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目的研究染料木黄酮在Caco-2单细胞层模型上的吸收特征。方法将染料木黄酮的细胞样品经高速离心后取上清液,以乙腈-醋酸铵水溶液为流动相,以睾丸酮为内标,采用超高效液相色谱(ultra performance liquid chromatography,UPLC)法检测。考察时间、pH值等因素对染料木黄酮吸收的影响,测定透过Caco-2单细胞层的染料木黄酮浓度并计算其表观渗透系数(Papp)。结果 Caco-2细胞对染料木黄酮的吸收在1 h内呈线性,药物摄取时间定为1 h;pH值对染料木黄酮的吸收没有显著影响。染料木黄酮的线性范围为0.625~40μmol/L,日间、日内精密度均小于15%。Papp从基顶侧(apical,AP)到基底侧(basolateral,BL)为1.51×10 5cm/s,从BL侧到AP侧为1.84×10 5cm/s,其渗透系数Papp(BL-AP)/Papp(AP-BL)为1.22。结论染料木黄酮在肠道的吸收主要以被动扩散为主,UPLC检测方法简单、灵敏度高,可用于研究染料木黄酮在Caco-2单细胞层模型上的吸收特征。
Objective To study the absorption characteristics of genistein on the Caco-2 monolayer model. Methods The genistein samples were centrifuged at high speed and the supernatant was taken. Acetonitrile - ammonium acetate aqueous solution was used as the mobile phase and testosterone was used as the internal standard. Ultra performance liquid chromatography (UPLC) was used to detect the genistein samples. The effects of time, pH and other factors on the genistein absorption were investigated. The concentration of genistein that passed through the Caco-2 monolayer was measured and the apparent permeability coefficient (Papp) was calculated. Results The uptake of genistein by Caco-2 cells was linear within 1 h. The drug uptake time was 1 h. The pH value had no significant effect on the genistein absorption. The linear range of genistein was 0.625 ~ 40μmol / L, with the precision of less than 15% during the day and the day. Papp was 1.51 × 10 5 cm / s from the apical (AP) to the basolateral (BL) and 1.84 × 10 5 cm / s from the BL to the AP, and its permeability coefficient Papp (BL-AP) / Papp (AP-BL) is 1.22. Conclusions Genistein is absorbed mainly by passive diffusion in intestinal tract. UPLC assay is simple and sensitive and can be used to study the absorption characteristics of genistein on Caco-2 single-cell layer model.