高效液相色谱-串联质谱法同时测定大鼠血浆中阿托伐他汀钙和非洛地平的浓度

Simultaneousdetermination of atorvastatin calcium and felodipine in rat plasma by LC-MS/MS

目的:建立快速灵敏高效液相色谱-串联质谱(HPLC-MS/MS)法同时测定大鼠血浆中阿托伐他汀钙和非洛地平的浓度,并研究其单用及联用时在其体内的药动学特征。方法:色谱柱:Diamonsil ODS C_(18)(150mm×4.6mm,5μm);流动相:甲醇-0.1%甲酸水溶液(70∶30,V/V);流速:0.8mL·min^(-1),进样量10μL。采用ESI+源,选择性离子检测方式(SIM),对离子反应m/z559.2(阿托伐他汀钙),m/z406.0(非洛地平),m/z325.2(内标为甲基睾酮)进行检测。结果:阿托伐他汀钙和非洛地平在0.01~0.8μg·mL^(-1)范围内线性关系良好,定量下限0.01μg·mL^(-1),精密度、稳定性均符合要求。与单用组相比,药物联用组阿托伐他汀钙的最大血药浓度由(677.74±100.86)μg·L^(-1)增加到(789.80±12.16)μg·L^(-1),血浆药物-时间曲线下面积由(5 755.90±1 210.84)h·μg·L^(-1)增加到(6 931.74±228.11)h·μg·L^(-1),半衰期由(3.77±0.89)h变为(3.60±0.36)h;联用组中非洛地平的最大血药浓度由(492.40±69.30)μg·L^(-1)上升到(751.50±26.12)μg·L^(-1),血浆曲线下面积由(4 150.66±725.61)h·μg·L^(-1)增加到(6 854.87±725.61)h·μg·L^(-1),半衰期由(2.64±0.20)h延长至(2.88±0.23)h。结论:该方法灵敏、准确、可靠,专属性强,适用于阿托伐他汀钙和非洛地平在大鼠体内的药动学研究。

OBJECTIVE To establish a sensitive,specific and fast LC-MS/MS method for simultaneous determination of atorvastatin calcium and felodipine in rat plasma,and study the pharmacokinetic interaction between atorvastatin calcium and felodipine in rats.METHODS Chromatographic separation was performed by pumping the moile phase into a Diamonsil ODS C_（18）（150 mm×4.6 mm,5μm）.Mobile phase was composed of methanol and water（0.1%formic acid,70∶30,V/V）at a flow rate of 0.8 mL·min^-1.Injection volume was 10μL aliquot of solution.The precursor-fragments of atorvastatin calcium,felodipine and methyl testosterone were m/z559.2,m/z406.0 and m/z325.2,respectively,and were detected with the ESI source in positive ion and the selected ion monitoring（SIM）mode.RESULTS The range of calibration curve was 0.01-0.8μg·mL^-1 with a good correlation coefficient.The lowest limit of quantification was 0.01μg·mL^-1.The intra and inter-batch precision values of the method were less than 15%.Combined treatment with atorvastatin calcium and felodipine increased the peak concentration of atorvastatin calcium from（677.74±100.86）μg·L^-1 to（789.80±12.16）μg·L^-1.The AUC（0-∞）value of atorvastatin calcium increased from（5 755.90±1 210.84）h·μg·L^-1 to（6 931.74±228.11）h·μg·L^-1.The t1/2 values were the opposite.The Cmax,AUC（0-∞）and t1/2 values of felodipine in combination group were greater than the values of the individual group.CONCLUSION The method is fast,simple and sensitive,and suitable for determination of accurate concentrations and pharmacokinetics of atorvastatin calcium and felodipine in rat plasma.