摘要
建立了在线二维液相色谱法快速同时测定维生素AD制剂中维生素A和D的含量。依据药典对系统适应性的要求,选择一种极性嵌合的C_（18）柱（Accucore polar premium）作为二维色谱柱,再依据疏水减法模型原理和待测样品的脂溶性特点,选择C_8柱作为一维色谱柱,一维和二维分离均采用甲醇、乙腈和水作为流动相。样品经乙醇提取后直接进样分析,采用双梯度液相色谱右泵作为一维分析泵,完成含量较高的维生素A定量及维生素D的净化;采用左泵作为二维分析泵,维生素D及前维生素D在二维色谱柱上实现分离和定量。根据维生素D在一维色谱上的出峰起止时间,确定切割时间窗口,以500μL定量环收集含有目标物的馏分,检测波长分别为264 nm和325 nm。采用外标法完成对维生素D和前维生素D的定量。整个过程在密闭系统中自动化完成。维生素A在0.1~250 mg/L,维生素D在0.02~50.0 mg/L范围内线性关系较好,相关系数（r）大于0.999;维生素D的回收率为89.9%~98.9%,连续进样的精密度和重现性的RSD分别为0.48%和1.2%,表明方法的精密度和重现性较好。
A rapid method for the simultaneous determination of fat-soluble vitamins A and D in vita- min A and D pharmaceutical was developed using online two-dimensional liquid chromatography（2D - LC）. A type of polar embedded Cls was selected as the second dimension separating column accord- ing to system suitability testing requirement. Cs was chosen as first dimension separating column be- cause fat-soluble substance was rich in matrix and a better orthogonality could meet with polar embed- ded Cls according to hydrophobic subtraction model. Sample solution after extracted with methanol without further purification was injected into liquid chromatography directly. The purification of vita- min D and quantification of vitamins A acetate were accomplished simultaneously in one dimensional separation using the right pump of Dual Gradient LC （ DGLC ） with methanol - acetonitrile - water as mobile phase. Vitamin D and pre-vitamin D were separated and quantified in two dimensional column by using the left pump of DGLC. The eluted target from the first dimensional column （ 1-D column） was collected by a 500 IxL loop and then taken into the second dimensional column（2-D column） by the left pump of DGLC with high aqueous mobile phase. The cutting window could be confirmed by the retention time of target in one dimensional separation. Detection wavelengths of 325 nm and 264 nm were set at the diode-array detector. Vitamin A, vitamin D and pre vitamin D were quantified simultaneously and automatically in one injection by the external standard method. The results showed that the good linearities （ r 〉 0. 999 ） for vitamin A and vitamin D could be obtained in the range of 0. 1 - 250 mg/L and 0.02 - 50.0 mg/L, respectively. The recoveries of vitamin D ranged from 89.9% to 98.9%. The RSDs for precision and reproducibility are 0.48% and 1.2% , respec- tively. All the validating results indicated that vitamins A and D in pharmaceutical could be quanti- fied rapidly and accurately by this method.
出处
《分析测试学报》
CAS
CSCD
北大核心
2016年第1期28-34,共7页
Journal of Instrumental Analysis
关键词
二维液相色谱
在线分离
维生素D
维生素A
前维生素D
维生素AD滴剂
two-dimensional liquid chromatography（2D- LC）
online separation
vitamin D
vitamin A
pre vitamin D
vitamin A and D drops
