Method development and validation for quantification of Yohimbine in root of Rauwolfia serpentina by HPTLC

Abstract

Plants are a priceless and vital resource that give us medicine, food, fibre, and all other necessities. The evergreen shrub, Rauwolfia serpentina (L.) Benth. ex Kurz., also known as Indian snakeroot, sarpagandha is a member of the Apocynaceae family. Rauwolfia serpentina has fewer negative effects and an abundance of phytochemicals in the roots and leaves of the plant, it shows a wide range of therapeutic activity. With the goal of developing a densitometric high-performance thin-layer chromatography (HPTLC) method, in order to quantify Yohimbine hydrochloride in Rauwolfia roots, it will help to insure the collection and harvesting time. The five distinct brands of market Rauwolfia root samples are compared with in-house samples prepared in the laboratory. The silica gel 60 F254 high-performance thin-layer chromatography plates were used with toluene, Toluene : Ethyl acetate : Diethylamine (7:1.5:1.5 v/v/v) as a mobile phase. The validation parameters for the developed method were carried out as per the International Conference on Harmonisation (ICH) guideline Q2 (R1). The calibration curve was found to be linear over a range of 1-7 μg/spot for yohimbine. The method involves densitometric detection at 280 nm. The relative standard deviations (RSD) for precision were found to be lower than 2% for both analytes. Percent recoveries for accuracy were performed and found to be 98.5-99.98% for yohimbine, The LOD and LOQ for Yohimbine were 0.08 μg/ml and 0.24 μg/ml, respectively. Validation parameters were carried out as per International Conference on Harmonisation (ICH) guideline Q2 (R1). The Rf value of 0.64 have been observed for Yohimbine. This HPTLC densitometric method is highly suitable for the analysis of Yohimbine present in the dosage of marketed herbal products of sarpagandha root without any interference.