高效液相色谱-蒸发光散射检测器测定维生素D_3中间体

Determination of intermediates of vitamin D_3 by HPLC with evaporative light-scatter detector(ELSD)

为了同时测定胆固醇、胆固醇酯、7-酮胆固醇酯、7-脱氢胆固醇 4种维生素D3的中间体 ,研究确定了HPLC -蒸发光散射检测器测定的色谱分离条件。色谱条件为YWG -SiO2 柱 ;流动相 :正己烷 -四氢呋喃 (86∶14 ,V/V) ,流速 1mL/min ;进样量 2 0 μL ;漂移管温度 4 3℃ ,氮气流速 1.5L/min。在此条件下 ,4种组分可得到有效地分离 ,各物质色谱峰面积的自然对数与浓度的自然对数呈良好的线性关系 (r >0 .997) ,精密度RSD为 1.89%～ 2 .75 % ,最低检测限为 0 .1～ 0 .4 μg ,回收率测定值为 10 1.4 %～ 10 2 .8%。本法测定快速简便 ,结果准确。

An HPLC-ELSD method for the determination of intermediates of vitamin D_3 was studied.Chromatographic analysis was carried out on a YWG-SiO_2 column(4.6 mm×250 mm,5 μm) with hexane-tetrahydrofuran(86∶14, V/V)as the mobile phase.The relative standard deviation(RSD) was below 2.75% (n=6),and detection limits attainable were in the range 0.1 μg for cholesteryl acetate to 0.4 μg for 7-dehydrocholesterol, and the recoveries were from (101.4%) to 102.8%.The method is simple,rapid and accurate.