New insights on the hydrochemistry, geothermometry, and isotopic characteristics of the hydrothermal groundwater of the SASS basin: case study of the Jérid geothermal field, Southern Tunisia.

Chemical and isotopic indicators were used to recognize the origin of hydrothermal groundwater, to assess the mineralization processes and groundwater quality, to identify the source of solutes and the likely mixing with cold, and elucidate the fluid geothermometry in the Jérid field of Southern Tunisia. The results show that the geothermal groundwater is neutral to slightly alkaline. They are characterized by SO4-Cl-Na-Ca water type. The dissolution of evaporates and pyrite-bearing rocks is the dominant mineralization process. The groundwater quality index indicates that the majority of samples are very hard and belong to poor to unsuitable for drinking classes. Applications and calculations of hydrogeochemical parameters, including SAR, %Na, PI, Kr, and MAR, showed that the majority of samples are unsuitable for agricultural practices. The human health risk was assessed based on hazard quotient and total hazard index through ingestion and dermal contact with iron-rich groundwater. The consumption of CI groundwaters does not present non-carcinogenic risk to adults and children. The δ18 O and δ2H signatures indicate that the geothermal groundwater was recharged by ocean precipitation during cold and wet paleoclimatic periods. The slight enrichment of oxygen-18 and deuterium contents suggests a limited mixing effect between geothermal water and cold groundwater within the same aquifer. This mixing effect is confirmed by the Na-K-Mg and the chloride-enthalpy diagrams. The K-Mg and SiO2 geothermometers provided fairly reliable reservoir temperature values, ranging between 69.6 and 99 °C. Calculated geothermal potential values, varying between 469 and 16987 kWth, which allow several applications such as domestic and agricultural heating.

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.

Tracking isotopically labeled oxidants using boronate-based redox probes.

Reactive oxygen and nitrogen species have been implicated in many biological processes and diseases, including immune responses, cardiovascular dysfunction, neurodegeneration, and cancer. These chemical species are short-lived in biological settings, and detecting them in these conditions and diseases requires the use of molecular probes that form stable, easily detectable, products. The chemical mechanisms and limitations of many of the currently used probes are not well-understood, hampering their effective applications. Boronates have emerged as a class of probes for the detection of nucleophilic two-electron oxidants. Here, we report the results of an oxygen-18-labeling MS study to identify the origin of oxygen atoms in the oxidation products of phenylboronate targeted to mitochondria. We demonstrate that boronate oxidation by hydrogen peroxide, peroxymonocarbonate, hypochlorite, or peroxynitrite involves the incorporation of oxygen atoms from these oxidants. We therefore conclude that boronates can be used as probes to track isotopically labeled oxidants. This suggests that the detection of specific products formed from these redox probes could enable precise identification of oxidants formed in biological systems. We discuss the implications of these results for understanding the mechanism of conversion of the boronate-based redox probes to oxidant-specific products.

GC-MS approach revealing hidden nitrite reactions in hemolysate using 18O-labelled nitrite and cysteine-based thiols.

In erythrocytes, nitric oxide (NO) and its autoxidation product nitrite undergo multiple reactions with hemoglobin species to form nitrate, Fe-nitrosyl hemoglobin (HbFe(II)NO), S-nitrosohemoglobin (HbCysß93SNO), and other intermediates including nitrito-methemoblobin HbFe(III)ONO, nitro-methemoblobin HbFe(III)NO2 and nitrous anhydride (N2O3). Here, we report a stable-isotope dilution GC-MS assay that allows studying reactions of nitrite in hemolysate. The method is based on the use of 18O-labelled nitrite in combination with l-cysteine or N-acetyl-l-cysteine ethyl ester and GC-MS measurement of unlabelled and labelled nitrite and nitrate species. This approach reveals reactions that are hidden at physiological nitrite concentrations.

Industrial separation of oxygen isotopes by oxygen distillation.

18 O-labeled water (Water-18 O) is a widely used starting material of 18 F-labeled diagnostic agents in positron emission tomography (PET). Conventionally, Water-18 O has been separated from other stable oxygen isotope species (16 O, 17 O) by water distillation or nitric oxide distillation. However, conventional methods are costly and may have safety issues. In 2004, we developed the first unit of our novel oxygen isotope separation process by cryogenic oxygen distillation to overcome these issues. To meet the needs of the market, we built a second unit in 2013 and a third in 2016. We are now operating three commercially viable separation units with a total capacity of 600 kg of Water-18 O per year.

