATION
Akarshini A M, Renuka, Shukla V J, Baghel M S 365–373
Ayurveda – Shalya Tantra
MANAGEMENT OF ARBUDA (CANCER) WITH HERBOMINERAL FORMULATION - A PILOT
STUDY
Mahanta Vyasadeva, Dudhamal T S, Gupta S K 374–379
Ayurveda – Review Article - Moulika Siddhanta
IMPORTANCE OF UPAYOGASAMSTHA (DIETETIC RULES) IN RELATION TO DIGESTION OF
THE FOOD
Avhad Anil D, Vyas H A, Dwivedi R R 380–385
Ayurveda – Review Article - Moulika Siddhanta
CONCEPT OF VYADHIKSHAMATVA (IMMUNITY) AND ITS RELATIONSHIP WITH BALA
(VITAL STRENGTH)
Sharma Mahesh Kumar 386–391
Ayurveda – Review Article - Dravya Guna
A REVIEW ON VARIETIES OF ARKA - CALOTROPIS PROCERA (AITON) DRYAND. AND
CALOTROPIS GIGANTEA (L.) DRYAND.
Poonam, Gaurav Punia 392–400
COVER PAGE PHOTOGRAPHY: DR. HARI VENKATESH K R, PLANT ID – FLOWER & IMMATURE FRUIT OF NONI (MORINDA
CITRIFOLIA. L. ), OF THE FAMILY RUBIACEAE PLACE – KOPPA, CHIKKAMAGALUR DISTRICT, KARNATAKA, INDIA
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 278–291
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
NATURAL PLANT PRODUCT BERBERINE/CISPLATIN BASED
RADIOTHERAPY FOR CERVICAL CANCER: THE NEW AND EFFECTIVE
METHOD TO TREAT CERVICAL CANCER
Komal1, Singh Mayank
2, Deshwal Vishal kumar
3*
1, 2
Department of Biochemistry, All India Institute of Medical Science, New Delhi, India. 1, 3
School of Life Sciences, Singhania University, Pachri Bari, Rajasthan, India.
*Corresponding Author: E-mail: [email protected]; Mobile: +919897538555
Received: 28/03/2013; Revised: 12/04/2013; Accepted: 15/04/2013
ABSTRACT
Cervical cancer is the site of excessive inflammation which leads to extensive DNA damage and
thus promotes carcinogenesis. Existing treatment regime for cervical cancer is radiotherapy along
with platinum based drugs like cisplatin and carboplatin but it is associated with various side effects
to normal cells and problem of radio-resistance. Berberine is a natural chemo-preventive agent
extracted from Berberis aristata that has been shown to suppress and retard carcinogenesis by
inhibiting inflammation. In this study we compared the cisplatin based radiotherapy with a
Berberine/cisplatin based radiotherapy in cervical cancer in vitro. Treatment of cervical cancer cell
lines SiHa and CaSki with Berberine/cisplatin combination followed by treatment by ionizing
radiation (IR) resulted in increased apoptosis in comparison to cisplatin based radiotherapy. The
combination therapy of Cisplatin/Berberine/IR resulted in upregulation of pro apoptotic proteins like
Bax, p73 and down regulation of anti apoptotic Bcl Xl, inflammatory Cox 2, Cyclin D1 accompanied
by increase in activity of Caspase -9 and -3. Reduction in Telomerase activity was also seen in all the
HPV positive cells.
KEYWORDS: Cervical cancer, Radiotherapy, Berberine, Cisplatin
Research article
Cite this article:
Komal, Singh M, Deshwal V K (2013), NATURAL PLANT PRODUCT BERBERINE/CISPLATIN
BASED RADIOTHERAPY FOR CERVICAL CANCER: THE NEW AND EFFECTIVE METHOD TO
TREAT CERVICAL CANCER, Global J Res. Med. Plants & Indigen. Med., Volume 2(5): 278–291
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 278–291
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
INTRODUCTION
Cervical cancer remains one of the major
cancers amongst women worldwide with a high
rate of mortality (Ciesielska et al., 2012).
Radiotherapy in combination with cisplatin
remains the treatment of choice in majority of
cases in which the cancer is locally advanced.
Currently the chemotherapeutic drugs cisplatin
and 5-Fluorouracil are used as radio-sensitizers
along with ionizing radiation (IR) for the
treatment of cervical cancer (Rosa et al., 2012).
Nevertheless, therapeutic results are far from
optimal, so new and safer therapeutic
combinations need to be investigated which
specifically target cervical cancer cells with
minimal toxicity to normal tissue. Although
chemo radiotherapy is more effective as
compared to radiotherapy alone, it is associated
with dose limiting toxicities like
gastrointestinal and hematological toxicities
(Tan et al., 2012). Natural products offer an
excellent alternative for therapeutic use as
opposed to synthetic compounds because of
their relatively well established safety profile
(Deshwal, 2012; Kumar et al., 2012; Makhloufi
et al., 2012). Several natural products are being
tested as potential radio sensitizers. Berberine
is a natural compound that allows prevention,
suppression and retardation of carcinogenesis.
Berberine [1, 7-bis- (4 –Hydroxy-3-
methoxyphenyl) -1, 6 heptadiene -3, 5-dione] is
a major constituent of product extracted from
the rhizome of the plant Berberis aristata found
in South and Southeast tropical Asia. Berberine
(Plate 1) has been shown to be a potent chemo-
preventive agent inhibiting tumor progression
against skin, oral, intestinal, breast, colon and
prostate cancer (Diogo et al., 2011). Berberine
has been shown to confer radiosensitizing
effect in prostate cancer cells, squamous cell
carcinomas (Vuddanda et al., 2010; Tillhon et
al., 2012) and recently in HeLa and SiHa,
human cervical cancer cells (Javvadi et al.,
2008). The major problem with cervical cancer
is that the cancer cells become increasingly
radioresistant due to activation of various
antiapoptotic genes/cascades resulting in
therapy failure, and the standard chemo-
radiotherapy regime is unable to address this
problem (Aggarwal et al., 2006). The goal of
the present study was to compare the standard
chemo-radiotherapy regime comprising of
cisplatin/IR with the cisplatin/Berberine/IR
combination therapy and assess its effect on
protein which confers radio resistance to
cervical cancer cells in vitro.
Plate 1: Chemical structure of Berberine
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 278–291
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
MATERIAL AND METHODS
Cell culture and chemicals
Human cervical cancer SiHa and Ca Ski
cells were obtained from National Centre for
Cell Sciences, Pune, India and were maintained
in either Dulbecco’s modified Eagle’s medium
or RPMI1640 (Sigma, USA) supplemented
with 10% (v/v) heat-inactivated fetal bovine
serum (Hyclone), antibiotics, in a humidified
atmosphere of 95% air and 5% CO2 at 37ºC.
Cells were exposed to varying Ionizing
radiation (IR). Antibodies against p73, Bcl xl,
Bax, Cyclin D1, AIF and Cox 2 as well as
secondary AP conjugated antibodies were
obtained from Santa Cruz, USA. Berberine was
obtained from Sigma, USA
Flow cytometry
Cells (1 × 104
cells) were treated with
50 μM and 75 μM Berberine for 24 hrs and
then harvested. Cells were fixed in 70% ethanol
and left overnight at −20ºC. Cells were then
washed with PBS and incubated in staining
solution (20 μg/ml propidium iodide, 50 μg/ml
RNAse, 0.1% Triton X-100 and 0.1 mM
EDTA) for 2 hrs at 4ºC in dark. The DNA
content of the cells was measured by flow
cytometer (Becton Dickenson, USA) using
Diva software.
Assay of telomerase activity
This was measured using the PCR-ELISA
kit based on TRAP (Telomerase repeat
amplification protocol) assay (Roche Molecular
Bio-chemicals, Germany). The samples were
lysed and an aliquot containing 2μg protein was
used for the assay. Telomerase positive
embryonic kidney cell line (HEK-293) was
used as positive control while heat inactivated
HeLa extract was used as negative control. The
telomerase activity was detected and expressed
as relative units (RU) (Khanna et al., 2003).
Western blot analysis
The level of expression of various proteins
was determined in control and treated cells by
Western blotting as described previously
(Singh et al., 2007). Briefly, cells were washed
twice in PBS (Phosphate buffered saline) and
lysed in RIPA lysis buffer. Total protein was
determined by the Bradford assay. Equal
amount of protein was loaded and run on 10–
15% SDS-polyacrylamide gel. The proteins
were transferred to a nitrocellulose membrane.
The membrane was blocked with 5% BSA
(Bovine serum albumin), followed by
hybridization with respective primary and
secondary antibody. Final detection was
performed with BCIP/NBT BCIP (5-bromo-4-
chloro-3'-indolyphosphate p-toluidine salt/
NBT (nitro-blue tetrazolium chloride),
substrate (Promega, USA). The bands were
analyzed and quantified using ά image scanner
densitometer and normalized with β actin
control. The density of control was taken as 1
and results of treatment were expressed in
relation to the control as relative unit (RU).
Assay of Caspase -3, -9 activities
Caspases-3 and -9 activity were measured
by the direct assay for Caspase enzyme activity
in the cell lysate using synthetic fluorogenic
substrate (Ac-DEVD-AFC; substrate for
caspase 3; MBL Bioscience, USA; Ac-LEHD-
AFC, substrate for Caspase 9; Genotech, USA)
as described by the manufacturer. Amount of
fluorogenic AMC/AFC moiety released was
measured using a spectro-fluorimeter
(ex.380 nm, em.420–460 nm for Caspase -3;
ex.400 nm, em.490–520 nm for Caspase-9).
The results are expressed in arbitrary
fluorescence units/mg protein (Singh and
Singh, 2008).
Statistical analysis
Results were expressed as mean of three
individual experiments ± standard deviation
which was calculated using Microsoft excel.
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 278–291
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
RESULTS
Effect of ionizing radiation (IR) on cervical
cancer cells
For treatment with ionizing radiation HPV
positive cell lines were chosen i.e HPV 16
positive cell lines SiHa, CaSki which vary in
their HPV copy number. Cells were treated
with ionizing radiation doses ranging from 1Gy
to 3Gy, to assess their effect on these cell lines.
Gradual increase in apoptosis was found on
increasing radiation dose from 1Gy to 3Gy (Fig
1) but the sensitivity to ionizing radiation
varied from one cell line to another
demonstrating the variation of resistance to
radiation with cell type. Our data indicates that
Ionizing radiation dose of 1Gy results in 15.5%
apoptosis in SiHa and 18.5% apoptosis in
CaSki cells. Treatment with 2 Gy radiation
dose resulted in increase in apoptosis in both
the cell lines, in which SiHa showed 23.6% and
CaSki showed 19.08% apoptosis respectively,
hence demonstrating the radio-resistance.
Escalation of radiation dose to 3Gy resulted in
marginal increase in apoptosis 26.23% in SiHa
and 22.04% in CaSki cells. Since there was
only a marginal increase in apoptosis on
increasing radiation dose from 2Gy to the dose
of 3Gy, hence we settled for the radiation dose
of 2Gy for all the experiments. This dose was
used in combination treatments with cisplatin
and Berberine also.
Effect of Cisplatin and Cisplatin / IR on
cervical cancer cells
Platinum compounds like cisplatin in
combination with radiotherapy are used
extensively for treatment of cervical cancer so
we tried to mimic this situation in vitro on the
cell lines. First we assessed the effect of
cisplatin alone on these cell lines and in
combination with IR (2Gy). Treatment of SiHa
and CaSki with 5μg/ml cisplatin for 24 hrs
resulted in 19.4% and 12.95% apoptosis
respectively (Fig 2). Coupling this dose of
cisplatin with 2 Gy ionizing radiation (IR)
resulted in increase in apoptosis to 23.4% and
16.74% in SiHa and CaSki cells respectively.
Escalation of cisplatin dose to 10 µg/ml for 24
hrs resulted in increase in apoptosis in SiHa
and CaSki to 32.65% and 35.4%, whereas in
combination with 2Gy IR it increased to
35.25% and 38.42%. Thus a chemoradiation
dose of 10 μg/ml CP (cisplatin) / 2Gy IR was
selected for further experiments (Fig 2).
Effects of Berberine / CP / IR on cervical
cancer cells
Berberine has been established as a
potential chemotherapeutic agent in various
clinical trials and has been found to be well
tolerated at higher doses but its bio-availability
remains a major problem. Howell et al., (2007)
have summarized based on in vitro and in vivo
studies and clinical trials on natural products
like Curcumin and Berberine. It is shown that
the concentrations of natural products that were
achievable in the plasma of patients were only
at a lower micromolar range; hence, they have
suggested that for in vitro studies concentration
of berberine in less than 50 μM range do not
have any physiological relevance. The
significant radio-sensitization achieved by the
moderate dose of berberine at relevant doses in
vitro (2–6 Gy) has promising implications for
improving radiation therapy especially in radio
resistant tumors such as the tumors of the
uterine cervix. Hence we coupled the above
dose of 10 μg/ml CP with 50 μM berberine.
The cells were treated to this combination dose
for 24 hrs followed by 2Gy IR. An abrupt
increase in apoptosis was seen in the cell lines
with SiHa showing 49.8% apoptosis which was
higher than 35.25% apoptosis observed with
10 μg/ml CP/ 2Gy IR (Ionizing Radiation)
dose, used as standard therapy. Similarly,
CaSki showed an increase in apoptosis to
41.65% on treatment with the combination dose
against apoptosis achieved on treatment with
CP/ IR combination (Fig 2). Clearly our results
show that Berberine is selectively increasing
apoptosis in these cell lines. We then compared
the effect of Berberine/ CP combination with
CP/ IR, on various proteins involved in
apoptosis both pro-apoptotic and anti-apoptotic,
radio resistance and inflammatory response.
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 278–291
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
Fig 1
Microscopic and Flow cytometric analysis of apoptosis in SiHa (Fig 1A) and Ca Ski (Fig 1B) cells on treatment with IR
(1-3Gy).
Fig 2
Microscopic and Flow cytometric analysis of apoptosis in SiHa (Fig 2A) and Ca Ski (Fig 2B) cells on treatment with
CP/IR and 75µM Berberine/10µgm Cisplatin/IR for 24 hrs . The percentage Apoptosis shown in the bar diagram is
mean ± SD of three individual experiments
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 278–291
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
Effects of berberine / Cisplatin / IR and
Cisplatin / IR on protein p73
The protein p53 is a well-known tumor
suppressor protein that functions primarily as a
transcription factor, initiates cell cycle arrest
and apoptosis after genotoxic stress. Both the
cell lines used in the current study are HPV
(Human Papilloma Virus) positive cell lines
which encodes for viral protein E6 which
causes ubiquitin mediated degradation of p53
so another member of p53 family p73 becomes
important in these cells. It has been shown that
p73 plays an important role in Hydrogen
peroxide induced apoptosis (Singh et al., 2007;
Singh and Singh 2008). Since radiation induced
apoptosis involves ROS (Reactive oxygen
species) so we determined the effect of
cisplatin/IR and cisplatin/Berberine/IR
combination on p73 in these cell lines. In SiHa
and CaSki cells there was 36% and 44%
increase in p73 expression on treatment with
cisplatin/IR but on treatment with
Berberine/cisplatin/IR the expression of p73
increased to 54%, 98% (Fig.3B) demonstrating
that p73 level changes in response to Berberine.
Javvadi et al., 2008 have shown that ROS plays
an important role in Berberine/IR mediated
apoptosis in SiHa cells. Our results are in
agreement with their findings and suggest that
cisplatin/ Berberine /IR induced activation of
p73 which may involve reactive oxygen species
(ROS).
Effects of Berberine / Cisplatin / IR
combination on cyclin D1
Cyclin D1 is involved in cell-cycle arrest in
DNA-damage response. Cyclin D1 has been
shown to be induced by low-dose ionizing
radiation in human keratinocytes with an
adaptive radio-resistance (Ahmed et al., 2008).
On exposure of SiHa and CaSki to Cisplatin/IR
there was a 22% and 30% decrease in level of
cyclin D1 which was reduced to 49% in case of
CaSki but remained unchanged in SiHa on
treatment by Berberine/cisplatin/IR, thereby
demonstrating variation from cell type to cell
type (Fig.3A)
Fig 3
Effect on the level of Cyclin D1 (Fig 3A) protein and apoptotic p73 (Fig 3B) protein in SiHa and Ca Ski cells by western
blotting .Lane 1 control, Lane 2 CP 10μg for 24 hrs followed by 2 Gy IR, Lane 3 CP 10μg/75µM Berberine for 24 hrs
followed by 2Gy IR (Relative unit, RU). The results shown are mean ± SD of three individual experiments in the bar
diagram.
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 278–291
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
Berberine / Cisplatin / IR combination
treatment resulted in activation of
proapoptotic Bax and reduction in level of
Bcl XL
Cells on exposure to IR along with
chemotherapeutic agent results in DNA damage
and if this is severe, p53 and its counterparts
like p73 may trigger programmed cell death by
means of pro-apoptotic genes such as Bax and
inhibition of anti-apoptotic Bcl XL. It has been
demonstrated that radio resistant laryngeal
cancer was associated with increased Bcl-2 and
Bcl-XL expression and loss of Bax expression.
Bcl-2 family has been proposed to predict
radiotherapy outcome (Nix et al., 2005). The
association between expression of Bcl-2, Bcl-
XL and Bax with radio resistant cancer
suggests a potential mechanism by which
cancer cells avoid the destructive effects of
radiotherapy. We probed the effect of
Berberine/Cisplatin/IR combination on level of
Bax in SiHa and CaSki cells. The Bax
expression increased to 45% and 52% on
treatment with cisplatin/IR, but this increased
to 46% and 68% with respect to control on
treatment with Berberine/cisplatin/IR
combination.(Fig.4A) There was a marginal
increase in case of CaSki and nearly no change
in case of SiHa. However, in case of anti-
apoptotic Bcl XL we obtained a 7% decrease in
its level in case of SiHa and 15% decrease in its
level in case of CaSki cells on treatment with
cisplatin/IR (Fig.4B). In contrast, on treatment
with Berberine/cisplatin/IR we obtained a
substantial decrease in level of BclXL i.e. 22%
in SiHa and 26% in CaSki.
Berberine / Cisplatin / IR combination
treatment did not affect the expression of
COX 2 and AIF
COX-2 has been implicated in
carcinogenesis of systemic cancers. COX-2
inhibition has been shown to increase the radio-
sensitivity of various tumors. Results from the
present study demonstrate that on treatment of
SiHa and CaSki cells with cisplatin/IR there
was a 36% and 13% increase respectively in
expression of COX 2 but treatment of these
cells with Berberine/cisplatin/IR resulted in 8%
and 6% decrease respectively in level of
COX2(Fig.5A). These results indicate that
though Berberine/cisplatin/IR based therapy is
repressing COX 2 the effect is only marginal
and it varies with cell type. AIF expression was
unaltered remaining nearly the same on both
the treatments in both the cell lines (Fig 5B),
suggesting its noninvolvement.
Berberine / Cisplatin / IR treatment
enhances activation of both Caspase-3 and -9
Caspase -3 and -9 have been implicated to
play an important role in mitochondrial
mediated apoptosis by causing activation of
Caspase activated DNAse, finally causing
degradation of DNA. Hence we compared the
effect of Berberine/cisplatin/IR and cisplatin/IR
on Caspase -3 and -9 activities. The results
show that in SiHa and CaSki cells there was
1.19 and 1.23 fold increase in activity (Arbitary
fluroscence units, Afu converted to fold change
compared to control) in activity of Caspase -3
on treatment with cisplatin/IR, while there was
a 1.55 and 2.27 fold increase on treatment with
Berberine/cisplatin/IR (Fig 6A). Clearly
Berberine is causing activation of caspase -3.
To assess whether enhanced apoptosis was
being mediated through mitochondrial
pathway, we also studied the effect on caspase-
9 activity. SiHa and CaSki cells showed 0.29
and 0.11 fold increase in activity of Caspase -9
on treatment with cisplatin/IR while there was a
0.38 and 0.78 fold increase on treatment with
Berberine/cisplatin/IR. Thus Berberine appears
to be mediating apoptosis through
mitochondrial pathway (Fig 6B).
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 278–291
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
Fig 4
Effect on the level of antiapoptotic Bcl XL (Fig 4A) protein and apoptotic Bax (Fig 4B) protein in SiHa and CaSki cells
by western blotting .Lane 1 control, Lane 2 CP 10μg for 24 hrs followed by 2 Gy IR, Lane 3 CP 10μg/75µM Berberine
for 24 hrs followed by 2Gy IR (Relative unit, RU). The results shown are mean ± SD of three individual experiments in
the bar diagram.
Fig 5
Effect on the level of AIF (Apoptosis inducing factor) (Fig 5A) and inflammatory COX 2 (Fig 5B) protein in SiHa and
Ca Ski cells by western blotting .Lane 1 control, Lane 2 CP 10μg for 24 hrs followed by 2 Gy IR, Lane 3 CP
10μg/75µM Berberine for 24 hrs followed by 2Gy IR (Relative unit, RU). The results shown are mean ± SD of three
individual experiments in the bar diagram.
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Fig 6
Caspase -3 (Fig 7A) and -9 (Fig 7B) activity (Arbitrary fluorescence unit, Afu) after treatment with CP 10μg for 24 hrs
followed by 2 Gy IR and 10μg/75µM Berberine for 24 hrs followed by 2Gy IR in SiHa and Ca Ski. The results are mean
± SD of three individual experiments.
Fig 7
Effect of 10 μg CP for 24 hrs followed by 2 Gy IR and CP 10 μg/75µM Berberine for 24 hrs followed by 2Gy IR on
telomerase activity in SiHa and Ca Ski cells (RU). The results shown are mean ± SD of three individual experiments
Berberine / Cisplatin / IR combination
causes reduction in activity of Telomerase
with respect to Cisplatin/IR
Telomerase activation plays a critical role
in tumor growth and progression, in part by
maintenance of telomere structure. Indeed, the
ubiquitous expression of telomerase in human
cancers makes telomerase a promising target
for cancer therapy. We assessed the effect of
Berberine/cisplatin/IR combination dose on
telomerase with respect to cisplatin/IR
combination dose by using PCR/ELISA
methodology which assesses activity of
hTERT. We observed that on treatment of SiHa
and CaSki there was 22% and 39% decrease in
telomerase with cisplatin/IR while there was an
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82% and 75% decrease in telomerase activity
on treatment with Berberine/cisplatin/IR
(Figure 7). This data suggests that berberine
based therapy causes substantial decrease in
telomerase activity and this therapy can be a
potent telomerase targeted approach for
treatment of cervical cancer.
DISCUSSION
Cervical cancer remains one of the major
killers amongst women worldwide. Chemo/
radiotherapy regime which is cisplatin based
radiotherapy is used for the treatment of
advanced cervical cancer in India. Evidence
shows that most of the chemotherapeutic drugs
used in current clinical practice are radio-
sensitizers. Several newer cytotoxic agents with
radio-sensitizing properties are being tried in
combination with cisplatin but their use is
generally limited by dose related toxicities
(Rosa et al., 2012). Natural products open a all
new avenue for treatment of cancer as they are
generally tolerated at high doses. Animal
studies have confirmed the anti-tumorigenic
activity of natural products like Berberine and
curcumin (Howells et al., 2007). Phase I
clinical trials on curcumin showed that it is safe
to humans up to 12,000 mg/day when taken
orally (Cheng et al., 2001; Sharma et al., 2001;
Lao et al., 2006) and caused histological
improvement of precancerous lesions in
patients, suggesting that it is biologically active
at these doses (Cheng et al., 2001). Previous
reports have indicated that Berberine confers
radio-sensitizing effects in prostate and
squamous cell carcinoma cell lines, and
recently in cervical cancer cell lines HeLa and
SiHa. Thus we evaluated the effect of
Berberine in combination with cisplatin/IR and
compared it with cisplatin/IR treatment. Our
results suggest that Berberine/cisplatin/IR
based therapy is likely to be more effective and
safer to treat cervical cancer, as Berberine is
well tolerated in humans, even at high doses.
We present in vitro evidence that this approach
targets radio-resistance and anti-apoptotic
proteins in a much more specific way compared
to the standard cisplatin/IR based therapy.
When cells are exposed to clinically
relevant doses of ionizing radiation it causes
DNA damage by generation of reactive oxygen
species (ROS). This DNA damage causes a
rapid ROS dependent activation of pro-
apoptotic and anti-apoptotic cascade which
involves activation of p53 family of genes
acting downstream to trigger apoptosis if the
damage is not repairable. The tumor cells are
dynamic with respect to their reliance on
specific cell signaling pathway to exist and
rapidly adapt to repeated toxic challenges in an
attempt to maintain tumor survival. Prolonged
inhibition of any one of these pathways
however, gives rise to lineage of cells which
become resistant to inhibitor drug, with point
mutations in the specific targeted proteins, or
by reprogramming of multiple signaling
processes within the cell (Valerie et al., 2007).
So the need for today is therapies which target
these multiple pathways and result in selective
apoptosis of cancer cells and thus deal with the
problem of radio-resistance. The cell lines used
in this study are HPV positive, in which wild
type p53 is targeted for ubiquitin mediated
degradation so other members of p53 family
like p73 become particularly important in these
cell lines (Singh et al., 2007; 11 Singh and
Singh 2008). We observed the activation of p73
in cisplatin/IR treated sample but this activation
was much more in berberine/cisplatin/IR
treated samples which indicates that
combination of Berberine is causing much
more activation of p73 probably through
mediation of ROS. This has been demonstrated
recently in some studies in which natural
products like Berberine were shown to be a
pro-oxidant in combination with ionizing
radiation, and its radio-sensitizing properties
were attributed to ROS mediated signaling
(Javvadi et al., 2008).
Bax and Bcl XL are members of Bcl 2
family of proteins which control apoptosis and
are associated with regulating the
mitochondrial membrane permeability. Pro-
apoptotic proteins like Bax by translocation
from the cytosol to the mitochondria, induce
cytochrome c release, whereas Bcl-XL exerts
its anti-apoptotic activity, at least in part by
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 278–291
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inhibiting the translocation of Bax to the
mitochondria (Mohammad et al., 2008; Ow et
al., 2008). Our results show a slight increase in
Bax and a decrease in Bcl XL on Berberine
treatment in both the cell lines. AIF (Apoptosis
inducing factor) remained unchanged thereby
suggesting its non involvement.
Caspases, a family of aspartic acid-specific
proteases, are the major effectors of apoptosis.
Once activated, caspases preside over the
ordered dismantling of the cell through
restricted proteolysis of hundreds of substrate
proteins (Ow et al., 2008). Caspase -3 has been
implicated in both the extrinsic and intrinsic
pathway of apoptosis. Our findings show
activation of caspase -3 on both the treatments
but this activation was more marked on
Berberine/cisplatin/IR treatment implicating
that cells are undergoing apoptosis through
caspase-3 mediated pathway. Similarly,
caspase -9 was activated on both the treatments
in both the cell lines, but was more marked on
Berberine/cisplatin/IR treatment, implicating
involvement of mitochondrial mediated
apoptosis pathway. Next we assayed the effect
of Berberine/cisplatin/IR on proteins involved
in radioresistance like Telomerase, Cyclin D1
and Cox 2.
Cyclin D1 is involved in cell-cycle arrest in
DNA-damage response. Cyclin D1 contributes
to regulate G1 progression by forming a
complex with different cyclin-dependent
kinases. It has oncogenic properties and is
frequently overexpressed in several human
tumor types. Cyclin D1 has been shown to be
induced by low-dose ionizing radiation in
human keratinocytes with an adaptive radio-
resistance (Ahmed et al., 2008). Our results
show a similar reduction in level of cyclin D1
on berberine treatment in both cell lines, but to
different extents, demonstrating cell to cell type
variation.
Cyclooxygenase-2 (COX-2), an enzyme
induced by pro-inflammatory cytokines,
mitogenic substances, oncogenes, growth
factors, and hypoxia, among others, is involved
in the metabolic conversion of arachidonic acid
to prostaglandins in inflamed tissues and
neoplasia. COX-2 is often overexpressed in
malignant tumors and premalignant lesions.
Because COX-2 may also be a determinant of
tumor radio-resistance, its inhibition or
inhibition of its products (prostaglandins) may
improve tumor response to radiotherapy.
Analyses of the effect of cisplatin/IR show that
COX 2 was elevated in both the cells on
treatment with cisplatin/IR indicating that this
radio-resistant marker is elevated in all the cell
lines however on treatment with berberine
cisplatin/IR COX 2 was down-regulated in both
the cell lines.
Telomeres and telomerase play a role in
the regulation of the life span of the cell.
Human cells express low levels of telomerase;
however when telomere length reaches a
critical level, abnormal activation of telomerase
can lead to immortalization and uncontrolled
proliferation (Agarwal et al., 2008; Gandellini
et al., 2007). Our result shows that Berberine
treatment caused a substantial reduction in
telomerase activity in SiHa and CaSki cervical
carcinoma cell lines. Thus Berberine is
conferring a selective advantage over cisplatin
based radiotherapy by causing marked decrease
in telomerase activity.
In summary, our data provides in vitro
evidence that supports the clinical importance
of coupling Berberine with cisplatin as an
efficient radiosenitizer for treatment of cervical
cancer as it causes enhanced activation of p73
causing further down-regulation of anti-
apoptotic Bcl XL and activation of caspase -3
and -9, resulting in enhanced apoptosis. We
also provide evidence that Berberine/cisplatin
based radiotherapy causes substantial down-
regulation of telomerase activity, Cyclin D1
and COX 2, thus acting as a potent radio-
sensitizer. This data has clinical relevance as it
highlights the importance of overcoming the
problem of radio-resistance by specifically
targeting radio-resistant proteins using
Berberine which is well tolerated in human
beings even at high doses, and thus promises to
be effective drug in future clinical trials along
with cisplatin/IR.
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 278–291
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
ACKNOWLEDGEMENT
We would like to acknowledge DBT for
financial support to Komal. This work was also
supported by SRF grant to Mayank Singh from
CSIR. We also acknowledge the technical help
from Jyotibasu for carrying out IR Treatment at
IRCH, AIIMS.
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Source of Support: Nil Conflict of Interest: None Declared
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ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
PHYTOCHEMICAL SCREENING OF SECONDARY METABOLITES OF
EUPHORBIA NERIIFOLIA LINN.
Chouhan Leela¹, Bhatt Shashank²*, Dhyani Suresh3
1Department of Biotechnology, Rajiv Gandhi Govt. P.G. College, Mandsaur, Madhya Pradesh, 458001,
India. 2,3
Department of Biotechnology, NIMS University, Jaipur, Rajasthan, 303121, India. *Corresponding author: Email: [email protected]; Mobile: +919826840428
Received: 02/04/2013; Revised: 25/04/2013; Accepted: 02/05/2013
ABSTRACT
Metabolites play an important role for the protection of plants against various diseases. Each
secondary metabolite has a specific activity. Euphorbia neriifolia is one such medicinal plant with
maximum secondary metabolites. These metabolites were extracted from different polar solvents -
petroleum ether, chloroform, methanol, 95% ethanol and distilled water. Alkaloids, flavonoids,
glycosides, phenols, saponins, sterols, lignins, and tannins were found in Euphorbia neriifolia’s
flowers with cold percolation method. Each metabolites show effective activities against diseases.
