HPLC-MS/MS测定大鼠血浆中普萘洛尔的含量及其药动学研究

Determination of propranolol in rat plasma by HPLC-MS/MS and its application in pharmacokinetic study

关键词:  普萘洛尔  高效液相色谱-质谱联用  大鼠  血浆  药动学

Key words:  propranolol  HPLC-MS/MS  rat  plasma  pharmacokinetics

摘要:

目的 建立大鼠血浆中普萘洛尔的高效液相色谱-质谱联用（HPLC-MS/MS）定量分析方法，考察大鼠口服给药后，普萘洛尔在其体内的药动学特征。方法 血浆样品用甲醇沉淀蛋白法处理后，以盐酸美西律为内标，采用LC-MS/MS分析方法，测定大鼠血浆中普萘洛尔的浓度。色谱柱为ZORBAX Eclipse Plus C₁₈柱（2.1 mm×100 mm，3.5 μm），流动相为0.1%甲酸水溶液-乙腈（70：30）等度洗脱；采用ESI离子源，正离子模式，多反应离子监测（MRM）m/z 260.2→116.2（普萘洛尔）和m/z 180.2→58.2（内标盐酸美西律）。结果 普萘洛尔在5~1 000 ng·mL^-1内线性关系良好；日内、日间精密度RSD为0.36%~6.39%，准确度为96.19%~113.38%；提取回收率为86.82%~96.12%；基质效应为100.1%~103.1%。结论 本方法经方法学验证，可用于大鼠血浆中普萘洛尔的测定及其药动学研究。

Abstract:

OBJECTIVE To establish an HPLC-MS/MS method to determine the content of propranolol in rat plasma, and to study the pharmacokinetic characteristics of propranolol in rats after oral administration. METHODS The plasma samples were treated with methanol precipitation protein method. LC-MS/MS method was developed and validated for the determination of propranolol in rats using mexiletin as the internal standard. ZORBAX Eclipse Plus C₁₈column (2.1 mm×100 mm, 3.5 μm) was used, the mobile phase consisting of water containing 0.1% formic acid and acetonitrile (70:30). The detection parameters of mass spectrometry were as follows:ESI ionization source, positive polarity. Ions monitored in the multiple reaction monitoring (MRM) mode were m/z 260.2→116.2 for propranolol and m/z 180.2→58.2 for mexiletin (internal standard). RESULTS Calibration curves were linear in range of 5-1 000 ng∙mL^-1 for propranolol. Intra-and inter-day precision and accuracy were 0.36%-6.39% and 96.19%-113.38%, respectively. Extraction recovery ranged 86.82%-96.12%, and the matrix effect of propranolol ranged 100.1%-103.1%. CONCLUSION The LC-MS/MS method is accurate and sensitive, and is successfully applied to evaluate the pharmacokinetics of propranolol in rats plasma.