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From the first issue of 2016, Chromatography has changed its name to Separations.

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Chromatography 2015, 2(2), 277-292; doi:10.3390/chromatography2020277

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

Section for Plant Biochemistry, Department of Plant and Environmental Sciences, University of Copenhagen Thorvaldsensvej 40, 1871 Frederiksberg C, Denmark
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Academic Editor: Mary Boyce
Received: 15 April 2015 / Revised: 23 May 2015 / Accepted: 3 June 2015 / Published: 11 June 2015
(This article belongs to the Special Issue Solid Phase Micro-Extraction)
View Full-Text   |   Download PDF [1564 KB, uploaded 11 June 2015]   |  

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. View Full-Text
Keywords: HS-SPME; sesquiterpenoids; sesquiterpene synthases; gas chromatography; heat rearrangement; acid rearrangement; SPME fibers HS-SPME; sesquiterpenoids; sesquiterpene synthases; gas chromatography; heat rearrangement; acid rearrangement; SPME fibers
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Andersen, T.B.; Cozzi, F.; Simonsen, H.T. Optimization of Biochemical Screening Methods for Volatile and Unstable Sesquiterpenoids Using HS-SPME-GC-MS. Chromatography 2015, 2, 277-292.

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