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Clinical Chemistry 21: 1619-1626, 1975;
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Clinical Chemistry, Vol 21, 1619-1626, Copyright © 1975 by the American Association for Clinical Chemistry

Reaction-Rate Assay of Serum Sorbitol Dehydrogenase Activity at 37 °C

Clive I. Rose 1 and Arthur R. Henderson 1

1 Department of Clinical Biochemistry, University Hospital (University of Western Ontario), 339 Windermere Rd., London, Ontario N6G 2K3, Canada

The sorbitol dehydrogenase (EC 1.1.1.14) activity of human serum and liver was investigated at 37 °C. Various buffers were examined but tris(hydroxymethyl)aminomethane HCl in a final concentration of 90 mmol/liter (pH 6.6, at 37 °C) was the one with which activity was the greatest. The enzyme is inhibited by its substrate, D-fructose, and activity was greatest with a substrate concentration (in the final assay mixture) of 500 mmol/ liter and an NADH concentration of 247 µmol/liter. The specimen volume was 100 µl. The enzyme from liver and serum was shown to have similar Km values and activation energies, and we conclude that the liver enzyme was unchanged on passing into the extrahepatic space. Normal values for this assay are 0-2.6 U/liter (2.5 and 97.5 percentiles)


Key Words: liver disease • Km value • optimum conditions for assay • reference values • hepatic anoxia

Submitted on June 10, 1975
Accepted on July 22, 1975




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