¹Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Eastern Mediterranean University, Famagusta, TRNC, via Mersin 10 Turkey DOI : 10.29228/jrp.480 Urolithins are the bioavailable metabolites of ellagitannins, abundantly available in dietaries routinely consumed. The number of research studies growingly enlarges on the beneficial effects of these natural chemicals on human health. From this perspective, one aspect of scientific interest in urolithins covers the development of techniques for their qualitative and quantitative analysis in diverse samples. UV-based and tandem-mass spectroscopic analyses have been the major ones used and applied so far. However, the fluorescent characteristics of these molecules have not received sufficient attention. Within the scope of this research study, major urolithins (Urolithin A and B) concomitant to their methyl ether metabolites have been synthesized and their quantitative analysis has been investigated via an HPLC system connected to fluorescence detection. The results displayed the superior and comparable characteristics of the methodology developed against the previous techniques. Keywords : Urolithin A; Urolithin B; Urolithin methyl ethers; Fluorescence; HPLC; quantification