Development of a validated HPTLC method for quantification of stigmasterol in marine algae

This study presents the report of HPTLC method which has been developed and validated for quantification of stigmasterol in marine algae. The separation was performed on HPTLC Silica gel 60 F254 glass plate 20 cm x 10 cm. Good separation was performed in mobile phase using n-hexane: ethyl acetate: acetic acid (60: 36: 4, v/v/v). Determination and quantitation were achieved by densitometric scanning at 550 nm in absorbance mode. This method gave samples spot at Rr = 0.78 corresponding to stigmasterol standard. The method was validated in terms of precision and repeatability. Linearity range for stigmasterol was 0.5-10 µg/spot with correlation coefficient, R2=0.9746. The LOD and LOQ were found to be 0.61 µg and 2.04 µg respectively. The highest amount of stigmasterol content obtained from marine algae sample (track 6 and 16) were 1.90 % w/w and 1.94 % w/w respectively. HPTLC method was found to be a simple, precise, accurate and convenient method for rapid screening of active constituents present in ethanol and ethyl acetate extracts. It can be used for analysis and routine quality control of herbal materials and formulations containing marine algae.