Determination of galactose and galactocerebroside using a galactose oxidase column and electrochemical detector.
A method has been developed to measure galactose and galactocerebroside using galactose oxidase immobilized on a solid resin. Galactose oxidase converts galactose and galactocerebroside to their corresponding aldehydes and hydrogen peroxide, the latter being electroactive and measurable by electrochemical detection using DC amperometric detection. The minimal detection limits of galactose and galactocerebroside were 1 and 2 microM, respectively. The linear response to galactose and galactocerebroside was to at least 300 microM. About 100 samples can be measured per hour using flow injection analysis. The activity of sulfatidase (cerebroside-3-sulfate-3-sulfohydrolase), which converts sulfatide (sulfogalactocerebroside) to galactocerebroside, was measured, and its inhibition by O-phospho-L-tyrosine was determined.
Stoecker
P W
PW
Department of Chemistry, Rutgers, State University of New Jersey, New Brunswick, New Jersey 08903, USA.
Manowitz
P
P
Harvey
R
R
Yacynych
A M
AM
D015374:Biosensing Techniques; D002553:Cerebroside-Sulfatase / Q000037:antagonists & inhibitors / Q000378:metabolism; D004563:Electrochemistry / Q000379:methods*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004800:Enzymes, Immobilized; D005690:Galactose / Q000032:analysis*; D005692:Galactose Oxidase; D005699:Galactosylceramides / Q000032:analysis*; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D019000:Phosphotyrosine / Q000494:pharmacology
D004791:Enzyme Inhibitors; D004800:Enzymes, Immobilized; D005699:Galactosylceramides; C002454:galactocerebroside; D019000:Phosphotyrosine; D005692:Galactose Oxidase; C093331:galactocerebroside sulfate sulfatidase; D002553:Cerebroside-Sulfatase; D005690:Galactose