http://lod.bco-dmo.org/id/dataset/713831
eng; USA
utf8
dataset
Highest level of data collection, from a common set of sensors or instrumentation, usually within the same research project
Biological and Chemical Oceanography Data Management Office (BCO-DMO)
Unavailable
508-289-2009
WHOI MS#36
Woods Hole
MA
02543
USA
info@bco-dmo.org
http://www.bco-dmo.org
Monday - Friday 8:00am - 5:00pm
For questions regarding this resource, please contact BCO-DMO via the email address provided.
pointOfContact
2017-08-24
ISO 19115-2 Geographic Information - Metadata - Part 2: Extensions for Imagery and Gridded Data
ISO 19115-2:2009(E)
Amino acid isotopes from mussels along the California margin from 2009-2013 (Amino Acid Sediment 15N project)
2017-08-24
publication
2017-08-24
revision
Marine Biological Laboratory/Woods Hole Oceanographic Institution Library (MBLWHOI DLA)
2019-06-11
publication
https://doi.org/10.1575/1912/bco-dmo.713831.1
Matthew D. McCarthy
University of California-Santa Cruz
principalInvestigator
Biological and Chemical Oceanography Data Management Office (BCO-DMO)
Unavailable
508-289-2009
WHOI MS#36
Woods Hole
MA
02543
USA
info@bco-dmo.org
http://www.bco-dmo.org
Monday - Friday 8:00am - 5:00pm
For questions regarding this resource, please contact BCO-DMO via the email address provided.
publisher
Cite this dataset as: McCarthy, M. (2017) Amino acid isotopes from mussels along the California margin from 2009-2013 (Amino Acid Sediment 15N project). Biological and Chemical Oceanography Data Management Office (BCO-DMO). (Version 1) Version Date 2017-08-24 [if applicable, indicate subset used]. doi:10.1575/1912/bco-dmo.713831.1 [access date]
N, C compound-specific amino acid isotopes from mussels along CA margin: new approach for paleo-isoscape reconstructions Dataset Description: <p>N, C compound-specific amino acid&nbsp; isotopes from mussels along CA margin:&nbsp; new approach for paleo-isoscape reconstructions.</p> Methods and Sampling: <p><strong>Sampling</strong></p>
<p>Mussels were collected in the winter (Dec – Feb) of 2009–2010. Sites were chosen to be approximately evenly distributed along the CA coastline, with ,80 km geographic separation between each</p>
<p>sampling site. Our main goal here was to sample mussels from a wide geographic range across the CCS, although for observing finer scale local or regional variations, a finer-scale sampling strategy would like be required. Typically 5 individual mussels were collected from each site, all between 30–40 mm maximum shell length, which were immediately placed on dry ice until further preparation. The adductor muscle of each individual was dissected for analysis. This tissue was selected because isotopic values in muscle tissue have shown relatively long turnover times; based on past growth data, mussels of this size would be expected to integrate approximately annual variability in suspended food source isotopic values for each location sampled.</p>
<p>The dissected adductor tissue was carefully separated from other tissue types, rinsed with deionized water, refrozen, and then freeze-dried for 48 hrs. Lipids were removed &nbsp;using petroleum ether in a Dionex Accelerated Solvent Extractor (Bannockburn, IL). Finally, in preparation for CSI-AA, composite samples were made from a subset of 13 collection sites. For each location chosen for CSI-AA (based on the bulk d15N record), 160.05 mg of lyophilized tissue was weighed and combined for each individual mussel (n = 5).</p>
