HPLC-超滤法测定羟基酪醇与常氧及缺氧大鼠血浆蛋白的结合率

Determination of Plasma Protein Binding Rate of Hydroxytyrosol in Normoxic and Hypoxic Rat by HPLCUltrafiltration Method

目的建立羟基酪醇血浆蛋白结合率的测定方法,比较常氧及缺氧大鼠血浆中羟基酪醇蛋白结合率的异同。方法采用HPLC-超滤法测定羟基酪醇与常氧及缺氧大鼠血浆的蛋白结合率。结果当大鼠血浆中羟基酪醇浓度为3,6,12μg·mL^(–1)时,羟基酪醇与常氧大鼠血浆的蛋白结合率分别为(17.23±1.11)%,(16.88±1.37)%和(16.70±0.98)%,与缺氧大鼠血浆的蛋白结合率分别为(12.31±1.79)%,(12.75±1.20)%和(13.42±1.98)%,两者比较差异有统计学意义(P<0.01)。结论羟基酪醇与常氧大鼠血浆蛋白属于低强度结合,且在3~12μg·mL^(–1)内,不具浓度依赖性,而羟基酪醇与缺氧大鼠血浆蛋白结合强度明显低于与常氧大鼠血浆蛋白的结合。

OBJECTIVE To establish a method for determination of plasma protein binding rate of hydroxytyrosol,and compare the difference of plasma protein binding rate in normoxic and hypoxic rats.METHODS The protein binding rate of hydroxytyrosol in normoxic and hypoxic rats plasma was determined by HPLC-ultrafiltration.RESULTS When the concentration of hydroxytyrosol in rat plasma was 3,6 and 12μg·mL^(-1),the plasma protein binding rates of hydroxytyrosol with normoxic rats were(17.23±1.11)%,(16.88±1.37)%and(16.70±0.98)%,and the plasma protein binding rates with hypoxic rats were(12.31±1.79)%,(12.75±1.20)%and(13.42±1.98)%.The difference of the plasma protein binding rate was significant between the normoxic and hypoxic rats(P<0.01).CONCLUSION The binding strength of hydroxytyrosol to plasma protein of normoxic rats is low and does not show concentration dependence within the concentration range of 3–12μg·mL^(-1).The binding strength of hydroxytyrosol to plasma protein of hypoxic rats is significantly lower than that of hydroxytyrosol to plasma protein of normoxic rats.