高速液体クロマトグラフィーによる血液中総ビタミンB_1定量法

概要

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A high sensitive method for measuring total thiamine by high performance liquid chromatography (HPLC) was presented. Blood (100μl) was deproteinized and the supernatant was adjusted to pH 4.5 with CH_3COONa. Then diastase was added and incubated 37℃ for 8〜10 hours. For HPLC, the mobile phase (0.2M NaH_2PO_4) was eluted at a flow-rate of 0.5 ml/min. A 50〜100μl of sample was loaded on the sample loop and then injected on to the column. Potassium ferricyanide (0.01%) in sodium hydroxide (15%) solution was applied at 0.5ml/min by a proportioning pump and mixed with the column effluent to convert thiamine into thiochrome. Thiochrome was measured with a spectrofluorometer connected to the chromatograph and recorded graphically. Using this method, thiamine concentrations in blood of humans and animals were determined. Average thiamine concentrations in blood were 4.25μg/dl (n=10), 24.33 μg/dl (n=7) and 25.78μg/dl (n=5) in humans, rats and rabbits, respectively. Reliability and sensitivity of this method are satisfactory.
日本ビタミン学会の論文
1981-04-25