Research On Screening, Extraction And Mechanism Of Active Materials In Cosmetic Anti-allergy And Anti-irritation

Skin allergy and irritation were an immune response appeared to physiological disorder or abnormal tissue trauma when the body was stimulated by antigens, consisted of allergic contact dermatitis (ACD) and irritant contact dermatitis (ICD). According to the mechanisms of ACD and ICD, 25 plant extracts with anti-allergy and anti-irritation effects were screened. The main functional ingredient content, extraction and process optimization, function pathway and mechanism were also studied.The irritation, anti-allergy and anti-irritation effects of 25 plant extracts were determined respectively by red blood cell (RBC) hemolysis test, in vitro hyaluronidase inhibition and improved RBC hemolysis test. Opuntia Milpa Alta powder extract was proved to have the best anti-allergy and anti-irritation effects.The results of Opuntia Milpa Alta on RBC hemolysis rate, in vitro hyaluronidase inhibition rate and preventive effects of SDS- induced RBC damage were 3.1%, 97.1%, 46.4%. Chemical composition analysis revealed that Opuntia Milpa Alta polysaccharides, about 73.9% of soluble solids content, were the major components.Opuntia Milpa Alta polysaccharides contents, soluble solids content and conductivity were compared between the methods of aqueous extraction and enzyme-assisted aqueous extraction. The results indicated that cellulase hydrolysis can increase the dissolution rate of polysaccharide about 15% and protease can increase the total soluble solids content about 18.5%. However, enzyme-assisted aqueous extraction resulted in ion intensity increased significantly, compared with aqueous extraction by 23%. In the case of single factor, water to solid ratio 15²5, temperature 40⁶0℃, time >1h, the higher polysaccharides rate was obtained. Based on the single factor experiments results and practical operation, the extraction procedure was optimized using Box-Behnken design and Surface Response Methodology (RSM).The obtained optimal parameters of the extraction procedure of Opuntia Milpa Alta polysaccharides were: water to solid ratio 20:1, time 2 h, temperature 50℃, and the average extract content of polysaccharides 17.6 mg/mL.Ion exchange resins(IER)were used to desalinize and decolorize Opuntia Milpa Alta powder extract. 001×7-D309 ion exchange combination resins were screened from 12 pairs of different resins based on the static adsorption results which the desalination and the decolorization rates were about 92.0% and 78.6% respectively after treatment by 001×7-D309. In further dynamic adsorption study,the optimal flow rate of Opuntia Milpa Alta powder extract treated with 001×7-D309 was 4 BV·h^-1.When the sample was 7-fold resin volume, the desalination and the decolorization rates reached 96.0% and 80.0%, at the same time about 0.7% soluble solids were lost by 001×7 and D309 combination resins absorption. The desalination and decolorization rates of 001×7-D309 ion exchange combination resins can be stabilized at 95.0⁹6.0% and 80.0% during 7 times'recycling usage, which indicated this combination resins(001×7 and D309 )had good regeneration performance. T-G system was first found it had a decreasing effect on the solution conductivity, and a good linear relationship was presented. 10% T can decrease the solution conductance to 63% while 10% T and 23% G combination can reduce conductance to 32%.7 kinds of cosmetics stability tests indicated that 10% T and 23% G combination system can improve the stability of Opuntia Milpa Alta powder extract liquid.The experimental anti-allergy and anti-irritation compound solution had good anti-allergic and free radicals clearance effects. Under 2.0% dosage, the hyaluronic acid enzyme inhibition rate, O₂, OH and DPPH radicals clearance rates were 87.3%, 76.0%, 68.1% and 17.0%, respectively. Within 35 minutes UV irradiation, RBC hemolysis time of RBC suspension added 5.0% compound solution occurred slowly than normal suspension. ELLSA test results indicate that vascular endothelial cells growth factor receptor Flt1 inhibition rate reached 35.4% with 5% compound solution usage, showing that the anti-allergy and anti-irritation compound solution had a certain preventive effects of UV-damage cells and facial telangiectasia.RBC haemolysis test model and Comet assay were used to determine the protective effects of anti-allergy and anti-irritation compound solution on red blood cell membrane and mouse hepatocyte DNA damage induced by sodium dodecyl sulfate (SDS).The results showed that 20¹00μg/mL anti-allergy compound solution had no damage to cell membrane and DNA.When the dosage of 40⁸0μg/mL, the anti-allergy compound solution can prevent cell membrane and DNA damage induced by SDS, and the dosage-response relationships were obvious. Through fluorescence microscope observations, it was found that anti-allergy compound solution showed the ability of preventing DNA damage significantly when the concentration is greater than 60μg/mL. In the 100μg/mL of use, the comet-like cell head and tail gathered together and cell DNA damaged and tail length trailing rate decreased 58% and 57.8%. Anti-allergy compound solution can inhibit the growth of Fecal coliforms and Staphylococcus aureus in logarithmic growth phase. Observation under the transmission electron microscope revealed that the microbe'growth, reproduction and metabolism are inhibited after effected 10h by anti-allergy compound solution. And the compound solution destroyed the structure of the microbe, made the celI distorted, then the cell walls was broken that the cytoplasm flow out, until it became empty or was decomposed into granule residue, which formed the opposite effect compared to the protection of animal red blood cells. Human efficacy evaluation showed that anti-allergy compound solution can calm the skin symptoms immediately with the effective rate 86.9%; The anti-allergy compound solution can also enhance skin tolerance significantly; Within 60 min, the cream added 5.0% anti-allergy compound solution can decrease skin redness and swelling 52.7%; Capsaicin induced itch and then applicated by the cream 10 min, the itching decreased 77.8%. Clinical efficacy evaluation results show that the patients'skin sensitivity, itching and red swelling degree were improved after application of the cream added 5.0% anti-allergy compound solution. The total effective rate of experimental group were 93.6%, 95.5% and 91.3%, respectively. And 97.8% patients considered the safety and tolerability of the cream as "very good" or "good."...