A new approach to determining the content and 15N abundance of total dissolved nitrogen in aqueous samples: TOC analyser-QMS coupling

The standard method for determining the 15N abundance of total dissolved nitrogen (TDN) in aqueous samples (e.g., soil leachate, sewage, urine) is currently Kjeldahl digestion followed by steam distillation or diffusion to isolate the ammonium, and then 15N measurement using IRMS. However, this technique is both time-consuming and laborious. One way of overcoming these disadvantages could be to couple a TOC analyser to determine the TDN with a sufficient quadrupole MS to determine the 15N abundance. The high TOC analyser (Elementar Analysensysteme Hanau, Germany), which catalytically oxidises the sample's total nitrogen with a high, constant yield to nitrogen monoxide (NO), appeared particularly suitable. The quadrupole-MS ESD 100 (InProcess Instruments Bremen, Germany) proved to be a suitable mass spectrometer for the 15N determination of NO. This combination of instruments was found to provide a workable method in numerous measurements of standard and actual samples. The detection limit concerning the N amount required per analysis is 2 microg, corresponding to an N concentration of 0.7 mg/l in a maximum sample volume of 3ml. Depending on the N concentration, 15N abundances starting from 0.5 at.% can be measured with the required precision of better than 3% (simple standard deviation). For example, measuring the abundance of 0.5 at.% requires about 50 microg N, whereas for 1 at.% or more only about 5 microg N is needed per analysis.     

Determination of (15)N in (15)N-enriched nitrite and nitrate in aqueous samples by reaction continuous flow quadrupole mass spectrometry

The (15)N tracer method is the most suitable method for studying complex N transformation processes in microbiology and biochemistry. It entails the constant determination of the (15)N abundance of the inorganic nitrogen (N) compounds nitrite and nitrate. However, (15)N analytical methods are time-consuming, difficult to automate, and require at least 10 μg of N per determination. An additional obstacle in the case of nitrite is that it usually only occurs in very small amounts (ppb) dwarfed by much larger quantities of nitrate (ppm). More useful is an approach in which the N compound is selectively converted into a gaseous form suitable for direct measurement by mass spectrometry. By using this 'reaction continuous-flow mass spectrometry' (R/CFMS) we developed methods for the (15)N determination of nitrite and nitrate from tracer experiment samples, i.e. artificially enriched in (15)N. Because both methods are based on the same principle, one continuous flow setup connected directly to a quadrupole mass spectrometer for all determinations was used. Nitrite and nitrate are reduced to NO by iodide and titanium(III) chloride, respectively. The technique developed ensures a precision of relative standard deviation /=1 at.% are to be measured for nitrite and nitrate, respectively. Copyright 1999 John Wiley & Sons, Ltd.     

Determining the content and 13C abundance of total dissolved carbon in water samples by TOC analyser-mass spectrometer coupling

A combined system consisting of a TOC analyser connected to a quadrupole MS was recently described as a way of measuring the N content and the ¹⁵N abundance of total dissolved nitrogen in aqueous samples. This work examines whether this combination of instruments can also be used for the ¹³C determination of the total dissolved carbon in aqueous samples. A level of precision good for ¹³C-enriched samples was achieved with a relative standard deviation of <3%. By using an isotope ratio MS instead of the quadrupole MS employed here, TOC-MS coupling also ought to be suitable for determining natural ¹³C abundances.     

Methodische Untersuchungen zum 15N-ConFlo-IRMS-System

After installing a C-GC-IRMS-System (Elemental analyzer EA 1108 CHN + IRMS MAT 252) some problems arose with respect to the external precision of the δ¹⁵N values (standard deviation approx. 1‰ at < 50 μg N) after changeover from ¹³C to ¹⁵N measurements.

Attempts were made to determine the reasons for these effects and to eliminate them by changes of the Carlo Erba on-line combustion system (EA) and the split.

Non-quantifiable contributions, e.g., air nitrogen and/or N₂ in carrier gas helium were reduced by a blank correction.

Using Cyclohexanone-2, 4-dinitrophenylhydrazone it was shown that under the same conditions and after a common blank correction with small and large amounts of samples (60-650 μg, 20% N) an external precision of 0.2‰ (standard deviation, 10 cycles) can be reached.     

