Plasma and brain samples from non-Tg and SOD1G93A mice were obtained at ages (i) 60 days (before disease onset); (ii) 90 days (onset of disease); and (iii) 140 days (end stage, sacrifice) (Ludolph et al., 2010 (link)).
Analysis and quantitation of lipids in both plasma and brain were performed by ESI LC–MS/MS. For this purpose, samples were analyzed by direct infusion in a Q-TRAP4500 (ABSciex, Framingham, MA, United States) or coupled to a chromatographic separation in an Agilent 1260 HPLC. For chromatographic purposes, lipids were separated on a RP-C18 column (5 μm, 2 mm × 100 mm, Phenomenex Luna). The elution gradient consisted of solvent A: 0.05% acetic acid and solvent B: acetonitrile, 0.05% acetic acid with the following gradient at a flux of 700 μL/min: 0–0.2 min 30% B; 0.2–10 min 100% B; 10–11 min 100% B; 11–11.1 min 30% B; 11.1–15 min 30% B. The column was maintained during the run at a temperature of 30°C (Morgan et al., 2010 (link); Thomas et al., 2010 (link); Trostchansky et al., 2011 (link); Bonilla et al., 2013 (link)). Results were processed using Peak View software (ABSciex, Framingham, MA, United States). ESI-MS/MS was performed using an electrospray voltage set at 5 kV, and capillary temperature of 500°C.
Analysis and quantitation of lipids in both plasma and brain were performed by ESI LC–MS/MS. For this purpose, samples were analyzed by direct infusion in a Q-TRAP4500 (ABSciex, Framingham, MA, United States) or coupled to a chromatographic separation in an Agilent 1260 HPLC. For chromatographic purposes, lipids were separated on a RP-C18 column (5 μm, 2 mm × 100 mm, Phenomenex Luna). The elution gradient consisted of solvent A: 0.05% acetic acid and solvent B: acetonitrile, 0.05% acetic acid with the following gradient at a flux of 700 μL/min: 0–0.2 min 30% B; 0.2–10 min 100% B; 10–11 min 100% B; 11–11.1 min 30% B; 11.1–15 min 30% B. The column was maintained during the run at a temperature of 30°C (Morgan et al., 2010 (link); Thomas et al., 2010 (link); Trostchansky et al., 2011 (link); Bonilla et al., 2013 (link)). Results were processed using Peak View software (ABSciex, Framingham, MA, United States). ESI-MS/MS was performed using an electrospray voltage set at 5 kV, and capillary temperature of 500°C.
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