Purification and HPLC Analysis of Caffeoylquinic Acids from Kudingcha Made from Ilex kudingcha C.J. Tseng

Abstract

Six caffeoylquinic acid(CQA) derivatives were separated by column chromatography and semi-preparative high performance liquid chromatography(HPLC) from the crude extract of Ilex kudingcha C.J.Tseng.An HPLC method with DAD and TSKgel ODS-80 TsQA column was developed for the simultaneous determination of CQAs in Kudingcha made from Ilex kudingcha C.J.Tseng.The results demonstrated that six CQA derivatives with 95% purity,3-CQA,4-CQA,5-CQA,3,4-diCQA,3,5-diCQA and 4,5-diCQA,could be obtained through the purification of Ilex kudingcha C.J.Tseng by semi-preparative high performance liquid chromatography(HPLC).The chromatographic conditions were determined as follows:oven temperature set at 40 ℃,gradient elution with methanol/formic acid(5‰) with a flow rate 0.5 mL/min,and detection at a wavelength of 326 nm.As results,good linearity was observed with correlation coefficients of 0.9992–0.9999,the average recovery of the CQA derivatives from spiked samples was 96.34%,with relative standard deviations(RSD) less than 3%(n = 3).