A simple guideline to apply excitation-emission matrix spectroscopy (EEMs) for the characterization of dissolved organic matter (DOM) in anoxic marine sediments

Marine sediments represent a major carbon reservoir on Earth. Dissolved organic matter (DOM) in pore waters accumulates products and intermediates of carbon cycling in sediments. The application of excitation-emission matrix spectroscopy (EEMs) in the analysis of subseafloor DOM samples is largely unexplored due to the redox-sensitive matrix of anoxic pore water. Therefore, this study aims to investigate the interference caused by the matrix on EEMs and propose a guideline to prepare pore water samples from anoxic marine sediments. The parameters determined by fluorescence spectra include 3D-index derived from EEMs after parallel factor analysis (PARAFAC), fluorescence index (FI) (contribution of terrigenous DOM), biological index (BIX) and humification index (HIX) derived from 2D emission spectra. First, we investigated the impacts of extensively-presented ions as typical electron acceptors, which are utilized by anaerobic microbes and stratified in marine sediments: Fe(II), Fe(III), Mn(II) and sulfide in anoxic pore water resulted in biases of fluorescent signals. We proposed threshold concentrations of these ions when the interference on EEMs occurred. Effective removal of sulfide from sulfide-rich samples could be achieved by flushing with N-2 for 2 min. Second, the tests based on DOM standard were further verified using pristine samples from marine sediments. There was a significant change in the fluorescence spectra of DOM in anoxic sediments from the Rhone Delta. This study demonstrated that the change was caused by oxidation of the matrix rather than the intrinsic alteration of DOM. It was confirmed by extracted DOM via both EEMs analysis and Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS). Slight oxidation of sulfur-containing compounds (e.g., sulfhydryl) and polyphenol-like compounds occurred. Finally, a sample preparation sequence is proposed for pore water from anoxic sediments. This method enables measurement with small volumes of the sample (e.g., 50 mu L in this study) and ensures reliable data without the interference of the redox-sensitive matrix. This study provides access to the rapid analysis of DOM composition in marine sediments and can potentially open a window into examining the carbon cycling of the marine deep biosphere.

Automated summary

Marine sediments serve as a major carbon reservoir and accumulate dissolved organic matter (DOM) in pore waters, which is essential for carbon cycling. This study investigates the interference caused by the matrix in analyzing subseafloor DOM samples using excitation-emission matrix spectroscopy (EEMs) and proposes a guideline for preparing pore water samples. The fluorescence spectra parameters, including 3D-index, fluorescence index (FI), biological index (BIX), and humification index (HIX), were analyzed to determine the impacts of different ions present in anoxic pore water. The study suggests threshold concentrations for these ions and a method for removing sulfide from sulfide-rich samples. The research findings also demonstrate that the change in fluorescence spectra of DOM in anoxic sediments is primarily caused by oxidation of the matrix rather than intrinsic alteration of DOM.