In Vitro Production and Efficient Quantification of Major Phytopharmaceuticals in an Endangered Medicinal Herb, Swertiachirata

Abstract

Swertiachirata is an endangered medicinal herb with a wide range of pharmacological activities, particularly as an anti-diabetic in an herbal formulation Diabecon (Himalaya®). The endangered status has resulted in short supply of plant material, thereby, resulting in adulteration practices. It is, therefore, of paramount importance to develop a shoot biomass production system enriched with major chemical constituents, swertiamarin, mangiferin and amarogentin and their rapid quantification. In vitro shoot multiplication occurred within 22-25 days on MS medium supplemented with KN (2mg/L) + GA3 (3mg/L) + IBA (2mg/L) + sucrose 3% (w/v). Swertiamarin content was highest (9.33μg/mg) in nutrient medium containing IBA (2mg/L) and GA3 (3mg/L) at 15±1°C after 30 days of inoculation. Mangiferin and amarogentin contents were highest, 2.99 μg/mg and 1.03 μg/mg, respectively in cultures grown in nutrient media containing IBA(2mg/L) and KN (2mg/L) at 25±1°C in 30 days. We have optimized a highly sensitive and efficient HPLC method for the quantification of these medicinal compounds simultaneously for quality control and plant material standardization. The retention for peak due to swertiamarin, mangiferin and amarogentin was about 8.5 min, 13.6 min and 21.7 min, respectively. The quantification method is based on rapid ultra-sonic wave assisted extraction with sharp peaks for high limit of detection(LOD) and high limit of quantification(LOQ) simultaneously in 45 minutes of time duration contrary to previous methods