74 UPCYCLED INGREDIENTS
■ Physical characterization: colour and appearance are evaluated as key parameters to be considered for applications in the cosmetic industry.
In vitro studies for cosmetics applications of early-stage apple extracts ■ Antioxidant capacity by ABTS determination: Cell free in vitro test for the evaluation of chemical antioxidative capacity. The assay principle is based on the reductive decolourization of the green coloured cationic radical 2,2’-Azinobis (3-ethylbenzothiazoline 6-sulfonic acid) by antioxidants. The decolourization can be measured spectrophotometrically at 734 nm. ■ Determination of Radical Oxygen Species (ROS): Fibroblasts are treated with cumene as the positive control to stimulate ROS production. Fibroblast are treated as well with cumene in the presence of different concentrations of early-stage apple extract. ROS are determined by fluorescence technique. ■ Hyaluronic Acid Determination: The analytical method utilizes the ELISA technique measuring the quantity of hyaluronic acid in cell lysates based on specific antibodies coupled with enzymes catalyzing chromophore formation. The chromophore absorbance is proportional to the quantity of hyaluronic acid in the sample.
Results Extraction Table 2 shows the yields obtained after each proposed extraction. These yields are based on laboratory scale extraction and may differ from the yields that can be achieved for industrial batches. The results show a similar performance on
the extraction independently of the system and ratio of solvent used.
Characterization of standardized early-stage apple extracts Characterization process takes place in standardized early-stage apple extracts: A1, B1 and C1, to guarantee that the dry matter content on each formulation is the same, independently of the extraction system, and thereby to run a fair comparison on their concentration of molecules of interest. Table 3 and Figures 3 and 4 show the results that are obtained. According to these results, trial C1 is the
Figure 2: Recovered early-stage apples
TABLE 1: CONDITIONS APPLIED DURING HYPERFREQUENCY APPLICATION TO EARLY-STAGE APPLES Phase A B
Solvent (kg) 100% water C
50% water, 50% ethanol 67% water, 33% ethanol
Total Solvent (kg) 4 4
6
TABLE 2: EXTRACTION YIELD ON DRY MATERIAL Phase
A B C
Extract yield on dry material 14.3%
13.2% 10.8%
most interesting to meet cosmetic applications, considering that in its appearance, this extract does not present any signs that can lead to sedimentation problems in the future. Moreover, C1 trial has a higher concentration of Phlorizin than trials A1 and B1. Considering these results, the following
phase of the study, in vitro studies trial C1, is the chosen extract to be evaluated.
In vitro studies for cosmetics applications of early-stage apple extract C1 Apples represent a natural source of compounds with a wide range of applications in the cosmetic industry. Polyphenols and fruit acids have been identified in different varieties of fruit and these components have been
TABLE 3: COMPOSITION AND CHARACTERIZATION OF EARLY-STAGE APPLE EXTRACTS Formulation
Molecules of interest Phlorizin A1 B1 C1 240ppm 230ppm 265ppm Chlorogenic Acid 500ppm 410ppm 480ppm PERSONAL CARE September 2023 Appearance
Physical Characterization Colour
Cloudy liquid with deposits Yellow pale brown
Cloudy liquid with deposits
Clear liquid
Yellow brown
Yellow Orange
Figure 3: Physical appearance formulations A1, B1 and C1
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found to have a positive impact to improve the skin, driven mainly by its antioxidant and moisturization properties. As it has been previously explained,
early-stage apples are reported to have a higher concentration in specific polyphenols, especially, chlorogenic acid and phlorizin. ■ Chlorogenic acid (CGA): It is a phenolic compound commonly found in coffee and green tea extracts, among other natural ingredients, like apples. CGA content is higher in early-stage apples compared with mature apples. Studies have shown the antioxidant, antibacterial and anti-inflammatory capacity of CGA.3,4,5 ■ Phlorizin: It is a glucoside of phloretin which is naturally found in unripe/early-stage apples. Studies show the potential antioxidant and protection of skin barrier effects of Phlorizin.6,7 Therefore, early-stage apple extract C1, has been tested to evaluate its antioxidant capacity (ABTS and ROS) and its moisturization potential (supporting the Hyaluronic acid modulation), as Figures 5, 6 and 7 show respectively.
Mass Raw Material
Time Constant values for each trial
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