LC-MS/MS测定酒石酸溴莫尼定中苯胺类基因毒性杂质

Determination of Anililines Genotoxic Impurity in Brimonidine Tartrate by LC-MS/MS

关键词:  酒石酸溴莫尼定|苯胺类基因毒性杂质|液相色谱-质谱联用|质量控制

Key words:  brimonidine tartrate|anililines genotoxic impurity|LC-MS/MS|quality control

摘要:

目的 建立LC-MS/MS方法测定酒石酸溴莫尼定中苯胺类基因毒性杂质（6-氨基喹喔啉、4-硝基邻苯二胺、6-氨基-5-溴喹喔啉）的含量。方法 采用色谱柱ZORBAX Eclipse Plus C₁₈（2.1 mm×50 mm，1.8 μm），流动相为0.1%乙酸水溶液（A）-乙腈（B），梯度洗脱，流速0.35 mL·min^-1，柱温为35℃；采用Shimadzu LC30 UPLC-AB Science 4000+三重四极杆液质联用仪和多反应离子监测模式二级质谱MS/MS（ESI）源检测，正离子模式采集数据。结果 6-氨基喹喔啉、4-硝基邻苯二胺、6-氨基-5-溴喹喔啉分别在9.98~199.50，9.94~198.74，10.03~200.55 ng·mL^-1内线性关系良好；低、中、高浓度（50，100，150 ng·mL^-1）回收率为94.8%~110.8%，RSD均<4.2%；溶液在12 h内稳定性良好。结论 本方法操作简便，结果可靠，可以用于酒石酸溴莫尼定中苯胺类基因毒性杂质（6-氨基喹喔啉、4-硝基邻苯二胺、6-氨基-5-溴喹喔啉）的控制。

Abstract:

OBJECTIVE To establish LC-MS/MS method for the determination of anililines genotoxic impurities (6-aminoquinoxaline, 4-nitro-o-phenylenediamine, and 6-amino-5-bromoquinoxaline) in brimonidine tartrate. METHODS The method was achieved on a ZORBAX Eclipse Plus C₁₈ column(2.1 mm×50 mm, 1.8 μm) utilizing a mobile phase of 0.1% acetic acid(A)-acetonitrile(B) with gradient elution at the flow rate of 0.35 mL·min^-1. The temperature of column was set at 35℃. The Shimadzu LC30 UPLC-AB Science 4000+ LC-MS was used to detect(ESI source) in multiple reaction monitoring positive mode. RESULTS The calibration curves of 6-aminoquinoxaline, 4-nitro-o-phenylenediamine, 6-amino-5-bromoquinoxaline were linear in 9.98-199.50, 9.94-198.74, 10.03-200.55 ng·mL^-1. The average recoveries of the control samples at low, middle and high concentrations(50, 100, 150 ng·mL^-1) were 94.8%-110.8%, all RSDs were <4.2%, solution stability was good within 12 h. CONCLUSION The method is convenient to operate and has reliable results for the determination of anililines genotoxic impurities of 6-aminoquinoxaline, 4-nitro-o-phenylenediamine, 6-amino-5-bromoquinoxaline in bromnidine tartrate.