KEY WORDS: Alkaloids, flavonoids, sterols, Euphorbia neriifolia Linn.
Research article
Cite this article:
Chouhan Leela, Bhatt Shashank, Dhyani Suresh (2013), PHYTOCHEMICAL SCREENING OF
SECONDARY METABOLITES OF EUPHORBIA NERIIFOLIA LINN., Global J Res. Med. Plants &
Indigen. Med., Volume 2(5): 292–297
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 292–297
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
INTRODUCTION
Plants have different types of therapeutic
properties by which different ailments to
mankind are dealt with. Medicinal plants
naturally accumulate different types of primary
and secondary metabolites. Different types of
medicinal plants have been used for the
treatment of diseases. The quantity of
secondary metabolites vary in different parts of
a plant and is also influenced by environmental
conditions (Aanchal Jain et al., 2012; Surendra
K. Rathore et al., 2012; Priyanka Parmar et al.,
2012).
Plant species are widely used in Indian
systems of medicine since times immemorial
and one such ancient plant is Euphorbia
neriifolia Linn. The general name of Euphorbia
neriifolia Linn. is Sehund /Thohar in Hindi and
Dog’s Tongue in English. It is known as Indian
spurge tree, Oleander spurge, Hedge
Euphorbia, Snuhi (Sanskrit) and Ilachevikalli
(siddha). This plant belongs to the family of
Euphorbiaceae (Burkill IH; 1936, Ambasta SP;
1986).
Generally Euphorbia neriifolia grows
luxuriously around dry, rocky, North hilly
areas, Central and Southern parts of India. The
leaves are thick, greenish, 6–12 inches long and
ovular shape (Anonymous, 1952). Most of the
people know about it that it is very toxic and
waste plant but recent researches have shown
that it has medicinal uses in the treatment of
typical diseases. It is a bitter laxative,
carminative and useful in abdominal troubles,
bronchitis, tumors, leucoderma, piles,
enlargement of Spleen, anemia, ulcers, fever
and in chronic respiratory troubles (AK
Nadkarni, 1954).
MATERIAL AND METHODS
Collection of Plant Material
Euphorbia neriifolia Linn. is found all over
India. The plant flowers were collected from
Mandsaur district, Madhya Pradesh. Mandsaur
District forms the northern projection of
Madhya Pradesh (Fig.-1). It lies between the
parallels of latitude 23° 45' 50" North and 25°
2' 55" North, and between the meridians of
longitude 74° 42' 30" East and 75° 50' 20" East.
Fig.1- Map of Madhya Pradesh
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 292–297
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Fig.2- Euphorbia neriifolia Linn.
Preliminary Screening of Secondary
Metabolites
The flower of Euphorbia neriifolia (Fig.2)
were collected from Mandsaur Madhya
Pradesh.The plant was identified to Dr. S.N.
Mishra, Principal Scientist, All India
Coordinated Research Project on Medicinal and
Aromatic Plants, College of Horticulture,
Mandsaur, affiliated to R.V.S.K.V.
Vishwavidyalaya, Gwalior.
The flowers were kept in shade, dried and
powdered using mixer grinder, and subjected to
cold percolation process for 48 hours with
petroleum ether, chloroform, methanol, 95%
ethanol and distilled water. After this process,
the extracts were filtered and used for
preliminary phytochemical screening such as
alkaloids (Iodine, Wagner, and Dragendorff’s
test), flavonoids (Pew’s, Shinoda and NaOH
tests), glycosides (Keller-Killani, Conc. H2SO4,
and Molisch tests), lignin (Labat and Lignin
tests), phenols (Ellagic acid and Phenol tests),
saponins (Foam and Haemolysis test), sterols
(Libermann- Burchard, and Salkowski tests),
tannins (Gelatin and Lead acetate tests) were
carried out (Shashank Bhatt et al.,2011).
Preliminary Screening of Phytochemical
Test
Phytochemical Screening
The filtrate obtained was subjected to
preliminary phytochemical screening.
For identification of alkaloids Iodine test
(Khandelwal K.R., 2008), Wagner’s test
(Kokate C.K. et al., 2001) and Dragendorff’s
test (Kokate C.K. et al.,2001) were performed.
For identification of flavonoids Pew’s test
(Peach K., Tracey MV. 1956), Shinoda test
(Kokate C.K. et al., 2001) and NaOH test
(Khandelwal K.R., 2008) were performed.
For identification of glycosides Keller-Killani
test (Kokate C.K. et al., 2001), Glycoside test
(Treare GE, Evans WC. 1985), Conc. H2SO4
test (Khandewal K.R., 2008) and Molish test
(Kokate C. K. et. al., 2001) were performed.
For identification of Phenol Ellagic acid test
(Gibbs R.D., 1974) and Phenol test (Gibbs
R.D., 1974) were done.
For identification of lignin lignin test (Gibbs
R.D., 1974) and labat test (Gibbs R.D., 1974)
were performed.
For identification of saponins foam test
(Kokate C. K. et. al., 2001), and Haemolysis
test (Kokate C.K., 1994) were done.
For Identification of sterols Liebermann-
Burchard test (Kokate C. K. et. al., 2001) and
Salkowski’s test (Kokate C. K. et. al., 2001)
were done.
For identification of tannins gelatin test (Treare
GE, Evans WC. 1985) and lead acetate test
(Treare GE, Evans WC. 1985) were performed.
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 292–297
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TABLE-1: Phytochemical Screening of Euphorbia neriifolia Linn. Flowers
Test Petroleum
Ether
Chloroform Methanol 95% Ethanol Distilled water
Alkaloids
Iodine Test − − − − −
Wagners Test − − + + −
Dragendorff’s
Test
− − + + −
Flavonoids
Pews Test − − + + −
Shinoda Test − − + + −
NaOH Test − − + + −
Glycosides
Keller- Killani
Test
+ + + + +
Glycosides Test − + + + −
Conc. H2SO4 + + + + +
Molishs Test + + + + +
Phenol
Ellagic Test − − + + −
Phenol Test − − + + −
Lignin
Lignin Test − + + + −
Labat Test − + + + −
Saponins
Foam Test − − + + +
Haemolysis Test − − + + −
Sterols
Libermann-
Burchard Test
+ + + + +
Salkowski Test − + + + −
Tannins
Gelatin Test − − + + −
Lead Acetate Test − − + + +
(+)Presence, (−) Absent
RESULT AND DISCUSSION
The results of the phytochemical screening
of Euphorbia neriifolia’s flowers’ have been
presented in Table-1. It showed that different
types of secondary metabolites such as
alkaloids, flavonoids, glycosides, phenol,
lignins, saponins, sterols and tannins were
present. Most of the secondary metabolites
present in flowers show the good therapeutic
activity against diseases compared to other
parts. According to the guidelines of WHO &
Ayurveda, if a plant has anti-pathogenicity,
anti-cancer and different types of activities it
can be involved into medicinal plant category.
Secondary metabolites are these types of
compounds.
Flavonoids have inherent abilities to
modify the body’s reaction to allergen, virus
and carcinogens. Tannins have general
antimicrobial and antioxidant activities
(Rievere et al., 2009). Current reports show
that tannins may have potential values such as
cytotoxic and antineoplastic agents (Aguinaldo
et al., 2005). Plant steroids have cardiotonic
activity, possess insecticidal and antimicrobial
properties. It is generally used in herbal
medicines and cosmetic products (Callow;
1936).
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 292–297
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CONCLUSION
Euphorbia neriifolia Linn. has different
types of medicinal properties. It has different
types of secondary metabolites that have been
presented in phyto-chemical screening of
secondary metabolites study. These secondary
metabolites have anti-HIV, anti-bacterial, anti-
viral, anti-fever, anti-diabetic, anti-cancerous
activities etc. Therefore, it can be involved in
medicinal plant category.
ACKNOWLEDGEMENT
The authors are very thankful to Dr. S.N.
Mishra for assist in identification of plant and
kind assistance of my dear father Shri Krishna
Kumar Bhatt, Bharti, Smt. Subhadra bhatt and
brother Mr. Mayank Bhatt. Last but not least
the Almighty God is unforgettable without
whose kindness and grace, anything was not
possible.
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Source of Support: Nil Conflict of Interest: None Declared
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ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
TRADITIONAL MEDICINES OF HERBAL ORIGIN PRACTICE BY THE
ADI TRIBE OF EAST SIANG DISTRICT OF ARUNACHAL PRADESH, INDIA
Das Moushumi1*, Jaishi Anju
2, Sarma Hirendra N
3
1Assistant Professor, Department of Zoology, University of Science and Technology, Meghalaya, Ri-Bhoi
District, Meghalaya, India 2Research Scholar, Department of Zoology (Center with Potential for Excellence in Biodiversity), Rajiv Gandhi
University, Itanagar – 791 112, India 3Professor, Department of Zoology (Center with Potential for Excellence in Biodiversity), Rajiv Gandhi
University Itanagar – 791 112, India *Corresponding Author: Email: [email protected]; Mob: +919508419900
Received: 01/03/2013; Revised: 17/04/2013; Accepted: 25/04/2013
ABSTRACT
A survey was conducted on the use of traditional medicine by the Adi tribe of East Siang district
of Arunachal Pradesh, a state of eastern Indian Himalayan Mountain range. Ten numbers of
Traditional Medicine Practitioners (TMP) belonging to the age group of 45–60 years were interacted.
Data were collected preparing semi structured questionnaire. Thirty three plant species were
identified as being used for Traditional Medicine preparation by the Adi people. Eighty one percent
of these plants were used for human, while 19% were used for animal diseases. Traditional medicines
were used for treatment of Jaundice, typhoid and malaria (9%), eye infection (6%) burn injuries
(6%), skin disease and repellant (6%), diarrhea (9%), toothache (9%), wound healing and infection
(15%), bone fracture (15%) and fertility control (24%). Some of these TM were prepared in a
composite form using multiple number of plants or part thereof. Use of TM is still prevailing among
the villagers as one of the cheapest and easily available medical practice. It has been speculated that
proper screening of these herbal medicines shall provide new insight on the hitherto unexplored
traditional medicines of Adi people of Arunachal Pradesh.
KEYWORDS: Abortifacient, Antifertility, Traditional medicine, Indigenous Knowledge
Research article
Cite this article:
Das M, Jaishi A, Sarma H N (2013), TRADITIONAL MEDICINES OF HERBAL ORIGIN PRACTICE
BY THE ADI TRIBE OF EAST SIANG DISTRICT OF ARUNACHAL PRADESH, INDIA,
Global J Res. Med. Plants & Indigen. Med., Volume 2(5): 298–310
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INTRODUCTION:
Arunachal Pradesh a state of the Eastern
Himalayan region of India is one of the Mega
Biodiversity hot spots of the world. This
Biodiversity hot spot zone is characterized by
wide variety of floral and faunal population.
Along with a variety of plants and animals this
state is enriched with a number of ethnic
groups of people. These people have an age-old
history and practice of using Traditional
Medicine (TM) of herbal origin for treatment of
various ailments/ conditions of both human and
animals. Arunachal Pradesh is sharing
international border with China and Myanmar.
Thus, it was earlier known as North East
Frontier Agency (NEFA) and governed by
Government of India as Union Territory till
20th
February 1987. The NEFA gets the
statehood on 20th
February, 1987 with its new
identity of Arunachal Pradesh (http://
arunachalpradesh.nic/people.htm.). Belonging
to the eastern Himalayan Biodiversity rich
zone, this hill state is the home land of more
than 25 tribal people groups
(http://WWW.indianetzone.com/8/tribes_aruna
chal_pradesh.htm). The „Adi‟ is one of the
major tribe among these groups of tribal
population of Arunachal Pradesh. The name
„Adi‟ was earlier known as „Abor‟, (Dalton
E.T, 1872). The transformation of the name
from „Abor‟ to „Adi‟ took place after the
development of disregard to the meaning of
„Abor‟. The use of the word „Abor‟ which
means „unruly‟ or „savage‟ has been traced
back to the first century A.D., that a great
valley of Himalayas called „Aborimon‟ was
inhabited by wild men (Hutton J.H, 1946).
However, the term „Abor‟ has been used with a
broader sense of meaning of „independence,
unruly, savage‟ and so on and applied to all the
hill tribes of valley of Assam (Mackenzie A,
1884). The original home of Adi people has not
yet been studied and ascertained. According to
some authors Adis are considered to be origin
in the lofty mountain ranges of Tibet and
Assam (Nyori Tai, 1993). Coming from the
northern and north-eastern part of greater
Himalayas, these Adi people moved
southwards and settled in the erstwhile „Siang
frontier division‟ of NEFA. This „Siang
Frontier Division‟ spreads from Subansiri
River in the west and Dibang River in the east.
With advancement of time the Siang Frontier
Division has been demarcated into three Siang
districts (East Siang, West Siang and Upper
Siang) and Dibang valley district of present day
Arunachal Pradesh. Damroh, a Padam village
in the East Siang district has been considered as
the epicenter of the dispersal of the Adi people
(Nyori Tai, 1993). In the course of time, Adis
are divided into different groups, such as
Padam, Minyong, Pangi, Shimong, Ashing,
Pasi, Karko, Bokar, Bori, Ramo, Pailibo,
Milan, Tangam and Tagin (Roy S 1966). The
Adis belong to the Mongoloid stock and thus
similar to those of the other of the same stock
(Chatterjee S.K, 1951). The history of the Adi
people‟s relationship to modern world dates
back to early 19th
century; when Captain
Bedford, the first European from Great Britain
visited the Adi inhabited area of Eastern
Himalayas in the year 1825–26. In the early
part of 20th century, the Adi villages came
under the British authority (Roy S, 1966).The
traditional Adi villages are organized under a
village council called „Kebang‟ in Adi
language (Rizvi et al., 2006). The „Kebang‟ is a
powerful body honored by all sections of Adi
people and involved in looking after over all
social development as well as settlement of
social dispute amicably. The “Adi language” is
the lingua franka of the Adi people. These
indigenous people are self-sufficient in most of
the spheres of life in their remote villages. They
collect their food and medicinal requirements
from the untouched natural resources of
neighboring forest. In the present investigation
such plants and part there off used for cure of
various ailments like diarrhea, fever, jaundice
etc., treatment for bone fracture and for
reproduction control especially of domesticated
animals practiced by the Adi tribe of East Siang
district have been documented through first
hand information collection. The quest for
development of novel drugs using herbs
mentioned in various folk literatures is in force
in the scientific community. In such herbal
preparation plants are used either singly (parts
thereof) or in composite form. It has been
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speculated that the present study shall be
helpful for preservation of indigenous
knowledge on TM, its preparation and
application, over & above the societal
development of these tribal communities in
India.
MATERIALS AND METHODS
Study area
The East Siang district is located in
between 27°43´ and 29°20´ North latitude and
94°42´ and 95°35´ East latitude (Fig.1). The
district has a total of area of 4005 sq.km
covering the other bank of Siang River. It
spreads over a wide range of altitudinal height
of mean sea level from a minimum of 133 ft
(Ruksin area) and maximum 752 ft (Riga).
Siang River flows from northern part of lofty
mountains of Arunachal Pradesh and merges
with the Brahmaputra River in plains of Assam.
The „Siyom‟ and „Siku‟ are two main
tributaries of Siang River, flowing across the
west and east bank of the mighty Siang
respectively. Thus, the Siang River flows
throughout the entire district creating the Siang
valley in Arunachal Pradesh. The entire district
is characterized by presence of series of spurs
of mountains. The outer range of the mountain
at the southern part of the district touches the
northern part of the Brahmaputra basin
bordering Assam. The northern mountain range
of the district is densely covered with forest.
The southern portion is comparatively flat and
densely wooded except the places having
human settlement. The climate of the district is
tropical humid in summer and sub temperate
during winter. It receives heavy rainfall every
year during monsoon season starting from May
till October. A cold dry seasonal wind blows
throughout the winter from November to
February. The soil texture varies from sandy to
loamy sand. Cultivation and rearing of
livestock is the main way of livelihood for
majority of the population. The dominant crop
of Adi people is the paddy followed by maize,
millet, ginger, mustard, potato, orange,
pineapple etc grown on the hill slopes.
Fig 1. Study Area: The East Siang District of AP
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Fig 2. Adi Female (A) and Adi male (B) Traditional Medicinal Practitioners (C) and an Adi
village (D).
Data collection
The data on medicinal use of plants practiced by the „Adi‟ peoples were collected primarily from rural villages of East Siang district. The villages namely Bogo Bango, Mikung, Ngorlung, Ledum, GTC Pashighat, Oyan, Mebo, Boleng and Pangin were selected for information collection. Villages were selected with a view to collect the information covering different circles of East Siang district. A „circle‟ consists of two or more villages within the district. Very often the names of the villages are found to be similar with the name of the circle it belongs to, e.g „Mebo‟ village is situated in the „Mebo‟ circle. Many of the villages are located in the remote areas accessible only on foot. During the survey, it was observed that a good number of aged people (both male and female folk) possess knowledge on traditional medicines. Among these villagers, the authors met the Traditional Medicine Practitioner (TMPs) who regularly prepare the TM and do practice and prescribe among the villagers. These people (TMP) belonging to age group of 40–65 years are locally known as „Miri‟. During the present investigation 10 numbers of „Miri‟s both male and female (Fig-2) were interacted to obtain information on the use of medicinal plants. In addition, 29 local informants of the villages were approached for information on the herbal medicines. The local informants neither practice nor prescribe the TM among the villagers, however, use this knowledge and
information for their own requirement whenever necessary.
The authors selected the „Miris‟ and local informants by approaching and interacting with the village heads, locally called as Gaon Burha of the surveyed villages. The Gaon Burha is the head of the village administration. The Gaon Burha or village head can take decisions/ order to settle any minor disputes within the The Gaon Burha also got the knowledge about the various activities going in and around the village. Some of the informants and „Miris‟ neither understands nor can speak the language other than „Adi language‟, In such situations local Adi people knowing both „Adi‟ and English languages were used as interpreters and mediators for data collection. Important to mention that many Adi people with higher education have been working as TMP and pursue the practice of TM among the villagers in East Siang district. Survey was carried out ten times in a time span of 23 months starting from the month of January 2005. The ethno medicinal data was collected following standard methods (Martin GJ., 1995; Jain SK., 1964) through general conversation with the informants, using semi structured questionnaire, interviews and the participants‟ observations.
The Questionnaire used for data collection 1. Name of the person 2. Sex 3. Age 4. Locality 5. Utilization of the plant for medicine
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(yes/No) 6. Plant medicine use for fever/bone fracture/sterility/abortion/other conditions 7. Description of plant 8. Mode of preparation/ parts used/composition of medicine 9. How much it is effective (temporary or permanent) 10. Mode of application/Procedure (eg., Dose, duration, time etc.) 11. Side effects, if any (whether vomiting, Nausea, weakness or any other adverse effect)
The Adi people have deep belief on the Sun and the Moon locally termed as “Donyi Polo” in Adi language and thus developed their own religion called “Donyi- Poloism”. Very often, preparation and practice of the traditional medicine is associated with worships and superstitions among these people. The data originated from the epistemology of indigenous knowledge on traditional medicines were reviewed and evaluated within epistemology arising from scientific reasoning and knowledge. The TMPs were interviewed with a view to exploring (1) sources of medicine, preparation and prescription, (2) TM associated with traditional belief associated with worships, (3) route of administration and application, (4) method of preparation, (5) Part used, (6) used for human and/ or animal. The authors accompanied the TMPs with the local interpreters to the field for observation and collection of plant species used for TM preparation. Plant species shown by the local practitioner were collected and stored in the form of herbarium in for identification.
Collection and preservation of the plant species were made following standard methods (Jain et al., 1976). Identification of the plant species was done using reference herbarium materials, various volumes of books on Indian medicinal plants (Kirtikar K.R et al., 1935; Hooker JD.1872–1897) and by taxonomist from Rajiv Gandhi University in Itanagar, Arunachal Pradesh.
RESULTS
The present study recorded a total of 33 plant species belong to 25 families (Table-1)
used as components of TM of Adi people. These herbal preparations were used for treatment of both animal and human being. Nineteen percent (19%) of the presently recorded plants have been used for treatment of animal diseases while remaining 81% are used for treatment of human diseases (Fig-4). Results showed that these TM are used for treatment of Jaundice, typhoid and malaria (9%), eye infection (6%) burn injuries (6%), skin disease and repellant (6%), diarrhea (9%), toothache (9%), wound healing and infection (15%), and bone fracture (15%). Twenty four percent (24%) of plants documented were used for preparation of TM for fertility control of both human and domestic animal (Fig-3). Mode of use of the plant parts varied according to the medicines prepared and its uses. Application of the fresh leaves was found to be the highest (51%) used among the collected information. 33% medicines were prepared from stems. Roots, tubers, rhizomes were used for preparing 18% of the traditional medicines recorded so far among the Adi people. Similarly, barks (9%), fruits and seeds (6%) were used for preparation of various traditional medicines by the Adi people. The medicines were prepared for both external (topical) application and through oral route depending upon the use of the medicines for ailments. Paste of freshly collected leaves were used for external application in wound healing and infection covering 52% of the application of the TM. Dry powder of the plants or parts thereof was used in 15% of the medicines taken along with food or water for treatment of diseases like malaria, diarrhea, jaundice, typhoid etc and for fertility control of animal and human. The crude juice and/ or sap and boiled residues of freshly collected plants (leaves, stems) were used in 33 % of medicines collected during the present investigation. While most of the medicinal plants were reported to be used in freshly collected form, some reported to be used in dried form. Plants belong to families Apocynaceae, Poaceae, Rutaceae, Rubiaceae, Scrophularaceae and Mimosaceae were reported to use for more than one TM preparation. The collected plant parts were stored in the museum of Life science Department, Rajiv Gandhi University, Itanagar.
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Table -1. Plants and its medicinal use prevail among the Adi people of East Siang district
Name of the plant (Family)&
No./vernacular name Part
used Use
Mode of preparation/use
Alstonia scholaris (L.) Br
(Apocynaceae) Adi/RGU/G-01/ Cingar
Leaves Wound healing,
malaria,
abortifacient in
pigs.
Paste of fresh leaves. Paste is mixed
with the meal of cattle to feed for
wound healing and same preparation
use in pigs for abortion.
Amphineuron extensus
(BI.)HOLTT (Thelypteridaceae) Adi/ RGU/G-02/ Rokji
Leaves Tick repellent in
chicken
Fresh leaves collected from the plant
are kept inside the poultry farm to
repel and/ or removes the tick from
the animal.
Ageratum conyzoidesL
(Asteraceae) Adi/ RGU/G-03/ Namsing
Leaves
Wound healing Paste is prepared from fresh leaves.
The paste is applied in the cut or
wound to stop bleeding and healing
of the wound. Doses depend on the
intensity of the wound.
Axonopus compressus
(SW)P.Beauv (Poaceae) Adi/ RGU/G-04/Bobosa
Roots Eye infection/
Disease Root is chewed by the patient to get
relief. Three times for three days.
Bannaya reptans Sprengel
(Scrophulariceae) Adi/RGU/G-05/Kat-Buk usueng
Leaves Nail infection
Freshly collected leaves are warmed
to make a paste. The paste is wrapped
with clean cloth at the site of
infection. Generally it is used at
night during sleeping hours.
Bryophyllum pinnatum. Kuntz.
(Crassulaceae.) Adi/ RGU/G-06/ Nebi – Nelum
Leaves Skin burn,
diarrhea, fracture
and Sciatica.
Pastes of fresh leaves are applied to
burnt area and use for fracture. For diarrhea,2–3 fresh leaves are
recommended to eat 4–5 times ina
day (for 4 days)
Clacaria macrophylla Wall
(Rubiaceae.) Adi/ RGU/G-07/ Pemi lagin
Sap of
the stem Conjunctivitis,
Eye infection
1–2 drops of the sap of the stem is
poured into the eye during infection
Clerodendrum spp (Nerbenaceae) Adi/ RGU/G-08/ Dumkar
Fresh
leaves Skin Burn Paste is prepared from fresh leaves to
apply on the burnt area.
Coptis teeta ( Ranunculaceae ) Adi/ RGU/G-09/Mishimi teeta
Roots,
leaves Malaria, Jaundice,
Typhoid and in
fertility control in
women
(abortifacient)
Powder is prepared from shade dried
Roots of Coptis teeta. The powder is
mixed with water and the mixture is
boiled for few minute to feed the
Malaria patient. For Abortive use
Preparation is made by mixing the
leaves with Caraca papaya, Moringa
oleifera (bark),Solanum spirale
(roots) and bark of Alstonia
schlolaris.
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Drymaria cordata Roem and
Schuletes (Caryophyllaceae) Adi/
RGU/G-10/ Taggom
Leaves
and stem Skin disease and
Bone fracture
Paste is prepared from the tender
branches of the plant. Applied on the
fractured area skin.
Laportia crenulata Gaud
(Urticaceae) Adi/RGU/G11/Matpepereng
Leaves
and stem Wound infection A paste is prepared from freshly
collected leaves and applied on the
wound of animals.
Melothria leucocarpa (BP) Cong
(Cucurbitaceae) Adi/ RGU/G-12/Pumroll
Leaves Wound infection
in animal Paste of Fresh leaves is mixed with
the meal of domestic animals.
Mikenia mirantha L. (Asteraceae) Adi/RGU/G13/Arunachal Eng
Leaves Cut, wound
healing Freshly collected leaves are grinded
to make paste and applied on wound/
cuts to heal the wound and for
prevent bleeding.
Oroxylum indicum L. Vent.
(Bignoniaceae) Adi/ RGU/G-14/Domir ettkung
Bark Tonic for malaria
patients Small pieces bark are sun dried and
then boiled with leaves of Ocimum
sanctum, Zingiber zerampet
(Rhizomes) and Mentha spicata. The
boiled residue is advised to use
during the whole treatment period.
Psidium guajava L. (Myrtaceae) Adi/ RGU/G-15/ Mudhuri anne
Leaves
/buds
Treatment of
diarrhea. Fresh tender leaves are advised to
chew by the person suffering from
diarrhea. Doses are 4–5 fresh twigs
takes twice or thrice a day for 3–4
days.
Paederia foetida (Rubiaceae.) Adi/RGU/G-16/Yepetare
Leaves Stomach disorder/
Diarrhea Fresh leaves are boiled with other
vegetables. The boiled residue is
given to eat to the patient to cure
fever and diarrhea. The leaves are
tied on head or hangs on neck to cure
headache and sinusitis.
Solanum nigrum L (Solanaceae)
Adi/ RGU/G-17/ Bangko
Leaves,
roots and
stem
Diarrhea, Malaria
and conjunctivitis Paste prepared from leaves is boiled
in water. Diarrhea and malaria
patient‟s treated with both the boiled
leaves paste and the extract for 4–5
days. The grinded roots are given to
the malaria patient to get rid of the
parasite. Mildly heated leaves are put
on the eye to cures conjunctivitis.
Zanthoxylum acanthopodium DC
(Rutaceae) Adi/ RGU/G-18/ Ombe
Stem
Tooth ach
The stem of plant is used. Chewing
or brush with it gives immediate
relief from the toothache and to
makes the teeth strong.
Zanthoxylum hamiltoniamun wall
(Rutaceae) Adi/ RGU/G-19/ Ongar
Leave,
fruit Tooth ache,
Abortifacient Leaves are dried on the sunlight or
smoke and then applied on the tooth.
Immediately it gives relief of the
toothache. The fruit is used to eat in
large quantity as abortifacient.
Zingiber cassumunar (Zingiberceae) Roxb.Adi/RGU/G-
20/ Kekir
Rhizome Tooth ache,
cough, post
delivery pain
relief
The rhizome is boiled with water;
this extract is given to sip by the
person for getting relief of cough.
Medicine is applied in 3–4 times a
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day for 2–3 days. During post
delivery pain; the cooked Rhizome is
given to mother in a low dose.
Raphidopora spp (Araceae) Adi/ RGU/BF-21/ Loma losut
Stem and
leaves Bone fracture The plants to be applied is collected
and processed in the crude form.
Leaves or stems are collected,
cleaned and crushed, making a paste
of the plant or part thereof. The paste
is applied to the specific part of the
bone fracture. At every 24 hours the
paste is reapplied replacing the old
one. The process is repeated for
specific period as suggested by the
“Miri” (the local healer).
Entada scadens (Mimosaceae)
Adi/ RGU/BF-22/ Taboh
leaves
Bone fracture
Phyllostachys assamica (Poaceae) Adi/RGU/BF-23/Tempor
Stem and
Bark Bone fracture
Equisetum arvernse/diffusum
(Equisetaceae) Adi/ RGU/BF-24/ Sisi dungki
Stem Bone fracture
Ricinus communis L
(Euphorbiaceae) Adi/ RGU/BF-25/ Aki-Rokmic
Bark and
leaves Bone fracture
Annanus comosus (Bromeliaceae) Adi/ RGU/RC-26 /Anaros, Tako
bela
Fruit Fertility control in
women. (abortifacient)
Fruit is used to induce abortion.
Dysoxylum alliarum (Maleaceae) Adi/ RGU/RC-27/ Situ paiyu
Bark Fertility control in Pigs and Dogs
(abortifacient)
The bark is dried in sunlight. The
dried bark is grinded and the fine
powdered form is given to the
domestic animals mixing with other
food.
Hemidesmus indicus
(Apocynaceae) Adi/ RGU/RC-28/
Stem Fertility control in
women. (abortifacient)
Stem portion is shade dried to make
powder. The powder id eaten with
water.
Mimosa pudica (Mimosaceae)
Adi/ RGU/RC-29/ Anying ing
Whole
plant Fertility control in
women Fresh juice extracted from the whole
plant taken orally with other food by
the desired woman to avoid
pregnancy.
Scoparia dulcis
(Scrophulariaceae) Adi/ RGU/RC30/Meetagoss.
Arial part Fertility control in
women Fresh juice of aerial part taken orally
by the desired women to prevent
pregnancy.
Calotropis gigantea (Apocynaceae) Adi/ RGU/RC-31 / Abok
Roots Fertility control in
women Calotropis gigantea, Carica papaya
(seeds), Coptis teeta are used in
composite form (paste). This
preparation is given to the desired
women for prevention of pregnancy Carica papaya (Cariaceae) Adi/ RGU/RC-32/ Omir
Mature
seeds Fertility control in
women
Dioscoria allata (Dioscoraceae) Adi/ RGU/RC-33/ Janghli allu.
Tuber Fertility control in
women The tuber is eaten in uncooked form.
It has been stated that eating
frequently by desired women to lose
fertility.
