Dataset title: Dissolved Organic Carbon Stable Isotopes and Lignin Phenols from Everglades National Park (FCE LTER), South Florida, USA, January 2018 - ongoing Dataset ID: doi:10.6073/pasta/b46c5271c52e96907fb22514aa2843ce Dataset Creator Name: Dr. Chris Osburn Position: Lead PI Researcher Organization: North Carolina State University Email: closburn@ncsu.edu Metadata Provider Organization: Florida Coastal Everglades LTER Address: Florida International University 11200 SW 8th Street, OE 148 Miami, FL 33199 USA Phone: 305-348-6054 Email: fcelter@fiu.edu URL: https://fcelter.fiu.edu Dataset Abstract Estuarine and coastal waters are home to more than half of the world's population and provide many social and economic benefits to society. Additionally, these ecosystems are sites of intense carbon cycling and are under pressure from changing land uses and climate. DOM is functionally defined as the OM passed through a 0.7 µm borosilicate glass fiber filter (GF/F) and is primarily made up of viruses, bacteria, colloids, lignins, humic substances, organic acids, and other small organic compounds (Repeta, 2015). Approximately 50% of DOM is generally considered to be dissolved organic carbon (DOC) (Stedmon and Nelson, 2015). Thus, DOC can be used as a proxy for quantifying DOM. Whole water samples were collected monthly from three different transects representing marl-dominated (Taylor Slough), peat-dominated (Shark River Slough), and seagrass-dominated (Florida Bay) environments. The water was filtered using 0.7 µm porosity glass fiber filters, then acidified to pH 2 and stored at 4°C until processing occurred. DOC concentration and the stable isotope ratio of DOC (δ13C-DOC) was measured on an OI Analytical TOC analyzer coupled to an Agilent Delta V plus Isotope Ratio Mass Spectrometer using high-temperature combustion (Lalonde et al., 2014). Dissolved vanillyl (vanillin, acetovanillone, vanillin acid), syringyl (syringaldehyde, acetosyringone, syringic acid), cinnamyl (p-coumaric acid, ferulic acid), and 3,5-hydroxybenzoic acid lignin phenols were isolated from ~2 liters of 0.7 µm filtered, acidified (pH 2) water by solid phase extraction (SPE) using 1g PPL cartridges that were pre-conditioned using pH 2 water and methanol. SPE cartridges were extracted using methanol, then dried and redissolved in 2N NaOH. Cupric oxide oxidation and liquid-liquid ethyl acetate extraction were used to extract lignin oxidation products (Hedges and Ertel, 1982, modified by Goñi and Hedges, 1995; Louchouarn et al., 2000; Goñi and Montgomery, 2000; Benner and Kaiser, 2011). Briefly, NaOH dissolved samples were added to teflon vessels with CuO, ferrous ammonium sulfate, and glucose then oxidized in a muffle furnace for 150 minutes at 155C (5C/min ramp rate). Samples were then rapidly cooled, vortexed then transferred to centrifuge tubes and centrifuged, then the supernatant was transferred to a new centrifuge tube and internal lignin standards are added and then supernatant is acidified. Ethyl acetate is added to preform liquid-liquid extraction. Ethyl acetate extract is dried over a sodium sulfate column, then extracts are dried. Extracts are redissolved in pyridine then derivatized using BSTFA in a muffle furnace for 1 hr at 60C. Dertivatized samples are then dried and redissolved in dichloromethane then immediately quantified using an Agilent Gas Chromatograph-Mass Spectrometer (GC-MS) according to the method provided by Kaiser and Benner (2012). References: Benner, R., & Kaiser, K. (2011). Biological and photochemical transformations of amino acids and lignin phenols in riverine dissolved organic matter. Biogeochemistry, 102(1) 209-222. Goñi, M. A., & Hedges, J. I. (1995). Sources and reactivities of marine-derived organic matter in coastal sediments as determined by alkaline CuO oxidation. Geochimica et Cosmochimica Acta, 59(14), 2965-2981. Goñi, M. A., & Montgomery, S. (2000). Alkaline CuO oxidation with a microwave digestion system: Lignin analyses of geochemical samples. Analytical chemistry, 72(14), 3116-3121. Hedges, J. I., & Ertel, J. R. (1982). Characterization of lignin by gas capillary chromatography of cupric oxide oxidation products. Analytical Chemistry, 54(2), 174-178. Kaiser, K., & Benner, R. (2012). Characterization of lignin by gas chromatography and mass spectrometry using a simplified CuO oxidation method. Analytical chemistry, 84(1), 