Isotopic measurements of carbon dioxide: the role of measurement science and standards.

Isotopic measurements provide valuable information about the origin of greenhouse gases - as carbon dioxide levels increase, there is a corresponding shift towards lighter isotopic composition similar to that of fossil fuels. Detecting such isotopic shifts, however, requires extremely precise measurements, which must also be globally reproducible in order to make reliable policy decisions. This feature article outlines the collective search for the ideal standard for carbon isotope measurements since the 1950s. This tragicomedy of errors, if you wish, has strengthened the reliability of today's measurements and has taken us from fictional oceans, to toilet seat marbles, and complex mathematical conventions that separate data from reliable results.

Production and certification of BOTS-1: bovine muscle-certified reference material for incurred veterinary drug residues.

A freeze-dried bovine muscle-certified reference material (CRM), known as BOTS-1 (DOI: https://doi.org/10.4224/crm.2018.bots-1 ), containing incurred residues of commonly used veterinary drugs was produced and certified for the mass fraction of eight veterinary drug residues. Value assignment was carried out using liquid chromatography tandem mass spectrometry (LC-MS/MS) methods in conjunction with isotope dilution and standard addition approaches involving stable isotope internal standards. Data from the National Research Council of Canada (NRC), Canadian Food Inspection Agency (CFIA), United States Department of Agriculture (USDA), and the Federal Office of Consumer Protection and Food Safety in Germany (BVL) were used for value assignment. Results for two drug residues were also obtained through an international inter-laboratory comparison CCQM-K141/P178 organized under the auspices of the International Bureau of Weights and Measures (BIPM). Quantitative NMR (1H-qNMR) was used to characterize primary standards of all veterinary drugs certified. The certified mass fractions of the veterinary drug residues were 490 ± 100 µg/kg for chlorpromazine, 44 ± 4.4 µg/kg for ciprofloxacin, 3.3 ± 1.4 µg/kg for clenbuterol, 9.5 ± 0.8 µg/kg for dexamethasone, 57 ± 4.8 µg/kg for enrofloxacin, 3.0 ± 0.4 µg/kg for meloxicam, 12.4 ± 1.2 µg/kg for ractopamine, and 2290 ± 120 µg/kg for sulfadiazine with expanded uncertainties quoted (95% confidence) which include the effects due to between-bottle inhomogeneity, instability during long-term storage and transportation, and characterization.

Preparation of 18O-labelled azaspiracids for accurate quantitation using liquid chromatography-mass spectrometry.

Azaspiracids (AZAs) are a group of polyether marine algal toxins known to accumulate in shellfish, posing a risk to human health and the seafood industry. Analysis of AZAs is typically performed using LC-MS, which can suffer from matrix effects that significantly impact the accuracy of measurement results. While the use of isotopic internal standards is an effective approach to correct for these effects, isotopically labelled standards for AZAs are not currently available. In this study, 18O-labelled AZA1, AZA2, and AZA3 were prepared by reaction with H218O under acidic conditions, and the reaction kinetics and sites of incorporation were studied using LC-HRMS/MS aided by mathematical analysis of their isotope patterns. Analysis of the isotopic incorporation in AZA1 and AZA3 indicated the presence of four exchangeable oxygen atoms. Excessive isomerization occurred during preparation of 18O-labelled AZA2, suggesting a role for the 8-methyl group in the thermodynamic stability of AZAs. Neutralized mixtures of 18O-labelled AZA1 and AZA3 were found to maintain their isotopic and isomeric integrities when stored at -20 °C and were used to develop an isotope-dilution LC-MS method which was applied to reference materials of shellfish matrices containing AZAs, demonstrating high accuracy and excellent reproducibility. Preparation of isotopically labelled compounds using the isotopic exchange method, combined with the kinetic analysis, offers a feasible way to obtain isotopically labelled internal standards for a wide variety of biomolecules to support reliable quantitation.

Thermal stability of cannabinoids in dried cannabis: a kinetic study.

