Optimization and validation of reverse phase HPLC and HPTLC method for simultaneous quantification of vasicine and vasicinone in Sida Species

Quantification of bioactive principles through modern analytical tools is essential for establishing the authenticity and creditability of prescription and usage of herbal drugs. In the present study, simultaneous quantification of vasicine and vasicinone, present in Sida species (Malvaceae), by reverse phase HPLC and HPTLC methods were developed. In the RP-HPLC method, the extracts were resolved using a mobile phase of acetonitrile - 0.1 M phosphate buffer - glacial acetic acid (15:85:1, v/v/v) with pH 4.0 on a C18-ODS-Hypersil column in isocratic mode. The retention times of vasicine and vasicinone were 5 and 8.7 min, respectively. In the HPTLC method, mobile phase of ethyl acetate, methanol, ammonia (8:2: 0.2, v/v) was used on precoated plate of silica gel 60 F254 and quantified by densitometric method. Validation of the methods was done to demonstrate its selectivity, linearity, precision and accuracy as recommended in the ICH guidelines. Excellent linear behaviors over the investigated concentration ranges were observed with the values of R2 higher than 0.998 for both the analytes. Recovery values between 97 - 101%, and correlation coefficient between (linear dynamic range) 0.977 - 0.9999 shows that the developed methods were accurate and precise. These methods can be employed for the routine analysis of the quality of herbal extracts and formulations.             Key words: Sida cordifolia, Sida acuta, Malvaceae, quantification, validation, HPLC, HPTLC.