Liquid-chromatographic determination of vitamin K1 in plasma, with fluorometric detection.
Liquid-chromatographic determination of vitamin K1 in plasma, with fluorometric detection.
复制标题
液相色谱法测定血浆中维生素 K1，荧光检测。
作者：
Y. Haroon;D. Bacon;J. Sadowski
这种测定血浆中叶醌(维生素K1)的方法只需一次液相色谱步骤。与以往的检测方法相比，所需的血浆体积(0.5-1.0毫升)要小得多。用常规硅胶柱层析法提纯粗脂提取物，然后用正己烷/乙腈(体积比为1/4)的酸性混合物(每升含70 mmol氯化锌)溶解，提取粗脂肪提取物。通过添加金属锌将维生素K1转化为维生素K1对苯二酚后，将其选择性地提取到乙腈中。这个程序可以去除99%以上的污染油脂。在将维生素K1对苯二酚重新转化为维生素K1后，我们将脂类提取物直接进样到反相柱上。通过与内标(二氢维生素K1)的比较来定量维生素K1，并在柱后金属锌“在线”还原为对苯二酚后用荧光法进行测定。最终反相体系中维生素K1的检测下限约为0.05微克/L血浆，重复性的变异系数小于10%。22名健康空腹成人血浆维生素K1平均浓度为0.55(0.09~2.12)微克/L。
This assay for phylloquinone (vitamin K1) in plasma requires a single liquid-chromatographic step. Much smaller volumes of plasma (0.5-1.0 mL) are required than in previous assays. Before liquid chromatography, we purified crude lipid extracts by conventional chromatography on silica, then extracted the lipid fraction by dissolving it in an acidic mixture of hexane/acetonitrile (1/4 by vol) containing 70 mmol of zinc chloride per liter. The vitamin K1 was selectively extracted into acetonitrile after being converted to vitamin K1 hydroquinone by addition of zinc metal. This procedure removes greater than 99% of contaminating lipids. We injected the lipid extract directly onto a reversed-phase column after re-converting the vitamin K1 hydroquinone to vitamin K1. Vitamin K1 was quantified by comparison with the internal standard (dihydro-vitamin K1) and detected fluorometrically after post-column "on-line" reduction to the hydroquinone with zinc metal. The lower limit of detection for vitamin K1 in the final reversed-phase system was about 0.05 microgram/L plasma; CVs for replicates were less than 10%. The mean concentration of vitamin K1 in plasma from 22 healthy fasting adults was 0.55 (range 0.09-2.12) micrograms/L.
血清反式叶绿醌的高压液相色谱-还原电化学检测分析。
影响因子：
2.9
作者：
Ueno,T;Suttie,JW
通讯作者：
Suttie,JW