Isotope Ratio Mass Spectrometry Research Articles

Development of a Matrix‐Matched Barite Reference Material (NWU‐Brt) for Calibration of In Situ S Isotope Measurements by Laser Ablation Multi‐Collector Inductively Coupled Plasma‐Mass Spectrometry

A new matrix‐matched reference material (NWU‐Brt) with sulfur isotope ratios resembling those of natural barites has been developed for in situ S isotope measurements by laser ablation multi‐collector inductively coupled plasma‐mass spectrometry (LA‐MC‐ICP‐MS). A 100 g quantity of natural barite crystal was milled to ultra‐fine particles and sintered to a solid block using a fast hot‐pressing sintering technique (FHPS). We report δ34S ratios determined by isotope ratio mass spectrometry (IRMS), solution nebuliser multi‐collector inductively coupled plasma‐mass spectrometry (SN‐MC‐ICP‐MS) and LA‐MC‐ICP‐MS, involving up to six participating laboratories. The homogeneity of δ34S ratios of the synthesised barite was tested by LA‐MC‐ICP‐MS with an analytical spot size of 53 μm. The LA‐MC‐ICP‐MS results show that NWU‐Brt demonstrates a satisfactory homogeneous composition and is an appropriate material for calibrating δ34S ratios in barite. IRMS and SN‐MC‐ICP‐MS produced mean δ34S values of +14.17 ± 0.42‰ (2s) and +14.27 ± 0.23‰ (2s), respectively. The results of LA‐MC‐ICP‐MS analyses are consistent with the IRMS and SN‐MC‐ICP‐MS ratios within uncertainty. Overall, our results confirm the suitability of NWU‐Brt for the calibration of δ34S ratios in barite using LA‐MC‐ICP‐MS.

Affordable event and monthly rain samplers: Improving isotopic datasets to understand meteorological processes.

Water-stable isotopes in rainfall are powerful tracers of atmospheric processes at different spatial and temporal scales. However, commercially available rain samplers for isotopic analysis are prohibitively expensive, especially for high spatial resolution networks and studies conducted in developing countries. A low-cost, simple, and robust sampler was designed for event and monthly rainfall samplings. Rainfall collectors were built based on existing designs provided in the literature and using easily accessible materials. Event samplers were filled with different volumes of reference water and left for 72 h in laboratory conditions to determine the minimum amount of rainfall to be collected to minimize isotopic fractionation, from both postsampling evaporation and equilibration. Samples were analyzed using dual-inlet isotope ratio mass spectrometry and cavity ring-down spectroscopy. For samples larger than 4% of the bottle's capacity, the evaporative enrichment due to Rayleigh distillation is negligible compared to the overall analytical uncertainty. Using a tube connecting the funnel to the water sample has proved to reduce postsampling evaporation by at least five times. To limit water self-diffusion, we recommend collecting the largest rainfall amount possible. Under these conditions, these collectors are suitable for rainfall sampling for isotopic analysis. This low-cost methodology will enable isotopic sampling of precipitation at high spatial resolutions and democratize the use of isotopes for hydrological studies in developing countries. All instructions for building and using these samplers are made openly accessible to the scientific community so they can be repeated and adapted to the needs of each project.

Prevalence of nandrolone preparations with endogenous carbon isotope ratios in Australia

AbstractNandrolone and its prohormones, including 19‐norandrost‐4‐ene‐3,17‐dione and 19‐norandrost‐4‐ene‐3β,17β‐diol, are anabolic steroids forbidden at all times in sports according to the World Anti‐Doping Code Prohibited List and its metabolite 19‐norandrosterone (19NA) is the preferred urinary target compound to identify their abuse. In recent years, an increasing number of 19NA isotope ratio mass spectrometry (IRMS) cases have arisen that, based on the initial testing procedure, were likely to result in an adverse analytical finding but were concluded negative after IRMS analysis. The current study was therefore set up to gain a better insight on the prevalence of nandrolone preparations with endogenous carbon isotope ratio values in Australia. Suitable workplace (non‐athlete) urine samples that had previously been reported positive for 19NA were identified and analysed on IRMS. A total of 82% of the samples that were analysed were reported with enriched carbon isotope ratios of 19NA (i.e., 19NA greater than −26‰). This indicates that there is a high prevalence of nandrolone‐containing anabolic androgenic steroid preparations in Australia that have ‘endogenous’ carbon isotope ratios which reduces the ability to identify exogenous nandrolone.