<p><strong>Elemental and Bulk Isotopic Analyses</strong></p>
<p>Stable carbon (d13C) and nitrogen (d15N) isotope ratios were determined via elemental analyzer isotope ratio mass spectrometry (EA-IRMS) at the University of California, Santa Cruz, Stable Isotope Laboratory (UCSC-SIL; http://emerald.ucsc.edu/~silab/). Approximately 1 mg of each dry isolated DOM sample was weighed into tin capsules (Costec, 5 x 9 mm) for analysis. EA-IRMS analysis was conducted using a Carlo Erba CHNS-O EA1108-elemental analyzer interfaced via a ConFlo III device with a ThermoFinnigan Delta Plus XP isotope ratio mass spectrometer (Thermo Fisher Scientific). &nbsp;Standards, EA-IRMS protocols, and correction routines followed standard UCSC-SIL protocols. Analytical uncertainties of n=3 replicate measurements of isotopic standards ranged from ± 0.05 to 0.1‰ for both d13C and d15N. Carbon to nitrogen elemental ratios were similarly determined by elemental analysis. The presented ratios are atomic ratios (C/N)a normalized to the mass of C and N, but have been abbreviated as C/N throughout.</p>
<p><strong>Compound-specific amino acid Isotopic 13C Analyses</strong></p>
<p>Individual AA d13C values were measured as trifluoroacetyl isopropyl ester (TFA-IP) AA derivatives, after acid hydrolysis. Samples were hydrolyzed by adding 2.5 mg homogenized composite muscle tissue to 1 ml of 6 N HCl, and heating for 20 h at 110 deg C under nitrogen. After drying, AA isopropyl esters were prepared with a 1:5 mixture of AcCl:2-propanol (110 deg C, 60 min) and then acylated using a 1:3 mixture of di chloro - methane (DCM) and trifluoroacetic anhydride (TFAA) (100 deg C, 15 min). Derivatized AAs were dissolved in DCM to a final ratio of approximately 2.5 mg of original tissue to 250 ul DCM for injection on the gas chromatograph IRMS (GC-IRMS) system.</p>
<p>Isotopic analysis was conducted on a Thermo Trace GC Ultra with inline oxidation and reduction furnaces, coupled to a ThermoFinnigan Delta Plus XP IRMS, equipped with a CTC Analytics autosampler.Derivatives (1 ul) were injected (250 deg C constant temperature) onto an Agilent DB-5 column (50 m × 0.32 mm ID × 0.52 um film thickness), with a He carrier flow rate of 2 ml min−1 (constant flow). Separations were achieved with a 4-ramp oven program: 52 deg C, 2 min hold; ramp 1 = 15 deg C min−1 to 75 deg C, hold for 2 min; ramp 2 = 4 deg C min−1 to 185 deg C, hold for 2 min; ramp 3 = 4 deg C min−1 to 200 deg C; ramp 4 = 30 deg C min−1 to 240 deg C, hold for 5 min.</p>
<p>This method allowed for d13C determination of the following AAs in mussel tissue: non-essential AAs alanine (Ala), aspartic acid + asparagine (Asp), glutamic acid + glutamine (Glu), glycine (Gly), proline (Pro), serine (Ser), and tyrosine Tyr); and essential AAs leucine (Leu), isoleucine (Ile), valine (Val), phenylalanine (Phe), and threonine (Thr). Acid hydrolysis destroys tryptophan and cystine, so these were not detected, and it also deaminates\ asparagine to aspartic acid, and glutamine to glutamic acid. While the abbreviations Glx and Asx\ are sometimes used to denote the combined Gln+Glu and Asn+Asp peaks, in order to correspond better with extant CSI-AA literature, we have elected to simply use Asp and Glu abbreviations, as defined above.</p>
<p>All samples were analyzed on the GC-IRMS in triplicate, and measured AA d13C values were corrected for the C added during derivatization, following the approach of Silfer et al. (1991). Reproducibility for tissue samples was typically less than &lt;0.3‰ (n = 3). The average mean deviation for all tissue sample replicates was 0.4‰.</p>