Using the natural 15N abundance to assess the main nitrogen inputs into the sand dune area of the North-Western Negev desert (ISRAEL)

The variation of the natural ¹⁵N abundance is often used to evaluate the origin of nitrogen or the pathways of N input into ecosystems. We tried to use this approach to assess the main input pathways of nitrogen into the sand dune area of the north-western Negev Desert (Israel). The following two pathways are the main sources for nitrogen input into the system:

1. Biological fixation of atmospheric nitrogen by cyanobacteria present in biological crusts and by N₂-fixing vascular plants (e.g. the shrub Retama raetam);

2. Atmospheric input of nitrogen by wet deposition with rainfall, dry deposition of dust containing N compounds, and gaseous deposition.

Samples were taken from selected environmental compartments such as biological crusts, sand underneath these crusts (down to a depth of 90?cm), N₂-fixing and non-N₂-fixing plants, atmospheric bulk deposition as well as soil from arable land north of the sandy area in three field campaigns in March 1998, 1999 and 2000. The δ¹⁵N values measured were in the following ranges: grass ?2.5‰ to +1.5‰; R. reatam: +0.5‰ to +4.5‰; non-N₂-fixing shrubs +1‰ to +7‰; sand beneath the biological crusts +4‰ to +20‰ (soil depth 2–90?cm); and arable land to the north up to 10‰. Thus, the natural ¹⁵N abundance of the different N pools varies significantly. Accordingly, it should be feasible to assess different input pathways from the various ¹⁵N abundances of nitrogen. For example, the biological N fixation rates of the Fabaceae shrub R. reatam from the ¹⁵N abundances measured were calculated to be 46–86% of biomass N derived from the atmosphere. The biological crusts themselves generally show slight negative ¹⁵N values (?3‰ to ?0.5‰), which can be explained by biological N fixation. However, areas with a high share of lichens, which are unable to fix atmospheric nitrogen, show very negative values down to ?10‰. The atmospheric N bulk deposition, which amounts to 1.9–3.8?kg?N/ha?yr, has a ¹⁵N abundance between 4.4‰ and 11.6‰ and is likely to be caused by dust from the arable land to the north. Thus, it cannot be responsible for the very negative values of lichens measured either. There must be an additional N input from the atmosphere with negative δ¹⁵N values, e.g. gaseous N forms (NO _x , NH₃). To explain these conflicting findings, detailed information is still needed on the wet, particulate and gaseous atmospheric deposition of nitrogen.     

Determining the content and 13C abundance of total dissolved carbon in water samples by TOC analyser-mass spectrometer coupling

A combined system consisting of a TOC analyser connected to a quadrupole MS was recently described as a way of measuring the N content and the 15N abundance of total dissolved nitrogen in aqueous samples. This work examines whether this combination of instruments can also be used for the 13C determination of the total dissolved carbon in aqueous samples. A level of precision good for 13C-enriched samples was achieved with a relative standard deviation of <3%. By using an isotope ratio MS instead of the quadrupole MS employed here, TOC-MS coupling also ought to be suitable for determining natural 13C abundances.     

Rapid, Sensitive and Highly Selective (15)N Analysis of (15)N Enriched Nitrite in Water Samples and Soil Extracts by Nitric Oxide Production and CF-QMS Measurement

Abstract Nitrite is a very important intermediate in many microbiological N transformations in soils and water. The stable isotope (15)N is often used to investigate these processes. The determination of (15)N in low concentrations of nitrite in the presence of large concentrations of nitrate is very difficult. Methods used so far for the isotope analysis of nitrite are unsatisfactory, because the nitrite must be calculated as the difference between nitrate plus nitrite and nitrate alone. More useful are mehods by which the nitrite is selectively converted into a chemical form that is suitable for (15)N analysis and that is free from interference from other N species, particularly nitrate. Using this principle in the present study we developed a method where the nitrite is reduced to nitric oxide by iodide in acid medium. This reaction is fast and quantitative, and the (15)N abundance of NO can be precisely measured by continuous flow mass spectrometry. This method is used for samples from tracer experiments with artificially enriched nitrogen 15. Therefore, the use of simple quadrupole mass spectrometers directly linked to the reaction unit is possible with sufficient precision (Reaction-Continuous Flow Quadrupole Mass Spektrometry-RCFQMS). Using the technique developed sample volumes up to 10ml containing at least 1.0 μg nitrite-N (0, 1 μg/ml) with a (15)N abundance of ? 0.42 at.% gave a precision of RSD ? ± 3%.     

