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Continuous assay of serum 5' nucleotidase ec activity with imp as substrate and total automation using a transfer analyzer kem o mat

, : Continuous assay of serum 5' nucleotidase ec activity with imp as substrate and total automation using a transfer analyzer kem o mat. Journal of Clinical Chemistry and Clinical Biochemistry 21(8): 481-490

The continuous spectrophotometric assay of 5'-nucleotidase originally described by Heinz et al. (1980) was modified and fully automated on a Kem-O-Mat transfer analyzer, using IMP as substrate. The reaction product was H2O2 and the reduction of NADP was observed for 10 min at 340 nm and at a reaction temperature of C. The different factors involved in the enzyme reaction were checked, including the substrate concentration, reaction rate, linearity and substrate preservation. Normal values ranged from 1-13 U/l. Between-day reproducibility was estimated with 2 different commercial control sera, and the coefficient of variation was 5% for the upper limit of normal activity (23 U/l). There was good agreement between the present method and a semi-automatic colorimetric technique (for 100 [human] sera tested by both methods, r = 0.974 and the regression line equation, y = 0.85x - 1.5). Despite the lengthy reagent mixture preparation procedure, the method permitted assay of 50 samples/h. The occurrence of high serum blanks in certain pathological states is discussed.


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