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Figure 3. Percentage of use of TM for various ailments and diseases
Figure 4. Percentage of TM use
DISCUSSION
During the last few years attention of the
world communities has been drawn on
conservation and protection of Biological
diversity of the world. Emphasis was put on the
conservation, protection and sustainable use of
TM in the Convention on Biological Diversity
(World Health Organization, 1999; Note by the
Executive secretary, Secretariat of the
Convention on Biological Diversity. Geneva,
November, 2000). India has an age old tradition
of medicinal practice especially of herbal origin
in Ayurveda. More than 6000 medicinal plant
species has been used in preparation of
medicines by the traditional medicine
practitioners in Indian subcontinent (Seth et al.,
2004). Arunachal Pradesh is one of the north
eastern provinces of India and recognized as the
eastern Himalayan Mega Biodiversity hotspot
(Myers N et al., 2000). The state is situated on
the top of eastern Himalayan Mountain range
having more than eighty percent forest coverage
of the total geographic area of the state. The
biodiversity of the state not fully explored due to
dense forest, very thin population distribution
and lack of infrastructure for surface
communication. Despite all the difficulties,
efforts have been made by some authors to
study and document the ethno medicinal
practice of these indigenous people. Till date
over 500 species of medicinal plant has been
reported from Arunachal Pradesh (Arunachal
Pradesh Information bulletin no.12, 2011).
Traditional medicines among the Chakma,
Singpho, Tangsa, Nishi, Hillmiri, Sulung and
Apatani (Nath et al., 1993; Pal GD, 1984) tribes
of Arunachal Pradesh were reported earlier in
different scientific studies. A few plants used
for traditional medicines among the Adi people
have been reported elsewhere (Gangwar et al.,
1990). These studies showed that plants like
Alstonia scholaris, Calotopris gigantea and
Dioscorea alata which are used by the Adi tribe
are also reported to use by Apatani tribe (Kala
CP, 2005) of Arunachal. On the contrary to the
use of Dioscorea alata as anti-fertility medicine
by the Adi tribe, the Monpas and Apatani use it
for gastritis and indigestion respectively (Nima
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D et al., 2011 and Kala CP., 2005). However,
species Psidium guajava is found to be used by
both the Adi and Monpas for the treatment of
diarrhea (Nima D et al., 2011). The study of the
present authors on traditional medicines is an
added information in wider areas of common
ailments like diarrhea, malaria, eye infection,
wound healing etc as well as joining of bone
fracture and reproduction control prevail among
the Adi people living in remote villages of East
Siang District. The present study revealed a
good number of plants used for fertility control
practice by Adi people of East Siang district. It
has been speculated that practice of herbal
medicines used for fertility control originated in
the population control requirement of domestic
animals. In fact, the Adi people use the anti-
fertility herbal preparation for control of
population growth of their domestic animals. In
this context it is to be mentioned that
maintaining domestic pets (dogs and pigs) is a
cultural entity of the Adi people. Bigger family
size of domestic animals leads to the
requirement of more feed and space. Very often,
owners are unable to provide necessary food or
space to the growing members of their domestic
animals. To control the population growth of
the domesticated animal they regularly practice
such use of plants. These antireproductive
medicines are used during the first trimester of
pregnancy. Once the owner observes mating of
animals; they fed the herbal formulation to the
female to avoid giving birth of new born. These
herbal formulations used during post coital
period may possess abortifacient / antizygotic,
and/ or anti implantation properties. The present
study revealed that these medicines are
successfully utilized by these folk people to
control the reproduction of their domestic pets
and very often to control the unwanted
pregnancies of women. The in-vivo testing of
Dysoxylum alliarium and Piper species showed
pregnancy disruption activity in albino rats
(Das M et al., 2013, Ali N & Ghosh B, 2007,
Mibang Tamo and Chowdhari SK, 2003).
During the survey 15% of total collected
plants have recorded for its use in treatment of
bone fracture. Discussion with the local people
revealed that bone fracture is a common
phenomenon among the villagers as because
the rural folk people have to walk in the steep
hills for livelihood earning. There is no modern
facility for treatment of fractured bone in the
remote areas. This leads to the use of traditional
medicines for recovery from these ailments in
the remote villages. Moreover, malaria,
diarrhea, typhoid are some of the common
health problems faced by these people who use
TM for disease cure and prevention. These
diseases are found among the people who
suffer from unavailability of pure drinking
water and live in unhygienic environment. It
has been found that a single plant is used for
more than one health problem or condition;
such as the Alstonia scholaris (malaria, wound
infection and abortion), Coptis teeta (malaria
and abortion) and Zanthoxylum hamiltoniamun
(toothache and abortion). Medicinal use of
Zanthoxylum spp. for various purpose e.g. use
in malaria (Singh HB & Singh TB., 2005),
relief of labor pain and safe childbirth by
women (Qureshi et al., 2006) as well as use in
toothache (Cheryl Lans, 2007) has been
documented earlier. The Carica papaya has
been reported to use in composite form with
other plant parts for abortion (Valsala S,
Varpagaganapathy P.R. 2002). Important to
note that some of the plant species used by the
Adis of East Siang are reported for its
medicinal use from other states of India as well
as other countries of the world. Mimosa pudica
and Scoparia dulcis which is widely used by
Adi people are also reported to use by the
people of Trinidad and Tobago for treatment of
reproductive disorders of both males and
females and during childbirth (Ignacimuthu et
al., 2006). These folk people of Trinidad and
Tobago reported to use the plant Mimosa
pudica for treatment of women‟s menstrual
cycle disorder (Cheryl Lans, 2007). The
medicinal properties of Solanum nigrum and
Alstonia scholaris is reported to be used by
other tribes of India. Solanum nigrum is used
for wound healing, stomachache and also as an
abortive (Ignacimuthu et al., 2006, Sen et al.,
2012) while, Alstonia scholaris is used for
increase lactation in women (Nain Jaspreet et
al., 2011). The Adi people developed the
practice of using composite medicines. More
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 298–310
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than one plant species combined together; e.g.
the combination Calotropis gigantea, Coptis
teeta, Dioscorea allata are used as abortifacient
drug. The justification for use of composite
medicines is not known by the TMPs. As
mentioned earlier, practice of these medicines
by the local TMPs is associated with
superstitions and worships of their deity. The
epistemology of the traditional practice of
preparation and use of these TM needs to be
reviewed by the epistemology of scientific
knowledge.
Some of the TMPs, the authors interacted
were well educated and aware of the need of
conservation of the traditional knowledge and
the conservation of plant species. Many of the
practitioners were maintaining their own
medicinal plant garden for conservation and
regular use for medicine formulation. Proper
laboratory screening of these traditional herbal
preparations may come across through
development of new lead, useful for cure of
various ailments/ conditions as well as for
fertility control. This approach shall help in the
development of pharmaceutical industry,
production of cheaper health friendly drugs for
the rural people especially of the developing
countries and rural economic empowerment.
ACKNOWLEDGEMENT
The authors acknowledge the funding
received under the scheme “Women Scientist
Scholarship Scheme for Societal Programme
(WOS-B), Department of Science and
Technology, Government of India” for
carrying out this research. The authors are also
grateful to the University Grants commission,
New Delhi for providing necessary support to
undertake this research under the Center with
Potential for Excellence in Biodiversity, Rajiv
Gandhi University. We extend our thanks to
Osiri Ratan, Nanku Gao,Y.J Lego and other
TMPs for sharing their knowledge on
traditional medicine and help in various ways.
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Source of Support: Women Scientist
Scholarship Scheme for Societal
Programme (WOS-B), Department of
Science and Technology, Government of
India
Conflict of Interest: None Declared
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 311–316
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ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
PRELIMINARY PHYTOCHEMICAL SCREENING OF SECONDARY
METABOLITES OF ADHATODA VASICA NEES. FLOWERS
Chouhan Surksha¹, Bhatt Shashank²*, Dhyani Suresh3
1Department of Biotechnology, Rajiv Gandhi Govt. P.G. College, Mandsaur, Madhya Pradesh, 458001,
India. 2,3
Department of Biotechnology, NIMS University, Jaipur, Rajasthan, 303121, India. *Corresponding author: Email: [email protected]; Mobile: +919826840428
Received: 02/04/2013; Revised: 25/04/2013; Accepted: 30/04/2013
ABSTRACT
Adhatoda vasica Nees. belongs to the family Acanthaceae. The work was done on Adhatoda
vasica Nees. flowers and different solvents were used for extraction of the phyto-constituents.
Solvents like petroleum ether, chloroform, methanol, 95% ethanol and distilled water were selected
for extraction of the flowers according to their polarity. Alkaloids, flavonoids, glycosides, saponin,
phenol, lignin, sterols and tannins were found in phyto-chemical screening. These metabolites have
different types of medicinal activities as antibacterial, antiviral, anti-diabetic etc. which could be
efficiently used in treating various ailments.
KEY WORDS: Adhatoda vasica Nees., Alkaloids, Glycosides, Sterols, Lignins.
Research article
Cite this article:
Chouhan Surksha, Bhatt Shashank, Dhyani Suresh (2013), PRELIMINARY PHYTOCHEMICAL
SCREENING OF SECONDARY METABOLITES OF ADHATODA VASICA NEES. FLOWERS,
Global J Res. Med. Plants & Indigen. Med., Volume 2(5): 311–316
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 311–316
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
INTRODUCTION
Plants can be categorized according to their
properties as medicinal and wild. Near about
80% plant species’ compounds have medicinal
activities and used as medicine (WHO, 1993).
In India, 45,000 plant species are officially
recorded and 7500 medicinal plant species
grows in 16 agro-climatic zones under 63.7
million hectares of forest coverage (H. Tag,
2007). Most of the plants have been involved
into medicinal category.
Plants are the natural sources for the
development of medicines (Kumar, 2004).
Metabolites found in plants show an important
role in the treatment of diseases. Metabolites
are falls into two categories one of which is
primary and another is secondary metabolites.
Primary metabolites play important roles in
metabolic pathways while secondary
metabolites support it. These secondary
metabolites are alkaloids, flavonoids,
glycosides, saponins, lignin etc.
Adhatoda vasica Nees. commonly known
as Malabar nut belongs to the family
Acanthaceae. It is an evergreen shrub and
grows in waste places. It is distributed through
out in India and utilized in rural areas for
different ailments (K. Jayapaul et al., 2005).
The stem, leaf, flower, fruit and seeds have
drug properties (B.L. Manjunath, 1948, U.P.
Claeson et al., 2000). It has antispasmodic,
fever reducing, anti-inflammatory, anti-
bleeding, bronchodilator, anti-diabetic, anti-
helminthic, disinfectant, anti-jaundice,
antiseptic, expectorant and many other
medicinal uses (P.K. Patel et al., 1984, A.
Chakraborty et al., 2001, R. L. Wakhloo et al.,
1980).
MATERIAL AND METHODS
Collection of Plant Material
Adhatoda vasica Nees. is found all over the
world. The plant flowers were collected from
Mandsaur district, Madhya Pradesh. Mandsaur
District forms the northern projection of
Madhya Pradesh (Fig.-1). It lies between the
parallels of latitude 23° 45' 50" North and 25°
2' 55" North, and between the meridians of
longitude 74° 42' 30" East and 75° 50' 20" East.
Figure 1: Map of Madhya Pradesh.
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Figure 2: Adhatoda vasica Nees.
Preliminary Screening of Secondary
Metabolites
The flowers of Adhatoda vasica Nees. (Fig.
2) were collected from Mandsaur Madhya
Pradesh and plant identified to Dr. S.N. Mishra,
Principal Scientist, All India Coordinated
Research Project on Medicinal and Aromatic
Plants, College of Horticulture, Mandsaur,
affiliated to R.V.S.K.V. Vishwavidyalaya,
Gwalior.
The flowers were shade dried, powdered
using mixer grinder, and subjected to cold
percolation process for 48 hours with
petroleum ether, chloroform, methanol and
distilled water. After this process, the extracts
were filtered and used for preliminary
phytochemical screening such as alkaloids
(Iodine, Wagner, and Dragendorff’s test),
flavonoids (Pew’s, Shinoda and NaOH tests),
glycosides (Keller-Killani, Conc. H2SO4, and
Molisch tests), lignin (Labat and Lignin tests),
phenols (Ellagic acid and Phenol tests),
saponins (Foam and Haemolysis test), sterols
(Libermann- Burchard, and Salkowski tests),
tannins (Gelatin and Lead acetate tests) were
carried out (Shashank Bhatt et al.,2011).
Preliminary Screening of Phyto-chemical
Test
Phytochemical Screening
The filtrate obtained was subjected to
preliminary phyto-chemical screening.
For identification of alkaloids Iodine test
(Khandelwal K.R., 2008), Wagner’s test
(Kokate C.K. et al., 2001) and Dragendorff’s
test (Kokate C.K. et al., 2001) were done.
For identification of flavonoids Pew’s test
(Peach K., Tracey MV. 1956), Shinoda test
(Kokate C.K. et al., 2001) and NaOH test
(Khandelwal K.R., 2008) were performed.
For identification of glycosides Keller-Killani
test (Kokate C.K. et al., 2001), Glycoside test
(Treare GE, Evans WC. 1985), Conc. H2SO4
test (Khandewal K.R., 2008) and Molish test
(Kokate C. K. et. al., 2001) were done.
For identification of Phenol Ellagic acid test
(Gibbs R.D., 1974) and Phenol test (Gibbs
R.D., 1974) were done.
For identification of lignin lignin test (Gibbs
R.D., 1974) and labat test (Gibbs R.D., 1974)
were performed.
For identification of saponins foam test
(Kokate C. K. et al., 2001), and Haemolysis
test (Kokate C.K., 1994) were performed.
For Identification of sterols Liebermann-
Burchard test (Kokate C. K. et al., 2001) and
Salkowski’s test (Kokate C. K. et al., 2001)
were done.
For identification of tannins gelatin test (Trease
GE, Evans WC. 1985) and lead acetate test
(Trease GE, Evans WC. 1985) were done.
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TABLE-1: Phytochemical Study of Adhatoda vasica Nees. Flowers
Test Petroleum
Ether
Chloroform Methanol 95% Ethanol Distilled Water
Alkaloids
Iodine Test − − − − −
Wagners Test − − + + −
Dragendorff’s
Test
− − + + −
Flavonoids
Pews Test − − + + −
Shinoda Test − − + + +
NaOH Test − − + + +
Glycosides
Keller- Killani
Test
− − + + +
Glycosides Test − − + + +
Conc. H2SO4 − − + + +
Molishs Test − − + + +
Phenol
Ellagic Test − − + + −
Phenol Test − − + + −
Lignin
Lignin Test − − + + −
Labat Test − − + + −
Saponins
Foam Test − − + + −
Haemolysis Test − − + + −
Sterols
Libermann-
Burchard Test
− + + + +
Salkowski Test − + + + +
Tannins
Gelatin Test − − + + +
Lead Acetate Test − − + + +
RESULT & DISCUSSION
The flowers were powdered and subjected
to cold percolation with petroleum ether,
chloroform, methanol, 95% ethanol and
distilled water for 48 h. and found alkaloids,
flavonoids, glycosides, phenol, lignins,
saponins, sterols and tannins. Adhatoda
vasica’s flowers have most of the secondary
metabolites that mention in table- 1. Each
metabolite has specific activities. Therefore, it
may be effective.
Flavonoids, tannins, saponins, steroids and
phenols have been used in the treatment of
different types of diseases. These moieties are
isolated and extracted from medicinal plants’
parts and used for development of new drug.
If flowers are selected in research most of the
secondary metabolites can be found easily and
isolate it for development of new drugs. We
know that steroids are also involved into
hormonal category which is responsible for
changes in male and female characteristics.
Tannins have general antimicrobial and
antioxidant activities (Rievere et al., 2009).
Current reports show that tannins may have
potential value such as cytotoxic and anti-
neoplastic agents (Aguinaldo et al., 2005).
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 311–316
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CONCLUSION
I have concluded that Adhatoda vasica
Nees. flowers have most of the secondary
metabolites that have different activities by
which diseases treated. The specifications of
Siddha and Ayurveda show the limit of
medicinal plant categories. The Ayurveda and
Siddha have shown the uses of this plant parts
but not involve the flowers. Each part of plant
has not all secondary metabolic compounds
that have concluded but only flower have it.
Therefore, according to my results, I have
concluded that its flowers may be most useful
part to another part of plant.
ACKNOWLEDGEMENT
I am very thankful to Dr. S.N. Mishra for
assist in identification of plant and kind
assistance of my dear father Shri Krishna
Kumar Bhatt, Smt. Subhadra Bhatt and brother
Mr. Mayank Bhatt.
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Source of Support: Nil Conflict of Interest: None Declared
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 317–336
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ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
HERBAL MEDICINES FOR DEPRESSION AND ANXIETY: A
COMPREHENSIVE STATE OF THE ART REVIEW
Patel Shanti1, De Sousa Avinash
2*
1Medical Intern – Lokmanya Tilak Municipal Medical College and General Hospital, Mumbai, Maharashtra,
India 2Consultant Psychiatrist and Founder Trustee – De Sousa Foundation, Mumbai- 400054, Maharashtra, India
*Corresponding Author: E-mail – [email protected]; TEL – 91-22-26460002
Received: 01/03/2013; Revised: 26/04/2013; Accepted: 30/04/2013
ABSTRACT
This review looks at all the herbal medicines and formulas in treating depression and anxiety
disorders. Pubmed and the Cochrane Library were searched for pharmacological and clinical
evidence of herbal medicines with antidepressant and anti-anxiety action. Good evidence exists for
the use of kava and St John‟s wort in the treatment of anxiety and depression respectively, while
there is insufficient clinical evidence for the use of many other herbal medicines in psychiatric
disorders. Newer herbal preparations that potentially have significant use in depression and anxiety
and urgently require more research are Rhodiola rosea (roseroot), Crocus sativus (saffron),
Passiflora incarnata (passion flower) and Piper methysticum (kava). They need further evidence
base via clinical studies. Depression and anxiety are commonly researched but the efficacy of herbal
medicines in these disorders requires attention. The review addresses all the current issues in herbal
therapy, safety issues and future areas of application in the field.
KEY WORDS: Herbal medications, depression, anxiety, kava, St John‟s wort, passion flower.
Review article
Cite this article:
Patel Shanti, De Sousa Avinash (2013), HERBAL MEDICINES FOR DEPRESSION AND ANXIETY: A
COMPREHENSIVE STATE OF THE ART REVIEW, Global J Res. Med. Plants & Indigen. Med.,
Volume 2(5): 317–336
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 317–336
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
INTRODUCTION
Mood disorders, anxiety and sleep disorders
are largely prevalent and highly comorbid
psychiatric conditions (Kessler et al., 2005). It
is estimated that by 2020 depression will result
in 2nd
greatest increase in morbidity after
cardiovascular diseases, presenting a significant
socioeconomic burden (WHO, 2006). Since the
past decade, many herbal medicines have been
used in people with mood and anxiety disorders
(Schulz et al., 2001). Due to the increasing
popularity of herbal medications majority of
the patients are consulting herbalists,
naturopaths, and other healers, in addition to
physicians. A data from a nationally
representative sample of 2055 people
interviewed during 1977–1988 revealed that
57% of those suffering anxiety attacks, and
54% of those with severe depression reported
using herbal medicine during the previous 12
months to treat their disorder (Kessler et al.,
2001). Similarly interviews of 82 psychiatric
North American inpatients revealed that 44%
had used herbal medicine (mainly for
psychiatric purposes) during the previous 12
months (Elkins et al., 2005).
There is however, a limited data regarding
the benefits and liability of herbal remedies and
other natural remedies. There have been few
reports of serious adverse effects from these
medications and by and large these medications
have been considered safe and effective
(Schulz et al., 2001; Mischoulon, 2004). This
article reviews the literature on various herbal
medications in the treatment of depression and
anxiety.
MECHANISM OF ACTION OF HERBAL
MEDICATIONS
The primary mechanism of action involves
modulation of neuronal communication, via
specific plant metabolites binding to
neurotransmitter/neuromodulator receptors
(Spinella, 2011) and via alteration of
neurotransmitter synthesis and general function
(Sarris, 2007). Other mechanisms involve
stimulating or sedating CNS activity, and
regulating or supporting the healthy function of
endocrine system (Kumar, 2006; Sarris, 2007;
Spinella, 2011). The psycho-pharmacological
effects of herbal medicines and their clinical
validation can be explored by the use of “omic”
genetic technologies (Ulrich-Merezenich et al.,
2007).
HERBAL MEDICATIONS USED IN THE
MANAGEMENT OF DEPRESSION
Hypericum perforatum L. (St.John’s wort)
For centuries, hypericum an extract of the
flower of St. John‟s Wort (SJW) (Hypericum
perforatum L.) is used for the treatment of
depression (Schulz et al., 2001). Its use in the
United States has been dramatically increased
in the past decade. Polycylic phenols, hypericin
and pseudohypericin are the active compounds
in extract of St. John‟s Wort. Other compounds
include flavonoids (hyperoside, quercetin,
isoquercitrin, rutin), kaempferol, luteolin,
biapigenin and hyperforin (Muller-Kuhrt and
Boesel, 1993; Staffeldt et al., 1993; Wagner et
al., 1993). Out of all the active compounds
hypericin is the main active compound.
Hypericin decreases serotinin receptor
density (Muller-Kuhrt and Boesel, 1993). It
also inhibits monocyte production of
interleukin 6 and 1β resulting in a decrease in
corticotropin releasing hormone and thus
dampening cortisol production (Thiele et al.,
1993). It decreases expression of β
adrenoreceptors and increases density of
serotonin by nonselective inhibition of neuronal
reuptake of serotonin, dopamine,
norepinephrine, GABA and l-glutamate,
decreased degradation of neurochemicals, and a
sensitization of and increased binding to
various receptors (e.g. GABA, glutamate and
adenosine) (Butterweck, 2003; Mennini and
Gobbi, 2004; Zanoliu, 2004; Muller et al.,
1993; Teufel-Mayer et al., 1997). SJW
modulates salivary and serum cortisol levels,
and has a slight effect on growth hormone
(Franklin et al., 2006). Hyperforin, hypericin
and various flavonoids appear to be responsible
for the neurochemical modulation (Butterweck,
2003; Laakmann et al., 1998; Zanoliu, 2004).
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In a large number of clinical European
clinical trials hypericum has been compared
with low dose imipramine and maprotiline
(75 mg/day) (Varbach et al., 1994; Harrer et
al., 1993). Despite these low doses of active
controls, the response rates in these trials
seemed comparable to those in studies that use
higher doses of tricyclic antidepressant agents
(TCAs) (eg. Imipramine ≥ 150 mg/day). The
response rates for hypericum ranged from
35.3% to 81.8%, and the response to TCAs
ranged from 41.2% to 77.8%. In a meta-
analysis (Nirenberg et al., 2002), hypericum,
300 mg three times a day was judged to be
effective in 79 of 120 subjects (65.8%),
whereas placebo was considered effective in
only 36 of 125 subjects (28.8%). The placebo
response rate seemed comparable to that
observed in many outpatient studies of anti
depressants conducted in United States. A
recent meta-analysis conducted by Rahimi et
al., (2009) yielded a significant relative risk
(RR) for response in favour of the active of
1.22 (95%Cl :1.03, 1.45) and weighed a mean
difference between treatments of 1.33 points
(95%Cl: 1.15, 1.51) on the Hamilton
Depressing Rating Scale (HAM-D). Where as
comparison with SSRIs yielded a non
significant difference between treatments of
0.32 (95% Cl: - 1.28, 0.64) for mean reduction
in HAM-D score from baseline.
A meta-analysis (Linde et al., 1996)
examined 15 trials comparing Hypericum with
placebo and eight trials comparing Hypericum
with TCAs in 1757 patients who had mild to
moderate depression.In six trials that used
single preparation of Hypericum, (containing
only St.John‟s Wort), hypericum yielded
greater response rates than placebo (55.1% for
Hypericum versus 22.3% for placebo) and
comparable response rates to tricyclic
antidepressants (69.3% for Hypericum versus
58.5 & for tri-cyclic antidepressants). In two
trials that used combination preparations of
Hypericum (containing St.John‟s wort and
other herbal medications such as Kava,
Hypericum was found to be more effective than
TCAs (67.7% versus 50%).
In a 6 week trial with 375 patients,
Lecrubier and colleagues (Lecrubier et al.,
2002) found that St. John‟s Wort, 900 mg/day,
was significantly more effective than
placebo,especially who had higher base line
HAM-D scores. Shelton and colleagues (2001)
found that St.John‟s Wort (900–1200 mg/day)
was no more effective than placebo in the full
intent to treat analysis, although among
completers the remission rates were
significantly higher with St. John‟s Wort than
the placebo. A 2004 meta-analysis of SJW
(dosage 300–1200 mg/day) in the treatment of
mild to moderate depression (Roder et al.,
2004) reviewed 30 trials and concluded a
significant advantage of SJW over placebo
(n=2129, relative risk , RR = 0.66,95% Cl 3.0
to 6.6, mean response 53.2 SJW vs 51.3 %
synthetic antidepressants).A meta-analysis of
16 trials ,inspection of individual studies
showed that SJW was found to demonstrate
greater efficacy than synthetic
antidepressants.Six RCTs tested SJW against
placebo and fluoxetine in treating MDD, as
commonly assessed via the Hamilton rating
scale for depression (HAM-D) and clinical
global impression (CGI). Four studies
demonstrated that SJW had similar (Behnke et
al., 2002; Bjerkenstedt et al., 2005) or superior
(Fava et al., 2005; Schradar, 2000) effects to
fluoxetine. An analysis of the sub-sample of a
12-week 3-arm study discovered that SJW
(160- 900mg/day) ameliorated depression –
based vegetative presentations, while fluoxetine
(20mg/day) was statistically equivalent to
placebo (Murck et al., 2005).
In comparison to paroxetine, SJW was
statistically more effective in treating
moderate-severe depression (Szegedi et al.,
2005). Other comparative trials demonstrated
SJW‟s statistical ewuivalence to imipramine
(Woelk, 2000), citalopram (Gaspere et al.,
2006), Maprotiline (Harrer et al., 1994) and
amitriptyline (Wheatley, 1997) in treating
major depressive disorder. The Hypericum
Depression Study, the medicine is currently
used for the treatment of mild to moderate
depression (Clement et al., 2006; Lawvere and
Mahoney, 2005; Linde et al., 2005).
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A comparative analysis between paroxetine
and Hypericum extract WS 5570 revealed that
paroxetine had 10 to 38 times higher adverse
event rate.An increasing number of averse drug
reaction have been noted between St.Johns
wort and other medications.Majority of the
interactions are due to the liver enzyme CYP-
450-3A4 which results in the decreased activity
of several drugs, including warfarin,
phenprocoumon, digoxin, indinavir,and
irinotecan (Baede-van Dijk et al., 2000; Miller
et al., 1998; Moore et al., 2000; Miller et al.,
2000; Piscitelli et al., 2000). The interactions
are mainly due to high dose hyperforin extracts
(Izzo, 2004). Hyperforin increases the
expression of pregnane-X-receptor,which
increases P-glycoprotein expression (Dresser et
al., 2003; Izzo, 2004; Moore et al., 2000). Low
hyperforin preparations may not affect this
response and hence may be safer (Izzo 2004,
Muller et al., 2006). A systematic review of 19
studies showed that high dose hyperforin
extracts (>10mg/day) had outcomes consistent
with CYP3A induction while studies using low
dose hyperforin extracts (<4mg/day)
demonstrated no significant effects on CYP3A
(Whiten et al.,2004). Because of the
monoaminase inhibiting ativity of St.John‟s
Wort, its combination with SSRI‟s may result
in Serotonin syndrome, hence it should not be
combined with SSRI‟s (Hu et al., 2005). As
monotherapy adverse effects are mild (Schulz,
2005). Adverse events include dry mouth,
dizziness,constipation, other gastrointestinal
symptoms and allergic reactions.(Schulz,2001;
Schulz, 2005).
Phototoxicity has been found in animals
with hypericum but rarely in humans.
Hypericum at a dose of 1800mg caused minor
increase in sensitivity to uv light in humans but
no phototoxicity. It is recommended that
patients taking high dose of hypericum should
be isolated from UV radiation for 7 days
(Seigerse et al., 1993). At least 17 cases of
psychosis have been resulted from St, John‟s
Wort ,of which 12 comprised mania or
hypomania. Researchers compared St.John‟s
Wort, 900 to 1800mg/d with sertraline 50 to
100mg/d, in 12 community based primary care
offices. It was found that St John‟s Wort
resulted in significantly fewer adverse events
(Van Gurp et al., 2002). In a 2006 review of 16
post marketing surveillance studies (n=34834)
(Schulz, 2006), SJW was deemed to be 10 fold
safer than synthetic antidepressants (adverse
effects 0.1% to 2.4%). Overall SJW has
demonstrated equal efficacy to pharmaceutical
antidepressants with a more favourable side
effect profile and fewer dropouts than its
synthetic counterparts. SJW has been
recommended as a first line treatment in milder
forms of depression (Roder et al., 2004).
Crocus sativus L. (Saffron)
It increases the re-uptake inhibition of
monoamines (dopamine, norepinephrine and
serotonin). It is also a NMDA receptor
antagonist and a GABA-α receptor agonist
(Hosseinzadeh and Noraei, 2009; Lechtenberg
et al., 2008; Schmidt et al., 2007). There have
been two trials (Akhondzadeh et al., 2004;
Noorbala et al., 2005) comparing saffron with
imipramine and fluoxetine, it was found that
saffron demonstrated improvement of
depression. A similar response was
demonstrated in a study in which 30 mg saffron
was effective over placebo (Akhondzadeh et
al., 2004; Moshiri et al., 2006). Clinical trials
have detailed anxiety, tachycardia, nausea,
dyspepsia and changes in appetite as possible
side effects (Moshiri et al., 2006).
Lavendula spp. (Lavender)
It causes GABA modulation. In animal
studies it is effective in anxiety symptoms
(Atsumi and Tonosaki, 2007; Bradley et al.,
2007; Perry and Parry, 2006; Shaw et al., 2007;
Toda and Morimoto, 2008). In a 4 week RCT
comparing comparing lavender tincture (1:5
50% alcohol,60 drops) against imipramine in
patients (n=45) with a HAM-D rating of atleast
18 it was found that although lavender was less
effective than the synthetic counterpart ,the
combination of both was more effective than
Imipramine alone,indicating a synergistic effect
(Akhondzadeh et al., 2003).
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Rhodiola rosea L. (Rose root)
It causes inhibition of cortisol, stress
induced protein kinases, nitric oxides and
Monoamine oxidase A. In animal models it has
shown to cause normalization of 5-HT and anti
stress effects (Chen et al., 2009; Panossian et
al., 2007; Panossian et al., 2008; Mattioli et al.,
2009; Perfumi and Matticki, 2007; Van
Dierman et al., 2009). Authors (Schevtsov et
al., 2003) assessed the influence of roseroot on
various mental and biological parameters of
161 adults, it was found to have an ant fatigue
effect. This property along with the monoamine
modulation can be used in the treatment of
monopolar depression (Stancheva and
Mosharrof, 1987). A three-arm study using R.
rosea 5HR-5 standardised extract (340 mg and
680 mg/day) against placebo in the treatment of
mild-moderate depressive disorder revealed a
significant dose dependent improvement
occurred in the active groups compared with
placebo (Darbinyan et al., 2007).
