From sea salt to seawater: a novel approach for the production of water CRMs.

Natural water certified reference materials (CRMs) are mostly available in a liquid form, and they are produced starting from suitable environmental samples. Many precautions are usually needed to avoid biological or physical degradation, including filtration, acidification, and sterilization. In this study, the drawbacks associated with liquid-based seawater CRMs were tackled by developing a salt-based seawater proxy for nutrients that could be reconstituted in water solution just before use. Phosphate, silicate, and nitrate were chosen as target analytes. Sea salt mimicking the composition of seawater was spiked with an aqueous solution of the analytes and homogenized using a high-energy planetary ball mill (uhom < 1.2%). The salt powder CRM SALT-1 ( https://doi.org/10.4224/crm.2022.salt-1 ) demonstrated good short- and long-term stability for nutrients. When the SALT-1 was reconstituted in water at the 4.0% w/w level, the resulting solution had similar properties with respect to typical seawater in terms of major constituents (± 20%), trace metals, density (1.023 g/mL), pH (8.8-9.0), and optical properties relevant to the photometric characterization. Phosphate and silicate were quantified by photometry (molybdenum blue method, batch mode), whereas nitrate was quantified by isotope dilution GC-MS (uchar < 1.2%). In the SALT-1 reconstituted seawater solution at the 4.0% w/w salt level, the nutrient amount concentration was w(phosphate, PO43-) = 1.615 ± 0.030 µmol/L, w(silicate as SiO2) = 8.89 ± 0.31 µmol/L, and w(nitrate, NO3-) = 18.98 ± 0.45 µmol/L at the 95% confidence (k = 2). Overall, the SALT-1 CRM exhibits similar nutrient profile and general analytical characteristics as the MOOS-3 CRM. However, the SALT-1 has much reduced preparation, storage, and distribution cost, likely much better long-term stability, and it could enable the production of lower cost and more accessible seawater reference materials.

Application of direct analysis in real time to study chemical vapor generation mechanisms: reduction of dimethylarsinic(V) acid with aqueous NaBH4 under non-analytical conditions.

The aqueous-phase reaction of dimethylarsinc acid (DMAs(V)) with NaBH4 (THB) was studied under non-analytical conditions (1000 µg/mL As, 0.1 M HCl, 1% NaBH4) with the aim of identifying intermediates and reaction products. The use of direct analysis in real time (DART) with high-resolution mass spectrometry (HRMS), in combination with two different chemical vapor generation systems, allowed the identification of some species not detected by GC-MS such as Me2As-AsMe-AsMe2 and the arsonium species [Me3As-AsMe2]+ and [Me2As-AsMe2-AsMe2]+. Many other methylated species of arsenic containing up to four arsenic atoms have been observed. Unfortunately, the oxidation mechanism that took place in the DART source interfered with the identification of some of those species formed in solution following THB reduction. The species identified by DART-HRMS, together with those previously identified by GC-MS (Me2AsH, Me2AsOH, Me3As, Me3AsO, Me2AsAsMeH, Me2AsAsMe2, and Me2As-O-AsMe2)' enabled the formulation of hypotheses on the possible reaction pathways and revealed an aqueous-phase reactivity of DMAs(V) which could not be explained on the basis of current knowledge. Graphical Abstract.

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.

Versatile derivatization for GC-MS and LC-MS: alkylation with trialkyloxonium tetrafluoroborates for inorganic anions, chemical warfare agent degradation products, organic acids, and proteomic analysis.

Analytical chemists resort to derivatization for improving the detection performance of certain categories of analytes. Within this context, alkylation reactions are regarded as an important asset for many methods based on GC-MS and LC-MS. Trialkyloxonium tetrafluoroborates (R[Formula: see text][BF4]-) are powerful alkylating agents with ionic liquid properties: they are nonvolatile salts soluble in water which are easier and safer to handle with respect to common alkylating agents like diazomethane. R[Formula: see text][BF4]- can perform the alkylation in both organic and aqueous media at pH conditions ranging from acidic to alkaline. Recent analytical applications of trialkyloxonium derivatizations include the high-precision determination of inorganic anions in complex matrices, the qualitative confirmation of chemical warfare agent degradation products in soils, the profiling of carboxylic acids in urine, and the detection of protein post-translational modifications induced by carbon dioxide. The common denominator for all methods presented can be found in the simplicity of the alkylation protocol which, in most of the cases, requires a single step addition of the reagent directly to the sample. Graphical Abstract Alkylation with trialkyloxonium salts for GC-MS and LC-MS analysis.

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%.

Certification of nitrate in spinach powder reference material SPIN-1 by high-precision isotope dilution GC-MS.