<p><strong>Compound-specific amino acid Isotopic 15N Analyses</strong></p>
<p>Amino acid d15N values were measured as Trifluoroacetyl isopropyl ester (TFA-IP) AA derivatives, following protocols described in detail elsewhere (e.g.,. Briefly, samples were hydrolyzed (6 N HCl, 20 hr at 110uC) under nitrogen, and TFA derivatives subsequently prepared from free AA using a modified version of the protocol described by Silfer (Silfer et al. 1991): isopropyl esters were made with a 1:5 mixture of Acetyl Chloride (AcCl):2-propanol (110uC, 60 minutes), and then acylated using a 1:3 mixture of Dichloromethane:Trifluroacetyl acetate (DCM:TFAA) (100uC, 15 minutes). Derivatized AAs were dissolved in DCM to a final ratio of approximately 4 mg of original tissue to 250 ml DCM. After derivatization, samples were analyzed by a Varian gaschromatograph coupled to a Finnegan Delta-Plus isotope ratiomass spectrometer (GC-IRMS). AAs were separated using a 50 m, 0.32 ID Hewlett Packard Ultra-1 column with 1 mm film thickness. Under our analytical conditions, d15N values could be reproducibly measured for alanine (Ala), aspartic acid + asparagine (Asp), glutamic acid + glutamine (Glu), leucine (Leu), isoleucine (Ile), proline (Pro), valine (Val), glycine (Gly), lysine (Lys), serine (Ser), phenylalanine (Phe), threonine (Thr), and tyrosine (Tyr) (Fig. S4). Most AAs were measured with a standard error of ,1.0% (based on n = 4 injections), and the average mean deviations for individual AA d15N measurements across all tissue sample replicates was 0.5%.</p>
Funding provided by NSF Division of Ocean Sciences (NSF OCE) Award Number: OCE-1131816 Award URL: https://www.nsf.gov/awardsearch/show-award?AWD_ID=1131816
completed
Matthew D. McCarthy
University of California-Santa Cruz
831-459-1533
1156 High Street
Santa Cruz
CA
95064
USA
mdmccar@ucsc.edu
pointOfContact
asNeeded
Dataset Version: 1
Unknown
Site
Identifier
Habitat_Type
n
Latitude_degMin
Longitude_degMin
lat
lon
Bulk_d15N
Bulk_d15N_stdev
Bulk_d13C
Bulk_d13C_stdev
Average_Trophic
Average_Source
Trophic_Position
Ala
Ala_stdev
Asx
Asx_stdev
Glx
Glx_stdev
Gly
Gly_stdev
Ile
Ile_stdev
Leu
Leu_stdev
Lys
Lys_stdev
Phe
Phe_stdev
Pro
Pro_stdev
Ser
Ser_stdev
Thr
Thr_stdev
Tyr
Tyr_stdev
Val
Val_stdev
element
Carlo Erba CHNS-O EA1108-elemental analyzer
Trace GC, interfaced via a ConFlo III and GCC device
hermoFinnigan Delta Plus XP isotope ratio mass spectrometer (Thermo Fisher Scientific)
theme
None, User defined
site
No BCO-DMO term
site description
count
latitude
longitude
d15N
d13C
alanine
amino acid concentration
glycine
isoleucine
leucine
lysine
phenylalanine
serine
threonine
tyrosine
valine
element
featureType
BCO-DMO Standard Parameters
Elemental Analyzer
Gas Chromatograph
Isotope-ratio Mass Spectrometer
instrument
BCO-DMO Standard Instruments
McCarthy_2009
service
Deployment Activity
California Coast Littoral Waters
place
Locations
otherRestrictions
otherRestrictions
Access Constraints: none. Use Constraints: Please follow guidelines at: http://www.bco-dmo.org/terms-use Distribution liability: Under no circumstances shall BCO-DMO be liable for any direct, incidental, special, consequential, indirect, or punitive damages that result from the use of, or the inability to use, the materials in this data submission. If you are dissatisfied with any materials in this data submission your sole and exclusive remedy is to discontinue use.
