Lpc 13 0

The reference standards of LPC(13:0) (internal standard, IS), LPC(14:0), LPC(16:0), LPC(17:0), LPC(18:1) and LPC(20:0) were purchased from Avanti Polar Lipids Inc. (Alabaster, AL, USA). Mass spectrometry grade-methanol and acetonitrile were obtained from Merck Inc. (Darmstadt, HE, Germany). Analytical reagent-chloroform and HPLC grade-formic acid were purchased from Xilong Technology Co., Ltd. (Shantou, China). Phosphate buffer saline (PBS) was supplied by Servicebio Technology Co., Ltd. (Wuhan, China). Bovine serum albumin (BSA) was provided by Energy Chemical Technology Co., Ltd. (Shanghai, China). CH and APAP were purchased from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan). Isoflurane was supplied by RWD Life Science CO., Ltd. (Shenzhen, China)

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.