Ultraviolet-B (UV-B) radiation is a key but under-researched environmental factor that initiates diverse responses in plants, potentially affecting their distribution. To date, only a few macroecological studies have examined adaptations of plant species to different levels of UV-B. Here, we combined herbarium specimens of Hieracium pilosella L. and Echium vulgare L. with a novel UV-B dataset to examine differences in leaf hair traits between the plants’ native and alien ranges. We analysed scans of 336 herbarium specimens using standardized measurements of leaf area, hair density (both species) and hair length (H. pilosella only). While accounting for other bioclimatic variables (i.e. temperature, precipitation) and effects of herbivory, we examined whether UV-B exposure explains the variability and geographical distribution of these traits in the native (Northern Hemisphere) vs. the alien (Southern Hemisphere) range. UV-B explained the largest proportion of the variability and geographical distribution of hair length in H. pilosella (relative influence 67.1%), and hair density in E. vulgare (66.2%). Corresponding with higher UV-B, foliar hairs were 25% longer for H. pilosella and 25% denser for E. vulgare in records from the Southern as compared to those from the Northern Hemisphere. However, focusing on each hemisphere separately or controlling for its effect in a regression analysis, we found no apparent influence of UV-B radiation on hair traits. Thus, our findings did not confirm previous experimental studies which suggested that foliar hairs may respond to higher UV-B intensities, presumably offering protection against detrimental levels of radiation. We cannot rule out UV-B radiation as a possible driver because UV-B radiation was the only considered variable that differed substantially between the hemispheres, while bioclimatic conditions (e.g. temperature, precipitation) and other considered variables (herbivory damage, collection date) were at similar levels. However, given that either non-significant or inconclusive relationships were detected within hemispheres, alternative explanations of the differences in foliar hairs are more likely, including the effects of environment, genotypes or herbivory.