We used the same method as previously described [25 (link)]. In brief, total lipids were extracted from the RBC cell membranes using Folch’s method [30 (link)]. Samples were split into 3 parts. Fatty acid levels were measured with gas chromatography [31 (link)]. Phospholipids [32 (link),33 (link)] and sterols (cholesterol and cholestanol) [34 (link)] were measured with liquid chromatography tandem mass spectrometry (LC-MS/MS).
Lipids analysis was conducted at the Mass Spectroscopy Department at the Saint-Antoine Hospital, Paris, France. Internal standards for the determination of phospholipids, sterols, and sphingolipids are from Avanti Polar Lipids (Alabaster, AL, USA). Ammonium acetate and trimethylsilyl diazomethane came from Sigma-Aldrich (Saint Quentin Fallavier, France). Methanol, chloroform, n-hexane, heptane, and isopropanol of mass spectrometry quality were obtained from VWR (Fontenay Sous-Bois, France). Total lipids were extracted from the RBC cell membranes based on the methods of Folch et al. Samples were divided into 3 parts.
Lipids analysis was conducted at the Mass Spectroscopy Department at the Saint-Antoine Hospital, Paris, France. Internal standards for the determination of phospholipids, sterols, and sphingolipids are from Avanti Polar Lipids (Alabaster, AL, USA). Ammonium acetate and trimethylsilyl diazomethane came from Sigma-Aldrich (Saint Quentin Fallavier, France). Methanol, chloroform, n-hexane, heptane, and isopropanol of mass spectrometry quality were obtained from VWR (Fontenay Sous-Bois, France). Total lipids were extracted from the RBC cell membranes based on the methods of Folch et al. Samples were divided into 3 parts.
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