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Hydrolysis, derivatization and analyses of samples for CSIA \u2013\u00a0 For Exps. 3-4, samples (1-2 mg dry weight) for CSIA-AA were added to 1 ml 6 N HPLC-grade HCl, then flushed with N2, capped with a Teflon-lined cap, and hydrolyzed at 150\u00b0C for 70 min.\u00a0 The resulting hydrolysate was evaporated to dryness under N2 at 55\u00b0C, redissolved in 1 ml 0.01 N HCl, purified by filtration (0.45-\u03bcm hydrophilic filter), washed with 1 ml of 0.01 N HCl, and further purified using cation-exchange chromatography with a 5-cm resin column (Dowex 50WX8-400) in a glass Pasteur pipette.\u00a0 AAs were eluted with 4 ml of 2 N NH4OH and evaporated to dryness under a stream of N2 at 80\u00b0C, then reacidified with 0.5 ml of 0.2 N HCl, flushed with N2, heated to 110\u00b0C for 5 min and evaporated to dryness under N2 at 55\u00b0C.\u00a0 Hydrolyzed samples were esterified with 2 ml of 4:1 isopropanol:acetyl chloride, flushed with N2 and heated to 110\u00b0C for 60 min.\u00a0 After drying at 60\u00b0C under N2, the samples were acylated by adding 1 ml of 3:1 methylene chloride:trifluoracetic anhydride (TFAA) and heated to 100\u00b0C for 15 min.\u00a0 The derivatized AAs were further purified by solvent extraction following (Ueda et al. 1989).\u00a0 The acylated AA esters were evaporated at room temperature under N2 and redissolved in 3 ml of 1:2 chloroform:P-buffer (KH2PO4 + Na2HPO4 in Milli-Q water, pH 7).\u00a0 Vigorous shaking ensured that the derivitized AAs were partitioned into chloroform while contaminants remained in the P-buffer.\u00a0 The solvents were separated by centrifugation (10 min at 600 g), the chloroform was transferred to a clean vial, and the solvent extraction process repeated.\u00a0 Finally, to ensure derivatization, the acylation step was repeated.\u00a0 Samples were stored at \u201320\u00b0C in 3:1 methylene chloride:TFAA for up to 2 weeks until analysis.\u00a0 AA derivatives were then analyzed by isotope monitoring gas chromatography-mass spectrometry.\u00a0 We used a Delta V Plus mass spectrometer interfaced with a Trace GC gas chromatograph through a GC-C III combustion furnace (980\u00b0C), reduction furnace (650\u00b0C), and liquid nitrogen cold trap.\u00a0 Internal reference compounds (aminoadipic acid and norleucine) of known nitrogen isotopic composition were co-injected with samples and used to normalize the measured \u03b415N values of unknown AAs, and a suite of eight AAs with known isotopic composition was analyzed every 3 injections for additional quality control.\u00a0 At least three injections per sample were analyzed.\u00a0 For Exps. 5 and 6, CSIA-AA samples were hydrolyzed in 6N HCl at 110\u00b0C for 20 h.\u00a0 The hydrolysate was evaporated to dryness at temperature under a continuous stream of N2 gas, then esterified with 4:1 isotopropanol:acetyl chloride at 110\u00b0C for 60 min, acylated in a 1:1 solution of methylene chloride:trifluoracetic anhydride (DCM:TFAA) for 10 min at 110\u00b0C.\u00a0 Samples were stored at -20\u00b0C in 1:1 DCM:TFAA for up to 3 months before isotope analysis using a Delta V Plus mass spectrometer (Thermo Scientific) interfaced through a Conflo IV to a GC 1310 gas chromatograph coupled to a GC Isolink combustion-reduction furnace (1000\u00b0C) and liquid nitrogen cold trap.\u00a0 All samples were injected (splitless injector) onto a forte BPx5 capillary column (60 m x 0.32 mm x 1.0-\u03bcm film thickness) at an injector temperature of 250\u00b0C with a constant helium flow rate of 1.4 ml min\u20131.\u00a0 The column was initially held at 50\u00b0C for 2 min and then increased to 125 \u00b0C at a rate of 15\u00b0C min-1.\u00a0 Once at 125\u00b0C, the temperature was increased at a rate of 3\u00b0C min-1 to 160\u00b0C and then to 190\u00b0C at a rate of 4\u00b0C min-1.\u00a0 The final temperature of 300\u00b0C was reached by ramping to 275\u00b0C at 6\u00b0C min-1 and then 15\u00b0C min-1 afterward.\u00a0 Samples were analyzed in triplicate and normalized to the known \u03b415N values of a suite of 14 AAs analyzed before and after each set of 3 samples.<\/p><\/div>","@type":"rdf:HTML"}],"http:\/\/ocean-data.org\/schema\/hasBriefDescription":[{"@value":"15N AA from phytoplankton, microzooplankton and Calanus pacificus","@language":"en-US"}],"http:\/\/purl.org\/dc\/terms\/description":[{"@value":"
15N AA from phytoplankton, microzooplankton, and Calanus pacificus.<\/em><\/p><\/div>","@type":"rdf:HTML"}],"http:\/\/www.w3.org\/2000\/01\/rdf-schema#label":[{"@value":"Zooplankton\/phytoplankton CSIA-AA","@type":"xsd:string"}],"http:\/\/ocean-data.org\/schema\/hasProcessingDescription":[{"@value":" Data were processed using R software to correct sample values using a standard curve of known versus measured \u03b415N values.\u00a0 The standard deviations of measured \u03b415N values of standard AAs ranged from 0.1 to 1.7\u2030.\u00a0 Analyses of replicate samples are noted in data\u00a0table.\u00a0\u00a0<\/p>\n BCO-DMO Processing:<\/strong>
\n- modified parameter names to conform with BCO-DMO naming conventions;
\n- replaced spaces with underscores in organism and stage columns;
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