THE DETERMINATION OF ENVIRONMENTAL AND INDUSTRIAL-EXPOSURE TO HEAVY-METALS BASED ON THE QUANTITATIVE ISOLATION OF METALLOTHIONEIN FROM HUMAN FLUIDS, WITH APPLICATION OF COVALENT AFFINITY-CHROMATOGRAPHY WITH THIOL-DISULFIDE INTERCHANGE GEL AS A SOLID-PHASE EXTRACTION SUPPORT

The aim of this study was to present a new analytical method for the q uantitative determination of metallothionein (MT) proteins in human bo dy fluids and tissues, in order to determine the level of environmenta l and industrial exposure to heavy metals. For MT isolation, covalent affinity chromatography with thiol-disulphide inter-change (CAC-TDI) w as applied. Fundamentals of indirect determination of the contents of metallothionein proteins were worked out through estimation of the qua ntities of metals bound with metallothionein protein and adsorbed on c ovalent affinity chromatography gel as on solid-phase extraction suppo rt during separating process. The (CAC-TDI) gel, specially prepared, w as used as a solid phase extraction support (SPE) for preconcentration of Hg-thionein (Hg-Th), Cd-thionein (Cd-Th), Zn-thionein (Zn-Th) and Cu-thionein (Cu-Th) proteins and Hg, Cd, Zn and Cu bonded with MTs fro m water, human fluids such as: urine, human plasma, breast milk and ti ssues homogenates. (C) 1998 Elsevier Science B.V. All rights reserved.