Influence of microbially fermented 2´-fucosyllactose on neuronal-like cell activity in an in vitro co-culture system.

2´-Fucosyllactose (2´-FL), the most abundant oligosaccharide in human milk, plays an important role in numerous biological functions, including improved learning. It is not clear, however, whether 2´-FL or a cleavage product could influence neuronal cell activity. Thus, we investigated the effects of 2´-FL, its monosaccharide fucose (Fuc), and microbial fermented 2´-FL and Fuc on the parameters of neuronal cell activity in an intestinal-neuronal transwell co-culture system in vitro. Native 13C-labeled 2´-FL and 13C-Fuc or their metabolites, fermented with Bifidobacterium (B.) longum ssp. infantis and B. breve, which were taken from the lag-, log- and stationary (stat-) growth phases of batch cultures, were applied to the apical compartment of the co-culture system with Caco-2 cells representing the intestinal layer and all-trans-retinoic acid-differentiated SH-SY5Y (SH-SY5YATRA) cells mimicking neuronal-like cells. After 3 h of incubation, the culture medium in the basal compartment was monitored for 13C enrichment by using elemental analysis isotope-ratio mass spectrometry (EA-IRMS) and effects on cell viability, plasma, and mitochondrial membrane potential. The neurotransmitter activation (BDNF, GABA, choline, and glutamate) of SH-SY5YATRA cells was also determined. Furthermore, these effects were also measured by the direct application of 13C-2´-FL and 13C-Fuc to SH-SY5YATRA cells. While no effects on neuronal-like cell activities were observed after intact 2´-FL or Fuc was incubated with SH-SY5YATRA cells, supernatants from the stat-growth phase of 2´-FL, fermented by B. longum ssp. infantis alone and together with B. breve, significantly induced BDNF release from SH-SY5YATRA cells. No such effects were found for 2´-FL, Fuc, or their fermentation products from B. breve. The BDNF release occurred from an enhanced vesicular release, which was confirmed by the use of the Ca2+-channel blocker verapamil. Concomitant with this event, 13C enrichment was also observed in the basal compartment when supernatants from the stat-growth phase of fermentation by B. longum ssp. infantis alone or together with B. breve were used. The results obtained in this study suggest that microbial products of 2´-FL rather than the oligosaccharide itself may influence neuronal cell activities.

True Ileal Amino Acid Digestibility and Protein Quality of 15N-Labeled Faba Bean in Healthy Humans

BackgroundThe recommended transition toward more plant-based diets, particularly containing legumes, requires a wider knowledge of plant protein bioavailability. Faba beans are cultivated at different latitudes and are used increasingly in human nutrition. ObjectivesWe aimed to assess the nutritional quality of faba bean protein in healthy volunteers equipped with an intestinal tube to implement the ileal 15N balance method. MethodsNine volunteers completed the study (7 males, 2 females, aged 33 ± 10 y, BMI: 24.7 ± 2.6 kg/m2). They were equipped with a nasoileal tube. After fasting overnight, they ingested a test meal consisting of cooked mash of dehulled faba bean seeds (20 g protein per serving of approximately 250 g) intrinsically labeled with 15N. Samples of ileal contents, plasma, and urine were collected over an 8-h postprandial period. Undigested nitrogen (N) and amino acids (AAs) were determined using isotopic MS, and subsequently, ileal digestibility and digestible indispensable amino acid score (DIAAS) were calculated. The measurement of postprandial deamination allowed calculation of the net postprandial protein utilization (NPPU). ResultsThe ileal N digestibility was 84.1% ± 7.7%. Postprandial deamination represented 19.2% ± 3.6% of ingested N, and the NPPU was 64.7% ± 9.7%. The ileal digestibility of individual AAs varied from 85.1% ± 13.7% for histidine to 94.2% ± 3.6% for glutamine + glutamate. The mean AA digestibility was ∼6 percentage points higher than the digestibility of N, reaching 89.8% ± 5.9%, whereas indispensable AA digestibility was 88.0% ± 7.3%. Histidine and tryptophan were the first limiting AAs [DIAAS = 0.77 (calculated by legume-specific N-to-protein conversion factor 5.4); 0.67 (by default factor 6.25)]. Sulfur AAs were limiting to a lesser extent [DIAA ratio = 0.94 (N × 5.4); 0.81 (N × 6.25)]. ConclusionsProtein ileal digestibility of cooked, dehulled faba beans in humans was moderate (<85%), but that of AAs was close to 90%. Overall protein quality was restricted by the limited histidine and tryptophan content.This trial was registered at clinicaltrials.gov as NCT05047757.

