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Data was acquired from an isotope ratio mass spectrometer using Isodat 3.0 software.<\/p>\n
Dinitrogen (N2) calculations:<\/strong> Calculation: Constants: The following equations were applied to air standards to correct for the blank 30\/28: The blank is then removed from the measured 30A\/28A ratio of samples: Added 30N2 is calculated: Total 15N added = 2 *\u00a0 30N2added + 29N2added. However, 29N2added was negligible, as expected from the 98+ atom % 15N2 gas. Therefore, the reported 15N2 gas added is equivalent to added 30N2.<\/p>\n Summary of methods from Dabundo et al. 2014:<\/strong> Preparation of nitrate & ammonium solutions:<\/em> Initial experiments consisted of 40 mL or 100 mL solutions of 10, 50, 100, 200, or 300 umol\/L nitrate and 5 umol\/L ammonium chloride in 60 mL or 120 mL serum vials that were sealed with stoppers. The 20 mL of air headspace in each of the treatment vials was supplemented with 0.1 mL of 15N2 gas from respective bottles from each of the three suppliers. The solutions were equilibrated overnight on a shaker, after which the 15N\/14N and 18O\/16O isotope ratios of nitrate were analyzed. The 15N\/14N isotope ratio of ammonium was also analyzed in experimental solutions treated with the Sigma-Aldrich and Cambridge Isotopes stocks.<\/p>\n Additional experiments were carried out in which 2 mL 15N2 gas was equilibrated overnight in 20 mL serum vials containing 10 mL solutions of 10 umol\/L sodium nitrate, after which the 15N\/14N and 18O\/16O ratios of nitrate were measured. Similarly, 10 mL solutions of 5 umol\/L ammonium chloride were dispensed in 20 mL serum vials and equilibrated overnight with 2 mL 15N2 gas, after which the 15N\/14N isotope ratios of ammonium were analyzed.<\/p>\n Nitrate and ammonium concentrations:<\/em> Nitrate N and O isotope ratio analyses:<\/em> The 15N\/14N reference is N2 in air, and the 18O\/16O reference is Vienna Standard Mean Ocean water (V-SMOW). Individual analyses on the GC-IRMS were referenced to injections of N2O from a pure N2O gas cylinder, and then standardized through comparison to the international nitrate standards USGS-34, USGS-32, and IAEA-NO-3, using standard bracketing techniques. Nitrate samples from experiments with Campro Scientific 15N2 were standardized with USGS-32 and IAEA-NO-3, and an additional internal lab nitrate standard (UBN-1).<\/p>\n Headspace N2 isotope ratio analyses:<\/em> The measured quantity of 15N2 added to nitrate and ammonium solutions equilibrated with 15N2 gas.<\/p>\n Refer to the following publication for more information: See related datasets: Samples with the same ID are replicated measurements.<\/p>\n BCO-DMO Edits:
\nMoles of 15N2 added were calculated from 28, 29, and 30 areas compared to the atmosphere.<\/p>\n
\natm = atmosphere
\nvariables labeled \u201cA\u201d correspond to measured area values<\/p>\n
\n15Natm mole fraction = 0.003663
\n14Natm mole fraction = 0.996337
\nTherefore the fraction of\u00a0 30N2atm = 1.34176E-05; 29N2atm = 0.007299165; 28N2atm = 0.992687418
\n30N2atm\/ 28N2atm = 1.35164E-05 = 30\/28natural_abundance
\n28N2 total =\u00a0 28N2 total in incubation was calculated from the ideal gas law at 25 degrees C and Henry\u2019s Law. Found to be 654 umol in incubations with 40 mL solution and 20 mL headspace, and 690 umol in incubations with 100 mL solution and 20 mL headspace.<\/p>\n
\n30area from binomial = 28A * 30\/28nat_abundance
\n30\/28area ratio blank = (30A\/28A) \u2013 30area_from_binomial\/28A
\n30\/28area ratio blank is graphed (y axis) vs. 28A (x axis) to account for variation relating to the injection volume.
