Evaluation of lipid extraction methods for fatty acid quantification and compound-specific δ13C and δ2Hn analyses.

Lipids, with fatty acids (FA) as a crucial subset, have become a focal point for diverse medical, physiological, and ecological studies. However, a comprehensive assessment of the various pre-analytical FA extraction methods published in the scientific literature remains lacking. In this study, we examined the efficacy of seven well-established sample preparation methods, specifically focusing on their effectiveness in total lipid and fatty acid extraction and their impact on compound-specific stable hydrogen (δ2H) and carbon (δ13C) isotope values. We also considered the repercussions of FA removal efficacy on residual bulk tissue δ2Hn analysis, because lipids typically have low δ2H values. Our findings showed that in most cases chloroform-based extraction methods outperformed those without chloroform. While discrepancies were not as evident for smaller organisms, such as plankton, marked variations were discernible in the extraction efficiencies for muscle and liver samples, which was also manifested in the residual bulk tissue δ2Hn results. Notably, most extraction methods had little effect on specific δ13C or δ2H isotope values of FA; instead, an emphasis should be on using an extraction method that achieves optimal baseline peak separation of the chromatograms for C and H isotope measurements.

Radiocarbon integrity of dissolved inorganic carbon (DIC) samples stored in plastic and glass bottles: implications for reliable groundwater age dating.

Various approaches based on the natural variations of carbon isotopes (14C and 13C) in dissolved inorganic carbon (DIC) are routinely used to study groundwater dynamics and to estimate recharge rates by deriving groundwater ages. However, differences in 14C activities in groundwater samples collected repeatedly from the same wells and discordantly young 14C groundwater ages compared to noble gases led some authors to question the validity of radiocarbon dating. Poor sampling protocols and storage effects (14C contamination) for radiocarbon analysis are a critical factor in explaining age determination discrepancies. We evaluated the impact of storage protocols on carbon isotope exchange with atmospheric carbon dioxide by comparing glass versus standard plastic field sampling bottles for various storage times before radiocarbon and 13C analyses. The 14C bias after 12 months in pre-evacuated glass vials was minimal and within analytical precision. However, storage of DIC samples in plastic sampling bottles led to marked changes in 14C and 13C contents (up to ∼15 pmC and ∼ 5 ‰, respectively, after 12 months), meaning contamination led to younger groundwater age estimations than it should have been. Protocols for sampling and storing DIC samples for radiocarbon using pre-evacuated glass bottles help avoid atmospheric 14CO2 contamination and microbial activity.

Protocols for sample preparation and compound-specific stable-isotope analyses (δ2H, δ13C) of fatty acids in biological and environmental samples.

Compound-specific stable-isotope analysis (CSIA) of fatty acids is a powerful tool to better understand the trophic transfer of fatty acids and their biochemical fate in and across ecosystems, including tracing animal migration and understanding physiological processes. The non-exchangeable nature of C-H bonds in acyl chains, hydrogen (δ2H) and carbon (δ13C) stable-isotope values of fatty acids (FA) provide independent information about the origins of fatty acids. Several technical obstacles must be overcome to ensure accurate and reproducible measurements of FA-CSIA can be made. This protocol describes the sample preparation process for successful stable-isotope analyses of fatty acids obtained from environmental and biological samples. Numerous techniques for the preanalytical processing of fatty acid samples are available, and these often have minimal impact on δ values. Here, we provide an in-depth guide detailing our well-established laboratory protocols, ranging from the initial sample preparation, lipid extraction, and transmethylation to the instrumental arrangement, data collection, and analysis.•Protocol from obtaining a sample to standardized fatty acid specific δ2H and δ13C values.•Separate GC analysis procedures for C and H are recommended for optimal performance.

Global and local meteoric water lines for δ17O/δ18O and the spatiotemporal distribution of Δ'17O in Earth's precipitation.