The Use of Nitrogen Isotopes of Amino Acids To Understand Marine Sedimentary 15N Records
https://osprey.bco-dmo.org/project/704684
The Use of Nitrogen Isotopes of Amino Acids To Understand Marine Sedimentary 15N Records
<p>The bioavailability of nutrients plays a crucial role in oceanic biological productivity, the carbon cycle, and climate change. The global ocean inventory of nitrogen (N) is determined by the balance of N-fixation (sources) and denitrification (sinks). In this three-year project, a researcher from the University of California, Santa Cruz, will focus on developing compound-specific N isotope (d15N) analysis of amino acids as a new tool for understanding N source and transformation of organic matter in paleo-reservoirs. The offsets in the isotopic ratios of individual amino acid groups may yield information about trophic transfer, heterotrophic microbial reworking, and autotrophic versus heterotrophic sources. By measuring and comparing the bulk and amino acid d15N in size-fractioned samples from plankton tows, sediments traps, and multi-cores in oxic and suboxic depositional environments, the researcher will: (1) Provide a proxy of the d15N of average exported photoautotrophic organic matter; and (2) Provide a new level of detail into sedimentary organic N degradation and preservation.</p>
<p>Broader impacts:<br />
This project will improve understanding of the fundamental underpinnings and behaviors of d15N amino acid patterns and how they behave in contrasting sedimentary environments, while also developing a potential paleoceanographic proxy. Funding will support a graduate student and undergraduate research at the institution. The researcher will also conduct community outreach in the form of a workshop/tutorial on the proxy development.</p>
Amino Acid Sediment 15N
largerWorkCitation
project
eng; USA
oceans
California Coast Littoral Waters
-125.0088
-116.4835
31.9339
42.9567
2009-01-01
2013-12-31
California Margin , Santa Barbara Basin , CA current system, Eastern Tropical Pacific
0
BCO-DMO catalogue of parameters from Amino acid isotopes from mussels along the California margin from 2009-2013 (Amino Acid Sediment 15N project)
Biological and Chemical Oceanography Data Management Office (BCO-DMO)
Unavailable
508-289-2009
WHOI MS#36
Woods Hole
MA
02543
USA
info@bco-dmo.org
http://www.bco-dmo.org
Monday - Friday 8:00am - 5:00pm
For questions regarding this resource, please contact BCO-DMO via the email address provided.
pointOfContact
http://lod.bco-dmo.org/id/dataset-parameter/714051.rdf
Name: Site
Units: unitless
Description: <p>Site where sampling occurred</p>
http://lod.bco-dmo.org/id/dataset-parameter/714052.rdf
Name: Identifier
Units: unitless
Description: <p>Sample ID</p>
http://lod.bco-dmo.org/id/dataset-parameter/714053.rdf
Name: Habitat_Type
Units: unitless
Description: <p>Description of habitat</p>
http://lod.bco-dmo.org/id/dataset-parameter/714054.rdf
Name: n
Units: count
Description: <p>Number of samples analyzed</p>
http://lod.bco-dmo.org/id/dataset-parameter/714055.rdf
Name: Latitude_degMin
Units: Degree minutes
Description: <p>Latitude in degree minutes</p>
http://lod.bco-dmo.org/id/dataset-parameter/714056.rdf
Name: Longitude_degMin
Units: Degree minutes
Description: <p>Longitude in degree minutes</p>
http://lod.bco-dmo.org/id/dataset-parameter/714057.rdf
Name: lat
Units: Decimal degrees
Description: <p>Latitude in decimal degrees</p>
http://lod.bco-dmo.org/id/dataset-parameter/714058.rdf
Name: lon
Units: Decimal degrees
Description: <p>Longitude in decimal degress</p>
http://lod.bco-dmo.org/id/dataset-parameter/714059.rdf
Name: Bulk_d15N
Units: per mil
Description: <p>15N/14N isotopic ratio</p>
