LC-MS/MS 法测定比格犬血浆中丁酸氯维地平浓度

摘要

本研究建立了液相色谱-串联质谱(LC-MS/MS)法测定比格犬血浆中丁酸氯维地平浓度,用来研究自制的丁酸氯地平脂肪乳注射液与原研产品在比格犬体内的药代动力学参数,同时比较其生物等效性。实验以氨氯地平为内标,选取 Phenomenex Luna C8色谱柱(2.0 mm×150 mm×5μm),以甲醇-0.1%甲酸溶液(82∶18,V/V )为流动相,采用 Waters Quattro Micro API 的正离子检测方式,用MassLynx4.1软件进行数据处理。结果表明,丁酸氯维地平标准曲线的线性范围为0.4~100μg/L。主要的药代动力学参数：参比制剂和试验制剂的 C max 平均值分别为(58.748±16.738)μg/L 和(53.706±18.963)μg/L；T max 分别为(2.938±0.678)μg/L 和(2.875±0.991)μg/L；T 1/2分别为(11.88±3.824) min 和(11.587±3.634)min；AUC0→t 分别为(1883.821±647.882)μg · min/L 和(1856.541±590.653)μg· min/L；AUC0→∞分别为(1889.834±649.135)μg · min/L 和(1863.485±592.039)μg·min/L。方差分析表明,这两种制剂的主要药动学参数之间无明显差异；双单侧 t 检验结果表明,两制剂在比格犬体内为生物等效制剂。%Pharmacokinetic parameters of the cleviprex fat emulsion injection and origi-nal research products in beagle dogs plasma were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS).Taking amlodipine as internal standard, metabolites were separated on Phenomenex Luna C8(2.0 mm×150 mm×5 μm)column by the gradient elution with methanol-0.1% formic acid (82 ∶ 18,V/V )as mobile phase,and detected by positive ions electrospray ionization (ESI)mass spectrometry with fullscan-data dependent scan.Data was disposed through MassLynx4.1.The result shows that the linear range of cleviprex is 0.4-100 μg/L.The average values of C max ,T max ,T 1/2 ,AUC0→t and AUC0→∞ are (58.748 ± 1 6.738)μg/L,(2.938 ±0.678)μg/L,(1 1.88 ± 3.824 ) min,(1 883.821 ± 647.882 )μg · min/L,(1 889.834 ± 649.135)μg·min/L for the reference preparation,respectively.While the average val-ues of C max ,T max ,T 1/2 ,AUC0→t and AUC0→∞ are (53.706 ± 18.963)μg/L,(2.875 ± 0.991 )μg/L,(1 1.587±3.634)min,(1 856.541±590.653)μg·min/L,(1 863.485± 592.039)μg·min/L,for the test preparation,respectively.ANOVA and two one-sided t-tests analysis show that there is no significant difference with the pharmacokinetic parameters of the two formulations,and they have bioequivalent in Beagle dogs plasma.