S-Adenosyl Methionine (SAMe)
It is a methyl donor in the brain and is
involved in the pathways for synthesis of
hormones, neurotransmitters, nucleic acids,
proteins and phospholipids. Its potential role in
mood regulation was determines by its activity
as an intermediate in the synthesis of
norepinephrine, dopamine, and serotonin. It is
used in the treatment of major depression as
well as in other medical conditions (Spillmann
et al., 1996). Depression has been associated
with Folate and vitamin B12 deficiency and
about 10% to 30% of depressed patients have a
low folate and these patients respond less to
antidepressants (Alpert et al., 2000). Vitamin
B12 is converted to methylcobalamin which is
involved in the synthesis of various
neurotransmitters. Hence its deficiency may
result in earlier age of onset of depression
(Fava et al., 1997). SAMe is synthesized from
the amino acid l-methionine through the one
carbon cycle, a metabolic pathway involving
the vitamins folate and B12 (Spillmann et al.,
1996). Low SAMe levels have been found in
cerebrospinal fluid of depressed individuals
(Bottiglieri et al., 1990) and higher plasma
SAMe levels have been associated with
improvement in depressive symptoms (Bell et
al., 1994). It has been found that folate
augmentation in partial responders has
achieved good results (Coppen et al., 2000;
Alpert et al., 2002). In 8 placebo controlled
studies SAMe demonstrated superiority to
placebo in 6 studies and equivalency to placebo
in the other 2 studies (Spillmann et al., 1996;
Alpert et al., 2000; Coppen et al., 2000). In 6 of
the 8 comparison studies with TCAs, SAMe
was equivalent in efficavy to TCAs and was
more effect than imipramine in one study and
SAMe may have a relatively faster onset of
action than conventional depressants
(Spillmann et al., 1996; Alpert et al., 2000;
Coppen et al., 2000). In one study, some
patients improved within a few days,and most
did so within 2 weeks.(Fava et al., 1995). Two
studies have shown that combination of SAMe
and a low dose TCA resulted in earlier onset of
onset than a TCA alone (Alvarez et al., 1987;
Berlanga et al., 1992).
Researchers have examined the efficacy of
SAMe as an adjunctive treatment for partial
and nonresponders to SSRIs (Alpert et al.,
2004). Thirty subjects who had residual
depression despite SSRI or venlafaxine
treatment received a 6-week course of 800–
1600mg. Response and remission rates with
SAMe augmentation were 50% and 43%
respectively, and the treatment was well
tolerated. Besides depression, SAMe is
effective for dementia-related cognitive defects,
depression in patients who have parkinson‟s
disease or other medical illness, psychological
distress during the puerperium and opoid and
alcohol detoxification (Mischoulon et al.,
2002). SAMe is well tolerated and relatively
free of side effects. Side effects include mild
insomnia, lack of appetite, constipation, nausea,
dry mouth, sweating, dizziness and nervousness
(Spillmann et al., 1996). Cases of increased
anxiety, mania or hypomania in bipolar
depression have been reported (Spillmann et
al., 1996; Carney et al., 1987; Carney et al.,
1983) and therefore it should be used carfully
in patients with bipolar disorders.
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Omega-3 fatty acids
The intake of more and more processed
foods rich in omega-6 containing vegetable oils
has decreased the intake of omega- 3 fatty acids
in the Western diet. This has resulted in higher
physiologic ratio of omega-6:omega-3 fatty
acids in Western countries (Adams 1996;
Hibbeln, 1995; Cross-National Collaborative
Group, 1992; Hibbeln, 1998; Hibbeln, 1999). It
has been postulated that the modern western
diet and the additional stresses of twenty-first
century create a proinflammatory state in
humans that may contribute to cardiovascular
and also may play a role in the development of
mood disorders (Stoll and Lacke, 2002). So
administration of omega-3 supplements may
potentially reverse this proinflammatory state
by correcting the omega-6:omega-3 ratio. It has
an effect on membrane -bound receptors and
enzymes involves in the regulation of
neurotransmitter signaling,as well as regulation
of calcium ion influx through calcium channels
(Stoll and Lacke, 2002). Omega-3 fatty acids
cause decrease corticosteroid release and
dampen mood-altering effects associated with
cortisol by inhibiting secretion of inflammatory
cytokines. Eicosapentanoic acid resembles
amitriptyline in antidepressant action, it inhibits
the synthesis of prostaglandin E2, thus
dampening the synthesis of p-glycoprotein
(Murck et al., 2004). Peet and Horrobin (2002)
conducted a randomized, placebo-controlled,
dose finding study of ethyl-eicosapentaenoate
(EPA) as adjunctive therapy for 70 adults who
had persistent depression despite treatment with
a standard antidepressant. Subjects who
received 1 g/d EPA for 12 weeks showed
significantly higher response rates (53%) than
subjects receiving placebo (29%), with notable
improvement of depressed mood, anxiety, sleep
disturbance, libido, and suicidality. The 2 g/d
group showed little evidence for a drug-placebo
difference, and the 4 g/d group showed a
nonsignificant trend toward improvement.
These results suggest that there may be an
optimal dose of omega-3 that humans require
for maximum benefit, and it is possible that an
overcorrection of the omega-6:omega-3 ratio
with higher omega-3 doses may limit the
antidepressant effect of EPA.
Researchers (Su et al., 2002) conducted an
8-week, double-blind, placebo-controlled trial
comparing adjunctive omega-3 (6.6 g/d)
against placebo in 28 depressed patients.
Patients in the omega-3 group had a significant
decrease in HAM-D scores compared with
placebo. Nemets and colleagues found a
statistically significant benefit of adjunctive
EPA in 20 subjects who had major depressive
disorder and who were on antidepressant
therapy, 1 g/d, and a clinically important
difference in the mean reduction of the 24-item
HAM-D scale by the study endpoint at week 4
compared with placebo (12.4 versus 1.6). A
single placebo- controlled study with 36
subjects showed lack of efficacy of DHA, 2
g/d, for depression (Marangell et al., 2003).
Researchers (Osher et al., 2005) treated 12
bipolar I depressed subjects with open
adjunctive EPA, 1.5 to 2 g/d, for up to 6
months. Ten patients completed at least 1
month of follow-up, and eight achieved a 50%
or greater reduction in HAM-D scores. No
cycling occurred with any patients. Further
investigation is needed to determine whether
bipolar disorder actually requires higher doses
of omega-3 fatty acids than unipolar illness and
to unravel the respective contributions of EPA
and DHA. Omega-3 fatty acids are relatively
very safe. Side effects include gastrointestinal
upset and fishy aftertaste tends to occur with
higher doses (> 5 g/d) with less pure
prepararions. At doses of 1 g/d with highly
purified omega-3 preparations, these adverse
effects are less common. There is a
documented risk of bleeding but it is minimal
at doses less than 3 g/d. Hence individuals
taking warfarin should be cautious and should
use omega-3 fatty acids under a physician‟s
supervision. Also, there are few documented
cases of cycling in bipolar patients (Freeman et
al., 2006).
Hence it is recommended hat low doses of
omega-3 fatty acids may be effective and well
tolerated monotherpy or adjunctive therapy for
depressed adults. Freeman and colleagues
(2006) recommends that depressed individuals
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may safely use approximately 1 g/d of an EPA-
DHA mixture but should not substitute omega-
3s for conventional antidepressants at this time.
For, individuals who take more than 3 g/d of
omega-3 should do so under a physician‟s
supervision (Freeman et al., 2006). They can
also be used to treat specific populations (eg.
Pregnant or lactating women) for whom
antidepressants may be used with a caution
(Chiu et al., 2003), for elderly people and for
those with cardiovascular diseases.
Echium amoenum Fisch. & C.A.Mey (Borage)
Its antidepressant action is currently
unknown and anxiolytic activity is shown in
animal studies (Rabbani et al., 2004). In a RCT
single dose of Echium amoenum (375mg/day
for 6 weeks) was compared against placebo in
35 patients with depression and anxiety,
assessed via HAM-D and Hamilton anxiety
scale (HAM-A). It was found that the herbal
medication was superior than placebo in
reducing depressive symptoms with a effect
size d of 0.92 but this result was not maintained
at week 6 (Sayyah et al., 2006). It also had no
anxiolytic activity.
Dan Zhi Xiao Yao
It is a chinese preparation which contains
Dan Pi (Cortex Moutan), Zhi Zi (Fructus
Gardneiae), Chai Hu (Radix Bupleuri), Dang
Gui (Radix Angelicae Sinensis), Bai Shao
(Radix Alba Paeoniae), Bai Zhu (Rhizoma
Atractylodis Macrocephalae), Fu Ling
(Poria) Gan Cao (Radix Glycyrrhizae).It is
modified from Xiao yao san (Rambling
powder) herbal preparation which is used in the
treatment of depression by moving stasis
(Bensky and Gamble, 1991) and in addition it
has Dan zhi (Cortex Moutan). In a RCT, 63
patients with depression assessed via the HAM-
D, self rating depression scale (SDS), self
rating anxiety scale (SAS) and the scale for
TCM syndrome and symptom differentiation,
the formulation was compared with
maprotiline. It was found that maprotiline was
effective in 84% patients in reduction of
depression whereas Dan zhi xizo yao was
effective in 87% (Lou et al., 2006).
Banxia Houpu
It consists of Pinella ternata, Poria cocos,
Magnolia officinalis, Perilla frutescens and
Zingiber officinale. There is no human clinical
data to determine the efficacy of Banxia houpu
but large number of animal studies have
demonstrated its anti-depressant activity
comparable to fluoxetine in trail suspension
and forced swimming tests (Li et al., 2003; Luo
et al., 2000). It was found that an increase in
serotonin (5-HT) and 5-hydroxyindoleacetic
acid (5-HIAA) levels was found to occur in
mouse hippocampus and striatum. Researchers
(Wang et al., 2005) found that Banxia houpe
decoction decreased the level of triglycerides in
serum enhanced the activity of the natural killer
cellsin the spleen, decreased the activity of
superoxide dismutase in red blood cells and the
activity of the nitric oxide synthase in the
serum and the tissue, and reduced the content
of malondialdehyde in tissue via the effect on
lipid peroxidation.
HERBAL MEDICINES USED IN THE
MANAGEMENT OF ANXIETY
Piper methysticum L.f (Kava)
It causes GABA channel modulation (lipid
membrane structure and sodium channel
function) and weak GABA binding which
causes increased synergistic effect of [3H]
muscimol binding of GABA-α-receptors.It also
causes β-adrenergic downregulation and MAO-
B inhibition.It inhibits reuptake of
norepinephrine in prefrontal cortex (Bonon and
Haberlein, 1998; Dacies et al., 1992; Jussofie et
al., 1994; Magura et al., 1997; Uedelhack et
al., 1998). A 2003 cochrane review of
randomosid ,double blind ,controlled trials of
rigorous methodology using Kava mono
preparations (60-280 mg of kavalactones),
Pittler and Ernst found that Kava had a
stastically significant anxiolytic activity on
Hamilton Anxiety Scale (HAMA) compared
with placebo (95% Cl;0.1,7.7) but one trial
demonstrated that kava was effective in short
term treatment of anxiety. A meta analysis
(Sarris et al., 2010b) revealed a similar
conclusion A meta-analysis of 7 homogenous
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trials using HAM-A demonstrated that kava
reduced anxiety significantly than placebo
(weighted mean difference 3.9 over placebo on
the HAM –A;95% Cl:0.1 to 7.7 p=0.05; n-
380). A 4 week study by Connor and Davidson
(2002) found no significant difference between
a standardised Kava extract and placebo.
A meta-analysis beased on six placebo
controlled randomized trials using Kava extract
WS 1490 in anxiety demonstrated that kava
significantly reduced anxiety, with a mean
improvement of 5.94 better than placbo (Witte
et al., 2005). A 3-month randomized
prospective open study investigating kava in
peri menopausal women revealed that the
reduction in anxiety with kava was
significantly greater than in controls (on
calcium supplementation) as assessed via the
State trait anxiety index (STATI). It was also
observed that depression depression declined at
3 months (-5.03+/-1.4) as assessed via the
Zung‟s depression scale (Cagnacci et al.,
2005). A randomized controlled double blind,
multicenter clinical trial compared kava with
synthetic agents like busiprone or opramol
(Boerner et al., 2003). The outcomes were
measured using HAM-A, Boerner anxiety
scale, SAS, CGI, a self rating scale for well
being, a sleep questionnaire, a quality of life
questionnire (QOL) and global judgement by
investigator and patients.It was found there
there was no significant difference between
Kava and Busiprone or opipramol regarding all
efficacy and safety measures.75% of the patient
were classified as responders (50% reduction of
HAM-A score) in each treatment group with
60% achieving full remission. A novel study
involving 13 subjects evaluated kava‟s
potential in improving vagal control in suffers
of GAD (Watkins et al., 2001). It was observed
that significantly more patients treated with
kava showed improved BRC compared with
placebo group,reflecting a favourable effect on
reflex vagal contral of heart rate in patients
with GAD. Due to potential hazard of
hepatotoxicity, P.methysticum was withdrawn
from the European and UK markets in 2002. It
was found that the factors responsible for
hepatotoxicity included individuals hepatic
insufficiency to metabilise kavalactones
(cytochrome P-450 (CYP) 3A4 and 2D6),
incorrect cultivation (medicinal, tudie or
wichmanni varieties) being used, preparations
made using acetone or ethanolic media low in
glutathione, potentially contaminated or poorly
stored material, and use of ariel parts or root
peelings which are higher in alkaloids (Sarris et
al., 2010c). It is recommended that only peeled
roots from noble cultivers (cultivated species
that are traditionally considered safe and
therapeutic) using water soluble extraction
method is advised (Teschke et al., in press).
In a study of kava use( Av
118g/week,median duration of use=12 years) in
an Arnhem Land community in northern
territory of Australia it was found that liver
functions in users of aqueous kava at these
moderate levelsof consumption appears to be
reversible and began to return to baseline after
1–2 weeks abstinence from kava. No evidence
of irreversible liver damage has been found
(Clough et al., 2003). Kava has also been found
to cause significant drug interaction and
interactions with CYP 450 enzyme (Singh,
2005). One human pharmacokinetic trial
determined that kava caused CYP2E1
inhibition in approximately 40% (Gurley et al.,
2005). Whole kava extract (normalized to
100µm total kavalactones), caused
concentration dependent decreases in P450
activities, with significant inhibition of the
activities of CYP1A2 (56% inhibition), 2C9
(92%), 2C19 (86%), 2D6 (73%), 3A4 (78%)
and 4 A9/11(65%) following preincubation
(Mathews et al., 2002). Kava also interacts
with benzodiazepines and causes sedation
(Singh, 2005; Stevinson et al., 2002). However,
the risk-benefit ratio is highly favourable
towards kava due to respectable clinical
efficacy and relative low risk of potential liver
toxicity (1 case /million monthly doses (Bauer,
2003).
Passiflora incarnata L. (Passion flower)
It is a benzodiazepine receptor partial
agonist and causes GABA-system mediated
anxiolysis. Animal behavioural models have
shown non-sedative anxiolytic effect.In an in
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 317–336
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vivo study employing a methanol extract of
passion flower (125 mg/kg,orally) measured
anxiolytic activity in mice, using the elevated
plus-maze model,an increase in number of
entries in open arm was demonstrated (Dhawan
et al., 2001a; 2001b; 2002; Grundmann et al.,
2008; Sena et al., 2009). A 4 week RCT using
passion flower extract on patients with GAD
(n=36) showed that passion flower was as
effective as oxazepan (30 mg/day) in reducing
anxiety and it had less number of side effects
(Akhondzadek et al., 2001). In an acute study
RCT (n=60) using 500 mg of passion flower vs
placebo for presurgical anxiety (Movafegh et
al., 2008), it was demonstrated that anxiety
scores were significantly lower in the
passionflower group than in the control group
on a numerical rating scale.
Valeriana spp. (Valerian)
Felter and Lioyd demonstrated that species
of valerian officinalis and edulis have been
used in traditional American and European
medicine as a soporific and to treat various
nervous system disorders. It decreases the
degradation and simultaneously increases the
binding of GABA. Also, valerenic acid from
valerian has demonstrated GABA-A receptor
(β3 subunit) agonism and also 5-HT5a partial
agonism (Benke et al., 2009; Dietz et al., 2005;
Murphy et al., 2009; Oritz et al., 1999; Sichardt
et al., 2007; Trauner et al., 2008). A large 8
week internet based RCT (n=391) using a
valerian (6.4 valarenic acids/day) placebo, kava
(300 mg kavalactones/day) + placebo or double
placebo was conducted to determine the
efficacy in treating co morbid anxiety and
insomnia (Jacobs et al., 2005). The primary
outcome measure used in rating change in
anxiety state was STATI-State. The results
suggested that neither kava norvalerin relieved
anxiety and insomnia more than placebo. But
the design of this trial presents several potential
problems, with internet recruitment for trials
resulting in samples of questionable
representativeness, and the STATI-state having
the inadequate test-retest reliability to be a
sensitive measure of therapeutic change in
anxiety. In a systemic review and metaanalysis
of 18 RCTs (Fernandez-San Martin et al.,
2010) using Valerian vs placebo or active
controls,valerian reduced sleep latency over
placebo by only 0.70min (95% Cl-
3.44,4.83),with the standardized mean
difference between the groups measured being
stastically equivocal-0.02 (95% Cl-0.35, 0.31)
Scutellaria lateriflora L. (Skull cap)
It has a GABA-α binding affinity (Awad et
al., 2003). A double blind placebo controlled
cross over study of healhy individuals (n = 19)
revealed that skullcap dose-dependently
reduced symptoms of anxiety and tension after
acute administration compared to that with
control (Wolfson and Hoffmann, 2003). In
animal maze model test skullcap demonstrated
anxiolytic activity (Awad et al., 2003).
Melissa officinalis L. (Lemon balm)
It is shown to cause MAO-inhibition. Also
it is found to be a potent invitro inhibitor of rat
brain GABA transaminase (GABA-T) (Awad
et al., 2009; Lopez et al., 2009). An RCT with
20 participents who were given single doses of
300,600 and 900 mg of lemon balm or a
matching placebo at 7-day intervals revealed
that self rating calmness as assessed by Bond
Lader mood scales was elevated at the earliest
time points by the lowest dose, while alertness
was significantly redused at all time points
following the highest dose (Kennedy et al.,
2002). A double blind ,placebo controlled,
randomized, balanced cross over experiment
utilizing a standardized product containing
lemon balm and valerian extracts in healthy
volunteers (n=24) assessed mood and anxiety
via a DISS test (Kennedy et al., 2006).The
results demonstrated that a 600 mg dose of the
combination ameliorated the negative effects of
the DISS the level of anxiety. In a 4 week open,
multicenter study in children less than 12 years
(n=918) suffering from restlessness and
nervous dyskoimesis a combination of valerian
and lemon balm preparation (2×2 tablets /day
of 160 mg valerian root dry extract (4–5:1) and
80 mg lemon balm leaf dry extract (4-6:1) was
given.The primary symptoms of dyssomnia and
restlessness were reduced from
„moderate/severe‟ to „mild‟ or „absent „ in most
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of the children with 70.4% 0f the patients with
restlessness improving. Both parents and
investigators assessed efficacy as „very good‟
or „good‟ (65.5% and 67.7%, respectively)
(Muller and Klement, 2006).
Eschscholzia californica (DC.) Stapf. (Lemon
grass)
In 50 participants lemongrass infusion was
evaluated for hypnotic and anxiolytic activity
(Aleite et al., 1986), it was found that there was
no difference between lemon grass and
placebo.
Centella asciatica (L.) URB (Gotu Kola)
It is used in ayurvedic and traditional
pacific medicine for the tr atment of anxiety
and depression (Bone, 2003). In a double blind
placebo controlled study (Bradwejn et al.,
2000), 40 healthy participants were randomly
assigned to receive either a single 12 g orally
administered dose of gotu kola or placebo, it
was found that gotu kola significantly
attenuated peak ASR amplitude 30 and 60 min
after treatment indicating anxiolytic activity in
humans.
Withania somnifera (L.) Dona.
(Ashwagandha)
It is classified as rasayana in ayurvedic
medicine and it is used to enhance mental and
physical performance.It is widely used in the
western countries in various nervous system
disorders (Bone, 2003). In an animal study
(Bhattacharya and Muruganandam, 2003) it
was observed the adaptogenic behavior of
ashwagandha in stress –inducing procedure, via
the attenuation of stress related parameters
(cortisol levels, mental depression, sexual
dysfunction.
Bacopa monnieri (L.) Wettst. (Brahmi)
A 12 week RCT using 300 mg of brahmi
revealed that there was marked reduction in
anxiety by brahmi as compared to placebo
(Stough et al., 2001).
Ginkgo biloba L. (Maiden hair)
In a RCT using EGb 761 extract (480 mg or
240 mg per day) or placebo for 4 weeks in
adults with GAD or adjustment disorder with
anxious mood as assessed by DSM-III R using
HAM-A as the primary outcome measure and
CGI,Erlangen anxiety tension and aggression
scale (EAAS) as the secondary outcome
measure it was demonstrated that the HAM-A
total scores decresed by -14.3 ( 8.1),-12
( 9.1),and -7.8( 9.2) in the 480 mg per day
Ginkgo biloba group, the 240 mg per day
Ginkgo biloba group and the placebo group
respectively.It demonstrated specific dose
dependent anxiolysis compared with placebo in
both higher dose and lower dose group (Woelk
et al., 2007).
Crataegus spp. (Hawthorn berry/leaf)
In a RCT (Walkar et al., 2002)
administered 500 mg of hawthorn extract to
mildly hypertensive patients, there was a non
significant reduction in anxiety as compared to
placebo. A double blind ,randomized placebo
controlled trial involving adults presenting with
mild moderate GAD as assessed via DSM-III R
(n=264) were prescribes two tablets containing
fixed quantities of Crataegus oxycantha
(300mg),Eschscholtzia californica (80 mg) and
magnesium (300 mg elemental) twice daily for
3 months (Hanus et al., 2004), it was observed
that the formula was highly effective in
decreasing anxiety as compared to placebo
which was determined by HAM-A and
subjectively assessed anxiety.
CONCLUSIONS
Herbal medications in psychiatry are still
under researched. The present review looked at
various herbal preparations used in depression
and anxiety over the years. The preparations
excluding St Johns wort and kava have been
under used and need further clinical trials
including randomized double blind clinical
evidence and direct comparisons with
antidepressant drugs to help us understand their
efficacy. Most herbal medications may serve as
alternatives to traditional antidepressants in
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patients who do not tolerate them as they have
a favorable safety profile and are free from
major side effects. There is also a need for
research of herbal medication in the
management of various subtypes of depression,
bipolar disorder and anxiety disorders like post
traumatic stress disorder and obsessive
compulsive disorder. The use of these
medications in various age groups and diverse
clinical populations is warranted.
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Source of Support: Nil Conflict of Interest: None Declared
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ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
A BRIEF REVIEW ON NONI (MORINDA CITRIFOLIA L.) - A HERBAL
REMEDY FOR BETTER HEALTH
Patel Swetal1, Krishanamurthy R
2*
1M.Sc. final year project student, C.G Bhakta Institute of Biotecnology, Uka Tarsadia University, Mahuva
Road, Bardoli-394601, Surat(Dist. Surat), Gujarat, India. 2Director, C.G Bhakta Institute of Biotecnology, Uka Tarsadia University, Mahuva Road, Bardoli-394601,
Surat(Dist. Surat), Gujarat, India.
*Corresponding Author: Email: [email protected]; [email protected]; Mob: +919825349279
Received: 02/04/2013; Revised: 18/04/2013; Accepted: 22/04/2013
ABSTRACT
Noni (Morinda citrifolia L.) is an important herb of tropical regions of the world. It has been
used for over 2000 years in Polynesia. All parts of this plant are useful for many purposes. It has
more than 160 phyto - chemicals which make Noni a wonderful herbal remedy for the treatment of
many disease and disorders, also a wide range of physiochemical compounds and essential elements
which make Noni an effective health enhancer. Noni fruit juice is recently accepted as a novel food
in the European Union. Now a day Noni juice is in high demand as an alternative medicine for
various illnesses and it helps to live a long and healthy life. This review presents some active phyto-
chemicals and therapeutic effects of Noni which was proved by various scientists earlier.
KEY WORDS: Morinda citrifolia, Noni, phyto chemicals, remedy, disease, disorders, enhancer.
Review article
Cite this article:
Patel Swetal, Krishanamurthy R (2013), A BRIEF REVIEW ON NONI (MORINDA CITRIFOLIA
L.) - A HERBAL REMEDY FOR BETTER HEALTH, Global J Res. Med. Plants & Indigen. Med.,
Volume 2(5): 337–347
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 337–347
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
INTRODUCTION
Herbal remedies refer to the use of plants
for the promotion of healing and maintenance
of health. Till date about 80 % of people in
developing countries still relays on traditional
medicine based largely on species of plants and
animals for their primary health care. Herbal
medicines are currently in demand and their
increasing day by day (Von Reis, 1977).
Noni, with the botanical name Morinda
Citrifolia L., is a medicinal plant which is used
as a natural nutritional food supplement
worldwide for centuries. Morton (1990)
reported that the fruit of Noni plant have a
history of use in the pharmacopoeias of pacific
islands and south East Asia. Noni is one of the
important traditional folk medicinal plants that
have been used for over 2000 years in
Polynesia. The Polynesians utilized the whole
Noni plant for herbal remedies. The species of
Morinda especially M. citrifolia has been
reported to have a broad range of health
benefits for cancer, infections, arthritis, asthma,
hypertension, and pain (Whistler, 1992). The
leaves, seeds, bark, fruits and roots of Noni
have been used in various topical remedies in
South Pacific Islands and South East Asia
(Wang et al., 2002; Fygh-Berman, 2003). The
various therapeutic benefits of Noni are due to
the enriched phyto constituents. The high
therapeutic profile and safety potential of Noni
has made it a popular health enhancer and food
supplement worldwide.
PLANT DESCRIPTION
Morinda citrifolia L., is a small tree with a
height of 3–10 meter. It has an abundant long
& broad elliptic leaves (5–17 cm length &10–
40 cm width). Flowers are small, tubular and
white in colour. They are arranged in group and
inserted in one peduncle. The petiole leave ring
like marks on stalks and the corolla is greenish
white. (Morton, 1992; Elkins, 1998; Dixon et
al., 1999; Ross, 2001; Cardon, 2003). Fruits of
Noni plant are oval and have an embossed
appearance. Fruits have a range of colour from
dark green to yellow with their different
maturity stages. (Morton, 1992; Dixon et al.,
1999). A mature fruit appear almost white in
colour and is covered with a small reddish
brown bud containing the seeds. The unripe
fruit is odourless but the ripe fruit has a strong
smell of butyric acid. The seeds are triangular
in shape and radish brown in colour (Dittmar,
1993).
Chemical constituents
About 160 phyto - chemical compounds
have been already identified in the Noni plant,
and the major micronutrients are Phenolics
compounds, organic acids and alkaloids (Wang
and Su, 2001). Several classes of compounds
have been isolated from M. citrifolia L.,
including amino acids, anthraquinones,
coumarins, fatty acids, flavonoids, iridoids,
lignans and polysaccharides (Chan-Blan-co et
al., 2006). However, chemical composition
differs significantly according to the part of the
plant. The complete physico - chemical
composition of the fruit has not yet been
reported, and only partial information is
available on M. citrifolia L. juice. The fruit
contains 90% water, and the main components
of the dry matter appear to be soluble solids,
dietary fibre and proteins (Chunhieng, 2003).
Xeronine system
Noni fruit contains a natural precursor for
Xeronine that Heinicke named Proxeronine.
Proxeronine is converted to the alkaloid,
Xeronine, in the body by an enzyme
Proxeroninase. A hypothesis is that Xeronine is
able to modify the molecular structure of
proteins. Thus Xeronine has a wide range of
biological activities. When a protein such as an
enzyme, receptor, or signal transducer is not in
the appropriate conformation, it will not work
properly. Xeronine will interact with the
protein and make it fold in to its proper
conformation. The result is a properly
functioning protein (Heinicke, 1985).
