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A Step-Forward Method of Quantitative Analysis of Enzymatically Produced Isomaltooligosaccharide Preparations by AEC-PAD

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Abstract

In this paper, anion exchange chromatography coupled with pulsed amperometric detection has been successfully applied for the fine analysis of isomaltooligosaccharides (IMO) syrups where previous reported methods suffered from a lack of homologue oligosaccharides resolution. These syrups are made of a very complex mixture of glucose oligosaccharides characterized at the same time by their DP value (from 2 to ~15) and linkage types [α-(1–2, 3 or 6) and non-IMO α-(1–4)] and position. A mix of available commercial standards (17 species) was completely separated on a CarboPac PA-100 column at a flow rate of 1 mL min−1 and with a gradient of sodium acetate in 100 mM sodium hydroxide. The method was validated according to calibration curve, precision, recovery tests, limits of detection and quantitation. Calibration curves presented correlation coefficients greater than 0.98. The analytical method has been applied on real syrups, keeping a high performance separation of structurally close molecules and giving, for six determinations, very low relative SD for the available standard molecules (0.3–5.8%). The accuracy of the proposed method was tested by recovery measurements: first by spiking maltose on three different syrups and then by spiking six different sugar standards (20, 50 and 75% of the initial content) on a single syrup. Good recovery results (respectively, 96.5–99.7 and 97.1–102.7%) were found. The method was found sensible with limits of detection (signal-to-noise ratio of 3) between 0.048 and 0.124 μg mL−1 and limits of quantification (signal-to-noise ratio of 10) between 0.159 and 0.412 μg mL−1.

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Acknowledgments

This work was jointly supported by the Walloon Region and the company Meurens Natural throughout the FIRST DEI IMOBIOSE research project. We would like to thank Professor Eckhard Loos for providing us the kojioligosaccharides.

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Authors and Affiliations

  1. Unité de Chemie Biologique Industrielle, Faculté Universitaire des Sciences Agronomiques de Gembloux, Passage des Déportés 2, 5030, Gembloux, Belgium

    Dorothée Goffin, Christelle Robert, Bernard Wathelet & Michel Paquot

  2. Unité de Technologie des Industries Agro-alimentaires, Faculté Universitaire des Sciences Agronomiques de Gembloux, Passage des Déportés 2, 5030, Gembloux, Belgium

    Dorothée Goffin & Christophe Blecker

  3. Meurens Natural, Rue des martyrs, 21, 4650, Herve, Belgium

    Yves Malmendier

Authors
  1. Dorothée Goffin
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  2. Christelle Robert
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  3. Bernard Wathelet
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  4. Christophe Blecker
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  5. Yves Malmendier
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  6. Michel Paquot
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Correspondence to Dorothée Goffin.

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Goffin, D., Robert, C., Wathelet, B. et al. A Step-Forward Method of Quantitative Analysis of Enzymatically Produced Isomaltooligosaccharide Preparations by AEC-PAD. Chroma 69, 287–293 (2009). https://doi.org/10.1365/s10337-008-0875-0

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  • DOI: https://doi.org/10.1365/s10337-008-0875-0

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