EFFICIENT HYDROALCOHOLIC EXTRACTION FOR HIGHEST DIOSGENIN CONTENT FROM TRILLIUM GOVANIANUM (NAG CHHATRI) AND IT’S IN VITRO ANTICANCEROUS ACTIVITY

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

Abstract

ABSTRACT
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.

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
Waknaghat
Distt: Solan
Himachal Pradesh
India

References

REFERENCES
1. 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.
2. 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.
3. Aswal B, Mehrotra BN. Flora of Lahaul-Spiti. (A Cold Desert in North
West Himalaya). Dehradun: Bishen Singh Mahendra Pal Singh; 1994.
p. 6.
4. Chauhan NS. Medicinal and Aromatic Plants of Himachal Pradesh.
New Delhi, India: Indus Publishing; 1999.
5. Fazli FR, Hardman R. The spice, fenugreek (Trigonellla foenumgraecum
L):
It’s
commercial varieties
of
seed
as
a
source
of
diosgenin.

Trop
Sci 1968;10(2):66-77.
6. 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.
7. 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
2015;10(3):577-83.
8. 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.
9. Dangi R, Misar A, Tamhankar S, Rao S. Diosgenin content in some
Trigonella species. Indian J Adv Plant Res 2014;1:47-51.
10. Ireland CR, Schwabe WW, Coursey DG. The occurrence of batatasins
in the Dioscoreaceae. Phytochemistry 1981;20(7):1569-71.
11. 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.
12. Acharya SN, Thomas JE, Basu SK. Fenugreek, an alternative crop for
semiarid regions of North America. Crop Sci 2008;48(3):841-53.
13. Zerbino DR, Birney E. Velvet: Algorithms for de novo short read
assembly using de Bruijn graphs. Genome Res 2008;18:821-9.
14. 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.
15. 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.
16. 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.
17. 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
2015;24(4):376-84.
18. 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.
19. 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.
20. Cacace JE, Mazza G. Optimization of extraction of anthocyanins from
black currants with aqueous ethanol. J Food Sci 2003;68(1):240-8.
21. Box GE, Wilson KB. On the experimental attainment of optimum
conditions. J R Stat Soc Ser B Methodol 1951;13(1):1-45.
22. 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.
23. 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
2013;7(1):57.
24. 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.
25. Kokate CK, Purohit AP, Gokhale SB. Pharmacognosy. Pune: Nirali
Prakashan; 2005.
26. Shah HJ, Lele S. Extraction of diosgenin, a bioactive compound from
natural source Dioscorea alata Var purpurae. J Anal Bioanal Tech
2012;3(4):141-4.
27. 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
2015;11(43):636-42.
28. 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.
29. Desai K, Desmukh V, Saoji A. Studies on Diosgenin content of
Balanites aegyptiaca. East Pharm 1978;21:191-3.
30. 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.
31. Behera KK, Sahoo S, Prusti A. Relative agronomic performance of
different Dioscorea species found in different parts of Orissa. Nat Sci
2009;7(3):25-35.
32. Fazli F, Hardman R. Isolation and characterization of steroids and
other constituents from Trigonella foenum-graecum. Phytochemistry
1971;10(10):2497-503.
33. 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
2004;23(10):487-93.
34. 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.
35. Raju J, Patlolla JM, Swamy MV, Rao CV. Diosgenin, a steroid saponin
of Trigonella foenum graecum (Fenugreek), inhibits azoxymethaneinduced
aberrant
crypt
foci
formation
in
F344
rats and
induces
apoptosis

in
HT-29
human colon cancer cells. Cancer Epidemiol Biomarkers
Prev
2004;13(8):1392-8.
36. 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)
2007;55(7):1093-6.
37. Yokosuka A, Mimaki Y. Steroidal glycosides from the underground
parts of Trillium erectum and their cytotoxic activity. Phytochemistry
2008;69(15):2724-30.
38. 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
391
Asian J Pharm Clin Res, Vol 9, Issue 4, 2016, 386-392
Sharma et al.
underground part of Trillium tschonoskii. Chem Pharm Bull (Tokyo)
2007;55(4):679-81.
39. 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.
40. 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.
Statistics
429 Views | 4671 Downloads
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://innovareacademics.in/journals/index.php/ajpcr/article/view/12588.
Section
Original Article(s)