正相-反相两步高效液相色谱法制备毫克量级维生素D_3异构体

利用正相半制备柱和反相分析柱的两步高效液相色谱法,在实验室规模制备了毫克是级维生素D_3的三个重要异构体——维生素D_3前体、光甾醇_3和速甾醇_3·维生素D_3原体,7-去氢胆固醇的光照是在一个装有UVB荧光灯的光治疗室中进行。采用紫外分光光度法和高效液相色谱法对制得的三个异构体进行了定性定量分析。在这三个异构体检测极限下,除了维生素D_3前体中有不超过0.25％的维生素D_3杂质外,三个产品中均没有发现任何除它本身之外的其他异构体杂质。

PREPARATION OF MILLIGRAM QUANTITY OF VITAMIN D_3 ISOMERS BY A TWO-STEP HIGH PERFORMANCE LIQUID CHROMATOGRAPHIC METHOD

The preparation of milligram quantities of three vitamin D_3 isomers, pre-vitamin D_3, lumisterol_3 and tachysterol_3, were carried out in a laboratory scale first by irra-diating 7- dehydrocholesterol in a phototherapy chamber equipped with UVB lamps, fol-lowed by using a two- step high performance liquid chromatography (HPLC) with asemi - preparative normal phase column and an analytical reverse- phase column. The finalproducts obtained were identified by UV spectrophotometry and HPLC. According to thedetection limits for the three isomers, no contamination with any other isomers was de-tected in the previtamin D_3, lumisterol_3 and tachysterol_3 preparations, except that a verysmall amount of vitamin D_3, constituting no more than 0.25% of the product, was foundin previtamin D_3 preparation.