Evaluation of Physicochemical and Microbial Properties of Extracts from Wine Lees Waste of Matelica’s Verdicchio and Their Applications in Novel Cosmetic Products
Abstract
:1. Introduction
2. Materials and Methods
2.1. Wine Lees
2.2. Reagents and Materials Other Than Wine Lees
2.3. Microbiological Analysis of Native Wine Lees
2.3.1. Extraction of Total DNA
2.3.2. Quality Check by PCR and Agarose Gel Electrophoresis
2.3.3. Preparation and Shipment of Samples
2.3.4. Preparation and Inoculation of Microbial Growth Media
2.3.5. Bacterial Population Enrichment
2.3.6. Microbial Growth on Selective and Differential Media
2.3.7. DNA Extraction by GenElute Bacterial Genomic DNA Kit
2.3.8. Quality Check by PCR and Agarose Gel Electrophoresis
2.3.9. Preparation and Shipment of Samples for NGS Analysis
2.3.10. NGS Bioinformatic Analysis Using DADA2
2.4. Ultrasound-Assisted Yeast Lysis of NWLs and Physicochemical Characterization of the Aqueous Extract
2.4.1. Ultrasound-Assisted Yeast Lysis of NWLs
2.4.2. Scanning Electron Microscopy (SEM)
2.4.3. Dynamic Light Scattering (DLS)
2.4.4. Bio-Rad Protein Assay
2.4.5. Quantitative Element Analysis
2.4.6. Total Phenol Content (Folin-Ciocalteu Assay)
2.4.7. Antioxidant Assays
DPPH Radical Scavenging Method
ABTS Radical Scavenging Method
Ferric Reducing Antioxidant Power (FRAP)
2.4.8. Quantitation of Squalene by High-Performance Liquid Chromatography Coupled with Diode Array Detection
2.5. Determination of Cell Metabolic Activity and Cytotoxicity
2.5.1. Cell Culture and Treatments
2.5.2. Cell Metabolic Activity Test (MTT)
2.5.3. Cytotoxicity Assay
2.6. Statistics
2.7. Formulation of Cosmetic Products
2.8. Physicochemical Characterization of Formulations
2.9. Challenge Test
2.10. Local Compatibility Test with Human Skin (Irritant Potential)
2.11. Measurement of the Trans Epidermal Water Loss (TEWL)
2.12. Measurement of the Sebometry
3. Results and Discussions
3.1. Microbiological Analysis
3.1.1. Analysis of the Microbiome in the Lees
3.1.2. Count of Mesophilic Bacteria, Anaerobes, and Fungi
3.1.3. Growth of Isolates on Selective Media
3.1.4. Identification of Mesophiles Isolates Grown onto MSA by 16S rDNA Analysis
3.2. Optimization of Ultrasound-Assisted Extraction Method for the Yeast Cells Breakdown in Water and Evaluation of the Properties of the Aqueous Extract
3.3. In Vitro Metabolic Activity and Cytotoxicity Assays
3.4. Cosmetic Products
3.4.1. General Characteristics of the Cosmetic Products
3.4.2. Evaluation of Clinical Efficacy
3.4.3. Measurements of the Sebometry
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Medium | Sample | Incubation Temperature (°C) | Atmospheric Conditions |
---|---|---|---|
TSA | F1:10 | 37 | Aerobiosis |
TSA | F1:100 | 37 | Aerobiosis |
TSA | F1:10 | Room temperature | Aerobiosis |
TSA | F1:100 | Room temperature | Aerobiosis |
SDA | F1:10 | 37 | Aerobiosis |
SDA | F1:100 | 37 | Aerobiosis |
SDA + chloramphenicol | F1:10 | 37 | Aerobiosis |
SDA+ chloramphenicol | F1:100 | 37 | Aerobiosis |
MRS | F1:10 | Room temperature | Aerobiosis |
MRS | F1:100 | Room temperature | Aerobiosis |
MRS | F1:10 | Room temperature | Anaerobiosis |
