AIMS: In this study, the effects of licorice flavonoid oil (LFO) supplementation on mobility functions were evaluated in middle-aged and older women who underwent daily physical exercise. METHODS: The 73 women aged 59-85 years (71.2 ± 5.2 years) were randomly assigned to the LFO group (n = 37) or the placebo group (n = 36). For 16 weeks, the LFO group consumed a daily capsule containing 300 mg of LFO, while the placebo group consumed a placebo capsule. All participants were instructed to complete a strength training program during the 16 weeks and to increase their daily step count by 1000. 10-m walking speed (with/no obstruction), one-leg standing time with eyes open, handgrip strength, isometric knee extension strength, and body composition were evaluated at baseline and every eight weeks. RESULTS: In the 10-m walking speeds (with/no obstruction), LFO supplementation did not show significant improvements. One-leg standing time was significantly prolonged with LFO intake (LFO: baseline 73.9 s vs 16 weeks 93.5, placebo: baseline 82.8 vs 16 weeks 87.1, p = 0.03). In addition, a significant decrease in BMI and body fat percentage with LFO was found (p = 0.01, p = 0.03, respectively). DISCUSSION: Since a lower BMI corresponds to a lighter physical load on the lower limb, in addition, since LFO might improve skeletal muscle function by antioxidant activity, participants could stand longer and body balance control was improved. CONCLUSION: LFO supplementation improved body balance control and may contribute to fall prevention in healthy middle-aged and older women having daily physical exercise. TRIAL REGISTRATION: UMIN Clinical Trial Registry No. 000029712.