A continuous-flow (AutoAnalyzer II) procedure for measuring erythrocyte transketolase activity.
In this semiautomated method, an AutoAnalyzer II is used to measure the enzymic production of glyceraldehyde 3-phosphate in hemolysates, to assay erythrocyte transketolase (EC 2.2.1.1) activity. Hemolysate and indicator reactions are separated by dialysis to eliminate hemoglobin interference and increase sensitivity. Internal standards of glyceraldehyde 3-phosphate in hemolysate carriers were quantitatively measured with good precision and accuracy in the presence or absence of the transketolase substrate, ribose 5-phosphate. Chart-recorder values for these standards were used to calibrate the AutoAnalyzer output in IUB units (U) of transketolase activity. Substrate-product relationships were examined to characterize reaction kinetics and optimize assay conditions. AutoAnalyzer transketolase results correlated well with those from two manual procedures.
D001322:Autoanalysis / Q000379:methods; D002138:Calibration; D004912:Erythrocytes / Q000201:enzymology*; D005986:Glyceraldehyde 3-Phosphate / Q000096:biosynthesis; D006801:Humans; D012016:Reference Values; D012267:Ribosemonophosphates; D014174:Transketolase / Q000097:blood*
D012267:Ribosemonophosphates; D005986:Glyceraldehyde 3-Phosphate; D014174:Transketolase