http://lod.bco-dmo.org/id/dataset-parameter/714060.rdf
Name: Bulk_d15N_stdev
Units: per mil
Description: <p>Standard deviation of isotopic ratio</p>
http://lod.bco-dmo.org/id/dataset-parameter/714061.rdf
Name: Bulk_d13C
Units: per mil
Description: <p>13C/12C isotopic ratio</p>
http://lod.bco-dmo.org/id/dataset-parameter/714062.rdf
Name: Bulk_d13C_stdev
Units: per mil
Description: <p>Standard deviation of isotopic ratio</p>
http://lod.bco-dmo.org/id/dataset-parameter/714063.rdf
Name: Average_Trophic
Units: per mil
Description: <p>Average d15N values of [ Alanine, Aspartic Acid, Glutamic Acid, Leucine, Isoleucine, Proline, Valine ]</p>
http://lod.bco-dmo.org/id/dataset-parameter/714064.rdf
Name: Average_Source
Units: per mil
Description: <p>Average d15N values of [ Glycine, Lysine, Phenylalanine, Serine, Tyrosine ]</p>
http://lod.bco-dmo.org/id/dataset-parameter/714065.rdf
Name: Trophic_Position
Units: unitless
Description: <p>Trophic Position; { [ (d15N Glutamic Acid - d15N Phenylalanine) - 3.4 ] / 7.6 } + 1</p>
http://lod.bco-dmo.org/id/dataset-parameter/714066.rdf
Name: Ala
Units: per mil
Description: <p>Alanine 15N or 13C value</p>
http://lod.bco-dmo.org/id/dataset-parameter/714067.rdf
Name: Ala_stdev
Units: per mil
Description: <p>Alanine standard deviation</p>
http://lod.bco-dmo.org/id/dataset-parameter/714068.rdf
Name: Asx
Units: per mil
Description: <p>Asn+Asp 15N or 13C peak values</p>
http://lod.bco-dmo.org/id/dataset-parameter/714069.rdf
Name: Asx_stdev
Units: per mil
Description: <p>Asx standard deviation</p>
http://lod.bco-dmo.org/id/dataset-parameter/714070.rdf
Name: Glx
Units: per mil
Description: <p>Gln+Glu 15N or 13C peak values</p>
http://lod.bco-dmo.org/id/dataset-parameter/714071.rdf
Name: Glx_stdev
Units: per mil
Description: <p>Glx standard deviation</p>
http://lod.bco-dmo.org/id/dataset-parameter/714072.rdf
Name: Gly
Units: per mil
Description: <p>Glycine 15N or 13C value</p>
http://lod.bco-dmo.org/id/dataset-parameter/714073.rdf
Name: Gly_stdev
Units: per mil
Description: <p>Glycine standard deviation</p>
http://lod.bco-dmo.org/id/dataset-parameter/714074.rdf
Name: Ile
Units: per mil
Description: <p>Isoleucine 15N or 13 C value</p>
http://lod.bco-dmo.org/id/dataset-parameter/714075.rdf
Name: Ile_stdev
Units: per mil
Description: <p>Isoleucine standard deviation</p>
http://lod.bco-dmo.org/id/dataset-parameter/714076.rdf
Name: Leu
Units: per mil
Description: <p>Leucine 15N or 13C value</p>
http://lod.bco-dmo.org/id/dataset-parameter/714077.rdf
Name: Leu_stdev
Units: per mil
Description: <p>Leucine standard deviation</p>
http://lod.bco-dmo.org/id/dataset-parameter/714078.rdf
Name: Lys
Units: per mil
Description: <p>Lysine 15N or 13C value</p>
http://lod.bco-dmo.org/id/dataset-parameter/714079.rdf
Name: Lys_stdev
Units: per mil
Description: <p>Lysine standard deviation</p>
http://lod.bco-dmo.org/id/dataset-parameter/714080.rdf
Name: Phe
Units: per mil
Description: <p>Phenylalanine 15N or 13C value</p>
http://lod.bco-dmo.org/id/dataset-parameter/714081.rdf
Name: Phe_stdev
Units: per mil
Description: <p>Phenylalanine standard deviation</p>
http://lod.bco-dmo.org/id/dataset-parameter/714082.rdf
Name: Pro
Units: per mil
Description: <p>Proline 15N or 13C value</p>
http://lod.bco-dmo.org/id/dataset-parameter/714083.rdf
Name: Pro_stdev
Units: per mil
Description: <p>Proline standard deviation</p>
http://lod.bco-dmo.org/id/dataset-parameter/714084.rdf
Name: Ser
Units: per mil
Description: <p>Serine 15N or 13C value</p>
http://lod.bco-dmo.org/id/dataset-parameter/714085.rdf
Name: Ser_stdev
Units: per mil
Description: <p>Serine standard deviation</p>
http://lod.bco-dmo.org/id/dataset-parameter/714086.rdf
Name: Thr
Units: per mil
Description: <p>Threonine 15N or 13C value</p>
