• Hemant Sood Jaypee University of Information Technology
  • Shivam Sharma
  • Arun Sharma
  • Vineet Mehta
  • Rajinder Singh Chauhan
  • Udayabanu Malairaman


Objective: The present study involves hydroalcoholic extraction of Trillium govanianum (Hindi name: Nag chhatri), which is a high-value medicinal
plant found at the altitude of 2500-4000 m. Aiming in this direction, we performed hydrolysis of extract using response surface methodology (RSM)
for optimizing diosgenin content. The extracts were evaluated for cytotoxicity.
Methods: In RSM, the cumulative effect of independent variables including time (minutes), temperature (°C), and solid-liquid ratio (g/ml)
were investigated through central composite design. Cytotoxicity studies of crude rhizome and hydroalcoholic extract were carried out by
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay on three cell lines, viz., MDCK, MCF-7, and MDA-MB-23.
Results: The diosgenin content obtained was 5.9%, which is reported for the first time in T. govanianum. Hydrolyzed extract showed less toxicity in
MDCK (normal cell line) cells but significantly reduced the proliferation of MCF-7 and MDA-MB-231 cancer cells.
Conclusion: Hydrolysis method was optimized by RSM, which proved to be an efficient method for extraction of diosgenin from T. govanianum.
Hydrolyzed extract showed antiproliferative activity on cancer cell lines with minimal effect on normal cells.
Keywords: Trillium govanianum, Diosgenin, Nag chhatri, Hydrolysis, MDCK, MCF-7, MDA-MB-231.


Download data is not yet available.

Author Biography

Hemant Sood, Jaypee University of Information Technology

Dr. Hemant Sood
Assistant Professor(Sr. Grade)
Department of BT and BI
Jaypee University of Information Technology
Distt: Solan
Himachal Pradesh



Dhar U, Manjkhola S, Joshi M, Bhatt A, Bisht A, Joshi M. Current

status and future strategy for development of medicinal plants sector in

Uttaranchal, India. Curr Sci 2002;83(8):956-64.

Uniyal SK, Singh KN, Jamwal P, Lal B. Traditional use of medicinal

plants among the tribal communities of Chhota Bhangal, Western

Himalaya. J Ethnobiol Ethnomed 2006;2:14.

Aswal B, Mehrotra BN. Flora of Lahaul-Spiti. (A Cold Desert in North

West Himalaya). Dehradun: Bishen Singh Mahendra Pal Singh; 1994.

p. 6.

Chauhan NS. Medicinal and Aromatic Plants of Himachal Pradesh.

New Delhi, India: Indus Publishing; 1999.

Fazli FR, Hardman R. The spice, fenugreek (Trigonellla foenumgraecum



commercial varieties









Sci 1968;10(2):66-77.

Raghuram TC, Sharma RD, Sivakumar B, Sahay BK. Effect of

fenugreek seeds on intravenous glucose disposition in non-insulin

dependent diabetic patients. Phytother Res 1994;8(2):83-6.

Rahman S Ur, Ismail M, Shah MR, Iriti M, Shahid M. GC/MS analysis,

free radical scavenging, anticancer and β-glucuronidase inhibitory

activities of Trillium govanianum rhizome. Bangladesh J Pharmacol


Rahman S Ur, Ismail M, Shah MR, Adhikari A, Anis I, Ahmad MS, et

al. Govanoside A, a new steroidal saponin from rhizomes of Trillium

govanianum. Steroids 2015;104:270-5.

Dangi R, Misar A, Tamhankar S, Rao S. Diosgenin content in some

Trigonella species. Indian J Adv Plant Res 2014;1:47-51.

Ireland CR, Schwabe WW, Coursey DG. The occurrence of batatasins

in the Dioscoreaceae. Phytochemistry 1981;20(7):1569-71.

Niño J, Jiménez DA, Mosquera OM, Correa YM. Diosgenin

quantification by HPLC in a Dioscorea polygonoides tuber collection

from Colombian flora. J Braz Chem Soc 2007;18(5):1073-6.

Acharya SN, Thomas JE, Basu SK. Fenugreek, an alternative crop for

semiarid regions of North America. Crop Sci 2008;48(3):841-53.

Zerbino DR, Birney E. Velvet: Algorithms for de novo short read

assembly using de Bruijn graphs. Genome Res 2008;18:821-9.

Šavikin-Fodulović K, Grubišić D, Ćulafić L, Menković N, Ristić M.

Diosgenin and phytosterols content in five callus lines of Dioscorea

balcanica. Plant Sci 1998;135(1):63-7.

Vidyarthi S, Samant S, Sharma P. Dwindling status of Trillium

govanianum Wall. ex D. Don-A case study from Kullu district of

Himachal Pradesh, India. J Med Plants Res 2013;7(8):392-7.

Liu B-L, Tzeng Y-M. Optimization of growth medium for the

production of spores from Bacillus thuringiensis using response surface

methodology. Bioproc Eng 1998;18(6):413-8.

Kumar V, Chauhan RS, Sood H, Tandon C. Cost effective quantification

of picrosides in Picrorhiza kurroa by employing response surface

methodology using HPLC-UV. J Plant Biochem Biotechnol


Le Man H, Behera S, Park H. Optimization of operational parameters

for ethanol production from Korean food waste leachate. Int J Environ

Sci Technol 2010;7(1):157-64.

