超高效液相色谱串联质谱法测定缬沙坦制剂中的2个亚硝胺类遗传毒性杂质

目的 建立超高效液相色谱串联质谱法,同时测定缬沙坦胶囊和分散片中N-亚硝基二甲胺(NDMA)和N-亚硝基二乙胺(NDEA)2个亚硝胺类遗传毒性杂质的残留量。方法 采用安捷伦Zorbax Eclipse Plus C₁₈色谱柱(3.0 mm×150 mm,1.8 μm),以0.1%甲酸水溶液为流动相A,甲醇为流动相B,进行梯度洗脱,流速为0.5 mL·min^-1。质谱采用大气压化学电离(APCI)离子源,多反应监测(MRM)正离子检测模式进行定量分析。结果 NDMA和NDEA线性范围、灵敏度、精密度、准确度、专属性均满足分析要求。经检验,102批缬沙坦胶囊和52批缬沙坦分散片中,NDMA和NDEA含量均不超过定量限浓度,均符合规定。结论 本方法操作简便、快速、准确、灵敏度高、专属性好,适用于缬沙坦胶囊和分散片中NDMA和NDEA的限度检查和定量检测。

Ultra-High Performance Liquid Chromatography Tandem Mass Spectrometry Method for the Determination of Two Nitrosamine Genotoxic Impurities in Valsartan Formulations

OBJECTIVE To establish a method for simultaneous determination of two nitrosamine genotoxic impurities, N-nitrosodimethylamine(NMDA) and N-nitrosodiethylamine(NDEA), in valsartan formulations using ultra-high performance liquid chromatography triple quadrupole mass spectrometry(UHPLC-MS/MS). METHODS The separation was performed on an Agilent Zorbax Eclipse Plus C₁₈ column(3.0 mm×150 mm, 1.8 μm) with mobile phase consisting of 0.1% formic acid aqueous solution(A) and methanol(B) by gradient elution at a flow rate of 0.5 mL·min^-1, and the column oven temperature was maintained at 40 ℃. Multiple reaction monitoring(MRM) was conducted in positive ion mode using atmospheric pressure chemical ionization(APCI) interface. The optimal voltages, currents, and gas flows were chosen to provide the best sensitivity and specificity for each MRM ion pair. Mass transitions m/z 75.0→58.1 and m/z 75.0→43.0 were set as quantitative and qualitative ion pairs for NDMA, and mass transitions m/z 103.0→75.0 and m/z 103.0→47.0 were set as quantitative and qualitative ion pairs for NDEA. RESULTS The calibration curves had good linearity within the ranges of 0.3-50 ng·mL^-1 and 0.06-10 ng·mL^-1 for NDMA and NDEA, respectively. The limits of quantitation(LOQ) of NDMA and NDEA were 0.03 μg·g^-1 and 0.006 μg·g^-1, respectively. The recovery of NDMA was within the range of 92.3%-111.7%(RSD<5.0%), and the recovery of NDEA was between 90.6% and 106.0%(RSD<5.0%). The specificity and precision of the method were also validated. One hundred and two batches of valsartan capsules and 52 batches of valsartan dispersible tablets were analyzed by this method, and the amounts of NDMA and NDEA were all no more than the LOQ values. Therefore, all the samples met the limit requirement. CONCLUSION The established method is simple, rapid, accurate, sensitive and specific for limit test and quantitative analysis of NDMA and NDEA in valsartan formulations.