Hesperidin is a flavonoid with many nutritional benefits including antioxidant activity in food formulations; however, hesperidin is practically insoluble in water. A commercial enzymatic process has been developed in which a glucose molecule is attached to hesperidin increasing the solubility by approximately 100,000 times. The substance is called glucosyl hesperidin (GH) with the main component being monoglucosyl hesperidin (MGH; 75 to 85%). This paper presents results of OECD-compliant toxicity studies with GH, including 4-week and 13-week sub-chronic toxicity, and teratogenicity studies in rats, and chromosomal aberration and mouse micronucleus formation tests. There were no deaths and no treatment-related adverse effects in the 4-week (highest dose 15,000 ppm) or the 13-week sub-chronic (highest dose 50,000 ppm) studies. There were no statistically significant treatment-related adverse effects on any parameter evaluated. The NOEL in the 4-week study was calculated as 1,280 mg/kg/day in females and 1,206 mg/kg/day for males, and in the 13-week study, the NOEL was 3,428 and 3,084 mg/kg/day, for females and males, respectively. In the teratogenicity study, the NOAEL was 1,000 mg/kg/day of treatment for both dams and fetuses. No genotoxicity was observed in the chromosomal study at 5,000 μg/mL and no micronuclei at 2,000 mg/kg, respectively. The results of these OECD-compliant studies support the safe use of GH as a food and beverage ingredient.