Determination of selenium-containing species, including nanoparticles, in selenium-enriched Lingzhi mushrooms.

Mushrooms are considered a valuable food source due to their high protein and fibre and low fat content, among the other health benefits of their consumption. Selenium is an essential nutrient and is renowned for its chemo-preventative properties. In this study, batches of selenium-enriched Lingzhi mushrooms were prepared by growing mycelium and fruit in substrates containing various concentrations of sodium selenite. The mushroom fruit accumulated low levels of selenium with selenomethionine being the most abundant form in all enriched samples. Conversely, the mycelium showed significant selenium accumulation but relatively low proportions of selenomethionine. The red colour of the selenium-enriched mycelia indicated the probable presence of selenium nanoparticles, which was confirmed by single-particle inductively coupled plasma-mass spectrometry. Mean particle diameters of 90-120 nm were observed, with size distributions of 60-250 nm. Additional analysis with transmission electron microscopy confirmed this size distribution and showed that the biogenic selenium nanoparticles were roughly spherical in shape and contained elemental selenium.

82Se Metabolically-Labeled Yeast as a Matrix-Matched Isotope Dilution Standard for Quantification of Selenomethionine.

Selenized yeast is commonly used as a highly bioavailable source of selenium in dietary supplements and feed additives and is used in research settings in various disciplines due to the large number of selenium-containing metabolites formed during growth. With the selenomethionine being the major form of selenium present in selenized yeasts, its accurate quantitation is essential, however, values are frequently underestimated due to the costly and time-consuming hydrolysis-based sample preparation required to release the selenoamino acid from proteins for analysis. The National Research Council Canada has developed an 82-Se-enriched selenized yeast Certified Reference Material, SEEY-1 (DOI: 10.4224/crm.2023.seey-1) intended to be used as a matrix-matched spike material for isotope dilution analysis of selenized yeasts. The total selenium and selenomethionine contents of SEEY-1 were determined to be 322.1 ± 4.8 mg/kg (k = 2) and 635.6 ± 16.8 mg/kg (k = 2), respectively. Here we present results on the preparation of the 82-Se-enriched yeast, the certification process, and provide an example of the use of SEEY-1 as a matrix-matched spike for the analysis of selenomethionine in a sample of selenized yeast. We demonstrate here that SEEY-1 is able to compensate for the partial digestion of yeast proteins and provide reliable analytical data on Se amino acid content in under an hour instead of the 16 hours required for conventional complete acid hydrolysis.

Compilation of selenium metabolite data in selenized yeasts.

Selenium-enriched yeast has long been recognized as an important nutritional source of selenium and studies have suggested that supplementation with this material provides chemo-preventative benefits beyond those observed for selenomethionine supplementation, despite the fact that selenomethionine accounts for 60-84% of the total selenium in selenized yeasts. There is much ongoing research into the characterization of the species comprising the remaining 16-40% of the selenium, with nearly 100 unique selenium-containing metabolites identified in aqueous extracts of selenized yeasts (Saccharomyces cerevisiae). Herein, we discuss the analytical approaches involved in the identification and quantification of these metabolites, and present a recently created online database (DOI: 10.4224/40001921) of reported selenium species along with chemical structures and unique mass spectral features.

Determination of selenocyanate, selenate, and selenite in mining wastewater by GC-MS using sequential derivatization and extraction.

Selenium speciation analysis is usually carried out using complex hyphenated analytical systems such as LC-ICP-MS. Here we present a novel selenium speciation approach based on a sequential derivatization and extraction combined with gas chromatography mass spectrometry for the simultaneous determination of selenite, selenate, and selenocyanate in aqueous mine wastewater samples. Selenocyanate was derivatized with triethyloxonium tetrafluoroborate to ethylselenocyanate, which was extracted into chloroform, following which the sample was split into two aliquots. One aliquot was acidified and 3,5-bis(trifluoromethyl)-o-phenylenediamine was used for the novel derivatization of selenite to 4,6-bis(trifluoromethyl)-2,1,3-benzoselenadiazole, for the determination of selenite. For the second aliquot, concentrated hydrochloric acid was added along with 4-nitro-o-phenylenediamine to simultaneously reduce selenate to selenite and derivatize the combined "selenite + selenate" fraction to 5-nitro-2,1,3-benzoselenadiazole. The benzoselenadiazoles were extracted with chloroform and all extracts were combined for GC-MS analysis. Low ng g-1 detection limits were reported for all three species. The method is unhindered by concentrations of chloride and sulphate up to 3%, as well as nitrate concentrations up to 3% for selenocyanate and selenite analysis, with minor losses in sensitivity for selenate up to 100 ppm nitrate, making the method particularly suitable for aqueous mine waste characterization. Quantitative trace selenium speciation was achieved using cost-effective materials and apparatus on a simple-to-operate benchtop instrument. The novel methodology was tested on gold mine wastewater samples; comparing to total selenium, a 63-149% recovery as the sum of species was observed. Additionally, this novel speciation approach was compared to LC-ICP-MS based selenium speciation and a reasonable agreement was found in the species distribution.