Use of GC-QMS for stable isotope analysis of environmentally relevant main and trace gases in the air

Abstract

When tracing the fluxes of N compounds (especially N oxides) and carbon dioxide in the system soil-atmosphere and applying ¹⁵N and ¹³C, it is necessary to simultaneously determine both the concentration of the substances and their isotopic composition. GC separation (packed column) was coupled to a mass spectrometer by a special interface. Variable sample volumes can be delivered by changing the split by means of the interface in front of the MS. Thus, both the concentration of a gaseous compound and its ¹⁵N content can be measured. Supplying sample volumes in a range between 5 μl and 5 ml, it is possible to measure both components of high content (e.g., nitrogen) and at the lower ppm(v)-content (e.g., dinitrogen oxide) in the air. Measurements of N₂, N₂O, NO and CO₂ are demonstrated, too.     

Accuracy and Precision for Measurements of the Mass Ratio 30/28 in Dinitrogen from Air Samples and its Application to the Investigation of N Losses from Soil by Denitrification

Abstract

In the 1950s Hauck introduced a special version of the ¹⁵N dilution technique (¹⁵N flux method) for the determination of N losses from the soil by denitrification. Although this method is very useful and reliable its application has been rather infrequent up to now. This is mainly due to the need to measure the m/z 30 in addition to the usually measured m/z 28 and 29 for dinitrogen, because the ¹⁵N in the enriched air sample taken from an enclosure (cover box) at the soil surface is nonrandom. The signal from the m/z 30 is very low and difficult to measure with sufficient precision because other species (e.g. NO) also having the m/z 30 often interfere with its measurement. In this study the accuracy and precision of an easy to use CF-IRMS with sample batch operation to measure the ratio 30/28 was investigated. The relative standard deviation (RSD = precision) from natural abundance up to 2 at.% was always <1%. After correction of the mass ratio 30/28 (R30), by means of a formula obtained by linear regression of theoretical R30 against measured R30, the accuracy of the abundance calculated from this corrected R30 was very high. From the achieved precision and assuming a cover box height of 10 cm (headspace volume of 7 1), and a collection time of 2 h, a limit of detection for N₂ losses by denitrification equivalent to 16 g N/ha*d or 6 kg N/ha*a can be estimated. The performance of the ¹⁵N dilution method using the equipment and procedure described is demonstrated by means of results from an incubation experiment with [¹⁵N]nitrate-amended soils.     

15N, 13C-On-line Measurements with an Elemental Analyser (Carlo Erba,NA 1500), a Modified Trapping Box and a Gas Isotope Mass Spectrometer (Finnigan,MAT 251)

Abstract

An on-line method for the determination of ¹⁵N and ¹³C with a gas isotope mass spectrometer (Finnigan, MAT 251) was developed to improve the sensitivity and to reduce measurements time and the cost of the sample analysis. For this purpose an elemental analyser (Carlo Erba, NA 1500) was coupled to the mass spectrometer using parts of the capillary system of a trapping box (Finnigan, type CN). For the determination of samples with natural concentrations of ¹⁵N and ¹³C the uncertainty of the delta value is less than 0.2 δ‰. The detection limit is in the order of 10 μg (total N or total C) and 7 samples can be analysed per hour.     

Elemental Analyzer-Quadrupole MS Coupling—A Low Cost Equipment for the Simultaneous Determination of Carbon/13C and Nitrogen/15N in Isotopically Enriched Soil and Plant Samples

Abstract

Since the end of the 80s elemental analyzer-isotope ratio mass spectrometer connections have been used for the fast, automatic and highly precise determination of carbon and nitrogen content as well as their isotopic composition in one run. But for artificially enriched stable isotopes as tracer in biological processes and since these processes have a high biological variability anyway (e.g. soil processes) the use of these highly precise but also sophisticated and expensive instruments is not required. In this case the use of a quadrupole mass spectrometer connected with an elemental analyzer can offer a low cost alternative. As shown, such coupling is suitable for automatic simultaneous routine analysis of total nitrogen and carbon and their isotopic enrichment (¹⁵N, ¹³C) in plant material and soils. The relative standard deviation for ¹⁵N and ¹³C determination is 2% To meet this precision a careful sample homogenization by grinding is very important. The duration of one measurement is 6–8 min. depending on whether nitrogen alone or both nitrogen and carbon are determined. This enables a high sample throughput.     

