原子吸收光谱法测定血清中不同化学形态的铜、铁、锌

融合分子生物学技术与原子吸收光谱对血清中元素铜、铁和锌的化学形态进行研究。用60%乙醇低温(4℃)沉淀血清蛋白的方法将血清中的铜,铁,锌分为结合态和非结合态,原子吸收分光光度计分别测定血清中Cu,Fe和Zn三种元素的总量及非结合态含量,通过减差法求出血清中结合态元素含量,从而建立了Cu,Fe和Zn元素这两种化学形态的分离分析方法,并讨论了有关的实验条件。该方法铜、铁和锌的检出限分别为9·84×10-3μg·mL-1,2·76×10-2μg·mL-1,1·06×10-3μg·mL-1,相对标准偏差为0·30%～2·31%,回收率为95·0%～104·0%。该法已应用于SD大鼠血清中Cu,Fe和Zn三种元素不同化学形态的测量。

Speciation Analysis of Trace Elements Cu,Fe and Zn in Serum by Flame Atomic Absorvtion Spectrophotometry

Since biological functions of the elements are generally different, depending on their chemical forms, chemical speciation analysis is really important in metallomics research. Thus, multielement analysis and chemical speciation of the elements in serum were carried out in the present work. A hyphenated technique was developed for high-throughput speciation analysis of the copper, iron and zinc in serum by molecular biology technology and flame atomic absorption spectrophotometry (AAS). Here, Cu, Fe and Zn in serum were classifyied as the forms of combination and non-combination. The serum protein was precipitated by 60% concentration of ethanol under hypothermy. The forms of combination of Cu, Fe and Zn in serum which combined with proteins were in precipitations, and the forms of non-combination of Cu, Fe and Zn in serum, which were free ions, were in supernatant. The total amount of Cu, Fe and Zn in serum and the amount of the forms of non-combination of Cu, Fe and Zn were analyzed by AAS. The amount of the forms of combination of Cu, Fe and Zn was obtained by calculation. The detection limit of Cu in serum by the method is around and 9.84 x 10(-3) microg x mL(-1). For Fe and Zn, the detection limit is about 2.76 x 10(-2) microg x mL(-1) and 1.06 x 10(-3) microg x mL(-1), respectively. The percentage recovery of trace elements Cu, Fe and Zn by the proposed procedure is in the range 95.0%-101.0%, 95.0%-102.0% and 95.0%-103.0%, respectively. The relative standard deviation (RSD) of trace elements Cu, Fe and Zn in the serum is in the range 1.88%-2.26%, 0.56%-1.59% and 0.34%-1.36%, respectively. Speciation of trace elements Cu, Fe and Zn in the serum of SD rat were analyzed by the method.