459-464. Lalonde, K., Middlestead, P., & Gélinas, Y. (2014). Automation of 13C/12C ratio measurement for freshwater and seawater DOC using high temperature combustion. Limnology and Oceanography: Methods, 12(12), 816-829. Louchouarn, P.; Opsahl, S.; Benner, R. 2000. Isolation and quantification of dissolved lignin from natural waters using solid phase extraction and GC/MS. Analytical Chem. 2000, 2780-2787. Repeta, D. J. (2015). Chemical characterization and cycling of dissolved organic matter. In Biogeochemistry of marine dissolved organic matter (pp. 21-63). Academic Press. Stedmon, C. A., & Nelson, N. B. (2015). The optical properties of DOM in the ocean. In Biogeochemistry of marine dissolved organic matter (pp. 481-508). Academic Press. Geographic Coverage Bounding Coordinates Geographic description: SRS1d West bounding coordinate: -80.654 East bounding coordinate: -80.654 North bounding coordinate: 25.7463 South bounding coordinate: 25.7463 Geographic description: SRS2 West bounding coordinate: -80.78520692 East bounding coordinate: -80.78520692 North bounding coordinate: 25.54972811 South bounding coordinate: 25.54972811 Geographic description: SRS3 West bounding coordinate: -80.85327617 East bounding coordinate: -80.85327617 North bounding coordinate: 25.46820617 South bounding coordinate: 25.46820617 Geographic description: SRS4 West bounding coordinate: -80.96431016 East bounding coordinate: -80.96431016 North bounding coordinate: 25.40976421 South bounding coordinate: 25.40976421 Geographic description: SRS5 West bounding coordinate: -81.03234716 East bounding coordinate: -81.03234716 North bounding coordinate: 25.37702258 South bounding coordinate: 25.37702258 Geographic description: SRS6 West bounding coordinate: -81.07794623 East bounding coordinate: -81.07794623 North bounding coordinate: 25.36462994 South bounding coordinate: 25.36462994 Geographic description: TS/Ph1a West bounding coordinate: -80.59029790000001 East bounding coordinate: -80.59029790000001 North bounding coordinate: 25.42388762 South bounding coordinate: 25.42388762 Geographic description: TS/Ph2 West bounding coordinate: -80.60690341 East bounding coordinate: -80.60690341 North bounding coordinate: 25.40357188 South bounding coordinate: 25.40357188 Geographic description: TS/Ph3 West bounding coordinate: -80.66271768 East bounding coordinate: -80.66271768 North bounding coordinate: 25.25240534 South bounding coordinate: 25.25240534 Geographic description: TS/Ph6a West bounding coordinate: -80.6490792 East bounding coordinate: -80.6490792 North bounding coordinate: 25.21418102 South bounding coordinate: 25.21418102 Geographic description: TS/Ph7a West bounding coordinate: -80.63910514 East bounding coordinate: -80.63910514 North bounding coordinate: 25.19080491 South bounding coordinate: 25.19080491 Geographic description: TS/Ph9 West bounding coordinate: -80.48978207 East bounding coordinate: -80.48978207 North bounding coordinate: 25.17692874 South bounding coordinate: 25.17692874 Geographic description: TS/Ph10 West bounding coordinate: -80.68097374 East bounding coordinate: -80.68097374 North bounding coordinate: 25.02476744 South bounding coordinate: 25.02476744 Geographic description: TS/Ph11 West bounding coordinate: -80.93798347 East bounding coordinate: -80.93798347 North bounding coordinate: 24.91293492 South bounding coordinate: 24.91293492 Temporal Coverage Start Date: 2018-01-08 End Date: 2022-10-31 Data Table Entity Name: FCE1240_DON Entity Description: FCE LTER dissolved carbon and lignin and stable carbon isotopes Object Name: FCE1240_DON.csv Data Format Number of Header Lines: 1 Attribute Orientation: column Field Delimiter: , Number of Records: Attributes Attribute Name: SITENAME Attribute Label: Site Attribute Definition: Name of LTER Sampling Site Location Storage Type: string Measurement Scale: SRS1d= Shark River Slough site 1d SRS2= Shark River Slough site 2 SRS3= Shark River Slough site 3 SRS4= Shark River Slough site 4 SRS5= Shark River Slough site 5 SRS6= Shark River Slough site 6 TS/Ph1a= Taylor Slough site 1a TS/Ph2= Taylor Slough site 2 TS/Ph3= Taylor Slough site 3 TS/Ph6a= Taylor Slough site 6a TS/Ph7a= Taylor Slough site 7a TS/Ph9= Taylor Slough site 9 TS/Ph10= Taylor Slough site 10 TS/Ph11= Taylor Slough site 11 Missing Value Code: -9999 (Sample not currently analyzed) Attribute Name: Month Attribute Label: Month