Fatty acid analysis

Collect cultured cells(1*107 cells per sample) into centrifuge tubes. After grinding and centrifugation of the cells, the supernatant was taken and vortexed with chloroform, centrifuged, and the lower layer liquid was taken and placed in a sample bottle. It was concentrated and dried, and dissolved in methanol.The cellular metabolites were dissolved in methanol using ultra high performance liquid chromatography ion mobility quadrupole time-of-flight mass spectrometer (Acquity I-class; Waters). Cell samples include PANC-1 and Capan-2 cells upon SCD knockdown or TGM2 knockdown. The experiment was conducted by Instrumental Analysis Center of Shanghai Jiao Tong University. The mobile phase A: Containing 0.1% formic acid water; mobile phase B: Containing 0.1% formic acid acetonitrile (1/1). The gradient was as follows: 95%A and 5%B (initial); 80%A and 20%B (5 min) ; 100%B (18 min); 95%A and 5%B (28 min) with a flow rate of 0.4ml/min in column BEH C18 1.7um，2.1*100mm, column temperature at 45℃. MS conditions were follows: the acquisition mode at MSE; ionization mode at ESI negative, capillary voltage at 2 KV (negative), cone voltage at 40 V, desolvation temperature at 450℃, desolvation gas at 900 L/h, cone gas at 50L/h, source temperature at 115℃, acquisition range at 50 to 1000 m/z, scan rate at 0.2 s, collision energy at 6 eV/20～45 eV; lockmass at 250 pg/μL of leucine enkephalin; flow rate at 10 μL/min; interval at 0.5 s; sample time at 0.5 s; collision energy at 6 eV. Data acquisition and analysis were carried out by Waters UNIFI 1.9.4. A series of FA samples is standard.