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目的:研究白头翁皂苷D与不同血浆蛋白的结合率。方法:采用HPLC与平衡透析法相结合的方法测定白头翁皂苷D与牛血清、人血浆及大鼠血浆中蛋白的结合率,COSMOSIL 5C18-MS-Ⅱ色谱柱(4.6 mm×250 mm,5μm),流动相甲醇-水-甲酸(80∶20∶0.02),检测波长203 nm。结果:白头翁皂苷D质量浓度为0.06,0.11,0.25 g·L-1时与牛血清白蛋白的结合率分别为(78.45±0.89)%,(77.61±1.14)%,(77.16±0.29)%,与人血浆蛋白的结合率分别为(68.91±0.54)%,(67.68±0.87)%,(67.88±0.71)%,与大鼠血浆蛋白的结合率分别为(78.15±0.76)%,(78.61±0.97)%,(78.24±0.93)%。结论:建立的白头翁皂苷D蛋白结合率测定方法简便、稳定、可靠。白头翁皂苷D是一种中等程度的蛋白结合药物,结合率不具有质量浓度依赖性。
Objective: To study the binding rate of Pulsatilla saponin D to different plasma proteins. Methods: The binding rate of Pulsatilla saponin D to bovine serum, human plasma and rat plasma was determined by a combination of HPLC and dialysis. The COSMOSIL 5C18-MS-Ⅱ column (4.6 mm × 250 mm, 5 μm) Methanol - water - formic acid (80:20:0.02), detection wavelength of 203 nm. Results: The binding rates of Pulsatilla saponin D to bovine serum albumin were (0.06,0.11,0.25 g · L-1), (77.41 ± 1.14)%, (77.16 ± 0.29)% and The binding rates to human plasma proteins were (68.91 ± 0.54)%, (67.68 ± 0.87)% and (67.88 ± 0.71)%, respectively. The binding rates to plasma proteins were (78.15 ± 0.76)% and 0.97)%, (78.24 ± 0.93)%. Conclusion: The established method for the determination of the binding rate of Pulsatilla saponin D protein is simple, stable and reliable. Pulsatilla saponin D is a moderate degree of protein binding drugs, the binding rate does not have a concentration-dependent.