An oxygen isotope chronometer for cellulose deposition: the successive leaves formed by tillers of a C4 perennial grass.

Multiannual time series of (palaeo)hydrological information can be reconstructed from the oxygen isotope composition of cellulose (δ18 OCel ) in biological archives, for example, tree rings, but our ability to temporally resolve information at subannual scale is limited. We capitalized on the short and predictable leaf appearance interval (2.4 d) of a perennial C4 grass (Cleistogenes squarrosa), to assess its potential for providing highly time-resolved δ18 OCel records of vapour pressure deficit (VPD). Plants grown at low (0.63 kPa) or high (1.58 kPa) VPD were swapped between VPD environments and exposed to the new environment for 7 d with simultaneous 13 CO2 labelling. Then, leaves were sampled by age/position along individual tillers. Five leaves at different developmental stages were growing simultaneously. The period of most-active leaf elongation, from 10 to 90% of final length, lasted 6.6 d, and ~80% of all carbon and oxygen incorporation in whole-leaf cellulose occurred within 7 d. Cellulose deposition stopped at (or shortly after) full leaf expansion. The direction of change, low-to-high or high-to-low VPD, had no differential effect on new oxygen and carbon incorporation in cellulose. Successive leaves produced by tillers of C. squarrosa provide a δ18 OCel record useful for reconstructions of short-term hydrological dynamics.

Seasonal changes in background levels of deuterium and oxygen-18 prove water drinking by harp seals, which affects the use of the doubly labelled water method.

The aim of this study was to monitor seasonal changes in stable isotopes of pool freshwater and harp seal (Phoca groenlandica) body water, and to study whether these potential seasonal changes might bias results obtained using the doubly labelled water (DLW) method when measuring energy expenditure in animals with access to freshwater. Seasonal changes in the background levels of deuterium and oxygen-18 in the body water of four captive harp seals and in the freshwater pool in which they were kept were measured over a time period of 1 year. The seals were offered daily amounts of capelin and kept under a seasonal photoperiod of 69°N. Large seasonal variations of deuterium and oxygen-18 in the pool water were measured, and the isotope abundance in the body water showed similar seasonal changes to the pool water. This shows that the seals were continuously equilibrating with the surrounding water as a result of significant daily water drinking. Variations in background levels of deuterium and oxygen-18 in freshwater sources may be due to seasonal changes in physical processes such as precipitation and evaporation that cause fractionation of isotopes. Rapid and abrupt changes in the background levels of deuterium and oxygen-18 may complicate calculation of energy expenditure by use of the DLW method. It is therefore strongly recommended that analysis of seasonal changes in background levels of isotopes is performed before the DLW method is applied on (free-ranging) animals, and to use a control group in order to correct for changes in background levels.

Evidence of the chemical reaction of (18)O-labelled nitrite with CO2 in aqueous buffer of neutral pH and the formation of (18)OCO by isotope ratio mass spectrometry.

Inorganic nitrite (NO2(-), ON-O(-) ←→ (-)O-NO) is the autoxidation product of nitric oxide (NO). Nitrite can also be formed from inorganic nitrate (ONO2(-)), the major oxidation product of NO in erythrocytes, by the catalytic action of bacterial nitrate reductase in gut and oral microflora. Nitrite can be reduced to NO by certain cellular proteins and enzymes, as well as in the gastric juice under acidic conditions. Hemoglobin, xanthine oxidoreductase and carbonic anhydrase (CA) have been reported to convert nitrite to NO. Renal CA isoforms are involved in the reabsorption of nitrite and may, therefore, play an important role in NO homeostasis. Yet, the mechanisms underlying the action of CA on nitrite are incompletely understood. The nitrate/nitrite system is regarded as a reservoir of NO. We have recently shown that nitrite reacts chemically with carbon dioxide (CO2), the regular substrate of CA. The present communication reports a stable isotope ratio mass spectrometry (IRMS) study on the reaction of NO2(-) and CO2 performed in 50 mM HEPES buffer of pH 7.4 at 37 °C. By using (18)O-labelled nitrite ((18)ON-O(-)/(-18)O-NO) and CO2 we observed formation of (18)O-labelled CO2. This finding is an unequivocal evidence of the chemical reaction of (18)ON-O(-)/(-18)O-NO with CO2. The reaction is rapid and involves nucleophilic attack of the negatively charged nitrite via one of its oxygen atoms on the partially positively charged CO2 molecule to form the putative intermediate (18)ON-O-CO2(-)/(-)O2C-(18)O-NO. The by far largest fraction of this intermediate decomposes back to (18)ON-O(-)/(-18)O-NO and CO2. A very small fraction of the intermediate, however, rearranges and finally decomposes to form (18)OCO and nitrite. This reaction is slower in the presence of an isolated erythrocytic CA isoform II. In summary, NO2(-), CO2 and CA are ubiquitous. The chemical reaction of NO2(-) with CO2 and its modulation by CA isoforms may play important roles in the transport of nitrite in red blood cells, the kidney and other cells and organs.

