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Seawater samples for analysis of the N isotopic composition of nitrate+nitrite and nitrate-only were collected unfiltered at regular depth intervals from the surface to 1000 m in 60 ml (>150 m) or 125 ml (<150 m) square-bottomed, wide-mouth HDPE bottles (Nalgene). Bottles were acid-washed and rinsed with deionized water prior to sampling. At sea, pre-labelled bottles and caps were rinsed three times with sample water, filled to ~85% of the bottle volume, and frozen upright at -20\u00b0C until analysis.<\/p>\n
Isotopic analyses were conducted using the \u201cdenitrifier method\u201d, wherein denitrifying bacteria lacking nitrous oxide (N2O) reductase quantitatively convert nitrate and nitrite in the sample to N2O gas (Sigman et al. 2001, Casciotti et al. 2002) (see also (Weigand et al. in review) for the updated protocol used for analyzing these samples). The isotopic composition of N2O was then measured by gas chromatography-isotope ratio mass spectrometry (GC-IRMS) using a purpose-built on-line N2O extraction and purification system and a Thermo MAT 253 mass spectrometer. Seawater solutions of the international nitrate reference materials, IAEA-N3 and USGS34, as well as an in-house N2O standard, were run in parallel to the samples in order to monitor the quality of bacterial N conversion and mass spectrometric measurements. The reference materials bracketed each group of ~10 samples and were used to correct the measured \u03b415N to N2 in air (Sigman et al. 2001, Casciotti et al. 2002, McIlvin & Casciotti 2011). \u00a0\u00a0\u00a0\u00a0\u00a0\u00a0<\/p>\n
The measurement of the 15N of nitrate-only for samples with a detectable concentration of nitrite required a nitrite removal pre-treatment. The detection limit for nitrite in this case was 2 nmol kg-1. Samples collected between the surface and ~125 m were treated for nitrite removal via the addition of 10 \u00b5l of sulphamic acid solution per ml of sample, which converts sample nitrite to N2 gas with a reaction time of 2-8 minutes, followed by the addition of 5.5 \u00b5l of 2M NaOH per ml of sample to restore the pH of the sample to ~7-9 (Granger & Sigman 2009). The pooled standard error for 15N was 0.04\u2030 and 0.11\u2030 (n \u22653) for nitrate+nitrite and nitrate concentrations \u22650.5 \u00b5mol l-1 and <0.5 \u00b5mol l-1, respectively. Hereafter, \u201cnitrate\u201d in the text refers to nitrate-only, after the subtraction (for concentration) or removal (for 15N) of nitrite.<\/p>\n
Suspended particulate N: Suspended PN was collected at various depths throughout the euphotic zone, including within the surface mixed layer and at the depth of maximum chlorophyll concentration, by gentle vacuum filtration (<135 mbar), of 8 l of seawater through a GF-75 filter. Filters were transferred to pre-combusted (500\u00b0C for 5 h) aluminium foil envelopes, and immediately frozen at -80\u00b0C until analysis. In the laboratory, the PN filters were dried in a desiccating oven at 40\u00b0C. Three subsamples were cored from each filter and transferred to combusted 4 mL glass Wheaton vials. PN was oxidised to nitrate using the persulphate oxidation method of Knapp et al. (2005), and as modified by Fawcett et al. (2011; 2014); this was conducted in a laminar flow hood equipped with an ammonia\/amine filter. Briefly, 2 ml of persulphate oxidizing reagent (POR) were added to each sample vial, as well as to triplicate vials containing a filter blank plus varying quantities of two L-glutamic acid isotope standards, USGS-40 and USGS-41 (Qi et al. 2003); this allows determination of the N content and 15N of the POR+filter blank. The POR was made by dissolving 2.5 g of 4\u00d7 recrystallised, methanol-rinsed potassium persulphate and 2.5 g of