Conference Reports

Abstract

One purpose of new land use concepts for degraded fens (organic soils with high N content) is the reduction of the mineralization process due to very high groundwater levels. However, knowledge of nitrogen mineralization process (net and gross) in degraded fen soils affected by reflooding is very small. Therefore, the objectives of our study were (a) to evaluate the suitability of ¹⁵N pool dilution method for measurements of gross mineralization rates in degraded fen soils and (b) to investigate how the reflooding of a degraded fen affects the net and gross nitrogen mineralization in a short-term incubation experiment. The usability of the ¹⁵N pool dilution method was diminished by the low recovery of the applied ¹⁵NH₄ ^? at time zero. The recovery of the added ¹⁵NH₄ ^? in the extractable soil NH₄ ^? pool was only 13.5% for the drained soil and 59.6% for the reflooded soil. However, the gross mineralization rates were similar for both soils and exceeded always the net rates substantially. The cumulative net mineralization rate was higher for the reflooded soil (1.58 μg N?cm^?3?d^?1) than for the drained soil (-0.67 μg N?cm^?3?d^?1). Differences between the two soils were also found in the nitrification intensity and the loss of ¹⁵N. This was probably one reason for the higher net mineralization rate in the reflooded soil.     

Automated and rapid online determination of 15N abundance and concentration of ammonium, nitrite, or nitrate in aqueous samples by the SPINMAS technique

On the basis of the principle of reaction continuous-flow quadrupole mass spectrometry, an automated sample preparation unit for inorganic nitrogen (SPIN) species was developed and coupled to a quadrupole Mass Spectrometer (MAS). The SPINMAS technique was designed for an automated, sensitive, and rapid determination of 15N abundance and concentration of a wide variety of N-species involved in nitrogen cycling (e.g. NH4+, NO3-, NH2OH etc.). In this paper, the SPINMAS technique is evaluated with regard to the determination of 15N abundance and concentration of the most fundamental inorganic nitrogen compounds in ecosystems such as NH4+, NO2-, and NO3-. The presented paper describes the newly developed system in detail and demonstrates the general applicability of the system. For a precise determination of 15N abundance and concentration, a minimum total N-amount of 10 microg NH4+ - N, 0.03 microg NO2- - N, or 0.3 microg NO3- - N has to be supplied. Currently, the SPINMAS technique represents the most rapid and only fully automated all-round method for a simultaneous determination of 15N abundance and total N-amount of NH4+, NO2-, or NO3- in aqueous samples.     

The use of stable isotopes in ecosystem research. First results of a field study with 15N

Terrestrial ecosystems, e.g. forest ecosystems, are characterized by a complex and sensitive network of biotic and abiotic factors and their interactions. By using stable isotopes (e.g. labelled nitrogen compounds), very small addition rates of highly enriched compounds can be applied, which do not change or disturb the investigated system, but provide information about single processes, their interactions and especially about their dynamics.

First results of a field study in the Fichtelgebirge, Northeast-Bavaria, Germany, are presented. The distribution of labelled nitrogen (as ¹⁵N-NH₄ ⁺ and ¹⁵N[sbnd]NO₃ ^?) within a spruce ecosystem (Picea abies (L.) Karst. in competition with understory vegetation of Vaccinium myrtillus, Calluna vulgaris and Deschampsia flexuosa) showed maximum ¹⁵N concentrations in tissues of the understory vegetation. During the first six weeks after the ¹⁵N application, the nitrogen uptake of all investigated species was higher after the ¹⁵N[sbnd]NO₃ ^? treatment than after the ¹⁵N[sbnd]NH₄ ⁺ treatment.     

