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

Chromatography

Trine Bundgaard Andersen¹  Federico Cozzi¹  Henrik Toft Simonsen² 

[1] Section for Plant Biochemistry, Department of Plant and Environmental Sciences, University of Copenhagen Thorvaldsensvej 40, 1871 Frederiksberg C, Denmark;

关键词: HS-SPME;    sesquiterpenoids;    sesquiterpene synthases;    gas chromatography;    heat rearrangement;    acid rearrangement;    SPME fibers;   

DOI  :  10.3390/chromatography2020277

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
© 2015 by the authors; licensee MDPI, Basel, Switzerland.