Preclinical safety evaluation of Zizyphi spinosi semen simple crushed powder as a potential anti-dementia ingredient

Zizyphi spinosi semen (ZSS), the seeds of Zizyphus jujuba Miller var. spinosa, is a well-known ingredient in Chinese herbal medicine that has been used for centuries to treat insomnia and anxiety. ZSS is usually served as a decoction in traditional Chinese medicine; however, our previous study using dementia model mice revealed that the anti-dementia activity of ZSS is enhanced by simple crushing rather than hot water extraction, with an effective dose of 3.3 mg/kg. To assess the safety of ZSS simple crushed powder (ZSS–SCP, Ziziphas PowderTM), we conducted a series of preclinical safety assessments, including in vitro mutagenicity studies (Ames tests) in bacteria, and acute and 90-day sub-chronic oral toxicity studies in rats. In the acute toxicity study, ZSS–SCP was administered at a dose of 2,000 mg/kg to five male and five female animals. In the sub-chronic toxicity study, ZSS–SCP was tested at doses of 300 and 1,000 mg/kg, with 10 male and 10 female animals for each dose. The results showed no ZSS–SCP-induced mutations, animal deaths, or clinical abnormalities, indicating a No Observed Adverse Effect Level of 1,000 mg/kg/day. Our findings suggest that ZSS–SCP is a safe ingredient in anti-dementia foods.