Automated fluorimetric assay procedure for glucohydrolases using a routine centrifugal analyser assay of enzymes of lysosomal origin in plasma, II.
The manual fluorimetric procedure, considered as a reference method for the determination of N-acetyl-beta-D-glucosaminidase, beta-D-glucuronidase and beta-D-galactosidase in human plasma, was automated as a routine method, using the IL Monarch centrifugal analyser. Using a liquid 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 required only small volumes of undiluted plasma (4-10 microliters). This method appears to be reliable, sensitive, simple enough for routine analyses and as cost effective as the most common routine serum enzyme assays.
Goi
G
G
Department of Medical Chemistry and Biochemistry, Medical School, University of Milan, Italy.
Guagnellini
E
E
Bairati
C
C
Besozzi
M
M
Lombardo
A
A
Bollini
D
D
Lovagnini
A
A
Tettamanti
G
G
D000118:Acetylglucosaminidase / Q000097:blood*; D002498:Centrifugation / Q000379:methods; D005470:Fluorometry / Q000379:methods; D005966:Glucuronidase / Q000097:blood*; D006801:Humans; D008247:Lysosomes / Q000201:enzymology*; D012680:Sensitivity and Specificity; D001616:beta-Galactosidase / Q000097:blood*
D001616:beta-Galactosidase; D005966:Glucuronidase; D000118:Acetylglucosaminidase