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Contribution of ketone/aldehyde-containing compounds to the composition and optical properties of Suwannee River fulvic acid revealed by ultrahigh resolution mass spectrometry and deuterium labeling


A prior method of mass labeling ketone-/aldehyde-containing species in natural dissolved organic matter (DOM) is further developed and applied. This application involved the treatment of Suwannee River fulvic acid (SRFA) with increasing concentrations of sodium borodeuteride (NaBD4), followed by detection of reduced species via negative mode electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI FTICR MS). The extent of reduction, as determined by ESI FTICR MS, resulting from increasing concentrations of NaBD4 correlated well with changes in the absorption and emission spectra of the corresponding untreated and borodeuteride-reduced samples, providing evidence that ketone/aldehyde functional groups contribute substantially to the bulk optical properties of SRFA. Furthermore, the differences in the reactivity and abundance of ketone-/aldehyde-containing species for various regions in Van Krevelen plots were revealed, thus showing how this mass labeling method can be used to provide more detailed structural information about components within complex DOM samples than that provided by the determination and analysis of molecular formulae alone.

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  1. 1.

    Salonen K, Hammar T. On the importance of dissolved organic matter in the nutrition of zooplankton in some lake waters. Oecologia. 1986;68:246–53.

  2. 2.

    Blough NV, Del Vecchio R. Chromophoric DOM in the coastal environment. In: Hansell DA, Carlson CA, editors. Biogeochem. San Diego: Academic; 2002.

  3. 3.

    Coble PG. Marine optical biogeochemistry: the chemistry of ocean color. Chem Rev. 2007;107:402–18. https://doi.org/10.1021/cr050350+.

  4. 4.

    Zepp RG, Erickson DJ III, Paul ND, Sulzberger B. Interactive effects of solar UV radiation and climate change on biogeochemical cycling. Photochem Photobiol Sci. 2007;6:286. https://doi.org/10.1039/b700021a.

  5. 5.

    Morris DP, Hargreaves BR. The role of photochemical degradation of dissolved organic carbon in regulating the UV transparency of three lakes on the Pocono Plateau. Limnol Oceanogr. 1997;42:239–249.

  6. 6.

    Nelson NB, Siegel DA. Chapter 11 - Chromophoric DOM in the Open Ocean. In: Biogeochemistry of marine dissolved organic matter. 2002;pp 547–VI.

  7. 7.

    Zepp RG, Erickson DJ III, Paul ND, Sulzberger B. Interactive effects of solar UV radiation and climate change on biogeochemical cycling. Photochem Photobiol Sci. 2007:286–300. https://doi.org/10.1039/b700021a.

  8. 8.

    Scully NM, Cooper WJ, Tranvik LJ. Photochemical effects on microbial activity in natural waters: the interaction of reactive oxygen species and dissolved organic matter. FEMS Microbiol Ecol. 2003;46:353–7. https://doi.org/10.1016/S0168-6496(03)00198-3.

  9. 9.

    Berg SM, Whiting QT, Herrli JA, Winkels R, Wammer KH, Remucal CK. The role of dissolved organic matter composition in determining photochemical reactivity at the molecular level. Environ Sci Technol. 2019;53:11725–34.

  10. 10.

    Boyle ES, Guerriero N, Thiallet A, Del Vecchio R, Blough NV. Optical properties of humic substances and CDOM: relation to structure. Environ Sci Technol. 2009;43:2262–8.

  11. 11.

    Del Vecchio R, Blough NV. On the origin of the optical properties of humic substances. Environ Sci Technol. 2004;38:3885–91.

  12. 12.

    Schendorf TM, Del Vecchio R, Bianca M, Blough NV. Combined effects of pH and borohydride reduction on optical properties of humic substances (HS): a comparison of optical models. Environ Sci Technol. 2019;53:6310–9.

  13. 13.

    Sharpless CM, Blough NV. The importance of charge-transfer interactions in determining chromophoric dissolved organic matter (CDOM) optical and photochemical properties. Environ Sci Process Impacts. 2014;16:654–71.

  14. 14.

    Tossell JA. Quinone–hydroquinone complexes as model components of humic acids: theoretical studies of their structure, stability and visible–UV spectra. Geochim Cosmochim Acta. 2009;73:2023–33.

  15. 15.

    Balraj C, Satheshkumar A, Ganesh K, Elango KP. Charge transfer complexes of quinones in aqueous medium: spectroscopic and theoretical studies on interaction of cimetidine with novel substituted 1,4-benzoquinones and its application in colorimetric sensing of anions. Spectrochim Acta Part A Mol Biomol Spectrosc. 2013;114:256–66.

  16. 16.

    Abdulla HAN, Minor EC, Hatcher PG. Using two-dimensional correlations of 13C NMR and FTIR to investigate changes in the chemical composition of dissolved organic matter along an estuarine transect. Environ Sci Technol. 2010;44:8044–9.

