In this study, a sensitive, selective and reproducible liquid chromatography-tandem mass spectrometry method for the simultaneous determination of 1,5-dicaffeoylquinic acid (1,5-DCQA) and its active metabolites, 1-caffeoyl-5-feruoylquinic acid and 1,5-O-diferuoylquinic acid, in human plasma, using puerarin as internal standard, was developed and validated. Analytes were extracted from plasma samples by liquid-liquid extraction with ethyl acetate, separated on a C(18) reversed-phase column with water containing 5 mM ammonium acetate and acetonitrile as the mobile phase and detected by electrospray ionization mass spectrometry in negative selected reaction monitoring mode. The accuracy and precision of the method were acceptable and linearity was good over the range 1-200 ng/mL for each analyte. In addition, the selectivity, extraction recovery and matrix effect were satisfactory too. The validated LC-MS/MS method was successfully applied to phase II clinical pharmacokinetic study of 1,5-DCQA in patients.
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