Analysis of the coexisting pathways for NO and N2O formation in Chernozem using the 15N-tracer SimKIM-Advanced model

The nitrogen (N) cycle consists of a variety of microbial processes. These processes often occur simultaneously in soils, but respond differently to local environmental conditions due to process-specific biochemical restrictions (e.g. oxygen levels). Hence, soil nitrogen cycling (e.g. soil N gas production through nitrification and denitrification) is individually affected through these processes, resulting in the complex and highly dynamic behaviour of total soil N turnover. The development and application of methods that facilitate the quantification of individual contributions of coexisting processes is a fundamental prerequisite for (i) understanding the dynamics of soil N turnover and (ii) implementing these processes in ecosystem models. To explain the unexpected results of the triplet tracer experiment (TTE) of Russow et al. (Role of nitrite and nitric oxide in the processes of nitrification and denitrification in soil: results from ¹⁵N tracer experiments. Soil Biol Biochem. 2009;41:785–795) the existing SimKIM model was extended to the SimKIM-Advanced model through the addition of three separate nitrite subpools associated with ammonia oxidation, oxidation of organic nitrogen (N_org), and denitrification, respectively. For the TTE, individual treatments with ¹⁵N ammonium, ¹⁵N nitrate, and ¹⁵N nitrite were conducted under oxic, hypoxic, and anoxic conditions, respectively, to clarify the role of nitric oxide as a denitrification intermediate during N₂O formation. Using a split nitrite pool, this analysis model explains the observed differences in the ¹⁵N enrichments in nitric oxide (NO) and nitrous oxide (N₂O) which occurred in dependence on different oxygen concentrations. The change from oxic over hypoxic to anoxic conditions only marginally increased the NO and N₂O release rates (1.3-fold). The analysis using the model revealed that, under oxic and hypoxic conditions, N_org-based N₂O production was the dominant pathway, contributing to 90 and 50 % of the total soil N₂O release. Under anoxic conditions, denitrification was the dominant process for soil N₂O release. The relative contribution of N_org to the total soil NO release was small. Ammonia oxidation served as the major pathway of soil NO release under oxic and hypoxic conditions, while denitrification was dominant under anoxic conditions. The model parameters for soil with moderate soil organic matter (SOM) content were not scalable to an additional data set for soil with higher SOM content, indicating a strong influence of SOM content on microbial N turnover. Thus, parameter estimation had to be re-calculated for these conditions, highlighting the necessity of individual soil-dependent parameter estimations.     

Structure and quadrupole coupling measurements on the NO dimer

Rotational transition frequencies for ¹⁴NO-¹⁴NO, ¹⁴NO-¹⁵NO, and ¹⁵NO-¹⁵NO were measured using a pulsed-nozzle Fourier transform microwave spectrometer. Rotational constants for the different isotopic combinations allowed an unambiguous structure determination. The molecule is in a cis planar structure with a bond between the nitrogen atoms and an NNO angle θ = 99.6(2)°. The NN bond length is 2.236(1) Å and the NO bond length is 1.161(4) Å. Hyperfine structure due to nitrogen quadrupole coupling and spin-rotation interactions was observed and analyzed. Rotation constants, quadrupole coupling tensor, and spin-rotation tensor elements are given.     

Maternal obesity does not influence human milk protein 15N natural isotope abundance

ABSTRACT

Obesity increases protein metabolism with a potential effect on nitrogen isotope fractionation. The aim of this study was to test the influence of obesity on human milk extracted protein ¹⁵N natural isotope abundance (NIA) at one month post-partum and to compare human milk extracted protein ¹⁵N NIA and bulk infant hair ¹⁵N NIA. This cross-sectional observational study involved 16 obese mothers (body mass index (BMI)?≥?30?kg?m^?2 before pregnancy) matched with 16 normal-weight mothers (18.5?kg?m^?2?≤?BMI?<?25?kg?m^?2) for age and pregnancy characteristics. Human milk extracted protein and bulk infant hair ¹⁵N NIA were determined by isotope ratio monitoring by mass spectrometry interfaced to an elemental analyser (IRM-EA/MS). No significant difference was found in human milk protein ¹⁵N NIA values between obese and normal-weight mothers (8.93?±?0.48?‰ vs. 8.95?±?0.27?‰). However, human milk protein ¹⁵N NIA was significantly lower than bulk infant hair ¹⁵N NIA: 8.94?±?0.38?‰ vs. 9.66?±?0.69?‰, respectively. On the basis of these results, it is concluded that human milk protein ¹⁵N NIA measured at one month post-partum is not influenced by maternal obesity. These findings suggest that ¹⁵N NIA may be exploited to study metabolism without considering maternal obesity as a confounder.     

