, Volume 81, Issue 2, pp 203–213 | Cite as

Fractionation of Dissolved Organic Matter on Coupled Reversed-Phase Monolithic Columns and Characterisation Using Reversed-Phase Liquid Chromatography-High Resolution Mass Spectrometry

  • Sara Sandron
  • Noel W. Davies
  • Richard Wilson
  • Alfonso Rojas Cardona
  • Paul R. Haddad
  • Pavel N. Nesterenko
  • Brett Paull
Part of the following topical collections:
  1. 2nd ACROSS International Symposium on Advances in Separation Science (ASASS 2016)


Eleven Onyx monolithic C18 columns (100 x 3 mm) were connected in series to obtain a high-capacity reversed-phase HPLC column providing 110,000 theoretical plates. The column was used to fractionate a complex mixture of semi-polar and apolar components within marine dissolved organic matter (DOM), isolated using solid-phase extraction (SPE) on poly(styrene–divinylbenzene) cartridges, and Suwannee River natural organic matter, isolated by reverse osmosis. In each case, 15 isolated fractions were further separated using a second-dimension reversed-phase HPLC coupled with high-resolution mass spectrometry. Successful fractionation of the major compositional materials within DOM (i.e. carboxylic-rich alicyclic molecules, CRAM) in order of decreasing polarity was confirmed. Upon formulae assignment, homologous series of compounds were fractionated according to decreasing O/C ratio and increasing H/C ratio. Different compounds with the same exact masses were separated across six fractions, highlighting the immensely complex nature of the material, with the presence of potentially hundreds of molecules for each molecular formula. Both samples showed clear compositional differences not only related to source (seawater or freshwater), but also to the extraction method applied (SPE or reverse osmosis).


Dissolved organic matter Fractionation Coupled monolithic columns Reversed-phase liquid chromatography High-resolution mass spectrometry 



This research was supported under Australian Research Council’s Discovery Projects funding scheme (Project Number DP130101518).

Compliance with Ethical Standards

Conflict of interest

The authors declare no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

10337_2017_3324_MOESM1_ESM.docx (3.2 mb)
Supplementary material 1 (DOCX 3227 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  1. 1.Australian Centre for Research on Separation Sciences, School of Physical SciencesUniversity of TasmaniaHobartAustralia
  2. 2.Central Science LaboratoryUniversity of TasmaniaHobartAustralia

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