Simultaneous Estimation of Eugenol and Scopoletin from the in House <i>Avipattikar Churna</i> by RP-HPLC Method and Estimation of Scopoletin from Different Extracts of <i>Jalap</i>

Abstract

Background: Although standardising polyherbal medicine requires immediate attention, it is a tedious undertaking. Phytochemical profiling is a particularly useful tool for assessing the quality and effectiveness of polyherbal medicines, among various methods used for standardization. The proposal aimed to develop a precise RP-HPLC method for simultaneous estimation of eugenol and scopoletin in in-house Avipattikar churna. This method was also used to estimate scopoletin in various extracts of Ipomoea turpethum. Methods: The phytomarkers in Avipattikar churna, hydroalcoholic and alcoholic extracts of Jalap were estimated by RP-HPLC system. In this setup, RP-ODS C8 column was employed with methanol: water (30:70 v/v, 0.1% formic acid) at 1 ml/min for 0-10 minutes, and then with methanol: water (60:40 v/v) at 0.8 ml/min for 10.01-25 minutes. Detection was done at 280 nm for eugenol and 366 nm for scopoletin using a UV/VIS detector. The method was validated by performing validation parameters as per ICH guidelines. Results: The linearity of eugenol and scopoletin was performed, with correlation coefficients of 0.999 and 0.9969 respectively. In repeatability, % RSD was observed as 0.856 and 0.909 for eugenol and scopoletin correspondingly. The LOD (detection limit) of eugenol was 0.67 μg/mL and of scopoletin was 1.39 μg/mL. While LOQ (quantification limit) of eugenol was found as 2.04 μg/mL and 4.03 μg/mL for scopoletin. The % recovery was ranging from 102.96 - 100.45 % for eugenol and from 102.65 - 101.3 %w/w for scopoletin, after adding a pre-quantified amount (20 μg/mL) in the different concentrations of the standards. The eugenol and scopoletin were estimated 0.1366 %w/w and 0.0465 %w/w respectively in Avipattikar churna. The hydro alcoholic extract of Jalap showed presence of more scopoletin than in the alcoholic extract. Conclusion: The validated process is established as accurate, consistent and precise which results as a better standardization drive for Ayurvedic dosage forms.