Mouse peritoneal exudates and human macrophages with supernatants were extracted for LC-MS/MS–based mediator analysis. We used a UPLC I-Class system (Waters, Milford, MA, USA) equipped with an AB Sciex Instruments 6500 Q-TRAP mass spectrometer (Applied Biosystems, Foster City, CA, USA). Acquisition was carried out in a negative ionization mode. The mobile phase consisted of methanol/water/acetic acid (60:40:0.01, v/v/v) and was ramped to 85:15:0.01 over 30 min and to 100:0:0.01 over the next 5 min at a flow rate of 200 ml/min. Instrument control and data acquisition were performed using Analyst 1.6 software (Applied Biosystems). A minimum of six diagnostic ions and retention time were used for lipid identification (23). Quantification was based on peak area of multiple reaction monitoring transitions and linear calibration curve of each compound.

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