This study evaluated the sub-chronic oral toxicity of freeze-dried deer velvet antler (DVA) extract in ICR mice over a 90-day period. The extract was administered orally to male and female mice at doses of 1,000, 2,000, and 5,000 mg/kg body weight daily; the control group (0 mg/kg body weight) received vehicle only. No clinical signs of toxicity or mortality were observed. Body weight gain, food consumption, and general behavior remained normal across all groups. Hematological analysis revealed no dose-dependent alterations in hemoglobin levels, platelet characteristics, or red or white blood cell counts. Males administered 2,000 mg/kg DVA exhibited a slight increase in reticulocyte count, whereas females exhibited reduced count at 5,000 mg/kg; however, neither finding was doseresponsive or toxicologically significant. Serum biochemistry including liver and kidney function markers (ALT, AST, -GT, BUN, and creatinine) remained within normal range, and no evidence of organ damage was observed. Notably, male mice exhibited a significant reduction in serum triglycerides compared with those in the control group, suggesting potential lipid-modulating effects of DVA; however, this finding requires further investigation. Based on these findings, the no-observed-adverse-effect level (NOAEL) of DVA extract was determined to be greater than 5,000 mg/kg/day in both sexes. These results support the safety profile of DVA extract at the tested doses, and its potential metabolic benefits.

Velvet antler is widely used as a traditional medicine, and numerous studies have demonstrated its tremendous nutritional and medicinal values including immunity-enhancing effects. This study aimed to investigate different deer velvet extracts (Sample 1: raw extract, Sample 2: dried extract, and Sample 3: freeze-dried extract) for proximate composition, uronic acid, sulfated glycosaminoglycan, sialic acid, collagen levels, and chemical components using ultra-performance liquid chromatography-quadrupole-time-of-light mass spectrometry. In addition, we evaluated the cytotoxic effect of the deer velvet extracts on BV2 microglia, HT22 hippocampal cells, HaCaT keratinocytes, and RAW264.7 macrophages using the cell viability MTT assay. Furthermore, we evaluated acute toxicity of the deer velvet extracts at different doses (0, 500, 1000, and 2000 mg/kg) administered orally to both male and female ICR mice for 14 d (five mice per group). After treatment, we evaluated general toxicity, survival rate, body weight changes, mortality, clinical signs, and necropsy findings in the experimental mice based on OECD guidelines. The results suggested that in vitro treatment with the evaluated extracts had no cytotoxic effect in HaCaT keratinocytes cells, whereas Sample-2 had a cytotoxic effect at 500 and 1000 μg/mL on HT22 hippocampal cells and RAW264.7 macrophages. Sample 3 was also cytotoxic at concentrations of 500 and 1000 μg/mL to RAW264.7 and BV2 microglial cells. However, the mice treated in vivo with the velvet extracts at doses of 500–2000 mg/kg BW showed no clinical signs, mortality, or necropsy findings, indicating that the LD50 is higher than this dosage. These findings indicate that there were no toxicological abnormalities connected with the deer velvet extract treatment in mice. However, further human and animal studies are needed before sufficient safety information is available to justify its use in humans.