目的 建立衍生化法分離（2R，4R）-4-甲基-2-哌啶甲酸乙酯酒石酸鹽（MPFET）3種手性異構(gòu)體。方法 以2，3，4，6-四-O-乙?；?β-D-吡喃葡萄糖異硫氰酸酯（GITC）為柱前衍生化試劑，對(duì)MPFET手性異構(gòu)體進(jìn)行分離，并對(duì)衍生化條件進(jìn)行優(yōu)化。采用Venusil AQ C₁₈（250 mm×4.6 mm，5 μm）色譜柱作為固定相進(jìn)行分離、監(jiān)測(cè)和定量。以0.01 mol/L磷酸二氫鉀溶液（用磷酸調(diào)pH至3.6）-乙腈（60∶40）為流動(dòng)相，體積流量1.5 mL/min進(jìn)行等度洗脫，采用紫外檢測(cè)器，檢測(cè)波長(zhǎng)266 nm；柱溫30℃；進(jìn)樣量20 μL。結(jié)果 MPFET與2S，4S-異構(gòu)體分離度為1.76。2S，4R-異構(gòu)體的線性范圍為1.500～8.999 μg/mL，2R，4S-異構(gòu)體的線性范圍為0.255 2～1.531 0 μg/mL，2S，4S-異構(gòu)體的線性范圍為0.250 1～75.000 0 μg/mL，MPFET的線性范圍為0.250 1～600.100 0 μg/mL，回收率均在90%～108%內(nèi)，RSD均不大于3.0%。結(jié)論 柱前衍生化法分離MPFET中3種手性異構(gòu)體，專(zhuān)屬性強(qiáng)、準(zhǔn)確度高、靈敏度高、重復(fù)性好，可用于MPFET手性異構(gòu)體的分離和質(zhì)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

中國(guó)醫(yī)學(xué)科學(xué)院醫(yī)學(xué)與健康科技創(chuàng)新工程項(xiàng)目資助（2019-I2M-5-020）；天津市科技計(jì)劃項(xiàng)目（18YFZCSY00090）