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PHARMACOGNOSTICAL AND EXPERIMENTAL EVALUATION OF
MALLIKA (JASMINUM SAMBAC (LINN.) AIT.) WITH SPECIAL
EMPHASIS ON BREAST CANCER CELL LINE
1*Dr. Priya Pramod Wadate
1Assistant Professor, Department of Dravyaguna Vigyana,
Bhartiya Sanskriti Darshan Trusts College of Ayurved, Wagholi, Pune, Maharashtra, India.
ABSTRACT
In the plant kingdom, there are number of plants that yield medicines
useful to mankind. Jasminum sambac (Linn.) Ait. Family – Oleaceae,
called Mallika in Sanskrit is well known glabrous twining shrub widely
grown in gardens throughout the India. Breast cancer is a type of
cancer originating from breast tissue, most commonly from the inner
lining of milk ducts or the lobules that supply the ducts with
milk.According to V.G.Desai and Kirtikar & Basu Mallika acts on the
female reproductive organs (uterus and breast). The flowers of Mallika
act as „Lactifuge‟, at the puerperal state, in the case of breast abscess.
J. sambac flowers and leaves of Mallika are largely used in folk
medicine to prevent and treat breast cancer. The current in vitro
anticancer study was conducted with an aim to check its anticancer
effect on MCF-7, MDA-MB-231 breast cancer cell lines, at concentrations 10-640μg/ml. The
MTT assay protocol was followed to observe the activity of the study drug. The study drug
was tested in 96 well plates and methanolic and aqueous extract of leaf and flower of mallika
were used in different dose levels. As per MTT assay protocol, methanolic extract had shown
significant activity on MCF-7 breast cancer cell line and had shown negligible activity on
MDA-MB231.
KEY WORDS: Jasminum sambac linn. Ait, breast cancer, MCF-7, MDA-MB-231 and MTT
assay, in vitro study.
WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES
SJIF Impact Factor 7.421
Volume 8, Issue 9, 754-769 Research Article ISSN 2278 – 4357
*Corresponding Author
Dr. Priya Pramod Wadate
Assistant Professor,
Department of Dravyaguna
Vigyana, Bhartiya Sanskriti
Darshan Trusts College of
Ayurved , Wagholi, Pune,
Maharashtra, India.
Article Received on
2 July 2019
Revised on 22 July 2019
Accepted on 12 August 2019
DOI: 10.20959/wjpps20199-14612
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1.0 INTRODUCTION
Todays lifestyle are so hectic & living in such a fast growing and newly emerging world of
modern science, people are suffering from many lifestyle diseases like Alzheimer‟s disease,
Arterisclerosis, Cancer. Cancer refers to a large number of conditions that is characterized by
abnormal cell division.The cells of cancer infiltrate and destroys surrounding healthy tissue.
They have the ability to metastasize and spread throughout the body.
Cancer occurs in all ages of human being. But certain cancers are life threatening for men and
women and for so many women; there is no more deadly disease than breast cancer. Breast
cancer is a type of cancer originating from breast tissue, most commonly from the inner
lining of milk ducts or the lobules that supply the ducts with milk.
Jasminum sambac (Linn). Ait Family – Oleaceae, called Mallika in Sanskrit. It is well known
glabrous twining shrub widely grown in gardens throughout the India. As per the paper
published on antitumor activity of J. sambac flowers and leaves of Mallika are largely used in
folk medicine to prevent and treat breast cancer. Flowers are useful to women when brewed
as a tonic as it aids in preventing breast cancer and stopping uterine bleeding.
1.1 SELECTION OF DRUG
According to V.G.Desai and Kirtikar & Basu Mallika acts on the female reproductive organs
(uterus and breast). The flowers of Mallika act as „Lactifuge‟, at the puerperal state, in the
case of breast abscess.
More than 30 ayurvedic medicinal plants like Amruta, Haridra, Yashtimadhu, Kanchnar,
Guggulu etc. have proved the anticancer activity on different cancers. The species of
jasminum i.e. J. grandiflorum i.e. Jati had already studied for anticancer activity by cell line
study on MCF 7 and MDA MB231.
