Development of a HPTLC method with iterative calibration approach for quantitative evaluation of kinsenoside content in different types of Anoectochilus roxburghii

Abstract

Anoectochilus roxburghii (Wall.) Lindl. (Orchidaceae) or more commonly known as Jin Xian Lian is a widely adapted medicinal herb in the Asian continent. However, till date there are limited rapid analytical method such as thin layer chromatography method that acts as standard for quantifying the active ingredients of A. roxburghii. In this research, a simple and rapid method high performance thin layer chromatography (HPTLC)-anisaldehyde-sulfuric acid derivative with iterative calibration method was developed and validated to quantify the non UV sensitive bioactive compound kinsenoside in wild, cultivated and tissue cultured A. roxburghii plants. The optimum mobile phase identified for kinsenoside in A. roxburghii was chloroform/methanol/water (45:30:4,%v/v/v). It was also proved that HPTLC method for determination of kinsenoside showed good accuracy, precision, stability and repeatability, with wide working concentration range of 0.099–6.365 mg/mL. Intra-day and inter-day precision (% RSD) of the developed HPTLC method were 5.08–7.57% and 6.72–8.72%, respectively. The % RSD of iterative calibration method was 3.139%. Through this study, it can be concluded that the proposed HPTLC method with iterative calibration approach was successfully applied for quantitative analysis of kinsenoside in three types of A. roxburghii plants.