Development and Validation of HPTLC Method for Simultaneous Estimation of Diosgenin and Quercetin in Fenugreek Seeds (Trigonella foenum-graceum)

Abstract

A sensitive, fast, and reproducible high performance thin-layer chromatographic method has been developed for simultaneous analysis of diosgenin and quercetin from fenugreek seeds, using TLC aluminium plates precoated with silica gel G60F254. Among the different combinations of mobile phases used, best separation was achieved in Toluene-ethyl acetate-formic acid (5 : 4 : 1, v/v/v). Densitometric scanning of the plates directly at 275nm was used for analysis of quercetin. While as for analysis of diosgenin, plates were scanned at 450nm after spraying with anisaldehyde-sulphuric acid reagent. The retardation factorvalue of diosgenin and quercetin was found to be 0.69 ± 0.02 and 0.57 ± 0.02, respectively. The method was validated for specificity, precision (intraday and interday), accuracy, and robustness. Accuracy of the method was checked by recovery study of three different levels with the average percentage recovery of 99.13 ± 0.26 for diosgenin and 99.63 ± 0.34 for quercetin, respectively. Dried fenugreek seed samples were found to contain diosgenin in the range of 0.113–0.135% (w/w) and quercetin in the range of 0.009–0.012% (w/w). The present method is being reported for the first time and can be used for routine quality control and quantification of these marker compounds in various plant samples, extracts, and market formulations.

Diosgenin: Recent Highlights on Pharmacology and Analytical Methodology

“…Both methods were precise and specific and there was no statistical significant difference between them. Other HPTLC methods have been validated for the determination of diosgenin also in fenugreek seeds and in marketed formulations [101, 102]. In addition, by means of the HPTLC technique, the diosgenin content in fifteen different Trigonella species was determined, including seeds and aerial parts of the plant, and it can be concluded that the seeds of the species Trigonella foenum graecum present the highest diosgenin level [103].…”

Characterization of extracted phenolics from black cumin ( Nigella sativa linn), coriander seed ( Coriandrum sativum L.), and fenugreek seed ( Trigonella foenum-graecum )

“…[56] Diosgenin in FS were estimated through HPLC and found 29.65 μg/ml and 5.087 mg/g, respectively. [57,58]…”

Chromatographic Fingerprinting and Quantitative Analysis of Marker in the Extract of Gloriosa superba Tubers Collected from Some Region of Chhattisgarh

“…In the accuracy study, the percent recoveries were between 98.6-99.7%, which indicated adequate reliability, repeatability, and accuracy of the method. 59 The percent RSD obtained for intraday and inter-day precision were in between 0.134-0.645, which is significant as per ICH guidelines and suggested that the method is precise. In robustness study, low present RSD was indicated that the proposed method is robust.…”