Microdetermination of galactose and galactose 1-phosphate in dried blood spots |
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| Authors: | Yushin Fujimura Sumikazu Ishii Masahiko Kawamura Hiroshi Naruse |
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| Institution: | 1. Nagoya City Public Health Institute, Nagoya, Japan;2. Division of Early Detection of Handicapped, National Center for Nervous, Mental and Muscular Disorders, Tokyo, Japan;3. Meijyo Hospital, Nagoya, Japan |
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| Abstract: | Newborn screening for galactosemia (galactose-1-phosphate uridyltransferase deficiency), as well as for other defects in galactose metabolism (galactokinase deficiency and uridine diphosphogalactose 4-epimerase deficiency), requires a method of determining both galactose and galactose 1-phosphate in dried blood. We have developed a sequential quantitative method for the microdetermination of galactose and galactose 1-phosphate that can be applied to 3-mm-diameter disks of dried blood and that can be used with a Technion Autoanalyser II equipped with a fluorometer.Galactose is determined by the fluorescence of NADH following treatment with β-galactose dehydrogenase and with the consequent reduction of NAD. The complete system includes alkaline phosphatase for the hydrolysis of galactose 1-phosphate, so that the total amounts of a galactose and galactose 1-phosphate are determined. For the measurement of galactose alone, alkaline phosphate is omitted from the system. The difference in fluorescence between that from the complete system and that from the alkaline phosphatase-omitted system yields the concentration of galactose 1-phosphate. |
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| Keywords: | To whom correspondence should be addressed |
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