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Diamonsil C18(4.6 mm×150 mm,5 μm)色谱柱,以甲醇(含0.1%甲酸)-水(含10 mmol·L-1醋酸铵)(80∶20)为流动相,流速0 6 mL·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/z 415.1→295.1)和内标物(m/z 269.2→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(26.7 mg·kg-1),HPLC-MS/MS测定给药后不同时间点(3,5,10,30,60,120,180,300 min)血浆中葛根素浓度,数据经DAS 2.1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0.02~100 μg·mL-1内,呈良好线性(r=0.995 4);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(t 1/2)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,t 1/2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t 1/2、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。
Abstract
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-tandem 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 supernatants 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-1 ammonium 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 determine 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 0.02-100 μg·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 t 1/2 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 t 1/2、MRT0-t of the model were significantly longer (P＜0.05). CONCLUSION The developed and validated method is specifically, rapid and sensitive, and has been successfully applied to research the pharmacokinetics of puerarin in rats plasma after iv zige freeze-dried powder. The pharmacokinetic parameters showed that the puerarin in the Zige freeze-dried powder has better biological availability, slower elimination and longer resistance time in the model rats, which indicate the reduce of the rate of metabolism in the model and is beneficial to the clinical pharmacokinetic studies after iv administration of Zige freeze-dried powder.
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
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Key words
Zige freeze-dried powder /
puerarin /
HPLC-MS/MS /
plasma /
pharmacokinetics
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王琴,,邢茂,唐勤,唐清,徐晓玉*.
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[J]. 中国药学杂志, 2014, 49(7): 583-587 https://doi.org/10.11669/cpj.2014.07.018
WANG Qin, XING Mao, TANG Qin, TANG Qing, XU Xiao-yu*.
Pharmacokinetic Study of Puerarin of Zige Freeze-Dried Powder in Cerebral Ischemia Rats and Normal Rats[J]. Chinese Pharmaceutical Journal, 2014, 49(7): 583-587 https://doi.org/10.11669/cpj.2014.07.018
中图分类号：
R285
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参考文献
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脚注
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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)
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