A Rapid, Validated HPLC Analytical Method for Simultaneous Quantitation of Phytoconstituents in Asteracantha Longifolia (L.) Nees

Abstract: Asteracantha longifolia (L.) Nees a small, spiny weeds found in marshy places. It has effective antitumor, hypoglycemic, aphrodisiac, antibacterial, free radical scavenging, lipid peroxidation, and hepatoprotective properties1. Asteracantha longifolia (L.) Nee is an essential herb in many traditional and herbal medicines. Active phytoconstituents like Betulin, Lupeol, Stigmasterol, and β-sitosterol are reported to accord the herb importance as a medicinal plant in modern and traditional medicine. The plant's usefulness in many traditional medicines has made it crucial to quantify the essential phytoconstituents. However, prior art analytical methods profoundly lack rapidness and simultaneous quantitation of different phytoconstituents. This research work has validated the analytical method for simultaneous quantitation of Betulin, Lupeol, Stigmasterol, and β-sitosterol with HPLC- PDA, in line with the guidelines by ICH. The research deals with a systematic approach to developing, optimizing, and validating a chromatographic method. The optimized chromatographic conditions resulted in the best separation for four phytoconstituents. Furthermore, method validation parameters like Specificity, LOD (0.07µg/mL) and LOQ (0.2µg/mL), Linearity, Precision, Intermediate Precision, Recovery, and Range are found to be within limits laid by ICH.