Recently, δ17O and its excess (Δ'17O) have become increasingly significant "triple-oxygen-isotope" indicators of distinctive hydrological processes in hydrology and climatology. This situation mirrors the research regarding δ18O and δ2H in the 1960s towards a solid theoretical base and a surge in application examples and field studies worldwide. Currently, systematic global measurements for δ17O in precipitation are still lacking. As a result, attempts have been made to define a Global δ17O/δ18O Meteoric Water Line (GMWL), often by using regional or local datasets of varying systematicity. Different definitions of the global reference slope (λref) for determining Δ'17O values have been proposed, by ongoing debate around a proposed consensus value of 0.528. This study used worldwide samples archived in the IAEA Global Network of Isotopes in Precipitation (GNIP) to (a) derive a δ17O/δ18O GMWL based on four-year monthly records from 66 GNIP stations, (b) formulate local δ17O/δ18O meteoric water lines (LMWL) for these stations' areas, and (c) evaluate regional and seasonal variations of Δ'17O in precipitation. The GMWL for δ17O/δ18O was determined to be δ'17O = 0.5280 ± 0.0002 δ'18O + 0.0153 ± 0.0013, in keeping with the consensus value. Furthermore, our results suggested that using a line-conditioned 17O-excess is a viable alternative over the global λref in the context of regional hydrology and paleoclimatology interpretations; however, without challenging the global λref as such.

Assessing the performance of international laboratories analysing the stable isotope composition (δ15 N, δ18 O, δ17 O) of nitrate in environmental waters.

RATIONALE: Stable-isotope analyses of nitrate (NO3 - ) in various water sources are crucial for understanding nitrogen pollution and its impact on aquatic ecosystems. We evaluated the accuracy and precision of stable-isotope analyses of nitrate conducted by international laboratories. METHODS: Six samples with nitrate (2 mg L-1 NO3 - -N) were sent to 47 laboratories. The NO3 - had a 30-50 ‰ range of δ values for δ15 N, δ18 O and δ17 O. One blind duplicate evaluated reproducibility and the effect of water δ18 O. Laboratories used diverse methods to convert nitrate to N2 O, N2 , CO or O2 for stable-isotopic measurements (microbial, cadmium, titanium and elemental analysis) and isotope-ratio mass spectrometry or laser-based technologies. RESULTS: Thirty-six international laboratories (83 %) reported results; however, 23 % did not analyze the test samples due to technical difficulties. Of the reporting laboratories, 79 % and 84 % produced accurate δ15 N and δ18 O results falling within ±0.8 ‰ and ±1.1 ‰ of the benchmark values, respectively. Three laboratories produced only outliers. The duplicate revealed most laboratories gave internally reproducible results at appropriate analytical precision. For δ17 O, six laboratories reported results, but 67 % could not reproduce results within their claimed analytical measurement precision. One complication is a lack of nitrate reference materials for δ17 O. CONCLUSIONS: Analyst experience contributed to better performance, and underperformance was from compromised standards or inappropriate δ range of working reference materials. The stable isotope community must develop new nitrate reference materials for δ15 N spanning -20 ‰ to +80 ‰ and new materials for δ17 O.

Response of stable isotopes (δ2H, δ13C, δ15N, δ18O) of lake water, dissolved organic matter, seston, and zooplankton to an extreme precipitation event.

Lake ecosystems process and cycle organic substrates, thus serving as important bioreactors in the global carbon cycle. Climate change is predicted to increase extreme weather and precipitation events that can flush nutrients and organic matter from soils to streams and lakes. Here we report changes in stable isotopes (δ2H, δ13C, δ15N, or δ18O) of water, dissolved organic matter (DOM), seston, and zooplankton in a subalpine lake at short time resolution following an extreme precipitation event between early July to mid-August 2021. Water from excess precipitation and runoff remained in the lake epilimnion and coincided with increasing δ13C values of seston (-30 ‰ to -20 ‰), due to the input of carbonates and terrestrial organic matter. Particles settled into deeper lake layers after two days and contributed to the uncoupling of C and N cycling as the lake responded to this extreme precipitation event. Following the event, there was an increase in bulk δ13C values of zooplankton (from -35 ‰ to -32 ‰). Throughout this study, δ13C values of DOM remained stable throughout the water column (-29 ‰ to -28 ‰), while large isotopic fluctuations in DOM δ2H (-140 ‰ to -115 ‰) and δ18O (+9 ‰ to +15 ‰) values suggested DOM relocation and turnover. Integrating isotope hydrology, ecosystem ecology, and organic geochemistry offers an element-specific, detailed approach to investigating the impact of extreme precipitation events on freshwater ecosystems and particularly aquatic food webs.

Automated rapid triple-isotope (δ15N, δ18O, δ17O) analyses of nitrate by Ti(III) reduction and N2O laser spectrometry.

