高效液相色谱-高分辨质谱法测定盐酸替罗非班中偶氮类遗传毒性杂质  

Determination of azo genotoxic impurities in tirofiban hydrochloride by high performance liquid chromatography-high resolution mass spectrometry

作　　者：张申奥 左呈露 胡林 ZHANG Shen-ao;ZUO Cheng-lu;HU Lin(School of Pharmacy,Anhui University of Chinese Medicine,Hefei 230013;Yangtze Delta Advanced Research Institute,Nantong Jiangsu 226133;Wisdom Pharmaceutical Co.,Ltd.,Nantong Jiangsu 226123)

机构地区：[1]安徽中医药大学药学院,合肥230013 [2]长三角药物高等研究院,江苏南通226133 [3]江苏慧聚药业股份有限公司,江苏南通226123

出　　处：《中南药学》2025年第3期730-735,共6页Central South Pharmacy

摘　　要：目的采用高效液相色谱-高分辨质谱(HPLC-HRMS)法测定盐酸替罗非班中偶氮二甲酸二乙酯的含量。方法在一定浓度的水合肼溶液中,偶氮二甲酸二乙酯与水合肼反应合成1,2-肼二羧酸二乙酯,采用Phenomenex Luna C18(2)100Å(2 mm×50 mm,3μm)色谱柱,以乙腈(A)-0.1%的甲酸溶液(B)为流动相,梯度洗脱,流速0.8 mL·min^(-1),柱温40℃,进样体积2μL,离子源为电喷雾离子源(ESI),采用正离子模式下单离子监测模式(SIM)监测。结果衍生后的偶氮二甲酸二乙酯在198.0396~1485.2970 ng·mL^(-1)与峰面积呈良好的线性关系(r=0.9980);定量限为9.7058 ng·mL^(-1),检测限为1.9412 ng·mL^(-1);低、中、高浓度的加样回收率均在95.5%~106.8%,RSD≤3.6%。6批盐酸替罗非班原料药中均未检出偶氮二甲酸二乙酯。结论所建立的方法操作简单,灵敏度高,测试结果准确,可用于盐酸替罗非班中偶氮二甲酸二乙酯总残留量的检测。Objective To determine diethyl azodicarboxylate in tirofiban hydrochloride by high performance liquid chromatography-high resolution mass spectrometry(HPLC-HRMS).Methods Diethyl 1,2-hydrazinedicar-boxylate was synthesized in a specific concentration of hydrazine hydrate solution,and diethyl azodicarboxylate reacted with hydrazine hydrate.The derivatization-HPLCHRMS was equipped with a Phenomenex Luna C18(2)100Åcolumn(2 mm×50 mm,3μm).The mobile phase consisted of acetonitrile(A)-0.1%formic acid solution(B)with gradient elution at 0.8 mL·min^(-1).The column temperature was 40℃,and injection volume was 2μL.Electrospray ionization(ESI)was used to detect the positive ions by single ion monitor(SIM).Results The derived genotoxic impurity had a good linearity at 198.0396~1485.2970 ng·mL^(-1)(r=0.9980).The limit of quantitative was 9.7058 ng·mL^(-1) and the limit of detection was 1.9412 ng·mL^(-1).The recovery of low,medium and high concentrations ranged 95.5%~106.8%,with RSD≤3.6%.Diethyl azodicarboxylate was not detected in the 6 batches of tirofiban hydrochloride bulk drug samples.Conclusion The method is easily operable,with high sensitivity and accuracy.It can be used to detect of diethyl azodicarboxylate in tirofiban hydrochloride.

关 键 词：盐酸替罗非班 高效液相色谱-高分辨质谱法 MITSUNOBU反应 偶氮二甲酸二乙酯 柱前衍生 遗传毒性杂质 

分 类 号：R927.2[医药卫生—药学]