| Objective:The natural hirudin?100AT-U/g?extracted from the living body of Hirudinaria manillensis was used as the research object to test its antioxidant activity.Natural hirudin emulsion and eye cream were developed based on formula design theory,and its anti-aging and improving eye microcirculation effects and mechanism were discussed,providing ideas and experimental reference for its application in the field of cosmetics.Method:1.The ability of natural hirudin to remove DPPH·,·OH,·O2-was determined by Vit C positive control method,and analyze its antioxidant activity.2.Emulsions and eye creams were evaluated by sensory evaluation,high and low temperature cycling test,centrifugation test,p H value and viscosity test,and microbial challenge test.The formulation relationship of each system and the amount of natural hirudin added were analyzed.3.The multi-factor orthogonal test and single-factor test results of emulsion were evaluated by fuzzy comprehensive evaluation method,and the effect of process conditions on the emulsification effect was analyzed.4.Objective instrument was used to evaluate the efficacy of natural hirudin in moisturizing and anti-wrinkle.The experimental group was composed of 3%natural hirudin emulsion and the blank group was composed of matrix emulsion.The changes of MMV value and TEWL value were measured before and after using the emulsion for 4h and 28d.5.Subjects were randomly divided into two groups by self-evaluation:the experimental group and the control group,each with 20 subjects.During the experiment,natural hirudin eye cream containing 3%and Vit C eye cream containing 3%were used respectively.The experiment period was 60 days,and the subjects were asked to self-evaluate the four items of eye fine lines,skin smoothness,dark circles and edema by means of questionnaire survey,and count the effective rate.The practical application effect of natural hirudin and Vit C was compared and analyzed.6.Using human trials assessment method,the right eye of one subject was selected as the experimental group and the left eye as the control group.During the experiment,natural hirudin eye cream containing 3%and similar eye cream of a well-known brand were used.Pouch size was measured at 0,10,20,30,40,50,60 days,and the depth of dark circles and the reduction of fine lines were compared.At the same time,periocular photographs before and after use were collected.To compare and evaluate the anti-aging and microcirculation effects of natural hirudin,and analyze its mechanism of action.Result:1.The IC50 values of natural hirudin for DPPH·,·OH and·O2-removal were 14.1mg/m L,4.2 mg/m L and 21.1 mg/m L,respectively(IC50 values of control Vit C were 2.6mg/m L,0.43 mg/m L and 0.032 mg/m L,respectively).Its antioxidant activity was generally lower than that of control Vit C,and its ability to scavenge DPPH·and O2-was lower than that of Vit C,but its ability to scavenge·OH was strong,only 1/10 times that of extremely strong Vit C.2.Dosage ratio relationship of each emulsion system:soybean lecithin?1.0%?and Seppic-Montanov L?1.5%?were selected as emulsifying system;sclerotium gum?0.1%?and Carbopol 940-arginine?0.2%?were selected as thickening system;potassium sorbate?0.2%?and 1,2 hexanediol?0.5%?were selected as antiseptic system;the addition amount of natural hirudin should not exceed 4.0%.Carbopol 940?0.30%?and arginine?0.30%?were selected as the thickening system of eye cream;P-Hydroxyacetophenone?0.50%?and hexanediol?0.50%?were used as the antiseptic system;and natural hirudin was added in 3.0%.The eye cream passed the anticorrosion challenge test with good stability and good sensory evaluation?double blind?of 8 indicators.3.The results of orthogonal experiment show that the factors affecting the emulsion formulation system are in order:homogenization time>homogenization speed>emulsification temperature.In combination with the single-factor experiment of homogenization time,the optimal process conditions of the emulsion were determined:the emulsification temperature was 80?,the homogenization time was 5min,and the homogenization speed was 4000r/min.4.Results of the blank control group:the MMV growth rate and TEWL loss rate of the experimental group and the blank group were basically the same within 4 hours,and there was no significant difference in the moisture retention performance between the two groups in a short time.The MMV growth rate of the experimental group was 36.59%before and after 28d,which was significantly higher than that of the control group.The loss rate of TEWL was-21.69%,which was significantly lower than that of the control group,indicating that natural hirudin has a long-term moisturizing performance and an antioxidant effect.5.Results of Vit C control group:The effective rates of four items of reducing fine lines of eyes,smoothing sensation of skin,improving dark circles and improving edema effect were 65.0%,80.0%,45.0%and 40.0%in the test group,and 35.0%,60.0%,15.0%and 5.0%in the Vit C control group,respectively.The anti-aging and improving microcirculation effects of the test group were significantly better than those of the control group,which also indicated that natural hirudin antioxidant The effect of in vivo application is more prominent than that of Vit C.6.Results of the market control group:the effect of improving edema and reducing fine lines in the experimental group was significantly better than that in the control group.Both had little difference in improving dark circles.It is proved that natural hirudin has the effect of improving eye microcirculation and anti-aging,and has certain advantages compared with similar products.Conclusion:1.The antioxidant activity of natural hirudin on a single reaction system was worse than that of Vit C.2.Natural hirudin has good compatibility and stability in cosmetic system,but the addition amount should not exceed 4.0%.3.At the same amount of addition,the in human application effect of natural hirudin antioxidant efficacy was better than that of Vit C,and there was a difference between the results of chemical method and in human application effect.4.The results of objective instrument evaluation,subject self-evaluation and human trial evaluation were consistent,indicating that natural hirudin has the functions of moisturizing and anti-wrinkle,antioxidation,anti-aging and improving eye microcirculation. |
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