A high-precision exact-matching quadruple isotope dilution method (ID4MS) was employed for the quantitation of nitrate in an air-dried spinach powder Certified Reference Material (CRM). The analyte was extracted in hot water following addition of 15NO[Formula: see text] internal standard. The blend was then treated with sulfamic acid to remove nitrite and with triethyloxonium tetrafluoroborate to promote aqueous conversion of nitrate into volatile EtONO2. The derivative was analyzed by headspace GC-MS with 3-min elution time. The method performance was validated with a series of tests which demonstrated adequate selectivity and ruggedness. This method supported the development of novel SPIN-1 CRM giving a modest contribution to its uncertainty (uchar = 0.85%). With respect to previous attempts, the SPIN-1 was proven stable, homogeneous (uhom = 0.44%), and suitable for spinach monitoring under EU regulations. On dried basis, the nitrate content of SPIN-1 was found to be 22.53 ± 0.43 mg/g (Uc = 1.9%, k = 2). The material was also used in an inter-laboratory study where four laboratories employed a total of ten measurement methods. Graphical Abstract SPIN-1 Certified Reference Material for nitrate in spinach powder.

A rapid and sensitive method for the determination of inorganic chloride in oil samples.

A novel isotope dilution method for the analysis of inorganic chloride in fuel oil matrix is presented. The samples were diluted by isopropyl alcohol:toluene, mixed with 37Cl- internal standard and reacted with triethyloxonium tetrafluoroborate at room temperature. This reagent promoted conversion of Cl- into stable ethyl chloride (EtCl) which was selectively detected by headspace GC-MS/MS with no matrix effects. A limit of detection of 0.2 mg kg-1 Cl- was obtained in fuel oil. The method was tested on the NIST SRM 1634c (trace elements in residual fuel oil) and validated through a series of robustness tests. Over 18 days, the variation in the signal response was less than 10% and the RSD for quantitative isotope dilution results was below 3%. Overall, the method is fast, simple and allows robust quantitation of inorganic chloride directly in the non-aqueous media.

Determination of elevated levels of nitrate in vegetable powders by high-precision isotope dilution GC-MS.

A high-precision isotope dilution GC-MS method was employed for the determination of nitrate in processed vegetables. The samples were extracted in water, derivatized with triethyloxonium tetrafluoroborate and analyzed by headspace GC-MS (15 samples/h). The method was applied to estimate the effect of drying on the content of nitrate in vegetables. The absolute amount of nitrate in a spinach sample before and after drying did not change even when the material was baked at 105 °C. Elevated levels of nitrate were found in commercial vegetable powders where the nitrate mass fraction exceed the percent level: 1.2-2.3% NO3- was found in spinach powders, 1.3-1.6% in kale powders, and 1.4% in a beetroot powder. The likely reduction of the antioxidant properties seen in vegetable powders along with their high nitrate content suggest the need to study the risk of endogenous N-nitrosation associated with these products.

Application of direct analysis in real time to the study of chemical vapor generation mechanisms: identification of intermediate hydrolysis products of amine-boranes.

In order to elucidate controversial results emerging in chemical vapor generation (CVG) for trace element determination, we conducted a series of experiments devoted to the identification of intermediates formed by acid hydrolysis of amine-boranes. For the first time, direct analysis in real time coupled with high-resolution mass spectrometry (DART-Orbitrap) was applied for detection of this class of compounds. Mass spectra of both solid amine-boranes and their aqueous solutions (pH ~ 8, no hydrolysis) were acquired for understanding their ionization pathway. Mass spectra of aqueous solutions of t-BuNH2·BH3 and Me2NH·BH3 were acquired under conditions that are employed in CVG (0.017-4.0 mol L-1 HCl, 0.167-0.2 mol L-1 borane reagent). The results disclose a reactivity driven by pH of amine-boranes undergoing hydrolysis. At low acidity, the hydrolysis proceeds according to the currently accepted displacement mechanisms (i.e., R3N·BH3 + H3O+ → R3NH+ + H2OBH3). At higher acidity, N-tert-butyl, cyclotriborazane, and bis(dimethylamino)boronium were identified, for the first time, during the hydrolysis of t-BuNH2·BH3 and Me2NH·BH3, respectively. Formation of these intermediates was ascribed to a hydrolysis pathway starting with the ionization of the amine-borane, (i.e., R3N·BH3 + H3O+ → [(H2O)R3NBH2] + + H2). The new evidence explains the anomalous behavior observed in CVG by amine-borane derivatization, and updates the currently accepted mechanisms for the acid hydrolysis of amine-boranes. Graphical Abstract.

