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.
Jagiellonian University, Institute of Molecular Biology, Kraków, Poland.
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Kozik
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Wróbel
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Turyna
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