Comparison of carbon, nitrogen, and oxygen stable isotope ratios and mercury concentrations in muscle tissues of five beaked whale species and sperm whales stranded in Hokkaido, Japan.

We studied δ13C, δ15N and δ18O values, and total mercury (THg) concentrations in muscle samples from deep-sea predators - five beaked whale species and sperm whales - stranded along the coast of Hokkaido, in the north of Japan in 2010 and 2019. The δ13C, δ15N and δ18O values, THg concentrations, and body length (BL) of Stejneger's beaked whales were similar to those of Hubbs' beaked whales, which belong to the same genus. In contrast, δ13C values, THg concentrations, and BL of Sato's beaked whales were markedly different from those of Baird's beaked whales, which belong to the same genus. Stejneger's and Hubbs' beaked whales living around Hokkaido may compete in their ecological niches, whereas Sato's and Baird's beaked whales may segregate their ecological niches. Although Cuvier's beaked whales and sperm whales belong to different genera and their BLs were significantly different, their δ13C and δ15N values were similar, probably because they can dive and stay in deeper waters than other beaked whale species. The δ13C values in combined samples from all whales increased with increasing BL, probably owing to the larger whale species' dietary preference for squid. The δ13C values in combined samples from all whales were positively correlated with THg concentrations, whereas the δ15N values in the combined samples were negatively correlated. The δ18O values in combined samples from most whales tended to be positively correlated with THg concentrations. These correlations may be explained by a higher THg load from deep-sea feeding than from pelagic feeding and by a feeding shift towards lower trophic levels.

Isotope hydrology of the intermontane Elk Valley, British Columbia: an assessment of water resources around coal mining operations.

This study aimed to synthesise and interpret stable isotopic data (δ2H and δ18O) from various sources to understand the isotope hydrology around coal mine operations in Elk Valley, B.C., Canada. The data, including precipitation, groundwaters, seeps, and mine rock drains, were used to construct a local meteoric water line (LMWL) for the Elk Valley, evaluate the spatiotemporal isotopic composition of its groundwater, and assess mine seepage and mine rock drain discharge. The study revealed a robust LMWL relation (δ2H = 7.4 ± 0.2 · δ18O - 4.3 ± 4.1). The groundwater and seep data indicated a winter season bias and a north-south latitudinal gradient, suggesting rapid near-surface groundwater flow without significant post-precipitation evaporation. Porewater isotope samples from unsaturated mine rock piles (MRPs) showed site-specific evaporation patterns, potentially due to convective air flows or exothermic sulphide oxidation. This research revealed the influence of groundwater and meltwater on rock drain discharge. Based on evaporative mass balance calculations, MRPs seasonally contributed ca. 5 %(December base flow) and 22 % (snowmelt) to drain discharge. The findings underscore the value of stable isotope data collections in the Elk Valley to help better define and quantify the hydrology-hydrogeology, including a better understanding of evaporative conditions in MRPs.

The exploratory dataset of isotopic composition of different water sources across Kazakhstan.

