Two-stage bile preparation with acetone for recovery of fluorescent aromatic compounds (FACs)

► Type and volume of organic solvent affect recovery of FACs from fish bile. ► Significant FAC quantities may be lost in centrifugation pellets. ► Two-stage preparation with acetone is more efficient than previously applied methods. ► L-LEs may not be appropriate for the preparation of fish bile samples containing FACs. ► Proteins have a significant negative influence on the detection of biliary FACs.