UPLC-MS/MS法测定比格犬血浆中阿司匹林、水杨酸和单硝酸异山梨酯的浓度

目的:建立灵敏的超高效液相色谱-质谱联用法测定比格犬血浆中的阿司匹林、水杨酸和单硝酸异山梨酯的浓度。方法:选用Waters BEH C18(100 mm×2.1 mm,1.7μm)色谱柱,分别以0.5%甲酸/5 mmol.L-1醋酸铵-乙腈和2 mmol.L-1醋酸铵-乙腈为流动相,采用梯度洗脱进行分离,样品采用乙酸乙酯提取后进样,通过电喷雾电离源,以多重反应监测(MRM)方式进行负离子检测,用于定量分析的离子对分别为m/z 178.9→136.9(阿司匹林)、m/z 136.9→64.9(水杨酸)和m/z 293.7→250.0(内标,双氯酚酸钠)测定阿司匹林和水杨酸;m/z 249.6→58.9(单硝酸异山梨酯)和m/z 293.7→250.0(内标,双氯酚酸钠)测定单硝酸异山梨酯。结果:阿司匹林、水杨酸和单硝酸异山梨酯的线性范围分别为2.0～2000.0,20.0～16000.0,9.6～9557.0 ng.mL-1;定量下限分别可达2.0,20.0,9.6 ng.mL-1;日内、日间精密度(RSD)均小于15%。结论:本方法灵敏度较高,血浆用量少,适用于比格犬血浆样品中阿司匹林、水杨酸和单硝酸异山梨酯的测定和药物动力学研究。

UPLC-MS/MS determination of acetylsalicylic acid,salicylic acid and isosorbide-5-mononitrate in dog plasma

Objective:To develop an UPLC-MS/MS method for determination of acetylsalicylic acid(ASA)and its metabolite(salicylic acid,SA),isosorbide-5-mononitrate(5-ISMN) in dog plasma.Methods:The dog plasma was extracted by ethyl acetate,and separated on a BEH C18(100 mm×2.1 mm,1.7 μm)column by gradient elution using acetonitrile-0.5% formic acid-5 mmol·L-1 ammonium acetate and acetonitrile-2 mmol·L-1 ammonium acetate as mobile phases respectively.A tandem mass spectrometer equipped with electrospray ionization source was adopted as the detector.Multiple reaction monitoring(MRM)transitions of m/z 178.9→136.9,m/z 136.9→64.9,m/z 249.6→58.9 and m/z 293.7→250.0 were measured in negative mode for ASA,SA,5-ISMN and internal standard,respectively.Results:The linear concentration ranges of the calibration curve for ASA,SA,5-ISMN were 2.0-2000.0 ng·mL-1,20.0-16000.0 ng·mL-1 and 9.6-9557.0 ng·mL-1,respectively;the lower limits of quantification was 2.0,20.0,9.6 ng·mL-1,respectively;the intra-day and inter-day relative standard deviations over the entire concentration range were less than 15%.Conclusion:The method is sensitive and suitable for pharmacokinetic study of ASA,SA and 5-ISMN in dog subjects.