Development and validation of a robust high-performance thin layer chromatographic (HPTLC) method for the analysis of Lovastatin in Higher Basidiomycetous mushrooms

Abstract

Hypercholesteromia is one of the main risk factors of cardiovascular disease, causing high blood pressure and atherosclerosis. This has necessitated development of methods or substances which would reduce cholesterol uptake from the digestive tract or destroy cholesterol by enzymatic reactions. One of the practical approaches to protect the human body from high cholesterol levels is to inhibit the cholesterol-synthesizing ability of the organism. Lovastatin is a metabolite of mold fungi from the different genera. However, its presence has also been detected in fruiting bodies and submerged cultivated mycelia of some species of Higher Basidiomycetous fungus, especially in the species of genus Pleurotus – P. ostreatus and P. eryngii. We are screening Higher Basidiomycetous mushrooms for identification of commercially recoverable quantities of lovastatin.  Since lovastatin quantification in a large number of fungal samples is required, a simple, rapid and accurate analytical method was required. Use of HPTLC for quantitative measurements of lovastatin in fungal biomass has not yet been reported. In the present study a simple, precise, specific and accurate TLC Densitometric method for estimation of lovastatin in Higher Basidiomycetous mushrooms was developed, validated and used for estimation of lovastatin content in the cultured mycelia of Pleurotus spp. The develop method was validated for linearity, accuracy, precision, LOD, LOQ, spiking, robustness as per the ICH guidelines. The R_f and % recovery value for Lovastatin 0.52 and 99.15 respectively. Developed analytical methods showed good separation and recovery of Lovastatin.