http://lod.bco-dmo.org/id/dataset-parameter/714087.rdf
Name: Thr_stdev
Units: per mil
Description: <p>Threonine standard deviation</p>
http://lod.bco-dmo.org/id/dataset-parameter/714088.rdf
Name: Tyr
Units: per mil
Description: <p>Tyrosine 15N or 13C value</p>
http://lod.bco-dmo.org/id/dataset-parameter/714089.rdf
Name: Tyr_stdev
Units: per mil
Description: <p>Tyrosine standard deviation</p>
http://lod.bco-dmo.org/id/dataset-parameter/714090.rdf
Name: Val
Units: per mil
Description: <p>Valine 15N or 13C value</p>
http://lod.bco-dmo.org/id/dataset-parameter/714091.rdf
Name: Val_stdev
Units: per mil
Description: <p>Valine standard deviation</p>
http://lod.bco-dmo.org/id/dataset-parameter/714092.rdf
Name: element
Units: unitless
Description: <p>Isotopic element; Nitrogen AA or Carbon AA</p>
GB/NERC/BODC > British Oceanographic Data Centre, Natural Environment Research Council, United Kingdom
Biological and Chemical Oceanography Data Management Office (BCO-DMO)
Unavailable
508-289-2009
WHOI MS#36
Woods Hole
MA
02543
USA
info@bco-dmo.org
http://www.bco-dmo.org
Monday - Friday 8:00am - 5:00pm
For questions regarding this resource, please contact BCO-DMO via the email address provided.
pointOfContact
9914
https://darchive.mblwhoilibrary.org/bitstream/1912/24249/1/dataset-713831_amino-acid-isotopes-mussels__v1.tsv
download
https://doi.org/10.1575/1912/bco-dmo.713831.1
download
onLine
dataset
<p><strong>Sampling</strong></p>
<p>Mussels were collected in the winter (Dec – Feb) of 2009–2010. Sites were chosen to be approximately evenly distributed along the CA coastline, with ,80 km geographic separation between each</p>
<p>sampling site. Our main goal here was to sample mussels from a wide geographic range across the CCS, although for observing finer scale local or regional variations, a finer-scale sampling strategy would like be required. Typically 5 individual mussels were collected from each site, all between 30–40 mm maximum shell length, which were immediately placed on dry ice until further preparation. The adductor muscle of each individual was dissected for analysis. This tissue was selected because isotopic values in muscle tissue have shown relatively long turnover times; based on past growth data, mussels of this size would be expected to integrate approximately annual variability in suspended food source isotopic values for each location sampled.</p>
<p>The dissected adductor tissue was carefully separated from other tissue types, rinsed with deionized water, refrozen, and then freeze-dried for 48 hrs. Lipids were removed &nbsp;using petroleum ether in a Dionex Accelerated Solvent Extractor (Bannockburn, IL). Finally, in preparation for CSI-AA, composite samples were made from a subset of 13 collection sites. For each location chosen for CSI-AA (based on the bulk d15N record), 160.05 mg of lyophilized tissue was weighed and combined for each individual mussel (n = 5).</p>
<p><strong>Elemental and Bulk Isotopic Analyses</strong></p>
<p>Stable carbon (d13C) and nitrogen (d15N) isotope ratios were determined via elemental analyzer isotope ratio mass spectrometry (EA-IRMS) at the University of California, Santa Cruz, Stable Isotope Laboratory (UCSC-SIL; http://emerald.ucsc.edu/~silab/). Approximately 1 mg of each dry isolated DOM sample was weighed into tin capsules (Costec, 5 x 9 mm) for analysis. EA-IRMS analysis was conducted using a Carlo Erba CHNS-O EA1108-elemental analyzer interfaced via a ConFlo III device with a ThermoFinnigan Delta Plus XP isotope ratio mass spectrometer (Thermo Fisher Scientific). &nbsp;Standards, EA-IRMS protocols, and correction routines followed standard UCSC-SIL protocols. Analytical uncertainties of n=3 replicate measurements of isotopic standards ranged from ± 0.05 to 0.1‰ for both d13C and d15N. Carbon to nitrogen elemental ratios were similarly determined by elemental analysis. The presented ratios are atomic ratios (C/N)a normalized to the mass of C and N, but have been abbreviated as C/N throughout.</p>
