HPLC-MS法测定大鼠注射丹红注射液后血浆中3种酚酸类成分

Determnation of three kinds of phenolic acids in rat plasma after iv administration of Danhong Injection by HPLC-MS

目的建立定量测定大鼠注射丹红注射液后血浆中迷迭香酸、紫草酸、丹酚酸B的方法。方法采用HPLC-MS法,Diamonsi(l钻石)C1(8250 mm×4.6 mm,5μm)色谱柱,乙腈-0.4%甲酸水溶液为流动相,梯度洗脱;体积流量1.0 mL/min,MS检测器选用正离子模式。结果迷迭香酸在0.45～100.0μg/mL(r2=0.996 5)线性关系良好,回收率在85.0%～101.4%,日内、日间精密度的RSD<8%;紫草酸在0.45～100.0μg/mL(r2=0.995 9)线性关系良好,回收率在84.4%～93.94%,日内、日间精密度的RSD<9%;丹酚酸B在0.7～200.0μg/mL(r2=0.998 9)线性关系良好,回收率在80.0%～97.4%,日内、日间精密度的RSD<5%。结论所建立的方法灵敏度好、准确率高,可用于丹红注射液中酚酸类成分药动学研究。

Objective To establish an HPLC-MS method for determining the level of three kinds of phenolic acids （rosmarinic acid, lithospermic acid, and salvianolic acid B） in rat plasma, Methods Diamonsil C18 column （250 mm ×4.6 mm, 5 μm） was used. Acetonitdle-0.4% formic acid solution was used as the mobile phase at a flow rate of 1 mL/min, and electrospray ionization mass spectrometry （ESI-MS） was used to detect and quantify. Results Rosmarinie acid had a good linear relationship during 0.45--100.0 μg/mL （r^2 = 0.996 5）, with absolute recoveries 85.0%--101.4%. The RSD values of intra- and inter-day were less than 8%. Lithospermic acid had a good linear relationship during 0.45--100.0 μg/mL （r^2 = 0.995 9）, with absolute recoveries 84.4%--93.94%. The RSD values ofintra- and inter-day were less than 9%. Salvianolic acid B had a good linear relationship during 0.7--200.0 μg/mL （r^2 = 0.998 9）, with absolute recoveries 80.0%--97.4%. The RSD values ofintra- and inter-day were less than 5%. Conclusion The method is simple, quick, and convenient for the pharmacokinetic study on phenolic acids in Danhong Injection.