This work presents the dataset of stable water isotopes of oxygen and hydrogen measured in water samples from different sources (precipitation, surface water, groundwater, tap water) across Kazakhstan from 2017 to 2018 and from 2020 to 2023. The dataset includes results on isotopic composition of 399 water samples, namely precipitation: event-based (n = 108), cumulative monthly (n = 22); surface water: lakes, reservoirs, brooks, rivers, channels (n = 175), groundwater: shallow and artesian groundwater, spring (n = 85), tapwater (n = 9). For each sample name of the source, location, latitude, longitude and date of sampling, measurement uncertainty (one standard deviation) are available. The samples were assessed by plotting the data in dual δ18O vs. δ2H isotope space with reference to values found in the published literature and fitting a linear regression equation for Astana (event) precipitation. Overall, this is the first dataset covering wide range of sources across Kazakhstan, which could be used by global and regional water resource assessments and studies such as tracing water sources, hydrograph separation and end-member analyses, isotope mass balance, evapotranspiration partitioning, residence time analysis and groundwater recharge.

A protocol for distilling animal body water from biological samples and measuring oxygen and hydrogen stable isotopes via cavity ring-down spectroscopy.

The application of stable isotope analysis (SIA) to the fields of ecology and animal biology has rapidly expanded over the past three decades, particularly with regards to water analysis. SIA now provides the opportunity to monitor migration patterns, examine food webs, and assess habitat changes in current and past study systems. While carbon and nitrogen SIA of biological samples have become common, analyses of oxygen or hydrogen are used more sparingly despite their promising utility for tracing water sources and animal metabolism. Common ecological applications of oxygen or hydrogen SIA require injecting enriched isotope tracers. As such, methods for processing and analyzing biological samples are tailored for enriched tracer techniques, which require lower precision than other techniques given the large signal-to-noise ratio of the data. However, instrumentation advancements are creating new opportunities to expand the applications of high-throughput oxygen and hydrogen SIA. To support these applications, we update methods to distill and measure water derived from biological samples with consistent precision equal to, or better than, ± 0.1 ‰ for δ17O, ± 0.3 ‰ for δ18O, ± 1 ‰ for δ2H, ± 2 ‰ for d-excess, and ± 15 per meg for Δ17O.

Topographic influence on ecohydrology in volcanic watersheds of the western Pacific monsoon area: evidence from water stable isotope composition of meteoric water, thermal water, and plants.

In Taiwanese volcanic watersheds, we investigated stable water isotopes in meteoric water, plants, and thermal water. Meteoric water exhibited a seasonal cycle, with heavier isotopes in winter and lighter ones in summer, especially in the southern region. The northern monsoon signal lagged the south by two weeks. In the Tatun mountains, young water fractions indicated prevalent old water sources. In the northern watershed, streamwater mainly came from the winter monsoon, while the southern one was influenced by alternating monsoons. Both indices indicated that winter plants depended on summer rainfall. Streamwater and plants had distinct sources in winter, supporting ecohydrological separation. Thermal spring water's d-excess helped identify water-rock interactions, with low d value signaling such interactions. The topographic wetness index showed a higher summer monsoon contribution to southern streamwater but a lower one to plants. The mean linear channel direction significantly affected the monsoon contribution fraction, with northeast-oriented channels vulnerable to northeastward winter monsoons. Finally, we developed a model illustrating hydrological processes on short and long timescales. Our findings enhance our understanding of hydrological disturbances' impact on water resources and ecosystems.

Origin and stability of pit lake water in Baiyinhua, Inner Mongolia, based on hydrochemistry and stable isotopes.

Isotope technology is widely used in geochemical mechanisms analysis; however, studies on the origin of pit lake water by isotopes in coal concentration areas in grassland are rare. In this study, 20 groups of water samples were collected, which were subjected to chemical analysis to determine the hydrogeochemical characteristics of pit lake water. The mechanisms of pit lake water formation and recharge-evaporation were ascertained through principal component analysis and the Rayleigh fractionation model. The results indicate that the phreatic water is least affected by evaporation, followed by confined water, surface water and pit lake water. The ionic composition of surface water, phreatic water and most of the confined water is mainly affected by leaching, some confined water can be recharged by surface or phreatic water; while the ionic composition of pit lake water is dominantly affected by evaporation (69.4 %) and is less affected by groundwater recharge (17.1 %) and human activities (11.5 %). The pit lake water is recharged by precipitation, phreatic water and the lateral runoff of confined water; however, the proportion of phreatic and confined water recharge is small. The evaporative loss of the pit lake water is 40-61 % of the initial water body.

