Simultaneous Quantification of Pharmacologically Active Alkaloid Metabolites Colchicine and Gloriosine in Gloriosa Superba L. collected from Western Ghats (India) and Adjoining Areas for the Identification of Elite Chemotype(s).

Abstract

Gloriosa superba is a valuable Ayurvedic medicinal plant and is in high demand in the world market for its colchicine content, which is used to treat gout. The study aims (1) to record the metabolic variations in major bioactive metabolites, colchicine and gloriosine, in the natural populations of G. superba from Western Ghats and adjoining areas in India and (2) to develop HPTLC protocol for the identification of elite chemotypes of species and regulation of quality raw material, extract, and finished material. Simultaneous quantification of colchicine and gloriosine in 22 natural populations through validated HPTLC as per ICH guidelines. Colchicine and gloriosine were identified at Rf 0.51 ± 0.03 and 0.41 ± 0.05 and the content varied from 0.021 to 0.86% and 0.003 to 0.198%. The method was found linear at a concentration range of 0.1-0.7 µg/spot, and LOD (3.3 σ/S) and LOQ (10 σ/S) was 0.71 and 2.16 µg/spot. The method was precise in the concentration range of 100-300 ng/spot, with 98.29% and 101.12% recovery (% RSD) for colchicine and gloriosine. Subsequently, four elite chemotypes were identified based on cluster analysis of metabolite content. The developed HPTLC method is linear, accurate, precise, and robust for simultaneous quantification of colchicine and gloriosine metabolite(s). Intraspecific metabolic variation was significant among the collected population, leading to the identification of four elite chemotypes. Colchicine is an industrially viable metabolite and is therefore quintessential to the development of an economical and analytical method to regulate the quality of raw material, extract, and finished products.