Comparative study of chromatographic methods for the analysis of exopolysaccharides from archaeal biofilms
- 1Applied Analytical Chemistry (AAC), University of Duisburg-Essen, Essen, Germany
- 2Teaching and Research Center for Separation (TRC), University of Duisburg-Essen, Essen, Germany
- 3Molecular Enzyme Technology and Biochemistry (MEB), University of Duisburg-Essen, Essen, Germany
- 4Aquatic Microbiology, University of Duisburg-Essen, Essen, Germany
Microorganisms, such as archaea, favour life in a biofilm rather than the planktonic form of life. A biofilm is defined as a community of microorganisms embedded in a self-produced matrix of hydrated extracellular polymeric substances (EPS), mainly polysaccharides (PS), proteins and extracellular DNA. The polysaccharides form a three-dimensional network, which provides stability of the biofilm and mediates the adhesion to surfaces. [1] Analysis of the monomeric composition of PS requires chromatographic separation and identification by mass spectrometry (MS).
A comparative study of different chromatographic methods for the analysis of the monomeric composition of exopolysaccharides from archaeal biofilms from Sulfolobus acidocaldarius has been carried out. For this study, different chromatographic separation methods, such as supercritical fluid chromatography (SFC), hydrophilic interaction liquid chromatography (HILIC) reversed-phase liquid chromatography (RP-LC) and gas chromatography (GC), each coupled to mass spectrometry, were developed and compared by means of separation performance and sensitivity, using authentic standards.
The study revealed, that each method features distinct advantages and disadvantages over the other methods. For example, when using SFC-MS, no derivatization is necessary and soft ionization conditions can be used. [2] However, the HILIC-MS and RP-LC-MS methods show significantly greater separation performances for the analysis of the monosaccharide composition. [3] All investigated methods show similar quantification limits in the sub-mg/L range.
Finally, the developed chromatographic methods were applied to real biofilm samples of the thermoacidophilic archaeon Sulfolobus acidocaldarius. To determine the monomeric composition of the exopolysaccharides from these archaeal biofilms, the extracellular polymeric substances were extracted from the biofilm and then the PS were hydrolyzed.
Literature:
[1] H.-C. Flemming, Nat. Microbiol. Rev. 2010, 8, 623 - 633. [2] M. Lafosse, J. Chromatogr. A, 1996, 720, 61-73. [3] V. Sieber, J. Chromatogr. A, 2014, 1350, 44–50.
How to cite: Meyer, M., Kuschmierz, L., Siebers, B., Wingender, J., and Schmitz, O. J.: Comparative study of chromatographic methods for the analysis of exopolysaccharides from archaeal biofilms, biofilms 9 conference, Karlsruhe, Germany, 29 September–1 Oct 2020, biofilms9-58, https://doi.org/10.5194/biofilms9-58, 2020
