The present study aims to predict the regiospecific glucuronidation of three dihydroxyflavones and seven monohydroxyflavones in human liver and intestinal microsomes using recombinant UGT isoforms. Seven monohydroxyflavones (or HFs), 2'-, 3'-, 4'-, 3-, 5-, 6-, and 7-hydroxyflavone, and three dihydroxyflavones (or diHFs), 3,7-dihydroxyflavone (3,7-diHF), 3,5-dihydroxyflavone (3,5-diHF), and 3,4'-dihydroxyflavone (3,4'-diHF), were chosen, and rates were measured at 2.5, 10, and 35 microM. The results indicated that the position of glucuronidation of three diHFs could be determined by using the UV spectra of relevant HFs. The results also indicated that UGT1A1, UGT1A7, UGT1A8, UGT1A9, UGT1A10 and UGT2B7 are the most important six UGT isoforms for metabolizing the chosen flavones. Regardless of isoforms used, 3-HF was always metabolized the fastest whereas 5-HF was usually metabolized the slowest, probably due to the formation of an intramolecular hydrogen bond between 4-carbonyl and 5-OH group. Relevant UGT isoform-specific metabolism rates generally correlated well with the rates of glucuronidation in human intestinal and liver microsomes at each of the three tested concentrations. In conclusion, the glucuronidation "fingerprint" of seven selected monohydroxyflavones was affected by UGT isoforms used, positions of the -OH group, and the substrate concentrations, and the rates of glucuronidation by important recombinant UGTs correlated well with those obtained using human liver and intestinal microsomes.