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Clinical Chemistry 18: 503-508, 1972;
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Clinical Chemistry, Vol 18, 503-508, Copyright © 1972 by the American Association for Clinical Chemistry

Coulometric Determination of Activity of Acid or Alkaline Phosphatases in Serum

Marvin A. Brooks 1 and William C. Purdy 1

1 Department of Chemistry, University of Maryland, College Park, Md. 20742.

Department of Biochemistry and Drug Metabolism, Research Division, Hoffmann—La Roche Inc., Nutley, N. J. 07110.

A constant-current coulometric technique is described for measuring acid or alkaline phosphatase activity in serum. Electrogenerated bromine is used as the titrant of phenol enzymatically released from phenyl phosphate. A biamperometric end-point detection system is used, with an applied potential of 135 mV. Titrations are performed at pH 1 with use of a generating electrolyte of 0.5 molar KBr in 0.05 molar H2SO4. A procedure is described for determining serum alkaline phosphatase activity by use of 2-amino-2-methyl-1-propanol (0.25 mol/liter, pH 10.25) with Mg2+ activator (1 mmol/liter), and for determining serum acid phosphatase activity in citrate buffer (0.1 mol/liter, pH 4.9). Results compare favorably with those of the method of Kind and King [J. Clin. Pathol. 7, 322 (1954)].


Key Words: biamperometric end-point detection • comparison with results of Kind—King method • phenyl phosphate substrate • phenol titration

Submitted on December 14, 1971
Accepted on March 20, 1972






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Copyright © 1972 by the American Association for Clinical Chemistry.