UPLC-MS/MS法同时检测大鼠血浆伊马替尼及其代谢物
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摘要
目的建立一种同时快速测定大鼠血浆中伊马替尼及其活性代谢产物N-去甲基伊马替尼浓度的超高效液相色谱串联质谱(UPLC-MS/MS)法。方法用乙腈溶剂沉淀血浆蛋白的前处理方法,应用三重四级杆液质联用仪,色谱柱为ACQUITY UPLC BEH C_(18)色谱柱(2. 1 mm×50 mm,1. 7μm);流动相为乙腈-水(含0. 1%甲酸),梯度洗脱,流速为0. 4 ml/min,柱温40℃,以卡马西平为内标;质谱条件:ESI离子源,正离子模式。结果伊马替尼的出峰时间为0. 38 min,线性范围为20～10 000 ng/ml (r~2=0. 999 2),最低定量限为20 ng/ml,回收率为87. 67%～92. 61%,日内、日间精密度RSD均<15%。N-去甲基伊马替尼的出峰时间为0. 36 min,线性范围为1～1 000 ng/ml (r~2=0. 999 5),最低定量限为1 ng/ml,回收率为87. 26%～98. 89%,日内、日间精密度RSD均<15%。结论该法操作简便快速,专属性高,重复性好,灵敏度高,适用于大鼠血浆中伊马替尼及其活性代谢产物N-去甲基伊马替尼的浓度检测。
Objective To construct a UPLC-MS/MS method for determining the concentration of imatinib and its active metabolite N-desmethyl imatinib in rat' s plasma. Methods The plasma was precipitated by acetonitrile. An ACQUITY UPLC BEH C_(18)( 2.1 mm × 50 mm,1. 7 μm) column was used as the stationary phase on Waters XEVO TQD. The temperature of column was 40 ℃ and the mobile phase consisted of acetonitrile and water( containing 0. 1%formic acid) at gradient elution. The flowrate of the mobile phase was 0. 4 ml/min. The internal standard was carbamazepine. Results The peak time of imatinib was 0. 38 min. Excellent linear calibration curves of imatinib were obtained in the concentration range of 20 ～ 10 000 ng/ml( r~2= 0. 999 2). The lower limit of quantification of imatinib was 20 ng/ml while the average recovery was 87. 67% ～ 92. 61%. The intra-and inter-day RSD both was below15%.The peak time of N-desmethyl imatinib was 0. 36 min. Excellent linear calibration curves of N-desmethyl imatinib were obtained in the concentration range of 1 ～ 1 000 ng/ml( r~2= 0. 999 5). The lower limit of quantification of N-desmethyl imatinib was 1 ng/ml while the average recovery was 87. 26% ～ 98. 89%. The intra-and inter-day RSD both was below 15%. Conclusion The UPLC-MS/MS method is simple,quick and accurate with high sensitivity,and it is suitable for the determination of imatinib and its active metabolite N-desmethyl imatinib in rat's plasma.
Objective To construct a UPLC-MS/MS method for determining the concentration of imatinib and its active metabolite N-desmethyl imatinib in rat' s plasma. Methods The plasma was precipitated by acetonitrile. An ACQUITY UPLC BEH C_(18)( 2.1 mm × 50 mm,1. 7 μm) column was used as the stationary phase on Waters XEVO TQD. The temperature of column was 40 ℃ and the mobile phase consisted of acetonitrile and water( containing 0. 1%formic acid) at gradient elution. The flowrate of the mobile phase was 0. 4 ml/min. The internal standard was carbamazepine. Results The peak time of imatinib was 0. 38 min. Excellent linear calibration curves of imatinib were obtained in the concentration range of 20 ～ 10 000 ng/ml( r~2= 0. 999 2). The lower limit of quantification of imatinib was 20 ng/ml while the average recovery was 87. 67% ～ 92. 61%. The intra-and inter-day RSD both was below15%.The peak time of N-desmethyl imatinib was 0. 36 min. Excellent linear calibration curves of N-desmethyl imatinib were obtained in the concentration range of 1 ～ 1 000 ng/ml( r~2= 0. 999 5). The lower limit of quantification of N-desmethyl imatinib was 1 ng/ml while the average recovery was 87. 26% ～ 98. 89%. The intra-and inter-day RSD both was below 15%. Conclusion The UPLC-MS/MS method is simple,quick and accurate with high sensitivity,and it is suitable for the determination of imatinib and its active metabolite N-desmethyl imatinib in rat's plasma.
引文
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[2]丁倩倩,陈勤奋.甲磺酸伊马替尼的临床应用[J].上海医药,2013,34(3):5-9.
[3]王晨明,林高通,王哲,等.HPLC检测大鼠血浆中伊马替尼的浓度及其药动学研究[J].中国药学杂志,2013,48(4):293-296.
[4]许勇,刘明达.大鼠血浆内甲磺酸伊马替尼浓度的测定及其药动学研究[J].中南药学,2013,11(11):818-820.
[5]王军,付翠香,吴伟明,等.高效液相色谱法测定大鼠血浆中伊马替尼浓度及其药动学[J].中国新药与临床杂志,2014,33(3):200-203.
[6]罗兴献,黄琳,李泰锋,等.反相高效液相色谱法同时测定伊马替尼及伏立康唑血药浓度[J].中国医院药学杂志,2018,38(1):36-40.
[7]孔宪诚,黄建平,阎良.LC-MS/MS法测定伊马替尼及代谢物在大鼠体内的药动学参数[J].中南药学,2015,13(10):1037-1040.
[8]De Francia S,D'Avolio A,De Martino F,et al.New HPLC-M S method for the simultaneous quantification of the antileukemia drugs imatinib,dasatinib,and nilotinib in human plasma[J].J Chromatogr B Analyt Technol Biomed Life Sci,2009,877(18-19):1721-1726.
[9]孙乃萍,李健,高静,等.胃肠间质瘤患者中Imatinib血药浓度与治疗疗效的相关性[J].基础医学与临床,2010,30(12):1329-1333.
[10]韩勇,周红,张鹏,等.高效液相色谱串联质谱法同时检测人血浆伊马替尼及代谢物[J].中国医院药学杂志,2017,37(21):2122-2126.