HPLC-MS/MS法测定注射用盐酸头孢甲肟中7种N-亚硝胺类杂质

Determination of 7 N-nitrosamine impurities in cefmenoxime hydrochloride for injection by HPLC-MS/MS

目的建立高效液相色谱-串联质谱法(HPLC-MS/MS)同时测定注射用盐酸头孢甲肟中7种N-亚硝胺类基因毒性杂质(N-亚硝基二甲胺、N-亚硝基二乙胺、N-亚硝基二乙醇胺、N-亚硝基-N-甲基苯胺、N-亚硝基-N-乙基苯胺、N-亚硝基二异丙胺、N-亚硝基二正丁胺)。方法色谱柱为ACE EXCEL 3 C18-AR(3μm,100 mm×4.6 mm),流动相为0.1%甲酸甲醇溶液(A)-0.1%甲酸(B),梯度洗脱,流速0.6 mL·min^(-1),采用APCI离子源,以正离子、多重反应监测(MRM)模式进行检测。结果7种N-亚硝胺类基因毒性杂质在1~20ng·mL^(-1)与峰面积线性关系良好。回收率在78.9%~89.6%,RSD均小于6.1%。检测限低于1.5ng·g^(-1)。样品中各杂质均未检出。结论该方法灵敏度高、专属性强,可用于注射用盐酸头孢甲肟中7种N-亚硝胺类基因毒性杂质的质量控制。

Objective To determine 7 N-nitrosamine impurities(N-nitrosodimethylamine,N-nitrosodiethylamine,N-nitrosodiethanolamine,N-nitroso-N-methylphenylamine,N-nitroso-N-ethylphenylamine,N-nitrosodiipropylamine,and N-nitrosodibutylamine)in cefmenoxime hydrochloride for injection by HPLC-MS/MS.Methods The separation was performed on an ACE EXCEL 3 C18-AR(3μm,100 mm×4.6 mm)column with the mobile phase 0.1%formic acid methanol solution(A)-0.1%formic acid solution(B)by gradient elution at a flow rate of 0.6 mL·min^(-1).Multiple reaction monitoring was performed on a triple quadripole mass spectrometer equipped with an APCI source in the positive mode.Results The calibration curves were in good linearty at 1~20 ng·mL^(-1) for 7 N-nitrosamine impurities.The recoveries were within 78.9%~89.6%with RSD under 6.1%.The limit of detection was less than 1.5 ng•g^(-1).No impurities were detected in the samples.Conclusion The HPLC-MS/MS method is accurate,highly sensitive,strong specific,and can be used for the quality control of 7 N-nitrosamine genotoxic impurities in cefmenoxime hydrochloride for injection.