Determination of total dissolved nitrogen in seawater by isotope dilution gas chromatography mass spectrometry following digestion with persulfate and derivatization with aqueous triethyloxonium.

In this study, we propose a novel approach for the determination of total dissolved nitrogen (TDN) in seawater combining high-precision isotope dilution GC-MS with persulfate digestion. A 2 mL sample aliquot was digested with an alkaline solution of persulfate to convert nitrogen containing compounds to nitrate. Digested samples were spiked with 15NO3- internal standard and treated with aqueous triethyloxonium to convert the analyte into volatile EtONO2. This derivative was readily separated from the matrix under gaseous form and could be sampled from the headspace before GC-MS analysis. The resulting chromatograms showed a stable flat baseline with EtONO2 as the only eluting peak (retention time 2.75 min on a DB 5.625 column). Such an approach provides specificity and obviates the shortcomings of current detection methods employed to analyze seawater samples after digestion with persulfate. In negative chemical ionization mode, the method reached a detection limit of 0.5 µmol/kg TDN (7 ng/g N) and could be applied to quantify seawater samples with 1-25 µmol/kg TDN. On the upper end of the range, quantitation could be repeated within 1%, whereas on a 6 µmol/kg TDN sample repeatability was 2.3% on eight measurements. The method was employed in two proficiency testing exercises providing results in agreement with consensus values. We investigated the impact of reagent blank and we implemented a blank-matching optimal design to account for such contribution. Finally, we performed a study on the yield of persulfate oxidation for organic and inorganic nitrogen compounds typically present in seawater. Whilst nitrite and ammonium are fully converted to nitrate, more complex organic molecules showed recoveries varying from 70% to 100%.

Determination of thiocyanate in exhaled breath condensate.

Thiocyanate is a heme peroxidase substrate that scavenges oxidants produced during inflammation and regulates host defense. In cystic fibrosis (CF) patients, increased airway thiocyanate levels are associated with improved lung function. Research on airway thiocyanate is limited, however, because convenient non-invasive airway sampling methods, such as exhaled breath condensate (EBC), yield low concentrations that are difficult to detect with available assays. In the present study, we developed a method for the determination of thiocyanate in dilute samples using isotope dilution headspace gas chromatography-coupled high-resolution, accurate-mass mass spectrometry (GC-HRMS). The method reliably quantified as little as 4 pmol thiocyanate in EBC and could detect even lower amounts. We successfully measured thiocyanate in EBC from seven healthy donors, with a mean ±â€¯SD of 27 ±â€¯16 nM and a median inter-assay coefficient of variation of 10.4% over six months. The method was applied to other biological fluids (plasma from the same visit as EBC donation; bronchoalveolar lavage fluid [BALF] from infants with CF; and healthy adult mouse BALF), giving reliable quantification of samples ranging from 10 nM to 100 µM. Thiocyanate concentrations in fluids besides EBC were (from lowest to highest): 0.73 ±â€¯0.39 µM in BALF of healthy adult mice (n = 6); 1.4 ±â€¯1.4 µM in BALF from infants with CF (n = 24); 46 ±â€¯22 µM in the plasma of adult volunteers (n = 7). These results demonstrate the utility of this new method for clinical determination of thiocyanate in EBC and other biological fluids.

Derivatization chemistries for the determination of inorganic anions and structurally related compounds by gas chromatography - A review.

Gas chromatography (GC) methods for the determination of inorganic anions and structurally related compounds are reviewed. In their native form, such analytes are polar and non-volatile, therefore they require derivatization before GC analysis. Several chemistries have been employed to convert anions to volatile molecules with applications to a wide set of analytes: nitrite, nitrate, halides, azide, bromate, iodate, borate, carbonate, thiocyanate, cyanide, sulfide, silicates, phosphates, phosphonates, selenite, selenate, arsenite, arsenate, monomethylarsonic acid, and dimethylarsinic acid have been measured following GC separation. In this review, most derivatization chemistries employed for anions are discussed with attention to molecular aspects of the conversion, experimental conditions, applications to complex sample matrices, and figure of merits.

Inter-laboratory study for the certification of trace elements in seawater certified reference materials NASS-7 and CASS-6.

