UPLC-MS/MS方法测定盐酸哌替啶原料及注射液中的神经毒性杂质1-甲基-4-苯基-1,2,3,6-四氢吡啶

目的:建立UPLC-MS/MS方法,定量分析盐酸哌替啶原料及注射液中的神经毒性杂质——1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)。方法:采用ACQUITY CSH Phenyl-Hexyl(2.1 mm×100 mm, 1.7 μm)色谱柱进行色谱分离,流动相位0.1%甲酸水-乙腈,梯度洗脱;以三重四极杆作为检测器,采用多反应监测(MRM)扫描方式,选择母离子(m/z 174)→子离子(m/z 44)作为MRM定量离子对。用该方法检测盐酸哌替啶原料药及其注射液中MPTP,MPTP的含量按峰面积以外标法计算。结果:MPTP质量浓度在0.25~201.4 ng·mL^-1范围内线性关系良好(r=0.999 6);平均加样回收率(n=9)为102.5%,RSD为1.5%;供试品溶液在24 h内的稳定性良好(RSD=1.6%);MPTP的定量限1.93×10^-2 ng·mL^-1,检出限为0.58×10^-2 ng·mL^-1;3批原料药中MPTP含量为175~185 ng·g^-1,10批注射液中MPTP含量为6.3~33.2 ng·mL^-1。结论:本方法适用于盐酸哌替啶原料药及注射液中MPTP的含量测定。

Objective:To establish a UPLC-MS/MS method for reliable quantitative analysis of 1-methyl-4-pheyl-1,2,3,6-tetrahydropyridine(MPTP) in pethidine hydrochloride active pharmaceutical ingredient(API) and injection.Methods:An ACQUITY CSH Phenyl-Hexyl(2.1 mm×100 mm,1.7 μm) column was adopted to achieve chromatographic separation with gradient elution.T mobile phase consisted of water containing 0.1% formic acid and acetonitrile.The triple quadrupole is operated in the multi reaction monitoring(MRM) mode,selecting the m/z 174→m/z 44 as the quantitative ion pair.Pethidine hydrochloride API and injection from pharmaceutical companies was determined by the method,and the content computation was performed with external standard carve method.Results:The linear range of MPTP was 0.25-201.4 ng·mL^-1 with the correlation coefficient (r=0.999 6).The average recovery(n=9) was 102.5%,RSD=1.5%.The sample solution was stable within 24 h(RSD=1.6%).The limit of quantitation of MPTP was 1.93×10^-2 ng·mL^-1 and the limit of detection was 0.58×10^-2 ng·mL^-1.The contents of MPTP in 3 API samples were 175-185 ng·g^-1,and the contents of MPTP in 10 injection samples were 6.3-33.2 ng·mL^-1.Conclusion:The established method can be applied to the content determination of MPTP in pethidine hydrochloride API and injection.

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