HPLC-MS法测定大鼠体内大黄素血药浓度及药动学研究

Determination of emodin in plasma by HPLC-MS and its pharmacokinetic study in rats

目的建立高效液相色谱/四极杆-静电场轨道阱高分辨质谱(HPLC-Q-HR/MS)方法,研究大黄素在正常与脑缺血大鼠体内的药动学规律。方法采用HPLC-Q-HR/MS内标法测定大黄素血药浓度,色谱柱为XBridge^(TM) C_(18)柱(150 mm×2.1mm,5μm),柱温30℃;流动相为3 mmol/L乙酸铵-甲醇,梯度洗脱,梯度洗脱程序为0～2 min,30%甲醇;2～10 min,30%～60%甲醇;10～13 min,60%～30%甲醇;体积流量为0.3 m L/min;进样体积为5μL;质谱条件:离子源为加热电喷雾离子化源(HESI),扫描方式为全扫模式,负离子模式检测。以DAS 3.0软件拟合计算药动学参数。结果大黄素在正常与脑缺血大鼠血浆中的主要药动学参数分别为AUC_(0-∞)(605.63±163.66)、(1 107.78±191.11)ng?h/m L,C_(max)(81.96±20.72)、(91.65±16.82)ng/m L,V_Z/F(851.03±97.30)、(1 051.87±119.88)L/kg,t_(1/2)(10.31±1.61)、(23.13±3.56)h,t_(max)(0.75±0.22)、(0.75±0.16)h。结论该法操作简便、分析速度快速、灵敏,适用于大黄素在大鼠体内的药动学研究。

Objective To establish a method for determination of emodin in rat plasma by HPLC/Q-Exactive HR/MS, and to study the pharmacokinetics of emodin in normal rats and cerebral ischemia rats. Methods The plasma concentration of emodin was determined by HPLC/Q-Exactive HRMS method with internal standard method. Emodin was eluted on a XBridgeTM C18 （150 mm × 2.1 ram, 5 μm） column with temperature at 30℃. The mobile phase consisted of 3 mmol/L ammonium acetate and methanol, with a gradient program as follows： 0-2 min （30% methanol）, 2--10 min （30%--60% methanol）, 10--13 min （60%--30% methanol）. The flow rate was 0.3 mL/min, and the injection volume was 5 pL. MS experiments were coupled with the HPLC via HESI source operated in negative ionization full-scan mode. The pharmacokinetic parameters were calculated by the sottware of DAS 3.0. Results The main pharmacokinetic parameters of emodin in normal rats and cerebral ischemia rats were as follows： AUC0.~ were （605.63± 163.66） and （1 107.78 ± 191.11） ng.h/mL, Cmax were （81.96 ± 20.72） and （91.65 ± 16.82） ng/mL, Vz/F were （851.03± 97.30） and （1 051.87± 119.88） L/kg, tl/2 were （10.31 ± 1.61） and （23.13 ± 3.56） h, tmax were （0.75±0.22） and （0.75± 0.16） h. Conclusion The method is simple, accurate, fast, sensitive, and suitable for the pharmacokinetic study of emodin in rats.