Sushrutacharya stated that “Rakta” is one of the “Dushya” for Vidradhi (Abscess) and
Granthi (Cyst) formation. In the Susrut samhita Mallika is described as , ”Pittanashini” and in
Raj Nighantu it described as “Vranaropani” , ”Pittanashini” , “Pittaprakophrut”. Due to
“Ashraya-ashrayi Bhav” of Pitta and Rakta Mallika also acts as “Raktdushtihar”. So it can be
useful in the Arbuda and Granthi chikitsa.
Jasminum sambac is studied for antitumor activity by animal study and cervical cancer cell
line. Other species of Jasminum like jasminum grandiflorum have proved their anticancer
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activity by experimental study but still the work on anticancer activity of Mallika on Breast
cancer cell line is not found. So the further attempt has been made to scientifically evaluate
the anticancer activity on breast cancer cell line.
1.2 NEED OF STUDY
Modern science is yet to discover effective remedies for deadly diseases. These diseases are
very complicated in nature with complexity at every level–Anatomy, Physiology,
Biochemistry, Molecular biology, and Gene expression. So treating such a disease is a big
challenge, Worldwide, breast cancer comprises 22.9% of all cancers (excluding non-
melanoma skin cancers) in women. In 2008, breast cancer caused 4,58,503 deaths worldwide
(13.7% of cancer deaths in women).
Additionally toxicity of the drug, major surgeries , radiation damage , chemotherapy hazards
make the situation worse. Survival rates in the Western world are high; for example, more
than 8 out of 10 women (84%) in England diagnosed with breast cancer survive for at least 5
years. In developing countries, however, survival rates are much poorer.
The excruciating experience of dying cancer petient can be ameliorated by making use of
Ayurveda. Ayurveda can be helpful in the management of cancer in many ways such as
prophylactic, palliative, curative, supportive.
1.3 AIM & OBJECTIVES
AIM - To assess the anticancer activity of mallika with special emphasis on breast cancer cell
line by using MTT assay.
OBJECTIVES
To asses the identity of Mallika through pharmacognostical studies.
To extract the phyto-constituents of flowers and leaves of Mallika.
To evaluate anti cancerous activity by Cell based study by performing various assay.
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1.4 DRUG REVIEW
1.4.1 References of Mallika in Bruhtrayee
Table 1: References of Mallika from Bruhatrayee.
Names of sthan Charak samhita Sushrut samhita Ashtang hrudaya
Sutra - 1 2
Nidan - - -
Viman - - -
Sharir - - -
Indrya - - -
Chikitsa - - 1
Kalp - - -
Sidhi - - -
Uttartantra - - -
Utarsthana - - -
Total ref - 1 3
1.4.2 SYNONYMS
Synonyms are the word with same or similar meanings. In Ayurvedic text it holds its own
importance as it describes the morphological and pharmacological aspect of the drug.The
synonym of Mallika explained in ancient text are given below.
Table 2: Synonyms of Mallika in Nighantu.
Synonyms DN RN MN KN SHN AN BPN NA API
Asphota + +
Ashtapadi + +
Gandhanamka +
Gouri +
Girija +
Gavakshi + +
Chandrika +
Jati +
Damayanti +
Narishtha +
Pushpati +
Pramodani + + +
Priya + +
Bhadravalli + +
Bhoopadi + + + +
Madaniya + +
Madayanti + + + + +
Mallika + + + + + + + +
Malati +
Muktabandhana + +
Modini + +
Trunashunya + +
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Yuthika +
Varshika + +
Vanchandrika +
Vichakila +
Sheetbhiru + + + + + +
Soumya + +
1.4.3 GUNA – KARMA
Rasa, guna, veerya, vipak, and prabhava which are simplest parameters to ascertain the
actions of the drugs, are considered as the base of the pharmacology in Ayurveda. The
pharmacological properties of Mallika as mentioned by different acharyas, are presented in
following.