Xu Q, Shen Y, Wang H, Zhang N, Xu S, Zhang L. Application of

response surface methodology to optimise extraction of flavonoids

from fructus sophorae. Food Chem 2013;138(4):2122-9.

Cacace JE, Mazza G. Optimization of extraction of anthocyanins from

black currants with aqueous ethanol. J Food Sci 2003;68(1):240-8.

Box GE, Wilson KB. On the experimental attainment of optimum

conditions. J R Stat Soc Ser B Methodol 1951;13(1):1-45.

Drapeau D, Sauvaire Y, Blanch HW, Wilke CR. Improvement of

diosgenin yield from Dioscorea deltoidea plant cell cultures by use of a

non-traditional hydrolysis method. Planta Med 1986;(6):474-8.

Baydoun E, Bibi M, Iqbal MA, Wahab AT, Farran D, Smith C, et al.

Microbial transformation of anti-cancer steroid exemestane and

cytotoxicity of its metabolites against cancer cell lines. Chem Cent J


Mosmann T. Rapid colorimetric assay for cellular growth and survival:

Application to proliferation and cytotoxicity assays. J Immunol

Methods 1983;65(1-2):55-63.

Kokate CK, Purohit AP, Gokhale SB. Pharmacognosy. Pune: Nirali

Prakashan; 2005.

Shah HJ, Lele S. Extraction of diosgenin, a bioactive compound from

natural source Dioscorea alata Var purpurae. J Anal Bioanal Tech


Yang H, Yin HW, Wang XW, Li ZH, Shen YP, Jia XB. In situ pressurized

biphase acid hydrolysis, a promising approach to produce bioactive

diosgenin from the tubers of Dioscorea zingiberensis. Pharmacogn Mag


Coursey D, Martin F, Plucknett D, editors. The Past and Future of

The Yarns as Crop Plants. Tropical Root and Tuber Crops Tomorrow

Proceedings of the Second International Symposium on Tropical Root

and Tuber Crops; 1970.

Desai K, Desmukh V, Saoji A. Studies on Diosgenin content of

Balanites aegyptiaca. East Pharm 1978;21:191-3.

Behera KK, Sahoo S, Prusti A. Biochemical quantification of diosgenin

and ascorbic acid from the tubers of different Dioscorea species found

in Orissa. Libyan Agric Res Cent J Int 2010;1(2):123-7.

Behera KK, Sahoo S, Prusti A. Relative agronomic performance of

different Dioscorea species found in different parts of Orissa. Nat Sci


Fazli F, Hardman R. Isolation and characterization of steroids and

other constituents from Trigonella foenum-graecum. Phytochemistry


Melo PS, De Azevedo MB, Zullo MA, Fabrin-Neto JB, Haun M.

Cytotoxicity of the phytosterol diosgenin and its derivatives in

rat cultured hepatocytes and V79 fibroblasts. Hum Exp Toxicol


Liu MJ, Wang Z, Ju Y, Wong RN, Wu QY. Diosgenin induces cell cycle

arrest and apoptosis in human leukemia K562 cells with the disruption

of Ca2+ homeostasis. Cancer Chemother Pharmacol 2005;55(1):79-90.

Raju J, Patlolla JM, Swamy MV, Rao CV. Diosgenin, a steroid saponin

of Trigonella foenum graecum (Fenugreek), inhibits azoxymethaneinduced







rats and





human colon cancer cells. Cancer Epidemiol Biomarkers



Ono M, Sugita F, Shigematsu S, Takamura C, Yoshimitsu H,

Miyashita H, et al. Three new steroid glycosides from the underground

parts of Trillium kamtschaticum. Chem Pharm Bull (Tokyo)


Yokosuka A, Mimaki Y. Steroidal glycosides from the underground

parts of Trillium erectum and their cytotoxic activity. Phytochemistry


Wang J, Zou K, Zhang Y, Liu C, Wu J, Zhou Y, et al. An 18-norspirostanol

saponin with inhibitory action against COX-2 production from the

Asian J Pharm Clin Res, Vol 9, Issue 4, 2016, 386-392

Sharma et al.

underground part of Trillium tschonoskii. Chem Pharm Bull (Tokyo)


Huang W, Zou K, Xiong B. The rhizome of Trillium tschonoskii Maxim.

extract induces apoptosis in human lung cancer cells. Z Naturforschung

C J Biosci 2011;66(9-10):477-84.

Liu K, Liu PC, Liu R, Wu X. Dual AO/EB staining to detect apoptosis

in osteosarcoma cells compared with flow cytometry. Med Sci Monit

Basic Res 2015;21:15-20.



How to Cite

Sood, H., S. Sharma, A. Sharma, V. Mehta, R. S. Chauhan, and U. Malairaman. “EFFICIENT HYDROALCOHOLIC EXTRACTION FOR HIGHEST DIOSGENIN CONTENT FROM TRILLIUM GOVANIANUM (NAG CHHATRI) AND IT’S IN VITRO ANTICANCEROUS ACTIVITY”. Asian Journal of Pharmaceutical and Clinical Research, vol. 9, no. 4, July 2016, pp. 386-92, https://www.innovareacademics.in/journals/index.php/ajpcr/article/view/12588.



Original Article(s)