摘要
目的通过合成基因毒性杂质N-亚硝基氢氯噻嗪(NO-HZCT),建立超高效液相色谱-串联质谱法(UHPLC-MS/MS)测定依那普利氢氯噻嗪制剂中N-亚硝基氢氯噻嗪。方法参考文献方法合成NO-HZCT,采用高分辨质谱对其相对分子量和结构进行确定;采用Agilent Eclipse Plus C_(18)RRHD(3.0 mm×150 mm,1.8μm)色谱柱,以10 mmol·L^(-1)甲酸铵-0.1%甲酸的水溶液作为流动相A,以0.1%甲酸的乙腈溶液作为流动相B,梯度洗脱,体积流量0.6 mL·min^(-1);采用ESI离子源正离子扫描,多反应监测(MRM)模式下,对NO-HZCT进行定量检测。结果NO-HZCT质量浓度在0.51~50.67 ng·mL^(-1)范围内具有良好的线性关系,相关系数(r)为0.9997;低、中、高3个浓度的加样回收率(n=3)分别为93.10%(RSD 3.7%)、104.30%(RSD 1.0%)和106.48%(RSD 1.8%);检测限和定量限分别为0.08 ng·mL^(-1)和0.27 ng·mL^(-1),3批样品中均检测出NO-HZCT。结论该方法灵敏度高、专属性强,可准确地对依那普利氢氯噻嗪制剂中遗传毒性杂质NO-HZCT进行定量检测,可为依那普利氢氯噻嗪制剂的质量控制提供参考,保障药品质量安全。
Objective To establish an UHPLC-MS/MS method for the determination of N-nitroso hydrochlorothiazide in enalapril hydrochlorothiazide preparations by synthesizing the genotoxic impurity N-nitroso hydrochlorothiazide(NO-HZCT).Methods NO-HZCT was synthesized by reference method,its molecular weight and structure were confirmed by high resolution mass spectrometry;The separation was performed on an Agilent Eclipse Plus C 18 RRHD(3.0 mm×150 mm,1.8μm)column with the mobile phase consisting of 10 mmol·L^(-1) ammonium formate-0.1%formic acid aqueous solution(mobile phase A)and 0.1%formic acid solution(mobile phase B)by gradient elution at a flow rate 0.6 mL·min^(-1);Multiple reaction monitoring(MRM)was performed on a triple quadruple mass spectrometer with a ESI source in positive mode to quantitative detection of NO-HZCT.Results The calibration curve was linear for NO-HZCT in the range 0.51~50.67 ng·mL^(-1),Coefficient of correlation(r)was 0.9997;The recoveries(n=3)of low,middle and high adding concentrations were 93.10%(RSD 3.7%),104.30%(RSD 1.0%)and 106.48%(RSD 1.8%),respectively;The LOD and LOQ were 0.08 ng·mL^(-1) and 0.027 ng·mL^(-1),respectively.NO-HZCT was detected in three batches of samples.Conclusion The method was highly sensitive,accurate,and can accurately detect the genotoxic impurity NO-HZCT in enalapril hydrochlorothiazide preparations,which can provide a reference for the quality control of enalapril hydrochlorothiazide preparations and ensure the drug safety.
作者
袁松
张才煜
庾莉菊
陈华
张庆生
YUAN Song;ZHANG Caiyu;YU Liju;CHEN Hua;ZHANG Qingsheng(NMPA Key Laboratory for Quality Research and Evaluation of Chemical Drugs,National Institute for Food and Drug Control,Beijing 102629,China)
出处
《药学研究》
CAS
2023年第8期579-583,共5页
Journal of Pharmaceutical Research
基金
中国食品药品检定研究院关键技术研究基金(No.GJJS-2022-4-1)。