Table 3: Guna Karma of Mallika.
Guna SS DN RN MN KN BPN SHN SHAN NA API
RAS Katu + + + + + + +
Tikat + + + + + + +
Swadu + +
GUNA Laghu + + + +
VEERYA Ushna + + + +
Sheet +
VIPAKA Madhur +
KARMA
VATAHAR + + + +
PITTAHAR + + + + +
KAPHAHAR +
RAKTAJEETA +
VAJIKARAN + +
VRANAHAR + + +
CHAKSHUSHYA + + +
HRUDYA +
TWAKDOSHAHAR + + + +
URDHVAJATRUGAT
VYADHI + +
VISHAHAR + + + + +
KRIMIGHNA +
SHUKRAL +
ARUCHI + + +
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2.0 MATERIAL AND METHODS
2.1 MATERIAL
Drug:-
Drug name – Mallika.
Botanical name - Jasminum sambac Linn. (Ait.)
Family – Oleaceae.
Useful part - flower& leaves.
Source - Anna Hajare Garden,Wadgaon Sheri pune.
Collection - Leaves collected in Varsha rutu flowers collected from plant in Grishmarutu.
Authentication - BOTANICAL SURVEY OF INDIA, PUNE.
2.2 METHODS (STEPS IN EXPERIMENTAL STUDY)
1. Collection & Authentication.
2. Drying.
3. Preparation of Herbal extract.
4. Extraction.
5. Centrifugation.
6. Filtration.
7. Preparation of concentration of extract.
8. Preparation of cell & MTT assay.
2.2.1 DRUG COLLECTION
Flower - May 2015 (Grishma rutu).
Leaf - September 2015 (Varsha rutu).
2.2.2 DRYING
Weight of wet Mallika Leaf - 3kg.
Weight of dry Mallika Leaf - 350gms.
Weight of wet Mallika Flower - 2.5kg.
Weight of dry Mallika Flower- 230gms.
STORAGE - are done as per GMP.
1. Dried samples (leaves and flowers) are stored and handled in a manner which prevents
their mixing up, contamination.
2. Containers of materials are closed, and bagged or boxed.
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3. Containers of materials are labelled with respect to identity.
4. Materials are sampled and tested or examined in conformance.
2.2.1 AUTHENTICATION
Drug has been identified and authentified at BOTANICAL SURVEY OF INDIA, PUNE.
No.BSI/WRC/Tech./2014/.
2.2.3 PREPARATION OF HERBAL EXTRACT
A) Methanolic extract
1. The plant material was ground into coarse powder and 10gm of powder was placed inside
a thimble made from thick filter paper.
2. The thimble was placed into the main chamber of the Soxhlet, extractor.The Soxhlet
extractor was placed into a round bottom flask containing the extraction solvent.
3. The solvent was heated so that the vapours travel up a distillation arm, and flood into the
chamber where the thimble was placed.
4. The chamber containing the thimble fills with warm solvent and depending upon the
solubility different bioactive compounds dissolve in the solvent.
5. When the Soxhlet chamber was almost full, the chamber was automatically emptied by a
siphon side arm, with the solvent running back down to the distillation flask.
6. This cycle was repeated 10-15 times.
7. In this manner extract of leaf and flower were prepared.
B) Aqueous extract
1. The plant material was ground into coarse powder.
2. 30gm powder was placed in the glass beaker.
3. 480 ml water was added in the beaker which was sixteen times than the powder.
4. The solution was heated on the low flame up to the one eighth solution was remaining.
5. The decoction was filtered by muslin cloth.
6. Decoction was centrifuged at 12000 rpm for 15 min.
7. Supernatant solution was kept for overnight drying at 60°C.
8. In this manner aqueous extract of leaf and flower were prepared.
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2.2.4 PREPARATION OF CONCENTRATIONS OF THE EXTRACTS
1. Weighing of herbal extract.
2. Drying it in drier for about 30 to 40 min at 60 °C.
3. Concentration=Final wt-Initial wt.
4. The concentration of methnolic exract of leaf is 4.13gm, flower is 3.2694gm.
5. The concentration of aqueous exract leaf is 3.33 gm, flower is 2.3640gm.
6. Using above value various concentrations of the extracts were prepared (10µg/ml,
20µg/ml up to 640µg/ml) in 10% DMEM.
