Abstract
AbstractThe speciation of trace element, in clinical and biological samples has become a focusing discipline, which provides significant information for biochemist to explore the mechanism of availability, cycling, transformation, bioaccumulation and final fate of these “biometals”. In view of the important role of bio-molecules played in the biological systems, development of reliable analytical method aiming at elucidating the structural and quantitative information about these materials is highly demanded.For the purpose of characterizing biometals which bind with various bio-molecules, a hyphenation technique coupling size exclusion chromatographic separation and ICP-MS detection was explored. Various parameters of SEC, such as mobile phase flow rate and sample volume on the effect of separation efficiency were investigated. Following the optimized separation condition, trace amount of Cu, Zn, Cd, Pb and Co in human serum were found bound with the biomolecule of M.W.~6,500 Da and a broad As-containing peak corresponding to M.W.~1,500 Da was observed on SEC-ICPMS chromatograph. The identification of the species was prelimernarily investigated with a molecular specific ESI-MS technique.In this study an on-line SEC-ID-ICPMS system for the quantitative determination of binding and non-binding species of Cu in human serum has also been developed. The accuracy of this hyphenation system for the determination of total concentration of Cu in human serum was verified with the use of NIST SRM 1598 and 311089. With this SEC-ID-ICP-MS system, the presence of the Cu-biomolecule of molecular weight of about 60,000 Da, was found to be about 90% of the total Cu concentration, which is in good agreement with that reported in the literature.