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目的建立药根碱、巴马汀、小檗碱的LC-MS/MS测定法,研究其在Caco-2细胞上的摄取特性。方法色谱条件:Phe-nomenexluna C18柱(4.6mm×250mm,5μm);乙腈-水(4mmol·L-1醋酸铵和0.08%甲酸)为流动相,梯度洗脱;质谱条件:采用电喷雾离子源(ESI),正离子检测,MRM模式。实验考察药物浓度和P-糖蛋白(P-glycoprotein,P-gp)抑制剂环孢素A和维拉帕米对药根碱、巴马汀、小檗碱在Caco-2细胞上摄取的影响。结果药根碱、小檗碱在0.0015～0.30μmol.L-1(r=0.9998、r=1.0000),巴马汀在0.0014～0.28μmol·L-1(r=0.9998)内均呈良好的线性关系;药根碱、巴马汀、小檗碱在Caco-2细胞的摄取在一定范围内呈浓度依赖性,表现为被动扩散;P-gp抑制剂环孢素A和维拉帕米可显著增加药根碱、巴马汀、小檗碱在Caco-2细胞上的摄取。结论该法快速、灵敏、简单。药根碱、巴马汀、小檗碱在Caco-2细胞的摄取存在P-gp的参与,药根碱、巴马汀和小檗碱是P-gp的底物。
OBJECTIVE To establish LC-MS / MS determination of jatrorrhizine, palmatine and berberine and study their uptake on Caco-2 cells. Methods Chromatographic conditions: Phe-nomenexluna C18 column (4.6mm × 250mm, 5μm); acetonitrile-water (4mmol·L-1 ammonium acetate and 0.08% formic acid) as mobile phase and gradient elution; (ESI), positive ion detection, MRM mode. Effects of drug concentration and P-glycoprotein (P-gp) inhibitor cyclosporine A and verapamil on uptake of jatrorrhizine, palmatine and berberine in Caco-2 cells were investigated experimentally . Results The pharmacokinetics of jatrorrhizine and berberine showed good linearity in the range of 0.0015-0.30μmol.L-1 (r = 0.9998, r = 1.0000) and palmatine in the range of 0.0014-0.28μmol·L-1 (r = 0.9998) (P <0.05). The uptake of jatrorrhizine, palmatine and berberine in Caco-2 cells in a concentration-dependent manner showed passive diffusion. P-gp inhibitor cyclosporine A and verapamil were significantly Increased uptake of jatrorrhizine, palmatine and berberine on Caco-2 cells. Conclusion The method is fast, sensitive and simple. Jargonine, palmatine, and berberine are involved in the uptake of C-2p in P-gp, and jatrorrhizine, palmatine and berberine are substrates for P-gp.