Imoto et al. (1991) reported that selleck products GA content was confirmed through HPLC in methanol leaf extracts of G. sylvestre [34]. Figure 5(b) describes the GA content of in intact leaf explants (19.52mg/gd.w), which was increased compared to Figure 5(c) of in vitro callus culture of MS medium supplemented with OPGRS (12.22mg/gd.w). Many authors had isolated and identified GA earlier in leaf explants of methanol extracts. In 1989, Yoshikawa and coworkers isolated GAs from a hot water extract of G. sylvestre, which they named GA I, II, III, IV, V, VI, and VII, respectively, and evaluated using HPLC [35, 36]. For GA enhancement, OPGRs culture was kept under physical-chemical stress conditions determined by growth curve analysis. Blue light with OPGRs induced the maximum GA (53.94mg/gd.

w) (Figure 5(d)) rather than 5% sucrose treatment (33.39mg/gd.w). However, with other physical stress conditions, the GA was reduced in this order 12h photoperiod (26.27mg/gd.w), red light (8.90mg/gd.w), green light (5.72mg/gd.w), and 30��C (2.9mg/gd.w). In case of dark treatment, GA content was absent. We have reported, that in vitro callus of G. sylvestre significantly increased the pancreatic ��-cells and maintained the body weight, pancreas weight, liver weight and liver glycogen level in alloxan-induced diabetic Wistar rats [13].Figure 5(a) Standard gymnemic acid; (b) in vivo leaf; (c) OPGRs MS + 2,4-D (1.5 mg/L) with KN (0.5 mg/L); (d) blue light treatment with OPGRs.3.8. Linearity, Precision, Recovery, and Robustness in HPLCAs per the ICH guidelines, the method validation parameters checked were linearity, accuracy, precision, limit of detection, limit of quantification, and robustness.

The linearity of the method was determined Carfilzomib at three concentrations (10�C30ug/mL) of GA. 20ug/mL GA results show that an excellent correlation exists between response factor and concentration of GA within the concentration range indicated above.The accuracy of the method was determined by recovery experiments. The recovery studies were carried out at three levels of 80, 100, and 120%, and the percentage recovery was calculated. Our studies recovery was within the range of 100 �� 2% which indicates accuracy of the method. The precision of the method was demonstrated by interday and intraday variation studies. In the intraday studies, 3 repeated injections of standard and sample solution were made in a day and the response factor of GA peaks and percentage were calculated. In the interday variation studies, 3 repeated injections of standard and sample solutions were made on 3 consecutive days, and response factor of GA peak and percentage were measured.