Optimizing the method for quantification of apigenin and quercetin in the thai herbal Sattakavata formula by ultra-performance liquid chromatography coupled with mass spectrometer

Abstract

© 2020, Faculty of Pharmaceutical Sciences, Chulalongkorn University. All rights reserved. Introduction: Sattakavata formula (SVF) is a Thai traditional medicine from Ayurved Siriraj for a relief of joint pain. It comprises six ingredients, namely, Andrographis paniculata (Burm. F.) Wall. ex Nees (AP), Cinnamomum sp. (CN), Morus alba Linn. (MA), Crateva adansonii DC. subsp. trifoliata (Roxb.) Jacobs (CA), Moringa oleifera Lam. (MO), and Acorus calamus Linn. (AC). There was no method for identification and quantification of SVF before. Objective: The objective of the study was to establish a method for identification and quantification of apigenin and quercetin in SVF and its components using ultra-performance liquid chromatography–mass spectrometry (UPLC-MS). Materials and Methods: SVF and six components were extracted with 80% ethanol by ultrasonication. The separation was performed in an RP18 column using a gradient elution with 0.1% formic acid in deionized water and acetonitrile. The method was fully validated in terms of precision, limit of detection, limit of quantification, and recovery. Results: Apigenin was found in SVF, AP, and CN, and quercetin was found in SVF, CN, and MA. There was a good linearity (R2 > 0.999) in the range of 100–1400 ng/ml for apigenin and 300–3000 ng/ml for quercetin. The recovery of apigenin and quercetin was in the range of 88.33–111.52% and 90.71–109.17%, respectively. Relative standard deviations of precision in apigenin and quercetin were 0.62–6.02 and 0.65–4.01, respectively. Conclusion: A reliable UPLC-MS method for identification and quantification of apigenin and quercetin in SVF was successfully established in this study. The method is useful in the quality control of the herbal medicines and can be used routinely.