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Clinical Chemistry 20: 1349-1352, 1974;
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Clinical Chemistry, Vol 20, 1349-1352, Copyright © 1974 by the American Association for Clinical Chemistry

Measurement of Glucose-6-phosphate Dehydrogenase and 6-Phosphogluconate Dehydrogenase Activites in Erythrocytes by Use of a Centrifugal Analyzer

John F. Nicholson 1, Selma H. Bodourian 1, and Michael A. Pesce 1

1 Department of Pediatrics, College of Physicians and Surgeons of Columbia University, and the Microchemistry Laboratory, Babies Hospital, the Children's Medical and Surgical Center of New York, New York, N. Y. 10032.

We describe an accurate automated method for measuring activity of glucose-6-phosphate dehydrogenase (EC 1.1.1.49) in erythrocytes with a centrifugal analyzer. Blood is collected in microhematocrit tubes, centrifuged, and the erythrocytes are lysed with digitonin. Glucose-6-phosphate dehydrogenase activity is determined by mixing the hemolysate with glucose-6-phosphate, NADP+, and 6-phosphogluconate dehydrogenase #{ 233} (EC 1.1.1.44) in triethanolamine—EDTA buffer at pH 7.6, and measuring the rate of NADPH production for 3 min. Under these conditions 2 mol of NADPH are produced per mole of glucose-6-phosphate oxidized, ensuring the accuracy of the method and increasing its sensitivity. Activity is referred to hemoglobin, measured as cyanmethemoglobin. Activity of glucose-6-phosphate dehydrogenase added to hemolysates was well accounted for. Results obtained by our method and by the method of Bishop [J. Lab. Clin. Med. 68, 149 (1966)] are virtually identical. Our method requires a small amount of blood and is accurate and rapid; thus it is well suited for use in surveying large populations for glucose-6-phosphate dehydrogenase deficiency. A simple modification of the method may be used to determine the activity of 6-phosphogluconate dehydrogenase.


Key Words: screening • CentrifiChem • inherited disorders • normal values

Submitted on June 26, 1974
Accepted on August 5, 1974






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Copyright © 1974 by the American Association for Clinical Chemistry.