UHPLC-MS法测定2种硫酸氢氯吡格雷晶型中的基因毒性杂质对甲苯磺酸甲酯

目的：建立超高效液相色谱-三重四极杆质谱联用法（UHPLC-MS）测定2种硫酸氢氯吡格雷晶型（Ⅰ型和Ⅱ型）中基因毒性杂质对甲苯磺酸甲酯。方法：采用Zorbax Extend C₁₈（2.1 mm×50 mm，1.8 μm）色谱柱，以0.1%甲酸水溶液为流动相A，0.1%甲酸甲醇溶液为流动相B，梯度洗脱，流速为0.4 mL·min^-1，电喷雾（ESI）正离子检测模式，采用多反应监测（MRM）扫描模式，选择m/z 187→91作为定量离子对，m/z 187→155作为定性离子对。结果：对甲苯磺酸甲酯质量浓度在1.0~200 ng·mL^-1范围内线性关系良好（r=0.999 9）；检测限为0.5 ng·mL^-1；定量限为1.0 ng·mL^-1；Ⅰ型晶型和Ⅱ型晶型的加样回收率分别为97.7%和97.9%（n=9），RSD分别为1.5%和1.5%；4℃控温条件下供试品溶液在48 h内稳定性良好（RSD=1.6%）；2种晶型各3批样品中均未检出对甲苯磺酸甲酯。结论：本方法结果可靠，可以用于硫酸氢氯吡格雷原料中对甲苯磺酸甲酯的检测。

Objective: To determine genotoxic impurity methyl p-toluenesulfonate in two kinds of crystal forms(crystal form I and crystal form Ⅱ) of clopidogrel hydrogen sulfate by UHPLC-MS;Methods: The separation was performed on a Zorbax Extend C₁₈ column(2.1 mm×50 mm, 1.8 μm) with the mobile phase consisting of 0.1% formic acid aqueous solution(mobile phase A) and 0.1% formic acid methanol(mobile phase B) by gradient elution at a flow rate of 0.4 mL·min^-1.Multiple reaction monitoring(MRM) was performed on a triple quadrupole mass spectrometer equipped with a ESI source in the positive mode, selecting m/z 187→91 as the quantitative ion pair and m/z 187→155 as qualitative ion pair.Results: The calibration curve was in a good linearity within the range of 1.0-200 ng·mL^-1(r=0.999 9).The limit of detection was 0.5 ng·mL^-1.The limit of quantification was 1.0 ng·mL^-1.The spike recovery of the two crystal forms were 97.7% and 97.9%(n=9), with RSD of 1.5% and 1.5%, respectively.The sample solution was stable under 4℃ within 48 hours(RSD=1.6%).Methyl p-toluenesulfonate was not detected in sample solutions of two crystal forms.Conclusion: The method is accurate and reliable to determine methyl p-toluenesulfonate in clopidogrel hydrogen sulfate substance.