The stable isotope hydrology of Sable Island, Nova Scotia, Canada with implications for evaluating the water budget of wild horses.

We investigated the stable isotope hydrology of Sable Island, Nova Scotia, Canada over a five year period from September, 2017 to August, 2022. The δ2H and δ18O values of integrated monthly precipitation were weakly seasonal and ranged from -66 to -15 ‰ and from -9.7 to -1.9 ‰, respectively. Fitting these monthly precipitation data resulted in a local meteoric water line (LMWL) defined by: δ2H = 7.22 ± 0.21 · δ18O + 7.50 ± 1.22 ‰. Amount-weighted annual precipitation had δ2H and δ18O values of -36 ± 11 ‰ and -6.1 ± 1.4 ‰, respectively. Deep groundwater had more negative δ2H and δ18O values than mean annual precipitation, suggesting recharge occurs mainly in the winter, while shallow groundwater had δ2H and δ18O values more consistent with mean annual precipitation or mixing of freshwater with local seawater. Surface waters had more positive values and showed evidence of isolation from the groundwater system. The stable isotopic compositions of plant (leaf) water, on the other hand, indicate plants use groundwater as their source. Fog had δ2H and δ18O values that were significantly more positive than those of local precipitation, yet had similar 17O-excess values. δ2H values of horsehair from 4 individuals lacked seasonality, but had variations typical to those of precipitation on the island. Differences in mean δ2H values of horsehair were statistically significant and suggest variations in water use may exist between spatially disparate horse communities. Our results establish an important initial framework for ongoing isotope studies of feral horses and other wildlife on Sable Island.

Isotopically labeled ozone: A new approach to elucidate the formation of ozonation products.

As ozonation becomes a widespread treatment for removal of chemicals of emerging concern from wastewater treatment plant effluents, there are increasing concerns regarding the formation of ozonation products (OPs), and their possible impacts on the aquatic environment and eventually human health. In this study, a novel method was developed that utilizes heavy oxygen (18O2) for the production of heavy ozone ([18O1]O2, [18O2]O1, [18O3]) to actively label OPs from oxygen transfer reactions. To establish and validate this new approach, venlafaxine with a well-described oxygen transfer reaction (tertiary amine -> N-oxide) was chosen as a model compound. Observed 18O/16O ratios in the major OP venlafaxine N-oxide (NOV) correlated with expected 18O purities based on tracer experiments. These results confirmed the successful labeling with heavy oxygen and furthermore demonstrate the potential to monitor NOV as an indicator of 18O/16O ratios during ozonation. As a next step, 18O/16O ratios were used to elucidate the formation mechanism of previously described OPs from sulfamethoxazole (SMX). Seven OPs were detected including the frequently described nitro-SMX, which was formed with a maximum yield of 3.2% (of initial SMX). With the successful labeling of six of the seven OPs from sulfamethoxazole, it was possible to confirm their previously proposed formation pathways, and to distinguish oxygen transfer from electron transfer reactions. 18O/16O ratios in OPs indicate that hydroxylation of the aromatic ring and formation of nitro-groups mostly follows oxygen transfer reactions, while electron transfer reactions initiate the formation of hydroxylamine and the abstraction of NH2 leading to catechol.

Spatial and meteorological controls of stable water isotope dynamics of precipitation in Kashmir Valley, Western Himalaya, India.

In the Himalayas, the lives and livelihoods of millions of people are sustained by water resources primarily depending on the moisture brought by Western Disturbances and Indian Summer Monsoon. In the present study, a network of 12 precipitation stations was established across the Kashmir Valley to understand the spatial and meteorological factors controlling precipitation isotopes. Temperature and relative humidity are dominant meteorological factors, whereas altitude, proximity to forest canopy, land use/land cover, windward and leeward sides of the mountains are the main physical factors influencing precipitation isotopes. The study suggests that the Mediterranean Sea and nearby water bodies along with continental recycling are the dominant sources of moisture from October to May, while the Arabian Sea, Bay of Bengal and continental recycling are the main sources of moisture from June to September. However, some precipitation events from October to May collect moisture from the Arabian Sea and some precipitation events from June to September collect moisture from the Mediterranean Sea. The occasional passage of Western Disturbances in summer merging with the Indian Summer Monsoon yields heavy to very heavy precipitation. The study provides a better understanding of complex spatial and meteorological phenomena controlling precipitation isotopes across the Western Himalayas.