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Table: 1 Some active bio chemical component of Noni juice & leaf powder
Characteristics
Fruit juice Leaf powder
Chunhieng
(2003)
Shovic and
Whisler (2001)
European
commission (2002)
(Leung et al.,
1972)
Protein 2.5% 0.4g/100g 0.2–0.5% 1 g/100g
Lipid 0.30g/100g 0.1–0.2% − 0.2 g/100g
Glucose 11.9 ± 0.2 g/l − 3.0–4.0% −
Fructose 8.2 ± 0.2 g/l − 3.0–4.0% −
Potassium 3900 mg/l 188 mg/100g 30–150 mg/100g −
Sodium 214 mg/l 21 mg/100g 15–40 mg/100g −
Magnesium 14 mg/l 14.5 mg/100g 3–12 mg/100g −
Calcium 28 mg/l 41.7 mg/100g 20–25 mg/100g 58 g/100g
Vitamin C − 155 mg/100g 3–25 mg/100g 50 g/100g
Table: 2 Chemical constituent, there place and its chemical nature in different plant part
recognize by various authors:
Plant part
(place)
Chemical constituent Structure Reference
Flower 2-methyl-4-hydroxy-5,7-
dimethoxyanthraquinone
4-O-ß-D-glucopyranosyl
(1, 4)-a-L-rhamnopyranoside
Anthraquinone
Glycosides
Sang et al., (2002)
5,8-dimethyl-apigenin
4'0-O-ß-D-galactopyranoside
Flavonoids Sang et al., (2002), Elkins
(1998)
Fruit
2,6-di-O-(ß-D-glucopyranosyl
1-O-octanoylß- D
glucopyranose
Fatty acid ester Dittmar (1993)
6-O-(ß-D-glucopyranosyl-1-O-
octanoyl-ß-D glucopyranose
glycosides Wang et al., (1999)
Ascorbic acid Acid Liu et al., (2001)
Asperulosidic acid Flavonoids Morton (1992), Elkins (1998)
Aspuruloside tetraacetate Flavonoids Wang et al., (1999), Liu et
al., (2001) Cardon (2003)
Caproic acid Acid Elkins (1998)
Caprylic acid Acid Sang et al., (2002), Dittmar
(1993),Wang et al., (2002),
Elkins (1998), Liu et al.,
(2001)
Ethyl acetate Ester Elkins (1998)
Ethyl caprylate Ester Solomon (1999b),Elkins
(1998), Dittmar (1993), Wang
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et al.,(2002), Cardon (2003),
Liu et al.,(2001), Srivastava
& Singh (1993)
Ethyl caproate Ester Dittmar (1993)
Hexanoic acid Acid Dittmar (1993)
Quercetin
3-O-a-L-rhamnopyranosyl-
(1-6)-ß-D-glucopyranoside
Flavonoide Sang et al., (2002), Cardon
(2003), Wang & Su (2001),
Farine et al., (1996)
Leaves Quercetin
3-O-a-L-rhamnopyranosyl-
(1-6)-ß-D-glucopyranoside
Flavonoids Sang et al., (2002)
Alanine Amino acid Sang et al., (2002), Cardon
(2003), Srivastava & Singh
(1993)
Serine Amino acid Dittmar (1993), Elkins (1998)
Threonine Amino acid Dittmar (1993), Elkins (1998)
Tryptophan Amino acid Dittmar (1993), Elkins (1998)
Tyrosine Amino acid Dittmar (1993), Elkins (1998)
Urosolic acid Triterpenoids
and sterols
Sang et al., (2002), Elkins
(1998), Wang et al., (2002),
Cardon (2003)
Valine Amino acid Dittmar (1993), Elkins
(1998)
Arginine Amino acid Dittmar (1993)
Aspartic acid Amino acid Dittmar (1993)
ß-sitosterol
Sterols Sang et al., (2002), Wang et
al., (2002), Chunhieng (2003)
Citrifolinoside B Iridois Sang et al., (2002)
Cysteine Amino acid Elkins (1998)
Glutamic acid Amino acid Dittmar (1993)
Glycine Amino acid Dittmar (1993), Elkins (1998)
Histidine Amino acid Dittmar (1993), Elkins (1998)
Isolucine Amino acid Dittmar (1993), Elkins (1998)
Leucine Amino acid Dittmar (1993), Elkins (1998)
Phenylalanine Amino acid Dittmar (1993), Elkins (1998)
Methionine Amino acid Dittmar (1993), Elkins (1998)
Proline Amino acid Dittmar (1993), Elkins (1998)
Kaempferolm 3-O-ß-D-
glucopyranosyl-(1-2)- a-
Lrhamnopyranosyl-(1-6)- ß-D-
galactopyranoside
Chlorophyll
derivatives
Sang et al., (2002)
Quercetin 3-O-ß- D-
glucopyranoside
Flavonoids Sang et al., (2002)
Stem 2-hydroxyanthraquinone Anthraquinone Siddiqui et al., (2006)
2-methoxyanthraquinone Anthraquinone Siddiqui et al., (2006)
Morindicininone Anthraquinone Siddiqui et al., (2006)
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 337–347
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Seeds Ricinoleic acid Acid Solomon (1999b)
Root
8-hydroxy-8-methoxy-2-
methyl-anthraquinone
Anthraquinone Cardon (2003),
Solomon (1999b)
rubichloric acid Acid Elkins (1998)
1,3-dihydroxy-6-
methylAnthraquinone
Anthraquinone Morton (1992)
Morenone 1 Anthraquinone Solomon (1999b)
Morenone 2 Anthraquinone Solomon (1999b)
Ruberythric acid Acid Cardon (2003)
Rubiadin Anthraquinone Elkins (1998), Cardon (2003),
Inoue et al., (1981), Ross
(2001)
Root bark Chlororubin Chlorophyll
derivatives
Dittmar (1993)
Hexose Saccharide Dittmar (1993)
Morindadiol Anthaquinone Dittmar (1993)
Morindanidrine Anthraquinone Dittmar (1993)
Morindine Anthraquinone Elkins (1998), Cardon (2003)
Pentose Saccharides Dittmar (1993)
Physcion Anthraquinone Solomon (1999b)
Rubiadin monomethyl ether Phenol Dittmar (1993)
Soranjidiol Anthraquinone Dittmar (1993), Elkins
(1998), Ross (2001)
Trioxymethyl anthraquinone
monoethyl ether
Anthraquinone Dittmar (1993)
Hard wood Physcion 8-O-a-L
arabinopyranosyl-
(1-3)-ß-Dgalactopyranosyl-(1-
6)- ß -D-galactopyranoside
Anthraquinone
Glycoside
Wang & Su (2001), Wang et
al., (2002)
Root,
Hard
wood,
Root bark
Morindone Anthraquinone Sang et al., (2002), Dittmar
(1993), Wang et al., (2002),
Ross (2001)
Root,
Hard
wood,
Seeds
Damnacanthal Anthraquinone Sang et al., (2002), Cardon
(2003)
Root, Root
bark, Fruit
Alizarin Anthraquinone Elkins (1998), Dittmar
(1993), Wang et al., (2002),
Cardon (2003), Ross (2001),
Dittmar (1993)
Plant
2-methyl-3,5,6
Trihydroxyanthraquinone
Anthraquinone Cardon (2003)
Scopoletin Miscellaneous
compound
Wang et al., (2002), Farine et
al., (1996)
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2-methyl-3,5,6-
trihydroxyanthraquinone
6-O-ß-D-xylopyranosyl-(1-6)-
ß-D-glucopyranoside
Anthraquinone
glycoside
Cardon (2003), Inoue et al.,
(1981)
3-hydroxymorindone Anthaquinone Cardon (2003), Inoue et al.,
(1981)
3-hydroxymorindone
6-O-ß-D-xylopyranosyl- (1-6)-
ß-Dglucopyranoside
Anthraquinone
Glycoside
Cardon (2003), Inoue et al.,
(1981)
5,6-dihydorxylucidin 3-O-ß-D-
xylopyranosyl-(1-6)- ß-D –
glucopyranoside
Anthraquinone
glycoside
Cardon (2003), Inoue et al.,
(1981)
5,6-dihydroxylucidin Anthraquinone Cardon (2003), Inoue et al.,
(1981)
Aucubin Anthraquinone Elkins (1998), Wang et al.,
(2002)
Linoleic acid Acid Wang et al., (2002)
Lucidin Anthraquinone Cardon (2003), Inoue et al.,
(1981)
Lucidin 3-O-ß-Dxylopyranosyl-
(1-6)-ß-Dglucopyranoside
Anthraquinone
glycosides
Cardon (2003)
Some selective medicinal, biological and
therapeutic activity of noni
The Polynesians utilized the whole Noni
plant for herbal remedies. The fruit juice is in
high demand in alternative medicine for
different kind of illnesses such as arthritis,
diabetes, high blood pressure, muscle aches and
pains, menstrual irregularities, headache, heart
disease, AIDS, cancers, gastric ulcer, sprains,
mental depression, senility, poor digestion,
arteriosclerosis, blood vessel problems, and
drug addiction. Scientific evidence of the
benefits of the Noni fruit juice is limited but
there are some subjective evidences for
successful treatment of colds and influenza
(Solomon, 1999a). It is reported to have
antibacterial, anti-fungal, analgesic,
hypotensive, anti inflammatory and Immuno-
stimulatory effects (McClatchey W., 2002 a &
b; Wang et al., 2002; Mathivanan et al., 2005).
Noni has a broad range of therapeutic effects
such as analgesic, anti-inflammatory,
antihypertensive, immune enhancing,
anticancer, antibacterial, antiviral, antifungal,
antistress, antituberculous, antiprotozoal,
antioxidant, and also sedative properties, also
Noni is effective in cough, nausea, enlarged
spleen, joint disorders such as gout and
arthritis, senility, poor digestion, arthrosclerosis
and drug addiction. These beneficial effects of
Noni are strongly documented and well
authenticated by valid scientific literature
evidences. Noni also has a strong cancer
preventive effect (Murugesh, 2007).
a. Anti- cancer activity
Damnacanthal, an anthraquinone extracted
from Noni induces normal morphology in a
particular type of cell found in human
neoplasias (K-ras-NKR cells) that multiply
uncontrollably and are highly malignant
(Hiramatsu et al., 1993; Hirazumi et al.,
1996; Hirazumi & Furusawa, 1999).
b. Anti HIV activity
A compound isolated from Noni roots
named 1 - methoxy - 2 – formyl – 3 -
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 337–347
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
hydroxyanthraquinone suppresses the
cytopathic effect of HIV infected MT-4
cells without inhibiting cell growth
(Umezawa et al., 1992).
c. Anti tubercular activity
At the International Chemical Congress of
the Pacific Basin Societies meeting in
Honolulu, Saludes and colleagues from the
Philippines reported that Noni Kills
Mycobacterium tuberculosis. A
concentration of extracts from Noni leaves
killed 89% of the bacteria in a test tube,
almost as effectively as the leading anti-TB
drug Rifampicin, which has an inhibitory
rate of 97% at the same concentration.
d. Anti tumour activity
TNJ showed dose-dependent cytotoxicity
on cultured cancer cells by inducing cancer
cell necrosis at high doses and apoptosis at
lower doses. Synergistic effects of TNJ
with known anticancer drugs have been
found (Wang et al., 2002).
e. Immunological activity
An alcohol extract of Noni fruit at various
concentrations inhibits the production of
tumour necrosis factor-alpha (TNF-α),
which is an endogenous tumour promoter.
Therefore, the alcohol extract may inhibit
the tumour promoting effect of TNF-α
(Ashina et al., 1994). Noni is also capable
of stimulating the release of several
mediators from murine effector cells,
including TNF-α, interleukin-1 beta (IL-1),
IL-10, IL-12, interferon-gamma (IFN- α
and nitric oxide (NO) (Himzumi et al.,
1990).
f. Anti oxidant activity
The SAR scavenging activity of Noni juice
was shown to be 2.8 times higher than that
of vitamin C, 1.4 times that of pycnogenol
(PYC) and almost the same magnitude as
that of grape seed powder (Wang & Su,
2001).
g. Analgesic activity
The results showed that rats fed 10% or
20% Noni juice had greater pain tolerance
(162% or 212%, respectively) compared
with the placebo group (Wang et et al.,
2002). Noni root extract (1600 mg/kg)
showed significant analgesic activity in the
animals, similar to the effect of morphine
(75% and 81%) protection using Noni
extract and morphine, respectively), and it
also proved to be non-toxic (Younos et al.,
1990).
h. Anti inflammatory activity
Commercial M. citrifolia L. juice
selectively inhibits cyclo oxygenase
enzymes (COX-1 and COX-2) involved in
breast, colon and lung cancer and also has
anti-inflammatory activity (Su et al., 2001).
The ability of Noni juice to inhibit these
enzymes was compared to that of
traditional commercial non-steroidal
inflammatory drugs, such as aspirin,
Indomethacin and Celebrex.
i. Anti lithiatic effect
M. citrifolia. L has the anti Lithiatic effect
on Ethylene Glycol induced Lithiasis in
male albino rats. This observation provided
the basis for considering Noni for inhibiting
stone formation induced by ethylene glycol
(Murugesh and Christina, 2007).
j. Anthelmintic activity
An ethanol extract of tender Noni leaves
was found to induce paralysis and death in
the human parasitic nematode Ascaris
Lumbricoides within a day (Raj, 1975).
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 337–347
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k. Hypotensive activity
A Hawaiian physician reported Noni, fruit
juice to have a diuretic effect (Youngken et
al., 1960).
A hot water extract of Noni roots lowered
the blood pressure of an anesthetized dog
(Youngken et al., 1958, Davison C, 1927).
A Hawaiian physician reported that Noni
fruit juice had a diuretic effect (Asahina AY
et al., 1994).
l. Anti fungal activity
Recent research has demonstrated that it
contains a water-soluble component or
components that interfere with the
morphological conversion of Candida
albicans and may have potential therapeutic
value with regard to candidiasis (Banerjee
et al., 2006; Usha et al., 2010). Other
studies showed that methanol extract of the
dried fruit exhibited maximum percentage
of inhibition against Trichophyton
mentagrophytes (79.3%), while
approximately 50% activity was recorded
against Penicillium, Fusarium and Rhizopus
species (Jainkittivong et al., 2009).
m. Anti bacterial activity
It has been reported that M. citrifolia L.
inhibits the growth of certain bacteria, such
as Staphylococcus aureus, Pseudomonas
aeruginosa, Proteus morgaii, Bacillus
subtilis, Escherichia coli, Helicobacter
pylori, Salmonella and Shigella (Atkinson,
1956). Anti-microbial effect observed may
be due to the presence of phenolic
compounds such as acubin, l-asperuloside,
alizarin, scopoletin and other
anthraquinones. Another study showed that
an acetonitrile extract of the dried fruit
inhibits the growth of Pseudomonas
aeruginosa, Bacillus subtilis, Escherichia
coli, and Streptococcus pyrogene (Locher et
al., 1995).
n. Anxiolytic activity
Recent research has demonstrated the
effects of Noni fruit on preventing anxiety
disorders, affecting an estimated 25% of the
adult population at some point during their
lifetime (Kjernised and Bleau, 2004).
o. Cardio vascular activity
Recent research has demonstrated the
ability of Noni fruit to prevent
arteriosclerosis, a disease related to the
oxidation of low density lipoproteins
(LDLs). This beneficial effect could be due
to presence of lignans (Kamiya et al.,
2004).
p. Esrogenic activity
Recent research has demonstrated the
effects of Noni fruit on preventing anxiety
disorders, affecting an estimated 25% of the
adult population at some point during their
lifetime (Kjernised and Bleau, 2004).
CONCLUSION
Noni with a botanical name Morinda
citrifolia has a long history of widespread use
as a food in tropical regions from Indonesia to
the Hawaiian Islands, and it is used as an herbal
remedy for multiple diseases. It carry several
vitamins, minerals, micro and macro nutrients
that help the body in various ways from cellular
level to organ level. It also carries several phyto
- chemicals which help to prevent severe
infection and many diseases. It has been proved
by various researchers that, drinking TNJ was
beneficial for the prevention of heart, lung, and
brain diseases as well as delaying the ageing
processing, and maintaining overall good
health so now a day a wide range of
therapeutic effects of Noni make it a popular
herbal medicine for better health.
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 337–347
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
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Source of Support: Nil Conflict of Interest: None Declared
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 348–356
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
DEVELOPMENT OF RANDOM AMPLIFIED
POLYMORPHIC DNA MARKERS FOR AUTHENTICATION OF
RIVEA HYPOCRATERIFORMIS (DESR.) CHOISY
Borkar Sneha D1, Naik Raghavendra
2, Harisha C R
3, Acharya R N
4
1,2P G Scholar, Dravyaguna Department, IPGT&RA, Gujarat Ayurved University, Jamnagar, Gujarat, INDIA
3Head, Pharmacognosy Laboratory, IPGT&RA, Gujarat Ayurved University, Jamnagar, Gujarat, INDIA
4Associate Professor, Dravyaguna Department, IPGT&RA, Gujarat Ayurved University, Jamnagar, Gujarat,
India
*Corresponding Author: E-mail: [email protected]
Received: 26/03/2013; Revised: 17/04/2013; Accepted: 20/04/2013
ABSTRACT
Rivea hypocrateriformis (Convolvulaceae) is consumed as a leafy vegetable and also reported for
its ethno-medicinal uses in cough, headache, skin disease etc. The Pharmacognostic study of its leaf
and molecular characterization of the plant by Random Amplified Polymorphic DNA (RAPD)
markers was studied following standard parameters. Genomic isolation of DNA from fresh leaves
was amplified by RAPD markers. The unique bands obtained in Polymerase Chain Reaction (PCR)
amplification were clearly discriminated having, many bright and light bands indicating the
genuinity of the plant. The diagnostic characters of its leaf were presence of paracytic stomata, oil
globule, trichomes, rosette crystals and tannin. The findings of this study may provide useful
information with regards to its DNA, pharmacognostic and taxonomic species identification.
KEYWORDS: DNA finger prints, Fanji, Leaf drug, Pharmacognosy, Rivea hypocrateriformis,
Stomatal index
Research article
Cite this article:
Borkar S D, Naik R, Harisha C R, Acharya R N (2013), DEVELOPMENT OF RANDOM AMPLIFIED
POLYMORPHIC DNA MARKERS FOR AUTHENTIFICATION OF RIVEA
HYPOCRATERIFORMIS (DESR.) CHOISY, Global J Res. Med. Plants & Indigen. Med.,
Volume 2(5): 348–356
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 348–356
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
INTRODUCTION
The use of plants as source of food is as old
as humanity. Classical texts of Ayurveda
recommend medicine as well as dietetic items
(ahara dravyas) for the management of
different disease condition like Jwara (fever),
Atisara (diarrhea) Shwasa (asthama) etc.
Ayurveda delineates these types of ahara
dravyas under the heading shakavarga (group
of vegetables). Fanji, botanically identified as
Rivea hypocrateriformis (Desr.) Choisy
(Convolvulaceae), is used as leafy vegetable
and for medicinal purposes. Its leaves are taken
internally, to relieve cough and headache (A.
Sarvalingam et al., 2011); as food supplement
(Tribhuban Panda et al., 2007); in rheumatism
and skin disease, as an analgesic, anti
inflammatory (B. Swathy et al., 2010); piles
(H.M. Patil et al., 2006) and whole plant
powder is added to milk and taken in sexual
weakness (S.Y. Kamble et al., 2010).
Random amplified polymorphic DNA
(RAPD) analysis is powerful and convenient
molecular marker system, widely used for the
genetic mapping, taxonomic and polygenic
studies of many plants. Review of literature
reveals that leaves of Rivea hypocrateriformis
have not been studied, in detail, for molecular
characterization through RAPD analysis except
some pharmacognostical characters of its leaf.
(Rajiv Kukkar et al., 2011) Hence, the present
study was undertaken to establish certain
botanical standards for identification and
standardization of R. hypocrateriformis leaf.
MATERIALS AND METHODS:
Collection and preservation of the sample:
Leaves of Rivea hypocrateriformis, were
collected from its natural habitat, Rakha khatia
forest area, Jamnagar, Gujarat, during October
2012 and identified with the help of Forest
flora of Gujarat state (R I Patel, 1984). A
sample specimen was authenticated by expert
taxonomist and deposited to institutes
pharmacognosy museum (SPECIMEN NO-
PHM 6063/21/09/2012) for future references.
The leaves were washed, shade dried,
powdered, sieved through 80 mesh and
preserved in an air-tight glass vessel. For
microscopical evaluation, fresh samples were
preserved in a solution prepared from 70%
ethyl alcohol: glacial acetic acid: formalin
(AAF) in the ratio of 90:5:5 (Johnson
Alexander Donald, 1940). Fresh leaves of
Rivea hypocrateriformis were utilized for
RAPD analysis.
Pharmacognostic studies:
Morphological characters were studied by
observing the leaves as such and also with the
help of the dissecting microscope. For detailed
microscopical observation, free hand thin
transverse sections passing through the midrib
were taken. Observed sections were cleared
with chloral hydrate and observed as such for
the presence of any crystals, then were stained
with Phloroglucinol and Hydrochloric acid
(HCl) to notice the lignified elements like
fibers, vessels etc. of the meristele and other
parts (Khandelwal K.R., 2008). Photographs of
the section were taken with the help of Canon
digital camera attached to Zeiss microscope.
Powder characters were studied as per the
guidelines of Ayurvedic Pharmacopoeia of
India, (Anonymous, 1999). The histo-chemical
tests were carried out according to the standard
guidelines. (Krishnamurty K. V, 1988).
Surface study and micrometry
Surface study of epidermis was carried out
to determine type and distribution of stomata,
epidermal cell and trichomes. Quantitative
microscopy was carried out to determine
epidermal cell number, stomatal number,
stomatal index and size of the stomata etc.
Mean values are taken by five successive
readings. (Wallis, 1985).
The stomatal index was found by using
following formula
I = S x 100
E + S
(I = stomatal index, S = no. of stomata per unit
area, E = no. of epidermal cells in the same unit
area.)
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Molecular characterization (DNA
fingerprints)
Fresh leaves were used in molecular
characterization and DNA fingerprints were
obtained by standard and most convenient
RAPD method. The RAPD reaction was
performed following standard procedures
(Baum BR, Mechanda S., 2001) at Aristogene
Biosciences Pvt. Ltd, Bangalore.
DNA isolation:
Young leaves were selected, cut into small
pieces without cutting the veins. They were
washed with distilled water and ethanol. Frozen
with dry ice and crushed. To that, 2 ml of plant
DNA extraction buffer was added. The samples
were ground thoroughly, transferred into
centrifuge tube and added 10 ml plant DNA
extraction buffer. 50 µl of BME added, to each
tube, mixed well. Incubated at 65ºC for 1 hour
with intermittent mixing. Centrifuged for 15
minutes at 10 K (10000). Supernatant was
transferred carefully into fresh tube and added
equal volume of chloroform and mixed well.
Centrifuged for 15 minutes at 10 K (10000).
Aqueous layer carefully pipetted into fresh tube
and precipitated with isopropanol. DNA pellet
suspended in 300µl of TE and subjected to
column purification.
Column purification
Silica spin columns and buffers were from
Qiagen
The column was placed in collection tube,
400 µl of equilibration buffer was added to the
column and centrifuged at 10000 rpm for 1min.
Collected buffer was discarded. 400 µl of
equilibration buffer was added to the DNA
samples, mixed and loaded into the column
(This step was repeated till the DNA sample
was completed). Flow through was collected.
500 µl of wash buffer 1 was added, centrifuged
at 10000 rpm for 1minute and buffer was
collected. 500 µl of wash buffer 2 was added,
centrifuged at 10000 rpm for 1minute and
buffer was collected. The empty column was
centrifuged with collection tube to completely
remove the wash buffer for 2 minute. 50 µl of
elution buffer was added to the column placed
in new collection tube. Incubated at room
temperature for 2 minutes and centrifuge at
10000 rpm for 1minute and eluted sample was
saved (elution 1). Previous step was repeated
(elution 2). Quantization of eluted DNA
samples was done by loading into the agarose
gel (Table 1–3).
Table 1: RAPD PCR
Sequences of primers used:
OPA-02 TGCCGAGCTG
OPB-10 CTGCTGGGAC
OPC-06 GAACGGACTC
Table 2: Reactions were set up with PCR master mix and respective Random primer. OPA-02 OPC-06 OPB-10 Notes
Double Distilled
water
18 µl 18 µl 18 µl
2X PCR master mix 20 µl 20 µl 20 µl 1X Contains 100µM each of dATP,
dGTP, dCTP and dTTP. Assay
buffer with 15mM MgCl2,
3U/reaction Taq Polymerase.
DNA sample 1 µl 1 µl 1 µl 10 pM used for each reaction
Random primer 1 µl 1 µl 1 µl
Total Volume 40 µl 40 µl 40 µl
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Table 3: PCR Conditions:
Temperature Time No. of cycles
94˚C 2 minutes 1
94˚C 30 seconds 40
45˚C 1 minute
72˚C 1 minute 30 seconds
72˚C 7 minutes 1
RESULT AND DISCUSSION:
Morphological study:
Leaves were alternate, simple, ex-stipulate, orbicular, dark green above light greenish blue below, appressedly silky hairy beneath when young, petiolate, petiole 2–6.3cm long, silky, lamina 6–7 × 6–6.5 cm, contained 6 pairs of nerves, nerves and midrib surface were light purplish, margin entire, base chordate, apex obtuse, dark purple glands were present at the base where lamina is attached to the petiole. (PLATE A-1, 2)
Transverse section of petiole
T S showed an outermost single layer of epidermis covered with cuticle. Unicellular warty trichomes were present on the epidermis. A single layer of parenchymatous hypodermis present beneath the epidermis consisted of the rosette crystals at places extended upto 2–3 layers followed by chollenchymatous 4–5 layers. In cortical region some parenchyma cells consisted of yellowish brown content was observed, which may be tannin. Meristele with bi-collateral vascular bundles was present at the centre portion. Each vascular bundle consisted of phloem towards lower epidermis and xylem towards center region. The vascular bundles were separated by uniseriate to biseriate medullary rays. Xylem consisted of xylem parenchyma and fibers. Some of the parenchyma cells also consisted of oil globules. The xylem parenchyma cells and also medullary rays were filled with tannin material. (PLATE –A 3, 4)
Transverse section of leaf
Leaf was dorsi-ventral, T.S. of the leaf through mid rib showed distinguished upper
palisade and lower spongy parenchyma and centrally located large vascular bundles. Upper and lower epidermis single layered, barrel shaped and consisted of compactly arranged cells with thick cuticle. Some of the epidermal cells on both the epidermis were with unicellular warty trichomes. Stomata were present more on lower epidermis as compared to upper epidermis. (PLATE – A 9, 10, 11)
Mesophyll differentiated into upper palisade and lower spongy parenchyma. Palisade parenchyma were elongated, 2–3 layered, compactly arranged, heavily filled with chloroplast pigments. Some of the palisade parenchyma cells consisted of rosette crystals of calcium oxalate. Spongy parenchyma present at side of lower epidermis, rounded to oval in shape with large intercellular spaces filled with chlorophyll pigment and some rosette crystals. (PLATE – A 7, 8)
Through midrib
On either side of upper and lower epidermis 1–2 layers of collenchyma cells, thick compactly arranged give mechanical support to ground tissue. Underneath the collenchyma, parenchymatous ground tissue compactly arranged without any intercellular spaces, consisted of large number of rosette crystals of calcium oxalate and tannin material towards the lower side of epidermis. Large vascular bundle situated at centre of the section which was collateral and closed in type, surrounded by parenchymatous sheath. Phloem was towards lower epidermis with some sieve elements. Xylem was towards upper epidermis, meta-xylem facing phloem and proto-xylem towards centre. Xylem consisted of some xylem parenchyma cells with fibres (PLATE– A 5, 6).
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PLATE – A: Pharmacognostic characters of Rivea hypocrateriformis (Desr.) Choisy
1.Plant in natural Habitat
2.Leaf measurements
3.T.S. of petiole (unstained)
4. T.S. of petiole (stained)
5.T.S. through midrib
(unstained)
6.T.S. through midrib
(stained)
7.Pallisade with rosette crystal
8.Pallisade 2-3 layered
9.Stomata upper epidermis
10.Stomata lower epidermis
11.Stomata lower epidermis
with measurments
12.Simple warty trichomes
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13.Simple fibers
14.Trichome thick walled
15.Paracytic stomata
16.Rosette crystals
17.Spiral vessels
18.Annular vessels
19.Tannin containg cell
20. Histo – chemical test
Table 4: Quantitative microscopic analysis of leaf
Sr. No. Characters Results
Upper Epidermis Lower epidermis
1 Stomata length 9.0 µm 9.0µm
2 Stomata breadth 7.0 µm 7.0µm
3 Epidermal cell 8 × 7 µm2 9 × 9 µm
2
4 Stomatal circumference 21 µm 2 24 µm
2
5 Epidermal cell circumference 24 µm2 26 µm
2
6 No. of stomata 6 9
7 No. of epidermal cells 36 26
8 Stomatal index 15 24
9 Palisade ratio 2–3 Nil
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Table 5: Histo – chemical test of leaf of Rivea hypocrateriformis
Sr. no Reagent Observation Characteristics Result
1. Phloroglucinol+
Concentrated HCl
Red Lignified cells ++
2. Iodine Blue Starch grains −
3. Phloroglucinol+
Concentrated HCl
Dissolved Calcium oxalate
crystals
++
4. Fecl3 solution Dark blue to
black
Tannin cells ++
5. Sudan III Red Oil globules ++
PLATE - B
Surface study & micrometry
The stomatal length and breadth of upper
and lower epidermis measured 9.0 µm and
7.0 µm respectively. Epidermal cells of upper
epidermis measured about 8 × 7 µm2
and that of
lower epidermis measured 9 × 9 µm
2. Stomatal
circumference measured 21 µm2 and 24 µm
2
where as epidermal cell circumference
measured 24 µm2
and 26 µm2
of upper and
lower epidermises respectively. The study
showed number of stomata 6 & 9, numbers of
epidermal cells 36 & 26, and stomatal index 15
& 24 of upper and lower epidermis
respectively. The palisade ratio of upper
epidermis was 2–3. Upper and lower epidermis
of leaf showed paracytic stomata, oil globule
and epidermal cells (Table 4). The stomatal
index number and palisade ratio are constant in
all geographical area but this may change in
extreme environmental condition. Micrometric
evaluation is the only way to specify the
specific species.
Histo – chemical test
To confirm the presence and absence of the
chemical constituents the material was
subjected to various tests. Lignified cells,
calcium oxalate crystals, tannin, oil globule
were present in the leaf.
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Powder microscopy
Organoleptic characters shows greenish
colour with characteristic odour, bitter taste,
coarse in touch. Diagnostic character of leaf
powder showed simple trichomes, warty
trichomes and stomata (Paracytic) of both the
epidermis. Rosette crystals of calcium oxalate,
tannin content of mesophyll. Spiral and annular
vessels of vascular bundles. Simple fibers and
fragments of spongy parenchyma. (PLATE –
12–19)
DNA finger printing:
All the primers gave good band pattern.
Very prominent band at ~1kb and ~0.65 kb was
obtained with OPB-10 and OPA-02
respectively (Plate – B).
CONCLUSION
The unique bands obtained in Polymerase
Chain Reaction (PCR) amplification are clearly
discriminated having, many bright and light
bands indicating the genuinity of the plant
Rivea hypocrateriformis (Desr.) Choisy. The
leaf of R. hypocrateriformis can be identified
on the basis of key microscopical and
histochemical tests which showed the presence
of characters like paracytic stomata, oil
globule, trichomes, rosette crystals and tannin.
The micrometric value such as the stomatal
index (15 upper & 24 lower); stomatal length
and breadth of upper and lower epidermis
(9.0 µm × 7.0 µm); palisade ratio of upper
(2−3) and absence in lower epidermis. The
observed DNA finger prints and
pharmacognostical characters like stomatal
type, statistical data of palisade ratio, stomatal
index, stomatal number may be useful to
establish the botanical standards for
identification and standardization of Rivea
hypocrateriformis leaf.
ACKNOWLEDGEMENT
The authors like to acknowledge the
administrative authorities of the institute IPGT
& RA, Jamnagar for providing facilities during
work. Authors also express their sincere thanks
to Dr. Sudha, Director, Aristogene Biosciences
Pvt Ltd, Bangalore for their co-operation for
DNA RAPD study of the plant.
REFERENCES
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Pharmacopoeia of India. Govt. of India
publication, New Delhi. 1st edition. 1:
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their DNA fingerprints.
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for analgesics and anti inflammatory
activities. IJBPR. 1(1):7–12.
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Bhaishajya Ratnavali, Chaukhambha
Prakashana, Varanasi, pp no.201, 202,
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knowledge system of tribals of Nandurbar
district Maharashtra. Indian journal of
traditional Knowledge. 5(3):327–330.
Johnson Alexander Donald (1940). Plant Micro
technique. Macgrow Hill Book Company,
New York, London. pp. 105.
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Khandelwal K.R. (2008), Practical
Pharmacognosy Techniques and
Experiments, 19th
Ed, Nirali Prakashan;
2008. pp no. 15– 18.
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plant histochemistry, Vishwanadhan Pvt
Limited, Madras, pp.1– 77.
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Forest department, Gujarat state, Baroda.
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edition. pp. 218.
Rajiv Kukkar, Mona kukkar, A.K.Saluja (2011)
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hypocrateriformis inventi rapid: planta
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Sawant, S G Pawar, E A Singh (2010).
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Sustainable food habits of the hill
dwelling kandha tribe in Kalahandi
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traditional medicine. 6(1):103–105.
Wallis TE (1985). Textbook of Pharmacognosy.
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Source of Support: Nil Conflict of Interest: None Declared
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Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
ANTIMICROBIAL EVALUATION OF CROTON ROXBURGHII BALAK.
(EUPHORBIACEAE) STEM BARK
Patel Esha1*, Padiya RH
2, Acharya RN
3
1Ph.D. Scholar, Dept. of Dravyaguna, Institute for Postgraduate Teaching and Research in Ayurveda, Gujarat
Ayurved University, Jamnagar, Gujarat – 361 008, India. 2Ph.D. Scholar, Dept. of Dravyaguna, Institute for Postgraduate Teaching and Research in Ayurveda, Gujarat
Ayurved University, Jamnagar, Gujarat – 361 008, India. 3Associate Professor, Dept. of Dravyaguna, Institute for Postgraduate Teaching and Research in Ayurveda,
Gujarat Ayurved University, Jamnagar, Gujarat – 361 008, India.
