Development of a liquid chromatography/electrospray selected reaction monitoring method for the determination of organoarsenic species in marine and freshwater samples.

Cation- and anion-exchange high-performance liquid chromatography/electrospray selected reaction monitoring (HPLC/ES-SRM) methods were developed for the determination of 15 organoarsenic compounds in marine and freshwater samples. The results demonstrate that the developed HPLC/ES-SRM methods are powerful approaches for the identification of organoarsenic species in crude sample extracts. The detection limits, linearity as well as reproducibility for most of the species are comparable or even better than those measured by the HPLC/inductively coupled plasma mass spectrometry (ICPMS) technique. The qualitative analysis of the extracts shows that the developed methods allow for the identification of arsenicals which were not detectable by ICPMS. It was also demonstrated that the signal suppression caused by matrix effects means a significant limitation in the quantification of arsenicals by ES-SRM detection. This drawback is manifested especially in the case of the slightly retained species. The three sample-cleanup chromatographic methods including off-line size-exclusion, on-line reversed-phase and on-line oppositely charged ion-exchange approaches proved to be ineffective for separation of the signal-suppressive matrix from the analytes. The standard addition calibration seems to be a suitable solution for such problems.

Arsenic speciation in freshwater organisms from the river Danube in Hungary.

Total arsenic and arsenic species were determined in a range of freshwater samples (sediment, water, algae, plants, sponge, mussels, frog and fish species), collected in June 2004 from the river Danube in Hungary. Total arsenic concentrations were measured by ICPMS and arsenic species were measured in aqueous extracts of the samples by ion-exchange HPLC-ICPMS. In order to separately determine the efficiency of the extraction method and the column recovery, total arsenic concentrations in the extracts were obtained in three ways: (i) ICPMS determination after acid digestion; (ii) flow injection analysis performed directly on the extract; (iii) the sum of arsenic species eluting from the HPLC column. Extraction efficiencies were low (range 10-64%, mean 36%), but column recovery was acceptable (generally >80%) except for the fish samples, where substantial, currently unexplained, losses were observed. The dominating arsenic species in the extracts of freshwater algae were arsenosugars, whereas arsenate [As(V)] was present only as a minor constituent. On the other hand, plant extracts contained only inorganic arsenic, except for two samples which contained trace amounts of dimethylarsinate (DMA) and the tetramethylarsonium cation (TETRA). The oxo-arsenosugar-phosphate (ca. 35% of extractable arsenic) and the oxo-arsenosugar-glycerol (ca. 20%) as well as their thio-analogues (1-10%) were found in the mussel extracts, while arsenobetaine (AB) was present as a minor species only. In general, fish extracts contained only traces of arsenobetaine, and the oxo-arsenosugar-phosphate was the major arsenic compound. In addition, samples of white bream contained thio-arsenosugar-phosphate; this is the first report of a thio-arsenical in a fish sample. The frog presented an interesting arsenic speciation pattern because in addition to the major species, arsenite [As(III)] (30%) and the tetramethylarsonium cation (35%), all three intermediate methylation products, methylarsonate (MA), dimethylarsinate and trimethylarsine oxide (TMAO), and arsenate were also present. Collectively, the data indicate that arsenobetaine, the major arsenical in marine animals, is virtually absent in the freshwater animals investigated, and this represents the major difference in arsenic speciation between the two groups of organisms.

Arsenic speciation in farmed Hungarian freshwater fish.

Arsenic speciation analysis was carried out on freshwater farmed fish collected from an area with elevated groundwater arsenic concentrations in Hungary as well as from outside of the area (control samples). The arsenic species were determined by high-performance liquid chromatography-inductively coupled plasma mass spectrometry on methanol extracts of the muscle tissue from the fish. Catfish (Claries gariepinus) were raised in geothermal water where the average total arsenic concentrations were 167 (contaminated sites) and 15.1 ng As mL(-1) (control); they were all fed an artificial diet containing 2880 microg As kg(-1) total arsenic, mostly present as arsenobetaine. In the catfish, the accumulated total arsenic (2510-4720 microg As kg(-1)) was found mostly in the form of arsenobetaine suggesting that uptake of arsenic was dominated by their diet. Carp (Cyprinus carpio) were cultured in surface lakes with no significant arsenic pollution and had total arsenic concentrations ranging from 62 to 363 microg As kg(-1). The arsenic species found in the carp extracts differed markedly from those in the catfish in that no arsenobetaine was detected. Most samples of carp from the investigated sites contained low concentrations of As(III) (arsenite), As(V) (arsenate), MA (methylarsonate), and DMA (dimethylarsinate), and no other compounds were detected. The four individuals from the control site, however, all contained appreciable levels of oxo-arsenosugar-glycerol and oxo-arsenosugar-phosphate. Indeed, the oxo-arsenosugar-phosphate dominated the speciation pattern for these carp contributing about 75% of the sum of species. The contrast between these two freshwater aquaculture species regarding total arsenic and arsenic species has relevant toxicological aspects in terms of food safety.

Thio arsenosugars in freshwater mussels from the Danube in Hungary.

In contrast to the large body of data on naturally-occurring arsenic compounds in marine organisms, relatively little is known about arsenic speciation in freshwater biota. We report an investigation using HPLC-ICPMS into the arsenic compounds in five species of freshwater mussels collected from five sites from the Danube in Hungary. Total arsenic concentrations in the mussels ranged from 3.8-12.8 mg As kg(-1). The arsenic speciation patterns were broadly similar for mussels representing each of the five species and five sites, but quite different from those reported for marine mussels. The major extractable arsenicals were two oxo arsenosugars (glycerol sugar and phosphate sugar), and their thio analogues (thio glycerol sugar and thio phosphate sugar). Arsenobetaine, usually the major arsenical in marine organisms, was not a significant compound in the freshwater mussels and was detected in only three of the 11 samples. This is the first report of thio arsenosugars in freshwater biota and suggests that these compounds may be common and widespread naturally-occurring arsenicals.