Preparation Of Shuangyang Sika Deer Fetal Peptide And Application Of Anti-skin Damage Mask

Compendium of Materia Medica recorded that deer fetus could be used to treat women’s strain and have black and rough facial skin.In our study,we aimed to develop deer fetus’industrial application.With the placenta of shuangyang sika deer as raw material,we used its peptide product to make facial mask.The research results were showed as following:The optimal extraction process of Shuangyang sika deer fetus protein was determined by single factor orthogonal design.The enzymatic hydrolysable processing condition of deer fetus protein was optimized by response surface method.The molecular weight and amino acid composition were determined by SDS-PAGE electrophoresis and amino acid analyzer.Meanwhile,deer fetus peptides were separated and purified by ultrafiltration and gel filtration chromatography.The results showed that:The optimal ultrasonic extraction process of deer fetus protein was 30℃,300 W,40 min and material-liquid ratio at 1:40.After screening,compound protease was identified as a tool enzyme.The composite protease is composed of papain,neutral protease and alkaline protease according to the enzyme activity ratio of 1:1:1.The optimum hydrolyzed process of deer fetoprotein was51℃,p H 7.0,4900 U/g enzyme and 3 h zymolytic time.The molecular weight of the hydrolysates was mainly distributed in 10 k Da～17 k Da.Deer fetus hydrolyzed products are composed of 18 amino acids,of which antioxidant amino acids and hydrophobic amino acids each account for 14.40%and 39.50%,respectively.The proteolysis product of deer embryo had two characteristic absorption peak at 220 nm and 280 nm.Three groups of polypeptide components with molecular weight between 5.8～10 k Da were obtained by separation and purification,which were named as DFP1、DFP2、DFP3.Their molecular weight order is DFP1>DFP2>DFP3.The anti-skin damage activity of Shuangyang sika deer fetus polypeptide was investigated by in vitro antioxidant test,tyrosinase inhibition experiment,moisturizing experiment,NIH3T3 cell proliferation and anti-inflammatory experiment.The results showed that:Deer fetus peptides had anti-oxidative damage activity,DFP1、DFP2 and DFP3 had scavenging ability against·OH free radicals,O₂^-free radicals and ABTS⁺free radicals,of which DFP3 had stronger ability.The IC₅₀of DFP3 on 3 free radicals were0.703 mg/m L,1.256 mg/m L and 1.101 mg/m L,respectively,but DFP2 has the weakest oxidation resistance.Deer fetus polypeptide had anti-light damage activity.DFP1,DFP2and DFP3 had certain inhibitory capacity on tyrosinase.Moreover,DFP3 had the strongest inhibition ability and the maximum inhibition rate reached 61.13±1.83%.Deer fetus peptide had anti-drying effect.DFP3 has higher hygroscopicity and moisture retention than DFP1 and DFP2.At 48h,the moisture absorption rate of DFP3 were:14.46±1.02%（RH=32%）and 29.96±1.80%（RH=81%）,the moisture retention rate of DFP3 were35.01±0.50%.Deer fetus peptides can promote skin healing.DFP1,DFP2 and DFP3 all promoted the proliferation of NIH3T3 cells and inhibited the secretion of NO,among which DFP3 is the best one.The cell proliferation rate and NO secretion of DFP3 were:155.42±1.77%and 80.29±0.87 pg/m L,which showed that deer fetus peptides had no obvious toxicity and had certain anti-inflammatory and skin healing effects.The anti-skin injury mask was prepared from DFP3.The toxicity and irritation of the mask were investigated by acute toxicity test on mice and acute percutaneous test on mice.Meanwhile,the physical and chemical indexes,stability and sensory evaluation of the mask were studied.The results showed that:The best formula for the mask is the addition of 12%DFP3,5%glycerin,0.5%triethanolamine,3%polydimethylsiloxane,3%PEG-40 stearate,0.3%hyaluronic acid Sodium and 0.6%carbomer.Appropriate amount of p-hydroxyacetophenone and essence,deionized water was added to 100%.The mask never caused death,poisoning or skin irritation in mice.Indicating that it was non-toxic and irritating to mice.The p H range of the mask was 6.0～6.5,the viscosity was between75～76 Pa·s and the drying time was 16 minutes,which all followed the relevant regulations.The mask had good heat resistance and cold resistance.At the same time,there was no delamination after centrifugation.the mask had good appearance and light and the skin felt smooth after using the mask.