Analysis of Phenolics in <I>Clematis gouriana</I> Roxb. Ex Dc. Aerial Parts by HPLC and Method Validation

Abstract

Fast and economical analytical methods are required for quality control of herbal medicines/ extracts. So, a simple and accurate RP-HPLC method with photodiode array detection was developed for the identification and quantification of phytochemicals in Clematis gouriana Roxb. ex DC. aerial parts. After several attempts the best HPLC method was developed on Waters C<sub>18</sub> analytical column with column temperature of 30 °C, using a gradient elution solvent consisting of water (0.1% formic acid): acetonitrile at 1mL/min flow rate; and detection was done at 254 and 330 nm. Four phenolic compounds quantified were p-coumaric acid, hyperoside, methyl caffeate, and caffeic acid. Caffeic acid was detected as the major phytoconstituent that may be considered as a marker compound for C. gouriana. Content of the four phenolic compounds extracted by reflux, ultra-sonication, and microwave-assisted extraction was also compared. The developed HPLC method showed good linearity (r<sup>18</sup> >0.998), high precision (<2%) and good recovery (98.8-102.7%) for the studied four compounds, and validation parameters were also found within permissible limits as per the International Council of Harmonization guideline Q2 (R1). To the best of our knowledge, this is the first report on the quantification of phytochemicals and validation of the HPLC method for Clematis gouriana aerial parts.