柱前衍生高效液相色谱法测定四氢噻唑-2-硫酮-4-羧酸的光学纯度

用苯胺对四氢噻唑-2-硫酮-4-羧酸(TTCA)进行柱前衍生,将其衍生物在手性固定相上拆分,通过二极管阵列紫外检测器和在线旋光检测仪对其衍生物进行检测,建立了一种拆分TTCA消旋体、测定TTCA光学纯度的新方法。以正己烷和乙醇或异丙醇为流动相,在DNB-Leucine手性固定相上对TTCA衍生物进行了拆分,并考察了流动相组成和柱温对其衍生物分离的影响,获得较优分析条件,分离因子大于1.2。非手性试剂苯胺柱前衍生化法测定(R)-TTCA的光学纯度与旋光度方法比较,结果一致,相对偏差小于1.34%。

Determination of Enantiomeric Purity of 2-Thiothiazolidine-4-carboxylic Acid by High Performance Liquid Chromatography with Pre-Column Derivatization

The 2-thiothiazolidine-4-carboxylic acid was derivatized with aniline that was the reagent of pre-column derivatization of high performance liquid chromatography. The derivative was separated by DNB-Leucine chiral stationary phase, and detected by Photo-Diode-Array detector and Optical-Rotation detector. A method for separating the racemate and determining the enantiomeric purity of 2-thiothiazolidine-4-carboxylic acid was established. Chromatographic model was employed for enantiomeric separation of 2-thiothiazolidine-4-carboxylic acid derivatives on the DNB-Leucine chiral column in hexane/alcohol (ethanol or isopropyl alcohol) mobile phase. The effects of chromatography conditions, such as mobile phases and temperature to chiral recognition, had been investigated. A better condition of analysis was obtained and the separation factor was more than 1.2. Furthermore, the results of enantiomeric purity determined by both polarimeter and HPLC were very consistent and the relative deviation was less than 1.34%. The method of pre-column derivatization of high performance liquid chromatography is credible, accurate and can be applied to determine enantiomeric purity of 2-thiothiazolidine-4-carboxylic acid.