中国中药杂志

2013, v.38(05) 753-756

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RP-HPLC测定theacrine的血药浓度
Determination of theacrine in rat plasma by RP-HPLC

张维库;续洁琨;胡界晴;王夙博;李平;栗原博;姚新生;唐炳华;
ZHANG Wei-ku1,XU Jie-kun2*,HU Jie-qing2,WANG Su-bo3,LI Ping1,HIROSHI Kurihara4, YAO Xin-sheng4,TANG Bing-hua2*(1.Department of Pharmacology,Institute of Clinical Medical Sciences,China-Japan Friendship Hospital,Beijing 100029,China; 2.School of Preclinical Medicine,Beijing University of Chinese Medicine,Beijing 100029,China; 3.National Center for Pharmaceutical Screening,Institute of Materia Medica,Chinese Academy of Medical Sciences & Peking Union Medical College,Beijing 100050,China; 4.Institu

摘要(Abstract):

目的:测定theacrine灌胃后在大鼠血浆中的血药浓度。方法:采用RP-HPLC检测血浆样品,以咖啡因为内标。色谱柱为Phenomenex Luna C18(4.6 mm×250 mm,5μm),甲醇-水(25∶75)为流动相,流速为1.0 mL.min-1,检测波长为290nm,柱温为25℃。结果:Theacrine血浆中质量浓度在0.5~100 mg.L-1线性关系良好(R2=0.998 9),最低定量限为0.5 mg.L-1。在theacrine血浆质量浓度为0.5,5.0,50 mg.L-1时,日内、日间精密度均小于15%,提取回收率分别为(95.8±8.6)%,(92.5±5.0)%,(90.3±5.2)%。当给药质量浓度为30 mg.kg-1时,主要药动学参数为Cmax(35.45±2.68)mg.L-1,tmax(0.51±0.13)h,t1/2(3.13±1.37)h,AUC0-∞(265.39±94.71)mg.L-1.h。结论:经过系统的方法学考察,该研究所建立的测定方法灵敏度较高、重复性良好,可为theacrine在体内的代谢研究提供依据,为探讨theacrine作用机制及药效学评价奠定基础。
Objective: To establish a method for the determination of theacrine in rat plasma after ig.administration of theacrine.Method: Blood sample was taken timely from the eyes canthus of rats.Plasma was isolated and the protein was precipitated by ethyl acetate.Then the plasma concentration of theacrine was determined with RP-HPLC.Caffeine was used as the internal standard.The chromatographic conditions were as follows: Phenomenex Luna C18(4.6 mm×250 mm,5 μm) at 25 ℃,a mixture of methanol-water(25∶75) as the mobile phase,at the flow rate of 1.0 mL·min-1 and the detection wavelength of 290 nm.Result: The linear range of theacrine was 0.5-100 mg·L-1(R2=0.998 9).The lower limit of quantification was 0.5 mg·L-1.The intra-day RSD was 1.49%-4.40% and inter-day RSD was 0.80%-10.27%.The average extraction recoveries of theacrine were 90.3%-95.8% at concentrations of 0.5,5.0,50 mg·L-1.The main pharmacokinetic parameters after ig.administration of theacrine at concentration of 30 mg·kg-1were as follow: Cmax(35.45±2.68) mg·L-1,tmax(0.51±0.13) h,t1/2(3.13 ± 1.37) h,AUC0-∞(265.39±94.71) mg·L-1·h.Conclusion: The method has been confirmed to be simple,stable,reproducible and with high specificity,and can be used for the pharmacokinetic study of theacrine in rats.

关键词(KeyWords): theacrine;高效液相色谱法;血药浓度
theacrine;RP-HPLC;plasma concentration

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81274115,81102343,30901998);; 北京市科技新星项目(2009A35);; 教育部新世纪优秀人才项目(NCET-11-0606);; 北京中医药大学自主选题中青年教师项目

作者(Author): 张维库;续洁琨;胡界晴;王夙博;李平;栗原博;姚新生;唐炳华;
ZHANG Wei-ku1,XU Jie-kun2*,HU Jie-qing2,WANG Su-bo3,LI Ping1,HIROSHI Kurihara4, YAO Xin-sheng4,TANG Bing-hua2*(1.Department of Pharmacology,Institute of Clinical Medical Sciences,China-Japan Friendship Hospital,Beijing 100029,China; 2.School of Preclinical Medicine,Beijing University of Chinese Medicine,Beijing 100029,China; 3.National Center for Pharmaceutical Screening,Institute of Materia Medica,Chinese Academy of Medical Sciences & Peking Union Medical College,Beijing 100050,China; 4.Institu

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