MRS | F1:100 | Room temperature | Anaerobiosis |
Step | 1 | 2 | 3 |
---|---|---|---|
T (°C) | 150 | 190 | 50 |
P (Bar) | 36 | 36 | 0 |
POWER (%) | 70 | 90 | 0 |
RAMP (Min) | 5 | 5 | 1 |
Step (Min) | 10 | 20 | 20 |
Commercial name | Ingredients (INCI) | Supplier |
---|---|---|
TEGO® Betain CK D MB | Cocamidopropyl BetaineSodium Chloride, Aqua | Evonik, Essen, Germany |
Stearic acid | Acef, Fiorenzuola d’Arda (PC), Italy | |
Cetearyl alcol | ||
Theobroma Cacao Seed Butter | ||
Butyrospermum Parkii Butter | ||
Citric acid | ||
Glycerin | ||
Kaolin | ||
Zinc oxide | ||
Helianthus Annuus Seed Oil | ||
Cera Alba | ||
Hydrogenated castor oil | ||
Oryza Sativa Starch | ||
Argania Spinosa Kernel Oil | ||
Oryza Sativa Bran Oil | ||
ACNIBIO PE 9010 | Phenoxyethanol, ethylhexylglycerin | |
GLDA Chelating | Aqua, Tetrasodium glutamate diacetate, Sodium hydroxide | |
Butylene Glycol | ||
Oryza Sativa Bran Oil | ||
Sodium Stearate | ||
Thocopheryl Acetate | Basf, Ludwigshafen, Germany | |
Sodium Coco-Sulfate | ||
Glyceryl Stearate | ||
Betaine | ||
Erylite | Eurotrading, Civitanova Marche (MC), Italy | |
Cocos nucifera oil | Esperis, Milan, Italy | |
Copernicia Cerifera Cera | Natura Tec, Louis Lépine Fréjus, France | |
Turkey Red Oil | Castor Oil Sulfated, Sodium Salt, Aqua | Zschimmer & Schwarz, Lahnstein, Germany |
PROTELAN ENS | Stearic acid, Sodium lauroyl glutamate, Cetearyl alcohol, Glyceryl stearate | |
Hydrogenated Coco-glycerides | Farmalabor, Canosa di Puglia (BT), Italy | |
Polyglyceryl-10 Caprylate | Bregaglio, Biassono (MB), Italy | |
SORBOSIL™ AC 36 | Hydrated silica, Aqua | |
Cetyl Alcohol | ||
Sodium Cocoyl Glutamate | Prodotti Gianni, Milan, Italy | |
Hydroxyethylcellulose | Vevy Europe, Genova, Italy |
Cosmetic Formulation/ Consistency | List of Ingredients According to the International Nomenclature Cosmetic Ingredients (INCI) | Method of Preparation |
---|---|---|
Facial cleanser Solid | Sodium Coco-sulfate, Oryza Sativa Starch, Sodium Cocoyl Glutamate, Cetearyl Alcohol, Glyceryl Stearate, Hydrogenated Coco-glycerides, Saccharomyces/Grape Lees Ferment Extract, Butyrospermum Parkii Butter, Betaine, Hydrogenated Coco—glycerides, Aqua, Theobroma Cacao Seed Butter, Tocopheryl Acetate, Tocopherol, Parfum, Citric Acid | The lipidic phase A was weighed and heated in a water bath up to 85 °C until complete melting. Vitamin E was added to the Phase A once the latter was removed from the water bath and allowed to cool under stirring. Sodium coco-sulfate, oryza sativa starch and the Saccharomyces/Grape Lees Ferment were weighed and homogenized to the lipidic phase at 3500 rpm to obtain a pasty texture. Citric acid was weighed and stirred until complete solubilization in water and added to the paste to reach the desired pH. Perfume was added as the last step. |
Facial cleansing Powder | Oryza Sativa Starch, Saccharomyces/Grape Lees Ferment Extract, Oryza Sativa Bran Oil, Sodium Cocoyl Glutamate, Erylite, Silica, Betaine, Cocamidopropyl Betaine, Sodium Chloride, Parfum, Aqua | The oil was added drop by drop to be adsorbed into the weighed starch by using a pestle and a mortar. Thus, first Saccharomyces/Grape Lees Ferment, and then betaine, silica and erylite were added and mixed with a mortar to obtain a homogeneous powder. Perfume was added as the last step. |
