Method for evaluating anti-aging effect of skin by combining multiple in-vitro models based on enzymology system
A model evaluation and anti-aging technology, which is applied in biochemical equipment and methods, scientific instruments, microbial measurement/inspection, etc., can solve problems such as high price, inability to realize large-scale application, irregular expression of markers, etc.
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Embodiment 1
[0071] Example 1 Evaluation of the anti-aging effect of a certain brand of glucosyl rutin based on chemical level, cellular level, and 3D skin model
[0072] 1. Prediction of anti-aging efficacy of raw materials based on chemical level
[0073] Elastase inhibition test: 100 μL of 0.2 mol / L tris-HCl buffer (Ph=8.0), 25 μL of 10 mmol / L MAAPVN (N-methoxysuccinyl-alanine-alanine-proline Amino acid-valine-4-nitroaniline) and 50 μL of raw material dilution were mixed, and incubated at 25°C for 15 minutes; then 25 μL of 0.3U / mL elastase (the highest activity) was added to continue incubation for 15 minutes. The microplate reader detects the OD value under the condition of 410nm, and calculates the inhibition rate by the following formula. EGCG ((Epigallocatechin gallate, epigallocatechin gallate) is a positive control. Inhibition rate (%)=[(absorbance of control group-absorbance of sample group) / absorbance of control group]×100%. Different concentrations of test substances There is...
Embodiment 2
[0106] Example 2 Evaluation of anti-aging effects of a certain brand of tocopherol derivatives based on chemical level, cellular level, and 3D skin model methods
[0107] 1. Prediction of anti-aging efficacy of raw materials based on chemical level
[0108] Hyaluronidase inhibition test: Mix 100 μL of hyaluronidase, 100 μL of PBS buffer, 50 mmol / L of NaCl and 0.01% BSA with 50 μL of sample solution as a medium, and incubate in the medium at 37°C for 10 min . Then add 100 μL of hyaluronidase solution (0.03%, 300 mmol / L phosphate solution, pH=5.35) as the start of reaction, and incubate at 37° C. for 45 min. Unhydrolyzed hyaluronic acid was precipitated with 1.0 mL of acid albumin solution (0.1% BSA solution 24 mmol / L sodium acetate and 79 mmol / L acetic acid, pH=3.75). Then the mixture was left at room temperature for 10 min, and the absorbance of the mixture was detected at 600 nm with a microplate reader. The test group without enzyme was the control of the maximum inhibiti...
Embodiment 3
[0142] Example 3 Evaluation of the anti-aging effect of a certain brand of collagen derivatives based on chemical level, cellular level, and 3D skin model methods
[0143] 1. Prediction of anti-aging efficacy of raw materials based on chemical level
[0144] 1.1 Collagenase inhibition test: Use ninhydrin reaction to monitor the amount of peptides fragmented by collagenase, mix collagen I with TES (pH7.5, containing Tris-HCl, EDTA and SDS) buffer, dilute The test substance and collagen IV were mixed evenly, incubated at 37°C for 5h, and then centrifuged at 2000rpm for 5min. Collect the supernatant, mix it with ninhydrin, heat at 80°C for 10min and cool down. The solution was mixed with isopropanol at a ratio of 1:1, and centrifuged at 12000 rpm for 10 min at 4 °C. The supernatant was added to a 96-well plate, and its absorbance was detected at 600 nm. Calculate inhibition rate. Take EGCG ((Epigallocatechin gallate, epigallocatechin gallate) as positive control. Inhibition r...
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