  17. 17.

    Hertkorn N, Harir M, Koch BP, Michalke B, Schmitt-Kopplin P. High-field NMR spectroscopy and FTICR mass spectrometry: powerful discovery tools for the molecular level characterization of marine dissolved organic matter. Biogeosciences. 2013;10:1583–624.

  18. 18.

    Janot N, Reiller PE, Korshin GV. Using spectrophotometric titrations to characterize humic acid reactivity at environmental concentrations. Environ Sci Technol. 2010;44:6782–8.

  19. 19.

    Reemtsma T, These A, Springer A, Linscheid M. Differences in the molecular composition of fulvic acid size fractions detected by size-exclusion chromatography-on line Fourier transform ion cyclotron resonance (FTICR-) mass spectrometry. Water Res. 2008;42:63–72.

  20. 20.

    Leenheer JA, Wilson MA, Malcolm RL. Presence and potential significance of aromatic-ketone groups in aquatic humic substances. Org Geochem. 1987;11:273–80.

  21. 21.

    Macalady D, Walton-Day K. Redox chemistry and natural organic matter (NOM): geochemists’ dream, analytical chemists’ nightmare. In: Tratnyek PG, Grundi TJ, Haderlein SB, editors. Aquatic redox chemistry. ACS symposium series 1071. Washington, D. C: American Chemical Society; 2011. p. 85–111.

  22. 22.

    Ma J, Del Vecchio R, Golanoski KS, Boyle ES, Blough NV. Optical properties of humic substances and CDOM: effects of borohydride reduction. Environ Sci Technol. 2010;44:5395–402.

  23. 23.

    Zhang Y, Del Vecchio R, Blough NV. Investigating the mechanism of hydrogen peroxide photoproduction by humic substances. Environ Sci Technol. 2012;46:11836–43.

  24. 24.

    Andrew AA, Del Vecchio R, Subramaniam A, Blough NV. Chromophoric dissolved organic matter (CDOM) in the equatorial Atlantic Ocean: optical properties and their relation to CDOM structure and source. Mar Chem. 2013;148:33–43.

  25. 25.

    Aeschbacher M, Graf C, Schwarzenbach RP, Sander M. Antioxidant properties of humic substances. Environ Sci Technol. 2012;46:4916–25. https://doi.org/10.1021/es300039h.

  26. 26.

    Baluha DR, Blough NV, Del Vecchio R. Selective mass labeling for linking the optical properties of chromophoric dissolved organic matter to structure and composition via ultrahigh resolution electrospray ionization mass spectrometry. Environ Sci Technol. 2013;47:9891–7. https://doi.org/10.1021/es402400j.

  27. 27.

    Gonsior M, Schmitt-Kopplin P, Bastviken D. Depth-dependent molecular composition and photo-reactivity of dissolved organic matter in a boreal lake under winter and summer conditions. Biogeosciences. 2013;10:6945–56.

  28. 28.

    Gonsior M, Peake BM, Cooper WT, Podgorski D, D’Andrilli J, Dittmar T, et al. Characterization of dissolved organic matter across the Subtropical Convergence off the South Island, New Zealand. Mar Chem. 2011;123:99–110.

  29. 29.

    Hertkorn N, Frommberger M, Witt M, Koch BP, Schmitt-Kopplin P, Perdue EM. Natural organic matter and the event horizon of mass spectrometry. Anal Chem. 2008;80:8908–19.

  30. 30.

    Koch BP, Dittmar T, Witt M, Kattner G. Fundamentals of molecular formula assignment to ultrahigh resolution mass data of natural organic matter. Anal Chem. 2007;79:1758–63.

  31. 31.

    Stenson AC, Marshall AG, Cooper WT. Exact masses and chemical formulas of individual Suwannee River fulvic acids from ultrahigh resolution electrospray ionization Fourier transform ion cyclotron resonance mass spectra. Anal Chem. 2003;75:1275–84.

  32. 32.

    Cao D, Huang H, Hu M, Cui L, Geng F, Rao Z, et al. Comprehensive characterization of natural organic matter by MALDI- and ESI-Fourier transform ion cyclotron resonance mass spectrometry. Anal Chim Acta. 2015;866:48–58.

  33. 33.

    Mugo S, Bottaro C. Characterization of humic substances by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Rapid Commun Mass Spectrom. 2004;18:2375–82.

  34. 34.

    Brown TA, Jackson BA, Bythell BJ, Stenson AC. Benefits of multidimensional fractionation for the study and characterization of natural organic matter. J Chromatogr A. 2016;1470:84–96.

  35. 35.

    Kostyukevich Y, Kononikhin A, Popov I, Kharybin O, Perminova I, Konstantinov A, et al. Enumeration of labile hydrogens in natural organic matter by use of hydrogen/deuterium exchange Fourier transform ion cyclotron resonance mass spectrometry. Anal Chem. 2013;85:11007–13.