\nThe slope (mratio blank) and intercept (iratio blank) of the corresponding linear regression are found.<\/p>\n
\n30A\/28A real = 30A\/28A \u2013 (mratio_blank * 28A + iratio_blank)<\/p>\n
\n30N2total = 30A\/28A real * 28N2total
\n30N2added = 30N2total \u2013 (28N2total * 30\/28natural_abundance)<\/p>\n
Reagents:<\/em>
\nFour lecture bottles of 98+ at% 15N-labeled N2 gas were purchased from Sigma-Aldrich, three from lot # SZ1670V, and one from lot # MBBB0968V. Two 1L lecture bottles of 98+ at% 15N2 were purchased from Cambridge Isotopes from lot #\u2019s I1-11785A and I-16727. One 1L lecture bottle of 98+ at% 15N2 was purchased from Campro Scientific from lot # EB1169V. Ammonium and nitrate solutions were prepared with salts or with solutions obtained from different distributors: sodium nitrate (NaNO3), potassium nitrate (KNO3), and ammonium chloride (NH4Cl) from Fisher Scientific; analytical-grade potassium nitrate from Fluka Analytical and a gravimetric solution of ammonium chloride from SPEX CertiPrep.<\/p>\n
\nAqueous solutions of natural abundance (unlabeled) ammonium and nitrate salts were equilibrated overnight with an air headspace supplemented with an injection of 15N2 gas (to determine whether the 15N2 gas stocks contained 15N-labeled ammonia (NH3) or nitrate and\/or nitrite (NOx) contaminants). After equilibration, the 15N\/14N ratio of ammonium and the 15N\/14N and 18O\/16O ratios of nitrate\/nitrite in solution were measured, as well as the 15N\/14N ratio of N2 gas in the headspace. The isotope ratios of nitrate and ammonium were compared to those in control solutions, which were not supplemented with 15N2 gas. Experiments with the Campro Scientific 15N2 stock were verified for 15N-nitrate\/nitrite contaminants only (and not for 15N-ammonium).<\/p>\n
\nNitrate concentrations in the experimental solutions were verified via reduction to nitric oxide in hot vanadium (III) solution followed by detection with a chemiluminescence NOx analyzer (model T200 Teledyne Advanced Pollution Instrumentation). Ammonium concentrations were measured by derivatization with orthophthaldialdehyde (OPA) and fluorometric detection on an AJN Scientific f-2500 Fluorescence Spectrophotometer.<\/p>\n
\nNitrate\/nitrite nitrogen (15N\/14N) and oxygen (18O\/16O) isotope ratios were measured using the denitrifier method. Nitrate (and nitrite) in experimental samples was converted stoichiometrically to nitrous oxide (N2O) by a denitrifying bacterial strain (Pseudomonas chlororaphis f. sp. aureofaciens, ATCC 13985) that lacks nitrous oxide reductase. The N and O isotopic composition of N2O was then measured on a Delta V Advantage Isotope Ratio Mass Spectrometer (IRMS) interfaced with a modified Gas Bench II gas chromatograph (Thermo Fisher) purge and trap system. The isotope ratio measurements are reported in per mille (o\/oo) units.<\/p>\n
\nTo measure the d15N of N2 gas in the headspace of experimental samples, 75 uL of headspace was injected into 12 mL Exetainer vials previously flushed with helium, then analyzed on a Gas Bench II GC-IRMS (Delta V Advantage Plus) operated in continuous flow mode. N2 and (O2+ Ar) were separated on a gas chromatography column. The analyses were standardized with parallel analyses of ambient N2 gas in air. These direct N2 gas measurements were carried out for experiments conducted using two of three lecture bottles from Sigma-Aldrich lot, and for experiments conducted using the lecture bottle from Cambridge Isotopes. The 15N2 concentration in the headspace of other experiments was estimated from the tracer injection volume rather than from direct measurements.<\/p><\/div>","@type":"rdf:HTML"}],"http:\/\/ocean-data.org\/schema\/hasBriefDescription":[{"@value":"The measured quantity of 15N2 added to nitrate and ammonium solutions equilibrated with 15N2 gas.","@language":"en-US"}],"http:\/\/purl.org\/dc\/terms\/description":[{"@value":"
\nDabundo, R., Lehmann, M.F., Treibergs, L., Tobias, C.R., Altabet, M.A., Moisander, P.H., and Granger, J. 2014. The Contamination of Commercial 15N2 Gas Stocks with 15N\u2013Labeled Nitrate and Ammonium and Consequences for Nitrogen Fixation Measurements. PLoS ONE, 9(10): e110335. doi:10.1371\/journal.pone.0110335<\/a><\/p>\n
delta 15N NO3<\/a>
delta 15N NH4<\/a>
direct N2O<\/a>
particulate N<\/a><\/p><\/div>","@type":"rdf:HTML"}],"http:\/\/www.w3.org\/2000\/01\/rdf-schema#label":[{"@value":"N2","@type":"xsd:string"}],"http:\/\/ocean-data.org\/schema\/hasProcessingDescription":[{"@value":"
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