建立一种超高效合相色谱法（UPC2）快速分离和测定氟苯尼考对映体的方法。采用Acquity Trefoil CEL2（150 mm×3.0 mm，2.5 µm）手性色谱柱，CO2和甲醇为流动相，梯度洗脱，流速1.0 mL/min，检测波长为224 nm，进样体积5 µL，系统背压17.2 MPa，柱温40 ℃。实验结果表明：两种氟苯尼考对映体在4.00~400.00 mg/L线性范围内与峰面积呈良好线性关系，相关系数均（r2）在0.9993以上，仪器检出限（LOD，S/N=3）为2.0 mg/L。连续重复进样10.0 mg/L混合标准工作溶液6次，(-)、(+)-氟苯尼考的峰面积相对标准偏差值（RSD，n=6）分别为0.65%、0.81%。应用该方法对市售氟苯尼考标准品进行了分离及测定，结果显示，3份标准品均未检出(+)-氟苯尼考，均检出(-)-氟苯尼考，含量为9.20~10.00 mg/L，在6.0 min内完成了两个氟苯尼考对映体的分离及含量测定，该方法检测速度快、分离度高、绿色环保，可以满足氟苯尼考对映体的分离检测要求。

A new method was established for the separation and determination of two florfenicol enantiomers using ultra-performance convergence chromatography (UPC2). The chromatographic separation conditions were optimized, and the stability of standard solutions of (-)-florfenicol and (+)-florfenicol enantiomers were observed. The separation process was carried out in Waters Acquity Trefoil CEL2 (150×3.0 mm, 2.5 µm) chiral columns. The mobile phase used carbon dioxide and methanol at a flow rate of 1.0 mL/min with gradient elution. The UV detection wavelength was set to 224 nm, the injection volume was 5 µL, the system backpressure was 17.2 MPa, and the column temperature was maintained at 40 ℃. The method validation results indicate that the limit of detection (LOD, S/N=3) for both enantiomers was 2.00 mg/L, and the calibration linear range was between 4.00 and 400.00 mg/L, with correlation coefficients (r2) > 0.9993. Repeated measurements of the peak area using 10 mg/L standard solutions of each florfenicol enantiomer showed relative standard deviations (RSD, n=6) of 0.65 and 0.81%, respectively. This method was then used to separate and measure commercially available standard florfenicol racemates. The results indicate that (+)-florfenicol was not detected in the three standard florfenicol racemates, but (-)-florfenicol (9.20~10.00 mg/L) was detected in all of them. The separation and content determination of the two florfenicol enantiomers were completed within 6.0 min. Therefore, this is a rapid, high-resolution, eco-friendly method that can be used for the separation and determination of florfenicol enantiomers.

国家重点研发计划（2019YFC1605400)；海关总署科研项目(2020HK196)；浙江省科技计划项目(2018C37014)