| Puffer fish resources are rich in our country.The skin of puffer fish is rich in collagen,while collagen hydrolysate collagen peptide has the activity of antioxidation,inhibiting the activity of angiotensin converting enzyme,inhibiting the activity of tyrosinase,skin care and anti-tumor.In this case,it is a good source of green raw materials for medical and health care products.However,since now,there are few research carried out on cosmetic and skin care.It is urgent to develop the available value of fish skin collagen peptide in skin care.In this paper,the extraction process of collagen peptide from the skin of Fugu bimacuJatus was studied.Then the collagen peptide with different molecular weights was prepared by ultrafiltration separation technology,and the effectiveness and safety of which were studied at last.The work were divided into three parts.1.Extraction of the collagen peptide from the skin of the Fugu bimacuJatus.The effects of5 single factors,such as enzymatic hydrolysis temperature,enzyme hydrolysis time,enzyme hydrolysis pH,enzyme addition and solid-liquid ratio,were investigated by alkaline protease hydrolysis of fish skin of Fugu bimacuJatus.Through the orthogonal experiment,the optimum enzymatic hydrolysis conditions of collagen peptide were determined as follows: the solid-liquid ratio was 1:10;the enzymolysis temperature was 50;the enzyme was added 8000 U/g;the enzymolysis was pH 9 and the enzymolysis time 4 h.Under these conditions,the peptide yield of collagen protein from the skin is 39.65%.2.Study on the effectiveness of collagen peptides from the skin of Fugu bimacuJatus.10 kDa,5 kDa,and 1 kDa ultrafiltration membranes were selected,and four components of fish skin collagen peptide with different molecular weights FBCP1(Mw<1 kDa),FBCP2(1kDa<Mw<5 kDa),FBCP3(5 kDa<Mw<10 kDa),and FBCP4(Mw>10 kDa),were separated,respectively.The antioxidant activity,hygroscopicity,moisture retention of four kinds of fish skin collagen peptides and their effects on preventing L929 cells from radiation damage by UVB were investigated.The results of vitro antioxidant test showed that the FBCP1 radical scavenging rate was the highest.The clearance rates were 72.85%,90.22% and 82.10% in the O2-·,·OH and ·DPPH radical scavenging tests,respectively,and 1.659 mg/mL,5.582 mg /m Land 1.801 mg/mL respectively.In the test of moisture absorption and moisture retention,the moisture absorption rate and moisture retention rate of FBCP1 in RH=70% and RH=40%environment are better than the other three components.The moisture absorption rate and moisture retention rate in the RH=70% environment are 4.539% and 0.932% respectively.The moisture absorption rate and moisture retention rate in RH=40% environment are 4.125% and0.897% respectively,and in the skin water diversion loss test,the TWEL of the 20%concentration of FBCP1 components is 8.9,the moisturizing effect of which is obviously higher than the other three components and tends to positive control level.In the prevention of L929 cell UVB radiation damage test,FBCP1 has the ability to prevent the light damage of L929 cells,which provides a solid theoretical and experimental basis for the development of follow-up cosmetics.Compared with the UVB group,the damage degree of L929 cells was obviously reduced by adding FBCP1 before irradiation,and the cell vitality increased by 60.09% when the concentration was 40 mg/mL,which proved that FBCP1 had the ability to prevent the light damage of L929 cells.Compared with the UVB group,after adding FBCP1,the cell viability of each concentration group of FBCP1 decreased,and the activity of 40 mg/mL decreased by14.93% at the concentration of 40 mg/mL,which showed that FBCP1 did not have the ability to repair the light damage of L929 cells.3.Evaluation of the irritation of collagen peptides from the skin of the Fugu bimacuJatus.The average relative survival rate of EpiSkin skin model stimulation test was 93% after smearing1.5g/mL FBCP1 solution.After adding the 1.5g/mL FBCP1 solution into the opacification of the bovine cornea,the score of the bovine corneal reaction was-0.187±0.044?3.Both of the two experimental results showed that the FBCP1 was not irritating and feasible of being a cosmetic material... |
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