Comparison between high performance liquid chromatography and high performance thin layer chromatography determination of Diosgenin from fenugreek seeds

Dheeraj H. Nagore, Pankaj S. Patil, Vinod V. Kuber


Background: Fenugreek is used in the treatment of many diseases as reported in Ayurveda. The chemical like Diosgenin is one of the important chemicals of fenugreek. Aim: To develop simple, rapid, accurate methods for estimation of Diosgenin from
fenugreek using high performance thin layer chromatography (HPTLC) and high performance liquid chromatography (HPLC).
Materials and Methods: The two chromatographic methods, HPTLC and HPLC, were developed and compared to quantitative
determination of Diosgenin in fenugreek seed extract. For both the analyses, the scanning wavelength was set at 205 nm. The Diosgenin content of seven different extracts collected from different locations was compared by HPLC and HPTLC. Statistical Analysis: The relative standard deviation and correlation coefficient (r2) were calculated by standard Formulae. Results: The comparison was done in HPTLC and HPLC results and all the results are found to be within the limits and showed no significant difference. Conclusion: Both the methods were accurate and specific for the analysis of Diosgenin.
Key words: Diosgenin, fenugreek, high performance liquid chromatography, high performance thin layer chromatography,

Full Text:



Bin‑Hafeez B, Haque R, Parvez S, Pandey S, Sayeed I,

Raisuddin S. Immunomodulatory effects of fenugreek (Trigonella

foenum graecum L.) extract in mice. Int Immunopharmacol


Gopu CL, Gilda SS, Paradkar AR, Mahadik KR. Development and

validation of a densitometric tlc method for analysis of trigonelline

and 4‑hydroxyisoleucine in fenugreek seeds. Acta Chromatogr


Srinivasan K. Fenugreek (Trigonella foenum‑graecum): A review

of health beneficial physiological effects. Food Rev Int


Onweluzo JC, Obanu ZA, Onuha KC. Functional properties of

some lesser tropical legumes. J Food Sci Tech 1994;31:302‑06.

Acharya S, Srichamroen A, Basu S, Ooraikul B, Basu T. Improvement

in the nutraceutical properties of fenugreek (Trigonella

foenum‑graecum L.). J Sci Technol 2006;28:1‑9.

Chatterjee S, Prasad SV, Sharma A. Bioactive lipid constituents of

fenugreek. Food Chem 2010;119:349‑53.

Shishodia S, Aggarwal BB. Diosgenin inhibits osteoclastogenesis,

invasion, and proliferation through the downregulation of Akt,

I kappa B kinase activation and NF‑kappa B‑regulated gene

expression. Oncogene 2006;25:1463‑73.

Liagre B, Vergne‑Salle P, Corbiere C, Charissoux JL, Beneytout JL.

Diosgenin, a plant steroid, induces apoptosis in human rheumatoid

arthritis synoviocytes with cyclooxygenase‑2 overexpression.

Arthritis Res Ther 2004;6:R373‑83.

Mehrafarin A, Qaderi A, Rezazadeh S, Naghdi BH,

How to cite this article: Nagore DH, Patil PS, Kuber VV. Comparison

between high performance liquid chromatography and high performance thin

layer chromatography determination of Diosgenin from fenugreek seeds. Int

J Green Pharm 2012;6:315-20.

Source of Support: Nil, Conflict of Interest: None declared.

Noormohammadi G. Bioengineering of important secondary

metabolites and metabolic pathways in fenugreek (Trigonella

foenum‑graecum L.). J Med Plants 2010;9:1‑18.

Anonymous. The Ayurvedic Pharmacopoeia of India. New Delhi:

Controller of publications, Government of India; 1999.

p. 112‑3.

Cardellina JH 2nd. Challenges and opportunities confronting

the botanical dietary supplement industry. J Nat Prod


ICH. International Conference on Harmonization Q2B: Validation

of Analytical Procedures. US FDA Fed Reg; 1997. p. 463‑7.

Chopra S, Ahmad FJ, Khar RK, Motwani SK, Mahdi S, Iqbal Z,

et al. Validated high‑performance thin‑layer chromatography

method for determination of trigonelline in herbal extract and

pharmaceutical dosage form. Anal Chim Acta 2006;577:46‑51.



  • There are currently no refbacks.