Identification of provenance of Basmati rice grown in different regions of Punjab through multivariate analysis

High-priced Basmati rice is vulnerable to deliberate mislabeling to increase profits. This type of fraud may lower consumers’ confidence as inferior products can affect brand reputation. To address this problem, there is a need to devise a method that can efficiently distinguish Basmati rice grown in regions that are famous versus the regions that are not suitable for their production. Therefore, in this investigation, thirty-six samples of Basmati rice were collected from two zones of Punjab province (one known for Basmati rice) of Pakistan which is the major producer of Basmati rice. The elemental composition of rice samples was assessed using inductively coupled plasma-optical emission spectrometry and an organic elemental analyzer, whereas data on δ13C was acquired using isotopic ratio-mass spectrometry. Regional clustering of samples based on their respective cultivation zones was observed using multivariate data analysis techniques. Partial least squares-discriminant analysis was found to be effective in grouping rice samples from the different locations and identifying unknown samples belonging to these two regions. Further recommendations are presented to develop a better model for tracing the origin of unidentified rice samples.

Naturally occurring radioactive materials in offshore infrastructure: Understanding formation and characteristics of baryte scale during decommissioning planning

Contaminants, including naturally occurring radioactive material (NORM) of the 238-uranium and 232-thorium decay series, have been recognized as a global research priority to inform offshore petroleum infrastructure decommissioning decisions. This study aimed to characterize pipeline scale retrieved from a decommissioned subsea well tubular pipe through high-resolution elemental mapping and isotopic analysis. This was achieved by utilizing transmission electron microscopy, Synchrotron x-ray fluorescence, photostimulated luminescence autoradiography and Isotope Ratio Mass Spectrometry. The scale was identified as baryte (BaSO4) forming a dense crystalline matrix, with heterogenous texture and elongated crystals. The changing chemical and physical microenvironment within the pipe influenced the gradual growth rate of baryte over the production life of this infrastructure. A distinct compositional banding of baryte and celestine (SrSO4) bands was observed. Radioactivity attributed by the presence of radionuclides (226Ra, 228Ra) throughout the scale was strongly correlated with baryte. From the detailed scale characterization, we can infer the baryte scale gradually formed within the internals of the tubular well pipe along the duration of production (i.e., 17 years). This new knowledge and insight into the characteristics and formation of petroleum waste products will assist with decommissioning planning to mitigate potential radiological risks to marine ecosystems.

Exploring δ13 C data compatibility in a broad δ13 C range: Reassembling δ13 C values obtained for NIST CO2 RMs 8562-8564 at different laboratories.

