BACKGROUND: Previously, we reported the presence of hydroxytyrosol in red wine and higher human urinary recovery of total hydroxytyrosol than that expected after a single red wine intake. We hypothesized that the alcohol present in wine could promote endogenous hydroxytyrosol generation. OBJECTIVE: The objective was to assess the relation between alcohol consumption and urinary hydroxytyrosol concentrations. DESIGN: This was a cross-sectional study with baseline data from a subsample of the PREvención con DIeta MEDiterránea (PREDIMED) trial, an intervention study directed at testing the efficacy of the Mediterranean diet on the primary prevention of cardiovascular disease. Participants included 1045 subjects, aged 55-80 y, who were at high cardiovascular risk. Alcohol consumption was estimated through a validated food-frequency questionnaire. Urinary hydroxytyrosol and ethyl glucuronide, a biomarker of alcohol consumption, were measured. RESULTS: Urinary ethyl glucuronide concentrations were directly related to alcohol and wine consumption (P < 0.001) as well as to urinary hydroxytyrosol in both sexes (P < 0.001). The degree of alcohol consumption was directly associated with urinary hydroxytyrosol in male alcohol consumers (P < 0.001). Multivariate logistic regression analyses showed a significant linear trend (P < 0.05) for elevated hydroxytyrosol concentrations with an increase in alcohol consumption. Intakes of >20 g (2 drinks)/d and >10 g (1 drink)/d alcohol in men and women, respectively, were associated (P < 0.05) with elevated concentrations of hydroxytyrosol. CONCLUSIONS: We report for the first time a direct association between urinary hydroxytyrosol and alcohol consumption at a population level. These findings reinforce previous work in human and animal models that examines wine as a source of hydroxytyrosol and alcohol as an indirect promoter of endogenous hydroxytyrosol generation. This trial was registered at controlled-trials.com/isrctn/ as ISRCTN 35739639.