Selective Gas Chromatography Mass Spectrometry Method for Ultratrace Detection of Selenocyanate.

The recent interest in the determination of selenocyanate (SeCN-) in wastewater systems has spurred the development of analytical methods for its determination at the ultratrace level. Since most of the current procedures require complex and costly instrumental configurations, we have developed a simple and rapid gas chromatography tandem mass spectrometry (GC/MS/MS) method able to detect SeCN- in water samples with a LOD of 0.1 ng/g Se. A 1 mL volume of aqueous sample was buffered with sodium bicarbonate and treated with triethyloxonium tetrafluoroborate for conversion of the analyte into volatile EtSeCN. The derivatization yield was higher than 90%, and it could tolerate concentrations of chloride or sulfate up to 2%. The EtSeCN was extracted in chloroform and could be detected in electron ionization and also in negative chemical ionization mode with a further gain in signal-to-noise ratio by a factor of 2. The method was applied for the analysis of natural waters with quantitation of SeCN- in the low ng/g region. The Se13C15N- internal standard could be used for isotope dilution. Quantitative spike recoveries of 1 ng/g Se were obtained from seawater and river water, and 1 ng/g Se could be quantified within a standard uncertainty of 15%.

Selenium analysis in waters. Part 1: Regulations and standard methods.

Selenium is released into the aquatic environment through anthropogenic activities such as agricultural irrigation, coal mining, and metallurgical activities, where it acts as a reproductive toxin with negative effects on predatory fish and water fowl. Waterborne selenium concentrations are closely regulated worldwide, and various standardized methods are implemented by regulatory bodies to allow for the monitoring of selenium concentrations in different types of waters. Here, we discuss worldwide regulations relating to concentration limits of selenium in drinking, natural, and industrial waters. Focusing specifically on North America, we look at some standardized analysis methods and discuss the fact that many of these methods are not adequately sensitive to measure selenium in the concentrations outlined by the associated regulations for natural waters. We look in detail at the limitations of these methods with regards to both detection limits and interfering sample matrix components and establish the need for more sensitive and robust methods of analysis for regulatory compliance. This review is complemented by a second part (LeBlanc et al., 2018) where we discuss the state of selenium speciation analysis and importance of speciation data for decision makers in industry and regulators.

Selenium analysis in waters. Part 2: Speciation methods.

In aquatic ecosystems, there is often no correlation between the total concentration of selenium present in the water column and the toxic effects observed in that environment. This is due, in part, to the variation in the bioavailability of different selenium species to organisms at the base of the aquatic food chain. The first part of this review (Kumkrong et al., 2018) discusses regulatory framework and standard methodologies for selenium analysis in waters. In this second article, we are reviewing the state of speciation analysis and importance of speciation data for decision makers in industry and regulators. We look in detail at fractionation methods for speciation, including the popular selective sequential hydride generation. We examine advantages and limitations of these methods, in terms of achievable detection limits and interferences from other matrix species, as well as the potential to over- or under-estimate operationally-defined fractions based on the various conversion steps involved in fractionation processes. Additionally, we discuss methods of discrete speciation (through separation methods), their importance in analyzing individual selenium species, difficulties associated with their implementation, as well as ways to overcome these difficulties. We also provide a brief overview of biological treatment methods for the remediation of selenium-contaminated waters. We discuss the importance of selenium speciation in the application of these methods and their potential to actually increase the bioavailability of selenium despite decreasing its total waterborne concentration.

A computer program to simplify analysis of mass scan data of organometallic compounds from high-resolution mass spectrometers.

