高效液相色谱-荧光法测定大鼠血浆中富马酸比索洛尔的浓度及药物动力学

Determination of bisoprolol fumarate and its pharmacokinetics in rat plasma by HPLC-fluorescent method

目的建立大鼠血浆中富马酸比索洛尔（BF）浓度的测定方法,并研究大鼠灌胃给予BF后的体内药动学。方法以酒石酸美托洛尔为内标,采用HPLC-荧光法,测定大鼠血浆中药物浓度。色谱柱：Wondasil C18（250mm×4.6 mm,5μm）;流动相：乙腈-0.01 mol·L-1磷酸氢二铵缓冲液（35：65）;流速：1.0 mL·min-1;荧光检测的激发波长与发射波长分别为275 nm与305 nm;柱温：室温。大鼠灌胃给予0.45 mg·kg-1的BF后,于不同时间采血,测定血浆中药物浓度并估算其药动学参数。结果 BF在4.0--128.0 ng·mL-1与峰面积线性关系良好,r=0.999 6,提取回收率为88.52%--96.3%,定量下限为4.0 ng·mL-1。大鼠灌胃给予0.45 mg·kg-1的BF后,其体内消除半衰期为2.5 h。结论本方法操作简便、经济,准确度与灵敏度高,可用于大鼠血浆中BF的测定及药动学研究。

Objective To establish an HPLC method to determine bisoprolol fumarate（BF） in the plasma and its pharmacokinetics after oral administration in rats. Methods The concentration of BF in the rat plasma was determined by HPLC with fluorescence detector and metoprolol tartaric acid was used as the internal standard. The chromatographic conditions were as follows： Wondasil C18 column（ 250 mm×4.6 mm, 5 μm） was used and the mobile phase consisted of acetonitrile and 0.01 mol · L-1 diammonium hydrogen phosphate（35 ：65） at 1.0 mL · min-1. The excitation wavelength and emission wavelength were 275 nm and 305 nm. The column temperature was room temperature. Rat plasma samples were collected at different intervals after the administration of a single oral dose of BF（0.45 mg · kg-1） and its plasma concentration was determined by HPLC method to estimate the pharmacokinetic parameters of the drug in the rats. Results There was a good linearity for BF at 4.0-128.0 ng · mL-1（r = 0.999 6）. The extraction recovery was from 88.52% to 96.32% and the lower limit of quantitation of the method was 4.0 ng · mL-1. The elimination half life was 2.5 h after the oral administration of BF. Conclusion The method is simple, economic, accurate and sensitive for the determination of BF in the rat plasma.