Clinica Chimica Acta 2002-12-01

Continuous monitoring of prostatic acid phosphatase using self-indicating substrates.

Klaus Lorentz

Index: Clin. Chim. Acta 326(1-2) , 69-80, (2002)

Full Text: HTML

Abstract

The continuous measurement of acid phosphatase (EC 3.1.3.2) activity in serum represents an analytical task not yet sufficiently accomplished.Introducing two novel substrates-2-chloro-4-nitrophenyl phosphate (CNP-P), which was preferred, and 4-nitronaphthyl-1-phosphate (NN-P)-an alternative assay to measure enzymatic activity was developed and compared with a modification of Hillmann's method (azo coupling of released naphth-1-ol with a diazonium compound). Apart from different substrate concentrations of 2-chloro-4-nitrophenyl phosphate, 4 mmol/l, and naphthyl-1-phosphate (N-P), 8 mmol/l (with Fast Red TR, 5 mmol/l), respectively, following identical conditions were selected: Citrate, 50 mmol/l, pH 5.75; pentane-1,5-diol, 150 mmol/l; tartrate, 60 mmol/l; 37 degrees C.Whereas intensity and stability of the azo dye unpredictably depend on the albumin concentration of the sample, the direct test with 2-chloro-4-nitrophenyl phosphate resisted sample interferences, showed no intrinsic hydrolysis by albumin, relied on stable reagents and proved superior in sensitivity, precision and ease of handling. In measuring prostatic phosphatase, the proposed procedure closely correlated with Hillmann's method. The preliminary 0.95-reference intervals for adults were 1.2-3.9 kU/l and 5.8-14.8 U/l for total activity, respectively.The direct assay of the enzyme is suited as an economic, rapid and robust method for mechanized or manual use.