*Corresponding Author: Email: [email protected]
Received: 26/02/2013; Revised: 10/04/2013; Accepted: 15/04/2013
ABSTRACT
Antibacterial and anti fungal activity of Croton roxburghii Balak. (Euphorbiaceae) stem bark
methanol extract against four pathogenic bacterial strains; two Gram positive (B.subtilis & S.aureus),
two Gram negative (E.coli and K. pneumoniae), and two fungal strains (S. flavus & C.albicans), in
different concentrations (5 μg/ml, 25 μg/ml, 50 μg/ml, 100 μg/ml, 250 μg/ml) was studied. A zone of
inhibition of extract was compared with that of different standards like Streptomycin, Cifpodoxime
and Gentamycin for antibacterial activity and Amphotericin, Fluconazole and Clotrimazole for
antifungal activity. The test drug extract showed remarkable inhibition antibacterial and antifungal
activities comparable with that of standard against the organisms tested.
KEYWORDS: Antibacterial activity, antifungal activity, Croton roxburghii stem bark, methanol
extract, Gandhamardana hills
Research article
Cite this article:
Patel Esha, Padiya RH, Acharya RN (2013), ANTIMICROBIAL EVALUATION OF CROTON
ROXBURGHII BALAK. (EUPHORBIACEAE) STEM BARK, Global J Res. Med. Plants & Indigen.
Med., Volume 2(5): 357–364
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 357–364
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
INTRODUCTION
Over the past few decades there has been
much interest in natural materials, as sources of
new antimicrobial agents. Different extracts
from traditional medicinal plants have been
tested. Many reports show the effectiveness of
traditional herbs against microorganisms and as
a result, plants have become one of the bases of
modern medicine (Evans et. al., 2002). Plants
have given the Western pharmacopoeia about
7,000 different pharmaceutically important
compounds and a number of top-selling drugs
of modern times, such as quinine, artemisinin,
shikonin and camptothecin (Tshibangu et al.,
2002). Recently much attention is being paid to
the biologically active compounds derived from
plants used in herbal medicine (Deshwal V K,
2012).
Tribal people in India used various parts of
Croton roxburghii Balak. (Euphorbiaceae)
against snake poisoning and to treat infertility,
fever and wounds (Gupta et al., 2004). In India,
only five species of Croton are used in ethno-
medicine for treatment of various diseases,
disorders and ailments like boils, bowel
complaints, chicken pox, cholera, cold and
cough, constipation, cuts and wounds,
diarrhoea, dysentery, eye diseases, epilepsy,
fever, gastric disorders, insanity, jaundice, liver
complaints, malaria, rheumatism, ringworms,
scurvy, spasmolytic agent, snake bite, sprains,
etc (Salatino et al., 2007). Its stem bark is used
as external application for sprains, bruises and
rheumatic swellings and internally in cholera,
diarrhoea, malaria, jaundice, stomach and urine
trouble, against snakebite, on inflammation etc.
(Anonymous, 2008) However no reports are
available on its antimicrobial activity except
Thatoi et Al., 2008 . Hence the study was
planned to evaluate the antimicrobial activity of
Croton roxburghii stem bark.
MATERIALS AND METHODS
Collection of plant materials
The plant Croton roxburghii Balak.
(Euphorbiaceae) was identified from its natural
habitat of Gandhamardana hill ranges,
Bolangir, Odisha, India; after studying its
morphological characters and comparing them
with the characters mentioned in various floras
(Gamble, 1993; Haines, 1988; Hooker, 1885;
Saxena, & Brahman, 1995; Shah, 1978). The
stem bark was collected, washed properly
under running water, to make them free from
foreign matter like sand, soil etc. and dried
under shade. Herbarium voucher No. 6047 was
also prepared and submitted to Pharmacognosy
museum of IPGT & RA, Jamnagar, for future
reference.
Sample preparation:
For the analysis, Croton roxburghii stem
bark was coarsely powdered to 60# and then
tests were performed.
Determination of microbial load for plant
material
Microbial load of the samples was done by
total viable aerobic count method as given in
WHO monograph (Anonymous, 1996;
Anonymous, 1998).
To 500 mg, accurately weighed sample, 1–
2 drops of Tween 80 and a homogeneous
suspension was prepared by slowly adding 5 ml
of sterile buffered sodium chloride peptone
(SBSCP) solution of pH 7.0. This suspension
was diluted 10-1
onwards as required in sterile
dilution blanks (SBSCP). One ml each from
these aliquots was added to sterile melted and
cooled top agar (Soyabean casein digest agar,
for fungal count Potato dextrose agar medium
used) tubes. These tubes were poured to sterile
petri dishes and allowed to solidify. These
plates were incubated at 30–35°C for 48 hours.
The numbers of colonies were counted and the
results were expressed in Cfu / g.
Cfu / g = Number of average colonies
Dilution × Volume plated
Antimicrobial activity of plant materials
(Anonymous, 1996)
Extract preparation: Plant extracts prepared in
methanol, 1 g of material was extracted in
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methanol by sonicating it for 10 min and then
kept overnight. Next day after filtration,
methanol was evaporated, and 5 different
concentrations i.e. 5 µg/ml, 25 µg/ml, 50
µg/ml, 100 µg/ml, 250 µg/ml were prepared
from the residue of each sample. These were
used for determination of antimicrobial
activity.
Culture conditions: The antimicrobial efficacy
of these plant materials tested on 6 different
strains, 2 Gram positive bacteria namely
Bacillus subtilis (NCIM 2063) &
Staphylococcus aureus (NCIM 2079), 2 Gram
negative bacteria namely Escherichia coli
NCIM 2065 and Klebsiella pneumoniae (NCIM
2719). Two Fungal strains namely Aspergillus
flavus (NCIM 1028) & Candida albicans
(NCIM 3471). All cultures were obtained from
NCL, Pune. 24 hour old cultures of all these
organisms were inoculated in sterile broths and
incubated till 0.5 Mcfarland standard turbidity
obtained, and then used for assay.
Antimicrobial assay (Doman & Deans, 2000)
Sterile soybean casein digest agar (25 ml/plate)
was used for antibacterial activity and sterile
sabouraud agar (25 ml per plate) was used for
antifungal activity. Medium obtained from
Himedia laboratories. 20 ml sterile medium
was poured in sterile plates aseptically and they
were kept until solidified. Then 0.5 ml of
culture is inoculated in 5 ml sterile melted and
cooled medium and poured on solidified agar
plates aseptically. After solidification well was
made with the help of cup borer and 0.3 ml of
each sample was inoculated in them. For
antibiotic discs there is no need to make wells,
disc were directly placed on agar surface
aseptically. For diffusion purpose plates were
placed in refrigerator for 20–25 minutes. Then
plates were incubated at 37°C for 24 hours
except sabouraud agar plates and plates
containing K. pneumoniae organism, they were
incubated at 30°C for 24–48 hours. After
incubation zone of inhibition was measured
with Himedia antibiotic zone scale- c.
Pathogen study (Anonymous, 1996)
Same extracts were used as for
antimicrobial activity assay. These extracts
were transferred to specialized mediums given
below and incubated at their optimum
temperature for growth, then after incubation
plates were observed and results were
concluded.
Selective differential mediums according
pathogens:
Pseudomonas aeruginosa – Citrimide agar
Salmonella typhi – TSI agar slant, XLD agar
Escherichia coli – EMB agar
Staphylococcus aureus – Mannitol salt agar
RESULTS AND DISCUSSION
Description: Croton roxburghii stem bark
powder is reddish coloured coarse powder.
Microbial load report:
The observations on the microbial load of
Croton roxburghii bark shows that the tested
samples, when collected from their natural
sources, were within prescribed limit of the
microbes. (Table 1).
Antimicrobial activity (Plate 1 & 2):
The antimicrobial activity of methanol
extracts of Croton roxburghii stem bark was
studied in different concentrations (5 μg/ml,
25 μg/ml, 50 μg/ml, 100 μg/ml, 250 μg/ml)
against four pathogenic bacterial strains two
Gram positive B. subtilis NCIM 2063 & S.
aureus NCIM 2079, two Gram negative (E. coli
NCIM 2065, K. pneumoniae NCIM 2719) and
two fungal strains (S. flavus NCIM 1028 and C.
albicans NCIM 3471). Antibacterial and
antifungal potential of extracts were assessed in
terms of zone of inhibition.
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Table 1 : Microbial load report
Parameter Sample Permissible Microbial
contamination limits
(Anonymous, 2001)
C. roxburghii stem bark
Total Viable Aerobic Count (Cfu/g)
a) Bacterial count 14.6 × 103 10
5/g
b) Fungal count 30 103/g
Pathogens (per gram)
a) S. aureus Absent Absent
b) E. coli Absent Absent
c) P. aeruginosa Absent Absent
d) S. typhi Absent Absent
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Table 2: Antibacterial activity of methanol extracts of Croton roxburghii stem bark and
standard drugs against Gram + ve and Gram -ve organisms
Sample Concentration Zone of inhibition (mm)
B. subtilis
(NCIM
2063)
S. aureus
(NCIM
2079)
E. coli
(NCIM
2065)
K.
pneumoniae
(NCIM
2719)
C. roxburghii stem bark
methanol extract
5 µg/ml 12 Nil 13 13
25 µg/ml 13 Nil 14 14
50 µg/ml 15 12 16 16
100 µg/ml 17 13 17 17
250 µg/ml 18 15 20 18
Methanol
(Control)
− Nil Nil Nil Nil
Gentamycin 10 µg 28 25 22 24
Cifpodoxime 10 µg 22 23 21 19
Streptomycin 10 µg 27 17 17 24
Figure 1: Effect of C. roxburghii stem bark methanol extract against Gram +ve and
Gram −ve strains
Figure 2: Effect of standard drug against Gram +ve and Gram −ve strains
0
5
10
15
20
25
B. subtilis (NCIM 2063)
S.aureus (NCIM 2079)
E. coli (NCIM 2065) K. pneumoniae
5
25
50
100
250
0
5
10
15
20
25
30
B. subtilis (NCIM 2063)
S.aureus (NCIM 2079)
E. coli (NCIM 2065)
K. pneumoniae
Gentamycin 10µg
Cifpodoxime 10µg
Streptomycin 10µg
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Table 3: Antifungal activity of methanol extracts of Croton roxburghii stem bark and standard
drugs
Sample Concentration Zone of inhibition (mm)
S. flavus
(NCIM 1028)
C. albicans
(NCIM 3471)
C. roxburghii
stem bark
methanol extract
5 µg/ml Nil 16
25 µg/ml 11 17
50 µg/ml 12 18
100 µg/ml 13 20
250 µg/ml 14 22
Methanol (Control) − Nil 11
Amphotericin B 50 µg 10 µg 14 19
Fluconazole 30 µg 10 µg 11 28
Clotrimazole 10 µg 10 µg 24 30
Figure 3: Effect of C. roxburghii stem bark methanol extract against two fungal strains
Figure 4: Effect of standard drugs against two fungal strains
The antibacterial and antifungal activity of
the methanol extracts of Croton roxburghii
stem bark, increased linearly with increase in
concentration of extracts (μg/ml). As compared
with standard drugs, the results revealed that in
the extracts for bacterial activity, E. coli and B.
subtilis were more sensitive when compared to
K. pneumoniae and S. aureus, and for fungal
activity C. albicans showed good results when
compared to S. flavus but S. flavus was more
0
5
10
15
20
25
S. flavus (NCIM 1028) C. albicans (NCIM 3471)
5
25
50
100
250
0
5
10
15
20
25
30
35
S. flavus (NCIM 1028) C. albicans (NCIM 3471)
Amphotericin B 50µg
Fluconazole 30µg
Clotrimazole 10 µg
Methanol (Control)
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sensitive. The growth inhibition zone measured
ranged from 12–20 mm for all the sensitive
bacteria, and ranged from11–22 mm for fungal
strains. (Table 2−3) (Fig 1−4)
The inhibitory effect of C. roxburghii stem
bark methanol extracts (5, 25, 50, 100,
250 μg/ml) showed (13, 14, 16, 17, 20 mm)
against E. coli; (13, 14, 16, 17, 18 mm) against
K. pneumonia; (00, 00, 12, 13, 15 mm) against
S. aureus; (12, 13, 15, 17, 18 mm) against B.
subtilis; (00, 11, 12, 13, 14 mm) against S.
flavus; (16, 17, 18, 20, 22 mm) against C.
albicans.
The result shows that the extracts of all
samples were found to be effective against all
the microbes tested.
CONCLUSION
In this screening work, the test drug extract
at different concentrations was found to be
effective against all organisms such as Gram
positive, Gram negative and fungal strains.
From the above results the activity of all
extracts shows significant antibacterial and
antifungal activity. The present study justified
the claimed ethnic uses of C. roxburghii stem
bark externally in ringworms, scurvy, bruises
and to treat various infectious diseases caused
by the microbes. However, further studies are
required to isolate the active compounds from
Croton roxburghii stem bark, responsible for
the antimicrobial property which may lead to
compounds in the field of antimicrobial.
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Government of India, Ministry of
Health and Family Welfare, Controller
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Pharmacopoeia of India Part-I, Volume-
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Department of Indian Systems of
Medicine & Homoepathy, New-Delhi;
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Mahendra Pal Singh, Dehradun, India,
(London); p.no. 394–395.
Salatino A, Salatino MLF, Negri G (2007).
Traditional uses, chemistry and
pharmacology of Croton species
(Euphorbiaceae). J Braz Chem Soc,18 ;
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Orissa, vol. III, Regional Research
Laboratory, Orissa Forest Development
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Orissa.(India); p.no.1323, 1372–1374.
Shah GL (1978). Flora of Gujarat State, Part-I,
Shri. K. A. Amin, Registrar, Sardar
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Thatoi HN, Panda SK, Rath SK, Dutta SK
(2008). Antimicrobial activity and
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Source of Support: Nil Conflict of Interest: None Declared
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Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
AN ESTIMATION OF HUMIC SUBSTANCES IN AN AYURVEDIC
HERBOMINERAL DRUG SHILAJATU (ASPHALTUM) AS PART OF
PHYTO-PHARMACEUTICAL STANDARDIZATION
Akarshini A M1*, Renuka
2, Shukla V J
3, Baghel M S
4
1PhD scholar, Department of Kayachikitsa, IPGT &RA, Gujarat Ayurveda University, Jamnagar, Gujarat,
India 2PhD scholar, Department of Pharmaceutical chemistry, IPGT &RA, Gujarat Ayurveda University,
Jamnagar, Gujarat, India 3Department of pharmaceutical Chemistry, IPGT &RA , Gujarat Ayurveda University, Jamnagar, Gujarat,
India 4Director, IPGT &RA, Gujarat Ayurveda University, Jamnagar, Gujarat, India
*Corresponding Author: Email: [email protected]; Mob: +919737234034
Received: 28/03/2013; Revised: 28/04/2013; Accepted: 04/05/2013
ABSTRACT
Shilajatu an important Ayurvedic drug having several medicinal properties. If administered after
proper purificatory procedure (shodhana), it has miraculous benefits. Market products are subjected
to some processing, but the purity among samples vary, thereby therapeutic efficacy too. Sothere is
need for standardization of purity assessment strategies which could help to scrutinize the good
sample. Previous efforts towards standardization of the raw drug shilajatu aimed to identify plant
source in formation of shilajatu, along with basic phyto-pharmaceutical parameters assessment.
Present work attempts to see how different the data varies when the basic phyto-pharmaceutical
parameters performed on processed sample. Also an additional assay of shilajatu, to quantify fulvic
acid through UV spectrophotometry, and quantify humic acid through gravimetric method of
analysis was carried out. Lignin decomposition logic has been attempted using UV &
chromatographic assay. Chromatography was performed as per Certified Reference Material. The
data may be used as a reference parameter for purity analysis of processed shilajatu. Results showed
the shilajatu from Nepal had high purity and greater percentage of fulvic acid compared to that from
India. This work is unique among efforts towards establishment of universal strategies in purity
assessment of shilajatu.
KEYWORDS: Shilajatu, Chromatography, UV spectrophotometry, Gravimetric method, humic
acid, fulvic acid.
Research article
Cite this article:
Akarshini A M, Renuka, Shukla VJ, Baghel MS (2013), AN ESTIMATION OF HUMIC SUBSTANCES IN AN
AYURVEDIC HERBOMINERAL DRUG SHILAJATU (ASPHALTUM) AS PART OF PHYTO
PHARMACEUTICAL STANDARDIZATION, Global J Res. Med. Plants & Indigen. Med., Volume 2(5): 365–373
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 365–373
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
INTRODUCTION
Shilajatu (Mineral Pitch) is a herbo-mineral
drug formed out from fissures of iron rich rocks
during hot weather. Traditional uses of
Shilajatu indicate its efficacy in treating
diabetes and diseases of the urinary tract as
well as edema, tumors, diseases causing
emaciation, epilepsy and insanity. Modern
science extends its indications to all the
systems of the human body with a significant
number of additions in the reproductive and
nervous system (Robert, 2004). Lots of
controversy exists, regarding the sources and
availability of pure form of shilajatu. This
article aims to analyze the difference between
samples collected from Nepal and India in
comparison to synthetic fulvic acid. This work
is unique among efforts towards establishment
of universal strategies in purity assessment of
shilajatu.
Shilajatu in its raw form contains free
radicals and may also contain mycotoxins and
fungal toxins. The processing needs to remove
the free radicals, Polymeric Quinone radicals,
toxins, mycotoxins and inactive ingredients
(Shilajit, Ayurwiki.info). Only the purified
extract gives the desired benefits of shilajatu.
So for the present work, shilajatu which had
undergone processing with triphala qwatha and
cow’s milk following the method described in
Ayurvedic Texts was obtained (Shastri Ambika
datta, 2010). Samples collected were subjected
to phyto-pharmaceutical assay comprising
quantitative and qualitative estimation of
various constituents present therein, and the
obtained results were compared. Previous
efforts towards standardization of the raw drug
shilajatu aimed to identify plant source in the
formation of shilajatu, along with basic phyto-
pharmaceutical parameters assessment
(Saileshnath saxena, 1995). Present work
attempts to see how different the data varies
when the basic phyto-pharmaceutical
parameters were performed on processed
sample. The UV spectro-photometry and
Gravimetric Methods were specially designed
for the qualitative analysis of shilajatu. Method
used for estimation of maturity of organic
compost served as pedestal for application of
above two methods here; as shilajatu itself is a
complex mixture of organic humic substance.
MATERIALS AND METHODS
Phytochemical analysis
Powder microscopy
1. Sudda shilajatu of punjab, obtained from the
pharmacy of IPGT & RA, Jamnagar, India
(Sample I).
2. Sudda shilajatu from Singh Darbar
Dawakhana, Kathmandu, Nepal (Sample II).
Shilajatu found in Nepal is considered as the
best and is also acclaimed for a highly potential
medicinal herbo-mineral drug.
Phytochemical assay of Shilajatu
Shilajatu Sample I & II were analyzed by
using qualitative and quantitative parameters at
Pharmaceutical chemistry laboratory of IPGT
& RA, Gujarat Ayurveda University, Jamnagar.
RESULTS AND DISCUSSION
Organoleptic Parameters
The characters of the sample are tabulated
in Table 1.
Microscopy of Shilajatu (Sample I and II)
Samples were subjected to microscopic
analysis after staining the samples with and
without water. The Microscopic features are
tabulated in Table 2. Images of powder
microscopy of sample I & II are shown in
Figure 1, 2.
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TABLE 1. Organoleptic parameteres of Shilajatu sample I & II
Sl.No Parameters Suddha Shilajatu Sample I Suddha Shilajatu
Sample II
1 Colour Brownish black Black
2 Odour Smell of cows urine Smell of cows urine
3 Taste Bitter, Astringent Bitter, Astringent
4 Consistency Solid Thick semisolid
5 Solubility in
Water
Easily dissolves Easily soluble
In acid
(HCl)
Dissolves in HCl with blackish brown color
changing to light brown
Slightly soluble
Alcohol Slightly soluble Insoluble
Acetic acid Slightly soluble Soluble
TABLE 2. Microscopic study of samples
Reagents used Sample I Sample II
Water High amount of crystalline material Calcium oxalate crystals
Oil globules
Crystalline material
HCl - No crystalline material
Oil globules present
FIGURE 1-Powder microscopy of Suddha Shilajatu sample I
Crystalline material Prismatic crystal
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 365–373
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FIGURE 2- Microscopy of Suddha Shilajatu sample II
Crystals in group Oil globule Prismatic crystal
Physico - chemical parameters
Sample I & II were evaluated for physico-
chemical parameters like water and methanol
soluble extractive and pH. The water and
methanol soluble extractive in sample II
measured 61.03% and 26.31% respectively and
that of sample I measured 25.2% and 18.2%
respectively. The pH was 5 in both, acidic by
nature.
Qualitative tests
The water extract of the samples were
analyzed for different functional groups.
Results of qualitative tests are shown in Table
3.
High Performance Thin layer
chromatographic study (HPTLC)
Methonolic extract of Sample I & II was
fixed through TLC. Findings of TLC shown in
Figure 3 Later HPTLC was carried out.
Methanolic extract of Sample I & II and Fulvic
acid (standard) were spotted on pre-coated
silica gel GF 60254 aluminium base plate by
Camag Linomate V sample applicator fitted
with a 100 L Hamilton syringe, 10 ml of 25%
ammonium Hydroxide : n-propanol (7:3v/v)
was used as a mobile phase. The development
distance was 6.4 cm (development time
30 min). After development, densitometric
scanning was performed with a camag TLC
scanner III in reflectance absorbance mode at
254 nm and 366 nm under control of win
CATS software (V 1.2.1 camag) (Figure 4).
The slit dimensions were 6 mm × 0.45 mm and
the scanning speed was 20 mm s-1
. Visual
observation under densitometer showed
2 spots. However the chromatogram showed
two prominent spots at hRf at 0 and 80 in
sample I and one spot at hRf 80 in sample II,
one prominent spot at hRf 80 in Fulvic acid
(standard), in short wave UV 254 nm. Result
implies both the samples contained fulvic acid.
Sample II showed high purity level whereas
sample I had constituents other than fulvic acid
too which impede its purity.
TABLE 3. Qualitative parameters for SRC for different functional groups
Sl.No Functional groups Name of the test/reagent Results
Sample I
Results
Sample II
1 Alkaloids Wagner’s Reagent Negative Negative
2 Tannin Lead acetate Positive Positive
3 Saponin Glycosides Foam test Negative Negative
4 Protein Biuret test Positive Positive
5 Steroids Libermann Burchard’s test Negative Negative
6 Flavonoids NaOH test Positive Positive
7 Reducing sugars Fehling’s test Absent Absent
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FIGURE 3. TLC of methanol extract of sample I & II
A - 260nm
B - 472nm
C - After spraying with anisaldehyde Sulphuric acid
FIGURE 4. Densitometer curve of methanolic extract of sample I, standard fulvic acid and
sample II at 254nm
UV spectrophotometry
There is no any accepted method to
quantify fulvic acid, which is practical and cost
effective. Hence it was aimed to evaluate and
develop a spectro-photometric fulvic acid
quantification protocol for future laboratory
analysis of shilajatu & other humic substances.
UV spectrophotometry was carried out to
evaluate the quantity of fulvic acid present in
the Sample I & II compared to the reference
standard fulvic acid.
Specificity to undertake UV
spectrophotometry:
Humic substances are formed by the
microbial degradation of dead plant matter,
such as lignin. These can be divided into three
main fractions: humic acids, fulvic acids, and
humin (Humate fertilizer) (Shilajatu has been
found to consist of a complex mixture of
organic humic substances and plant and
microbial metabolites come about in the rock
rhizospheres of its natural habitat. The active
principle of shilajatu is fulvic acid and it seems
to have that unique capacity to dilate and
permeate the thick cell walls to transmit the
minerals into the cells, thereby overcome
tiredness, lethargy, and chronic fatigue
(Rudramani Shilajatu). Moreover shilajatu is
not only about Fulvic acid. It contains more
than 85 minerals and nutrients which can be
instantly transported to the cells by fulvic acid
(Rudramani Shilajatu). Based on the ratio of
fulvic acid with humic acid, purity and action
potential of shilajatu can be ascertained.
Aimed to establish a quantitative
correlation between UV/viz absorption and the
concentrations of fulvic acid isolated from
different sources. Based on the assumption that
the overall optical properties of humic
substances are very similar regardless of their
C B A
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origins, this was undertaken. (Daqing Gan,
2007) An index was generated to quantify
fulvic acid in a sample with both humic acid
(HA) and fulvic acid (FA). If FA is more than
HA, then the sample is good; or vice versa. The
reactive moiety is equal to FA. This was based
on the principle that, in agricultural science
composting is believed to yield humus like
substances through various biochemical
process. This influences the quality of soil
organic matter (Carmen, 2004). Shilajatu is
also a complex form of organic humic
substances. As humic substances are formed by
the microbial degradation of dead plant matter,
such as lignin. Conversion of lignin and other
components into humic substances like humic
acids, fulvic acids, and humin is known by
Humification Index (HI). HI is used as process
controlled parameter, while in shilajatu the
humification process will be completed
naturally (not induced). Hence HI will predict
the quality of shilajatu. Thus parameter is used
to determine the quality of naturally collected
sample rather than only measuring selected
FA/HA from decomposed mass. Knowing the
ratios of HA to FA helps us predict HI, because
FAs (more aliphatic and richer in carboxiylic
acid, phenolic and quinone groups) are more
soluble and reactive than HAs (more aromatic
and insoluble when carboxylate groups are
protonated at low pH). (Carmen, 2004)
Quinones in FA have stronger antioxidant
capacity and contain higher levels of phenolics
than others which have potential antioxidant
and chemo-protective agents. (Hua-Bin- li et
al., 2011) This action is needed for
neurodegenerative diseases like DPN.
The alkaline extract of sample I & II was
prepared. After suitable dilution of one gram of
sample with 50ml of 0.5 Normal NaOH
(sodium hydroxide), it was kept overnight.
Next day the supernatant was collected, and
was scanned through 200–800 nm in a
Shimadzu UV-visible double beam recording
spectrophotometer (UV-160A) and the
absorbance in spectra were recorded based on
reference to standard range. Three standard
wavelengths 260 nm, 280 nm, 472 nm were
selected (Radosław Zbytniewski, 2005). The
UV-visible spectrums of the alkaline extract of
both samples were recorded Results observed
under UV–visible spectrum has been presented
in Table 4. It shows two absorption peaks at
472 nm and 664 nm, negative peaks at 280 nm,
260 nm. There is no absorption in the visible
region. The following absorbance ratios was
used for calculation (Kononova, M., 1968,
Gieguzy_nska E, 1998)-
Q2/6(HA) = Absorbance Ratio of 260/664,
denotes the relation between non-humified and
strongly humified material. No decay started.
Q4/6(FA) = Absorbance Ratio 472/664, is
often called the humification index. Typical
values of the Q4/6 ratio for humified material
are usually <5. (Gieguzy_nska E, 1998)
Q2/4(lignin & others) = Absorbance Ratio
280/472, reflects the proportion between the
lignins and other materials at the beginning of
humification, and the content of materials at the
beginning of transformation.
Low Q2/6 or Q4/6 ratios reflect a high degree
of aromatic condensation and indicate a higher
level of organic material humification.
(Carmen, 2004)
As per index sample II contained good
amount of FA (3.823) in comparison to sample
I (3.071). The standard sample showed only
1.666 of fulvic acid. Table 4.
The UV spectrum is much practical, rapid
means of estimating the concentration of a
fulvic acid solution. It estimates characteristic
pH independent, exponential spectrum of a
standard FA in the 200 – 800 nm wavelength
range. Given the length and complexity of FA
sample isolation and the likelihood of at least
some sample loss on the chromatography
columns involved, this simple spectro-
photometric approach described here (Elham A,
2009) merits application for the analysis of
fulvic acid solutions. The results of this work
have potential use for the routine analysis of
FA samples and for the certification and
regulation of commercial shilajatu and its
formulations.
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 365–373
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TABLE 4 - Qualitative parameters for SRC for different functional groups
Ratio FA Sample I Sample II
Q1= HA −10.703 −16.357 −9.000
Q2= FA 1.666 3.071 3.823
Q3=lignin and others −5.288 −6.0697 −1.692
TABLE 5 - Showing the absorbance at different wavelengths
Sample 280 nm 472 nm 664 nm 260 nm
Shilajatu I −0.261 0.043 0.014 −0.229
Shilajatu II −0.220 0.130 0.034 −0.306
Fulvic acid −0.238 0.045 0.027 −0.289
Gravimetric method/Acid precipitation
method
The acid precipitation method has been
widely accepted for the separation and
subsequent quantification of humic acid
(Thurman EM, 1981). The quantification of
humic substances is important because humic
materials have a relatively high content of free
radicals which play important roles in
polymerization and redox reactions. This
affects the mobility of metals (both those with
nutrient value and those that are of concern
because they are pollutants (Carmen, 2004).The
humic acid (HA) precipitates at pH < 2 and
thus can be quantified by gravimetric
measurements. A comparison of appropriate
methods of HA analysis favors HA
precipitation from alkaline solution by addition
of concentrated HCl followed by washing of
the precipitate with water and oven drying at
110oC (Fataftah AK, 2001). The humic and
fulvic acids were extracted into a strongly basic
aqueous solution of sodium hydroxide. This
was made acidic by adding normal
hydrochloric acid (6N HCl) by adjusting the pH
up to 2 and kept overnight. Precipitate obtained
was filtered and was oven dried at 110 o C. HA
is precipitated from this solution, leaving the
fulvic acids in solution. This measure implies
to the weight of HA in sample I & II. This
measure should be low to say if good sample.
This is the operational distinction between
humic and fulvic acids HA in sample II (-
9.000) was less compared to sample I(-
16.357).The standard showed −10.703 of HA.
Table 5.
By this procedure, Sample I contained
0.320 gms, sample II 0.546 gms and standard
contained 0.094gms of humic acid out of 1 gm
of samples. This high amount of humic acid
compared to standard may be due to the
method followed for purification of sample I &
II. Subjection of drugs to processing might
have switched humification. Decomposition is
controlled by climatic condition (Choppin
1985), induced naturally. So based on the level
of humification the quantity of humic acid
could be varied.
CONCLUSION
Humic substances are used in medical
science with substantial benefits in improving
the health status. Shilajatu is one such complex
mixture of humic substance used widely for its
strengthening and rejuvenative qualities.
Market products are subjected to some
processing, but the purity among samples vary,
thereby therapeutic efficacy too. So there is
need for standardization of purity assessment
strategies, which could help to scrutinize the
good sample. There are no universal standards
established to quantify the humic substances in
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 365–373
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
shilajatu. Knowing the ratio of FA and HA
helps to predict the activity potential of
shilajatu and its purity. Keeping this in mind
the UV spectrophotometer and Gravimetric
methods were specially designed for easy and
cost effective qualitative analysis of shilajatu.
Method used for estimation of maturity of
organic compost served as pedestal for
application of above two methods here. It is
reported that both the samples meets the
minimum standards as reported in API at the
preliminary level. The shilajatu sample
collected from Nepal showed better level of
fulvic acid comparatively. The results of this
work have potential for the routine analysis of
FA samples and for the certification and
regulation of commercial shilajatu and other
fulvic acid containing drugs. Thus inference
from this study may be used as reference
method in further quality control researches.
