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New metallothionein assay in Scrobicularia plana: heating effect and correlation with other biomarkers |
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| Authors: | Romero-Ruiz Antonio Alhama José Blasco Julián Gómez-Ariza José Luis López-Barea Juan |
| Affiliation: | a Department of Biochemistry and Molecular Biology, University of Córdoba, Córdoba, Spain b Andalusia Institute of Marine Sciences-CSIC, Río San Pedro Campus, Puerto Real, Cádiz, Spain c Department of Chemistry and Materials Sciences, University of Huelva, Huelva, Spain |
| Abstract: | Metallothionein (MT) and other biomarker levels were measured in Scrobicularia plana clams to assess pollution of the Guadalquivir Estuary possibly affected by metals released from Aznalcóllar pyrite mine in 1998. After optimizing reagent concentrations for monobromobimane derivatization, MT levels were quantified by reversed-phase high-performance liquid chromatography coupled to fluorescence detection (RP-HPLC-FD) in heated or unheated digestive gland extracts and compared to those obtained by differential pulse polarography (DPP). MT content assayed by RP-HPLC-FD in unheated samples was higher than that obtained by DPP and correlated better with metals and anti-oxidant activities. MT assay by RP-HPLC-FD in unheated extracts would be preferable for assessing metal pollution, due to its greater sensitivity and specificity. In addition to MT induction, glyoxalase II inhibition was well correlated with metal contents. Our results suggest that metals at the estuary do not originate from Aznalcóllar spill, but from those carried along by the river and deposited at its concave bank. |
| Keywords: | Bivalve molluscs Metals Reversed-phase high performance liquid chromatography Fluorescence detection Doñ ana National Park Guadalquivir Estuary Aznalcó llar pyrite mine |
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