Stable isotope datasets obtained in different laboratories over years must be compared and combined. The strict compatibility in δ13 C at ±0.01‰ and ±0.02‰, k= 1, respectively, is required for observations of greenhouse gases CO2 and methane. That implies the long-term compatibility of calibrations realized at different laboratories. Thus far, this level of compatibility has not been demonstrated for pure CO2 in a broad δ13 C range and over years. We reassemble the δ13 C obtained in different laboratories for NIST CO2 RMs 8562-8564, which are pure CO2 gases. If the measurements are based on independent calibrations and reported on the same 17 O correction framework, δ13 C of all three RMs can be used to estimate the long-term δ13 C data compatibility. δ13 C values obtained in different laboratories for NIST CO2 RMs 8562-8564 (a range of δ13 C from ~ -3.7‰ to ~ -41.6‰) from 1998 to 2020 agree within their uncertainties. Two NIST studies (1999 and 2004) have combined data from several laboratories using different Isotope ratio mass spectrometry (IRMS) methods and somewhat different approaches. Three single-laboratory studies (2003, 2008, and 2020) have used the same type of IRMS and similar approaches. The δ13 C values from these laboratories agree within ±0.01‰, k= 1, over the entire δ13 C range and have uncertainties about two times lower than those obtained at NIST by combining data from several laboratories. The δ13 C values for NIST CO2 RMs 8562-8564 obtained in three laboratories using the same IRMS (2003, 2008, and 2020) agree within ±0.01‰, k= 1. First, this confirms the long-term stability in δ13 C of RMs 8562-8564. Second, this example of compatibility in δ13 C calibrations on pure CO2 , from -3.7‰ to -41.6‰, is relevant to stable isotope measurements of greenhouse gases CO2 and methane. Third, this confirms the high accuracy of the δ13 C(air-CO2 ) calibrations performed in 2003 and 2008 for the project Civil Aircraft for the Regular Investigation of the atmosphere Based on an Instrumented Container (CARIBIC).

Comparison of analytical approaches for the detection of oral testosterone undecanoate administration in men.

For antidoping laboratories, the determination of an illicit testosterone (T) administration in urine samples remains a difficult process as it requires the determination of the exogenous origin by carbon isotope ratios (CIRs) of testosterone and its metabolites. As a complement to the urinary analysis, targeting testosterone esters (e.g. testosterone undecanoate [TU]) in serum samples by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) could represent a simpler approach compared with isotope ratio mass spectrometry (IRMS). These two approaches both lead to the direct detection of the administration of exogenous T but with a difference in effort and complexity of the analysis. To compare the detection window obtained with the two strategies, serum and the corresponding urine samples collected from an administration study with oral TU were analysed. Results showed that, at all timepoints where the intact TU was detected in serum, the CIRs of urinary steroids were also not in agreement with an endogenous origin. IRMS analysis required more effort but resulted in slightly longer detection windows than the ester analysis. Finally, this comparison study showed that, in the presence of a suspicious urinary steroid profile, the LC-MS/MS steroid esters analysis in the corresponding serum samples can be very helpful. If steroid esters are not detected, the IRMS analysis can then be conducted on the urine sample afterwards. Overall, the combination of matrices might facilitate the detection of prohibited T administration in sports, especially for athletes with naturally low T/E ratios.

High-Temporal-Resolution In Situ Sensor for Oceanic CO2 Isotope Measurement Enabling Multidimensional Isotope Tracing Analysis (R13C, R18O, and R17O) via Laser Absorption Spectroscopy.

Combined in situ analysis of oceanic CO2 concentrations and diverse C and O isotope characteristics can offer a unique perspective with multiple isotopic tracing dimensions for identifying marine biogeochemical processes. Applying this strategy in marine environments is urgently required, yet it faces inherent challenges in terms of existing analytical methods and instruments, e.g., a lack of in situ sensors, limited detectable isotope variety, and low-temporal-resolution data. Here, we report an underwater in situ dissolved CO2 isotope sensor based on mid-infrared tunable diode laser absorption spectroscopy (MIR-TDLAS) and membrane extraction technology. Through the proposed targeted strategies, the sensor is capable of providing high-temporal-resolution in situ measurement of all monosubstituted isotopes of dissolved CO2 (16O13C16O, 18O12C16O, and 17O12C16O) at marine background concentrations. The sensor is demonstrated to provide comparable precision to that of isotope ratio mass spectrometry. At 400 ppmv, the precision for R13C, R18O, and R17O could achieve 0.084, 0.042, and 0.013‰, respectively, for a 1 s integration time. By enabling a high-frequency in situ analysis in fixed-point time-series field deployment, a 17O anomaly with strong regularity is observed, which is not obvious in 18O and 13C, and therefore, the superiority of the proposed multidimensional in situ isotope tracing strategy is demonstrated. The developed sensor has great potential to open up new prospects for advancing marine carbon research.