ACKNOWLEDGEMENTS
Harisha CR, Head of Pharmacognosy
Laboratory, for carrying out pharmacognostical
works related to work and Prof. Prajapati PK,
Director of Pharmacy, Department of
Rasashastra, IPGT & RA, Gujarat Ayurveda
University, Jamnagar for his guidance during
the work.
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Hindi commentory(2010), Shilajatu
shodhana prakarana. Chowkamba
Amarabharati Publications, Varanasi:
chap 2/111 pp. 53
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Source of Support: Nil Conflict of Interest: None Declared
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Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
MANAGEMENT OF ARBUDA (CANCER) WITH HERBOMINERAL
FORMULATION - A PILOT STUDY
Mahanta Vyasadeva1*, Dudhamal T S
2, Gupta S K
3
1,2 Assistant Professor, Shalya - Tantra, IPGT & RA. Gujarat Ayurved University, Jamnagar.
Gujarat, India 3
Associate Professor & I/C HOD, Shalya - Tantra, IPGT & RA. Gujarat Ayurved University,
Jamnagar, Gujarat, India
*Corresponding Author: Email: [email protected]; Mobile: +91 9408323779
Received: 26/03/2013; Revised: 14/04/2013; Accepted: 25/04/2013
ABSTRACT
Arbudaharana rasayana is an anubhuta yoga (Self experienced compound medicine). It is a
herbo-mineral compound formulation, practicing by traditional healers of Orissa as a folklore
medicine, for the management of Arbuda. The formulation was studied clinically in 10 patients of
non operable malignancy of different regions of body like scalp, vagina, oesophagus, bladder and
oral cavity. The drug was given in powder form, 5 gm twice a day, orally with Luke warm water for
two months. Symptomatic relief was observed in treated patients with overall improvement in quality
of life. The drugs might have been effective due to anti-cancerous, antioxidant and immuno-
modulator activities. During follow up period there was no any adverse effect of drug observed.
KEYWORDS: Anti-cancerous, Anti-oxidant, Arbuda, Arbudaharana rasayana, Immuno-modulator
Research article
Cite this article:
Mahanta Vyasadeva, Dudhamal T S, Gupta S K (2013), MANAGEMENT OF ARBUDA (CANCER)
WITH HERBOMINERAL FORMULATION - A PILOT STUDY, Global J Res. Med. Plants &
Indigen. Med., Volume 2(5): 374–379
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 374–379
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
INTRODUCTION:
Globally, the burden of new cancer cases is increasing day by day even after several advances made in treatment field of cancer. As per the WHO data, about 70% of all cancer death occurs in low and middle income country (http://www.who.int). Worldwide death from cancer is projected to continue to raise over 11 million in 2030. Every year in India, about 11 lakh new cases are registered and approximately 5 lakh patients die due to cancer. In India, it has been estimated that about 71% of deaths occur due to cancer belongs to 30–69 years age groups of people (Dikshit R et al., 2012). As etiological factors, tobacco consumption, low fibre diet intake, lack of physical activities, eating much fast food and alcohol intake are usually linked with prevalence of the cancer directly or indirectly (http://www.cancerfoundationofindia.org, http://www.cancer.gov). In addition, viral infections of HBV/HCV and HPV are also held responsible for about 20% of cancer deaths.
To manage the excessive proliferation of cells and to control the metastasis of abnormal cells by mutation (RCG Russel et.al. 2004) is still remained as a challenge before physicians. Now a day, multi modality treatment is being practised even then the outcome and prognosis is not found satisfactory due to poor awareness and late detection of cancer. The contemporary treatment like chemotherapy, radiotherapy and surgical intervention are still not affordable by poor patients and those receiving such therapy, they also need some adjuvant therapy like immunotherapy to overcome untoward effects. At this juncture, Ayurveda can extend good support as a safe, effective and more affordable treatment to improve the overall quality of life (QOL).
As per Ayurveda, manifestation of various non inflammatory swelling occurs due to vitiation of Vata and Kapha Dosha and they are classified as Arbuda (tumour), Granthi (glandular swelling). Galaganda (cervical lymphadenopathy) etc. (Shastri A, 1995). One can correlate the Arbuda with cancer on the basis of its signs and symptoms. The definite aetiological factors of Arbuda are not defined in Ayurvedic classics but the causative factors defined for Granthi and Vranasopha have been
considered for management of Arbuda. Acharya Sushruta has given more emphasis for surgical management of Adhyarbuda (tumour grows over the pre existing one), Dwirbuda (tumour grows simultaneously or one after other) and Mamsarbuda (fleshy tumour) (Singhal G.D, 1972, Ghanekar B, 1977) to improve the quality of life. He has also advised to go for eshansi Samudharet (En-block resection) of Arbuda to prevent recurrence. Maggots’ therapy and Shodhana therapy have been advised in non operable conditions as palliative therapy.
“No disease is manifested without involvement of Tridosha” is the basic ideology of Ayurveda and one can control the vitiation of Tridosha with the help of herbal / herbomineral formulations. The formation of any Arbuda can occurs in any of dushya i.e. Rakta (blood), Mamsa (muscle) and Meda dhatu (fat) due to vitiation of Vata and Kapha dosha. The vitiated Vata and Kapha dosha produce Ama (autotoxin) by affecting the Jatharagni and Dhatwagni. That Ama along with apakwa dhatu produces various dhatugat vikara in the form of Arbuda, Shopha, Granthi etc. by obstructing the Srotasa (channels).
Hence, the involvement of Agni, Ama and Srotavarodha are to be considered during the management of Arbuda. On the basis of such concept Arbudaharana Rasayana, as a folklore medicine, is being practiced by traditional vaidyas of Orissa for the management of Arbuda (cancer). For present study, total 10 patients of non operable malignant cases of scalp, vagina, oesophagus, colon, bladder and oral cavity were selected from the OPD of Dept. of Shalya-Tantra.
MATERIALS AND METHODS:
Total 10 diagnosed cases of cancer of various parts of the body, were selected irrespective of age, sex and religion for this study. All the selected patients were having features of local and distant metastasis. Different types of cancer patients were included in this study as a pilot study.
The Arbudaharana Rasayana (anubhuta yoga) is the combination of following herbo-mineral drugs having anticancerous, antioxidant, immuno-modulator activities.
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 374–379
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Method of drug administration to the patients
To
Sr.
No.
Name of Drug Ingredients Quantity
/ Dose
1. Haridrakhanda Haridra (Curcuma longa), Ghrita (Ghee), Godughdha (Cow
milk), Sarkara (Sugar), Trikatu (Combination of Zingiber
officinale, Piper logum, Piper nigrum), Trijataka
(Combination of Cinnamomum zeylanica, Cinnamomum
tamala, Elettaria cardamomum), Vidanga (Embelia ribes),
Trivrit (Operculina turpethum), Triphala (Combination of
Terminalia chebula, Terminalia bellerica, Phyllanthus
emblica), Keshar (Crocus sativus), Musta (Cyperus rotundus),
Louha (Iron)
4 g
2. Arogyavardhini Parada (Mercury), Gandhak (Sulphur), Louha (Iron), Tamra
(Copper), Abhrak (Mica), Triphala (Combination of
Terminalia chebula, Terminalia bellerica, Emblica
officinalis), Shilajatu (Asphatum panjabinum), Guggulu
(Commiphora mukul), Chitrak moola (Plumbago zeylanica),
Katuki (Pichrorhiza kurroa)
250 mg
3. Vydadhiharan
Rasa
Parada (Mercury), Gandhak (Sulphur), Somal (White
arsenic), Haratal (Yellow arsenic), Manashila (Red arsenic) ,
Rasakarpur (Per chloride of mercury)
150 mg
4. Guduchi satwa Guduchi (Tinospora cordifolia) 600 mg
Posology – 5 g powder twice a day (In morning, after breakfast and in evening after light snacks)
orally with Luke warm water.
RESULTS:
Demographic Analysis:
Out of 10 patients only 20% female patients
were found in age group of 30–40 years
whereas above 40 years age 80% patients were
recorded suffering from different types of
cancer. The data itself revealed that the cancer
is occurred in older age (Table No.1).
In case of female patients except cervix and
vaginal cancer 20% and breast cancer 10% no
other site of cancer like head & neck,
oesophagus, lungs, colon and urinary bladder
were found as reported in cases of male
patients (70%). This data show that there are
more chances of occurrence of these sites of
cancer in male population (Table No.2).
Incidences of squamous cell carcinoma
(60%) were found more in comparison to
adenocarcinoma (30%) and fibroadenoma
(10%). (Table No.3)
Maximum addiction of tobacco, smoking
and tea (70% each) were observed in male
patients and may be held responsible more
incidences of cancer in male population (Table
No.4).
The data of socio-economical status
revealed that maximum lower middle (50%)
and lower (20%) classes of patient were
suffering from different sites of cancer which
show that cancer may have relation up to some
extent to the socio-economical status of the
people (Table No.5).
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Table No.1 Age wise distribution n-10
Sr. No. Age (Years) Male % Female % Total %
1. 30–40 00 00% 02 20% 02 20%
2. 41–50 04 40% 01 10% 05 50%
3. 51–60 02 20% 00 00% 02 20%
4. >60 01 10% 00 00% 01 10%
Table No.- 2; Analysis of Cancer Sites n- 10
Sr. No. Sites of Cancer Male % Female % Total %
1. Head and neck 02 20% 00 00% 02 20%
2. Oesophagus 01 10% 00 00% 01 10%
3. Lungs 01 10% 00 00% 01 10%
4. Colon 01 10% 00 00% 01 10%
5. Urinary Bladder 02 20% 00 00% 02 20%
6. Cervix and Vagina 00 00% 02 20% 02 20%
7. Breast 00 00% 01 10% 01 10%
Table No.- 3 Histopathological Analysis n- 10
Sr. No. Tissue / Cell type Male % Female % Total %
1. Squamous cell
carcinoma
04 40% 02 20% 06 60%
2. Adeno carcinoma 03 30% 00 00% 03 30%
3. Fibroadenoma 00 00% 01 10% 01 10%
Table No.- 4 Addiction wise Analysis n-10
Sr. No. Habits Male % Female % Total %
1. Tobacco chewing 07 70% 01 10% 08 80%
2. Smoking 07 70% 00 00% 07 70%
3. Tea 07 70% 03 30% 10 10%
4. Alcohol 02 20% 00 00% 02 20%
Table No.- 5 Analysis of Socio-economical Status n-10
Sr. No. Social status Male % Female % Total %
1. Upper 01 10% 00 00% 01 10%
2. Upper middle 01 10% 01 10% 02 20%
3. Lower middle 03 30% 02 20% 05 50%
4. Lower 02 20% 00 00% 02 20%
DISCUSSION:
Outcome is the key of success in cancer
management. As cancer is a leading cause of
death recorded in the world. An integrative
approach of treatment is the need of the time to
improve the quality of life in non operable and
non tolerable cancer cases to radiotherapy /
chemotherapy. The Arbudaharana Rasayana
(anubhuta yoga) formulation contains
ingredients like Haratal (arsenic)
(http://www.wakehealth.edu), a good source of
anti-cancerous drug which helps to check the
growth of tumour by inhibiting the mutation of
the cells. Other ingredients like Guduchi
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 374–379
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
(Tinospora cordifoloia) and Haridra (Curcuma
longa) (Ranjith M. S, 2008; Bharat B, 2003)
have antioxidant, immuno-modulator, analgesic
and anti inflammatory effects which might
have extended relief by reducing inflammation
and pain.
As it is considered that Amadosha and
Srotavorodha are the known factors for
causation of Shotha and Vedana (inflammation
and pain), the Vydadhiharan Rasa (Rasatantra
Sara, 1990)is capable to remove Srotavorodha
by digesting Amadosha with increasing
Dhatwagni (cellular metabolism) and rendered
relief in the features of Shotha and Vedana.
Due to dominance of Vata Dosha in older
age and various addiction like smoking,
tobacco chewing are the favourable factors for
developing cancer. In this study 70% patients
were older and addicted for smoking and
tobacco chewing and they were developed
cancers on different site. Socio-economical
status of people may be related with the
nutrition, immunity and overall health of a
person. The data of this study also goes in
favour of that and the causes of cancer may be
attributed to the status of nutrition and
immunity as it is obviously low in socio-
economically week patients.
Squamous cell carcinoma and
adenocarcinoma were found more in this study
which showed the involvement of the particular
organs like head & neck, oesophagus, lungs,
colon and urinary bladder, etc. It may have
some relation with food and addiction habit of
the persons. In Gujarat, tobacco in the form of
Mawa - Masala, spicy and fast foods are the
favourable items for routine consumption.
These may be held responsible for causation of
cancer by developing Amadosha and
Srotavorodha with help of vitiated Vata and
Kapha.
The overall effect of the selected
formulation in the cancer patients was found
satisfactory with no any untoward effect
observed.
CONCLUSION:
Cancer is a major and global health
problem and the outcome of any available
therapy is still under the question, particularly
in non operable cases of malignancies. The
Arbudaharana Rasayana (Anubhuta yoga) is a
potent formulation for providing symptomatic
relief from Arbuda (cancer). The formulation
was found effective to improve the quality of
life (QOL) by rendering anti inflammatory,
analgesic, anti oxidant and immuno-modulator
activities.
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Source of Support: Nil Conflict of Interest: None Declared
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ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
IMPORTANCE OF UPAYOGASAMSTHA (DIETETIC RULES) IN RELATION
TO DIGESTION OF THE FOOD
Avhad Anil D1, Vyas H A
2, Dwivedi R R
3
1Ph.D. Scholar, Department of Basic Principles, Institute for Post Graduate Teaching & Research in
Ayurveda, Gujarat Ayurved University, Jamnagar - 361 008, Gujarat, India. 2Assi. Professor, Department of Basic Principles, Institute for Post Graduate Teaching & Research in
Ayurveda, Gujarat Ayurved University, Jamnagar - 361 008, Gujarat, India. 3Professor& HOD, Department of Basic Principles, Institute for Post Graduate Teaching & Research in
Ayurveda, Gujarat Ayurved University, Jamnagar - 361 008, Gujarat, India.
*Corresponding Author: E-mail: [email protected]
Received: 14/03/2013; Revised: 17/04/2013; Accepted: 20/04/2013
ABSTRACT According to Ayurveda food is one of the important factor for health as well as source for
diseases i.e. wholesome and unwholesome food is responsible for happiness and misery
respectively.. Beside the quality and quantity of the food it is very much important that how it is
taken, because food taken in improper way can lead to various diseases. Keeping these things in
mind 8 factors (Aharavidhivisheshayatana) has been mentioned in Vimanasthana of Charak Samhita
which determines the utility of the wholesome food. ‘Upayogasamstha’ is the eighth factor among
these which deals with the dietetic rules. Further in the sixth chapter of Sharirasthana six factors
which are necessary for the transformation and digestion of the food (Aharaparinamakara bhava)
have been mentioned. The study was planned to evaluate the relation between ‘Upayogasamstha’
(dietetic rules) and digestion of the food. It is found that each and every rule mentioned in
Upayogasamstha (dietetic rules) has very essential role in the process of digestion. Conflictions of
these rules are directly mentioned as the causative factors for the production of Ama (undigested
food), which is the cause for several diseases. Along with balanced diet, incorporating the dietetic
rules in our daily routine can prevent many diseases. All the rules mentioned under
‘Upayogasamstha’ (dietetic rules) strengthen the Aharaparinamakar Bhavas (digestive factors) and
also governs the digestion process along with Agni.
KEY WORDS: Aharavidhivisheshayatana, Upayogasamstha, Aharaparinamakara bhava, Dietetic
rules
Review article
Cite this article:
Avhad Anil D, Vyas H A, Dwivedi R R (2013), IMPORTANCE OF UPAYOGASAMSTHA (DIETETIC
RULES) IN RELATION TO DIGESTION OF THE FOOD, Global J Res. Med. Plants & Indigen. Med.,
Volume 2(5): 380–385
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 380–385
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
INTRODUCTION
Ayurveda being the science of life deals
with several aspect of the human life and also
cares a lot about the factors responsible for its
maintenance and prosperity. Ahara (Food),
Nidra (Sleep) and Bramhacharya (Celibacy)
are said to be the tripods of human life and
proper balance of them is responsible for the
health of an individual (Tripathi Ravidutt,
2005). Among these also ‘Ahara’ is considered
as the chief factor since the complexion,
longevity, happiness, satisfaction, nourishment,
strength and intellect are all conditioned by it
(Tripathi Ravidutt, 2005). In the conceptual
part it has been found that all the authors of
Ayurveda have emphasized on Ahara (Food) as
well as its Matra (quantity), Kala (Time of
preparation), Samskar (processes of
preparation), Desha (place; including soil,
climate and surrounding conditions), Swabhava
(constitution) and Ashanavidhi (codes and
conducts of taking food). Modern science has
particularly described Ahara according to the
nutritional value of its components. They have
not described about Hita (wholesome), Ahita
(unwholesome), Pathya (healthy), Apathya
(unhealthy) Ahara etc. for each individual.
There is no constraint of diet imposed in the
Allopathic methods of treatment, where
medicines are given principal importance. But
it is not so in the case of Ayurveda where, on
the contrary, appropriately selected and
controlled diet is an integral part of treatment
so as to enhance vitality and regulate the
medicinal effects in natural harmony with the
entire system of the body and mind .
For the sustenance of an individual
digestion plays an important role. The good
quality food taken in appropriate amount will
nourish the tissue elements of the body along
with mind and senses only when it is properly
digested. ‘Agni’ is considered as the key factor
in the process of digestion and metabolism, and
all the diseases are said to be produced due to
the weakness of Agni (Atridev Gupt, 1997).
Along with ‘Agni’ there are many co-
supportive factors which also control the
digestive process as the same type of food
could have contrary effects, under different
circumstances or for different persons; most
important is what guidelines one follows in the
selection and preparation of food and in his
eating habits. In today’s fast paced life, many
norms regarding diet & regimen have been
compromised which is showing the overall
declining status of health in the society. Ahara
and its related codes and conducts have been
long forgotten by the human being; hence to
evaluate this ancient science of dietetics this
study was undertaken.
The present review deals with
‘Upayogasamstha’ (dietetic rules) which
governs the process of digestion as all these
rules can be included under the factors which
are necessary for the digestion. Also this
review explores the importance of
Upayogasamstha (dietetic rules) and evaluates
the relation between Upayogasamstha (dietetic
rules) and digestion.
MATERIALS
The data from various texts (Tripathi Ravidutt,
2005) along with Brihatrayee has been
collected, compiled and analyzed for the
discussion and attempt has been made to draw
some conclusions.
DISCUSSION
Ayurvedic nutrition is a vast topic that takes
into account the individual constitution, the
medicinal value of culinary spices, the theory
of shad rasa (or six tastes, which should all be
present for a meal to be balanced), and more.
The founders of Ayurveda had affirmed food as
the essential source of sustenance and
strengthening of health. However, they had also
alerted that "what one eats and how?" could be
the major factor for or cause of illness.
According to modern science, ‘people who are
eating according to the rules of a traditional
food culture are generally healthier than those
of eating a modern western diet of processed
foods.
Acharya Charaka has described ‘Ashta
Aharavidhivisheshayatana’ (eight factors
determining utility of the food) (Tripathi
Ravidutt, 2005). These eight factors are
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associated specifically with the useful and
harmful effects of the food and they are
conditioned by one another. ‘Upayogasamstha’
is the eighth factor among these which
indicates the dietetic rules. Healthy as well as
diseased individuals should follow these rules
even while using wholesome food by nature.
According to Charaka; Ushma (heat), Vayu,
Kleda (moist), Sneha (Unctuousness), Kala
(time) and Samayoga (appropriate intake) are
the ‘Aharaparinamakara bhavas’ (Digestive
factors) (Tripathi Ravidutt, 2005). The rules
mentioned under Upayogasamstha and their
relations with Aharaparinamakara bhava are as
follows
i. Ushnam Bhunjit (Food should be warm)
When the food is taken warm, it becomes
appetizing and after intake it provokes the
factors responsible for the digestion. It gets
digested quickly and helps in the downward
movement of Vata and alleviation of Kapha.
This warmness indicates the ‘Ushma’ (heat),
which is the most essential and vital factor
responsible for the digestion as it directly takes
part in the digestion (Tripathi Ravidutt, 2005).
On the other hand, when the food is kept for
longer time after preparation or when stale food
is consumed, it takes more time to digest or
sometimes may produce Ama (undigested food)
(Tripathi Ravidutt, 2005). In modern science
also it has been mentioned that stale food can
lead to food poisoning at times.
ii. Snigdham Bhunjit (Food should be
unctuous)
The unctuous food is said to be delicious
and after intake it provokes the subdued power
of digestion. It gets digested quickly, helps in
Vatanulomana (downward movement of Vata),
promotes growth of the body (Sharira
upachaya), strengthens the sense faculties, and
brings out the complexion.
It indicates the third and fourth factor of
digestion i.e. Sneha (unctuousness) and Kleda
(moist). Due to them the food particles become
soft and well dissolved, so that they can be
easily digested. On the contrary Ruksha ahara
(food having dry nature) is mentioned as one of
the causative factor for the production of Ama
(Tripathi Ravidutt, 2005) and also can lead to
Vatavyadhis (disorders caused by Vata dosha).
iii. Matravat Bhunjit (Food in proper
quantity)
When the food is taken in proper quantity,
it promotes the longevity of life without
afflicting Vata, Pitta and Kapha. It does not
impair the power of digestion and gets digested
without any difficulty. For the proper digestion
one part of the stomach should be left for the
free movement of Vata, Pitta and Kapha. In
modern science also it has been said that,
always leave the table a little hungry; many
cultures have rules that you stop eating before
you are full. In Japan, they say eat until you are
four-fifths full. Islamic culture has a similar
rule and in German culture they say, ‘tie off the
sack before it’s full’ (Michael Pollen, 2006).
Hence this rule can be correlated with the
second factor necessary for digestion i.e. Vayu.
Prana Vayu (one among the 5 subtypes of Vata
dosha) takes the food nearer to the site of Agni
and Samana Vayu (one among the 5 subtypes
of Vata dosha) stimulates Agni, facilitating the
process of digestion (Tripathi Ravidutt, 2005).
If one fills his stomach with excess of the food
then it leads to obstruction in free flow of the
‘tridoshas’ due to which the digestion becomes
uneasy. Atimatrashana (Excessive intake of
food) is mentioned as the major factor for the
production of Ama (Tripathi Ravidutt, 2005)
whereas Pramitashana (less intake of food) is
described to produce emaciation of the body.
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 380–385
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Table 1: Relation between Upayogasamstha and digestion
Sr. no. Upayogasamstha Digestive factors
1 Ushnam Ushma
2 Snigdham Sneha, Kleda
3 Matravat Vayu
4 Jeerne, Natidrutam, Nativilambitam Kala
5 Veerya aviruddha, Ishta deshe Ishta sarvopakarane,
Tanmana bhunjit, Atmanamabhisamikshya
Samayoga
iv. Jeerne ashniyat (Intake after digestion of
previous meal)
According to Ayurveda, one should take the
food after the complete digestion of previous
food. Such food promotes the longevity of the
body and keeps the Doshas in balanced state.
On the other hand, when an individual takes
food before the digestion of previously taken
meal, then it gets mixed with Apakva
Ahararasa (partially digested food) of previous
meal; which leads to the vitiation of all the
three Doshas instantaneously.
This rule is nothing but the fifth factor of
the digestion process i.e. Kala (time). Even in
the presence of all other factors, digestion
requires time for completion of the process.
Hence Kala (time) is described as one of the
major factor as it brings the maturity of the
process of digestion. Vishamashana (intake of
food at wrong time), Adhyashana (intake of
excess food), Ajirnashana (intake of food
before the digestion of previous food) etc.
known causative factors can be included under
the title of Akalabhojan (intake of food at
wrong time) which is nothing but opposite of
Kalabhojana (intake of food at proper time).
v. Veerya aviruddha ashniyat (Intake of food
having no contradictory potencies)
One should take the food having no
contradictory potencies. By taking such food
one does not get afflicted with diseases and all
the attributes of the food are transformed into
the tissue elements. If the properties of food
articles are contradictory against each other,
one may suffer from the diseases like Kushtha
(skin disease), Visarpa (cellulitis) etc (Tripathi
Ravidutt, 2005), and also such food ruins the
body gradually (Tripathi Ravidutt, 2005).
vi. Ishta deshe, Ishta sarvopakarane (Intake
in proper place and with all accessories)
Desha is such a factor which is to be
understood in three ways in relation to food i.e.
Bhoomi (soil), Ahara Dravya Desha
(originating place of the food material) and
Atura Desha (eating place). Here in this context
it is mainly related with the latter one.
According to Ayurveda an individual should
take the food at proper place which is clean and
desired. Food consumed at unhygienic or
undesired place may disturb the mindset
leading to indigestion. In modern science also it
has been revealed that hundreds of millions of
people are affected by preventable diseases
originate in the environment in which they live.
Further one should be equipped with all the
necessary equipments while taking the food;
because in absence of these, an individual may
not be satisfied even with the good quality of
food.
vii. Natidrutam (Intake not in hurry)
One should not take food too speedily. If
food is taken too hurriedly it enters into a
wrong passage; it gets depressed and it does not
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 380–385
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
enter into the stomach properly. In this
situation one can never determine the taste of
food articles and will lead to improper mixing
of Bodhaka Kapha (one among the 5 subtypes
of Kapha dosha), which softens the food
particles and thereby facilitates the process of
digestion. Further, an individual taking food
speedily is unable to detect the foreign bodies
like hair etc., mixed with food.
viii. Nativilambitam (Intake not too slow)
One should not take food very slowly
because this will not give satisfaction to the
individual. In this situation he would take more
than what is required; the food would become
cold and there will be irregularity in the
digestion process. Also by taking food very
slowly, enzymes responsible for digestion will
come late in contact with food and in an
irregular manner and this will result in irregular
digestion.
The above mentioned two rules (seventh
and eighth) are also related with the factor Kala
(time). Hence they again emphasize the
importance of Kala (time) in digestion process.
ix. Tanmana Bhunjit (Intake with
concentration)
One should not talk or laugh or be
unmindful while taking food. An individual
taking food while talking, laughing or with
detracted mind, subjects himself to the same
trouble as one eating too hurriedly. In modern
dietetics also it is said that, ‘If we eat while we
are working or while watching TV or driving,
we eat mindlessly and as a result eat a lot more
than we would’ (Michael Pollen, 2006).
x. Atmanamabhisamikshya (Intake with self
confidence)
One should take food in a prescribed
manner; with due regard to his own self. The
knowledge of the usefulness of food articles is
the main thing for self-preservation.
Rule number five, six, nine and ten can be
understood under the broad heading of
‘Samayoga’ (appropriate intake) which is the
last factor necessary for digestion. Samayoga
means appropriate intake of the food which
brings the equilibrium of Dhatus. In absence of
this even the wholesome food is not digested
well.
CONCLUSION
In Ayurveda food plays a prominent role in
promoting health and is therefore considered
medicine. Ayurvedic concept of digestion takes
into account many things viz. food, its quantity,
place, time, factors affecting it, dietetic rules,
digestive factors, Agni etc.; which is the most
ideal than any other science. Along with Agni
there are many other factors which are found
useful for proper digestion and by taking care
of which one can eliminate the causes of
imbalance. Each and every rule mentioned in
Upayogasamstha has very essential role in the
perspective of digestion. Conflictions of these
rules are directly mentioned as the causative
factors for the production of Ama, which is the
cause for several diseases. Along with
balanced diet, incorporating the dietetic rules in
our daily routine can prevent many diseases.
All the rules mentioned under
‘Upayogasamstha’ (dietetic rules) strengthen
the ‘Aharaparinamakar Bhavas’ (digestive
factors). Hence these ten rules are directly
related with the ‘Aharaparinamakar Bhavas’
(digestive factors) and govern the digestion
process.
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REFERENCES
Atridev Gupt (1997), Ashtang Hriday with
Vidyotini hindi commentary by, twelth
edition, Chaukhamba Sanskrit
Bhawan, Varanasi, Nidanasthana 12.
Michael Pollen (2006), The Omnivore’s
dilemma: a natural history of four
meals, second edition, Penguin press,
New York.
Tripathi Ravidutt, (2005), Charaka Samhita
edited with Vaidyamanorama Hindi
commentary by Reprint edition,
Chaukhamba Sanskrit Pratisthana,
Delhi, Sutrasthana 11, 25, 26;
Vimanasthana 1, 2; Sharirasthana 6;
Chikitsasthana 15.
Source of Support: Nil Conflict of Interest: None Declared
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 386–391
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
CONCEPT OF VYADHIKSHAMATVA (IMMUNITY) AND ITS
RELATIONSHIP WITH BALA (VITAL STRENGTH)
Sharma Mahesh Kumar1*
1Assistant Professor, Department of Basic Principles, MSM Institute of Ayurveda, BPS Mahila
Vishwavidyalaya, Khanpur kalan, Haryana (131305)
*Corresponding Author: E-mail: [email protected]; Mob: +919468342229;
Received: 26/03/2013; Revised: 17/04/2013; Accepted: 20/04/2013
ABSTRACT
When a group of people exposed to a particular disease, the affect seen is categorized in four
categories mild, moderate, severe and unaffected. The pathogenic factors require some essential
favorable conditions to flourish and create the disease. Ayurveda finds Bala and Vyadhikshamatava
as profound explanations for this phenomenon. These are also necessary for prevention and rapid
recovery from illness. Bala is the cause for the good defense mechanism and to carry out all the
physical and psychological actions. The depreciation of bala is a constant sign observed in various
degenerative diseases and recurrent infections. It can be increased by intake of wholesome food,
immunomodulator drugs like Tinospora Cordifolia Willd. (family- Menispermaceae) etc., a regular
routine of seasonal and daily regimens along with mild physical workout. So, bala and
vyadhikshamatva have cause and effect relationship.
KEY WORDS: bala, immunity, vyadhikshamatva.
Review article
Cite this article:
Sharma Mahesh Kumar (2013), CONCEPT OF VYADHIKSHAMATVA (IMMUNITY) AND ITS
RELATIONSHIP WITH BALA (VITAL STRENGTH), Global J Res. Med. Plants & Indigen. Med.,
Volume 2(5): 386–391
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 386–391
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
INTRODUCTION
In our daily experiences with the
continuously changing environment (Shastri
Ambikadutta, 2003) we came across many
infecting agents. The external environment also
checks the adaptive power of humans (Trikamji
Yadavaji, 2005). Kala (time) causes the caya
(deposition), parakopa (vitation) and
prasamana (normalization) of the doshas to
affect the body and the parakopa make one to
suffer with the diseases. The nature has made
the natural safety measures to claim superiority
over such natural influences that hamper life.
The nature had given us power to adapt and
overcome these variations. This adaptability of
the body is termed as vyadhikshamatva
(Trikamji Yadavaji, 2005). In terms of medical
science it is called immunity. It is the diversity
of life that life exists in enormous forms
starting from the smallest viruses and bacteria
up to the largest one. All these creatures are the
miracles of nature that somehow affect each
other by causing them diseases and take their
life too. The body with adequate
vyadhikshamatva is capable to overcome the
effect of these pathogens to make oneself
disease free or with the mild prevalence. The
one who possess good vyadhikshmatva has a
good dehabala (Physical health) (Jangid
Chhagan et.al 2008). The good physical health
means healthy body metabolism etc. Hence this
review explores the relationship between
Vyadhikshmatva and Bala and the factors that
enhance both.