The nitrogen and oxygen (δ15N, δ18O, δ17O) stable isotopic compositions of nitrate (NO3-) are crucial tracers of nutrient N sources and dynamics in aquatic and atmospheric systems. Methods to reduce aqueous NO3- to N2O gas (microbial or Cd method) before 15N and 18O isotope analyses require multi-step conversion or toxic chemicals, and 17O in N2O cannot be disentangled by IRMS due to isobaric interferences. This technical note describes the automation of the stable-isotope analyses of nitrate by coupling the new Ti method with a headspace autosampler and an N2O triple-isotope laser analyzer based on off-axis integrated cavity output spectroscopy. The automation yielded accurate and precise results for routine determinations of δ15N, δ18O, and δ17O values for aqueous nitrate in environmental waters. Systematic corrections were required for cavity pressure, N2O concentration and water vapour content to obtain the highest precision for all three isotopic ratios. For the first time, an automated laser-based system facilitates routine low-cost triple isotope analyses in studies where high-temporal resolution isotope analyses of NO3- are required but have been, until now, cost-prohibitive and time-consuming (e.g. atmospheric N pollution).

Balancing precision and throughput of δ 17O and Δ'17O analysis of natural waters by Cavity Ringdown Spectroscopy.

δ 17O and Δ'17O are emerging tracers increasingly used in isotope hydrology, climatology, and biochemistry. Differentiating small relative abundance changes in the rare 17O isotope from the strong covariance with 18O imposes ultra-high precision requirements for this isotope analysis. Measurements of δ 17O by Cavity Ringdown Spectroscopy (CRDS) are attractive due to the ease of sample preparation, automated throughput, and avoidance of chemical conversions needed for isotope-ratio mass spectrometry. However, the CRDS approach requires trade-offs in measurement precision and uncertainty. In this protocol document, we present the following:•New analytical procedures and a software tool for conducting δ 17O and Δ'17O measurements by CRDS.•Outline a robust uncertainty framework for Δ'17O determinations.•Description of a CRDS performance framework for optimizing throughput, instrumental stability, and Δ'17O measurement precision and accuracy.

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.

Reproducible measurements of the δ2H composition of non-exchangeable hydrogen in complex organic materials using the UniPrep2 online static vapour equilibration and sample drying system.

Non-exchangeable hydrogen-isotope (δ 2Hn) measurements of complex organic samples are used in forensics to determine sample authenticity, traceability, and provenance. However, δ 2Hn assays of organics are usually complicated by uncontrolled "exchangeable hydrogen" and residual moisture contamination; hence, δ 2Hn assays are persistently incomparable amongst laboratories. We introduce a revised technical solution (UniPrep2) to control hydrogen-isotope exchange and for robust online sample drying and vapour equilibration. The UniPrep2 device is coupled to a high-temperature thermochemical elemental analyser and continuous-flow isotope-ratio mass spectrometer. This technical solution empowers isotope analysts to:•Conduct reproducible controlled vapour equilibrations of complex organic samples and standards to determine the δ 2Hn values by controlling hydrogen-isotope exchange.•Conduct online vacuum-oven evacuation with extensive helium drying without exposure to air to reabsorb or exchange hydrogen with ambient water vapour. The protocol describes the operation of the UniPrep2 device and the step-by-step procedures needed to obtain accurate and precise δ 2Hn values for a wide range of organic sample types. Two analytical approaches are described in detail; the Dual-Vapour Equilibration (DVE) approach, intended for determining δ 2Hn for a complex organic environmental sample where matrix equivalent H isotope reference materials are not available, and the Comparative Equilibration (CE) approach, which is intended for routine high-throughput analyses of complex organic samples where at least two matrix-equivalent organic isotope reference materials with consensus δ 2Hn values are being used. These standard operating procedures are envisioned to be a sound basis for advancing hydrogen-isotope analysis for different organic environmental matrices and studies.

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.

Quantifying denitrification in a field-scale bioremediation experiment.

