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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(3.0mm×100mm,3.5μm),乙腈∶水(60∶40)等度洗脱,进样量5μL,流速0.8mL/min,柱温:30℃,运行时间9min。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 000V,干燥器流速9.0L/min,碎片电压90eV。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 5~10.01μmol/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 1~1.001μmol/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4.331h、氧化苦参碱1.084h、槐果碱2.478h,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。
Objective To study the absorption characteristics of matrine, matrine, oxymatrine and sophocarpine in Caco-2 cells and their metabolic rate in rat liver particles. Methods Caco-2 cell absorption model was used to analyze the absorption characteristics of three alkaloids. The in vitro rat liver microsome model was used to study the metabolic rate of three alkaloids. The contents of three alkaloids were determined by HPLC-MS. Its permeation rate and metabolic rate. The liquid phase was eluted with an Agilent Zorbax SB-C18 column (3.0 mm × 100 mm, 3.5 μm) and isocratic with acetonitrile: water (60:40). The injection volume was 5 μL and the flow rate was 0.8 mL / min, column temperature: 30 ℃, running time 9min. Mass spectrometry conditions, the choice of positive ion mode monitoring, dryer temperature 350 ℃, capillary voltage 4 000V, dryer flow 9.0L / min, debris voltage 90eV. Results In the model of Caco-2 cell uptake, matrine, oxymatrine and sophocarpine were linear in the range of 0.050 5-10.01 μmol / L (r> 0.999 5) The permeation rate constants of absorption model were as follows: matrine 1.251, oxymatrine 0.937 5 and sophocarpine 1.424, respectively. Matrine, oxymatrine and sophocarpine in rat liver microsomes were respectively 0.050 1 ~ 1.001μmol / L, the linear relationship was good (r> 0.999 8). The half-lives of three alkaloids were: matrine 4.331h, oxymatrine 1.084h, sophocarpine 2.478h The results of methodological study showed that RSD% <5%, matrix effect> 80%, extraction recovery rate> 88% during the day and the day. Conclusion The components of the three alkaloids in Sophora flavescens easily pass through the Caco-2 cell absorption model and are actively transported to the main absorption mode, which is not easy to be metabolized in the rat liver microsome.