Determination of Thiamine and its Phosphate Esters by Electrophoresis and Fluorometry.

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Abstract
An improved modification for electrophoretic separation of thiamine and its phosphate esters is described. Electrophoresis is performed in 0.05 M sodium citrate buffer, pH 5.6, containing methanol, ethanol and propanol, instead of the 0.05 M acetate buffer, pH 3.8, used before, in which separation could not be readily obtained. With the citrate buffer for electrophoresis, the separation could be effected even if the samples contained perchloric acid and acetate, glycylglycine or phosphate buffer. The method is convenient for determining the activities of enzymes in the metabolism of thiamine compounds. The method was used for estimating the contents of thiamine and of thiamine mono-, di- and triphosphate esters in rat brain.