Hp 6890 chromatograph

The NMR spectra were recorded on a Bruker AV 300 (300 MHz) or Bruker AV 400 (400 MHz) spectrometer in CDCl₃. The chemical shifts reported are relative to the center of the solvent resonance. IR spectra were recorded on a Bruker Alpha FT‐IR spectrometer using the ATR technique. GC was performed on a Hewlett–Packard HP 6890 chromatograph (HP5 column, 30 m, temperature program 50‐8‐260/5‐8‐280/5‐8‐300/5). Mass spectra of products were determined on an Agilent 6890/5973 GC‐MS. Elemental analyses were performed using a Leco TruSpec Micro CHNS elemental analyzer.

All the samples were prepared in a recirculating JACOMEX inert atmosphere (Ar) drybox and vacuum Schlenk lines. Glassware was dried overnight at 120 °C before use. NMR spectra were obtained using a Bruker DPX 400 MHz spectrometer (Bruker, Billerica, MA, USA). Chemical shifts for ¹H-NMR spectra were referenced to solvent impurities (herein, THF). NMR tubes were equipped with a J. Young valves were used for all ¹H-NMR experiments and catalytic tests. The GC-MS analysis was performed using n-decane as an internal standard. The reaction media aliquots were quenched by the addition of distilled water under air. The organic products were extracted using DCM, and injected into the GC-MS. Mass spectra were recorded on a Hewlett-Packart HP 5973 mass spectrometer (Hewlett Packard, Palo Alto, CA, USA) via a GC-MS coupling with a Hewlett-Packart HP 6890 chromatograph (Hewlett Packard, Palo Alto, CA, USA) equipped with a capillary column HP-5MS (50 m × 0.25 mm × 0.25 µm, Hewlett Packard, Palo Alto, CA, USA). Ionisation was due to an electronic impact (EI, 70 eV).

scr-MDA and sh-MDA cells were grown in 100 mm plates in routine medium or in routine medium supplemented with 50 μM AA as the sodium salt (Sigma) for 72 h. Cells were then washed 3 times with 0.1% BSA in ice-cold PBS, and scraped in ice-cold methanol and H₂O. Total lipids were extracted [17 (link)] and separated by TLC on silica-gel G60 plates (Merck) with hexane-ethylether-acetic acid (80:20:1; v/v/v) as the mobile phase for neutral lipid separation. All samples were chromatographed in parallel with pure lipid standards. To analyze the FA composition, total PL and TAG fractions were scraped from the plate and eluted with hexane: chloroform: methanol (3:2:1, v/v/v). FA methyl esters were obtained by reaction with BF₃ in methanol and analyzed by gas liquid chromatography in a Hewlett-Packard HP 6890 chromatograph equipped with an Omega Wax capillary column [2 (link)].