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Summary: A simple method is described for the simultaneous determination of alkaline phosphatase (EC 3.1.3.1) and 5'-nucleotidase (EC 3.1.3.5) iri serum. The method is based on the determination of inorganic phosphorus released by the action of the two enzymes on adenosine-S'-monophosphate at pH 9.5 (200 mmol/1 tris-buffer) in the presence and absence of ,-cysteine. This amino acid at a concentration of 2-10 mmol/1 was found to be a specific inhibitor for alkaline phosphatase but with no effect on 5'-nucleotidase activity.

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