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标题:Chemical characterization methods for the analysis of structural extracellular polymeric substances (EPS).
时间:2020-03-26 20:19:24
DOI:10.1016/j.watres.2019.03.068
PMID:30953855
作者:Felz, Simon;Vermeulen, Pascalle;van Loosdrecht, Mark C. M
关键词:Extracellular polymeric substances (EPS);EPS analysis;Colorimetric methods;Biofilm;Aerobic granular sludge
出版源: Water Research ,2019
摘要:Biofilm structure and functionality depend on extracellular polymeric substance (EPS), but analytical methods for EPS often lack specificity which limits progress of biofilm research.EPS were extracted from aerobic granular sludge and analyzed with frequently applied colorimetric methods. The colorimetric methods were evaluated based on their applicability for EPS analysis. EPS fractions of interest were proteins, sugars, uronic acids and phenolic compounds. The applied methods (Lowry method, bicinchoninic acid assay, phenol sulfuric acid method, carbazole sulfuric acid method) were investigated in terms of their sensitivity towards the selected standard compound. Interference of compounds present in EPS with the colorimetric methods was further evaluated. All methods showed to be highly depending on the choice of standard compound and susceptible towards interference by compounds present in EPS.This study shows that currently used colorimetric methods are not capable of accurately characterizing EPS. More advanced methods are needed to be able to draw conclusions about biofilm composition, structure and functionality. (C) 2019 The Authors. Published by Elsevier Ltd.
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目录:
  • Chemical characterization methods for the analysis of structural extracellular polymeric substances (EPS)
    • 1. Introduction
    • 2. Material and methods
      • 2.1. Sample collection
      • 2.2. Extraction of structural EPS from aerobic granular sludge
      • 2.3. Colorimetric assays
        • 2.3.1. Protein determination
          • 2.3.1.1. Lowry method
          • 2.3.1.2. BCA assay
        • 2.3.2. Saccharide determination with the phenol sulfuric acid method
        • 2.3.3. Uronic acid determination with the carbazole sulfuric acid method
        • 2.3.4. Phenolic compound determination with the modified Lowry method
      • 2.4. HPAEC-PAD analysis of monosaccharides in EPS
    • 3. Results
      • 3.1. Chemical characterization of EPS with colorimetric methods
      • 3.2. Possible cross-interferences of compounds present in EPS with the here applied colorimetric methods
      • 3.3. Monosaccharide analysis of EPS with HPAEC-PAD
    • 4. Discussion
      • 4.1. Selection of the standard
      • 4.2. Cross-interference in EPS analysis
    • 5. Conclusion
    • Declaration of interests
    • Author agreement
    • Acknowledgements
    • References

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