<p><strong>Compound-specific amino acid Isotopic 13C Analyses</strong></p>
<p>Individual AA d13C values were measured as trifluoroacetyl isopropyl ester (TFA-IP) AA derivatives, after acid hydrolysis. Samples were hydrolyzed by adding 2.5 mg homogenized composite muscle tissue to 1 ml of 6 N HCl, and heating for 20 h at 110 deg C under nitrogen. After drying, AA isopropyl esters were prepared with a 1:5 mixture of AcCl:2-propanol (110 deg C, 60 min) and then acylated using a 1:3 mixture of di chloro - methane (DCM) and trifluoroacetic anhydride (TFAA) (100 deg C, 15 min). Derivatized AAs were dissolved in DCM to a final ratio of approximately 2.5 mg of original tissue to 250 ul DCM for injection on the gas chromatograph IRMS (GC-IRMS) system.</p>
<p>Isotopic analysis was conducted on a Thermo Trace GC Ultra with inline oxidation and reduction furnaces, coupled to a ThermoFinnigan Delta Plus XP IRMS, equipped with a CTC Analytics autosampler.Derivatives (1 ul) were injected (250 deg C constant temperature) onto an Agilent DB-5 column (50 m × 0.32 mm ID × 0.52 um film thickness), with a He carrier flow rate of 2 ml min−1 (constant flow). Separations were achieved with a 4-ramp oven program: 52 deg C, 2 min hold; ramp 1 = 15 deg C min−1 to 75 deg C, hold for 2 min; ramp 2 = 4 deg C min−1 to 185 deg C, hold for 2 min; ramp 3 = 4 deg C min−1 to 200 deg C; ramp 4 = 30 deg C min−1 to 240 deg C, hold for 5 min.</p>
<p>This method allowed for d13C determination of the following AAs in mussel tissue: non-essential AAs alanine (Ala), aspartic acid + asparagine (Asp), glutamic acid + glutamine (Glu), glycine (Gly), proline (Pro), serine (Ser), and tyrosine Tyr); and essential AAs leucine (Leu), isoleucine (Ile), valine (Val), phenylalanine (Phe), and threonine (Thr). Acid hydrolysis destroys tryptophan and cystine, so these were not detected, and it also deaminates\ asparagine to aspartic acid, and glutamine to glutamic acid. While the abbreviations Glx and Asx\ are sometimes used to denote the combined Gln+Glu and Asn+Asp peaks, in order to correspond better with extant CSI-AA literature, we have elected to simply use Asp and Glu abbreviations, as defined above.</p>
<p>All samples were analyzed on the GC-IRMS in triplicate, and measured AA d13C values were corrected for the C added during derivatization, following the approach of Silfer et al. (1991). Reproducibility for tissue samples was typically less than &lt;0.3‰ (n = 3). The average mean deviation for all tissue sample replicates was 0.4‰.</p>
<p><strong>Compound-specific amino acid Isotopic 15N Analyses</strong></p>
<p>Amino acid d15N values were measured as Trifluoroacetyl isopropyl ester (TFA-IP) AA derivatives, following protocols described in detail elsewhere (e.g.,. Briefly, samples were hydrolyzed (6 N HCl, 20 hr at 110uC) under nitrogen, and TFA derivatives subsequently prepared from free AA using a modified version of the protocol described by Silfer (Silfer et al. 1991): isopropyl esters were made with a 1:5 mixture of Acetyl Chloride (AcCl):2-propanol (110uC, 60 minutes), and then acylated using a 1:3 mixture of Dichloromethane:Trifluroacetyl acetate (DCM:TFAA) (100uC, 15 minutes). Derivatized AAs were dissolved in DCM to a final ratio of approximately 4 mg of original tissue to 250 ml DCM. After derivatization, samples were analyzed by a Varian gaschromatograph coupled to a Finnegan Delta-Plus isotope ratiomass