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Sample Preparation and Analysis of Tomato Pectin Monomers

  • Rune SlimestadEmail author
  • Vilde Holm
  • Hilde Barsett
Short Communication
  • 4 Downloads

Abstract

A protocol for pectin monomer analysis is reported. It consists of hydrolysis, derivatization with 1-phenyl-3-methyl-5-pyrazolone (PMP) followed by UHPLC-analysis. A combination of enzymatic and acidic hydrolysis improved the yield of released aldoses. The tagging reaction should run for at least 1 h at 80 °C, thus longer than previously reported. Products were analysed by the use of a resistant C18-column and a gradient of acetonitrile in a volatile buffer (20 mM NH4HCO3/NH3, pH 9.2). Chromatograms revealed baseline separation of two pentoses, two deoxy-hexoses, three hexoses and two uronic acids within a 15 min run. Quantification was based on absorbance at 250 nm, whereas peak identities were confirmed by use of ESI–MS. The protocol was applied to pericarp fractions of tomato pectin, and the results were compared to those obtained from an established method based on GC-analysis. The new method had the advantage of being more rapid and produced simpler chromatograms.

Graphical Abstract

Keywords

UHPLC Pectin monomers PMP-tagging Tomato Pericarp fractions 

Notes

Funding

This study is part of the research project ‘Biofresh’ which is funded by the Research Council of Norway (project no 255613/E50).

Compliance with Ethical Standards

Conflict of Interest

The authors declare that there are no conflicts of interest.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.

References

  1. 1.
    O’Neill M, Albersheim P, Darvill A (1990) In: Dey PM, Harborne JB (eds) Methods in plant biochemistry vol 2 Carbohydrates. Academic Press, London, p 416Google Scholar
  2. 2.
    Harding SE, Tombs MP, Adams GG, Smestad Paulsen B, Tvete Inngjerdingen K, Barsett H (2017) An introduction to polysaccharide biotechnology, 2nd edn. CRC Press, Boca RatonCrossRefGoogle Scholar
  3. 3.
    Honda S, Akao E, Suzuki S, Okuda M, Kakehi K, Nakamura J (1989) Anal Biochem 180:351–357CrossRefGoogle Scholar
  4. 4.
    Xu G, Amicucci MJ, Cheng Z, Galermo AG, Lebrilla CB (2017) Analyst 143:200–207CrossRefGoogle Scholar
  5. 5.
    Dolan JA (2005) Guide to HPLC and LC-MS buffer selection. https://www.hplc.eu/Downloads/ACE_Guide_BufferSelection.pdf. Accessed 12 Apr 2018
  6. 6.
    Kirkland JJ, van Straten MA, Claessens HA (1995) J Chrom A 691:3–19CrossRefGoogle Scholar
  7. 7.
    Houben K, Jolie RP, Fraeye I, Van Loey AM, Hendrickx ME (2011) Carbohydr Res 346:1105–1111CrossRefGoogle Scholar
  8. 8.
    Arnous A, Meyer AS (2008) Food Bioprod Process 86:79–86CrossRefGoogle Scholar
  9. 9.
    Barsett H, Paulsen BS, Habte Y (1992) Carbohydr Poly 18:125–130CrossRefGoogle Scholar
  10. 10.
    Chambers RE, Clamp JR (1971) Biochem J 125:1009–1018CrossRefGoogle Scholar
  11. 11.
    Sweeley CC, Bentley R, Makita M, Wells WW (1963) J Am Chem Soc 85:2497–2507CrossRefGoogle Scholar
  12. 12.
    Ai Y, Yu Z, Chen Y, Zhu X, Ai Z, Liu S, Ni D (2016) J Chem 2016:6065813CrossRefGoogle Scholar
  13. 13.
    Wang HX, Zhao J, Li DM, Song S, Fu YH, Zhang LP (2015) Carbohydr Anal 402:95–101CrossRefGoogle Scholar
  14. 14.
    Yang X, Zhao Y, Wang Q, Wang H, Mei Q (2005) Anal Sci 21:1177–1180CrossRefGoogle Scholar
  15. 15.
    Claessens HA, van Straten MA, Kirkland JJ (1996) J Chrom A 728:259–270CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.PlantChem ASEikenNorway
  2. 2.Department of Pharmaceutical Chemistry, School of PharmacyUniversity of OsloOsloNorway

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