This study was undertaken to quantitatively explore the effect of temperature on the degradation of cannabinoids in dried cannabis flower. A total of 14 cannabinoids were monitored using liquid chromatography and tandem mass spectrometry in temperature environments from - 20 to + 40 ∘C lasting up to 1 year. We find that a network of first-order degradation reactions is well-suited to model the observed changes for all cannabinoids. While most studies focus on high-temperature effects on the cannabinoids, this study provides high-precision quantitative assessment of room temperature kinetics with applications to shelf-life predictions and age estimates of cannabis products.

Site-specific carbon isotope measurements of vanillin reference materials by nuclear magnetic resonance spectrometry.

Vanillin, one of the world's most popular flavor used in food and pharmaceutical industries, is extracted from vanilla beans or obtained (bio)-synthetically. The price of natural vanillin is considerably higher than that of its synthetic alternative which leads increasingly to counterfeit vanillin. Here, we describe the workflow of combining carbon isotope ratio combustion mass spectrometry with quantitative carbon nuclear magnetic resonance spectrometry (13C-qNMR) to obtain carbon isotope measurements traceable to the Vienna Peedee Belemnite (VPDB) with 0.7‰ combined standard uncertainty (or expanded uncertainty of 1.4‰ at 95% confidence level). We perform these measurements on qualified Bruker 400 MHz instruments to certify site-specific carbon isotope delta values in two vanillin materials, VANA-1 and VANB-1, believed to be the first intramolecular isotopic certified reference material (CRMs).

Characterization of vanillin carbon isotope delta reference materials.

Stable carbon isotope ratio measurements are used to investigate the provenance of vanillin. In this study, a variety of commercial vanillin samples and vanilla products were analyzed to provide a frame of reference for the variability of carbon isotope delta values in various vanillin samples, with the results ranging from -20.6 to -36.7‰ relative to the Vienna Peedee Belemnite (VPDB). We present information on the development of two synthetic vanillin reference materials, VANA-1 and VANB-1, prepared in 0.75 g units in glass vials, to be used for the calibration of carbon isotope delta measurements of vanillin and other easily combustible organic materials. Characterization of 40 vials each of VANA-1 and VANB-1 was performed by three laboratories over several measurement sequences. The certified carbon isotope delta values are -31.30 ± 0.06‰ (VANA-1) and -25.85 ± 0.05‰ (VANB-1). These uncertainties, for the 95% confidence level, include considerations for measurement uncertainty, coherence of the reference materials used for calibration, batch homogeneity, and stability during storage and transportation. The results are traceable to the VPDB through a set of nine reference materials (IAEA-CH-6, USGS65, IAEA-600, NBS22, USGS61, IAEA-603, IAEA-610, IAEA-611, and IAEA-612). For up to date certified values, users should refer to doi.org/10.4224/crm.2022.vana-1 and doi.org/10.4224/crm.2022.vanb-1.

Determination of the Isotopic Composition of Zirconium Using MC-ICPMS and a Regression Model for Mass Bias Correction.

Measurements of zirconium isotopes provide insights into the formation of planetary bodies, dating nuclear explosions, or maintenance of nuclear reactors. Although many comparative measurements for the isotopic composition of zirconium have been performed using mass spectrometry, there is a lack of zirconium isotopic reference materials and only a single calibrated measurement has been reported to date by total evaporation thermal ionization mass spectrometry (TE-TIMS). In this study, we report an independent measurement of full zirconium isotopic composition in a new zirconium isotopic reference material by multicollector inductively coupled plasma MS using the regression method and two independent certified isotopic reference materials, NIST SRM 984 Rb and NIST SRM 987 Sr. Our results agree well with the recent TE-TIMS measurements and suggest a revision of the standard atomic weight of zirconium.

Discontinuity in the Realization of the Vienna Peedee Belemnite Carbon Isotope Ratio Scale.

