期刊
出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jchromb.2008.06.039
关键词
metabolite extraction; mass spectrometry; micro-liquid chromatography; plant metabolomics; pre-LC-MS treatment
This study treats the optimization of methods for homogenizing Arabidopsis thaliana plant leaves as well as cell cultures, and extracting their metabolites for metabolomics analysis by conventional liquid chromatography electrospray ionization mass spectrometry (LC-ESI/MS). Absolute recovery, process efficiency and procedure repeatability have been compared between different pre-LC-MS homogenization/extraction procedures through the use of samples fortified before extraction with a range of representative metabolites. Hereby, the magnitude of the matrix effect observed in the ensuing LC-MS based metabolomics analysis was evaluated. Based on relative recovery and repeatability of key metabolites, comprehensiveness of extraction (number of m/z-retention time pairs) and clean-up potential of the approach (minimum matrix effects), the most appropriate sample pre-treatment was adopted. It combines liquid nitrogen homogenization for plant leaves with thermomixer based extraction using MeOH/H(2)O 80/20. As such, an efficient and highly reproducible LC-MS plant metabolomics set-up is achieved, as illustrated by the obtained results for both LC-MS (8.88% +/- 5.16 versus 7.05% +/- 4.45) and technical variability (12.53% +/- 11.21 versus 9.31% +/- 6.65) data in a comparative investigation of A. thaliana plant leaves and cell cultures, respectively. (C) 2008 Elsevier B.V. All rights reserved.
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