超高效液相色谱-静电场轨道阱高分辨质谱法测定法莫替丁及其制剂中的痕量N-亚硝基二甲胺

Determination of Trace N-nitrosodimethylamine in Famotidine and Its Preparations by Ultra-high Performance Liquid Chromatography-Orbitrap High Resolution Mass Spectrometry

建立了测定法莫替丁及其制剂中N-亚硝基二甲胺(NDMA)含量的超高效液相色谱-静电场轨道阱高分辨质谱法(UHPLC-Orbitrap HRMS)。样品以甲醇作为提取溶剂,经涡旋混匀、恒温振荡、高速离心、微孔过滤后进行液相色谱-质谱(LC-MS)分析。采用ACE EXCEL 3 C18-AR(150 mm×4.6 mm,3μm)色谱柱,以0.1%甲酸水溶液和0.1%甲酸乙腈为流动相梯度洗脱分离,流速为0.50 mL/min,柱温为30℃,自动进样器温度为4℃,设置六通阀切换保护质谱系统。质谱分析采用ESI离子源,正离子平行反应监测(PRM)扫描模式,外标法定量。NDMA在1.00~100.00 ng/mL范围内线性良好,相关系数(r)为0.9997,检出限和定量下限分别为0.20 ng/mL和1.00 ng/mL,在法莫替丁及其制剂中的平均回收率为98.5%~108%,相对标准偏差(RSD)为2.3%~6.7%。将该法用于47批供试品中NDMA的测定,在1批法莫替丁原料和2批制剂中检出NDMA,其含量超过限度规定。该方法灵敏、准确、操作简便,可用于法莫替丁及其制剂中NDMA的测定。

An ultra-high performance liquid chromatography-orbitrap high resolution mass spectrometry(UHPLC-Orbitrap HRMS)was used for the determination of N-nitrosodimethylamine(NDMA)in famotidine and its preparations.The samples were extracted with methanol,followed by vortex mixing,constant temperature shaking,high speed centrifugation and microfiltration,and finally analyzed by liquid chromatography-mass spectrometry(LC-MS).The chromatographic separation was performed on an ACE EXCEL 3 C18-AR(150 mm×4.6 mm,3μm)column at 30℃by gradient elution,with water and acetonitrile both contained 0.1%formic acid as mobile phases at a flow rate of 0.50 mL/min.The temperature of the autosampler was set at 4℃,and the mass spectrometer was protected with a valve switching apparatus.The MS analysis was performed with ESI ion source in positive ion parallel reaction monitoring(PRM)scanning mode,and the quantitation was achieved by the external standard method.Results showed that there was a good linear relationships for NDMA in the concentration range of 1.00-100.00 ng/mL with correlation coefficient(r)of 0.9997.The limit of detection and limit of quantitation were 0.20 ng/mL and 1.00 ng/mL,respectively.The mean recoveries of NDMA from spiked famotidine and its preparations ranged from 98.5%to 108%with relative standard deviations(RSD)of 2.3%-6.7%.The method was then applied to the determination of NDMA in famotidine and its preparations.Among 47 batches of samples,NDMA was detected in 1 batch of famotidine and 2 batches of famotidine preparations,which contents exceeded the limit requirement.This method is sensitive,accurate and easy to operate,and could be used for the determination of NDMA in famotidine and its preparations.