中国中药杂志

2007, (01) 57-60

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LC-MS法测定槐果碱及其在Caco-2细胞上的转运特征
Study on transport of sophocarpine in millicell system by LC-MS

王素军;王广基;李晓天;阮金秀;张振清;
WANG Su-jun~(1),WANG Guang-ji~(2),LI Xiao-tian~(2),RUAN Jin-xiu~(1),ZHANG Zhen-qing~(1)(1.Institute of Pharmacology and Toxicology,Academy of Military Medical Sciences,Beijing 100850,China;2.Key Lab of Drug Metabolism and Pharmacokinetics,China Pharmaceutical University,Nanjing 210038,China)

摘要(Abstract):

目的:建立槐果碱的LC-MS测定法,研究其在Caco-2细胞上的转运特征。方法:采用Zorbax Extend-C18柱,流动相10 mmol.L-1醋酸铵水溶液-乙腈(85∶15),流速0.2 mL.m in-1。电喷雾离子化(ESI)方式,采用选择性离子检测法,检测离子为正离子,槐果碱的选择性离子为[m/z+H]+247.3;考察了温度、药物浓度、时间对槐果碱在Caco-2细胞上转运的影响。结果:槐果碱在52 000μg.L-1呈良好的线性关系(r=0.998 5),最低定量限为5μg.L-1;槐果碱在M illicell系统上的转运量具有一定的浓度-时间依赖性,其转运表现为被动扩散,槐果碱的平均表观通透系数Papp为3.159×10-5cm.s-1。结论:该法灵敏、简单,专属性强,槐果碱在Caco-2细胞上具有良
Objective: To establish LC-MS method for determination of sophocarpine(SC) and investigate its transport in millicell system.Method: Separation of SC was carried out by an extend-C_(18) column(2.1 mm×50 mm,5 μm,agilent technologies,USA).The mobile phase was acetonitrile-water containing 10 mmol·L2 000μg.L-1呈良好的线性关系(r=0.998 5),最低定量限为5μg.L-1;槐果碱在M illicell系统上的转运量具有一定的浓度-时间依赖性,其转运表现为被动扩散,槐果碱的平均表观通透系数Papp为3.159×10-5cm.s-1。结论:该法灵敏、简单,专属性强,槐果碱在Caco-2细胞上具有良
Objective: To establish LC-MS method for determination of sophocarpine(SC) and investigate its transport in millicell system.Method: Separation of SC was carried out by an extend-C_(18) column(2.1 mm×50 mm,5 μm,agilent technologies,USA).The mobile phase was acetonitrile-water containing 10 mmol·L(-1) ammonium acetate(15∶85) at the flow rate of 0.2 mL·min(-1) ammonium acetate(15∶85) at the flow rate of 0.2 mL·min(-1).The ESI ion source was set in the positive ion polarity mode for acquiring all mass spectrometry data.The selective ion monitoring(SIM) was used,and protonated molecules (-1).The ESI ion source was set in the positive ion polarity mode for acquiring all mass spectrometry data.The selective ion monitoring(SIM) was used,and protonated molecules (+) were detected at m/z 247.3 for SC.This LC-MS method was applied to determine SC concentration in millicell system.Result: The linearity of SC ranged from 5 to 2 000 μg·L(+) were detected at m/z 247.3 for SC.This LC-MS method was applied to determine SC concentration in millicell system.Result: The linearity of SC ranged from 5 to 2 000 μg·L(-1)(r=0.998 5).The quantification limit of SC was 5 μg·L(-1)(r=0.998 5).The quantification limit of SC was 5 μg·L(-1).SC transport in Caco-2 cells was time and concentration dependent,and SC transport complied with the passive diffusion.Its average apparent permeability coefficient(P_(app)) is 3.159×10(-1).SC transport in Caco-2 cells was time and concentration dependent,and SC transport complied with the passive diffusion.Its average apparent permeability coefficient(P_(app)) is 3.159×10(-5) cm·s(-5) cm·s(-1).Conclusion: This assay was highly sensitive and specific enough for determining concentrations of SC in millicell system,and SC showed good transport in millicell system.

关键词(KeyWords): 槐果碱;LC-MS;Caco-2细胞;转运
sophocarpine;liquid chromatography-mass spectrometry;Caco-2 cells;transport

Abstract:

Keywords:

基金项目(Foundation): 国家863计划(2003AA2Z347A)

作者(Authors): 王素军;王广基;李晓天;阮金秀;张振清;
WANG Su-jun~(1),WANG Guang-ji~(2),LI Xiao-tian~(2),RUAN Jin-xiu~(1),ZHANG Zhen-qing~(1)(1.Institute of Pharmacology and Toxicology,Academy of Military Medical Sciences,Beijing 100850,China;2.Key Lab of Drug Metabolism and Pharmacokinetics,China Pharmaceutical University,Nanjing 210038,China)

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