Optimization of Biochemical Screening Methods for Volatile and Unstable Sesquiterpenoids Using HS-SPME-GC-MS

Trine Andersen, Federico Cozzi, Henrik Toft Simonsen

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Abstract

HS-SPME-GC-MS has been suggested as a fast and robust analytical platform for the product characterization of sesquiterpene synthases. The choice of fiber and injection temperature can have a significant effect on the observed product profile, due to the chemical rearrangements that can occur on the fiber material. Here we present a systematic study on the effects of fiber choice and injection port temperature on the observed sesquiterpenoid profile of four sesquiterpene synthases expressed in Nicotiana benthamiana. We found that the absorbent material PDMS was much less likely to support acid-induced rearrangement of sesquiterpenoids when compared to the adsorbent materials PDMS/DVB, PDMS/CAR, and PDMS/CAR/DVB. Furthermore, utilizing an injection port temperature at 160 °C almost eliminated the inherent thermal instability of germacrene sesquiterpenoids. Thus, for fast screening of sesquiterpene synthases, the results suggest that PDMS fibers and an injection temperature of 160 °C provide a fast and reproducible HS-SPME GC-MS method when using H2 as carrier gas.
Original languageEnglish
JournalChromatography
Volume2
Issue number2
Pages (from-to)277-292
Number of pages16
ISSN1342-8284
DOIs
Publication statusPublished - 2015
Externally publishedYes

Bibliographical note

This is an open access article distributed under the Creative Commons Attribution License (CC BY 4.0).

Keywords

  • HS-SPME
  • Sesquiterpenoids
  • Sesquiterpene synthases
  • Gas chromatography
  • Heat rearrangement
  • Acid rearrangement
  • SPME fibers

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