基于自动采血系统与HPLC-QQQ-MS的青藤碱药动学分析  

Pharmacokinetic Analysis of Sinomenine Based on Automatic Blood Sampling System and HPLC-QQQ-MS

作　　者：赵小亮[1] 李涛[1] 刘洋[1] 张美玉[1] 陈跃[2] 崔月[1] 张莹[1] 孙丹丹[1] 王志国[1] 王丹巧[1] 

机构地区：[1]中国中医科学院医学实验中心,北京市中医药防治重大疾病基础研究重点实验室,北京100700 [2]山东省分析测试中心,济南250014

出　　处：《中国实验方剂学杂志》2015年第14期66-70,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家科技部国际科技合作项目(2010DFA31890);中国中医科学院自主选题(zz2012006)

摘　　要：目的：建立灵敏、简便、准确的自动采血系统采集大鼠血浆样本,高效液相色谱-三重四级杆质谱联用技术（HPLC-QQQ-MS）测定血浆中青藤碱含量的方法,并应用于其体内药代动力学研究。方法：SD大鼠颈总静脉插管,待清醒后按40 mg·kg-1腹腔注射青藤碱,应用自动采血系统在大鼠自由活动状态下采集样品。血浆样品中加入内标（磷酸可待因）,乙腈沉淀蛋白,流动相乙腈-水（含0.1%甲酸铵）梯度洗脱。质谱采用电喷雾电离源（ESI）,扫描方式为多反应离子监测模式（MRM）,定量分析离子对分别为青藤碱m/z 330.0,磷酸可待因m/z 300.0。利用Win Nonlin软件计算药代动力学参数。结果：血浆中青藤碱在5.0~500.0μg·L-1线性关系良好,定量下限1μg·L-1,样品处理方法的提取回收率〉88%,日内及日间准确度101.08%~111.08%,RSD均〈10%,稳定性良好。血浆中青藤碱的达峰时间（tmax）,药峰浓度（Cmax）,药时曲线下面积（AUC0-24 h）,表观分布容积（Vz/F）,清除率（CL/F）,平均驻留时间（MRT0-24 h）分别为（1.65±1.48）h,（210.43±96.35）μg·L-1,（1 053.80±427.69）h·μg·L-1,（252.65±57.36）L·kg-1,（30.37±11.85）L·h-1·kg-1,（5.15±2.47）h。结论：建立的方法有效控制了采血过程对药动学分析结果的影响,样品用量少,简便、准确、可靠,能够满足自由活动大鼠体内青藤碱连续定量分析和药代动力学研究。Objective： To develop a sensitive and acurate automatic blood sampling system and HPLC- QQQ-MS method for determining the content of sinomenine and its pharmacokinetic characteristics in rat plasma. Method： SD rats were jugular vein cannula and intraperitoneal injected with 40 mg·kg-1 sinomenine after sober, blood samples were collected in rats at freely moving state by automatic blood sampling system. Codeine phosphate was used as the internal standard calibration in 20 μL sample, acetonitrile for protein precipitation. Gradient elution was performed on a Zorbax XBD-C18 column, mobile phase was consisted of 0. 1% ammonium formate and acetonitrile. The analyte was detected using electrospray ionization （ESI） in multiple reaction monitoring （MRM） modes. Reaction selected ions were m/z 330.0 for sinomenine and m/z 300.0 for internal standard.Pharmacokinetic parameters were calculated by Phoenix WinNonlin software. Result： Linear ranges of sinomenine was 5.0-500.0 μg ·L-1, extract recovery of this method was more than 88% , the lower limit of quantification was 1 μg·L-1 , intra-day and inter-batch accuracy was 101.08% - 111.08% with RSD were less than 10%. Stability was well. Main pharmacokinetic parameters of t C AUC0aa h, Vz/F, CL/F and MRT0_24 hwere （ 1.65 ± 1.48） h, （210.43 ±96.35） μg·L-1, （1 053.80 ±427.69） h·μg·L-1, （252.65 ±57.36） L·kg-1, （30. 37 ±11.85） L·h-1·kg-1and （5. 15 ± 2.47） h, respectively. Conclusion： It is first time to establish a method using automatic blood sampler and HPLC-QQQ-MS to collect blood sample and determine the concentration of sinomenine in rat plasma. This method needs less sample quantity. It is so easy, acurate and sensitive for using in in vivo quantitative analysis of sinomenine.

关 键 词：青藤碱 自动采血系统 自由活动大鼠 药代动力学 高效液相色谱-三重四级杆质谱联用技术 

分 类 号：R969.1[医药卫生—药理学] R284.1[医药卫生—药学]