Nitrate (NO3-) in mine waste rock derived from undetonated NH4NO3 can contaminate receiving waters. An in-situ bioremediation experiment was conducted at a coal mining operation in Elk Valley, British Columbia, Canada to remediate NO3- from large volumes of mine water. Over the test period (201 d), 5000 to 7500 m3 d-1 of NO3--rich (mean concentration 22 mg N L-1) mine water was injected into saturated waste rock along with methanol, nutrients, and a conservative tracer (Br-). Complete denitrification (<0.5 mg N L-1) was recorded in monitoring wells located 38 m from the injection wells after 114 to 141 d of operation. Plots of δ15N- and δ18O-NO3- versus NO3--N concentrations for monitoring wells yielded isotopic enrichment factors (ε) for δ15N- and δ18O-NO3- of -25.7 and -13.2 ‰ for high C/C0 NO3- concentrations (>10.5 mg N L-1) and -5.5 and -3.6 ‰ for lower C/C0 values. The fraction of NO3- denitrified (Dp) calculated using bi-linear ε values for δ15N- and δ18O reproduced the Dp determined independently using a conservative tracer indicating that stable isotope tracers of the NO3- reducing processes in bioremediation are invaluable to determine Dp. Based on the success of this ongoing bioremediation experiment, the technology is being applied at other sites.

Assessment of Compound-Specific Fatty Acid δ13C and δ2H Values to Track Fish Mobility in a Small Sub-alpine Catchment.

Methods for identifying origin, movement, and foraging areas of animals are essential for understanding ecosystem connectivity, nutrient flows, and other ecological processes. Telemetric methods can provide detailed spatial coverage but are limited to a minimum body size of specimen for tagging. In recent years, stable isotopes have been increasingly used to track animal migration by linking landscape isotope patterns into movement (isoscapes). However, compared to telemetric methods, the spatial resolution of bulk stable isotopes is low. Here, we examined a novel approach by evaluating the use of compound-specific hydrogen and carbon stable isotopes of fatty acids (δ2HFA and δ13CFA) from fish liver, muscle, brain, and eye tissues for identifying site specificity in a 254 km2 sub-alpine river catchment. We analyzed 208 fish (European bullhead, rainbow trout, and brown trout) collected in 2016 and 2018 at 15 different sites. δ13CFA values of these fish tissues correlated more among each other than those of δ2HFA values. Both δ2HFA and δ13CFA values showed tissue-dependent isotopic fractionation, while fish taxa had only small effects. The highest site specificity was for δ13CDHA values, while the δ2H isotopic difference between linoleic acid and alpha-linolenic acid resulted in the highest site specificity. Using linear discrimination analysis of FA isotope values, over 90% of fish could be assigned to their location of origin; however, the accuracy dropped to about 56% when isotope data from 2016 were used to predict the sites for samples collected in 2018, suggesting temporal shifts in site specificity of δ2HFA and δ13CFA. However, the predictive power of δ2HFA and δ13CFA over this time interval was still higher than site specificity of bulk tissue isotopes for a single time point. In summary, compound-specific isotope analysis of fatty acids may become a highly effective tool for assessing fine and large-scale movement and foraging areas of animals.

High spatial resolution prediction of tritium (3H) in contemporary global precipitation.

Tritium (3H) in Earth's precipitation is vigilantly monitored since historical nuclear bomb tests because of radiological protection considerations and its invaluable role as a tracer of the global water cycle in quantifying surface, groundwater, and oceanic fluxes. For hydrological applications, accurate knowledge of 3H in contemporary local precipitation is prerequisite for dating of critical zone water and calibrating hydrogeologic transport and groundwater protection models. However, local tritium input in precipitation is hard to constrain due to few 3H observation sites. We present new high-spatial resolution global prediction maps of multi-year mean 3H in contemporary "post-bomb" (2008-2018) precipitation by using a robust regression model based on environmental and geospatial covariates. The model accurately predicted the mean annual 3H in precipitation, which allowed us to produce global 3H input maps for applications in hydrological and climate modelling. The spatial patterns revealed natural 3H in contemporary precipitation sufficient for practical hydrological applications (1-25 TU) but variable across continental regions and higher latitudes due to cumulative influences of cyclical neutron fluxes, stratospheric inputs, and distance from tropospheric moisture sources. The new 3H maps provide a foundational resource for improved calibration of groundwater flow models and critical zone vulnerability assessment and provides an operational baseline for quantifying the potential impact of future anthropogenic nuclear activities and hydroclimatic changes.

Assessment of rapid low-cost isotope (δ15 N, δ18 O) analyses of nitrate in fruit extracts by Ti(III) reduction to differentiate organic from conventional production.

