Automated determination of acid phosphatase. II. Technic for using robot chemist.

Abstract

A procedure is presented for the automated determination of acid phosphatase activity in biological materials using the Robot Chemist. Although either phenylphosphate and a-naphthylphosphate may be used as substrate in this analysis, the procedure is described in detail for serum acid phosphatase using a-naphthylphosphate in 0.1 M citrate, pH 5.2, since this substrate is more selective for prostatic acid phosphatase in human serum. Enzymically generated a-naphthol is determined by the Emerson reaction (alkaline aminoantipyrine and ferricyanide), modified for use with this auto-mated system. Correlations are presented between the results obtained on the Robot Chemist and the identical procedure developed for the AutoAnalyzer. T1HIS PAPER reports the development of a simplified procedure using the Robot Chemist * for the automated determination of acid phospha-tase activity which is applicable to the assay of biologic material of either plant or animal origin. Either phenylphosphate or -naphthyl-phosphate may be used as substrate for this assay. A detailed descrip-tion of the analysis is given using -naphthylphosphate in 0.1 M citrate, pH 5.2, since this substrate is more selective for prostatic acid phos-phatase in human serum (1, 2). The procedure is based on measure-ment of the absorbance of the color produced by the Emerson reaCtiOll with alkaline aminoantipyrine and ferricyanide on the enzymically gen-erated -naphthol (3). The procedure is an adaptation of a similar procedure, recently developed, using the AutoAnalyzert (4).