Application of an absolute reference frame for methane clumped-isotope analyses

Clumped-isotope analysis provides new constraints on the formation and evolution of methane. High-resolution isotope-ratio mass spectrometry (HR-IRMS) is a sensitive, precise, and stable analytical technique increasingly used in methane clumped-isotope analysis. However, the lack of standard reference frames hinders the comparison of data from different laboratories and the evaluation of accuracy. In this study, an absolute reference frame was introduced for HR-IRMS analysis of the multiply substituted methane isotopologues 13CH3D and 12CH2D2. We evaluated the optimized performance of a Panorama (Nu Instruments, UK) HR-IRMS system by repeated analysis of an intra-laboratory standard gas, finding that these rare isotopologues can be analyzed separately with high precision, repeatability, and reproducibility. Internal precision (a key parameter in instrument performance) reached 0.15‰ − 0.25‰ for Δ13CH3D and 0.70‰ − 0.85‰ for Δ12CH2D2 values, ensuring the reliability of data. Comparison of results before and after purification demonstrated that purification procedures caused negligible isotopic fractionation of clumped isotopes. Isotopic-exchange experiments revealed source effects in the MS ion source, depending on the difference in bulk H isotopic composition between sample and reference gas, but independent of bulk C isotopic composition. To improve analytical accuracy for methane isotopologues, an absolute reference frame was developed through experiments involving a suite of mixed gases of different H isotopic composition equilibrated at 100 °C and a standard gas equilibrated at 2 °C–500 °C. Based on the reference frame, raw data were processed with a non-linearity correction and converted to real values with empirical transformation functions. The reference frame was applied to analyses of two methane standard gases and three natural gases from Tarim Basin, China. Measured Δ13CH3D and Δ12CH2D2 values for two standard gases were consistent with published values. Clumped-isotope signatures of natural gases preserve information related to the formation temperature of methane.

Estimating the heritability of nitrogen and carbon isotopes in the tail hair of beef cattle

BackgroundThe natural abundance of nitrogen (δ15N) and carbon (δ13C) isotopes in animal tissues are used to estimate an animal’s efficiency in nitrogen utilization, and their feed conversion efficiency, especially in tropical grazing systems with prolonged protein restriction. It is postulated that selection for improving these two characteristics (δ15N and δ13C) would assist the optimisation of the adaptation in ever-changing environments, particularly in response to climate change. The aim of this study was to determine the heritability of δ15N and δ13C in the tail hair of tropically adapted beef cattle to validate their inclusion in genetic breeding programs.MethodsIn total, 492 steers from two breeds, Brahman (n = 268) and Droughtmaster (n = 224) were used in this study. These steers were managed in two mixed breed contemporary groups across two years (year of weaning): steers weaned in 2019 (n = 250) and 2020 (n = 242). Samples of tail switch hair representing hair segments grown during the dry season were collected and analysed for δ15N and δ13C with isotope-ratio mass spectrometry. Heritability and variance components were estimated in a univariate multibreed (and single breed) animal model in WOMBAT and ASReml using three generations of full pedigree.ResultsThe estimated heritability of both traits was significantly different from 0, i.e. 0.43 ± 0.14 and 0.41 ± 0.15 for δ15N and δ13C, respectively. These traits had favourable moderate to high genetic and phenotypic correlations (− 0.78 ± 0.16 and − 0.40 ± 0.04, respectively). The study also provides informative single-breed results in spite of the limited sample size, with estimated heritability values of 0.37 ± 0.19 and 0.19 ± 0.17 for δ15N and δ13C in Brahman, and 0.36 ± 0.21 and 0.46 ± 0.22 for δ15N and δ13C in Droughtmaster, respectively.ConclusionsThe findings of this study show, for the first time, that the natural abundances of both nitrogen and carbon isotopes in the tail hair in cattle may be moderately heritable. With further research and validation, tail hair isotopes can become a practical tool for the large-scale selection of more efficient cattle.