RATIONALE: The isotopic composition (δ15 N, δ18 O) of nitrate in fruits and vegetables differentiates organic from conventional food production practices. Organic systems do not use synthetic nitrate fertilizers high in 18 O and low in 15 N and thereby help reveal producers' fertilization claims. Isotope analyses of nitrate extracted from fruits and vegetables are done by bacterial reduction which is costly and by specialized laboratories. Rapid, low-cost methods are needed to promulgate nitrate isotope analyses of food products to support organic food product certification and to verify the authenticity of production claims. METHODS: Fresh strawberry samples were obtained from certified organic and conventional growers in Andalucía, Spain. We applied a new, rapid, one-step Ti(III) reduction method to convert the nitrate from strawberry extracts to N2 O gas for headspace isotope analyses using isotope-ratio mass spectrometry. Using the Ti(III) reduction method, 70 samples, controls and references were prepared and analyzed for NO3 - , δ15 N and δ18 O per 48 h. We also analyzed extracts and solids for anions and cations and for bulk δ15 N for multivariate chemometric evaluation. RESULTS: The Ti(III)-based isotope analyses of nitrate in strawberry extracts revealed clear differentiation between organic and conventional production with mean δ18 O and δ15 N values of +18.3 ± 1.2 ‰ and +17.6 ± 1.2 ‰ versus +28.2 ± 4.5 ‰ and +14.9 ± 3.0 ‰, respectively. The δ15 N of strawberry dry mass differed slightly (+3.0 ± 1.4 ‰ versus +4.0 ± 1.4 ‰) between organic and conventional samples, respectively. Chemometric analyses of nitrate isotopes and extract chemistry revealed that the δ18 O of nitrate along with δ15 N and Ca2+ fully differentiated organic from conventional strawberry production. CONCLUSIONS: Our results show the Ti(III) reduction method provides a new low-cost and rapid analytical method to facilitate compound-specific δ15 N and δ18 O isotope analyses of nitrate in selected fruit types, and likely other food products, for the purposes of assessing nitrate fertilization practices of organic versus conventional production claims and to support authenticity investigations.

Stable isotopes in global lakes integrate catchment and climatic controls on evaporation.

Global warming is considered a major threat to Earth's lakes water budgets and quality. However, flow regulation, over-exploitation, lack of hydrological data, and disparate evaluation methods hamper comparative global estimates of lake vulnerability to evaporation. We have analyzed the stable isotope composition of 1257 global lakes and we find that most lakes depend on precipitation and groundwater recharge subsequently altered by catchment and lake evaporation processes. Isotope mass-balance modeling shows that ca. 20% of water inflow in global lakes is lost through evaporation and ca. 10% of lakes in arid and temperate zones experience extreme evaporative losses >40 % of the total inflow. Precipitation amount, limnicity, wind speed, relative humidity, and solar radiation are predominant controls on lake isotope composition and evaporation, regardless of the climatic zone. The promotion of systematic global isotopic monitoring of Earth's lakes provides a direct and comparative approach to detect the impacts of climatic and catchment-scale changes on water-balance and evaporation trends.

Performance of low-cost stainless-steel beverage kegs for long-term storage integrity and easy dispensing of water isotope (δ18 O, δ2 H) reference materials.

RATIONALE: A widespread problem observed in global water isotope (δ18 O, δ2 H) proficiency tests is compromised working reference materials due to storage-dispensing evaporation effects. Proper storage requires no evaporation or leakage, which causes isotopic drift and bias. Surveys by the International Atomic Energy Agency (IAEA) show most isotope laboratories use glass or plastic bottles to store working reference materials, with frequent opening and closings that pose evaporation risks. Practical small (ca. 2-5 L) storage-dispensing solutions free of air exposure, evaporation, and leakage are needed. We also tested several smaller-scale bottles for day-to-day aliquots. METHODS: We tested low-cost, conveniently sized (4 L) adaptations of a common stainless-steel beverage keg with a liquid dispenser, with minor modifications to facilitate low-flow dispensing and pressurization (1-2 bar) with Ar or N2 . We tested three kegs (100%, 75%, 50% initial fills) for a 2-year period along with monthly dispensing to assess long-term storage viability for maintaining δ18 O and δ2 H integrity and dispensing, and day-to-day aliquot bottles for 6 months. RESULTS: Test results showed these small keg storage systems fully maintained the isotopic integrity of water over the 2-year testing period with no trend in the isotopic data that would reveal evaporative loss or leakage (e.g., pressure or mass loss) regardless of starting fill level. However, evaporated water in the outlet tube assembly must be eliminated by discarding 15-20 mL before dispensing into appropriate daily-use laboratory standard bottles (30-100 mL). Glass bottles for daily aliquots showed good integrity properties, but only if their fill level was >50%. CONCLUSIONS: The use of a low-cost pressurized metal beverage keg dispensing system provides a robust solution to enable laboratories to maintain the integrity of their water isotope working reference materials over several years.

