Oak bark is a rich niche for beneficial bioactive compounds. It is known that the amount of the compounds found in plant tissues can depend on species, genotype, growth site, etc., but it is unclear whether oak phenology, i.e., late or early bud burst, can also influence the amount of phenols and antioxidants that can be extracted. We tested two Quercus robur populations expressing different phenology and five half-sib families in each population to see how phenology, genotype, as well as extrahent differences (75% methanol or water) can determine the total phenol, total flavonoid content, as well as antioxidant activity. Significant statistical differences were found between half-sib families of the same population, between populations representing different oak phenology and different extrahents used. We determined that the extraction of flavonoids was more favorable when using water. So was antioxidant activity using one of the indicators, when significant differences between extrahents were observed. Furthermore, in families where there was a significant difference, phenols showed better results when using methanol. Overall, late bud burst families exhibited higher levels in all parameters tested. Thus, we recommend that for further bioactive compound extraction, all these factors be noted.