F1013人血浆蛋白结合率测定及其影响因素

Study on the Determination of Human Plasma Protein Binding Ratio of the Caspase Inhibitor F1013 and Its Influence Factors

目的测定新Caspase抑制药F1013的人血浆蛋白结合率,并考察影响测定的因素。方法采用高效液相色谱法测定不同条件下超滤分离的游离药物浓度。结果 amicon Ultra-0.5超滤装置(Millipore)操作性较强,可较好地运用于血浆蛋白结合率研究;随离心速率、超滤时间递增,人血浆游离药物浓度逐渐增高,直至稳定;随温孵时间递增,人血浆游离药物浓度逐渐减小,直至稳定。采用超滤法测定F1013人血浆蛋白结合率过程中,离心速率、超滤时间和温孵时间均对结果产生重要影响。经过优化和筛选,最佳超滤条件为采用超滤法测定F1013人血浆蛋白结合率,将含药血浆于37℃水浴温孵1 h,置Amicon Ultra-0.5超滤装置,14 000 r·min-1离心15 min。结论 HPLC法测定回收率、精密度均符合要求,具较好的可行性。超滤法测F1013平均人血浆蛋白结合率约为52%。

Objective The ultrafiltration was employed to determine the human plasma protein binding ratio of a novel Caspase inhibitor F1013.The influence factors of this determination were investigated.Methods The unbound fractions in plasma were separated with ultrafiltration equipment under different conditions and were determined with high performance liquid chromatography（HPLC）.Results The Amicon Ultra-0.5 equipment was suitable for researching the plasma protein binding ratio of F1013.The plasma protein binding ratio of F1013 was increased with the increasing of rotation speed and centrifuge time,and it was decreased with prolongated incubation time.Conclusion Human plasma protein binding ratio of F1013 was influenced by rotation speed,centrifuge time and incubation time.When the ultrafiltration was applied to study the human plasma protein binding ratio of F1013,the best ultrafiltration conditions are that the human plasma spiked with F1013 was incubated in the 37 ℃ water for 1 h,and then put into the Amicon Ultra-0.5 equipment,to centrifuge at 14 000 r·min-1 for 15 min.The method has satisfying recovery,precision,and a good feasibility.