HPTLC method development and validation of stigmasterol from different extracts of Tagetes erecta and Capsicum annuum
A new, simple, sensitive, selective, precise and robust high-performance thin-layer chromatographic (HPTLC) method for analysis of stigmasterol was developed and validated for the determination of stigmasterol in different extracts. A new, simple, sensitive, selective, precise and robust high-performance thin-layer chromatographic (HPTLC) method for analysis of stigmasterol was developed and validated for the determination of stigmasterol in different extracts. Analysis of stigmasterol was performed on TLC aluminium plates pre-coated with silica gel 60F-254 as the stationary phase. Linear ascending development was carried out in twin trough glass chamber saturated with mobile phase consisting of Hexane : Acetone (8:2 v/v) at room temperature. (25 °C ± 2 °C) Camag TLC scanner III was used for spectrodensitometric scanning and analysis in absorbance mode at 490nm. The system was found to give compact spots for stigmasterol. (Rf value of 0.44 ± 0.02) The linear regression analysis data for the calibration plots showed good linear relationship with r2=0.9997 ± 0.0002 in the concentration range 200-1200 ng spot−1 with respect to peak area. According to the International Conference on Harmonization (ICH) guidelines the method was validated for precision, recovery and robustness. Statistical analysis of the data showed that the method is reproducible and selective for the estimation of stigmasterol.
Keywords: Tagetes erecta, Capsicum annuum, stigmasterol, HPTLC, method validation
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