HPLC-FLU法测定血清中黄藤素的含量

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目的 建立血清样品中黄藤素的高效液相色谱 -荧光检测法。方法 色谱柱为LiChrosorbSI 6 0 (5 μm)2 0cm× 4 0mmID ;流动相为二氯甲烷—甲醇—二乙胺—冰醋酸 (90∶9∶0 4∶0 5 ) ;流速为 1 0mL·min-1;荧光检测Ex=36 5nm ,Em=5 10nm。血清样品加 13 甲基小檗碱 (内标 ) ,用三氯乙酸 氯仿提取 ,水浴加温、氮气吹干 ,用二氯乙烷溶解进样。结果 黄藤素和内标的保留时间分别为 8 4min和 7 1min。黄藤素的最低检测浓度为 0 1ng·mL-1。在 0 1~ 10 0ng·mL-1有良好线性关系 (γ =0 9999)。血药浓度测定天内、天间精密度分别为 0 94%~1 85 %和 0 15 %~ 6 47%。结论 用高效液相色谱 荧光检测法测定黄藤素的血药浓度 ,有灵敏、专一和快速的优点 ,可满足药代动力学研究的需要 Objective To establish a HPLC method for the determination of fibronin in serum. Methods The column was LiChrosorbSI 60 (5 μm) 20 cm × 40 mm, the mobile phase was dichloromethane-methanol-diethylamine-glacial acetic acid (90: 9: 4: 5) -1; fluorescence detection Ex = 36 5nm, Em = 5 10nm. Serum samples plus 13 methyl berberine (internal standard), extracted with trichloroacetic acid chloroform, water bath heating, dry with nitrogen, dissolved in dichloroethane injection. Results The retention time of Ficusin and internal standard were respectively 8 4min and 7 1min. The lowest detectable concentration of palmatine was 0 1 ng · mL-1. There was a good linear relationship between 0 1 ~ 10 0 ng · mL -1 (γ = 0 9999). Intra-day precision was between 0 94% and 1 85% and 0 15% to 6 47%, respectively, for the determination of plasma concentrations. Conclusion The determination of blood plasma concentration of fluticasin by high performance liquid chromatography with fluorescence detection is sensitive, specific and rapid, which can meet the needs of pharmacokinetic studies
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