By convention, carbon isotope ratios are expressed relative to VPDB defined by the calcite standard NBS19 in the 1980s. [See T. Coplen, Pure Appl. Chem. 1994, 66, 273-276.] To improve the realization of the VPDB scale, a second fixed point (lithium carbonate, LSVEC) was introduced in 2006 [T. Coplen et al. Anal. Chem. 2006, 78, 2439-2441], which is now known to be isotopically unstable. [Assonov, S. Rapid Commun. Mass Spectrom., 2018, 32, 827-830.] With the high-quality reference materials made available in 2020, it is now possible to realize the VPDB scale with high confidence. [Assonov, S. et al. Rapid Commun. Mass Spectrom., 2020, 34, e8867; Assonov, S. Rapid Commun. Mass Spectrom. 2021, 35, e9014; Qi, H. et al. Rapid Commun. Mass Spectrosc. 2021, 35, e9006.] Here, we report the analysis of 25 reference materials using isotope ratio combustion mass spectrometry, show the discontinuity between the values measured against the new IAEA reference materials and the values currently assigned to these reference materials on the VPDB2006, and provide a link bringing these materials onto the new VPDB2020.

Determination of the Isotopic Composition of Gadolinium Using Multicollector Inductively Coupled Plasma Mass Spectrometry.

In this study, we report independent measurements of all stable isotope ratios of gadolinium. Our study employs multicollector inductively coupled plasma mass spectrometry (MC-ICPMS) with National Research Council Canada (NRC) HALF-1 isotopic hafnium standard as a primary calibrator and surveys four commercial gadolinium materials, including a NRC candidate isotopic reference material, GADS-1. The isotopic composition of gadolinium is determined using the regression model without reliance on conventional normalizing isotope ratios or mass-dependent isotope ratio correction models. In this work, all gadolinium isotope ratios were obtained from 160Gd/158Gd which, in turn, was measured from hafnium 178Hf/177Hf either directly or indirectly through 167Er/166Er. The latter approach was used for the final determination of gadolinium isotopic composition, as it provides smaller combined uncertainty. We report high-precision measurements of the isotopic composition of gadolinium, which support a revised standard atomic weight. Isotope amount ratios of R152/158 = 0.008 20(2)k=1, R154/158 = 0.087 98(12)k=1, R155/158 = 0.596 81(63)k=1, R156/158 = 0.825 08(57)k=1, R157/158 = 0.630 60(22)k=1, and R160/158 = 0.879 10(60)k=1, and the atomic weight of Ar(Gd) = 157.2502(6)k=1 were obtained for gadolinium in GADS-1.

Particle Size Distributions for Cellulose Nanocrystals Measured by Transmission Electron Microscopy: An Interlaboratory Comparison.

Particle size is a key parameter that must be measured to ensure reproducible production of cellulose nanocrystals (CNCs) and to achieve reliable performance metrics for specific CNC applications. Nevertheless, size measurements for CNCs are challenging due to their broad size distribution, irregular rod-shaped particles, and propensity to aggregate and agglomerate. We report an interlaboratory comparison (ILC) that tests transmission electron microscopy (TEM) protocols for image acquisition and analysis. Samples of CNCs were prepared on TEM grids in a single laboratory, and detailed data acquisition and analysis protocols were provided to participants. CNCs were imaged and the size of individual particles was analyzed in 10 participating laboratories that represent a cross section of academic, industrial, and government laboratories with varying levels of experience with imaging CNCs. The data for each laboratory were fit to a skew normal distribution that accommodates the variability in central location and distribution width and asymmetries for the various datasets. Consensus values were obtained by modeling the variation between laboratories using a skew normal distribution. This approach gave consensus distributions with values for mean, standard deviation, and shape factor of 95.8, 38.2, and 6.3 nm for length and 7.7, 2.2, and 2.9 nm for width, respectively. Comparison of the degree of overlap between distributions for individual laboratories indicates that differences in imaging resolution contribute to the variation in measured widths. We conclude that the selection of individual CNCs for analysis and the variability in CNC agglomeration and staining are the main factors that lead to variations in measured length and width between laboratories.

Determination of the isotopic composition of lutetium using MC-ICPMS.

