Characterisation of metal-binding biomolecules in the clam Chamelea gallina by bidimensional liquid chromatography with in series UV and ICP-MS detection
2011
Vitoulová, Eva | García Barrera, Tamara | Gómez-Ariza, José Luis | Fišera, Miroslav
The presence of metal-binding biomolecules has been studied in Chamelea gallina, a bioindicator used for environmental contamination monitoring and very popular for human consumption in the Atlantic southwest coast of Spain. This area is affected by metal pollution from mining activities, which can modify biomolecules expression in this bivalve. Total content of elements was determined by ICP-MS and revealed a remarked presence of Fe, Zn, Cu, As and Mn. A metallomics approach has been optimised for this mollusc using size-exclusion chromatography on column Superdex 30 pg HiLoad 26/60 with in series UV and ICP-MS detection. At least four fractions with molecular weight in the range 1540 to 415 Da were observed with UV detection, but the ICP-MS chromatogram showed the presence of metals of interest only in the first two fractions. The apparent molecular weights of these metal-containing fractions were from 1325 to 764 Da. The fractions containing metals compounds were collected and lyophilised for further purification of reconstituted extracts with a second orthogonal chromatographic separation using reverse phase (RP) HPLC with ICP-MS detection. Several peaks were obtained in this second dimension separation which allows the isolation of As-, Cu- and Zn-containing biomolecules.
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