Validation of an RP-HPLC method for quantitative analysis of phikud navakot extract using the standard addition method

Abstract

The aim of the study was to develop a reversed phase-high performance liquid chromatography (RP-HPLC) analytical method for monitoring three analytical markers, gallic acid, vanillic acid and ferulic acid, in Phikud Navakot extract (NVK-E), using the standard addition method. RP-HPLC was performed under gradient conditions of methanol and 1% v/v acetic acid (pH 2.7) at a flow rate of 1.0 ml/min with a total run time of 70 min. A C18 (4.6 x 250 mm, 5 µm) HPLC column was used. The column oven was controlled at 27ºC and DAD was adjusted from 190-400 nm with a monitoring wavelength of 270 nm. The results for all three analytical markers showed good linearity with correlation coefficients of >0.99. The percentage recovery of each analytical marker was between 92-105%. The precision of the system, presented in terms of HorRatr values, were 2 for all three markers. The linearity, accuracy and precision of the system complied with AOAC Guidelines for Single Laboratory Validation of Chemical Methods for Dietary Supplements and Botanicals (2013). These results indicated that the developed RP-HPLC analytical method can be utilized for monitoring the quality of NVK-E.