In this study, we report the first independent measurements of lutetium isotopic composition using multi-collector ICPMS in four commercial lutetium materials, including a new NRC candidate lutetium isotopic reference material, LUIS-1. The regression model was used to correct for the instrumental isotopic fractionation (mass bias) using NIST SRM 989 isotopic rhenium as the primary calibrator. The regression model is based on a short 15-min measurement sessions at varying ICP plasma power. Isotope ratio of R176/175 = 0.026553(11)k = 1, corresponding isotopic abundances of x176 = 0.025866(11)k = 1, x175 = 0.974134(11)k = 1, and an atomic weight of Ar(Lu) = 174.966693(13)k = 1 were obtained for lutetium in LUIS-1. Uncertainty estimation was performed using a mixture of modeling approaches in accordance with the JCGM 100:2008 "Guide to the Expression of Uncertainty in Measurement" and its Supplement 1. The relative contribution of the rhenium primary standard to the combined uncertainty of 176Lu/175Lu isotope ratio was 15%.

CRM rapid response approach for the certification of arsenic species and toxic trace elements in baby cereal coarse rice flour certified reference material BARI-1.

With recently legislated maximum levels of inorganic arsenic (iAs) in white and brown rice in Canada, the regulatory bodies are evaluating the need for regulation of As levels in infant food products. Rice is a major part of infants' diet, and therefore, the presence of As in this staple food causes concerns. So far, the scientific community was lacking suitable certified reference material (CRM) which could be used to assess the accuracy of developed analytical methods for As speciation in infants' food products. As a result, we have developed BARI-1, a baby cereal coarse rice flour reference material which was certified for total arsenic (0.248 ± 0.018 mg kg-1), cadmium (0.0134 ± 0.0014 mg kg-1), mercury (0.0026 ± 0.0003 mg kg-1), lead (0.0064 ± 0.0016 mg kg-1), inorganic As (0.113 ± 0.016 mg kg-1) and dimethylarsinic acid (DMA) (0.115 ± 0.010 mg kg-1), and reference value for monomethylarsonic acid (MMA) (0.0045 ± 0.0008 mg kg-1) was reported. We also observed trace amounts of an unknown As compound, with chromatographic retention time close to DMA. Participating laboratories were allowed to use their in-house-validated extraction and/or digestion methods, and the detection of total metals was done by ICP-MS whereas HPLC-ICP-MS was used for As speciation. Despite the diversity in sample preparation and quantitation methods, reported values were in good agreement. For iAs measurement, the comparison between hydride generation ICP-MS and HPLC-ICP-MS found iAs overestimation with the former method, possibly due to interference from DMA. The certification was accomplished with a CRM rapid response approach in collaborative, focused effort completing the CRM development in few months instead of the typical multiyear project. This approach allowed to respond to measurement needs in a timely fashion. Graphical abstract.

High-Precision Measurements of the Isotopic Composition of Common Lead Using MC-ICPMS: Comparison of Calibration Strategies Based on Full Gravimetric Isotope Mixture and Regression Models.

The determination of isotope ratios of lead finds many important applications in earth and planetary sciences. In this study, we report the first independent and fully calibrated absolute isotope ratio measurements of a common lead since the seminal work by the NIST in the 1960s, which has provided the widely used standards SRM 981 and SRM 982. This work employs MC-ICPMS for the absolute isotope ratio measurements, which were calibrated using two independent methods: the full gravimetric isotope mixture method based on the use of all four near-pure lead isotopes (the first application of this primary method to lead) and the regression method with NIST SRM 997 thallium isotopic standard as the primary calibrator. Both calibration methods provide results consistent to a few parts in 104, which were used to characterize isotope ratios of lead in an NRC candidate reference material, high-purity common lead HIPB-1.

Three certified sugar reference materials for carbon isotope delta measurements.

