HPLC-MS/MS法定量研究比格犬口服替米沙坦后的血浆药代动力学

Pharmacokinetic Study of Telmisartan in Beagle Dogs Using HPLC-MS /MS Analysis

目的建立比格犬血浆中替米沙坦的HPLC-MS/MS定量测定方法。方法液相色谱分离采用Zorbax SB-C18色谱柱,流动相为乙腈-4 mmol·L-1醋酸铵水（40∶60,0.1%甲酸）,流速为0.2 ml·min-1;质谱检测采用电喷雾（ESI）离子源,正离子多反应监测（MRM）方式检测。血浆样品经乙腈沉淀蛋白、高速离心后进样分析,检测反应为m/z 515.2-276.3（替米沙坦）,m/z 256.2-167.0（苯海拉明,内标）。结果替米沙坦在比格犬血浆中定量范围为0.5~2000 ng·ml-1,日内和日间精密度≤9.13%、回收率满足血浆测定要求,且血浆样品稳定性好。结论本方法特异、快速、准确、可靠,可满足替米沙坦比格犬血浆动力学研究需要。

Objective To develop an HPLC-MS/MS method for the quantification of telmisartan in plasma of Beagle dogs. Methods Telmisartan and diphenhydamine（internal standard,IS） were separated on a Zorbax SB-C 18 column with a mobile phase of acetonitrile-water（40∶ 60） containing 0. 1% formic acid and 4 mmol·L1ammonia acetate at a flow rate of 0. 2 ml·min-1. The plasma sample was prepared by a simple deproteinization with acetonitrile followed by centrifugation. The detection was performed in a positive multiple reaction monitoring（MRM） mode with an electrospray ionization（ESI） source. The reactions monitored were m/z 515. 2-276. 3 for telmisartan and m/z 256. 2-167. 0 for diphenhydamine（IS）. Results The quantification range for telmisartan in Beagle dog plasma was 0. 5-2000 ng·ml-1. The intra-and inter-day precision（RSD%） was no more than 9. 13% with high recovery and good stability for quantification. Conclusion This method is specific,quick,accurate and reliable for the pharmacokinetic study of telmisartan in Beagle dogs.