Pe 17 0 17 0

Lipids from liver samples were processed at the Montreal Heart Institute as previously described 72 .
Briefly, liver samples (40 mg) were all processed in one batch of extraction after spiking with the following lipid standards used to validate the reproducibility of the extraction and measurement:
LPC 13:0, PC14:0/14:0, PC 19:0/19:0, PE17:0/17:0, PS12:0/12:0 and PG15:0/15:0 (Avanti Polar Lipids Inc., Alabaster, USA). Lipids were extracted into a methyl tert-butyl ether (MTBE/methanol/H20) mixture, followed by the following steps: i) extraction with ethyl acetate, drying under vacuum, and iii) solubilization in methanol/chloroform by vortexing and sonication. The samples were diluted (1:10 in methanol/chloroform, 2:1) and the diluted samples Qualitative Analysis software package (version B.06, Agilent). MS features were first annotated using a public database (Metlin) by alignment with our in-house database containing more than 500 lipids which have been previously identified in human plasma by MSMS analysis for which spectra were manually interpreted, similar to Godzien et al 73 . The significant MS features were finally validated using MSMS analysis.