RATIONALE: For isotope delta analysis, it is preferable to have at least two matrix-matched reference materials whose isotope delta values encompass those of the samples to be analyzed. The National Research Council Canada (NRC) has developed three sugar Certified Reference Materials (CRMs), BEET-1 (beet sugar), GALT-1 (galactose), and FRUT-1 (fructose), to be collectively used for carbon isotope delta measurements in sugars, and other organic materials. METHODS: All materials were homogenized and packaged in glass ampules. All three sugar materials were analyzed at the NRC using elemental analyzer/isotope ratio mass spectrometry (EA/IRMS). Six additional laboratories also provided EA/IRMS measurements. Data from all laboratories were re-normalized using three international secondary reference materials (IAEA-CH-6, USGS40, and USGS62) included as blind samples in the inter-laboratory comparison, thus providing added quality control and robustness to the study. RESULTS: Re-normalized carbon isotope delta values from each laboratory were combined using a random laboratory effects statistical model with accounting of the correlations between the laboratory results due to the use of the same reference materials for calibration. The consensus δ(13 C) values and combined standard uncertainties which include effects due to characterization, homogeneity, and stability for BEET-1, GALT-1, and FRUT-1 are -26.02(7) ‰, -21.41(6) ‰, and -10.98(5) ‰, respectively, on the VPDB scale. CONCLUSIONS: Three new δ(13 C) sugar CRMs (BEET-1, GALT-1, and FRUT-1) were developed and are available from NRC. These three CRMs can be utilized as a set for daily δ(13 C) scale normalization of sugar-based or other organic materials in order to produce reliable δ(13 C) measurements.

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.

Novel Microcystins from Planktothrix prolifica NIVA-CYA 544 Identified by LC-MS/MS, Functional Group Derivatization and 15N-labeling.

Microcystins are cyclic heptapeptides from cyanobacteria that are potent inhibitors of protein phosphatases and are toxic to animals and humans. At present, more than 250 microcystin variants are known, with variants reported for all seven peptide moieties. While d-glutamic acid (d-Glu) is highly-conserved at position-6 of microcystins, there has been only one report of a cyanobacterium (Anabaena) producing microcystins containing l-Glu at the variable 2- and 4-positions. Liquid chromatography-mass spectrometry analyses of extracts from Planktothrix prolifica NIVA-CYA 544 led to the tentative identification of two new Glu-containing microcystins, [d-Asp3]MC-ER (12) and [d-Asp3]MC-EE (13). Structure determination was aided by thiol derivatization of the Mdha7-moiety and esterification of the carboxylic acid groups, while 15N-labeling of the culture and isotopic profile analysis assisted the determination of the number of nitrogen atoms present and the elemental composition of molecular and product-ions. The major microcystin analog in the extracts was [d-Asp3]MC-RR (1). A microcystin with an unprecedented high-molecular-mass (2116 Da) was also detected and tentatively identified as a sulfide-linked conjugate of [d-Asp3]MC-RR (15) by LC-HRMS/MS and sulfide oxidation, together with its sulfoxide (16) produced via autoxidation. Low levels of [d-Asp3]MC-RW (14), [d-Asp3]MC-LR (4), [d-Asp3,Mser7]MC-RR (11), [d-Asp3]MC-RY (17), [d-Asp3]MC-RF (18), [d-Asp3]MC-RR-glutathione conjugate (19), and [d-Asp3]MC-RCit (20), the first reported microcystin containing citrulline, were also identified in the extract, and an oxidized derivative of [d-Asp3]MC-RR and the cysteine conjugate of 1 were partially characterized.

Determination of the Isotopic Composition of Osmium Using MC-ICPMS.

Despite its widespread applications in geology, all osmium isotope ratio measurements are either uncalibrated or rely on the veracity of the uncalibrated 1937 Nier values by adopting them as normalizing constants typically in conjunction with an exponential mass bias correction model. In this study, isotope ratios of osmium were determined in six commercial osmium materials, including the DROsS standard and a new NRC isotopic osmium reference material OSIS-1, by MC-ICPMS. We use a previously optimized and validated regression mass bias correction model to correct instrumental isotope fractionation effects which does not rely either on Nier's values or on a strictly mass-dependent behavior of the isotopes. Deviations from mass-dependent fractionation (mass independent fractionation) were observed for osmium isotopes in MC-ICPMS with the most dramatic effect occurring for 187Os, wherein, on average, close to half-percent bias in the isotope ratio 187Os/188Os was observed as a result of imposing Russell's law.