Certification of trace metals in seawater certified reference materials (CRMs) NASS-7 and CASS-6 is described. At the National Research Council Canada (NRC), column separation was performed to remove the seawater matrix prior to the determination of Cd, Cr, Cu, Fe, Pb, Mn, Mo, Ni, U, V, and Zn, whereas As was directly measured in 10-fold diluted seawater samples, and B was directly measured in 200-fold diluted seawater samples. High-resolution inductively coupled plasma mass spectrometry (HR-ICPMS) was used for elemental analyses, with double isotope dilution for the accurate determination of B, Cd, Cr, Cu, Fe, Pb, Mo, Ni, U, and Zn in seawater NASS-7 and CASS-6, and standard addition calibration for As, Co, Mn, and V. In addition, all analytes were measured using standard addition calibration with triple quadrupole (QQQ)-ICPMS to provide a second set of data at NRC. Expert laboratories worldwide were invited to contribute data to the certification of trace metals in NASS-7 and CASS-6. Various analytical methods were employed by participants including column separation, co-precipitation, and simple dilution coupled to ICPMS detection or flow injection analysis coupled to chemiluminescence detection, with use of double isotope dilution calibration, matrix matching external calibration, and standard addition calibration. Results presented in this study show that majority of laboratories have demonstrated their measurement capabilities for the accurate determination of trace metals in seawater. As a result of this comparison, certified/reference values and associated uncertainties were assigned for 14 elements in seawater CRMs NASS-7 and CASS-6, suitable for the validation of methods used for seawater analysis.

Determination of cyanocobalamin by isotope dilution LC-MS/MS.

Cyanocobalamin (CNCbl) is an active form of vitamin B12, commonly employed for the preparation of multivitamin supplements and fortified food. In this study, we present a novel analytical method for its determination based on stable isotope dilution liquid chromatography electrospray tandem mass spectrometry (ID LC-MS/MS). Isotopically enriched 13C15NCbl was synthesized in-house and used as internal standard. The method was validated using NIST SRM 3280 multivitamin reference material and by comparison with an independent methodology based on LC-ICPMS. The proposed method provided a detection limit of 57 pg/g and could be applied for the determination of trace level of CNCbl in multivitamin supplements with a relative standard uncertainty better than 3%. The novel ID LC-MS/MS is a primary ratio method that could become a reference for CNCbl determination in multivitamins and food supplements. The method was applied for the characterization of two NRC multivitamin tablet Certified Reference Material (CRM) candidates, VITA-1 and VITB-1 whose CNCbl levels were quantified as 2.64 ± 0.09 and 1.75 ± 0.12 µg/g, respectively.

Rapid determination of nitrate in vegetables by gas chromatography mass spectrometry.

In this study we present a novel isotope dilution gas chromatography method for the determination of nitrate in vegetables. The analyte was extracted in water at 70∘C and mixed with 15N isotopically enriched nitrate internal standard. The sample was centrifuged and the supernatant reacted with sulfamic acid for removal of nitrite, and with triethyloxonium tetrafluoroborate for converting nitrate into volatile EtONO2. This simple aqueous chemistry allowed for separation of analyte from sample matrix in the form of a gaseous derivative which could be sampled in the headspace before GC-MS analysis. This key-feature of the method made possible the collection of clean chromatograms within an elution time of only 1.8 min. Detection of EtONO2 could be performed using electron impact ionization with a standard GC-MS setup. The method was optimized and validated for the analysis of nitrate in fresh vegetables in the 10-10,000 µg/g range with a detection limit of 2 µg/g. Due to the use of primary isotope dilution quantitation, traceable results of high-precision were attained.

Determination of total cyanide in soil by isotope dilution GC/MS following pentafluorobenzyl derivatization.

The high toxicity of cyanide, along with its widespread industrial use, has fuelled interest in the development of analytical methods for its determination in complex matrices. In this study, we propose a novel approach for the measurement of total cyanide in soil samples based on single-step derivatization with pentafluorobenzyl bromide (F5Bn-Br) followed by quantitation with gas chromatography mass spectrometry in negative chemical ionization mode. The reaction between CN- and F5Bn-Br resulted in the identification of several derivatives such as F5Bn-CN, (F5Bn)(F5Ph)CH-CN, and (F5Bn)2(F5Ph)C-CN. The relative proportion between such compounds was dependent on experimental conditions. When a 100 µL aliquot of an alkaline-aqueous extract was reacted with 700 µL of 1.3% F5Bn-Br in acetone, the tri-alkylated derivative was the most abundant. In such conditions a detection limit of 0.5 ng/g of CN- was attained. Soil samples were initially spiked with an alkaline solution of K13C15N internal standard and suspended in 7.5% aqueous NaOH. Determination of total cyanide was achieved by digestion of the alkaline extract with H3PO4 to produce HCN which was then trapped in 0.1% NaOH in a sealed double vial system, followed by reaction with F5Bn-Br. Isotope dilution calibration was chosen for quantitation, and the validity of the novel method was demonstrated by analysis of soil Certified Reference Materials (CRMs) and by spike recovery tests.