The skin of puffer fish is rich in collagen,which can be a high-quality preparation material for collagen peptide.The deep processing of fish skin can not only reduce the waste of resources but also expand the processing mode of puffer fish in China.In this research,Fugu bimaculatus collagen peptide (FBCP) was prepared by conducting a single factor experiment and an orthogonal experiment,in which,the cosmetic efficacy and skin irritation of FBCP were studied.The optimum enzymatic hydrolysis conditions of FBCP by alkaline proteinase were determined as follows:the solid-liquid ratio was 1∶10,with the enzymolysis temperature as 50 ℃,8 000 U/g of quantity of enzyme,with enzymolysis pH as 9 and the enzymolysis time as 4 h.The peptide yield of FBCP from the skin is 39.65% under these conditions.Ultrafiltration membranes were selected and four components of FBCP with different molecular weights,namely,FBCP1 (Mw<1 kDa),FBCP2 (1 kDa<Mw<5 kDa),FBCP3 (5 kDa<Mw<10 kDa),and FBCP4 (Mw>10 kDa),were separated,respectively.Besides,the result showed that the hygroscopicity and moisture retention of FBCP1 were better than those in other three components.Moreover,the results of antioxidant test suggested that the FBCP1 radical scavenging rate was the highest,with the IC₅₀ of O₂^－·,·OH and DPPH radical scavenging tests as 1.659,5.582 and 1.801 mg/mL,respectively.Multiple skin irritation and acute eye irritation experiments showed that application of up to 50% (W/V) FBCP was not irritating to the rabbit skin and rabbit eyes.In summary,FBCP has excellent anti-oxidation and moisturizing effects,along with non-irritating to skin and eyes,thus capable of providing a basic reference for its application in cosmetics.