AUTOMATED FLUOROMETRIC ASSAY PROCEDURE FOR GLUCOHYDROLASES USING A ROUTINE CENTRIFUGAL ANALYZER ASSAY OF ENZYMES OF LYSOSOMAL ORIGIN IN PLASMA .2.

The manual fluorimetric procedure, considered as a reference method fo r the determination of N-acetyl-beta-D-glucosaminidase, beta-D-glucuro nidase and beta-D-galactosidase in human plasma, was automated as a ro utine method, using the IL Monarch centrifugal analyser. Using a liqui d standard with a known enzyme content, the automated assay correlated fairly well with the reference manual method (r values very close to 1). Its analytical imprecision was much lower than that of the manual method. The automated assay of N-acetyl-beta-D-glucosaminidase, beta-D -glucuronidase and beta-D-galactosidase gave coefficients of variation of 5.7-6.9, 3.6-5.0 and 3.8-4.2%, respectively, detection limits of 4 , 2 and 1 mU/l plasma respectively, and linear responses of up to 73, 8.4 and 0.9 U/l of plasma respectively. Furthermore, the method requir ed only small volumes of undiluted plasma (4-10 mu l). This method app ears to be reliable, sensitive, simple enough for routine analyses and as cost effective as the most common routine serum enzyme assays.