High-performance liquid chromatography with confirmation by UV-visible photodiode array detector-positive electrospray ionization-mass spectrometry [HPLC-UV-vis-DAD-(+ESI)-MS] with enhanced fragmentation by appropriate adjustment of the cone voltage was used to determine bioflavonoid content of five citrus species (tangerine, sanguine, sour orange, lemon and grapefruit) cultivated in Greece which come from citrus varieties analyzed for the first time. The main groups of bioflavonoids found in the juice of the citrus species according to HPLC retention times, spectral data and literature references were O-glycosylated flavanones and flavones, C-glucosylated flavones, O-glucosylated flavones, O-C-glucosylated flavones like saponarin and a phenolic derivative.