MRM results of remission and IBD patient blood

Metabolomics of free fatty acids

Freshly obtained intestine tissues were extracted with 80% methanol and stored at -80°C. Metabolomics analysis was carried out with MRM (Multiple Reaction Monitoring mode) mode of UPLC-QQQ-MS/MS to quantify free fatty acids via negative ionization. Samples were separated by an ACQUITY UPLC I-Class system (fixed loop), equipped with a CORTECS T3 2.7 mm (2.1*3.0 mm) analytical column. A sample of 2 mL was injected at a flow rate of 1.3 mL/min. Mobile phase A was 0.01% formic acid_(aq) containing 0.2 mM Ammonium Formate and mobile phase B was 50% isopropanol in acetonitrile containing 0.01% formic acid_(aq) containing 0.2 mM Ammonium Formate. The free fatty acids were eluted from the column and separated with a gradient of 50-98% Mobile phase B. The column was then re-euquilibrated to initial conditions. The analytical column temperature was maintained at 60 ^oC .

MS conditions

Multiple Reaction Monitoring (MRM) analyses were performed using a Xevo TQ-S micro Mass Spectrometer. All experiments were performed in negative electrospray ionization (ESI-) mode. The ion source temperature and capillary voltage were kept constant and set to 150 ^oC and 2.0 kV respectively. The cone gas flow rate was 50 L/hr and desolvation temperature was 650 ^oC.