TLC densitometric quantification of picrosides (picroside-I and picroside-II) in Picrorhiza kurroa and its substitute Picrorhiza scrophulariiflora and their antioxidant studies.

Picroside-I and picroside-II are known bioactive metabolites in Picrorhiza species. In the present study a simple, precise method has been established for the simultaneous determination of picrosides (picroside-I and picroside-II) in two different Picrorhiza species, P. kurroa and P. scrophulariiflora. This method was also validated for accuracy, precision, robustness, limit of detection and quantification, repeatability and recovery, according to International Conference of Harmonization guidelines. Separation and quantification was achieved by HPTLC using as the mobile phase chloroform-methanol (88:12, v/v) on precoated silica gel 60F(254) aluminum plates. Densitometric determination was carried out at wavelength λ(max) 254 nm in UV absorbance mode. Comparative study also revealed that picroside-I and picroside-II are higher in P. scrophulariiflora than P. kurroa. Picroside-I content was found to be 1.258 and 1.611%, and picroside-II was estimated as 0.481 and 0.613% in P. kurroa and P. scrophulariiflora, respectively. Antioxidant potential of these two Picrorhiza species was also studied using DPPH. At a concentration of 0.1 mg/mL the scavenging activities of P. kurroa and P. scrophulariiflora were found to 37.70 and 34.30%, respectively.