Nitrate isotopes reveal N-cycled waters in a spring-fed agricultural catchment.

Nitrate stable isotopes provide information about nitrate contamination and cycling by microbial processes. The Fischa-Dagnitz (Austria) spring and river system in the agricultural catchment of the Vienna basin shows minor annual variance in nitrate concentrations. We measured nitrate isotopes (δ15N, δ18O) in the source spring and river up to the confluence with the Danube River (2019-2020) with chemical and water isotopes to assess mixing and nitrate transformation processes. The Fischa-Dagnitz spring showed almost stable nitrate concentration (3.3 ± 1.0 mg/l as NO3--N) year-round but surprisingly variable δ15N, δ18O-NO3- values ranging from +5.5 to +11.1‰ and from +0.5 to +8.1‰, respectively. The higher nitrate isotope values in summer were attributed to release of older denitrified water from the spring whose isotope signal was dampened downstream by mixing. A mixing model suggested denitrified groundwater contributed > 50 % of spring discharge at baseflow conditions. The isotopic composition of NO3- in the gaining streams was partly controlled by nitrification during autumn and winter months and assimilation during the growing season resulting in low and high δ15N-NO3- values, respectively. NO3- isotope variation helped disentangle denitrified groundwater inputs and biochemical cycling processes despite minor variation of NO3- concentration.

New insights into diffusive kinetic fractionation during liquid condensation under supersaturated environment: an alternative approach for isotope tagging of ground-level water vapour.

Stable water isotopes in ground-level vapour are key to estimating water exchange between geospheres. Their sampling, however, is limited to laser-absorption spectrometers and satellite observations, having inherent shortcomings. This study investigates diffusive kinetic fractionation during liquid condensation under supersaturated environment, providing a cost-effective, reliable way of sampling ground-level vapour isotopes (18O, 2H). Experiments were undertaken at three locations in India with 'liquid' samples collected from condensation of ambient air at 0°C. Simultaneously, pristine 'vapour' was sampled via cryogenic-trapping using liquid nitrogen-alcohol slush at -78°C. The 'liquid' condensed under supersaturation was progressively more depleted in 18O, and less enriched in 2H than expected under equilibrium fractionation, with an increasing degree of supersaturation expressed as saturation index (Si). This study revealed: (1) Si, molecular density, Rh, T together control the extent of isotopic kinetic fractionation. (2) The presence of diffusive concentration gradient inhibits the flow of heavier isotopes during liquid condensation. (3) The stochastic nature of the process cannot be explained using a physics-based model alone. The artificial neural network model is hence deployed to sample δ18O (δ 2H) within -0.24 ± 1.79‰ (0.53 ± 11.23 ‰) of true value. (4) The approach can be extended to ground-validate isotope-enabled general circulation models and satellite observations.

Nitrate isotopes (δ15N, δ18O) in precipitation: best practices from an international coordinated research project.

Stable isotope ratios of nitrogen and oxygen (15N/14N and 18O/16O) of nitrate (NO3-) are excellent tracers for developing systematic understanding of sources, conversions, and deposition of reactive atmospheric nitrogen (Nr) in the environment. Despite recent analytical advances, standardized sampling of NO3-) isotopes in precipitation is still lacking. To advance atmospheric studies on Nr species, we propose best-practice guidelines for accurate and precise sampling and analysis of NO3- isotopes in precipitation based on the experience obtained from an international research project coordinated by the International Atomic Energy Agency (IAEA). The precipitation sampling and preservation strategies yielded a good agreement between the NO3- concentrations measured at the laboratories of 16 countries and at the IAEA. Compared to conventional methods (e.g., bacterial denitrification), we confirmed the accurate performance of the lower cost Ti(III) reduction method for isotope analyses (15N and 18O) of NO3- in precipitation samples. These isotopic data depict different origins and oxidation pathways of inorganic nitrogen. This work emphasized the capability of NO3- isotopes to assess the origin and atmospheric oxidation of Nr and outlined a pathway to improve laboratory capability and expertise at a global scale. The incorporation of other isotopes like 17O in Nr is recommended in future studies.