sodium hydroxide in 100 ml of ultra high-purity deionised water. Following POR addition, vials were autoclaved at 121\u00b0C for 55 minutes on a slow-vent setting, after which sample pH was lowered to 5-8 using 12N HCl. The concentration and \u03b415N of the resultant nitrate was measured via chemiluminescent analysis (Braman & Hendrix 1989) and the denitrifier method (see above) (Sigman et al. 2001, Casciotti et al. 2002). The final N content and \u03b415N of the oxidised samples was corrected for the POR+filter blank. N content was converted to PN concentration by normalising to whole-filter area and volume of seawater filtered.<\/p>\n
References:<\/p>\n
Braman RS, Hendrix SA (1989) Nanogram nitrite and nitrate determination in environmental and biological materials by vanadium(iii) reduction with chemi-luminescence detection. Anal Chem 61:2715-2718<\/p>\n
Casciotti K, Sigman D, Hastings MG, B\u00f6hlke J, Hilkert A (2002) Measurement of the oxygen isotopic composition of nitrate in seawater and freshwater using the denitrifier method. Anal Chem 74:4905-4912<\/p>\n
Fawcett SE, Lomas M, Casey JR, Ward BB, Sigman DM (2011) Assimilation of upwelled nitrate by small eukaryotes in the Sargasso Sea. Nature Geoscience 4:717-722<\/p>\n
Fawcett SE, Lomas MW, Ward BB, Sigman DM (2014) The counterintuitive effect of summer\u2010to\u2010fall mixed layer deepening on eukaryotic new production in the Sargasso Sea. Glob Biogeochem Cycle 28:86-102<\/p>\n
Granger J, Sigman DM (2009) Removal of nitrite with sulfamic acid for nitrate N and O isotope analysis with the denitrifier method. Rapid Commun Mass Spectrom 23:3753-3762<\/p>\n
Knapp AN, Sigman DM, Lipschultz F (2005) N isotopic composition of dissolved organic nitrogen and nitrate at the Bermuda Atlantic Time\u2010series Study site. Glob Biogeochem Cycle 19<\/p>\n
McIlvin MR, Casciotti KL (2011) Technical updates to the bacterial method for nitrate isotopic analyses. Anal Chem 83:1850-1856<\/p>\n
Qi H, Coplen TB, Geilmann H, Brand WA, B\u00f6hlke J (2003) Two new organic reference materials for \u03b413C and \u03b415N measurements and a new value for the \u03b413C of NBS 22 oil. Rapid Commun Mass Spectrom 17:2483-2487<\/p>\n
Sigman D, Casciotti K, Andreani M, Barford C, Galanter M, B\u00f6hlke J (2001) A bacterial method for the nitrogen isotopic analysis of nitrate in seawater and freshwater. Anal Chem 73:4145-4153<\/p>\n
Weigand MA, Foriel J, Barnett B, Oleynik S, Sigman DM (in review) Updates to instrumentation and protocols for isotopic analysis of nitrate by the denitrifier method. Rapid Commun Mass Spectrom<\/p><\/div>","@type":"rdf:HTML"}],"http:\/\/ocean-data.org\/schema\/hasBriefDescription":[{"@value":"Particulate nitrogen concentrations and N isotopic composition, and nitrate isotopic composition from EN532","@language":"en-US"}],"http:\/\/purl.org\/dc\/terms\/description":[{"@value":"
Pico- and nanoplankton cell concentrations\u00a0from\u00a0CTD casts made during the August-September 2013 EN532 and April-May 2014 EN358 cruises aboard R\/V Endeavor. Study sites in the subarctic Atlantic Ocean along the 20 \u00b0W meridian between 50 \u00b0N and 60 \u00b0N in September 2013 and May 2014. Two transects from the US East coast to the subarctic study sites were performed as well.<\/p>\n
Related Dataset:<\/strong> Standard deviations are derived from at least two analytical measurements<\/p>\n BCO-DMO Processing:<\/strong><\/p>\n -\u00a0added conventional header with dataset name, PI name, version date - replaced\u00a0original 2016-07-13 EN532 data with new version submitted 2017-07-14. 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EN532 - CTD<\/a>
EN538 - CTD<\/a>
Chlorophyll-a: EN532 and EN538<\/a>
Nutrients: EN532 and EN538<\/a>
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