Transforming The Face Of Preservation: Peptide Technology In The Personal Care Industry Antimicrobials A Natural Approach to Product Preservation

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In The Personal Care Industry Antimicrobials A Natural Approach to Product Preservation Anna Crovetto - Technical Marketing - Email: a.crovetto@activeconcepts.it

Contents 1. Preservation: Chemist vs. Consumer 2. Changes within the Personal Care Market 3. The Antimicrobial Market 4. Active Micro Technologies Peptide Systems 5. M15008-Leucidal Liquid 6. M15019-Leucidal Liquid SF 7. Summary

1. Preservation: The Chemist Parabens: the most frequently used preservatives in cosmetic and beauty care products Effective Low use levels Safety data History of use Low cost Low irritation potential Synthetic Negative press

1. Preservation: The Consumer Demonisation of parabens and synthetic preservatives in the media Instigated by a research paper from the University of Reading Darbre PD. Et al (2004) Concentrations of parabens in human breast tumours, [1] Esters of p-hydroxybenzoic acid (parabens) detected in breast tumour samples Low threshold study: Limited test group (20 breast tumour samples) No control samples No male samples Evidence did not imply that parabens cause abnormalities

[2] Media Reaction Sensationalism Mis-Quote Demonisation

2. Changes within the Personal Care Market March 2011 Denmark enforces a ban on propyl and butyl paraben, their isoforms and their salts in cosmetic products for children up to three years of age. [3] May 2012 The French Agence Nationale de Sécurité du Médicament (ANSM) questions the safety of Phenoxyethanol for young children. [4] July 2013 The British Association of Dermatologists (BAD) presents research highlighting an epidemic of contact dermatitis due to two synthetic preservatives, Methylisothiazolinone and Methylchloroisothiazolinone / Methylisothiazolinone. [5] October 2013 The European Commission calls for all data on Phenoxyethanol and o-phenylphenol. Their purpose is to gather all scientific information in preparation for the Scientific Committee for Consumer Products (SCCS), who will publish its opinions on these two ingredients. [4] December 2013 - The personal care association, issues an industry-wide recommendation to discontinue the use of the preservative Methylisothiazolinone (MIT) in leave-on skin cosmetics and personal care products. [6]

3. The Antimicrobial Market Alcohols Example: Benzyl Alcohol Organic Acids / Salts Example: Potassium Sorbate Multifunctional Additives Example: Caprylyl Glycol Natural Flavours and Fragrance Example: Parfum (p-anisic Acid)

3. The Antimicrobial Market

4. Active Micro Technologies Peptide System ACTIVE MICRO TECHNOLOGIES (AMT) FOCUS: To identify and develop unique antimicrobial ingredients GOAL: To provide antimicrobials that are effective, safe, and stable INNOVATION: - cosmetic active benefits - restricting microbial activity

4. Active Micro Technologies Peptide System Lactic Acid Bacteria (LAB) Family Produce novel antimicrobial peptides Bacteriocins Proteinaceous materials Inhibit growth of bacterial strains Structurally, functionally, and ecologically diverse

5. M15008-Leucidal Liquid INCI: Leuconostoc/Radish Root Ferment Filtrate The next generation of proactive, innovative and efficacious antimicrobial systems

5. M15008-Leucidal Liquid Mechanism of Action LAB family Leuconostoc kimchii produces lactic acid Restricts the growth of microorganisms by acidifying their environment Fermentation of Leuconostoc in a defined growth medium containing Radish creates bacteriocins Provides broad spectrum antimicrobial activity Proven conditioning benefits Modulated Activity Willow Bark Extract is added facilitates controlled cell lysis Ensures the release of the antimicrobial peptides for maximised activity

5. M15008-Leucidal Liquid Challenge Test in a Cream Formulation MIC Data MIC % E. coli 1.60 S. aureus 0.80 P. aeruginosa 1.60 C. albicans 0.80 A. brasiliensis 2.40 Table 1: Challenge Test Results for 2.00% M15008-Leucidal Liquid in O/W Cream formulation inoculated on day 0 and re-inoculated on day 28

5. M15008-Leucidal Liquid Compatibility with Water + Glycerin+ Carbopol Ultrez 20 + NaOH (ph 7)

5. M15008-Leucidal Liquid Product Code: M15008 INCI: Water + Leuconostoc/Radish Root Ferment Filtrate INCI Status: Conforms Suggested Use Level: 2.00-4.00% Suggested Applications: Skin + Hair Care, Moisturising, Conditioning, Anti-Microbial

6. M15019-Leucidal Liquid SF INCI: Lactobacillus Ferment The next generation of proactive, innovative and efficacious antimicrobial systems