LITERARY REVIEW
The word „vyadhikshamatva’ is made of
two words- „vyadhi + kshamatva‟. Vyadhi is a
condition which comes into existence as
consequence of non-equilibrium of the doshas
(physiological factors i.e vata, pitta & kapha),
dhatus (tissues systems) and malas (excretory
products of body) which in their normal status
maintains the physical and psychological health
(Dass Ranjip Kumar, 2013). The other word
„kshamatva’ is derived from „kshamus sahane’
meaning there by to be patient or composed to
suppress anger, to keep quite or to resist. So,
vyadhikshamatwa is that factor which limits the
pathogenesis and opposes the strength of
disease.
There are nine factors in an individual
which promotes them towards the incapability
to resist the disease manifestation (Trikamji
Yadavaji, 2005).
1) Ati- Sthoola (Excessively obese persons)
2) Ati-Krisha (Excessively emaciated person)
3) Anivista-Mamsa (Individual having
improper musculature)
4) Anivista-Asthi (persons having defective
bone tissues)
5) Anivista-Shonita ( persons with defective
blood)
6) Durbala (Constantly weak person)
7) Asatmya-Aaharopachit (Those nourished
with unwholesome food)
8) Alpa-Aaharopachit (Those taking diet in
small quantity) 9) Alpa-Sattva (Individuals with
feeble mind)
Along with this, there are some parameters
which make the body resistant to the external
environmental changes. Individuals whose
bodies are neither too corpulent nor too
emaciated, who possess healthy and good
musculature, blood and bones are well
nourished with whole some and sufficient food
and who have strong manas are capable of
resisting diseases. Persons having proportionate
musculature, compactness of the body and
healthy sensory & motor organs are not
overruled by the onslaught of diseases. They
can withstand hunger, thirst and the heat of the
sun, cold weather and physical exertion. They
can also digest and assimilate food properly.
The factors which enhance the status of
vyadhikshamatva (Trikamji Yadavaji, 2005).
These are mentioned below-
1) Adequate Ojas
2) Adequate Bala
3) Follow of Seasonal Diet & regimen
4) Proper Anupana (post prandial drinks)
5) Consumption of six Rasa (type of foods)
6) Jatakarma (Birth ceremonial rituals)
7) Drugs (antioxidants)
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When etiological factors come in contact
with a person then they try to produce diseases.
Some persons remain unaffected even after
coming in contact with the relevant pathogenic
factors while some others become victims of
the disease. This capacity or power which
makes the body capable to fight against the
presentation of diseases in the body is said as
vyadhikshamatva. The reasons for the same is
given by Charaka (200 BC) that people have
different ability to resist the disease causing
factors. The reason is that when the resistance
power of the body is sufficiently strong it
destroys the causes. A conference titled
“factors which provoke the diseases” the
conclusion is that “apathya aahara (unhealthy
food) is the cause for diseases and pathya
aahara (healthy food) is cause for health”.
Here, the significance of favourable and non-
favourable diet in the development and
production of a disease is proved. (Trikamji
Yadavaji, 2005)
It is also known that many people with right
daily and seasonal habits also develop disease
and some though indulging in improper habits
hardly get any disease. Therefore a search for
the answer to above conditions lead to some
more factors involved in disease manifestation
and the individual‟s body provides a fertile bed
for the disease causing agents. The disease not
only depends upon the etiological factor but
also depends on the resistance of the individual.
So we can have a greater safety from the
disease if we possess a good health (Deha
bala).
The concept of vyadhikshamatva described
in charaka samhita (1500 BC) gives us a detail
account of immunology and gives us newer
thoughts to present day modern medical
immunology.
Caraka (200 B.C) quoted that all the
individuals have different resistance for
diseases. Vyadhikshamatva (the functional
ability of immune system) varies from
individual to individual in spite of their similar
nutritional, environmental, physical and mental
status.
Chakrapani (Ayurveda-Deepika, 1100
A.D) commented on the term vyadhikshamatva
and gave two opinions:
(I) Vyadhi-balavirodhitvam: It is the capacity
to restrain or withstand the strength
(severity) of the diseases i.e., strength to
arrest the progress of disease.
(II) Vyadhi-utpadapratibandhakatva: The
resisting power of the body competent
enough to arrest the occurrence and re-
occurrence of the disease.
These both sub-types of vyadhikshamatva
commutatively form the resistance which now
a day‟s called Immunity that is specific and non
specific resistance.
DISCUSSION
Excessively obese or emaciated persons are
very weak i.e. they are durbala. Similarly, the
strength of the body depends largely upon the
status of dhatus i.e. mamsa dhatu, asthi dhatu
and sonita. Hence persons in whom these three
dhatus have improper physiological functions
are naturally very weak. Caraka (200 BC) has
especially mentioned mamsa, asthi and sonita
have direct relation with healthy immune
response. This may be due to following
reasons.
A. The healthy condition of these three dhatus
will naturally maintained on equilibrium of
three doshas, which is also required for
resisting disease. This is because there is
relation of interdependence (ashraya-
ashrayee-bhava) between rakta and pitta,
mamsa and kapha and asthi and vata.
B. The cells; WBC of blood (rakta) fight
against infections and prevent the body
from foreign pathogenic substance.
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C. The red bone marrow in the asthi (bone) is
responsible for manufacture of the red and
white cells of the blood.
Persons involved with alpa (inadequate)
and ahita aahara (unfavourable food) are very
weak, as their dhatus (tissue system) are not
properly nourished.
Persons having feeble sattva (mind) are
also weak because sarirabala (Physical
strength) and sattvabala (Psychological
strength) are interdependent so, the weak
manas result in the weak sarira resulting poor
immunity.
Persons having good bala are found with
good defense mechanism. It is the bala which
makes the person capable for performing all the
physical and psychological actions. The proper
psychosomatic status of the body has adequate
agnibala (digestive capacity), dehabala
(physical strength) which manifests itself as
healthy immune system.
Kala (time) also affects the immunity the
visarga kala (southern solstice) increases the
bala of the body results in healthy immunity
Food taken in proper quality and quantity helps
in bringing the strength, complexion, happiness
and longevity. The habit of using all the six
rasa everyday is ideal for maintenance of
health. Intake of madhura (sweet) and amla
(sour) rasa (taste) produces more strength in
the body. To avoid the adverse affects of
dosha’s caya, prakop and prasamana one
should have to follow the seasonal regimens by
consumption of the rasa which are opposite to
kalakrat caya, prakop of doshas (Sharma
Shivprasad, 2008).
Licking of madhu (honey), ghrita (fat) in
unequal quantity with gold at the time of birth
to neonates act as an external nutrition for them
to activate their digestive system. Gold has a
property to enhance the immune system
(Shastri Ambikadutta, 2003).
Jatakarma Karnavedhana sanskara
(ceremonial rituals of ear puncturing) is done at
6th
or 7th
month of age by puncturing the ear
pinna for raksha (protection or resistance
power) and bhusan (cosmetic value). Several
drugs are claimed to enhance immunity i.e.
before gramya dharma (coupulation) tail
pradhana ahara for female and ghrita
pradhana ahara for male is prescribed.
Specific drugs for pregnant women are
described in different months of pregnancy for
better fetal growth. According to the age group
of children different aahara kalpana are
prescribed for enhancing bala (strength) of
children. Rasayana therapies which fortify
dhatus are described in all the texts for
enhancing bala and prevent old age and
disease. (Tripathi Brahmanand, 2010)
VYADHIKSHAMATVA & BALA
Bala is a multidimensional functional
identity of the body, which is the output of
proper equilibrium state of dosha, dhatu &
mala in the body. This comes in the form of
ojas which is considered as the supreme
essence of all the seven rasadi dhatus. It is also
stated as sarvadhatusara. The elementary
constituents of our body i.e. rasa, rakta,
mamsa, meda, asthi, majja & sukra contribute
their extreme purest extract in the formation of
ojas. In this manner ojas is formed in our body
which is the purest factor of all the dhatus
(Flow Chart 1).
The expression of this supreme essence in
the body results in the good musculature, stable
body, ability to perform activities, clarity of
voice, good complexion, healthy karmendriyas
(motor organs and nerves) & gyanendriyas
(sensory organs and nerves) and healthy
vyadhikshamatva (immunity power) of body.
All these features are related to bala. Hence, it
is to be said that bala is the karya (action) and
ojas is the karan (cause) for bala.
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 386–391
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
FLOW CHART 1: Shows the relationship of Bala with other bodily factors
Bala is visualized as an identity which is
observed as the following:
1. Adaptability of the body to the climate with
adequate quantity & quality of rasadi
dhatus. Which are externally observed as
well built body, stable musculature and to
carry out physical activities in a normal
way.
2. Psychological stability in life which is
observed as the person endowed with
memory & devotion, grateful, learned,
pure, courageous, skill-full, resolute,
fighting in battles with prowess, free
from anxiety, intellectual engaged in
virtuous acts.
3. Specific resistance in comparison to
specific diseases which is otherwise called
as pratyanek bala or immunity.
CONCLUSION
A person with proper amount of bala is also
said to be complete with adequate amount of
saptdhatusara (tissue system). This provides
the capacity to resist the external disease
causing agents. It is concluded that
vyadhikshamatva is the external and internal
manifestation of bala (vital strength). The
kapha prakriti body constitution, possess the
best sahaja (natural) bala when compared to
pitta and vata body constitution persons.
REFERENCES
Shastri Ambikadutta (2003) - Sushruta
Samhita, Hindi Commentary
Ayurvedatatva-Sandeepika by
Ambikadutt Shastri 4th
edition,
Chowkhamba Sanskrit Sansthan,
Varanasi.
Trikamji Yadavaji (2005) – Charaka Samhita,
Sanskrit Commentary Ayurveda-Dipika
by Chakrapanidutta Edited by Vaidya
Yadavaji Trikamji Acharya,
Chaukhamba Surabharti Prakshan,
Varanasi.
Sharma Shivprasad (2008) - AstangaSamgraha,
with the Sanskrit Commentaries,
Sashilekha of Indui, Edited by
Dr.Shivprasad Sharma, Chaukhambha
Sanskrit Series Office, Varanasi.
Jangid Chhagan, Vyas H A, Dwivedi R.R.
(2008), “CONCEPTUAL AND
APPLIED STUDY ON EFFECT OF
RITUS ON BALA AS PER THE
SUTRA – Adavante Ca – Nirdiset”
IPGT & RA, Jamnagar.
RASA
RAKTA
MAMSA
MEDA
ASTHI
MAJJA
SUKRA
SUPREME
EXTRACT
OJAS
(karan)
BALA
HEALTHY PHYSIQUE
HEALTHY IMMUNITY HE
AL
TH
Y M
IND
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 386–391
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
Dass Ranjip Kumar (2013), THE ROLE OF
PANCHAKARMA THERAPY IN
MUSCULOSKELETAL DISORDERS
WITH SPECIAL REFERENCE TO
VATAVYADHI, Global J Res. Med.
Plants & Indigen. Med., Volume 2(1):
23–29
Tripathi Brahmanand (2010) – Sharangdhar
Samhita, Amatel with Dipika hindi
commentary by Brahmanand tripathi,
Chaukhamba Surbharti Prakashan,
Varanasi.
Source of Support: Nil Conflict of Interest: None Declared
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 392–400
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
A REVIEW ON VARIETIES OF ARKA - CALOTROPIS PROCERA (AITON)
DRYAND. AND CALOTROPIS GIGANTEA (L.) DRYAND.
Poonam1*, Gaurav Punia
2
1Assistant Professor, Gaur Brahmin Ayurvedic College, Rohtak., India
2HCDS Dental Surgeon, Khanda kheri, Hisar.
*Corresponding Author: E-mail: [email protected], [email protected],
[email protected]; Mob: +918930123462, +9198930123430
Received: 14/03/2013; Revised: 14/04/2013; Accepted: 23/04/2013
ABSTRACT
Herbal medicines have been used from the earliest times to the present day. The ethno-botanical
pharmacology is as old as man himself. Herbal medicines exhibit a remarkable therapeutic diversity.
Arka is a plant which has been used in several traditional medicines to treat a variety of diseases.
Calotropis procera and Calotropis gigantea are two varieties of Arka described in Ayurveda by the
name of Rakta Arka and Shveta Arka. Both have almost similar properties but C. procera is
considered somewhat superior. In Ayurvedic texts also by the name of Arka, Acharyas mentioned
Rakta Arka (Calotropis procera) because of having more medicinal properties than C. gigantea. This
plant has been known to possess analgesic, antitumor, antihelmintic, antioxidant, hepatoprotective,
antidiarrhoeal, anticonvulsant, antimicrobial, oestrogenic, antinociceptive, and antimalarial activity.
KEY WORDS: Rakta Arka, Calotropis gigantia, Arka, antinociceptive.
Review article
Cite this article:
Poonam, Gaurav Punia (2013), A REVIEW ON VARIETIES OF ARKA - CALOTROPIS PROCERA
(AITON) DRYAND. AND CALOTROPIS GIGANTEA (L.) DRYAND., Global J Res. Med. Plants &
Indigen. Med., Volume 2(5): 392–400
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 392–400
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
INTRODUCTION:
Arka (Rakta Arka) an important drug of
Ayurveda is known since Vedic period. In the
vedic literature Arka leaves were used in the
sacrifical rites (Sharma P.V., 2005). There are
two common species of Calotropis, viz
Calotropis gigantea (L.) Dryand nd C. procera
(Aiton) Dryand described in the classical
literature of Ayurveda by the name of Shveta
Arka and Rakta Arka. Calotropis is a genus of
plants that produce milky sap hence also
commonly called milkweed. The latex of
Calotropis procera is said to have a mercury-
like effects on the human body, and is
sometimes referred to as vegetable mercury and
is used in place of mercury in aphrodisiacs. C.
procera is used variously but sometimes leaves
are fried in oil for medicinal purposes. Cattle
often stay away from both the plant C. procera
and C. gigantea because of their unpleasant
taste and due to presence of Cardiac Glycosides
in its sap. Root bark of C. procera has Digitalis
like effect on the heart, but was earlier used as
a substitute of Ipecacuanha. It is a poisonous
plant; calotropin, a compound in the latex, is
more toxic than strychnine.
The Calotropis Shrub that produces white
or sometimes voilet flowers is called as
Swetarka. This is a rare shrub. Flowers of
Swetarka are considered to be favorite of Lord
Shiva.
C. gigantea as indicated by its name it is
much larger and coarse plant than C.procera.
Hindus obtain Ganapati from the root of C.
gigantea that sometimes takes the shape of
Lord Ganesh. The root of C. gigantea shrub is
invited on some auspicious day (in the Ravi-
Pushya Nakshatra ) and carved into the form of
Ganapati or Ganesh in some auspicious
muhurta. Hindus believe that those who
worship this idol of Ganesh enjoy the presence
of Mahadevi Laxmi and Lord Shiva. Most
recently C. gigantea is scientifically reported
for several medicinal properties viz. the flowers
are reported to possess analgesic activity,
antimicrobial and cytotoxic activity. Leaves
and aerial parts of the plant are reported for
anti-diarrhoeal activity, anti-Candida activity
and antibacterial activity, antioxidant activity.
Roots are reported to contain anti-pyretic
activity, cytotoxic activity. Although both
varieties of Arka have almost similar properties
yet C. procera is having comparatively more
medicinal properties.
Synonyms of Arka:
Shveta Arka Ganrupa, Mandar, Vasukh, Shvetapushpa Sadapushpa, Alarka,
Partapsh
Rakta Arka Arkaparna, Vikran, Raktapushpa, Sukhalphal, Ashphot. All
synonyms of Sun belong to Arka, because it has tikshnata like that
of Sun.
English Name Milk weed, Sodom Apple (Deseret), Madar, Giant milkweed
Marathi Rui, Akda
Bangla Akanda
Gujarati Akdo,
Due to its poisonous activities it is called as Darkhatae Jahnak.
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 392–400
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
Sources of Arka
Raktaarka – mainly has two sources
Calotropis procera – smaller red flowers -
(Chief source of Raktaarka in N. India).
Calotropis gigantea (red variety) – larger red
flowers – (Chief source of Raktaarka in S.
India) (Bodhi Nighantu, 2010)
Shwetaarka – mainly has only one source
Calotropis gigantea (White variety) – larger
White flowers – (Chief source of Shwetaarka
all over the country). (Bodhi Nighantu, 2010).
Most of the Nighantus (Ayurvedic Materia
medica) have mentioned same properties of
both Arka (Sharma Ramprasad, 1990). Dalhana
in commentary Of Susruta Samhita mentioned
that by Arka we should take Rakta Arka. Rakta
Arka is having more tikshnata (having more
sharp in properties) compared to Shveta Arka
(Vyas Shiv Kumar, 1986).
Habitat
Calotropis procera is native to northern
Africa (i.e. Algeria, Egypt, Libya, Morocco,
Eritrea, Ethiopia, Somalia, Sudan, Kenya,
Tanzania, Uganda, Cameroon and Equatorial
Guinea)
Calotropis gigantea (Crown flower) is a
species of Calotropis native to Cambodia,
Indonesia, Malaysia, Philippines, Thailand, Sri
Lanka, India and China.
The second variety is of 2 types ie. C.
gigantea (Red) & C. gigantea (White).
C. procera Grown in Punjab, Bihar,
Mumbai. The white variety of Calotropis
gigantea is indigenous to Rajasthan & Gujarat
(which is taken as Shwetarka in Ayurveda)
The red variety of Calotropis gigantea is
common in South India.
So the white variety of C. gigantea has
been introduced into South India especially in
temples for its sanctity. (but when scaned
through the market samples in South India the
root of this variety is hardly available) (Bodhi
Nighantu, 2010)
Botanical Description:
Calotropis procera (Rakta Arka) is a soft-
wooded, evergreen, perennial shrub grows up
to height of 3–6 m. It has one or a few stems,
few branches, and relatively few leaves, mostly
concentrated near the growing tips. The bark is
corky, furrowed, and light grey. A copious
white sap flows whenever stems or leaves are
cut. Giant milkweed has a very deep, stout
taproot with few or no near-surface lateral
roots. Giant milkweed roots reach up to depths
of 1.7–3.0 m in Indian sandy desert soils
(Sharma B.M., 1968). The opposite leaves are
oblong-obovate to nearly orbicular, short-
pointed to blunt at the apex and have very short
petioles below a nearly clasping, heart-shaped
base (Nadkarni A.K., 2000). The leaf blades are
light to dark green with nearly white veins.
(Rastogi RP, 1999). They are 7–18 cm long and
5–13 cm broad, slightly leathery, and have a
fine coat of soft hairs that rub off.
inflorescenence is panicale with purple corolla
and erect lobes. (Anonymous, 2000). The
flowers are umbelliform cymes that grow at or
near the ends of twigs. Flowers are shallowly
campanulate with five sepals that are 4–5 mm
long, fleshy and variable in colour from white
to pink, often spotted or tinged with purple.
The fruits are inflated, obliquely ovoid follicles
that split and invert when mature to release flat,
brown seeds with a tuft of white hairs at one
end Sodom's apple milkweed produces a
simple, fleshy fruit in a grey-green inflated pod,
containing numerous flat, brown seeds with
tufts of long, white silky hair (‘pappus’) at one
end (Howard, R.A., 1989)
C. gigantea is a large shrub or small tree,
about 4–10 m tall. Its stem is erect, up to 20 cm
in diameter. The leaves are broadly elliptical to
oblong-obovate in shape, with the size of 9–20
cm × 6–12.5 cm but subsessile. The cymes are
5–12.5 cm in diameter. The inflorescence stalk
is 5–12 cm long, the stalk of an individual
flower is 2.5–4 cm long. Sepal lobes are
broadly egg-shaped with a size of 4–6 mm × 2–
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 392–400
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
3 mm. Petal is 2.5–4 cm in diameter. It has
clusters of waxy flowers that are either white or
lavender in colour. Each flower consists of five
pointed petals and a small, elegant "crown"
rising from the centre, which holds the stamens.
The plant has oval, light green leaves and milky
stem. The petal lobes are broadly triangular
measuring 10–15 mm × 5–8 mm; they are pale
lilac and cream coloured towards the tips. The
outgrown like structure from the petal (corona)
has 5 narrow fleshy scales, connected to and
shorter than the staminal column, forming an
upturned horn with 2 obtuse auricles on either
side, cream coloured or lilac to purple, with a
dense longitudinal dorsal row of short white
hairs. The egg-shaped or boat-shaped fruits are
mostly in pairs, inflated, 6.5–10 cm × 3–5 cm.
The flowers last long. Taxonomical comparison
between C. procera & C. gigantea is given in
Table. No 1.
Table No 1- Comparison between C. procera and C. gigantea
Plant Origin Height Leaves Flower Petiole Fruits
Calotropis
gigantea/
Gigantic swallow
wort,
Madar/Shveta
Arka
India 8–10
m
Opposite White to purple,
rarely light
green yellow or
white. Flowers
not scented
Sessile Follicles recurved, 2
or 1 follicles,
second more often
suppressed, 3–4"
long
Calotropis
procera/Rakta
Arka
Swallow-
wort/Rakta Arka
India 3–6 m Opposite White to pink,
scented.
Sub-
sessile
Follicles 3–4",
recurved
The photosynthetic capacity of C.
procera is higher than that of C. gigantea
(Wilmer Tezara et al., 2011).
Properties of both arka according to
Bhavprakash (Chunekar K.C., 2007)
Shveta Arka Pushpa-Vrishya (potent), Laghu
(Light), Dipana (Appetizer), Pachna
(Digestive), Aruchi, Prasek (controls excessive
salivation), Svasa Kasa hara (cures asthma and
cough)
Rakta Arka: Madhura Tikta, krimi (removes
worms) kushta (cures all type skin disease)
kapha hara arsha (piles), visha (poison),
raktapitta (haemophillia), gulma, sopha hara
(removes inflammation).
Chemical compostion-
Calotropis procera plant latex contains the
cardenolide, proceraenin, while the root bark
contains benzoylinesolone and
benzoylisolinelone. The leaves and stalk
contain calotropin, and calotropagenin while
the flower contains calotropenyl acetate, and
multiflavenol and the latex contains uzarigenin,
and terpenol ester (Yoganarasimhan SN, 2000).
Chemical investigation of this plant has shown
the presence of triterpenoids, calotropursenyl
acetate and calopfriedelenyl, a norditerpenyl
ester, calotropternyl ester oleanene triterpenes
like calotropoleanyl ester, procerleanol A and B
(Ansari SH et al., 2001) and cardiac glycosides
calotropogenin, calotropin, uscharin, calotoxin
and calactin(Ahmed KKM et al., 2005). The
plant also has been investigated for the
presence of cardenolides (Seiber JN, 1982) and
anthocyanins. Phytochemical investigation of
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 392–400
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the roots of Calotropis procera yields two new
phytoconstituents, procerursenyl acetate and
proceranol, together with the known
compounds N-dotriacont-6-ene, glyceryl mono-
oleolyl-2-phosphate, methyl myrisate, methyl
behenate and glyceryl-1, 2-dicapriate-3-
phosphate. In the leaves, mudarine is the
principal active constituent as well as a bitter
yellow acid, resin and 3 toxic glycosides
calotropin, uscharin and calotoxin. The latex
contains a powerful bacteriolytic enzyme, a
very toxic glycoside calactin (the concentration
of which is increased following insect or
grasshopper attack as a defense mechanism),
calotropin D I, calotropin D II, calotropin F I,
calotropin F II and a non toxic protealytic
enzyme calotropin (2–3%). This calotropin is
more proteolytic than papain, and bromelain
coagulates milk, digests meat, gelatin and
casein. The whole plant of Calotropis gigantea
contains a- and b- amyrin, teraxasterol,
gigantin, giganteol. They are poisonous plants;
calotropin, a compound in the latex, is more
toxic than strychnine. Calotropin is similar in
structure to two cardiac glycosides which are
responsible for the cytotoxicity of Apocynum
cannabinum. Extracts from the flowers
of Rakta Arka (C. procera) have shown strong
cytotoxic activity in the patients of colorectal
cancer. Quercetin-3-rutinoside is identified in
the roots, stem, leaves, flowers and latex.
Voruscharin is isolated from African plant.
Cardenolides contents in leaf (2.04 mg/gm) and
in latex (162.0 mg/g), mostly calotropagenin –
derived cardenolides present from Calotropis
gigantea, two triterpene esters – 3
methylbutanoates of amyrin and taraxasterol
are isolated from latex of C. gigantea.
calotropins D1 and D2 had been isolated
from C. gigantea (Pal and Sinha, 1980). The
new oxiopregnane- oligoglycosides named
Calotropis A and B have been isolated from the
root of C. gigantea and their chemical structure
have been elucidated by chemical and
spectroscopy methods (Kitagawa Isao, 1992).
The cytotoxic principles of 'Akond mul' (Root
of C. gigantea) cardenoloids glycosides,
calotropin frugoside and 4-O-Beta-D-
glucopyranosyl frugoside were obtained as the
cytotoxic principles (Kiuchi, F., 1998).
Gigantin is a chemical extracted only from C.
gigantea.
Rasa Panchaka :
Rasa Katu, Tikta
Guna Laghu, Ruksha, Tikshna
Virya Ushna
Vipaka Katu
Doshakarma Kapha Vata hara
Uses: All the parts, viz, root, stem, leaf and
flowers of Calotropis are in common use in
indigenous system of medicine. Rakta Arka
(Ait.) R. Br., a wild growing plant of family
Apocynaceae is well known for its medicinal
properties. Different parts of this plant have
been reported to exhibit anti-inflammatory,
analgesic, and antioxidant properties. (Ahmed
UAM, 2006). Important factors of the various
parts of this plant have been widely reported.
Latex has been used in leprosy, eczema,
inflammation, cutaneous infections, syphilis,
malarial and low hectic fevers and as
abortifacient (Basu A, 1997). Rakta Arka latex
demonstrated strong inhibitory effect on E.
coli, S. aureus, S. pyogenes, S. pneumonia,
fungus like Aspergillus niger and yeast
Candida albicans. Efficacy of Rakta Arka in
the treatment of S. typhi and S. paratyphi has
been confirmed. C. gigantea's latex which is
highly poisonous is used as purgative. Also
when thorn entered to foot after removing the
thorn its milk applied to reduce the pain.
Ethanol extract of stems of C. gigantea was
reported for hepato-protective activity in male
Wistar rats against carbon tetrachloride induced
liver damage and showed marked hepato-
protective results. Proteins present in the latex
of C. gigantea are strongly proteolytic and
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 392–400
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responsible for procoagulant activity of C.
gigantea (Rajesh R. 2005).
Leaves: In rheumatism, as an anti-
inflammatory and antimicrobial. The leaves
of Rakta Arka are said to be valuable as an
antidote for snake bite, sinus fistula,
rheumatism, mumps, burn injuries, and body
pain. The leaves of Rakta Arka are also used to
treat jaundice. In Egypt, the dried leaves are
smoked in pipes for relieving cough. A
decoction of the leaves is given to relieve
cough. For whooping cough the leaves are
boiled with Momordica charantia and the
decoction is drank a glassful each time. Leaves
of C. gigantea were reported to carry
antioxidant activity. (Singh N., 2010)
Roots: As hepatoprotective agents against
colds, coughs and elephantiasis, as an anti-
inflammatory, analgesic, anti-malarial and
antimicrobial. The root is also used in the
treatment of venereal diseases like gonorrhea
and syphilis. In these cases the root or root bark
is fermented in honey and given to the patient
to drink, reported that Rakta Arka has anti-
HIV-1. It is therefore a plant of interest for the
treatment of HIV-AIDS. (Remya Mohanraj,
2010) For poisonous snake bites, 2 to 4 leaves
of C. gigantea plant is chewed well by the
patient. Also fresh root of this plant are crushed
well and applied well by rubbing firmly over
the bitten area. The anti-pyretic activity of C.
gigantea roots in water : ethanol (50 : 50) has
been reported (Chitema et al., 2005). Anti-
pyretic activity was studied by using yeast and
TAB (Typhoid) vaccine induced pyrexia in
Albino Swiss rats and rabbits.
Flower: As cytostatic, abortifacient,
antimalarial, in asthma and piles and villagers
in Bikaner district ingest almost all plant parts
of Calotropis procera in various dietary
combinations for malarial fevers and pyrexias. In small doses, powdered flowers of Calotropis
gigantea are useful in the treatment of colds,
coughs, asthma, catarrh, indigestion and loss of
appetite (A. K. K. Mueen, 2005) The flowers
are bitter, digestive, astringent, stomachic,
anthelmintic, and tonic (Agharkar 1991;
Warrier et al., 1996). The alcoholic extract of
the flowers of C. gigantea was reported for
analgesic activity in chemical and thermal
models in mice (Pathak AK, 2007).
Latex: Spectrum of ocular toxicity following
accidental inoculation of latex of Calotropis
procera has been reported which leads to
diminish vision (Samar K Basak, 2009). The
latex also used to induce abortion, infanticide.
Latex also has wound healing properties
(Narendranalwaya, 2009). The anti-
inflammatory property of the latex of Rakta
Arka was studied on carrageenin- and formalin-
induced rat paw oedema model. A single dose
of the aqueous suspension of the dried latex
was effective to a significant level against the
acute inflammatory response. Dried latex and
chloroform extract of roots has been reported to
possess anti-inflammatory activity (Raman
Sehgal, 2005), (Kumar VL, 1994).
Calotropis gigantea yields a durable fibre
useful for ropes, carpets, fishing nets and
sewing.
Adverse effects:
The adverse effects Calotropis procera
consumption are reported to cause blisters,
lesions and eruptions when taken by patients
for the treatment of joint pains and
gastrointestinal problems and ocular toxicity.
Besides sometimes cardio-protective steroid
also show toxicity. The preparations of
Calotropis procera need to be used under the
supervision of a trained medical practitioner
(Lewis Nelson, 2007). Latex of C. gigantea
causes irritation to mucosa. An unidentified
allergen is found in the latex of C. gigantea.
CONCLUSION:
The World Health Organization has
estimated more than 80% of the world’s
population in developing countries depends
primarily on herbal medicines for their basic
healthcare needs. In recent years, traditional
uses of natural compounds, especially those of
plant origin, have received much attention of
the world as they are well known for their
efficacy and are generally believed to be safe
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 5 | May 2013 | 392–400
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
for human use. It is best to use the classical
approach in the search for new molecules to
manage a variety of diseases. A thorough
review of the published literature on Calotropis
procera and C. gigantea shows that it is a
popular remedy in a variety of ethnic groups, as
well as Ayurvedic and traditional practitioners
for the treatment of a range of ailments.
Researchers are exploring the therapeutic
potential of Arka as it is likely to have more
therapeutic properties than known. As in
advanced researches on Calotropis procera has
been proven to be a good medicine in case of
HIV, breast cancer, syphilis etc. which are
becoming challenging to our society.
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Source of Support: Nil Conflict of Interest: None Declared
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