spectrometer (GC-IRMS). AAs were separated using a 50 m, 0.32 ID Hewlett Packard Ultra-1 column with 1 mm film thickness. Under our analytical conditions, d15N values could be reproducibly measured for alanine (Ala), aspartic acid + asparagine (Asp), glutamic acid + glutamine (Glu), leucine (Leu), isoleucine (Ile), proline (Pro), valine (Val), glycine (Gly), lysine (Lys), serine (Ser), phenylalanine (Phe), threonine (Thr), and tyrosine (Tyr) (Fig. S4). Most AAs were measured with a standard error of ,1.0% (based on n = 4 injections), and the average mean deviations for individual AA d15N measurements across all tissue sample replicates was 0.5%.</p>
Specified by the Principal Investigator(s)
<p>Thermo-Finnigan Isodat software and Microsoft Excel 2013.</p>
<p><strong>BCO-DMO Processing:</strong><br />
- modified parameter names to conform with BCO-DMO naming conventions;<br />
- replaced missing data/blanks&nbsp;with "nd" ("no data")</p>
Specified by the Principal Investigator(s)
asNeeded
7.x-1.1
Biological and Chemical Oceanography Data Management Office (BCO-DMO)
Unavailable
508-289-2009
WHOI MS#36
Woods Hole
MA
02543
USA
info@bco-dmo.org
http://www.bco-dmo.org
Monday - Friday 8:00am - 5:00pm
For questions regarding this resource, please contact BCO-DMO via the email address provided.
pointOfContact
Carlo Erba CHNS-O EA1108-elemental analyzer
Carlo Erba CHNS-O EA1108-elemental analyzer
PI Supplied Instrument Name: Carlo Erba CHNS-O EA1108-elemental analyzer PI Supplied Instrument Description:Interfaced via a ConFlo III device with a ThermoFinnigan Delta Plus XP isotope ratio mass spectrometer (Thermo Fisher Scientific) Instrument Name: Elemental Analyzer Instrument Short Name: Instrument Description: Instruments that quantify carbon, nitrogen and sometimes other elements by combusting the sample at very high temperature and assaying the resulting gaseous oxides. Usually used for samples including organic material. Community Standard Description: http://vocab.nerc.ac.uk/collection/L05/current/LAB01/
Trace GC, interfaced via a ConFlo III and GCC device
Trace GC, interfaced via a ConFlo III and GCC device
PI Supplied Instrument Name: Trace GC, interfaced via a ConFlo III and GCC device PI Supplied Instrument Description:Used with with a ThermoFinnigan Delta Plus XP isotope ratio mass spectrometer (Thermo Fisher Scientific) Instrument Name: Gas Chromatograph Instrument Short Name:Gas Chromatograph Instrument Description: Instrument separating gases, volatile substances, or substances dissolved in a volatile solvent by transporting an inert gas through a column packed with a sorbent to a detector for assay. (from SeaDataNet, BODC) Community Standard Description: http://vocab.nerc.ac.uk/collection/L05/current/LAB02/
hermoFinnigan Delta Plus XP isotope ratio mass spectrometer (Thermo Fisher Scientific)
hermoFinnigan Delta Plus XP isotope ratio mass spectrometer (Thermo Fisher Scientific)
PI Supplied Instrument Name: hermoFinnigan Delta Plus XP isotope ratio mass spectrometer (Thermo Fisher Scientific) PI Supplied Instrument Description:Used with Trace GC, interfaced via a ConFlo III and GCC device Instrument Name: Isotope-ratio Mass Spectrometer Instrument Short Name:IR Mass Spec; IRMS Instrument Description: The Isotope-ratio Mass Spectrometer is a particular type of mass spectrometer used to measure the relative abundance of isotopes in a given sample (e.g. VG Prism II Isotope Ratio Mass-Spectrometer). Community Standard Description: http://vocab.nerc.ac.uk/collection/L05/current/LAB16/
Deployment: McCarthy_2009
McCarthy_2009
shoreside Calif_shore
shoreside
McCarthy_2009
Matthew D. McCarthy
University of California-Santa Cruz
shoreside Calif_shore
shoreside