| 摘要: |
| 目的 研究苦参中3种生物碱成分苦参碱、氧化苦参碱、槐果碱通过Caco-2细胞的吸收特性及在大鼠肝微粒中的代谢速率。 方法 采用Caco-2细胞吸收模型分析3种生物碱的吸收特性,采用体外大鼠肝微粒体模型研究3种生物碱的代谢速率,运用HPLC-MS法测定3种生物碱类成分的含量,计算其渗透速率和代谢速率;液相条件采用Agilent Zorbax SB-C18色谱柱(3.0 mm×100 mm,3.5 μm),乙腈:水(60:40)等度洗脱,进样量5 μL,流速0.8 mL/min,柱温:30℃,运行时间9 min。质谱条件,选择正离子模式监测,干燥器温度350℃,毛细管电压4 000 V,干燥器流速 9.0 L/min,碎片电压 90 eV。 结果 Caco-2细胞吸收模型中,苦参碱、氧化苦参碱、槐果碱分别在0.050 5~10.01 μmol/L的线性范围内,线性关系良好(r> 0.999 5),通过Caco-2细胞吸收模型的渗透速率常数分别为:苦参碱1.251、氧化苦参碱0.937 5、槐果碱1.424;大鼠肝微粒体模型中,苦参碱、氧化苦参碱、槐果碱分别在0.050 1~1.001 μmol/L的线性范围内,线性关系良好(r> 0.999 8),3种生物碱类成分代谢的半衰期分别为:苦参碱4.331 h、氧化苦参碱1.084 h、槐果碱2.478 h,方法学考察结果表明,日内、日间精密度RSD%<5%,基质效应>80%,提取回收率>88%。 结论 苦参中3种生物碱类成分易通过Caco-2细胞吸收模型,主动转运为其主要的吸收方式,在大鼠肝微粒体中不易发生代谢。 |
| 关键词: 苦参 苦参碱 氧化苦参碱 槐果碱 吸收 代谢 |
| DOI：10.3724/SP.J.1008.2015.1167 |
| 投稿时间：2015-03-14修订日期：2015-07-08 |
| 基金项目:国家自然科学基金青年科学基金(81303192). |
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| Three alkaloids in Radix Sophorae flavescentis: metabolic rate in rat liver microsomes and absorption behavior in Caco-2 cells |
| ZHANG Hai1,2,ZHANG Wen1,SUN Feng-feng1,YU Jing1,CHAI Yi-feng2,ZHANG Guo-qing1* |
(1. Department of Pharmacy, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai 200438, China;
2. Department of Pharmaceutical Analysis, School of Pharmacy, Second Military Medical University, Shanghai 200433, China
*Corresponding author.) |
| Abstract: |
| Objective To investigate the absorption behaviors of three kinds of sophora alkaloids(matrine, oxymatrine and sophocarpine)in Caco-2 cells and their metabolic rates in rat liver microsomes. Methods Caco-2 cell absorption model was used to analyze the absorption behaviors of three sophora alkaloids and rat liver microsomal model in vitro was used to investigate the metabolic rates of three sophora alkaloids. The contents of three kinds of sophora alkaloids were determined by HPLC-MS and the permeation rates and metabolic rates were calculated. The analytical conditions were as follows: chromatographic column: Agilent Zorbax SB-C18(3.0 mm×100 mm, 3.5 μm); mobile phase: acetonitrile/water(60:40, V/V)with isocratic elution; injection volume: 5 μL; flow rate: 0.8 mL/min; temperature of column: 30℃; and running time: 9 min. The selective ion monitoring(SIM)in positive ion mode was used in mass spectrometry, with drying gas temperature: 350℃; capillary voltage: 4 000 V; drying gas flow rate: 9.0 L/min; and fragmentor voltage: 90 eV. Results In the Caco-2 cell absorption model, matrine, oxymatrine and sophocarpine were separated with good linearity(r> 0.999 5)between 0.050 5-10.01 μmol/L, with the permeation rates being: matrine 1.251, oxymatrine 0.937 5, and sophocarpine 1.424. In rat liver microsomal model, matrine, oxymatrine and sophocarpine were separated with good linearity(r> 0.999 8)between 0.050 1-1.001 μmol/L, with the half-life period being: matrine 4.331 h,oxymatrine 1.084 h,and sophocarpine 2.478 h. The intra-day and inter-day precisions were <5%, the matrix effect was >80%, and the extraction recovery rate was >88%. Conclusion The three kinds of sophora alkaloids can easily pass the Caco-2 cell absorption model, with active transport being the main way, and it is difficult for them to be metabolized in rat liver microsomes. |
| Key words: Sophora flavescens matrine oxymatrine sophocarpine absorption metabolism |
