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Purpose: This study aimed to determine the optimal blending ratio of extracts of Hippophae rhamnoides (H. rhamnoides) fruit, Rubus fruticosus (R. fruticosus) leaf, and Perillae folium (P. folium) leaf as a potential functional material for antiwrinkle, antioxidant, anti-inflammatory, and whitening properties. Methods: We used the design of experiment (DOE) efficacy method to identify the optimal blend ratio for a natural extract complex. In addition, we tested the efficacy of the extract complex and the individual extracts by experiments, including elastase inhibition assay, NO inhibition assay, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity assay, tyrosinase inhibition assay, and ROS inhibition assay. Results: DOE methods using natural extracts showed that the most efficacious ratio of H. rhamnoides fruit: R. fruticosus leaf: P. folium leaf was 0.25:0.7:0.05. At this ratio, the elastase inhibitory effect, an indicator of wrinkle improvement, was 28% compared to the control. The antioxidant activity was 71.8% higher than the control. Additionally, NO formation was significantly suppressed by the extract complex. Tyrosinase inhibitory activity, a whitening index, showed 24.4% improvement over the control. Finally, hydrogen peroxide-induced cytotoxicity was significantly inhibited following treatment with the extract complex. Conclusion: The natural extract complex of H. rhamnoides, R. Fruticosus, and P. folium was found to have beneficial wrinkle improvement, antioxidant, anti-inflammatory, and whitening effects, and has potential as a functional cosmetic material.

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In this study, we aim to study the functionality of multiple effects of peptide collagen and naturally derived compounds through in vitro experiments and human application tests. Method: Cytotoxicity of inner beauty film (IBF), pro-collagen generation ability, elastase and collagenase inhibition, DPPH radical scavenging and tyrosinase inhibition ability were measured, and skin improvement, through in vitro and human application test. Results: Cytotoxicity was not confirmed up to 1000 μg/mL. Elastase inhibition was 7.2% at a concentration of 125 μg/mL, and collagenase showed an inhibition rate of 16.3% at 31.3 μg/mL. The pro-collagen synthesis showed expression of 34%, compared with the negative control when the concentration was 62.5 μg/mL. The antioxidant activity of 250 μg/mL and whitening effect of 62.5 μg/mL are showed in the Results section, and there was no observed deformation or color change even after confirming the stability for 6 months. Even in the human clinical study, the wrinkle improvement before and after using the IBF was 32.2% at maximum, skin melanin and brown spot improvement was 32.5%, pore improvement was 31.5%, and skin density improvement was 12.8%, as shown in the Results section. Conclusion: Therefore, the IBF containing the complex of peptide collagen and naturally derived compounds has a positive effect on wrinkle improvement, skin melanin and brown spot improvement, skin pore improvement, and skin density improvement, as well as exhibiting antioxidant and whitening effects. As a result, IBF can be developed as a key material for cosmetics and foods.