Airborne Nitrogen Input at Four Locations in the German State of Saxony-Anhalt - Measurements using the 15N-Based ITNI-System

Abstract

The amount of atmospheric N deposition in Germany is actual rather uncertain. Estimates using standard methods indicate an N deposition of 30–35 kg N/ha × year. However, the results of long-term field experiments and newly by the ITNI (Integrated Total Nitrogen Input) system could prove a much higher N input of about 50–60 kg N/ha × year. The reason for this difference is that standard methods use wet-only or bulk collectors, which neglect gaseous and organic N deposition as well as direct N uptake by aerial plant parts. By contrast, the ITNI-system is able to measure the total atmospheric N input using the ¹⁵N isotope dilution method. The input of airborne N into a soil/plant system leads to a dilution of the abundance of a previously applied ¹⁵N tracer over a defined time period. The atmospheric N deposition can be calculated from this dilution.

To estimate the actual N input in Central Germany, ITNI measurements were carried out from autumn 1998 to autumn 2000 at four locations in the German state of Saxony-Anhalt. Atmospheric N depositions between 45 and 75 kg N/ha × year were determined depending on the location. These results closely match to N balances of longterm field experiments. Furthermore, a relationship was found between N deposition and the plant species used as well as plant development.     

Synthese und Qualitätskontrolle 15N-markierter Stickoxide mit hoher 15N-Häufigkeit für umweltrelevante Tracer-Untersuchungen

Für die Untersuchung ausgewählter Probleme des Verhaltens und der Wirkung der Stickoxide NO _x (NO + NO₂) in Ökosystemen, z.B. die Aufnahme und Freisetzung von NOx durch das System “Boden-Pflanze”, bietet sich der Einsatz ¹⁵N-markierter Stickoxide an. Die dazu benötigten ¹⁵N-markierten Gasgemische hoher Reinheit werden aus eigens dafür synthetisiertem [¹⁵N]Stickstoffmonoxid oder [¹⁵N]Stickstoffdioxid mit hoher ¹⁵N-Häufigkeit hergestellt. Beide Synthesen gehen jeweils von der kostengünstig kommerziell erhältlichen [¹⁵N]Salpetersäure aus.

Im Falle des [¹⁵N]Stickstoffdioxids erfolgt die Herstellung über die Präparation von Bleinitrat und dessen thermische Zersetzung. Die Ausbeute liegt bei 70–75% bezogen auf eingesetzte [¹⁵N]Salpetersäure.

Die Herstellung von [¹⁵N]Stickstoffmonoxid erfolgt durch Reduktion von [¹⁵N]Salpetersäure mit Eisen-II-sulfat in stark saurer Lösung. Die Ausbeute beträgt 60–70%, bezogen auf eingesetzte Salpetersäure.

The application of ¹⁵N is very useful for the investigation of the behavior and the effect of the nitrogen oxides NO _x (nitric oxide + nitrogen dioxide) in ecosystems, e.g. the uptake and release of NO _x by the soil-plant system. The ¹⁵N labelled gas mixture needed for that purpose has to be prepared from synthesized highly enriched [¹⁵N]nitric oxide and [¹⁵N]nitrogen dioxide. These two syntheses both use the commercially available and reasonable [¹⁵N]nitric acid.

In the case of [¹⁵N]nitrogen dioxide the synthesis is carried out via [¹⁵N]lead nitrate and its decomposition with increasing temperature. The yield is 70–75% related to the [¹⁵N]nitric acid input. The preparation of [¹⁵N]nitric oxide is done by reduction of [¹⁵N]nitric acid by means of FeSO₄ in strong acid solution. The yield amounts to 60–70%.     

Time resolved simultaneous detection of 14NO and 15NO via mid-infrared cavity leak-out spectroscopy

We present a ring-down absorption spectrometer based on a continuous-wave CO laser in the mid-infrared spectral region near λ?=?5 μm. Using a linear ring-down cavity (length: 0.5 m) with high reflective mirrors (R?=?99.988 %), we observed a noise-equivalent absorption coefficient of 3?×?10^?10 cm^?1Hz^?1/2. This corresponds to a noise-equivalent concentration of 800 parts per trillion (ppt) for ¹⁴NO and 40 ppt for ¹⁵NO in 1 s averaging time. We achieve a time resolution of 1 s which allows time resolved simultaneous detection of the two N isotopes. The δ¹⁵N value was obtained with a precision of ±1.2‰ in a sample with a NO fraction of 11 ppm. The simultaneous detection enables the use of ¹⁵NO as a tracer molecule for endogenous biomedical processes.