Attribute Definition: Month of the year Storage Type: float Measurement Scale: Units: month Number Type: integer Missing Value Code: Attribute Name: Year Attribute Label: Year Attribute Definition: Sampling year Storage Type: dateTime Measurement Scale: Missing Value Code: Attribute Name: Date Attribute Label: Date Attribute Definition: Date of Sampling Storage Type: dateTime Measurement Scale: Missing Value Code: Attribute Name: Sample_Name Attribute Label: Sample_Name Attribute Definition: Lab Sample identification name Storage Type: string Measurement Scale: text Missing Value Code: Attribute Name: DOC_mg_L Attribute Label: DOC_mg_L Attribute Definition: Dissolved organic carbon concentration in mg per liter Storage Type: float Measurement Scale: Units: milligramPerLiter Number Type: real Missing Value Code: -9999 (Sample not currently analyzed) Attribute Name: d13C_DOC_per_mille Attribute Label: d13C_DOC_per_mille Attribute Definition: Stable isotope ratio of dissolved organic carbon Storage Type: float Measurement Scale: Units: permil Number Type: real Missing Value Code: -9999 (Sample not currently analyzed) Attribute Name: Vanillin_ug_L Attribute Label: Vanillin_ug_L Attribute Definition: Concentration of vanillin lignin phenol Storage Type: float Measurement Scale: Units: microgramPerLiter Number Type: real Missing Value Code: -9999 (Sample not currently analyzed) Attribute Name: Acetovanillone_ug_L Attribute Label: Acetovanillone_ug_L Attribute Definition: Concentration of acetovanillone lignin phenol Storage Type: float Measurement Scale: Units: microgramPerLiter Number Type: real Missing Value Code: -9999 (Sample not currently analyzed) Attribute Name: Vanillic_acid_ug_L Attribute Label: Vanillic_acid_ug_L Attribute Definition: Concentration of vanillic acid lignin phenol Storage Type: float Measurement Scale: Units: microgramPerLiter Number Type: real Missing Value Code: -9999 (Sample not currently analyzed) Attribute Name: Syringaldehyde_ug_L Attribute Label: Syringaldehyde_ug_L Attribute Definition: Concentration of syringaldehyde lignin phenol Storage Type: float Measurement Scale: Units: microgramPerLiter Number Type: real Missing Value Code: -9999 (Sample not currently analyzed) Attribute Name: Acetosyringone_ug_L Attribute Label: Acetosyringone_ug_L Attribute Definition: Concentration of acetosyringone lignin phenol Storage Type: float Measurement Scale: Units: microgramPerLiter Number Type: real Missing Value Code: -9999 (Sample not currently analyzed) Attribute Name: Syringic_acid_ug_L Attribute Label: Syringic_acid_ug_L Attribute Definition: Concentration of syringic acid lignin phenol Storage Type: float Measurement Scale: Units: microgramPerLiter Number Type: real Missing Value Code: -9999 (Sample not currently analyzed) Attribute Name: P_coumaric_acid_ug_L Attribute Label: P_coumaric_acid_ug_L Attribute Definition: Concentration of p-coumaric acid lignin phenol Storage Type: float Measurement Scale: Units: microgramPerLiter Number Type: real Missing Value Code: -9999 (Sample not currently analyzed) Attribute Name: Ferulic_acid_ug_L Attribute Label: Ferulic_acid_ug_L Attribute Definition: Concentration of ferulic acid lignin phenol Storage Type: float Measurement Scale: Units: microgramPerLiter Number Type: real Missing Value Code: -9999 (Sample not currently analyzed) Attribute Name: 3_5_dihydroxy_benzoic_acid Attribute Label: 3_5_dihydroxy_benzoic_acid Attribute Definition: Concentration of 3,5-dihydroxy-benzoic acid lignin phenol Storage Type: float Measurement Scale: Units: microgramPerLiter Number Type: real Missing Value Code: -9999 (Sample not currently analyzed) Methods Method Step Description Method Step Description Method Step Description Dissolve lignin-derived phenols were isolated from FCE waters using solid-phase extraction (SPE) with 6 mL Agilent Bond Elut PPL filters, which contain 1 g of styrene-divinylbenzene polymer with a non-polar surface (Louchouarn et al., 2000; Arellano et al., 2018). Briefly, the filtered and acidified sample was conditioned with methanol and acidified Milli-Q Ultrapure water to ensure that the sorbent was saturated before the SPE (Louchouarn et al., 2000). An approximately 2 L acidifed sample was pumped through a PPL filter using a peristaltic pump at a rate of 10 mL min-1. The PPL cartridge was rinsed with 18 mL of acidified (pH 2) Milli-Q water to remove any remaining salts (Louchouarn et al., 2000). The PPL cartridge was then dried under ultra-high purity nitrogen gas (10-12 psi) and then eluted into borosilicate glass vials that were baked at 450C for 5 hours, using 9 mL HPLC grade methanol (Arellano et al., 2018). The cupric oxide oxidation (CuO) and liquid-liquid extraction methods (Hedges and Ertel, 1982, modified by Goñi and Hedges, 1995; Louchouarn et al., 2000; Goñi and Montgomery, 2000; Benner and Kaiser, 2011, respectively) were used to extract lignin oxidation products. 