Abstract: Objective To establish abnormal toxicity test and pyrogen test of glycosaminoglycan from sea cucumber for injection. Methods The median ethal dose (LD₅₀) and 95% confidence limit of glycosaminoglycan from sea cucumber for injection were obtained by referring to guiding Principles for Application of Injection Safety Test method in Pharmacopoeia of the people’s Republic of China, Volume IV, 2020 edition, and the limit of abnormal toxicity test was determined. A methodological study on pyrogen test of glycosaminoglycan from sea cucumber for injection was studied. Results The LD₅₀ of this product was 189.4 mg·kg^-1 for intravenous injection, and the 95% confidence limit was 239.0~139.8 mg·kg^-1. 1/4 of the lower confidence limit (139.8 mg·kg^-1) was taken as the limit of abnormal toxicity, that was, 35 mg·kg^-1 was injected into the tail vein of mice. The product was prepared with sodium chloride injection into a solution with a concentration of 1.4 mg·mL^-1, and examined according to the abnormal toxicity test method 1141, Pharmacopoeia of the People’s Republic of China, Volume IV, 2020 Edition. As the results, all the mice were in normal state after injection, and no death was observed in 48 hours. Glycosaminoglycan from sea cucumber for injection was prepared with sodium chloride injection in a solution of 1 mg·mL^-1 at a dose of 2 mg·kg^-1 of rabbits body weight (about 5 times of the clinical dosage), which had no interference on the increase of rabbits’ body temperature caused by bacterial endotoxin. Conclusion 35 mg·kg^-1 dose can be used as the limit of abnormal toxicity; the pyrogen of this product can be controlled by rabbits method, and 2 mg·kg^-1 dose is used as the limit for pyrogen examination.

Key words: glycosaminoglycan from sea cucumber, injection, mice, abnormal toxicity, rabbits, pyrogen