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

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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.

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

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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.

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Author notes

  1. Matthias Mechelke and Jonathan Herlet contributed equally to this work.

Authors and Affiliations

  1. Department of Microbiology, TUM School of Life Sciences Weihenstephan, Technical University of Munich, Emil-Ramann-Str. 4, 85354, Freising, Germany

    Matthias Mechelke, Jonathan Herlet, Wolfgang H. Schwarz, Vladimir V. Zverlov, Wolfgang Liebl & Petra Kornberger

  2. Professorship for Wood Bioprocesses, TUM School of Life Sciences Weihenstephan, Technical University of Munich, Hans-Carl-von-Carlowitz-Platz 2, 85354, Freising, Germany

    J. Philipp Benz

  3. Institute of Molecular Genetics, Russian Academy of Science, Kurchatov Sq. 2, 123182, Moscow, Russia

    Vladimir V. Zverlov

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  1. Matthias Mechelke
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  2. Jonathan Herlet
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  3. J. Philipp Benz
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Correspondence to Petra Kornberger.

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Mechelke, M., Herlet, J., Benz, J.P. et al. HPAEC-PAD for oligosaccharide analysis—novel insights into analyte sensitivity and response stability. Anal Bioanal Chem 409, 7169–7181 (2017). https://doi.org/10.1007/s00216-017-0678-y

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