Mass spectrometry imaging of Arabidopsis thaliana with in vivo D2O labeling.

The commonly used analytical tools for metabolomics cannot directly probe metabolic activities or distinguish metabolite differences between cells and suborgans in multicellular organisms. These issues can be addressed by in-vivo isotope labeling and mass spectrometry imaging (MSI), respectively, but the combination of the two, a newly emerging technology we call MSIi, has been rarely applied to plant systems. In this study, we explored MSIi of Arabidopsis thaliana with D2O labeling to study and visualize D-labeling in three classes of lipids: arabidopsides, chloroplast lipids, and epicuticular wax. Similar to other stress responses, D2O-induced stress increased arabidopsides in an hour, but it was relatively minor for matured plants and reverted to the normal level in a few hours. The D-labeling isotopologue patterns of arabidopsides matched with those of galactolipid precursors, supporting the currently accepted biosynthesis mechanism. Matrix-assisted laser desorption/ionization (MALDI)-MSI was used to visualize the spatiotemporal distribution of deuterated chloroplast lipids, pheophytin a, MGDGs, and DGDGs, after growing day-after-sowing (DAS) 28 plants in D2O condition for 3-12 days. There was a gradual change of deuteration amount along the leaf tissues and with a longer labeling time, which was attributed to slow respiration leading to low D2O concentration in the tissues. Finally, deuterium incorporation in epicuticular wax was visualized on the surfaces of the stem and flower. The conversion efficiency of newly synthesized C30 aldehyde to C29 ketone was very low in the lower stem but very high at the top of the stem near the flower or on the flower carpel. This study successfully demonstrated that MSIi can unveil spatiotemporal metabolic activities in various tissues of A. thaliana.

Identification of Proteomic Biomarkers of Acetaminophen-Induced Hepatotoxicity Using Stable Isotope Labeling.

Quantitative proteomics approaches based on stable isotopic labeling and mass spectrometry have been widely applied to disease research, drug target discovery, biomarker identification, and systems biology. One of the notable stable isotopic labeling approaches is trypsin-catalyzed 18O/16O labeling, which has its own advantages of low sample consumption, simple labeling procedure, cost-effectiveness, and absence of chemical reactions that potentially generate by-products. In this chapter, a protocol for 18O/16O labeling-based quantitative proteomics approach is described with an application to the identification of proteomic biomarkers of acetaminophen (APAP)-induced hepatotoxicity in rats. The protocol involves first the extraction of proteins from liver tissues of control and APAP-treated rats and digestion into peptides by trypsin. After cleaning of the peptides by solid-phase extraction, equal amounts of peptides from the APAP treatment and the control groups are then subject to trypsin-catalyzed 18O/16O labeling. The labeled peptides are combined and fractionated by off-line strong cation exchange liquid chromatography (SCXLC), and each fraction is then analyzed by nanoflow reversed-phase LC coupled online with tandem mass spectrometry (RPLC-MS/MS) for identification and quantification of differential protein expression between APAP-treated rats and controls. The protocol is applicable to quantitative proteomic analysis for a variety of biological samples.

Elemental (CNS) and isotopic (δ13C and δ15N) characterization of local sources to atmospheric dust at Trombay, Mumbai, using elemental analyzer–isotope ratio mass spectrometry

