The aim of this study was to present a new analytical method for the quantitative determination of metallothioneins (MT) protein in human body fluids and tissues, in order to determine the level of environmental and industrial exposition to heavy metals. For MTs isolation, covalent affinity chromatography with thiol-disuphide interchange (CAC-TDI) was applied, which is a modern technique of separation with high affinity, good repeatability, and reproducibility, allowing specific isolation of the thiolproteins and metallothiolproteins. Fundamentals of indirect determination of the contents of metallothioneins protein were worked out through estimation of the quantities of metals bound with metallothionein protein and adsorbed on covalent affinity chromatography gel as a solid-phase extraction support during the separation process.
The (CAC-TDI) gel, specially prepared, was used as a solid-phase extraction support (SP ) 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 from spiked water, urine, human plasma, breast milk, and tissues homogenates.