高效液相色谱串联质谱法测定氯沙坦钾中的遗传毒性杂质N-亚硝基-N-甲基-4-氨基丁酸

目的建立了一种高效液相色谱-三重四极杆串联质谱法,对氯沙坦钾原料药中遗传毒性杂质N-亚硝基-N-甲基-4-氨基丁酸(NMBA)进行测定。方法色谱柱为岛津Shim-pack XR-ODSⅡ色谱柱(2.0 mm×150 mm,2.2μm),流动相为0.1%甲酸水溶液(A)和甲醇(B),进行梯度洗脱,流速0.3 mL·min^-1,柱温为40℃,采用ESI离子化-三重四极杆质谱多反应监测(MRM)正离子模式检测,碰撞电压分别为-11,-13和-13 V,碰撞气氩气270 kPa,NMBA的离子对分别为m/z 147.15→117.10,147.15→87.10和147.15→44.10。结果该方法中NMBA在1~100 ng·mL^-1内线性关系良好,日内和日间的保留时间和峰面积的重复性良好(RSD均小于1.10%,n=6和n=18),低、中、高3个浓度的平均回收率在94.40%~98.04%之间。结论本方法简单方便,可快速有效的对氯沙坦钾原料药中NMBA进行限度检查并实现定量分析。

Determination of N-Nitroso-N-Methyl-4-Aminobutyric Acid in Losartan Potassium Using High Performance Liquid Chromatography Triple Quadrupole Mass Spectrometry

OBJECTIVE To establish a method to determine the genotoxic impurity,N-nitroso-N-methyl-4-aminobytyric acid,in losartan potassium using high performance liquid chromatography triple quadrupole mass spectrometry(HPLC-MS/MS).METHODS The method was developed by using Shimadzu Shim-pack XR-ODS II column(2.0 mm×150 mm,2.2μm).Time program was conducted with mobile phase consisting of water(0.1%formic acid,A)and methanol(B).The flow rate was 0.3 mL·min^-1,and the column oven temperature was maintained at 40℃.The samples were ionized by electrospray ionization(ESI)with multi reaction monitoring(MRM)data acquisition mode.The collision energies were-11,-13,and-13 V,CID gas was argon with pressure of 270 k Pa.3 pairs of precursor,and product ions(m/z)of NMBA were 147.15→117.10,147.15→87.10,and 147.15→44.10,respectively.RESULTS The genotoxic impurity NMBA showed linearity between 1 and 100 ng·m L^-1 with correlation coefficient of 0.9999.The intra-day and inter-day repeatability was examined by relative standard deviations(RSDs)of retention time and peak area(RSD<1.10%,n=6 for intra-day repeatability and n=18 for inter-day repeatability).The accuracy was examined by percent recovery at three concentration levels,and the average percent recovery was between 94.40%and 98.04%.CONCLUSION The established LC-MS/MS method is efficient for limit test and quantitation of NMBA in losartan potassium bulk drug.