Method Development for Quantitative Determination of Diosgenin from the Seeds of Fenugreek, <i>Trigonella foenum-graecum</i> L.

Abstract

Introduction . Modern pharmacognostic research is aimed at searching for plant biologically active individual compounds (RBAIS) isolated from plant extracts. Aim . Study of the content of the steroid sapogenin diosgenin in fenugreek seeds in plant extracts by HPLC-UV method. Materials and methods . The object of study was raw materials-fenugreek seeds produced as medicinal plant raw materials by LLC «Sage» (Irkutsk). Series of extraction and chromatographic separation by HPLC-UV method were performed. Results and discussion . In this work, the method of diosgenin extraction was successfully optimized (after acid hydrolysis of dioscin with a 5 % aqueous solution of hydrochloric acid when heated for at least 4 hours in a sand bath at a power of 150 W) from raw fenugreek seeds using a 50 % aqueous solution of isopropanol, h.h. A simplified technique for chromatographic separation of diosgenin from fenugreek seed extracts using an isocratic mode of 99.9 % acetonitrile elution (h.h.) was proposed. Conclusion . The average content of diosgenin (after acid hydrolysis of dioscin) in fenugreek seeds, taking into account the humidity of raw materials, is (M ± σ ) 5,3 ± 0,05 mg/g (95 % CI: 5,1-5,4 mg/g; n = 6).