UPLC-APCI MS/MS法检测克拉霉素中3个N-亚硝胺类化合物

Determination of N-nitrosamines(NIPEA,NDIPA,NDBA)in clarithromycin by UPLC-APCI MS/MS

目的:建立超高液相色谱-大气压化学电离源三重四极杆质谱法(UPLC-APCI MS/MS)检测克拉霉素中3个N-亚硝胺类化合物[N-亚硝基乙基异丙基胺(NIPEA)、N-亚硝基二异丙胺(NDIPA)和N-亚硝基二丁胺(NDBA)。方法:采用ACQUITY UPLC HSS T3色谱柱(100 mm×2.1 mm,1.8μm),以甲醇-0.1%甲酸水溶液为流动相,梯度洗脱,流速0.4 mL·min^(-1);采用大气压化学电离(APCI),多反应监测(multiple reaction monitoring,MRM),正离子扫描模式。结果:测得NIPEA、NDIPA质量浓度在18~120μg·L^(-1)内呈现良好的线性关系(r=0.999),NDBA质量浓度在7.8~52μg·L^(-1)内呈现良好的线性关系(r=0.9991),重复性试验的RSD为2%~3%,方法回收率为88%~109%,NIPEA、NDIPA方法定量限(S/N不低于10)为9.0μg·L^(-1),NDBA方法定量限(S/N不低于10)为3.9μg·L^(-1),溶液在室温放置24 h内稳定。对2家国内生产企业各3批克拉霉素样品进行检测,NIPEA、NDIPA、NDBA均低于检测限。结论:本试验建立了克拉霉素中N-亚硝胺类化合物NIPEA、NDIPA、NDBA的检测方法,具有灵敏度高,测定结果准确,专属性强等诸多优点,可应用于临床治疗药物的监测。

Objective:To establish an UPLC-APCI MS/MS method for the determination of three N-nitrosamines(NIPEA、NDIPA、NDBA)in clarithromycin.Methods:An ACQUITY UPLC HSS T3(100 mm×2.1 mm,1.8μm)was adopted,the mobile phase consisted of methanol-0.1%formic acid solution with gradient elution,at a flow rate of 0.4 mL·min^(-1);atmospheric-pressure chemical ionization(APCI),multiple reaction monitoring(MRM),positive ion scanning mode were used.Results:The measured mass concentrations of NIPEA and NDIPA had a good linearity in the range of 18-120μg·L^(-1)(r=0.999),and the mass concentration of NDBA had a good linearity in the range of 7.8-52μg·L^(-1)(r=0.9991).The RSD of all repeatability tests were 2%-3%,the recovery rates were 88%-109%.The LOQ(S/N was about 10)of the methods for NIPEA andNDIPA were 9.0μg·L^(-1)and that for NDBA was 3.9μg·L^(-1).The solution was stable at room temperature in 24 h.Three batches of clarithromycin samples from two domestic manufacturers were tested,and NIPEA,NDIPA and NDBA were below LOD.Conclusion:This study established a method for the determination of N-nitrosamines NIPEA,NDIPA and NDBA in clarithromycin with the advantages of high sensitivity,accurate determination results,strong specificity and so on.It can be applied to the monitoring of clinical therapeutic drugs.