基于超高效液相色谱—串联质谱同时测定大鼠血浆中杜仲制剂的药代动力学研究

Study on pharmacokinetics of Eucommia ulmoides preparation in rat plasma by UPLC-MS/MS

目的 建立用于血浆中杜仲各成分含量测定的超高效液相色谱—串联质谱的方法,并进行杜仲及其制剂各成分的药代动力学研究。方法 SD大鼠口服不同的杜仲制剂后,于不同的时间点收集血浆样品,以0.1%甲酸水溶液(A)-0.1%甲酸乙腈(B)为流动相,梯度洗脱,采用安捷伦SB C_(18)色谱柱(2.1 mm×50 mm, 1.8μm)进行分离;在多反应监测模式下,负离子模式测定;通过Phoenix Winnonlin 8.1软件计算药代动力学参数。结果 原儿茶酸、京尼平苷酸、松脂醇二葡萄糖苷、无梗五加苷、松脂醇葡萄糖苷、京尼平苷、绿原酸、咖啡酸分别在3.2-2000,3.3-2044,0.6-2028, 0.6-1770,0.3-2036,0.6-2004,0.5-1596,0.7-1090 ng/mL范围内,线性良好;专属性、准确度、精密度、稳定性、回收率和基质效应均符合生物样本检定要求。采用非房室模型分析,8个成分在AUC_(0-∞)、t_(1/2z)、T_(max)、C_(max)、V_(z)/F、Cl_(z)/F、MRT_(0-∞)方面均不相同。结论 该方法灵敏度高、专属性强、准确性好,可用于杜仲成分在大鼠体内的药代动力学研究。不同杜仲制剂在大鼠体内的释药速度、起效快慢及作用强度均不相同,在临床应用上具有指导意义。

Objective To etablish a method of ultra high performance liquid chromatography tandem mass spectrometry for the determination of the components of Eucommia ulmoides in plasma,and to study the pharmacokinetics of the components of Eucommia ulmoides and its preparation.Methods After SD rats took orally different Eucommia ulmoides preparations,plasma samples were collected at different time points,and Agilent sb C_(18) chromatographic column(2.1)was used(2.1 mm×50 mm,1.8μm).The mobile phase was consisted of 0.1%formic acid water(A)and 0.1%formic acid acetonitrile(B),and gradient elution was selected;in the multi reaction monitoring(MRM)mode,the dosage forms were measured simultaneously in the negative ion modes;the pharmacokinetic parameters were calculated by Phoenix winnonlin 8.1 software.Results Protocatechuic acid,geniposide,pinoresinol diglucoside,acanthopanax senticosus glycoside,pinoresinol glucoside,geniposide,chlorogenic acid and caffeic acid were in the range of 3.2-2000,3.3-2044,0.6-2028,0.6-1770,0.3-2036,0.6-2004,0.5-1596 and 0.7-1090 ng/mL respectively;specificity,accuracy,precision,stability,recovery and matrix effect all meet the requirements of biological sample verification.Using the non atrioventricular model analysis,the eight components were different in AUC_(0-∞)、t_(1/2z)、T_(max)、C_(max)、V_(z)/F、Cl_(z)/F、MRT_(0-∞).Conclusion The method was high sensitivity,specificity and accuracy.It can be used for the pharmacokinetics of Eucommia ulmoides in rats.Different Eucommia ulmoides preparations have different drug release speed,onset speed and action intensity in rats,which has guiding significance in clinical application.