Make-up remover Solid | Helianthus Annuus Seed Oil, Hydrogenated Coco-glycerides, Cera Alba, Polyglyceryl-10 Caprylate, Saccharomyces/Grape Lees FermentExtract, Theobroma Cacao Seed Butter, Castor Oil Sulfated, Cocos Nucifera Oil, Butyrospermum Parkii Butter, Hydrogenated Castor Oil, Sodium Salt, Tocopheryl Acetate, Parfum, Aqua. | The lipidic phase was weighed and heated in a water bath up to 90 °C until complete melting. Vitamin E was added after the removal from the water bath of the mixture and allowed to cool under stirring. Saccharomyces/Grape Lees Ferment was dispersed in water and added to sulfated castor oil s, heated up to 40 °C under stirring to promote the formation of the mixture and added to the lipidic phase. Perfume was added as the last step. |
Melting face mask Solid | Cocos Nucifera Oil, Kaolin, Saccharomyces/Grape Lees FermentExtract, Butyrospermum Parkii Butter, Copernicia Cerifera Cera, Castor Oil Sulfated, Sodium Salt, Zinc Oxide, Tocopheryl acetate, Parfum, Aqua. | Cocos nucifera oil, butyrospermum parkii butter and cera carnauba were weighed and heated up to 80 °C for 30 min under stirring with kaolin and zinc oxide. After removing the ingredients from the water bath, vitamin E was added. Saccharomyces/Grape Lees Ferment was dispersed in water and added to sulfated castor oil, heated up to 40 °C under stirring to promote the formation of the mixture and added to the other ingredients. Perfume was added as the last step. |
Purifying face mask Solid | Aqua, Kaolin, Glycerin, Saccharomyces/Grape Lees FermentExtract, Butyrospermum Parkii Butter, Sodium Stearate, Butylene Glycol, Glyceryl Stearate, Cetearyl alcohol, Cera Alba, Stearic Acid, Sodium Lauroyl Glutamate, Tocopheryl Acetate, Phenoxyethanol, Hydroxyethylcellulose, Parfum, Ethylhexylglycerin, Tetrasodium Glutamate Diacetate, Sodium Hydroxide. | Phase A, composed of water, chelating and wetting agents, kaolin, sodium stearate and Saccharomyces/Grape Lees Ferment, was weighed and heated up to 75 °C under stirring. Hydroxyethylcellulose was then slowly added to the phase A under stirring. Phase B, composed of the lipidic ingredients, was heated up to 75 °C and added to the phase A under homogenization (3500 rpm) until the temperature reached 40 °C. Vitamin E, preservative and parfum were added as the last step. |
Bacterial Genus | Temperature Range(°C) | pH Range | Salt Range (%) | Oxygen | Mannitol Fermentation | References |
---|---|---|---|---|---|---|
Bacillus | 4–49 | 6–9 | 0–10 | aerobic | + | [58] |
Lysinibacillus | 10–45 | 5.5–9.5 | 5–7 | aerobic | − | [59] |
Oceanobacillus | 10–40 | 6.5–10 | 3–10 | aerobic | +/− | [60] |
SAMPLES | FREEZE-DRYING | SONICATION | SIZE | PDI |
---|---|---|---|---|
NWL0 | − | − | 2945 ± 26 | 0.084 |
SWL0 | − | + | 1476 ± 64 | 0.442 |
NWL1 | + | − | 2447 ± 28 | 0.676 |
SWL1 | + | + | 1723 ± 35 | 0.291 |
ELEMENT ANALYSIS | |||
---|---|---|---|
NWLs (ppm) | Supernatant of the NWLs (ppm) | Supernatant of the SWLs (ppm) | |
