阴离子交换树脂固相萃取分离全血中血红蛋白的研究

Investigations on the Isolation of Hemoglobin from Human Whole Blood by Solid Phase Extraction with 330 Anion Exchange Resin

以330阴离子交换树脂为吸附材料建立了固相萃取分离血红蛋白的方法。利用血红蛋白和树脂之间的疏水作用力将血红蛋白吸附到树脂上,以Tris-HCl缓冲液(pH=8.9)为洗脱剂回收血红蛋白。考察了溶液pH值、吸附时间、离子强度、洗脱剂的种类及其酸度等对分离纯化效率的影响。在最优实验条件下,树脂对血红蛋白的吸附率和洗脱率分别为87%和70%,吸附容量为42.9μg/mg。吸收光谱和SDS-PAGE凝胶电泳证明,该方法可有效地从人全血中分离出纯度较高的血红蛋白。

A solid-phase extraction procedure for the isolation of hemoglobin from human whole blood was developed with 330 anion exchange resin as sorbent. Hemoglobin was adsorbed onto the resin surface via hydrophobic interaction which was afterwards collected by stripping with Tris-HCl buffer （pH=8. 9） as eluent. Some of the experimental parameters, including sample pH, adsorption time, ionic strength and stripping reagent, governing the sorption and desorption of hemoglobin were carefully investigated. A sorption efficiency of 87% and a recovery of 70% were achieved along with a sorption capacity of 42.9 μg/mg for hemoglobin. The practical applicability of this system was demonstrated by processing of human whole blood for the isolation of hemoglobin, and satisfactory results were obtained by assays with UV-Vis spectroscopy as well as SDS-PAGE.