2.2.5 PREPARATION OF CELL
Maintenance of cell lines: The obtained cell lines were maintained using sterile DMEM
(Gibco) supplemented with 10% FBS (Sigma) at 5% CO2 and 37°C in an incubator
(Thermoforma) in a T-50 flask (Falcon).
Passaging of cells: Cultured cells with adherent property form a monolayer during their
growth. These cells grow laterally to occupy the entire matrix of the culture flask, at this
stage the cells are said to be confluent and for further maintenance passaging was done.
Freezing of cells: Cells which have attained confluency are freezed to preserve them for
further use.
Thawing of cells: Thawing was performed to revive the frozen cell lines. The cryovial
containing cells was retained in warm water prior to transfer.
2.2.6 CYTOTOXICITY DETERMINATION USING MTT ASSAY
Principle
MTT (3- (4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazoliumbromide) assay is performed to
determine the cytotoxic effect of the extract on cell lines. This assay can be used to quantify
the living cells based on the presence of active enzyme systems which is a prerequisite for
live cells.
The MTT dye is reduced to form purple formazan crystals in living cells by various enzymes
of the class oxidoreductase. The formazan crystals are dissolved in solvents like SDS, DMF
etc.
In this experiment the presence of a mitochondrial inner membrane bound enzyme, succinate
dehydrogenase was detected using MTT dye. The presence of this enzyme ensured the
viability of the cells.
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Procedure
Day 1: Seeding of cells in 96 well plate.
Day 2: Dosing of cells in 96 well plate.
Day 3: The media containing the dose was discarded and 100 µl of the MTT solution was
added to every well.
2.3 OBSERVATIONS
2.3.1 Pharmacognostical Study
Standardisation and phyto-chemical analysis of leaf and flower powder is done at IDRAL,
Pune. Report No. 272,273.
Table 4: Organoleptic study.
Sr. No. Characters Leaves Stem Root
1 Sound No significant No significant No significant
2 Touch Smooth Rough Hairy
3 Color Green Greenish brown Light brown
4 Taste Bitter Bitter Bitter
5 Odour Characteristic Characteristic Characteristic
2.3.2 Microscopical study
Leaf- petiole - shows single layered upper and lower epidermis consisting of tubular cells,
covered with striated cuticle; trichomes of two types, non-glandular, uniseriate, 1-5
celled, warty, and with pointed apical cell.
Midrib - cut at basal region shows both upper and lower single layered epidermis,
externally covered with cuticle.
Lamina- shows a dorsi ventral structure, epidermis single layered, externally covered with
cuticle.
Microscpical study of powder
Leaf Powder - Dark green; Palsied tissue in surface view, Stomata with two guard cells,
parenchymatous epidermal hairs multi cellular, spongy cells with chlorophyll, conducting
strands with spiral thickening, cells with brown coloured substance, crystals of calcium.
Flower Powder - Brown coloured; polygonal parenchymatous cells in surface view, few
cells with brown coloured pigment, small pollen grains are seen, no vascular tissue.
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Table 5: Phyto-Chemical Analysis.
Test leaf Flower
pH 4.61% 4.84%
Moisture 7.88% 8.73%
Total Ash 14.12% 10.29%
Acid soluble Ash 13.80% 10.29%
Water soluble Ash 2.34% 6.64%
2.3.3 CALCUTATIONS
(Absorbance of test sample/ Absorbance of control) × 100=% Viability
Table No. 6: Methanolic leaf extract- MDA MB231 (Breast Cancer Cell).
