Column-switching high-performance liquid chromatographic assay for determination of apigenin and acacetin in human urine with ultraviolet absorbance detection.
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.
Nielsen
S E
SE
Institute of Foodsafety and Toxicology, Danish Veterinary and Food Administration, Søborg, Denmark.
Dragsted
L O
LO
D016588:Anticarcinogenic Agents / Q000652:urine*; D019661:Apiaceae / Q000737:chemistry; D001192:Arylsulfatases; D020843:Chamomile; D002851:Chromatography, High Pressure Liquid; D047309:Flavones; D005419:Flavonoids / Q000032:analysis* / Q000737:chemistry / Q000652:urine*; D005966:Glucuronidase; D006801:Humans; D006868:Hydrolysis; D007711:Klebsiella pneumoniae / Q000201:enzymology; D009822:Oils, Volatile / Q000032:analysis* / Q000737:chemistry; D010946:Plants, Medicinal; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet
D016588:Anticarcinogenic Agents; D047309:Flavones; D005419:Flavonoids; D009822:Oils, Volatile; D001192:Arylsulfatases; D005966:Glucuronidase; C023717:acacetin