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Abstract
A simple and rapid isocratic high-performance liquid chromatographic (HPLC) method has been developed for the determination of seven alpha-keto acids, glyoxylic acid, pyruvic acid, 2-oxobutyric acid, 3-methyl-2-oxobutyric acid, 2-oxoglutaric acid, 2-oxohexanoic acid, and phenylpyruvic acid using meso silbenediamine as derivatizing reagent in the blood samples. The chromatographic separation was achieved with methanol-water-acetonitrile-tetrahydrofuran (38.4:60:1:0.6, v/v/v/v) as the mobile phase, a C18 column, and UV detection at 255 nm. Linear calibration curves were obtained with 0.2 to 100 µg/mL. The limits of detection and quantitation were obtained within the range 0.07–0.2 µg/mL and 0.21–0.6 µg/mL, respectively. The method was applied for the determination of alpha-keto acids from human serum and urine of uremic and diabetic patients before hemodialysis, after hemodialysis, and healthy volunteers.
ACKNOWLEDGMENT
We thank G. M. Mastoi (University of Sindh, Jamshoro) for his support in buying materials for this research work.
Notes
*Data presented as mean (n = 3), and relative S.D. (%) are shown within the brackets.
*M = Male, F = Female, **%RSD, n = 6.
*RSD, n = 6.
*RSD, n = 6.
*RSD, n = 6.