The triacylglycerol (TAG) composition of coffee beans (Coffea canephora P.) was determined by reversed phase liquid chromatography-mass spectrometry and tandem mass spectrometry. The TAGs were separated on a Microsorb RP C-18 column in series with a Supelcosil RP C-18 column using isocratic elution with acetonitrile/2-propanol/hexane (v/v/v, 57:38:5) as the mobile phase at a flow rate of 1 mL/min for 100 min. Under these conditions, 13 TAGs were identified (elution order): trilinoleyl-glycerol (LLL, 11.76%), dilinolenoyl-palmitoyl-glycerol (PLnLn, 2.94%), dilinoleyl-oleyl-glycerol (OLL, 7.77%), dilinoleyl-palmitoyl-glycerol (PLL, 25.90%), dipalmitoyl-linolenoyl-glycerol (PPLn, 1.66%), dioleoyl-linoleyl-glycerol (OOL, 1.68%), dilinoleyl-stearyl-glycerol (SLL, 8.28%), palmitoyl-oleyl-linoleyl-glycerol (POL, 8.76%), dipalmitoyl-linoleyl-glycerol (PPL, 13.74%), dilinoleyl-arachidyl-glycerol (ALL, 3.51%), trioleoyl-glycerol (OOO, 2.33%), palmitoyl -linoleyl-stearyl -glycerol (PLS, 8.73%), and distearoyl-linolenonyl-glycerol (SSLn, 2.91%). The relative composition (%) was obtained directly from the data system with the photodiode array detector.