氟苯尼考对映异构体手性拆分及其光学纯度的测定

Separation of Florfenicol Enantiomer by Chiral Stationary Phase and Determination of Optical Purity

采用Chiralpak AD-H(4.6μm×250 mm,5μm)手性色谱柱,建立了氟苯尼考对映体的正相高效液相色谱拆分方法。考察了流动相中碱性添加剂、醇类改性剂种类及浓度对分离度、保留时间、理论塔板数、拖尾因子的影响。结果表明:以正己烷-异丙醇-甲醇(70∶15∶15)为流动相,流速为1 mL/min,柱温为30℃,检测波长为224 nm条件下,氟苯尼考与其光学异构体获得满意的分离效果。氟苯尼考在0.05～0.5g/L质量浓度范围内与其峰面积呈良好的线性关系,相关系数r为0.999 7;氟苯尼考的检出限为0.1μg/L;日内精密度RSD小于1.8%,日间精密度RSD小于2.3%;加标回收率为109%～112%,RSD不大于3.0%。该方法快速、方便,可用于工业生产中氟苯尼考光学纯度的控制。

A high performance liquid chromatographic （HPLC） method for the separation of florfeni- col enantiomers using a Chiralpak AD - H （4. 6 μm ×250 mm, 5 μm） was established. The influ- ences of the base additive, the kinds and concentrations of alcohol additives on the chiral separation of florfenicol including resolution, retention time, theoretical plate and tailing factor were investiga- ted. The results indicated that florfenicol was separated well from its optical isomer with n-hexane ： isopropyl alcohol ： methanol（70 ： 15 ： 15） as mobile phase at a flow rate of 1 mL . min-1 and a col- umn temperature of 30 ℃. The UV detection wavelength was set at 224 nm. The calibration curve for florfenicol was linear in the concentration range of 0. 05 - 0. 5 g . L- 1 with correlation coefficient of 0. 999 7. The limit of detection （LOD） was 0. 1 μg , L- 1. The intra-day relative standard deviations （RSD） were less than 1.8% and the inter-day RSDs were less than 2. 3%. The average spiked re- coveries were in the range of 109% - 112% with RSDs not more than 3.0%. The method was relia- ble and simple, and was suitable for the lality control of florfenicol in industrial production.