Erythrocytic glucose-6-phosphate dehydrogenase measured by a differential pH technique.
We propose a new quantitative electrochemical method for determining glucose-6-phosphate dehydrogenase (EC 1.1.1.49) activity in purified erythrocytes or in whole blood, based on measurement of the pH change caused by oxidation of glucose 6-phosphate to 6-phosphogluconic acid, with simultaneous reduction of NADP+ to NADPH + H+. No sample pretreatment (e.g., preparation of hemolysate) is needed, and the automatic correction for sample blanks obviates interference from 6-phosphogluconate dehydrogenase (EC 1.1.1.44). The method is simple and fast, and the standard curve is linear to at least 2200 U/L at 37 degrees C. The within-day CV was 3.9% for activities in healthy individuals (mean value 1204 U/L), and 10% for deficient ones (classified as belonging to class II, mean value 407 U/L). Results (y) correlated well with those obtained with the WHO-recommended method (x): y = 1.13x + 0.02 (r = 0.971) for purified erythrocytes. Normal reference intervals are reported for the enzyme in purified erythrocytes and in whole blood.
D004912:Erythrocytes / Q000201:enzymology*; D005954:Glucosephosphate Dehydrogenase / Q000097:blood*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008433:Mathematics; D008722:Methods; D009249:NADP; D010734:Phosphogluconate Dehydrogenase / Q000097:blood; D012016:Reference Values
D009249:NADP; D010734:Phosphogluconate Dehydrogenase; C029757:phosphogluconate dehydrogenase (decarboxylating); D005954:Glucosephosphate Dehydrogenase