500 mg of CuO, 100 mg of ferrous ammonium sulfate (FAS), and 10 mg of glucose was then added to 6 mL Salvillex Teflon reaction vessels. The methanol eluent was split into two subsamples and then dried using nitrogen. One subsample of dried eluent was then redissolved in 3.5 mL of nitrogen-sparged 2 N NaOH and added to the Teflon reaction vessel. The reaction vessel was then briefly sparged with nitrogen to remove excess oxygen and then capped. Samples were then oxidized in a muffle furnace for 150 minutes at 155C (5C min -1 ramp rate). Once oxidation was complete the Teflon vessels were cooled in an ice bath, then vortexed, transferred to polycarbonate tubes and centrifuged at 3200 rpm for 20 minutes. Supernatant was transferred to a new centrifuge tube then ethyl vanillin and t-cinnamic acid are added prior to liquid-liquid extraction to be used as recovery standards. Next the sample was acidified with 85% H3PO4 to pH 2 and then vortexed and centrifuged. Liquid-liquid extraction was completed by addition of 4 mL of ethyl acetate to the acidified sample, followed by vortex and centrifuge, then passed over a Na2SO4 column to remove residual water. Lignin liquid-liquid extracts were then dried under nitrogen, redissolved in 500 uL of pyridine, and stored at -20C until analysis. 50 uL of liquid-liquid extract is then derivatized using 50 uL of N,O-bis (trimethylsilyl) trifluoroacetamide (BSTFA) at 60°C for 1 hour. Derivatized samples were then dried under nitrogen, redissolved in 100 uL of dichloromethane and then measured and quantified immediately on an Agilent Gas Chromatograph-Mass Spectrometer (GC-MS) according to the method provided by Kaiser and Benner (2012). Relative response factors were calculated using t-cinnamic acid to account for instrument variability. Concentrations of dissolved lignin oxidation products are calculated using calibration curves of 9 phenols (vanillyl, syringyl, cinnamyl, and 3,5 Bd) over the range of expected values. Distribution Online distribution: https://pasta.lternet.edu/package/data/eml/knb-lter-fce/1240/2/95397937db4c76fa457813869262322b Intellectual Rights This information is released under the Creative Commons license - Attribution - CC BY (https://creativecommons.org/licenses/by/4.0/). The consumer of these data ("Data User" herein) is required to cite it appropriately in any publication that results from its use. The Data User should realize that these data may be actively used by others for ongoing research and that coordination may be necessary to prevent duplicate publication. The Data User is urged to contact the authors of these data if any questions about methodology or results occur. Where appropriate, the Data User is encouraged to consider collaboration or co-authorship with the authors. The Data User should realize that misinterpretation of data may occur if used out of context of the original study. While substantial efforts are made to ensure the accuracy of data and associated documentation, complete accuracy of data sets cannot be guaranteed. All data are made available "as is." The Data User should be aware, however, that data are updated periodically and it is the responsibility of the Data User to check for new versions of the data. The data authors and the repository where these data were obtained shall not be liable for damages resulting from any use or misinterpretation of the data. Thank you. Dataset Keywords dissolved organic carbon carbon carbon cycling stable isotopes concentration marshes mangroves rivers estuaries dissolved lignin phenols LTER FCE LTER Florida Coastal Everglades LTER long-term monitoring Everglades National Park organic matter Maintenance knb-lter-fce.1240.2: Added new data, updated metada Dataset Contact Name: Dr. Chris Osburn Position: Lead PI Researcher Organization: North Carolina State University Email: closburn@ncsu.edu