目的 建立衍生化法分离（2R，4R）-4-甲基-2-哌啶甲酸乙酯酒石酸盐（MPFET）3种手性异构体。方法 以2，3，4，6-四-O-乙酰基-β-D-吡喃葡萄糖异硫氰酸酯（GITC）为柱前衍生化试剂，对MPFET手性异构体进行分离，并对衍生化条件进行优化。采用Venusil AQ C₁₈（250 mm×4.6 mm，5 μm）色谱柱作为固定相进行分离、监测和定量。以0.01 mol/L磷酸二氢钾溶液（用磷酸调pH至3.6）-乙腈（60∶40）为流动相，体积流量1.5 mL/min进行等度洗脱，采用紫外检测器，检测波长266 nm；柱温30℃；进样量20 μL。结果 MPFET与2S，4S-异构体分离度为1.76。2S，4R-异构体的线性范围为1.500～8.999 μg/mL，2R，4S-异构体的线性范围为0.255 2～1.531 0 μg/mL，2S，4S-异构体的线性范围为0.250 1～75.000 0 μg/mL，MPFET的线性范围为0.250 1～600.100 0 μg/mL，回收率均在90%～108%内，RSD均不大于3.0%。结论 柱前衍生化法分离MPFET中3种手性异构体，专属性强、准确度高、灵敏度高、重复性好，可用于MPFET手性异构体的分离和质量控制。

Objective To establish a derivatization method for the separation of three chiral isomers of (2R, 4R)-4-methyl-2-piperidine formate ethyl tartrate (MPFET). Methods 2, 3, 4, 6-tetra-o-acetyl was used-β-D-glucopyranose isothiocyanate (GITC) was used as pre column derivatization reagent to separate the chiral isomers of MPFET, and the derivatization conditions were optimized. Use venusil AQ C₁₈ (250 mm×4.6 mm, 5 μ m) chromatographic column is used as stationary phase for separation, monitoring and quantification. 0.01 mol/L potassium dihydrogen phosphate solution (adjust pH to 3.6 with phosphoric acid)-acetonitrile (60:40) was used as mobile phase, and the volume flow was 1.5 mL/min for isocratic elution. UV detector was used with detection wavelength of 266 nm, column temperature 30℃, injection volume 20 μL. Results The separation of MPFET from 2S, 4S isomers was 1.76. The linear range of 2S, 4R isomers was 1.500-8.999 μg/mL, the linear range of 2R, 4S isomers was 0.255 2-1.531 0 μg/mL, the linear range of 2S, 4S isomers was 0.250 1-75.000 0 μg/mL, the linear range of MPFET was 0.250 1-600.100 0 μg/mL, the recoveries were within 90%-108%, and the RSD was not greater than 3.0%. Conclusion Precolumn derivatization has strong specificity, high accuracy, high sensitivity and good repeatability. It can be used for the separation and quality control of chiral isomers in MPFET.

R914

中国医学科学院医学与健康科技创新工程项目资助（2019-I2M-5-020）；天津市科技计划项目（18YFZCSY00090）