Polar Fraction Analysis of Metabolites

Polar fraction analysis were carried out following Valledor, et al. [84 (link)] protocol with some minor changes on a triple quad instrument (TSQ Quantum GC; Thermo). In brief, 1 µL of sample was injected, and GC separated into a HP-5MS capillary column (30 m 0.25 mm) (Agilent Technologies). Oven temperature was increased from 80 °C to 200 °C at a 3 °C per min rate and then reduced to 25 °C at 10 °C per min and maintained at 25 °C for 3 min, followed by 4 min of post-run conditions at 30 °C. Mass spectrometer was operated in electron impact (EI) mode at 70 eV in a scan range of m/z 40–600. The identification of metabolites was based on the spectral characteristics and GC retention times of each individual metabolite through its comparison with the retention times and spectral characteristics of standards available in our in-house library and in Golm Metabolome Database [86 (link)].

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Colina F., Carbó M., Meijón M., Cañal M.J, & Valledor L. (2020). Low UV-C stress modulates Chlamydomonas reinhardtii biomass composition and oxidative stress response through proteomic and metabolomic changes involving novel signalers and effectors. Biotechnology for Biofuels, 13, 110.

Publication 2020

TSQ Quantum GC HP-5MS capillary column

Capillary Electron Library Metabolome Retention Scan

Corresponding Organization :

Other organizations : Universidad de Oviedo

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Variable analysis

independent variables

Sample injection volume (1 µL)

dependent variables

Metabolite identification based on spectral characteristics and GC retention times

control variables

GC column (HP-5MS capillary column, 30 m 0.25 mm)
Oven temperature program (increased from 80 °C to 200 °C at 3 °C/min, decreased to 25 °C at 10 °C/min, maintained at 25 °C for 3 min, and post-run conditions at 30 °C for 4 min)
Mass spectrometer operating conditions (electron impact (EI) mode at 70 eV, scan range of m/z 40–600)

positive controls

Retention times and spectral characteristics of standards available in the in-house library and the Golm Metabolome Database

negative controls

Not explicitly mentioned

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