梓葛冻干粉针中葛根素在正常大鼠和脑缺血大鼠的药代动力学比较研究  

Pharmacokinetic Study of Puerarin of Zige Freeze-Dried Powder in Cerebral Ischemia Rats and Normal Rats

作　　者：王琴[1,2,3] 邢茂[4] 唐勤[2,3] 唐清[2,3] 徐晓玉[2,3] 

机构地区：[1]重庆市中医院药物研究室,重庆市中医特色诊疗工程技术研究中心,重庆400020 [2]西南大学中医药学院 [3]西南大学中药研究所,重庆市药效评价工程技术研究中心,重庆400715 [4]第三军医大学新桥医院药剂科,重庆400037

出　　处：《中国药学杂志》2014年第7期583-587,共5页Chinese Pharmaceutical Journal

基　　金：重庆市科技攻关项目(CSTC,2010AC5011);重庆市卫生局中医药科技项目(No.2010-2-148);重庆市卫生局中医药科技项目(No.2011-2-55)

摘　　要：目的对比研究梓葛冻干粉针中葛根素在正常和脑缺血大鼠体内的药动学。方法建立测定大鼠血浆中葛根素的高效液相色谱．质谱联用法，以染料木素为内标，采用DiamonsilC18（4．6mm×150mm，5μm）色谱柱，以甲醇（含0．1％甲酸）-水（含10mmol·L-1醋酸铵）（80：20）为流动相，流速0．6mL·min-1；采用电喷雾离子源负离子模式（ESI-），多反应监测（MRM）方式对葛根素（m／z415．1→295．1）和内标物（m／z269．2→133．1）进行测定。线栓法制备永久性脑缺血模型大鼠，正常和模型大鼠分别尾静脉注射给予梓葛冻干粉针（26．7mg·kg-1），HPLC—MS／MS测定给药后不同时间点（3，5，10，30，60，120，180，300min）血浆中葛根素浓度，数据经DAS2．1软件计算药动学参数并用SPSS软件对数据进行统计学处理。结果血浆中葛根素浓度在0．02～100μg·mL-1内，呈良好线性（r=0．9954）；方法提取回收率大于86．3％，批内与批间精密度（RSD）均小于13．2％，准确度（RE）在-7．3％～10．5％内。正常组血浆中葛根素的实测达峰浓度（ρmax）为（56．5±3．4）μg·mL-1，经-时曲线下面积（AUC0-t）为（17．1±1．5）μg.h.mL-1，半衰期（t1／2）为（0．311±0．133）h，平均滞留时间（MRT0-t）为（0．269±0．044）h；在模型组，ρmax为（62．3±14．0）μg·mL-1，AUC0-t为（20．0±6．8）ng·h·mL-1，t1/2为（0．755±0．128）h，MRT0-t为（0．360±0．045）h。与正常组相比，梓葛冻干粉针中葛根素在模型组血浆中的t1/2、MRT0-t均显著延长（P〈0．05）。结论建立的HPLC—MS／MS方法专属性强、灵敏度高、快速准确，可用于大鼠血浆中葛根素的测定和药动学研究。与正常大鼠相比，梓葛冻干粉针中葛根素在永久性脑缺血模型大鼠的生物利用度提高．消除减慢、滞留时间延长。OBJECTIVE To investigate the pharmacokinetics of puerarin of Zige freeze-dried powder in the plasma of the normal rats and the model rats with middle cerebral artery occlusion（MACO）. METHODS A high-performance liquid chromatography-tan- dem mass spectrometry （HPLC-MS/MS）method was developed and validated for quantification of puerarin in rat plasma after injection administration of Zige freeze-dried powder. The plasma samples were pretreated with methanol for protein precipitation and the superna- tants were analysis on a Diamonsil C18 column （4. 6 mm × 150 mm, 5 μm）with the mobile phase consisting of 0. 1% formic acid in methanol and 10 mmol . L -1ammonium acetate in water （80： 20） at a flow rate of 0. 6 mL .min-1 to separate puerarin and genistein （internal standard, IS）. A triple-quadrupole tandem mass spectrometer under a multiple reaction-monitoring （MRM）mode to deter- mine puerarin（ m/z 415. 1→295. 1 ） and genistein（m/z 269. 2→133. 1 ）. The Zige freeze-dried powder was administered to rats in 26. 7 mg . kg-1 doses via intravenous （iv） injection, the plasma sample was collected at each time point （3, 5, 10, 30 min, 1,2, 3 and 5 h post-administration）. The pharmacokinetic analyses of puerarin in rat plasma were performed via the proprietary DAS 2. 1 computer software package. RESULTS The calibration curves had good linear in 0. 02 - 100μg . mL-1 with r = 0. 995 4. The intra- and inter-day precisions were found to be less than 13.2% , the accuracy was ranged in -7.3% - 10. 5% , and the extraction recoveries of the puerarin from the plasma was 86. 3%. The contents of puerarin arrived the max values with （56. 5 ±3.4） and （62.3 ± 14. 0）μg . mL-1, the t1/2 were （0.311±0.133） and （0.755±0.128） h, the values of AUC0-t were （17.1±1.5） and （20.0±6.8）μg. h . mL -1 , the values of MRT0-t, were （ 0. 269 ±0. 044 ） and （ 0. 360 ± 0. 045 ） h in the plasma of the normal and the model respectively. Compared with the normal, the t1/2 ,MR

关 键 词：梓葛冻干粉针 葛根素 高效液相色谱-质谱联用 血浆 药动学 

分 类 号：R285[医药卫生—中药学]