Particulate matter (PM) in the air has significant implications for human health and the environment, particularly in urban areas with high concentrations. This study focuses on the isotopic (δ13C and δ15N), elemental (CNS) as well as the total carbon to total nitrogen ratio (TC/TN) characterization of PM in Trombay, Mumbai. Monthly atmospheric dust samples were collected from 2016 to 2020 during the premonsoon and postmonsoon seasons. The average elemental concentrations for premonsoon samples were N (1.73% ± 0.25%), C (7.38% ± 0.72%), S (0.84% ± 0.13%), and TC/TN ratio (4.38 ± 0.86). For postmonsoon samples, the average elemental concentrations were N (1.47% ± 0.185%), C (6.95% ± 1.04%), S (0.84% ± 0.13%), and TC/TN ratio (4.80 ± 0.94). The average δ13C values for premonsoon and postmonsoon seasons were −24.31 ± 1.15‰ and −24.56 ± 1.20‰, respectively. The average δ15N values for premonsoon and postmonsoon seasons were 5.22 ± 1.92‰ and 4.57 ± 1.32‰, respectively. The results showed variations in elemental concentrations between the seasons, with higher values observed in premonsoon samples which can be attributed to increased dust generation from land and sea. Overall, this study provides insights into the local sources and characteristics of PM in the Trombay area and highlights the importance of long-term monitoring to understand the seasonal variations in PM composition.

Two-dimensional high performance liquid chromatography purification of underivatized urinary prednisone and prednisolone for compound-specific stable carbon isotope analysis.

The gas chromatography-combustion isotope ratio mass spectrometry (GC/C/IRMS) confirmation procedure for prednisone (PS) and prednisolone (PSL) is still a great challenge for the doping control laboratory due to the many structurally similar steroids present in urinary matrices. This study aims to establish an innovative online two-dimensional high performance liquid chromatography (2D-HPLC) purification method for measuring the carbon isotope ratios (CIRs) and achieving the identification of the synthetic forms of these two endogenous anabolic androgenic steroids (EAASs). Initially, the one-dimensional chromatographic column was used to separate and purify endogenous reference compounds (ERCs), and the co-elution fluids containing PS and PSL were switched to a two-dimensional chromatographic column for further purification through an online transfer system. Then the purified compounds were analyzed using GC/C/IRMS after sample pretreatments. The results showed that the minimum detection concentration of PS and PSL reached 30 ng mL-1, and no isotope fractionation occurred during the entire collection and preparation process. This method has been validated with the WADA technical document and showed good sensitivity and selectivity, demonstrating its practical applicability for urine samples in doping control laboratories.

Concise synthesis of deuterium-labelled proanthocyanidin metabolite 5-(3',4'-dihydroxyphenyl)-γ-valerolactone as an analytical tool.

Deuterated proanthocyanidin metabolite 5-(3',4'-dihydroxyphenyl)-γ-valerolactone has been successfully produced. This metabolite is responsible for several proanthocyanidin protective effects in the field of cancer chemoprevention, skin wrinkle-prevention, and antimicrobials. The synthetic approach applied employs a short reaction sequence and allows the incorporation of four deuterium atoms on non-exchangeable sites, making it an attractive strategy to produce a stable isotopically labeled internal standard for quantitative mass spectrometry isotope dilution-based methods, as demonstrated by developing an LC-MS/MS method to quantify DHPV in urine samples. Overall, this efficient synthesis provides a valuable analytical tool for the study of the metabolic conversion of proanthocyanidins thus helping to investigate the biological effect and establishing the active dose of the key catabolite 5-(3',4'-dihydroxyphenyl)-γ-valerolactone.

In situ simultaneous measuring method for the determination of key processes of soil organic carbon cycling: Soil microbial respiration using laser spectrometry.

Soil microbial heterotrophic C-CO2 respiration is important for C cycling. Soil CO2 differentiation and quantification are vital for understanding soil C cycling and CO2 emission mitigation. Presently, soil microbial respiration (SR) quantification models are based on native soil organic matter (SOM) and require consistent monitoring of δ13C and CO2. We present a new apparatus for achieving in situ soil static chamber incubation and simultaneous CO2 and δ13C monitoring by cavity ring-down spectroscopy (CRDS) coupled with a soil culture and gas introduction module (SCGIM) with multi-channel. After a meticulous five-point inter-calibration, the repeatability of CO2 and δ13C values by using CRDS-SCGIM were determined, and compared with those obtained using gas chromatography (GC) and isotope ratio mass spectrometry (IRMS), respectively. We examined the method regarding quantifying SR with various concentrations and enrichment of glucose and then applied it to investigate the responses of SR to the addition of different exogenous organic materials (glucose and rice residues) into paddy soils during a 21-day incubation. The CRDS-SCGIM CO2 and δ13C measurements were conducted with high precision (<1.0µmol/mol and 1‰, respectively). The optimal sampling interval and the amount added were not exceeded 4 h and 200mg C/100g dry soil in a 1 L incubation bottle, respectively; the 13C-enrichment of 3%-7% was appropriate. The total SR rates observed were 0.6-4.2µL/h/g and the exogenous organic materials induced -49%-28% of priming effects in native SOM mineralisation. Our results show that CRDS-SCGIM is a method suitable for the quantification of soil microbial CO2 respiration, requiring less extensive lab resources than GC/IRMS.

