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 Kornberger

Research output: Contribution to journalArticlepeer-review

32 Scopus citations

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. [Figure not available: see fulltext.].

Original languageEnglish
Pages (from-to)7169-7181
Number of pages13
JournalAnalytical and Bioanalytical Chemistry
Volume409
Issue number30
DOIs
StatePublished - 1 Dec 2017

Keywords

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

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