Study On The Application Of Active Peptides Of Andrias Davidianus In Cosmetics

Andrias davidianus,commonly known as"giant salamander",is a large amphibian unique to China,distributed in Hunan,Hubei,Shaanxi and other provinces and regions,containing a variety of bioactive substances,and has broad prospects for development in food,health food and daily chemical products.At present,the breeding technology of Andrias davidianus is fully mature,and the number of cultured Andrias davidianus has increased rapidly.However,the application of Andrias davidianus is still in the fresh food stage,and there is little research on the intensive processing and utilization of Andrias davidianus in the daily chemical industry.In this paper,the preparation,purification and peptide synthesis of oligopeptides from Andrias davidianus were carried out,and their structural characteristics,biological activity comparison and application in facial mask were studied.The main contents are as follows:（1）Study on bioactivity of oligopeptides of Giant Salamander.The oligopeptides of Giant Salamander were prepared by enzymatic hydrolysis.After separation and purification of glucan LH-20,two components of ADOP1 and ADOP2 were obtained.The results show that:Oligopeptides could clear DPPH·,ABTS⁺·,OH·and O₂^-·,and the DPPH·clearance rates were 83.83±0.45%,ABTS⁺·clearance rates were 71.91±0.34%,OH·clearance rates were 38.34±1.70%and O₂^-·clearance rates were 86.65±0.59%.The scavenging ability was O₂^-·>DPPH·>ABTS⁺·>OH·;The antioxidation ability of oligopeptides of Salamander was compared with different kinds of bioactive peptides,different brands of peptides and different isolated and purified components,indicating that the oligopeptides of Salamander had strong antioxidant ability.The oligopeptides of giant Salamander had inhibitory effect on tyrosinase.The tyrosinase inhibitory ability was compared with that of different isolated and purified components,which proved that it had strong inhibitory effect on tyrosinase.The oligopeptides of salamander showed no inhibitory effect on Proteus and other bacteria.RAW264.7 Mouse macrophages were co-cultured with giant Salamander oligopeptides,and it was found that giant Salamander oligopeptides had proliferative effects on the growth of RAW264.7mouse macrophages.（2）Study on the structure characterization of oligopeptides of Giant Salamander and the comparative verification of the bioactivity of synthetic peptides.By Fourier infrared spectroscopy analysis of oligopeptides of Giant Salamander,the existence of amide functional groups and imidazole functional groups,containing hydroxyl,ether,β-folded structure and random curling secondary structure was identified.It was identified as oligopeptide by mass spectrometry.A total of 81 peptides were identified by sequence analysis,and the top three peptides were screened for peptide synthesis.Three peptides,SR-16,IR-7 and VK-14,were obtained by solid-phase synthesis,HPLC chromatography and mass spectrometry,and their biological activities were verified.It was found that the antioxidant capacity of the three peptides was SR-16>VK-14>IR-7.The inhibitory ability of tyrosinase SR-16>IR-7>VK-14 and the proliferative effect of tyrosinase on RAW264.7mouse macrophages were VK-14>SR-16>IR-7,and the oligopeptides were not as strong as those of Giant Salamander.The results showed that the oligopeptide of SR-16 and Giant Salamander had no inhibition zone against Proteus,Bacillus subtilis,Staphylococcus aureus,Escherichia coli,Enterobacter aerogenes and Shiwanella.IR-7 solution of 200mg/m L had obvious inhibition zone against the above six bacteria,and the inhibition zone diameter decreased gradually with the decrease of the concentration.VK-14 solution of200mg/m L had a bacteriostatic zone on Bacillus subtilis and Shiwanella,but the bacteriostatic effect was not as obvious as that of IR-7.（3）The whitening mask was prepared with oligopeptide of Giant Salamander as raw material.The optimal formula of the mask was determined by single factor test with moisture rate,drying time and viscosity as indexes.The physical and chemical indexes,stability and sensory evaluation of the mask were studied.The results showed that glycerol 5%,polyvinyl alcohol 0.5%,sodium carboxymethyl cellulose 1%,butanediol 4%,sodium hyaluronate 0.5%,triethanolamine 0.5%,carbomer 0.5%,allantoin 0.5%,giant salamander oligopeptide 0.5%,methyl paraben and flavor appropriate amount,deionized water added to 100%is the best formula of whitening mask;The drying time of the mask is 960s,the viscosity is between 74～76 Pa·s,and the p H range is 5.8～6.3,all of which are in line with national standards.The mask has good heat resistance,cold resistance and hot and cold alternations,no stratification phenomenon after centrifugation,good stability;Before use,the mask is clear and transparent,good color,no obvious odor,fluidity and not greasy when used,after use,the skin is white and tender as cream,and the effect is mild.