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采用凝胶色谱、离子交换色谱以及反相高效液相色谱法,将尿毒症患者的尿样和血样以及正常人的尿样和血样进行了多级分离和比较.通过凝胶色谱法,从正常人的尿液以及尿毒症血清和尿液中分离出相对于正常人血清具有异常高浓度的A,B2个中分子组分.不同试样来源的A峰中分子物的离子交换色谱分离结果证明,A-3亚峰中分子物,是尿毒症患者因肾衰难以通过尿液将其排除故而滞留在血清内的重要成分,正常人却可以通过尿液将其排于体外.将来自尿毒症血清及正常人尿液的A-3亚峰中分子物经脱盐处理后,进行了基质辅助激光解吸电离-飞行时间质谱分析,证明A-3组分由分子量分别为839,873,1007,1106,1680,2015的6种化合物组成.最后,采用反相高压液相色谱法对A-3亚峰中分子组分进行了进一步分离,得到了6个只含单一中分子化合物成分的组分.至此,A-3 组分内的中分子物得到了彻底的分离和纯化,为进一步确定其结构以及病理生理作用奠定了基础.
The urinary and blood samples from patients with uremia and the urine and blood samples from normal people were separated and compared by gel filtration, ion exchange chromatography and reversed-phase high performance liquid chromatography (HPLC) Human urine and uremic serum and urine were separated from normal human serum with abnormally high concentrations of A, B2 in the molecular components of different sample source A peak molecular ion exchange chromatography results show that , A-3 Yafeng molecular objects, uremia patients due to renal failure is difficult to remove it through the urine and remain in the serum of the important elements, but normal urine but it can be excluded from the body will be from uremia Serum and normal urine of A-3 sub-peak in the desalting of molecular compounds, the matrix-assisted laser desorption ionization-time of flight mass spectrometry analysis showed that A-3 components from the molecular weight of 839,873,1007,1106,1680 , 2015. Finally, the molecular components in the A-3 sub peak were further separated by RP-HPLC and six components were obtained, which contained only a single intermediate molecular compound.Thus, A-3 components in the middle of the matter derived Thorough separation and purification, to further determine the basis for its construction and its pathophysiological role.