Objective The aim of the present study was to investigate hyaluronic acid (HA) concentrations in vocal folds among patients with Reinke’s edema. Study Design Prospective and experimental study. Setting Single tertiary center. Methods An HA binding protein isolated from bovine nasal cartilage was used to identify and isolate the HA from samples. Plates coated with biotin-conjugated binding protein and streptavidin-europium conjugate were sequentially incubated with 18 Reinke’s edema samples and 11 female vocal fold cover samples from cadavers (the superficial layer of the lamina propria; control group). After the release of europium from streptavidin in enhancement solution, final fluorescence was measured in a fluorometer. Results The mean HA concentration in Reinke’s edema vocal folds was significantly higher than that in the control vocal folds (9.2 × 103 vs 0.9 × 103µg/g). Conclusion Vocal fold covers affected by Reinke’s edema present a higher concentration of HA than do vocal fold covers with no edema.