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目的建立芒果苷血浆蛋白结合率的测定方法,比较芒果苷在糖尿病大鼠及正常大鼠血浆中蛋白结合率的异同。方法采用血浆平衡透析法,LC-MS/MS法测定透析袋两侧溶液中芒果苷的质量浓度,分别计算其在糖尿病大鼠和正常大鼠血浆中的蛋白结合率。结果血浆和磷酸盐缓冲液(PBS)中,芒果苷分别在200~3 000ng·mL~(-1)(r=0.998 3),50~2 000 ng·mL~(-1)(r=0.997 9)范围内线性关系良好,准确度均在94.7%~108.6%之间,日内、日间相对标准偏差(RSD)均小于15%。芒果苷质量浓度为100,200,500 ng·mL~(-1)时,糖尿病大鼠的平均血浆蛋白结合率分别为:(84.5±1.4)%、(82.6±1.4)%、(82.4±1.7)%;正常大鼠的平均血浆蛋白结合率分别为:(65.5±0.6)%、(66.6±1.0)%、(68.4±0.7)%,两者比较差异有统计学意义(P<0.01)。结论 LC-MS/MS法可用于芒果苷血浆蛋白结合率的测定。芒果苷与糖尿病大鼠的血浆蛋白结合率属于高度结合(>80%),具有非浓度依赖性,且明显高于正常大鼠。
Objective To establish a method for the determination of plasma protein binding of mangiferin and to compare the protein binding rate of mangiferin in plasma of diabetic rats and normal rats. Methods The concentration of mangiferin in solution on both sides of dialysis bag was measured by plasma equilibrium dialysis and LC-MS / MS. The protein binding rate in plasma of diabetic rats and normal rats were calculated respectively. Results In the plasma and phosphate buffered saline (PBS), the concentrations of mangiferin were in the ranges of 200-3 000 ng · mL -1 (r = 0.998 3) and 50-2 000 ng · mL -1 (r = 0.997 9) ranged from 94.7% to 108.6%. The relative standard deviations (RSDs) were less than 15% in the day and the day. The mean plasma protein binding rates of diabetic rats at the concentrations of 100, 200 and 500 ng · mL -1 were (84.5 ± 1.4)%, (82.6 ± 1.4) and (82.4 ± 1.7)%, respectively The mean plasma protein binding rate in rats was (65.5 ± 0.6)%, (66.6 ± 1.0)% and (68.4 ± 0.7)%, respectively. There was significant difference between the two groups (P <0.01). Conclusion LC-MS / MS method can be used to determine the plasma protein binding rate of mangiferin. The plasma protein binding rate between mangiferin and diabetic rats is highly binding (> 80%), which is non-concentration-dependent and significantly higher than that of normal rats.