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

Sattakavata formula (SVF) is a Thai traditional medicine from Ayurved Siriraj for a relief of joint pain. It comprises six ingredients, namely Andrographis paniculata (AP), Cinnamomum sp. (CN), Morus alba (MA), Crateva adansonii DC. subsp. trifoliata (CA), Moringa oleifera (MO) and Acorus calamus (AC). There was no method for identification and quantification of SVF before. Therefore, this paper establishes a method for identification and quantification of Apigenin and Quercetin in SVF and its components using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS). SVF and six components were extracted with 80% ethanol. 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. Apigenin was found in SVF, AP, CN, and Quercetin was found in SVF, CN and MA. There was a good linearity (R2 > 0.999) in the range of 100 -1,400 ng/ml for Apigenin and 300–3,000 ng/ml for Quercetin. The recovery of Apigenin and Quercetin were in the range of 88.33-111.52% and 90.71-109.17%, respectively. RSDs of precision in apigenin and quercetin were 0.62-6.02 and 0.65-4.01, respectively. This method is useful in the quality control of the herbal medicines and can be used routinely.