Li | 0.06 ± 20.29 | 0.20 ± 6.41 | 47.26 ± 3.12 |
B | 58.18 ± 1.11 | 168.94 ± 1.35 | 236.22 ± 1.90 |
Na | 68.98 ± 2.91 | 315.89 ± 1.18 | 729.07 ± 4.23 |
Mg | 577.88 ± 2.07 | 1735.23 ± 1.55 | 2125.98 ± 2.94 |
P | 10,888.45 ± 1.56 | 9146.64 ± 1.27 | 7862.77 ± 2.56 |
S | 3593.28 ± 3.11 | 4928.72 ± 1.78 | 5383.26 ± 5.85 |
K | 13,514.31 ± 1.74 | 19,195.52 1.68± | 65,338.26 ± 3.23 |
Ca | 2687.06 ± 1.45 | 1130.35 ± 1.34 | 7927.05 ± 2.49 |
Cr | 0.43 ± 2.15 | 1.10 ± 1.12 | 1.39 ± 3.41 |
Mn | 10.72 ± 1.13 | 8.48 ± 1.40 | 20.63 ± 2.80 |
Fe | 11.48 ± 1.92 | 8.28 ± 2.03 | 4.49 ± 6.68 |
Cu | 227.34 ± 1.74 | 588.19 ± 0.91 | 262.41 ± 2.67 |
Zn | 5.16 ± 4.94 | 14.46 ± 2.47 | 45.54 ± 2.25 |
Rb | 12.21 ± 1.14 | 35.03 ± 1.72 | 62.21 ± 2.80 |
Sr | 9.11 ± 1.81 | 11.93 ± 0.70 | 25.66 ± 1.62 |
Ba | 2.16 ± 1.25 | 3.25 ± 1.12 | 10.53 ± 1.85 |
Hg | 0.00 ± 6.49 | 0.02 ± 5.91 | 0.00 ± 14.57 |
ABTS | DPPH | FOLIN | FRAP | |||
---|---|---|---|---|---|---|
IC50 (mg/mL) | μmol TE/g | IC50 (mg/mL) | μmol TE/g | mg GAE/g | mg TEA/g | |
NWL | 0.107 ± 0.003 | 204.525 ± 4.756 | 0.457 ± 0.009 | 870.692 ± 17.961 | 81.333 ± 9.905 | 55.048 ± 6.419 |
SWL | 0.068 ± 0.001 | 182.489 ± 2.643 | 0.228 ± 0.020 | 607.195 ± 64.368 | 136.533± 8.942 | 82.133 ± 8.987 |
TROLOX | 0.004 ± 0.000 | 7.991 ± 0.190 | 0.003± 0.000 | 120.577 ± 14.872 |
Cosmetic Formulation | pH | Physicochemical Stability | Challenge Test | Mean Irritation Index | ||
---|---|---|---|---|---|---|
Accelerated Stability | Long Term Stability | 15 min | 24 h | |||
Solid facial cleanser | 5.85 | Stable | Stable | Passed | 0 | 0 |
Facial cleansing powder | 5.84 | Stable | Stable | Passed | 0 | 0 |
Make-up remover | 6.08 | Stable | Stable | Passed | 0 | 0 |
Melting mask | 6.05 | Stable | Stable | Passed | 0 | 0 |
Purifying mask | 5.80 | Stable | Stable | Passed | 0 | 0 |
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© 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Di Nicolantonio, L.; Ferrati, M.; Cristino, M.; Peregrina, D.V.; Zannotti, M.; Vitali, L.A.; Ciancia, S.I.; Giovannetti, R.; Ferraro, S.; Zara, S.; et al. Evaluation of Physicochemical and Microbial Properties of Extracts from Wine Lees Waste of Matelica’s Verdicchio and Their Applications in Novel Cosmetic Products. Antioxidants 2023, 12, 816. https://doi.org/10.3390/antiox12040816
Di Nicolantonio L, Ferrati M, Cristino M, Peregrina DV, Zannotti M, Vitali LA, Ciancia SI, Giovannetti R, Ferraro S, Zara S, et al. Evaluation of Physicochemical and Microbial Properties of Extracts from Wine Lees Waste of Matelica’s Verdicchio and Their Applications in Novel Cosmetic Products. Antioxidants. 2023; 12(4):816. https://doi.org/10.3390/antiox12040816
Chicago/Turabian StyleDi Nicolantonio, Lucrezia, Marta Ferrati, Maria Cristino, Dolores Vargas Peregrina, Marco Zannotti, Luca Agostino Vitali, Sonia Ilaria Ciancia, Rita Giovannetti, Stefano Ferraro, Susi Zara, and et al. 2023. "Evaluation of Physicochemical and Microbial Properties of Extracts from Wine Lees Waste of Matelica’s Verdicchio and Their Applications in Novel Cosmetic Products" Antioxidants 12, no. 4: 816. https://doi.org/10.3390/antiox12040816