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Clinical Chemistry 28: 2206-2213, 1982;
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Clinical Chemistry, Vol 28, 2206-2213, Copyright © 1982 by American Association for Clinical Chemistry

A continuous-flow (AutoAnalyzer II) procedure for measuring erythrocyte transketolase activity

PP Waring, D Fisher, J McDonnell, EL McGown and HE Sauberlich

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.


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