平衡透析法结合HPLC测定去氢骆驼蓬碱衍生物(DH-004)在大鼠血浆的蛋白结合率

Protein binding rate of harmine derivative(DH-004)in the rats plasma determined by equilibrium dialysis combined with HPLC

目的研究去氢骆驼蓬碱衍生物(DH-004)在大鼠血浆的蛋白结合率。方法Wistar大鼠腹主动脉采血,肝素抗凝,4℃下以3.5×10^(3) r·min^(-1)离心15 min后分离血浆。以替硝唑为内标,用平衡透析法处理样品,用高效液相色谱(HPLC)法测定透析袋内外溶液中DH-004的浓度,计算蛋白结合率。结果DH-004于大鼠血浆中在1.05,2.10,5.26μg·mL^(-1)下的血浆蛋白结合率分别为(89.96±0.89)%,(90.79±0.85)%和(91.39±1.00)%。结论DH-004与大鼠血浆蛋白有较强的结合作用,属于高蛋白结合型药物。

Objective To determine the plasma protein binding rate of harmine derivative(DH-004).Methods The abdominal aorta of blood was collected from Wistar rats,and heparin sodium was used as anticoagulant.The plasma was separated at 4℃by centrifugation at 3.5×10^(3) r·min^(-1) for 15 min.Tinidazole was regarded as internal standard substance,and equilibrium dialysis method was applied to pretreat plasma sample.The concentrations of DH-004 inside and outside of dialysis bag were measured by high performance liquid chromatography(HPLC).Protein binding rate was calculated.Results The plamsa protein binding rates of DH-004 in rat plasma at 1.05,2.10,5.26μg·mL^(-1) were(89.96±0.89)%,(90.79±0.85)%and(91.39±1.00)%,respectively.Conclusion DH-004 have strong binding effect on rat plasma protein,and belong to high protein binding drugs.