Fast method for the simultaneous determination of 2-oxo acids in biological fluids by high-performance liquid chromatography.
A fast and sensitive method for the single-run quantification of various 2-oxo acids including 2-ketoglutarate, glyoxylate and pyruvate is described. It ensures good separation of peaks with minor interference by other substances. The 2-oxo acid derivatives are measured photometrically at 324 nm after derivatization with phenylhydrazine and subsequent isocratic separation with ethanolic phosphate buffer on a C18 reversed-phase HPLC column. Recovery was found to be complete [range: 96.3 +/- 5.6% (pyruvate) to 104.8 +/- 5.2% (2-ketoglutarate)]. Detection limits ranged from less than 0.1 mumol/l (glyoxylate) to 0.25 mumol/l (2-ketoglutarate and pyruvate). Results for all substances examined showed good linearity with correlations (r2) of equal to or better than 0.998.
Lange
M
M
Department of Physiology, University of Kiel, Germany.
Mályusz
M
M
D002851:Chromatography, High Pressure Liquid; D006038:Glyoxylates / Q000032:analysis; D007202:Indicators and Reagents; D007651:Keto Acids / Q000032:analysis*; D007656:Ketoglutaric Acids / Q000032:analysis; D010659:Phenylhydrazines / Q000032:analysis; D011773:Pyruvates / Q000032:analysis; D012044:Regression Analysis; D013056:Spectrophotometry, Ultraviolet
D006038:Glyoxylates; D007202:Indicators and Reagents; D007651:Keto Acids; D007656:Ketoglutaric Acids; D010659:Phenylhydrazines; D011773:Pyruvates