High-performance liquid chromatographic determination of alpha-keto acids in human serum and urine using 1,2-diamino-4,5-methylenedioxybenzene as a precolumn fluorescence derivatization reagent.
A simple and highly sensitive high-performance liquid chromatographic (HPLC) method for the determination of alpha-keto acids in human serum and urine is described. In an acidic solution, twelve species of alpha-keto acids examined were converted by reaction with 1,2-diamino-4,5-methylenedioxy-benzene into highly fluorescent derivatives. The derivatives were separated isocratically by reversed-phase HPLC on a TSK gel ODS-80TM column and detected fluorimetrically. Eight alpha-keto acids in human serum and eleven alpha-keto acids in human urine can be determined simultaneously. The detection limits (signal-to-noise ratio = 5) are 6-44 fmol in an injection volume of 5 microliters. The intra-assay relative standard deviations for both serum and urine sample analyses are usually ca. 5%.
Wang
Z J
ZJ
Faculty of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
Zaitsu
K
K
Ohkura
Y
Y
D002851:Chromatography, High Pressure Liquid; D005260:Female; D005456:Fluorescent Dyes; D006801:Humans; D007651:Keto Acids / Q000032:analysis* / Q000097:blood / Q000652:urine; D008297:Male; D010655:Phenylenediamines; D012996:Solutions; D013050:Spectrometry, Fluorescence
D005456:Fluorescent Dyes; D007651:Keto Acids; D010655:Phenylenediamines; D012996:Solutions; C058886:1,2-diamino-4,5-methylenedioxybenzene