A metallomic approach based on the use of size-exclusion chroma tography coupled to inductively coupled plasma-mass spectrometry (SECICP-MS) has been used to characterize the bio logical response in liver, brain, kidneys, lungs and plasma of the free-living mou se Mus spretus in polluted areas located in Donana National Park (southwest Spain) and the surroundings, mainly affected by agriculture, mining and industry activities, which are responsible for the presence of metallic contaminants. It is remarkable the high presence of Cu, Z n, Cd, As, Pb and Ni in the cytosolic extracts of different organs and plasma, especially in co ntaminated areas. In liver extracts, high intensity peaks traced by Cu, Zn, Pb and Cd at 7 kDa (matching with metallothionein I standard) are triggered by the presence of contaminants. In kidn ey, similar Cu and Cd-peaks at 7 kDa were observed but the equivalent Zn-peak was depleted by the competitiv e interactions of Cu-Cd-Zn for the active sites of these molecules. In addition, peaks traced by Cu and Zn at about 32 kDa in liver extract match with superoxide dismuta se standard (Cu,Zn-SOD), which increase in accordance to contamination. An analogous behavior was observed for a Zn,Cu-peak at about 67 kDa that can be related with the bovine serum albumin standard (Cu,Zn-BSA ) or other carrier protein such as transferrin (Cu-Tf) present in liver and plasma. Finally, low molecular mass arsenic metabolites were detec ted in mice captured in MAT site affected by mine waste.
Abstract A method for speciation of dimethylselenide (DMeSe), dimethyldiselenide (DMeDSe) and diethylselenide (DEtSe) in sediments based on a coupling between a pervaporation module, a preconcentration sorptive trap and a gas chromatograph-mass spectrometer is reported. The coupling is performed through a high pressure injection valve which allows two different operational modes: (a) analysis without preconcentration, in which analytes are directly driven from the pervaporation chamber to the injection port of the chromatograph, and (b) analysis with preconcentration in a trap, in which the analytes from the pervaporation chamber are first trapped on a Tenax minicolumn and then thermally desorbed and driven to the GC. This second approach improves the sensitivity compared to the direct coupling, reaching estimated absolute detection limits lower than 0.6 ng Se for each tested species. The method is applied to the determination of volatile organic selenium species in several sediments collected from different areas in the Southwest of Spain.
Organic and inorganic mass spectrometries were used to investigate the biochemical response of mice (Mus musculus) to inorganic arsenic exposure using liver as the target organ. The toxicological effects of trivalent inorganic arsenic after oral administration (3 mg kg−1 body weight and per day) were investigated over a period of 7 days using metallomics, metabonomics and redox proteomics approaches. Size-exclusion chromatography (SEC) with ICP-MS detection was combined with anion exchange chromatography (AEC) to characterize the biological response of the exposed mice. On the other hand, direct infusion mass spectrometry (DI-ESI-QTOF-MS) of polar and lipophilic extracts using positive and negative modes of acquisition (ESI+/ESI−) provided information about time-dependent changes in endogenous metabolites identified by Partial Least Square-Discriminant Analysis (PLS-DA). Finally, the study has been complemented with the evaluation of up/down-regulation of enzymes related to oxidative stress such as superoxide dismutase (SOD), glutathione reductase (GR), catalase (CAT) and peroxidases in connection with metal toxicity issues. The results show that the inorganic arsenic methylation in the liver may reach the saturation point upon chronic exposure to the element. On the other hand, SEC-ICP-MS coupling provided information about metal containing-proteins and metabolites related to arsenic exposure (metallomics) which has been correlated with the changes in the global metabolism (metabonomics), also considering their consequences on the redox status of protein and protein expression (redox proteomics). Our study shows that arsenic causes biochemical pathway alterations, such as energy metabolism (e.g. glycolysis, Krebs cycle), amino acid metabolism, choline metabolism and degradation of membrane phospholipids (apoptosis). This work illustrates the high reliability of the integrated use of organic mass spectrometry for the metabonomic study of biochemical effects induced by As2O3, with inorganic mass spectrometry for metallomic and speciation assessment of arsenic biomethylation in the liver of exposed mice, and redox proteomics to evaluate inhibition of enzymatic activity in different proteins such as superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR) caused by this element. In conclusion, the integration of metallomics, metabolomics and redox proteomics results provides a more comprehensive evaluation about the biological response in experiments dealing with exposure to toxic metals.