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Colorimetric determination of galactose and galactose-1-phosphate from dried blood

โœ Scribed by Frank Diepenbrock; Rolf Heckler; Haribert Schickling; Thomas Engelhard; Daniel Bock; Johannes Sander

Publisher
Elsevier Science
Year
1992
Tongue
English
Weight
240 KB
Volume
25
Category
Article
ISSN
0009-9120

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โœฆ Synopsis

A colorimetric microassay for the simultaneous quantitative determination of galactose (Gal) and galactose-l-phosphate (Gall-P) in dried blood spots is described. An enzymatic reaction involving alkaline phosphatase (EC 3.1.3.1) and galactose dehydrogenase (EC 1.1.1.48) produces NADH, which is coupled with diaphorase (EC 1.8.1.4) and iodonitrotetrazolium violet (INT). The colourless INT is converted to a formazan of red colour the intensity of which is quantitated either photometrically by a microplate reader or determined visually with sufficient sensitivity for screening purposes. We evaluated the assay on 200,000 blood samples in a newborn screening program, and were able to distinguish between classical and milder forms of galactosemia with ease.

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