UPLC-MS/MS法同时测定大鼠血浆中氟西汀和诺氟西汀的含量
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摘要
目的:建立快速测定大鼠血浆中氟西汀和诺氟西汀含量的超高效液相色谱-质谱联用(UPLC-MS/MS)方法并用于大鼠体内的药动学研究。方法:采用乙腈一步沉淀法,以帕罗西汀为内标,色谱柱为Waters Acquity UPLC BEH C_(18)柱(50 mm×2.1 mm,1.7μm),流动相为乙腈和0.1%甲酸,梯度洗脱,流速为0.4 ml·min~(-1),柱温为40℃。质谱采用电喷雾(ESI)正离子模式,选择多反应监测(MRM),氟西汀、诺氟西汀和内标帕罗西汀的离子对分别为m/z 310.80→147.96、m/z 296.06→133.97、m/z 330.19→191.99。结果:大鼠血浆中氟西汀、诺氟西汀和内标在3 min内完全分离。氟西汀和诺氟西汀在0.5～250 ng·ml~(-1)的浓度范围内线性关系良好,r分别为0.999 0和0.999 5,检测下限均为0.5 ng·ml~(-1)。日内和日间精密度的RSD均在15%以内,绝对回收率在80%以上。结论:本方法快速、简便、准确,并可应用于大鼠体内药动学的研究。
Objective:To establish a rapid method for the determination of fluoxetine and norfluoxetine in rat’s plasma by ultrahigh performance liquid chromatography-mass spectrometry(UPLC-MS/MS)and apply in pharmacokinetic study in rats.Methods:A one-step precipitation of acetonitrile was used with paroxetine as the internal standard.A Waters Acquity UPLC BEH C_(18) column(50mm×2.1 mm,1.7μm)was used.The mobile phase was acetonitrile and 0.1%formic acid with gradient elution at a flow rate of 0.4ml·min~(-1)and the column temperature was 40℃.The electrospray ionization(ESI)in positive ion mode with multiple reaction monitoring(MRM)was selected for ionization of the analytes.The major ion transition selected for detection of fluoxetine,norfluoxetine and the internal standard paroxetine was m/z 310.80→147.96,m/z 296.06→133.97 and m/z 330.19→191.99.Results:Fluoxetine,norfluoxetine and the internal standard in rat’s plasma were completely separated within 3 minutes.The linear relationships of fluoxetine and norfluoxetine were good within the concentration range of 0.5-250 ng·ml~(-1).The correlation coefficient was 0.999 0 and 0.999 5,respectively.The detection limit was 0.5 ng·ml~(-1).The RSDs of intra-and inter-day precisions were less than 15% and the recovery was above 80%.Conclusion:The method is rapid,simple and accurate,which can be successfully applied in the pharmacokinetic study in rats.
Objective:To establish a rapid method for the determination of fluoxetine and norfluoxetine in rat’s plasma by ultrahigh performance liquid chromatography-mass spectrometry(UPLC-MS/MS)and apply in pharmacokinetic study in rats.Methods:A one-step precipitation of acetonitrile was used with paroxetine as the internal standard.A Waters Acquity UPLC BEH C_(18) column(50mm×2.1 mm,1.7μm)was used.The mobile phase was acetonitrile and 0.1%formic acid with gradient elution at a flow rate of 0.4ml·min~(-1)and the column temperature was 40℃.The electrospray ionization(ESI)in positive ion mode with multiple reaction monitoring(MRM)was selected for ionization of the analytes.The major ion transition selected for detection of fluoxetine,norfluoxetine and the internal standard paroxetine was m/z 310.80→147.96,m/z 296.06→133.97 and m/z 330.19→191.99.Results:Fluoxetine,norfluoxetine and the internal standard in rat’s plasma were completely separated within 3 minutes.The linear relationships of fluoxetine and norfluoxetine were good within the concentration range of 0.5-250 ng·ml~(-1).The correlation coefficient was 0.999 0 and 0.999 5,respectively.The detection limit was 0.5 ng·ml~(-1).The RSDs of intra-and inter-day precisions were less than 15% and the recovery was above 80%.Conclusion:The method is rapid,simple and accurate,which can be successfully applied in the pharmacokinetic study in rats.
