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Analytical and Bioanalytical Chemistry

, Volume 409, Issue 30, pp 7169–7181 | Cite as

HPAEC-PAD for oligosaccharide analysis—novel insights into analyte sensitivity and response stability

  • Matthias Mechelke
  • Jonathan Herlet
  • J. Philipp Benz
  • Wolfgang H. Schwarz
  • Vladimir V. Zverlov
  • Wolfgang Liebl
  • Petra KornbergerEmail author
Research Paper

Abstract

The rising importance of accurately detecting oligosaccharides in biomass hydrolyzates or as ingredients in food, such as in beverages and infant milk products, demands for the availability of tools to sensitively analyze the broad range of available oligosaccharides. Over the last decades, HPAEC-PAD has been developed into one of the major technologies for this task and represents a popular alternative to state-of-the-art LC-MS oligosaccharide analysis. This work presents the first comprehensive study which gives an overview of the separation of 38 analytes as well as enzymatic hydrolyzates of six different polysaccharides focusing on oligosaccharides. The high sensitivity of the PAD comes at cost of its stability due to recession of the gold electrode. By an in-depth analysis of the sensitivity drop over time for 35 analytes, including xylo- (XOS), arabinoxylo- (AXOS), laminari- (LOS), manno- (MOS), glucomanno- (GMOS), and cellooligosaccharides (COS), we developed an analyte-specific one-phase decay model for this effect over time. Using this model resulted in significantly improved data normalization when using an internal standard. Our results thereby allow a quantification approach which takes the inevitable and analyte-specific PAD response drop into account.

Graphical abstract

HPAEC-PAD analysis of oligosaccharides and determination of PAD response drop leading to an improved data normalization

Keywords

HPAEC-PAD Oligosaccharide analysis XOS AXOS PAD response factor Data normalization 

Notes

Acknowledgments

Financial support from the European Commission (Collaborative FP7-KBBE Project Valor Plus, contract No. 613802) and the German Federal Ministry of Education and Research (grant number FKZ: 031A556) is gratefully acknowledged.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Supplementary material

216_2017_678_MOESM1_ESM.pdf (2 mb)
ESM 1 (PDF 2031 kb)

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Copyright information

© Springer-Verlag GmbH Germany 2017

Authors and Affiliations

  1. 1.Department of Microbiology, TUM School of Life Sciences WeihenstephanTechnical University of MunichFreisingGermany
  2. 2.Professorship for Wood Bioprocesses, TUM School of Life Sciences WeihenstephanTechnical University of MunichFreisingGermany
  3. 3.Institute of Molecular GeneticsRussian Academy of ScienceMoscowRussia

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