IntroductionHealth literacy is an important predictor of health behavior and self-rated health, playing a crucial role in shaping public health outcomes. Valid and reliable health literacy assessments are essential for effectively tailoring health interventions, particularly in different cultural contexts. Several questionnaires have been developed to measure health literacy, including the widely used 47-item Health Literacy Questionnaire and its shorter versions. However, the validity of these shorter and more time efficient HLS-EU versions, such as the 16-item HLS-EU-Q16, has been less extensively researched in different age subgroups. Given Slovenia’s aging population and the differences in health literacy between adults and older adults, it is important to assess whether the HLS-EU-Q16 performs reliably across these groups. Therefore, the present study aimed to examine the validity and reliability of the 16-item health literacy questionnaire (HLS-EU-Q16) in Slovenia across two age groups: adults and older adults.MethodsWe analyzed representative Slovenian sample data from the Health Literacy of Adults study. The sample included 2,327 adults aged 23–64 years (53.7% women) and 876 older adults aged 65–99 years (55.7% women).ResultsConstruct validity revealed a modified three-factor structure of the HLS-EU-Q16 among adults, reflecting a broader conceptualization of health literacy that encompasses overlapping domains of health promotion, disease prevention, and health care. In contrast, a two-factor structure emerged among older adults, indicating a more consolidated approach where these domains are perceived as part of an integrated health management strategy. This suggests that adults may conceptualize health literacy across multiple, yet interconnected dimensions, while older adults approach it more holistically. Convergent validity, reliability, concurrent-predictive, concurrent, and discriminant validity results were satisfactory for both age groups. Predictive validity, however, provided somewhat inadequate results, as the tool poorly predicted certain health behaviors, such as smoking, alcohol consumption, and physical activity, in both age groups.ConclusionOverall, the HLS-EU-Q16 demonstrated adequate validity and reliability among both adults and older adults, indicating that it is an appropriate instrument for assessing health literacy in Slovenia.