引文
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8马国静,郑国艳,赵丛俏,等.HPLC-MS/MS法同时测定大鼠血浆中芬太尼、氟西汀及其代谢物的质量浓度及药动学研究[J].沈阳药科大学学报,2018,35(5):390-398
9 Ni XJ,Wang ZZ,Shang DW,et al.Simultaneous analysis of olanzapine,fluoxetine,and norfluoxetine in human plasma using liquid chromatography-mass spectrometry and its application to a pharmacokinetic study[J].J Chromatogr B Analyt Technol Biomed Life Sci,2018,1092:506-514
10 Magalhaes P,Alves G,Llerena A,et al.Therapeutic Drug Monitoring of Fluoxetine,Norfluoxetine and Paroxetine:A New Tool Based on Microextraction by Packed Sorbent Coupled to Liquid Chromatography[J].J Anal Toxicol,2017,41(7):631-638
11 da Silva ACC,Raasch JR,Vargas TG,et al.Simultaneous determination of fluoxetine and norfluoxetine in dried blood spots using highperformance liquid chromatography-tandem mass spectrometry[J].Clin Biochem,2018,52:85-93
2 Macias-Cortes Edel C,Llanes-Gonzalez L,Aguilar-Faisal L,et al.Individualized homeopathic treatment and fluoxetine for moderate to severe depression in peri-and postmenopausal women(HOMDEP-MENOP study):a randomized,double-dummy,double-blind,placebo-controlled trial[J].Plo S One,2015,10(3):e0118440
3 Zhang K,Cao L,Zhu W,et al.Association between the efficacy of fluoxetine treatment in obsessive-compulsive disorder patients and SLC1A1 in a Han Chinese population[J].Psychiatry Res,2015,229(1-2):631-632
4 Tanoshima R,Bournissen FG,Tanigawara Y,et al.Population PK-modelling and simulation based on fluoxetine and norfluoxetine concentrations in milk:a milk concentration-based prediction model[J].Br J Clin Pharmacol,2014,78(4):918-928
5 Sager JE,Lutz JD,Foti RS,et al.Fluoxetine-and norfluoxetine-mediated complex drug-drug interactions:in vitro to in vivo correlation of effects on CYP2D6,CYP2C19,and CYP3A4[J].Clin Pharmacol T-her,2014,95(6):653-662
6 Ji Y,Chen S,Zhao L,et al.In vitro assessment of 39 CYP2C9 variants found in the Chinese population on the metabolism of the model substrate fluoxetine and a summary of their effects on other substrates[J].J Clin Pharm Ther,2015,40(3):320-327
7王欣晨,许杜娟,夏泉,等.互为内标法测定患者血浆中氟西汀和西酞普兰的浓度[J].中国药师,2012,15(10):1431-1434
8马国静,郑国艳,赵丛俏,等.HPLC-MS/MS法同时测定大鼠血浆中芬太尼、氟西汀及其代谢物的质量浓度及药动学研究[J].沈阳药科大学学报,2018,35(5):390-398
9 Ni XJ,Wang ZZ,Shang DW,et al.Simultaneous analysis of olanzapine,fluoxetine,and norfluoxetine in human plasma using liquid chromatography-mass spectrometry and its application to a pharmacokinetic study[J].J Chromatogr B Analyt Technol Biomed Life Sci,2018,1092:506-514
10 Magalhaes P,Alves G,Llerena A,et al.Therapeutic Drug Monitoring of Fluoxetine,Norfluoxetine and Paroxetine:A New Tool Based on Microextraction by Packed Sorbent Coupled to Liquid Chromatography[J].J Anal Toxicol,2017,41(7):631-638
11 da Silva ACC,Raasch JR,Vargas TG,et al.Simultaneous determination of fluoxetine and norfluoxetine in dried blood spots using highperformance liquid chromatography-tandem mass spectrometry[J].Clin Biochem,2018,52:85-93