Isotope effects accompanying δ2 H, δ18 O and δ17 O analyses of aqueous saline solutions using cavity ring-down laser spectroscopy.

The presence of substantial amounts of dissolved salts creates serious difficulties in isotope analyses of water samples using conventional isotope ratio mass spectrometry. Although nowadays laser-based instruments are increasingly used for this purpose, a comprehensive assessment of isotope effects associated with direct analyses of aqueous saline solutions using this technology is lacking. Here we report the results of laboratory experiments aimed at quantifying isotope effects associated with direct, δ2 H, δ18 O and δ17 O analyses of single-salt solutions and double-salt mixtures prepared with a water of known isotopic composition. Three single-salt solutions (NaCl, CaCl2 and MgSO4 ) and two double-salt mixtures (NaCl + CaCl2 and NaCl + MgSO4 ) were prepared and investigated for a wide range of molalities. The triple-isotope composition of the prepared solutions was analysed with the aid of a Picarro L2140-i Cavity Ring-Down Spectroscopy analyser. The NaCl and CaCl2 solutions revealed small negative salt effects, independent of molality and comparable with measurement uncertainty. The MgCl2 solution showed the highest salt effects, reaching saturated solution ca. +2.7‰ (2 H), -3.5‰ (18 O) and -1.7‰ (17 O). Salt effects for the double-salt mixtures generally mirrored the effects observed for the single-salt solutions. The observed salt effects are discussed in the context of processes occurring during the injection of the salt solutions into the vaporizer unit of the CRDS analyser. The presented study has demonstrated feasibility of direct, triple-isotope analyses of aqueous salt solutions using a Picarro L2140-i CRDS analyser for a broad range of salinities up to saturated conditions. Large uncertainties of 17 O-excess determinations for solutions forming hydrated salts preclude the use of this parameter for interpretation purposes.

Application of gas chromatography-stable isotope mass spectrometry to determine the oxygen isotope ratio of water in concentrated fruit juice

In this study, we developed a method to determine the authenticity of concentrated fruit juice using the δ18O ratio of water. We utilized the gas chromatography-isotope ratio mass spectrometry (GC-IRMS) technique along with the offline isotope exchange equilibrium method to pretreat and measure the oxygen isotopes in water in concentrated apple juice. This innovative approach uses anhydrous ethanol as a solvent to dissolve concentrated juices, thereby facilitating a more comprehensive reaction between the water in the sample and the CO2 standard gas. Additionally, the reaction vessel exhibits excellent air tightness, ensuring rapid equilibrium time and enabling accurate determination of δ18O in the water content of concentrated juice. The reaction vessel had excellent air tightness and rapid equilibration time, enabling accurate measurement of δ18O of water in concentrated fruit juice. The reaction vessel used had excellent air tightness and rapid equilibration time, ensuring accurate measurement of the δ18O of water in concentrated fruit juice. We analyzed 40 samples of concentrated apple juice and found that the range of δ18O values in the water of concentrated apple juice was between 1.29 ‰ and 7.59 ‰. Additionally, we used prepared sucrose syrup as a simulated sample and found that the δ18O distribution range was between −10.43 ‰ and −10.53 ‰. Furthermore, we conducted adulteration simulation experiments on randomly selected real samples and discovered a linear negative correlation between the adulteration ratio and the δ18O value of water in the samples.