Kimura M, Fujita T, Itokawa Y
Clin Chem. 1982 Jan;28(1):29-31.
A liquid-chromatographic method for determining the total thiamin content of blood is presented. Blood is deproteinized and incubated with Aspergillus oryzae carboxyl proteinase (EC 3.4.23.6; Takadiastase) to convert thiamin phosphate esters to free thiamin. An aliquot of the sample is applied to the column (Shodex OH-Pak M-414) of a high-performance liquid chromatograph. A 100 mg/L solution of potassium ferricyanide in 150 g/L sodium hydroxide is added to the column effluent with a proportioning pump, to convert thiamin into a fluorophore. The intensity of the fluorophore is measured with a spectrofluorophotometer and recorded graphically. The total thiamin in each blood sample appears as a single peak, and no co-eluting substance was detected. This method is simple, highly reproducible, and rapid, and its sensitivity is sufficient for determination of the thiamin content of 0.1-mL blood samples.
本文介绍了一种测定血液中硫胺素总量的液相色谱法。血液经去蛋白处理后，与米曲霉羧基蛋白酶（EC 3.4.23.6；胰淀粉酶）一起孵育，将硫胺素磷酸酯转化为游离硫胺素。取一份样品注入高效液相色谱仪的色谱柱（昭和电工OH-Pak M-414）。用比例泵将100 mg/L铁氰化钾溶液加入到150 g/L氢氧化钠溶液中，并将其添加到柱流出物中，将硫胺素转化为荧光团。用荧光分光光度计测量荧光团的强度并记录成图。每个血样中的总硫胺素呈现为一个单峰，未检测到共洗脱物质。该方法简单、重现性高、速度快，其灵敏度足以测定0.1 mL血样中的硫胺素含量。