Concentrations 1 2 3 Average %Viability
Control 0.163 0.165 0.159 0.162333 100
10 ug 0.149 0.123 0.139 0.137 84.45929
20 ug 0.159 0.167 0.15 0.158667 97.69925
40 ug 0.239 0.236 0.251 0.242 149.1726
80 ug 0.279 0.291 0.312 0.294 181.2519
160 ug 0.312 0.315 0.326 0.317667 195.7839
320 ug 0.42 0.423 0.427 0.423333 260.8619
640 ug 0.353 0.385 0.361 0.366333 225.6473
Table No. 7: Aqueous leaf extract- MDA MB231 (Breast Cancer Cell).
Concentrations 1 2 3 Average %Viability
Control 0.128 0.14 0.137 0.135 100
10 ug 0.118 0.1 0.124 0.114 84.70901
20 ug 0.126 0.123 0.137 0.128667 95.43155
40 ug 0.162 0.175 0.175 0.170667 126.4332
80 ug 0.21 0.225 0.245 0.226667 167.8696
160 ug 0.252 0.278 0.289 0.273 202.1318
320 ug 0.292 0.318 0.371 0.327 242.0236
640 ug 0.272 0.279 0.334 0.295 218.5271
Table No. 8: Methanolic flower extract- MDA MB231 (Breast Cancer Cell).
Concentrations 1 2 3 Average %Viability
Control 0.168 0.173 0.154 0.165 100
10 ug 0.149 0.157 0.17 0.158667 96.61051
20 ug 0.165 0.185 0.193 0.181 110.1585
40 ug 0.231 0.26 0.258 0.249667 151.7738
80 ug 0.251 0.288 0.254 0.264333 160.2713
160 ug 0.314 0.323 0.321 0.319333 194.0172
320 ug 0.39 0.391 0.387 0.389333 236.4844
640 ug 0.326 0.314 0.307 0.315667 191.6337
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Table No. 9: Aqueous flower extract- MDA MB231 (Breast Cancer Cell).
Concentrations 1 2 3 Average %Viability
Control 0.131 0.152 0.139 0.140667 100
10 ug 0.119 0.111 0.117 0.115667 82.67956
20 ug 0.139 0.131 0.151 0.140333 100.3081
40 ug 0.189 0.216 0.222 0.209 148.6974
80 ug 0.263 0.259 0.29 0.270667 193.2637
160 ug 0.312 0.304 0.304 0.306667 218.9577
320 ug 0.372 0.386 0.422 0.393333 280.5047
640 ug 0.313 0.349 0.432 0.364667 259.776
Table No. 10: Methanolic leaf extract- MCF7 (Breast Cancer Cell).
Concentrations 1 2 3 Average %Viability
Control 0.246 0.228 0.226 0.233333 100
10 ug 0.265 0.267 0.278 0.27 115.9459
20 ug 0.31 0.293 0.309 0.304 130.4169
40 ug 0.259 0.241 0.243 0.247667 106.1695
80 ug 0.246 0.273 0.234 0.251 107.7589
160 ug 0.23 0.197 0.217 0.214667 91.97238
320 ug 0.236 0.211 0.189 0.212 90.70238
640 ug 0.158 0.161 0.12 0.146333 62.64634
Table No. 11: Aqueous leaf extract- MCF7 (Breast Cancer Cell).
Concentrations 1 2 3 Average %Viability
Control 0.273 0.242 0.274 0.263 100
10 ug 0.27 0.27 0.26 0.266667 101.7873
20 ug 0.373 0.361 0.322 0.352 134.4406
40 ug 0.32 0.264 0.276 0.286667 109.0123
80 ug 0.36 0.388 0.385 0.377667 144.2366
160 ug 0.36 0.345 0.32 0.341667 130.4061
320 ug 0.444 0.487 0.402 0.444333 170.1975
640 ug 0.342 0.345 0.375 0.354 134.8993
Table No. 12: Methanolic flower extract- MCF7 (Breast Cancer Cell).
