Simultaneous estimation of three triterpenoids from Carissa carandas using validated high performance liquid chromatography

Abstract

Background: Carissa carandas L. (Apocynaceae), an important fruit of tropical and subtropical arid region, has a great potential for the development of a new phytotherapeutic medicine. Due to an increase in commercial products of this fruit in the global market, an extensive phytochemical comparison seems imperative to understand its internal quality. Aims and Objective: To develop and validate an HPLC-DAD method for simultaneous estimation of three triterpenoids from C. carandas. Materials and Methods: A simple, sensitive and rapid HPLC-DAD method was developed for simultaneous estimation of lupeol, stigmasterol, and b-sitosterol from C. carandas fruits. Isocratic elution at a flow rate of 1.0 mL/min was employed on C18 Cosmosil column with PDA detection at 210 nm using acetonitrile and ethanol. Different parts of C. carandas and its fruits collected from different geographical region were screened for lupeol, stigmasterol, and b-sitosterol contents. Results: Proposed method for simultaneous estimation of lupeol, stigmasterol and b-sitosterol was found highly specific, reliable, and sensitive which allowed good recovery of all analytes. It was validated as per ICH guidelines and proved to have good linearity, precision, repeatability, and stability. It was efficiently applied to screen different samples of C. carandas providing a mean to assess the quality and authenticity. Conclusion: The HPLC method could be an effective technique for routine analysis of lupeol, stigmasterol, and b-sitosterol simultaneously and monitoring the quality of fruit and its available products in cosmetic and pharmaceutical companies. Key words: Carissa carandas, simultaneous estimation, triterpenoids, validation