Preparation And Antioxidant Activity Of Collagen Peptides From Antler Bone Tissue And Their Application In Cosmetics

Oxidative stress poses a serious threat to human health in modern society.Bioactive peptides with antioxidant activity,derived from biological raw materials like collagen,are gradually attracting attention due to their high safety,efficacy,and ease of absorption and accessibility.Antler is the only fully regenerable organ in mammals,and it grows very rapidly,up to 2cm/day.However,its roots are heavily ossified,making it difficult to make full use of the traditional hot-fried processing method.Studies have indicated that the collagen content of antler bone tissue is very rich,and that there is little risk of contamination,with few restrictions on the use of deer-derived products.Thus,deer antler bone tissue is a promising source of bioactive peptides.Studies have shown that deer antler bone tissue contains a lot of collagen and that products derived from deer have low contamination risk and few usage restrictions.Therefore,deer antler bone tissue is a good source of bioactive peptides.In this study,collagen was extracted from deer antler ossified tissues using acetic acid and pepsin,respectively.The collagen obtained by the two methods was then hydrolyzed using neutrase,alcalase,trypsin,flavoenzyme,and papain.The collagen peptides with the highest antioxidant activity were purified using ultrafiltration and gel chromatography,and the amino acid composition of the peptides was examined.The results showed that neutrase was the most effective hydrolytic enzyme and that pepsin was used to extract collagen.The optimum enzyme digestion process was then optimized to obtain the enzyme dosage of 3200 U/g,the enzyme digestion temperature of 52℃,the p H of 6.1,the substrate concentration of 10%,and the enzyme digestion time of 3 hours.Under these conditions,the DPPH clearance rate of the enzyme digestion product was 73.41±0.48%.The strongest antioxidant activity of the collagen peptide was obtained after purification,with the DPPH clearance rate of90.58±1.27%.Analysis of the amino acid composition showed that this collagen peptide had a high proportion of amino acids with antioxidant capacity.To determine the antioxidant activity of the collagen peptides obtained from antler ossified tissues,a comprehensive assay was conducted at both the in vitro and cellular levels.The in vitro assay focused on the ability of the collagen peptide to scavenge common free radicals and inhibit oxidative damage to plasmid DNA,while the cellular assay examined the effect of the collagen peptide on the oxidative damage model of HaCaT cells,which were constructed using H₂O₂.The in vitro results revealed that the collagen peptides exhibited the DPPH clearance rate of 90.66%,the superoxide anion radical clearance rate of 78.33%,the ABTS clearance rate of99.59%,and the hydroxyl radical clearance rate of 83.34%.Additionally,the peptides inhibited oxidative damage to plasmid DNA.At the cellular level,the collagen peptides exhibited the ability to protect HaCaT cells from oxidative damage,maintain cell viability,and significantly reduce the accumulation of intracellular ROS and MDA.Moreover,the activity of antioxidant enzymes such as CAT,SOD,and GSH-Px increased.The three cosmetic products were designed using antler collagen peptides as the core material.These products underwent various tests such as sensory testing,stability testing,microbiological testing,physical and chemical testing,and toxicological testing.The results confirmed that the three cosmetic products had good skin feel,stability,and hygiene.They also met the requirements for heavy metal and harmful substance content as per the Technical Specification for Cosmetic Safety.Furthermore,they were not found to be irritating to the skin.Consequently,the three domestic non-special use cosmetic products successfully obtained registration numbers.In summary,this study established the enzymatic digestion process for collagen peptides derived from antler bone tissue with strong antioxidant activity,as well as the separation and purification process based on ultrafiltration and gel chromatography.The study investigated the antioxidant activity of collagen peptides from antler at both in vitro and cellular levels.Furthermore,three cosmetic products containing collagen peptides from antler were designed,tested for safety,and received cosmetic registration numbers.These findings provide a new idea for the development and application of antler products and bioactive peptides with antioxidant capacity.