Concentrations 1 2 3 Average %Viability
Control 0.268 0.263 0.275 0.268667 100
10 ug 0.317 0.332 0.334 0.327667 121.9913
20 ug 0.269 0.354 0.313 0.312 116.264
40 ug 0.284 0.303 0.292 0.293 109.1204
80 ug 0.315 0.24 0.245 0.266667 99.29433
160 ug 0.262 0.322 0.303 0.295667 110.1255
320 ug 0.274 0.267 0.208 0.249667 93.13203
640 ug 0.177 0.184 0.163 0.174667 65.09316
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Table No. 13: Aqueous flower extract- MCF7 (Breast Cancer Cell).
Concentrations 1 2 3 Average %Viability
Control 0.268 0.267 0.21 0.248333 100
10 ug 0.255 0.257 0.236 0.249333 101.2616
20 ug 0.328 0.356 0.329 0.337667 137.4627
40 ug 0.326 0.28 0.271 0.292333 118.5194
80 ug 0.351 0.307 0.304 0.320667 130.2378
160 ug 0.37 0.397 0.349 0.372 150.9798
320 ug 0.487 0.482 0.385 0.451333 181.858
640 ug 0.383 0.377 0.312 0.357333 144.2268
Figure No. 1: MTT on MDA MB231 (Breast cancer cell).
Figure No. 2: MTT on MCF7 (Breast cancer cell).
3.0 RESULT AND DISCUSSION
3.1 RESULT
MTT assay was performed to define the optimal concentration at which extract was toxic
to cells.
Aqueous extracts of both leaf and flower were found to be non-toxic upto 640µg/ml in
both breast cancer cell lines.
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Methanolic extract of leaf and flower were found to be toxic on MCF-7 at the higher two
concentrations (320, 640µg/ml) and non-toxic to MDA MB231 cell line as evident from
graph.
3.2 DISCUSSION
As per the references collected from ayurvedic literature, it is observed that sushrutacharya
and Ashtang Hrudaya karas mentioned mallika as name Mallika itself .All the nighantukaras
gave the synonyms of mallika and they included it in differant vargas.
In pharmacognostical and microscopical analysis there is no significant result found. In
phytochemical analysis pH of leaf and flower is nearby same but other values like moisture
total ash values are varies.
From experimental study it is observed that methanolic extracts of mallika have significant
cytotoxicity on MCF7 breast cancer cells and other extracts are not showing such a type of
activity on both MCF7& MDAMB 231 cell lines.
4.0 CONCLUSION
Mallika has the anticancer activity on MCF7 cell line. There is cytotoxic effect found of
methanolic extracts of leaf and flower of mallika in MCF-7 breast cancer cell line at 320 and
640µg/ml.
Further Scope of Study
Further studies need to be conducted to investigate the cytotoxic activity of the extract in
higher concentrations and also in various assays and in other cancer cell lines.
5.0 REFERANCES
1. Acharya Shri RadhaKrishna Prashar, Ayurvedacharya; Sharangdhara samhita, Baidyanath
Ayurved Bhavan Ltd. Nagpur - 9, 4th Edition, 1994.
2. Ambika Dutt Shastri, Bhaishajya Ratnavali (Vidyotini Hindi commentary), Chaukhamba
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3. Bapalal G. Vaidya, Dravyaguna Shastra, University Granth Nirman board, Gujarat rajya
Ahmedabad. 1st Edition, 1972.
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Edition, 1986.
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5. Bapalal Vaidya Nighantu Adarsh Vol I, II; Chaukhambha Bharati Academy, Varanasi,
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compounds Formulations used in Ayurveda and Siddha; 1996.
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13. Dr. A. P. Deshpande, Dravyaguna Vigyan Vol II, Anmol Prakashan, Pune 1st Edition
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Prakashan, Varanasi, 1st Edition, 1998.
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23. Indra Dev Tripathi, Gadanigraha: Vaidya Shodhal, Vidyotini Hindi Commentary,
chaukhamba Sanskrit Series, Varanasi. 1st Edition, 1969.
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