Biomarkers identification of Lycopodiaceae and Huperziaceae species from peninsular Malaysia with HPLC chromatographic profiling and partial least square analysis

Abstract

AbstractCrude herbal extracts are complex matrix consisting large quantity of plant metabolites. The identification of plant metabolite exhibiting bioactivity using the conventional bioactivity‐guided isolation process is a tedious and time‐consuming method. Club mosses from the Lycopodiceace and Huperziaceae families from Peninsular Malaysia is used as a model for this study since the alkaloid, (‐)‐huperzine A, exhibited acetylcholinesterase inhibition and used for the treatment of Alzheimer disease. The aim of this study is to evaluate the feasibility of using high‐performance liquid chromatographic profile and applying partial least square projection to identify the biomarkers. A high‐performance liquid chromatography gradient system with increasing amount of acetonitrile in deionized water and an ODS‐3 column (250 × 4.6 mm) was developed for the acquisition of the chromatographic profiles. A colorimetric assay was used to analyze the acetylcholinesterase inhibition activity. Both the pretreated high‐performance liquid chromatography integration data and acetylcholinesterase inhibition results were subjected to chemometric data analysis. From the unsupervised principal component analysis results, no outliers were detected and the data was further subjected to partial least square projections. Two biomarkers were identified and were isolated and structurally elucidated as 6β‐hydroxyhuperzine A and (‐)‐huperzine A.