RATIONALE: Software accompanying high-resolution mass spectrometers, particularly that used for the analysis of organometallic compounds, has lagged the technology of the instruments themselves. We have developed a computer program that partially fills this gap. METHODS: Given the user's expectation for the number of atoms of a target element likely to be in an ion, the program calculates isotopologue mass differences for combinations of that element's isotopes and their expected intensity ratios relative to the most abundant isotopologue. These values are compared with mass differences and intensity ratios found in the experimental mass scan data and these metrics feed into a four-factor scoring model which ranks the ions as to the likelihood of each containing the specified number of the target atoms. The program was tested using experimental data obtained for selenomethionine. RESULTS: Across a broad range of sample concentrations, the program consistently ranked selenomethionine at or near the top of the list of ions that passed the screening and ranking process. Mass scan data files in excess of 24,000 records were analyzed in less than one second. CONCLUSIONS: The program is quick and efficient at scanning voluminous experimental data files for the presence of ions containing the expected number of atoms of a target element. Best results were obtained the scarcer the target element and the more isotopes it comprised. Copyright © 2016 John Wiley & Sons, Ltd.

Production and Release of Selenomethionine and Related Organic Selenium Species by Microorganisms in Natural and Industrial Waters.

Laboratory algal cultures exposed to selenate were shown to produce and release selenomethionine, selenomethionine oxide, and several other organic selenium metabolites. Released discrete organic selenium species accounted for 1.6-13.1% of the selenium remaining in the media after culture death, with 1.3-6.1% of the added selenate recovered as organic metabolites. Analysis of water from an industrially impacted river collected immediately after the death of massive annual algal blooms showed that no selenomethionine or selenomethionine oxide was present. However, other discrete organic selenium species, including a cyclic oxidation product of selenomethionine, were observed, indicating the previous presence of selenomethionine. Industrial biological treatment systems designed for remediation of selenium-contaminated waters were shown to increase both the concentration of organic selenium species in the effluent, relative to influent water, and the fraction of organic selenium to up to 8.7% of the total selenium in the effluent, from less than 1.1% in the influent. Production and emission of selenomethionine, selenomethionine oxide, and other discrete organic selenium species were observed. These findings are discussed in the context of potentially increased selenium bioavailability caused by microbial activity in aquatic environments and biological treatment systems, despite overall reductions in total selenium concentration.

Identification of trace levels of selenomethionine and related organic selenium species in high-ionic-strength waters.

A new anion-exchange chromatographic separation method was used for the simultaneous speciation analysis of selenoamino acids and the more ubiquitous inorganic selenium oxyanions, selenite and selenate. For quantification, this separation was coupled to inductively coupled plasma-mass spectrometry to achieve an instrumental detection limit of 5 ng Se L(-1) for all species. This chromatographic method was also coupled to electrospray tandem mass spectrometry to observe the negative ion mode fragmentation of selenomethionine and one of its oxidation products. Low detection limits were achieved, which were similar to those obtained using inductively coupled plasma-mass spectrometry. An extensive preconcentration and cleanup procedure using cation-exchange solid-phase extraction was developed for the identification and quantification of trace levels of selenomethionine in environmental samples. Preconcentration factors of up to five were observed for selenomethionine, which in addition to the removal of high concentrations of sulphate and chloride from industrial process waters, allowed for an unambiguous analysis that would have been impossible otherwise. Following these methods, selenomethionine was identified at an original concentration of 3.2 ng Se L(-1) in samples of effluent collected at a coal-fired power plant's biological remediation site. It is the first time that this species has been identified in the environment, outside of a biological entity. Additionally, oxidation products of selenomethionine were identified in river water and laboratory algal culture samples. High-resolution mass spectrometry was employed to postulate the chemical structures of these species.

Production and release of selenocyanate by different green freshwater algae in environmental and laboratory samples.

In a previous study, selenocyanate was tentatively identified as a biotransformation product when green algae were exposed to environmentally relevant concentrations of selenate. In this follow-up study, we confirm conclusively the presence of selenocyanate in Chlorella vulgaris culture medium by electrospray mass spectrometry, based on selenium's known isotopic pattern. We also demonstrate that the observed phenomenon extends to other green algae (Chlorella kesslerii and Scenedesmus obliquus) and at least one species of blue-green algae (Synechococcus leopoliensis). Further laboratory experiments show that selenocyanate production by algae is enhanced by addition of nitrate, which appears to serve as a source of cyanide produced in the algae. Ultimately, this biotransformation process was confirmed in field experiments where trace amounts of selenocyanate (0.215 ± 0.010 ppb) were observed in a eutrophic, selenium-impacted river with massive algal blooms, which consisted of filamentous green algae (Cladophora genus) and blue-green algae (Anabaena genus). Selenocyanate abundance was low despite elevated selenium concentrations, apparently due to suppression of selenate uptake by sulfate, and insufficient nitrogen concentrations. Finally, trace levels of several other unidentified selenium-containing compounds were observed in these river water samples; preliminary suggestions for their identities include thioselenate and small organic Se species.