Development and Validation of HPLC And HPTLC for Simultaneous Analysis of E and Z Guggulsterone, A-11–KBA And 11–KBA from Herbal Formulation

Abstract

RP-HPLC and HPTLC are two easy, sensitive, efficient, and accurate procedures that allow for simultaneous estimates of Z and E guggulsterone, A-11-KBA, and 11-KBA. For the HPLC method, we recommend the use of a symmetry C18 column. Using a solvent gradient based on solvent A (orthophosphoric acid) and solvent B (methanol), the effluent was monitored at 250 nm with a 1.0 mL/min flow rate. The peaks of 11-KBA and A-11-KBA were eluted at 5.8 and 6.3 minutes, while those of Z and E-guggulsterone were at 4.8 and 5.3 minutes. For the HPTLC method of separation, a silica gel layer was applied on an aluminum plate prewashed in methanol using a Camag Linomat V applicator fitted with a 100 μL syringe. The linear expansion was carried out using a solvent mixture of n-hexane, chloroform, ethyl acetate, and methanol (v/v/v: 10:3:3:1, respectively). Camag T.L.C. scanner III (V 1.4.3.6336), operating in reflectance-absorbance mode at 254 nm and controlled by win CATS software, was used to carry out the densitometric scanning. The R.F. resolutions for 11-KBA, A-11-KBA, E-guggulsterone, and Z-guggulsterone in the selected mobile phase were 0.68, 0.61, 0.39, and 0.28, respectively. The linearity, accuracy, and precision of the techniques were all confirmed. The proposed methods were successful in estimating E- and Z-guggulsterone as well as 11-KBA and A-11-KBA.