Arsenic speciation in edible alga samples by microwave-assisted extraction and high performance liquid chromatography coupled to atomic fluorescence spectrometry.

Twelve commercially available edible marine algae from France, Japan and Spain and the certified reference material (CRM) NIES No. 9 Sargassum fulvellum were analyzed for total arsenic and arsenic species. Total arsenic concentrations were determined by inductively coupled plasma atomic emission spectrometry (ICP-AES) after microwave digestion and ranged from 23 to 126 µg g(-1). Arsenic species in alga samples were extracted with deionized water by microwave-assisted extraction and showed extraction efficiencies from 49 to 98%, in terms of total arsenic. The presence of eleven arsenic species was studied by high performance liquid chromatography-ultraviolet photo-oxidation-hydride generation atomic-fluorescence spectrometry (HPLC-(UV)-HG-AFS) developed methods, using both anion and cation exchange chromatography. Glycerol and phosphate sugars were found in all alga samples analyzed, at concentrations between 0.11 and 22 µg g(-1), whereas sulfonate and sulfate sugars were only detected in three of them (0.6-7.2 µg g(-1)). Regarding arsenic toxic species, low concentration levels of dimethylarsinic acid (DMA) (<0.9 µg g(-1)) and generally high arsenate (As(V)) concentrations (up to 77 µg g(-1)) were found in most of the algae studied. The results obtained are of interest to highlight the need to perform speciation analysis and to introduce appropriate legislation to limit toxic arsenic species content in these food products.

Stability of total selenium and selenium species in lyophilised oysters and in their enzymatic extracts.

To obtain reliable information on speciation analysis it is necessary to previously evaluate the stability of the species in the sample of interest. Furthermore, in those cases in which sample treatment to extract the species is time-consuming, an evaluation of how to maintain species integrity in the extracts is paramount. Thus, the present paper reports the stability of total Se, SeMet and TMSe+ in freeze-dried oyster and in the enzymatic extracts stored in Pyrex and polyethylene containers at different temperatures (-18, 4 and 20 degrees C). Total selenium determinations and Se speciation were carried out by HG-AAS after acid digestion in a microwave oven and by on-line coupling of cation exchange HPLC-ICP-MS after enzymatic hydrolysis, respectively. The results obtained for the freeze-dried sample showed that total Se and the selenium species evaluated are stable for at least 12 months, under all the conditions tested. However, Se species in the enzymatic extracts are only stable for 10 days if stored at 4 degrees C in Pyrex containers. These results show that the extracts do not necessarily have to be analysed just after sample treatment.

Selenium speciation in animal tissues after enzymatic digestion by high-performance liquid chromatography coupled to inductively coupled plasma mass spectrometry.

A procedure is described for the enzymatic digestion of tuna and mussel samples that allows the determination of selenium species by high-performance liquid chromatography in conjunction with inductively coupled plasma mass spectrometry. The species were extracted by two-step enzymatic hydrolysis with a non-specific protease (subtilisin). The selenium species were separated on a Spherisorb 5 ODS/AMINO column using two different chromatographic conditions, namely phosphate buffers at pH 2.8 and pH 6.0 as mobile phases. The method determines organic (trimethylselenonium, selenocystine, selenomethionine and selenoethionine) and inorganic selenium species (selenite and selenate), but only organic selenium species were found in the samples. The sum of identified selenium species in the sample was about 30% of the total selenium present in the enzymatic extract despite the fact that recoveries of total hydrolysed selenium were 93-102%. Trimethylselenonium ion and selenomethionine were found in both tuna and mussel samples and an unknown selenium species was also found in tuna samples.

Determination of selenium species in human urine by high performance liquid chromatography and inductively coupled plasma mass spectrometry.

A method developed to determine organic and inorganic selenium species in human urine samples is presented in detail. After a simple sample treatment based on elimination of non-charged organic compounds, selenium species were separated by high performance liquid chromatography (HPLC) on a Spherisorb 5 ODS/AMINO column using two different chromatographic conditions: phosphate buffers at pH 2.8 and 6.0. Detection was carried out using an on-line inductively coupled plasma mass spectrometer (ICP-MS). Trimethylselenonium ion and two unknown selenium species in urine samples were found. Selenium species were shown to have stability problems, with the maximum allowed storage time of 1 week.