Comparative evaluation of 2H- versus 3H-based enrichment factor determination on the uncertainty and accuracy of low-level tritium analyses of environmental waters.

Analysis of low-level tritium (3H) in environmental waters requires pre-concentration using electrolytic enrichment prior to decay counting. Accurate and precise electrolytic enrichment factors (EF) are required to determine the sample's environmental 3H concentration. Two methods are used to determine EFs: i) the Spike Proxy Method (SPM) and ii) the Deuterium Method (DM) with each having several modalities. We conducted a comparative assessment of four EF strategies using 250 mL and 500 mL electrolytic enrichment of three low-level 3H proficiency water standards (0.5-7 TU) to see which strategy gave the most accurate 3H results based on z- and Zeta-scores. Our comparative evaluation revealed the DM offers consistently superior 3H results, with more precise EF determinations compared to the three SPM strategies. The DM gave the best z-scores with an EF relative combined uncertainty of about 0.5‰ and a negligible contribution to the overall uncertainty budget due to the EF determination. Moreover, the DM can improve productivity by eliminating the spike and gravimetric procedures from routine analyses and can give rapid cell enrichment performance feedback prior to decay counting. We recommend low-level tritium laboratories consider adopting the DM into their 3H sample enrichment and analysis operations.

Compound-specific stable hydrogen isotope (δ2 H) analyses of fatty acids: A new method and perspectives for trophic and movement ecology.

RATIONALE: Compound-specific stable isotope analysis (CSIA) is a powerful tool for a better understanding of trophic transfer of dietary molecules in and across ecosystems. Hydrogen isotope values (δ2 H) in consumer tissues have potential to more clearly distinguish dietary sources than 13 C or 15 N values within and among habitats, but have not been used at the fatty acid level for ecological purposes. METHODS: Here we demonstrate a new online high-capacity gas chromatography-isotope ratio mass spectrometry technique (2 H-CSIA) that offers accurate and reproducible determination of δ2 H values for a range of fatty acids from organisms of aquatic food webs. RESULTS: We show that lipid extracts obtained from aquatic sources, such as biofilms, leaves, invertebrates, or fish muscle tissue, have distinctive δ2 H values that can be used to assess sources and trophic interactions, as well as dietary allocation and origin of fatty acids within consumer tissue. CONCLUSIONS: The new 2 H-CSIA method can be applied to evaluate sources and trophic dynamics of fatty acids in organisms ranging from food web ecology to migratory connectivity.

Principles and uncertainties of 14C age estimations for groundwater transport and resource evaluation.

Radiocarbon (14C) is useful for estimating groundwater ages for transport and water resource exploitation assessment. If the 14C content of dissolved inorganic carbon (14CDIC) is known, the age of groundwater can be estimated by applying a radiocarbon decay equation combined with an appropriate geochemical correction model. However, age determinations are subject to uncertainties caused by parameters which need to be estimated or assumed. Here, we discuss the principles of 14C-based groundwater age estimations and the corrections and errors that affect age determinations differently. Generally, the two factors that impact the results of 14C groundwater age are Type-1 and Type-2 errors. Type-1 errors are pulse-type changes on derived groundwater ages that are independent of the water age. Type-2 errors cause gradual changes on derived groundwater 14C ages that depend on the water age. The cumulative impact of these errors substantively reduces the accuracy and confidence of 14C age determinations and corrections. When using 14C for groundwater age, consideration of both error types along with the use of samples having a range of 14CDIC contents helps practitioners recognize and minimize 14C age uncertainty, especially for groundwater ages of <1000 and >30,000 years B.P.