HPLC Method of Analysis for Determination and Standardization of Luteolin and Vanillic Acid in Dry Extract of Paronychia Argentea Lam.

Abstract

The main objective of this study was to establish a chromatographic method for analysis, determination and standardization of the two main components vanillic acid and luteolin as major components in Paronychia argentea Lam dry extract. This analytical method was designed to be a simple and fast with an appropriate separation of the two main components of the extract. High pressure liquid chromatography (HPLC) method of analysis was developed to quantitatively determine, identify and standardize the two main active constituents in the pharmaceutical dry extract against luteolin and vanillic acid as primary reference standards as it is the major active constituents of the dry extract of P. Argentea, where the linearity obtained was higher than R2 = 0.99981 and 0.99908 respectively. Although the method was proven to be suitable, further specific analysis validation was conducted to include the following: linearity, precision, range, limit of detection, limit of quantitation and filter compatibility. The luteolin and vanillic acid were completely separated from the other components in the herbal dry extract with an Rf value of 1.3 and 5.7 minutes respectively. The concentration of Luteolin is 0.4% while vanillic acid content is 0.1% in the dry extract.