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目的:建立血浆样品中丹酚酸A浓度的分析方法,并测定丹酚酸A的血浆蛋白结合率,为进一步展开丹酚酸A的代谢和药动学研究及指导临床用药提供参考。方法:采用超滤法和HPLC测定丹酚酸A在牛血清白蛋白(BSA),大鼠血浆,新西兰兔血浆和比格犬血浆中的蛋白结合率。结果:丹酚酸A在0.01～2.5 mg.L-1线性良好,标准曲线方程为Y=0.807 3X+0.013 2(r=0.999 4)。丹酚酸A与BSA,大鼠血浆,新西兰兔血浆和比格犬血浆在5.0,50.0,100.0 mg.L-1的含药血浆中其平均蛋白结合率分别为(99.79±0.02)%,(99.79±0.03)%,(99.73±0.06)%,(99.81±0.03)%。结论:所建立的方法灵敏度高,专属性和重复性好,操作简单,能够满足定量分析测试要求。丹酚酸A与血浆蛋白有很强的结合,且在已考察的血药浓度范围内其血浆蛋白结合率无明显浓度依赖性和种属差异性。
OBJECTIVE: To establish a method for the determination of salvianolic acid A concentration in plasma samples and to determine the plasma protein binding rate of salvianolic acid A in order to provide a reference for further research on the metabolism and pharmacokinetics of salvianolic acid A and for guiding the clinical use. Methods: The protein binding of salvianolic acid A in plasma of bovine serum albumin (BSA), rat plasma, New Zealand rabbit plasma and beagle dog was determined by ultrafiltration and HPLC. Results: The linear range of salvianolic acid A was 0.01-2.5 mg.L-1. The standard curve equation was Y = 0.807 3X + 0.013 2 (r = 0.999 4). The average protein binding rates of salvianolic acid A to BSA, plasma of rat, plasma of New Zealand rabbit and plasma of beagle dog were (99.79 ± 0.02)% in 5.0, 50.0 and 100.0 mg.L-1 plasma respectively, ( 99.79 ± 0.03)%, (99.73 ± 0.06)%, (99.81 ± 0.03)%. Conclusion: The established method has high sensitivity, specificity and repeatability, simple operation and can meet the quantitative analysis and testing requirements. Salvianolic acid A has a strong binding to plasma proteins and has no significant concentration-dependent and species-specific plasma protein binding within the range of plasma concentrations examined.