超滤法测定天麻素与不同血浆的蛋白结合率

Determination of protein binding rate of gastrodin with different plasma by ultrafifi ltration

目的建立测定天麻素与小鼠、大鼠、家兔的血浆蛋白结合率的实验方法,并计算天麻素与不同血浆的蛋白结合率。方法采用HPLC法测定天麻素的含量,Agilent TC-C18(250 mm×4.6 mm,5μm)色谱柱,流动相为甲醇:水(10:90,v/v),体积流量为1.0 mL/min,柱温为25℃,检测波长为220 nm。通过超滤法比较天麻素与小鼠、大鼠、家兔的血浆蛋白结合率。结果天麻素与小鼠血浆在5、20、50μg/m L浓度下的蛋白结合率为36.18%±0.54%、35.93%±0.48%、36.26%±0.25%;与大鼠血浆在5、20、50μg/m L浓度下的蛋白结合率为35.86%±0.27%、36.11%±0.36%、36.57%±0.39%;与家兔血浆在5、20、50μg/m L浓度下的蛋白结合率为37.00%±0.22%、36.90%±0.17%、37.15%±0.31%。结论天麻素与小鼠、大鼠、家兔血浆均属于低强度结合,无差异性和浓度依赖性。

Objective To establish an experimental method for determining the plasma protein binding rate of gastrodin to mice, rats and rabbits, and the protein binding rate of gastrodin to different plasma was calculated.Methods The content of gastrodin was determined by HPLC on the Agilent TC-C18（250 mm×4.6 mm, 5 μm）column, the mobile phase was methanol： water（10：90, v/v）, the volume flow rate was 1.0 mL/min, the column temperature was 25, and the detection wavelength was 220 nm. The plasma protein binding rate of gastrodin to mice, rats and rabbits were compared by ultrafi ltration method. Results The protein binding rate of gastrodin to mouse plasma at 5, 20, 50 μg/mL were 36.18%±0.54%, 35.93%±0.48%, 36.26%±0.25%, respectively; and to rat plasma were 35.86%±0.27%, 36.11%±0.36%, 36.57%±0.39%, respectively; and to rabbit plasma were 37.00%±0.22%,35.90%±0.17%, 37.15%±0.31%, respectively. Conclusion The plasma of gastrodin in mice, rats and rabbits were low intensity binding, no difference and concentration dependence.