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  • Title: Column-switching high-performance liquid chromatographic assay for determination of apigenin and acacetin in human urine with ultraviolet absorbance detection.
    Author: Nielsen SE, Dragsted LO.
    Journal: J Chromatogr B Biomed Sci Appl; 1998 Aug 25; 713(2):379-86. PubMed ID: 9746253.
    Abstract:
    A high-performance liquid chromatographic (HPLC) method is described for the determination of apigenin and the 4'-methylated derivative acacetin in human urine using column-switching and ultraviolet (UV) absorbance detection. Urine samples were enzymatically hydrolysed and solid-phase extracted prior to injection onto the HPLC system. Prior to elution of apigenin and the internal standard, 5,7,8-trihydroxyflavone, from the first column used for sample clean-up, the six-port valve was switched to the second column for analysis with UV detection. Detection of apigenin was precise and reproducible, with a limit of quantification of 10 ng ml(-1) urine. Detection and quantification of acacetin was linear down to 70 ng ml(-1) urine. The method has been successfully applied to determine the level of apigenin in 100 human urine samples from an intervention study with parsley.
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