GC-MS/MS测定替米沙坦片中10种亚硝胺类基因毒性杂质

Determination of Ten Nitrosamine Genotoxic Impurities in Telmisartan Tablets by GC-MS/MS

目的 建立GC-MS/MS同时测定替米沙坦片中N-亚硝基二甲胺(NDMA)、N-亚硝基甲乙胺(NMEA)、N-亚硝基二乙胺(NDEA)、N-亚硝基-N-乙基异丙胺(NEIPA)、N-亚硝基二异丙胺(NDIPA)、N-亚硝基二正丙胺(NDPA)、N-亚硝基二正丁胺(NDBA)、N-亚硝基哌啶(NPIP)、N-亚硝基吡咯烷(NPYR)和N-亚硝基吗啉(NMOR)10种亚硝胺类基因毒性杂质的含量。方法样品经甲醇提取,经Agilent VF-WAXms(30 m×0.25 mm,1μm)毛细管气相色谱柱分离,采用多反应离子监测(MRM)模式进行定量分析。结果 NDMA、NMEA、NDEA、NEIPA、NDIPA、NDPA、NDBA和NPIP在0.2~50 ng·mL^(–1)线性关系良好,相关系数均为1.000 0;检测限均为0.05 ng·mL^(–1),定量限均为0.2 ng·mL^(–1),平均回收率分别为103%(RSD=9.2%,n=9),108%(RSD=6.1%,n=9),107%(RSD=5.3%,n=9),106%(RSD=3.9%,n=9),102%(RSD=5.0%,n=9),99%(RSD=6.9%,n=9),97%(RSD=8.6%,n=9),101%(RSD=4.4%,n=9)。NPYR和NMOR在1.0~50 ng·mL^(–1)内线性关系良好,相关系数均为1.000 0;检测限均为0.4 ng·mL^(–1),定量限均为1.0 ng·mL^(–1),平均回收率分别为95%(RSD=6.4%,n=9),103%(RSD=6.1%,n=9)。所收集的254批样品中,12批次检出NDMA,其余9个杂质在254批样品中均未检出。结论 该方法操作简便、灵敏度高,专属性强,适用于替米沙坦片或其他制剂中NDMA、NMEA、NDEA、NEIPA、NDIPA、NDPA、NDBA、NPIP、NPYR和NMOR 10种基因毒性杂质的检测。

OBJECTIVE To establish a GC-MS/MS method for simultaneous determination of ten nitrosamine genotoxic impurities in telmisartan tablets,including N-nitrosodimethylamine(NDMA),N-nitrosomethylethylamine(NMEA),N-nitrosodiethylamine(NDEA),N-nitroso-N-ethylisopropylamine(NEIPA),N-nitrosodiisopropylamine(NDIPA),Nnitrosodipropylamine(NDPA),N-nitrosodibutylamine(NDBA),N-nitrosopiperidine(NPIP),N-nitrosopyrrolidine(NPYR) and N-nitrosomorphpline(NMOR).METHODS The sample was extracted with methanol.The sample was separated by an Agilent VF-WAXms(30 m×0.25 mm,1 μm) capillary gas chromatography column,and the multiple reactive ion monitoring(MRM) was selected for the quantitative detection.RESULTS NDMA,NMEA,NDEA,NEIPA,NDIPA,NDPA,NDBA and NPIP displayed satisfactory linearity(r=1.000 0) over the concentration range of 0.2-50 ng·mL^(–1).The limits of detection and the limits of quantification were 0.05 ng·mL^(–1) and 0.2 ng·mL^(–1).The average recoveries were 103%(RSD=9.2%,n=9),108%(RSD=6.1%,n=9),107%(RSD=5.3%,n=9),106%(RSD=3.9%,n=9),102%(RSD=5.0%,n=9),99%(RSD=6.9%,n=9),97%(RSD=8.6%,n=9),101%(RSD=4.4%,n=9).NPYR and NMOR displayed satisfactory linearity(r=1.000 0) over the concentration range of 1.0-50 ng·mL^(–1).The limits of detection and the limits of quantification were 0.4 ng·mL^(–1) and 1.0 ng·mL^(–1).The average recoveries were 95%(RSD=6.4%,n=9),103%(RSD=6.1%,n=9).The 254 batches of telmisartan tablets were analyzed by this method,and NDMA was detected in 12 batches.The remaining nine impurities were not detected in 254 batches of sample.CONCLUSION The method is simple,sensitive with strong specificity.It is suitable for the detection of NDMA,NMEA,NDEA,NEIPA,NDIPA,NDPA,NDBA,NPIP,NPYR and NMOR in telmisartan tablets and other preparations.