Competitive protein-binding radioassay of thiamine in simple solutions and in multivitamin pharmaceuticals.
A competitive inhibition radioassay of thiamine is described using a gel obtained by coupling a buckwheat-seed thiamine-binding protein to CNBr-activated Sepharose. The sample to be analysed is incubated with gel suspension and [14C]thiamine and after centrifugation the radioactivity of the supernatant is measured. The method is simple and specific, and applicable over a thiamine concentration range 0.5-5 microM with a coefficient of variation typically below 5%. The gel is reusable and stable for several months. Applicability of the method for direct determination of thiamine in multivitamin pharmaceuticals is demonstrated.
Kozik
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Jagiellonian University, Institute of Molecular Biology, Kraków, Poland.
Wróbel
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Turyna
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D001667:Binding, Competitive; D002352:Carrier Proteins; D011485:Protein Binding; D012685:Sepharose; D013050:Spectrometry, Fluorescence; D013831:Thiamine / Q000032:analysis*; D014815:Vitamins / Q000032:analysis*
D002352:Carrier Proteins; D014815:Vitamins; C026989:thiamine-binding protein; D012685:Sepharose; D013831:Thiamine