  36. 36.

    Gonsior M, Zwartjes M, Cooper WJ, Song W, Ishida KP, Tseng LY, et al. Molecular characterization of effluent organic matter identified by ultrahigh resolution mass spectrometry. Water Res. 2011;45:2943–53.

  37. 37.

    Dittmar T, Koch B, Hertkorn N, Kattner G. A simple and efficient method for the solid-phase extraction of dissolved organic matter (SPE-DOM) from seawater. Limnol Oceanogr Methods. 2008;6:230–5. https://doi.org/10.4319/lom.2008.6.230.

  38. 38.

    Li Y, Harir M, Lucio M, Kanawati B, Smirnov K, Flerus R, et al. Proposed guidelines for solid phase extraction of Suwannee River dissolved organic matter. Anal Chem. 2016;88:6680–8.

  39. 39.

    Perdue EM, Green NW. Isobaric molecular formulae of C, H, and O—a view from the negative quadrants of van Krevelen space. Anal Chem. 2015;87:5079–85.

  40. 40.

    Green NW, Perdue EM. Fast graphically inspired algorithm for assignment of molecular formulae in ultrahigh resolution mass spectrometry. Anal Chem. 2015;87:5086–94.

  41. 41.

    Zepp RG, Sheldon WM, Moran MA. Dissolved organic fluorophores in southeastern US coastal waters: correction method for eliminating Rayleigh and Raman scattering peaks in excitation–emission matrices. Mar Chem. 2004;89:15–36.

  42. 42.

    Rostad CE, Leenheer J a. Factors that affect molecular weight distribution of Suwannee river fulvic acid as determined by electrospray ionization/mass spectrometry. Anal Chim Acta. 2004;523:269–78.

  43. 43.

    Hockaday WC, Purcell JM, Marshall AG, Baldock JA, Hatcher PG. Electrospray and photoionization mass spectrometry for the characterization of organic matter in natural waters: a qualitative assessment. Limnol Oceanogr. 2009;7:81–95.

  44. 44.

    These A, Reemtsma T. Limitations of electrospray ionization of fulvic and humic acids as visible from size exclusion chromatography with organic carbon and mass spectrometric detection. Anal Chem. 2003;75:6275–81.

  45. 45.

    Herzsprung P, Hertkorn N, Von Tümpling W, Harir M, Friese K, Schmitt-kopplin P. Molecular formula assignment for dissolved organic matter (DOM) using high-field FT-ICR-MS: chemical perspective and validation of sulphur-rich organic components (CHOS) in pit lake samples. Anal Bioanal Chem. 2016;408:2461–9.

  46. 46.

    Kellerman AM, Dittmar T, Kothawala DN, Tranvik LJ. Chemodiversity of dissolved organic matter in lakes driven by climate and hydrology. Nat Commun. 2014;5:1–8.

  47. 47.

    D’Andrilli J, Foreman CM, Marshall AG, McKnight DM. Characterization of IHSS pony lake fulvic acid dissolved organic matter by electrospray ionization fourier transform ion cyclotron resonance mass spectrometry and fluorescence spectroscopy. Org Geochem. 2013;65:19–28.

  48. 48.

    Schendorf TM, Del Vecchio R, Koech K, Blough NV. A standard protocol for NaBH4 reduction of CDOM and HS. Limnol Oceanogr Methods. 2016;14:414–23.

  49. 49.

    Phillips SM, Smith D. Light absorption by charge transfer complexes in brown carbon aerosols. Environ Sci Technol Lett. 2014;1:382–6. https://doi.org/10.1021/ez500263j.

  50. 50.

    Phillips SM, Smith D. Further evidence for charge transfer complexes in brown carbon aerosols from excitation−emission matrix fluorescence spectroscopy. J Phys Chem A. 2015;119:4545–51.

  51. 51.

    Cartisano CM, Del Vecchio R, Bianca MR, Blough NV. Investigating the sources and structure of chromophoric dissolved organic matter (CDOM) in the North Pacific Ocean (NPO) utilizing optical spectroscopy combined with solid phase extraction and borohydride reduction. Mar Chem. 2018;204:20–35.

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We thank Mourad Harir for the assistance with mass spectrometry data acquisition.


This work was supported by the National Science Foundation grant (OCE-1357411) awarded to NVB and RDV.

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Correspondence to Neil V. Blough.

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Rossana Del Vecchio regrettably passed away on July 4, 2019.

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Bianca, M.R., Baluha, D.R., Gonsior, M. et al. Contribution of ketone/aldehyde-containing compounds to the composition and optical properties of Suwannee River fulvic acid revealed by ultrahigh resolution mass spectrometry and deuterium labeling. Anal Bioanal Chem 412, 1441–1451 (2020). https://doi.org/10.1007/s00216-019-02377-x

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  • ESI
  • Natural dissolved organic matter
  • Humic substances
  • Deuterium labeling
  • Chemical reduction