基于UPLC-MS/MS的蒽贝素血药浓度测定及其在大鼠体内药动学研究

Determination of Embelin Blood Concentration and Its Pharmacokinetic Study in Rats by UPLC-MS/MS

关键词:  蒽贝素  大鼠  血药浓度  超高效液相色谱串联质谱法  药动学

Key words:  embelin  rat  plasma concentration  UPLC-MS/MS  pharmacokinetics

摘要:

建立测定大鼠血浆中蒽贝素（Embelin）浓度的方法，并进行药动学研究。选取12只Sprague-Dawley（SD）大鼠灌胃蒽贝素（15.0 mg/kg），分别于给药前及给药后0.083，0.25，0.50，0.75，1，2，3，4，6，8，10，12，24，36，48，72 h时眼底静脉采血，分离血浆，用丙酮-乙酸乙酯混合溶剂萃取后，以大黄素为内标，采用超高效液相色谱串联质谱法（UPLC-MS/MS）测定。色谱柱为Agilent Zorbax Eclipse Plus C18，流动相为甲醇-0.1%氨水溶液，梯度洗脱，流速为0.3 mL/min。样品经电喷雾离子源电离为负离子化后，在多反应监测模式下测定蒽贝素（m/z 293.1→96.6）和内标物大黄素（m/z 269.0→225.1）的浓度，并计算药动学参数。研究结果显示：蒽贝素检测血药浓度的线性范围为10—1 200 ng/L，定量下限为10 ng/mL，检出限为3 ng/mL，日内和日间RSD均小于10%，准确度为8.8%—14.0%。大鼠灌胃蒽贝素的平均药-时曲线符合二室模型，分布半衰期为（12.60±1.19）h，消除半衰期为（15.95±0.73）h，AUC_0-t为（3 596.31±271.93）ng·h/L，AUC_0-∞为（3 717.48±269.82）ng·h/L。蒽贝素在大鼠体内的药动学过程符合二室模型。本研究建立的方法简单、快速、准确、选择性强，可用于蒽贝素血药浓度的测定及药动力学研究。

Abstract:

To establish a method for concentration determination of embelin in rat plasma,and to conduct its pharmacokinetics study.Twelve Sprague-Dawley (SD) rats were gastric infusion with embelin solution (15.0 mg/kg) via fundus vein. Blood sample were collected before medication and 0.083，0.25，0.50，0.75，1，2，3，4，6，8，10，12，24，36，48，72 h after medication. After the plasma isolated and extracted with a mixed solution of acetone and ethyl acetate，UPLC-MS/MS method was adopted using emodin as internal standard. The chromatographic column was Agilent Zorbax Eclipse Plus C18,the mobile phase was methanol-0.1% ammonia solution,gradient elution,and the flow rate was 0.3 mL/min. Embelin and emodin were measured by ESI in negative electron mode using multiple reaction monitoring (MRM),and the pharmacokinetic parameters of embelin were calculated. The extracted ions monitored following MRM transitions were m/z 293.1→96.6 for embline and m/z 269.0→225.1 for internal standard emodin. The linear range of embline detection blood concentration was 10-1 200 ng/L. The lower limit of quantification was 10 ng/mL,and the detection limit was 3 ng/mL. RSDs of intra-day and inter-day were both less than 10%.The accuracy ranged from 8.8% to 14.0%.The average plasma-time curve of embline with gastric infusion in rats was in line with the two-compartment model. The distribution half-life of all rats was (12.60±1.19) h，and the elimination half-life was (15.95±0.73) h. AUC_0-t was (3 596.31±271.93) ng·h/L,and AUC_0-∞ was (3 717.48±269.82) ng·h/L.Pharmacokinetic study showed that the pharmacokinetic process of the compound is in line with two-compartment model in rats.The method established in this study is simple,rapid,accurate and highly selective,and can be used for plasma content determination of embelin and pharmacokinetic studies.