Excitation−emission matrix (EEM) fluorescence spectroscopy has been widely used to characterize dissolved organic matter (DOM) in water and soil. However, interpreting the >10,000 wavelength-dependent fluorescence intensity data points represented in EEMs has posed a significant challenge. Fluorescence regional integration, a quantitative technique that integrates the volume beneath an EEM, was developed to analyze EEMs. EEMs were delineated into five excitation−emission regions based on fluorescence of model compounds, DOM fractions, and marine waters or freshwaters. Volumetric integration under the EEM within each region, normalized to the projected excitation−emission area within that region and dissolved organic carbon concentration, resulted in a normalized region-specific EEM volume (Φi,n). Solid-state carbon nuclear magnetic resonance (13C NMR), Fourier transform infrared (FTIR) analysis, ultraviolet−visible absorption spectra, and EEMs were obtained for standard Suwannee River fulvic acid and 15 h...

Fluorescence excitation-emission matrix regional integration to quantify spectra for dissolved organic matter.

The polymerase chain reaction (PCR) is increasingly used as the standard method for detection and characterization of microorganisms and genetic markers in a variety of sample types. However, the method is prone to inhibiting substances, which may be present in the analysed sample and which may affect the sensitivity of the assay or even lead to false-negative results. The PCR inhibitors represent a diverse group of substances with different properties and mechanisms of action. Some of them are predominantly found in specific types of samples thus necessitating matrix-specific protocols for preparation of nucleic acids before PCR. A variety of protocols have been developed to remove the PCR inhibitors. This review focuses on the general properties of PCR inhibitors and their occurrence in specific matrices. Strategies for their removal from the sample and for quality control by assessing their influence on the individual PCR test are presented and discussed.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1365-2672.2012.05384.x

PCR inhibitors - occurrence, properties and removal.

The biochemical composition of dissolved organic matter (DOM) strongly influences its biogeochemical role in freshwater ecosystems, yet DOM composition measurements are not routinely incorporated into ecological studies. To date, the majority of studies of freshwater ecosystems have relied on bulk analyses of dissolved organic carbon and nitrogen to obtain information about DOM cycling. The problem with this approach is that the biogeochemical significance of DOM can only partially be elucidated using bulk analyses alone because bulk measures cannot detect most carbon and nitrogen transformations. Advances in fluorescence spectroscopy provide an alternative to traditional approaches for characterizing aquatic DOM, and allow for the rapid and precise characterization of DOM necessary to more comprehensively trace DOM dynamics. It is within this context that we discuss the use of fluorescence spectroscopy to provide a novel approach to tackling a longstanding problem: understanding the dynamics and biogeochemical role of DOM. We highlight the utility of fluorescence characterization of DOM and provide examples of the potential range of applications for incorporating DOM fluorescence into ecological studies in the hope that this rapidly evolving technique will further our understanding of the biogeochemical role of DOM in freshwater ecosystems.

https://aslopubs.onlinelibrary.wiley.com/doi/pdf/10.4319/lo.2010.55.6.2452

Fluorescence spectroscopy opens new windows into dissolved organic matter dynamics in freshwater ecosystems: A review

Dissolved organic matter (DOM) in aquatic systems originates from a range of sources. Some is allochthonous, transported from the surrounding landscape to the water body, and is derived from and influenced by the geology, land use and hydrology of its origin. Some is created in situ through microbial activity, which may provide an independent source of organic matter, or a recycling mechanism for that which has been transported into the water body. The relative contribution of each source depends upon the location and environmental conditions within and without the water body. Human activity is also a source of DOM, much of which is believed to be labile, which can enter the aquatic system through direct point discharges, diffuse leaching and aerial dispersal. Fluorescence spectroscopy can provide an excellent tool to source DOM fractions, and to monitor and understand DOM transformations in aquatic systems, as much DOM has an intrinsic fluorescence. In particular, recent advances in optical technology, enabling rapid investigation of shorter wavelengths, have enabled more detailed characterization of organic material and its reactions in water. In this article, we review the use of fluorescence spectroscopic techniques to measure the intrinsic fluorescence of organic matter and the application of fluorescent DOM analysis in marine waters, freshwaters and wastewaters. Copyright © 2007 John Wiley & Sons, Ltd.

https://onlinelibrary.wiley.com/doi/pdf/10.1002/rra.1005

Fluorescence analysis of dissolved organic matter in natural, waste and polluted waters—a review

Fluorescence as a potential monitoring tool for recycled water systems: a review.

Fluorescence measurements (emission scan, synchronous scan, and excitation-emission matrix [EEM] scan) were used to compare characteristics of two sources of dissolved organic carbon (DOC) from distinctly different origins: (i) a standard fulvic acid from the Suwannee River (SRF sample) and (ii) an unfractionated DOC sample from a tertiary wastewater treatment plant (MWW sample). Two methods were demonstrated that quantitatively differentiated allochthonous DOC (e.g., SRF) from autochthonous DOC (e.g., MWW). The MWW sample exhibited fluorescence peaks undetected in the SRF sample, at shorter wavelength pairs (e.g., 220 nm:300 to 350 nm) than the dominant peaks in the SRF sample (e.g., 220 nm:450 nm). These peaks may be associated with base or neutral fractions, potentially enriched in organic nitrogen. Effects of DOC concentration and solution pH were discussed. A simple procedure was recommended (pH = 3; DOC = 1 mg/L; dilution with 0.01 M KCl) that minimizes the need to correct spectra for inner-filter absorbance effects. A method, using synchronous fluorescence, to estimate the percentage of DOC from different sources when mixed together was also presented. Further work to understand the structural properties of DOC that fluoresce in wastewater samples, especially at shorter EEM wavelength pairs, will enable water managers to better understand the influence of wastewater on DOC in receiving waters (e.g., rivers, lakes).

Wen Chen

Papers

Fluorescence excitation-emission matrix regional integration to quantify spectra for dissolved organic matter.

Fluorescence analysis of a standard fulvic acid and tertiary treated wastewater.