天然类视黄醇的梯度反相高效液相色谱分离

Gradient reversed-phase high-performance liquid chromatographic separation of naturally occurring retinoids.

作者信息

Furr H C, Amédée-Manesme O, Olson J A

出版信息

J Chromatogr. 1984 Aug 10;309(2):299-307. doi: 10.1016/0378-4347(84)80037-7.

Abstract

A gradient reversed-phase high-performance liquid chromatographic technique is described for the facile separation and quantitation of the naturally occurring retinoids: retinoic acid, retinol, and retinyl esters. An octadecylsilane column (Waters mu Bondapak C18) is used, with gradient elution from methanol--water (80:20) (solvent A) to 70% or 100% methanol--tetrahydrofuran (50:50) (solvent B) at 2.0 ml/min; detection is by absorbance at 325 nm. Analysis can be completed, with return to starting conditions, in 25-30 min. The method is inherently flexible: retinyl esters can be eluted as a group, with little resolution, by gradient to 100% solvent B, or mostly resolved by gradient to 70% solvent B; separation of retinoids more polar than retinoic acid can be achieved by use of greater proportions of water in solvent A. The separation of vitamin A compounds from extracts of human, rat, and pig liver and from rat kidney by this technique is described.

摘要

描述了一种梯度反相高效液相色谱技术，用于轻松分离和定量天然存在的类维生素A：视黄酸、视黄醇和视黄酯。使用十八烷基硅烷柱（沃特世μ Bondapak C18），以2.0 ml/min的流速从甲醇 - 水（80:20）（溶剂A）梯度洗脱至70%或100%甲醇 - 四氢呋喃（50:50）（溶剂B）；通过在325 nm处的吸光度进行检测。在25 - 30分钟内即可完成分析并回到起始条件。该方法本质上具有灵活性：视黄酯可通过梯度至100%溶剂B作为一组洗脱，分辨率较低，或通过梯度至70%溶剂B大部分得到分离；通过在溶剂A中使用更大比例的水，可以实现比视黄酸极性更强的类维生素A的分离。描述了用该技术从人、大鼠和猪肝提取物以及大鼠肾脏中分离维生素A化合物的方法。