HPLC-ESI-MS Metabolite Identification Protocol

Samples collected in 2011 were extracted and analyzed by HPLC-ESI-MS as previously described^{26 (link)}. In brief, the extracts were analyzed by HPLC using a Beckman Coulter Gold 127 Solvent Module coupled to a Bruker Esquire 6000 ion trap mass spectrometer (Bruker Daltonik GmbH, Bremen, Germany) equipped with an electrospray ionization source. Negative ion spectra were recorded in the range 50–2000 m/z (full scan mode, 13,000 m/z per second). For metabolite identification, MS/MS and MS3 spectra were recorded in negative mode. Metabolites were putatively identified by comparing the m/z values, fragmentation patterns (MS/MS and MS3) and retention times of each signal with those of an in-house library of authentic standards. When commercial standards were not available, m/z and fragmentation patterns were compared with those published in the literature or on-line databases such as MassBank (www.massbank.jp/en/database.html) and Human Metabolome Database (http://www.hmdb.ca/).

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Zenoni S., Amato A., D’Incà E., Guzzo F, & Tornielli G.B. (2020). Rapid dehydration of grape berries dampens the post-ripening transcriptomic program and the metabolite profile evolution. Horticulture Research, 7, 141.

Publication 2020

Esquire 6000

Gold Hplc Human Library Metabolome Ms ms Retention Scan Solvent

Corresponding Organization : University of Verona

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Variable analysis

independent variables

Sample collection year (2011)

dependent variables

Metabolite identification based on m/z values, fragmentation patterns (MS/MS and MS3), and retention times

control variables

HPLC-ESI-MS analytical method as previously described in ref. 26
In-house library of authentic standards for metabolite identification
Comparison with literature and online databases (MassBank, Human Metabolome Database) when commercial standards were not available

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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