6. M15019-Leucidal Liquid SF Mechanism of Action LAB family Lactobacillus bulgaricus produces lactic acid Restricts the growth of microorganisms by acidifying their environment Fermentation of Lactobacillus creates bacteriocins Provides broad spectrum antimicrobial activity Proven conditioning benefits Modulated Activity Lysozyme is added to the ferment filtrate facilitates controlled cell lysis Ensures the release of the antimicrobial peptides for maximised activity

6. M15019-Leucidal Liquid SF Challenge Test in a Cream Formulation MIC Data MIC % E. coli 0.75 S. aureus 1.00 P. aeruginosa 1.00 C. albicans 0.75 A. brasiliensis 1.00 Table 2: Challenge Test Results for 2.0% M15019-Leucidal Liquid SF in O/W Cream formulation inoculated on day 0 and re-inoculated on day 28

6. M15019-Leucidal Liquid SF PCPC Method 3 Determination of Preservation Adequacy of Water Miscible Cosmetic and Toiletry Formulations Generic Base Cream + 2.0% M15019-Leucidal Liquid SF ph 3, 4, 5, 6, 7, 8, and 9 Purpose These studies were initiated to determine the efficacy of a cosmetic ingredient with antimicrobial properties in cream formulations against bioburden as a function of time. Tested Organisms Escherichia coli Pseudomonas aeruginosa Staphylococcus aureus Aspergillus brasiliensis

6. M15019-Leucidal Liquid SF Challenge Test in Generic Base Cream ph 3 Table 3: Challenge Test Results for Generic Cream with 2.0% M15019-Leucidal Liquid SF inoculated on Day 0. Results show percent (%) reduction in viable organisms. The results of this Challenge Test demonstrate the effectiveness of Leucidal Liquid SF as an antimicrobial agent in a cream formulation ph 3. The gram negative and gram positive bacteria were reduced by greater that 99.9% within 7 days of each challenge. The mould was reduced by greater than 90.0% within 7 days of each challenge.

6. M15019-Leucidal Liquid SF Challenge Test in Generic Base Cream ph 5 The results of this Challenge Test demonstrate the effectiveness of Leucidal Liquid SF as an antimicrobial agent in a cream formulation ph 5. The gram negative and gram positive bacteria were reduced by greater that 99.9% within 7 days of each challenge. The mould was reduced by greater than 90.0% within 7 days of each challenge. Table 5: Challenge Test Results for Generic Cream with 2.0% M15019-Leucidal Liquid SF inoculated on Day 0. Results show percent (%) reduction in viable organisms.

6. M15019-Leucidal Liquid SF Challenge Test in Generic Base Cream ph 7 The results of this Challenge Test demonstrate the effectiveness of Leucidal Liquid SF as an antimicrobial agent in a cream formulation ph 7. The gram negative and gram positive bacteria were reduced by greater that 99.9% within 7 days of each challenge. The mould was reduced by greater than 90.0% within 7 days of each challenge. Table 7: Challenge Test Results for Generic Cream with 2.0% M15019-Leucidal Liquid SF inoculated on Day 0. Results show percent (%) reduction in viable organisms.

6. M15019-Leucidal Liquid SF Challenge Test in Generic Base Cream ph 9 The results of this Challenge Test demonstrate the effectiveness of Leucidal Liquid SF as an antimicrobial agent in a cream formulation ph 9. The gram negative and gram positive bacteria were reduced by greater that 99.9% within 7 days of each challenge. The mould was reduced by greater than 90.0% within 7 days of each challenge. Table 9: Challenge Test Results for Generic Cream with 2.0% M15019-Leucidal Liquid SF inoculated on Day 0. Results show percent (%) reduction in viable organisms.

6. M15019-Leucidal Liquid SF In-vivo Moisturisation Assay Table 10: Change in moisturisation comparing the control and 1.0% M15019-Leucidal Liquid SF + Base Cream (4 week study).

6. M15019-Leucidal Liquid SF Product Code: M15019 INCI: Lactobacillus Ferment INCI Status: Conforms Suggested Use Level: 2.00-4.00% Suggested Applications: Skin + Hair Care, Moisturising, Conditioning, Anti-Microbial

Summary: The cosmetics industry is moving away from synthetic preservative systems Driven by consumer demand, media exploitation and lack of understanding Active Micro Technologies can provide alternative natural antimicrobials Leucidal Liquid and Leucidal Liquid SF: Detailed and substantiated efficacy data Thorough safety data Multifunctional benefits

Bibliography: 1. Darbre PD, Aljarrah A, Miller WR, Coldham NG, Sauer MJ, Pope GS.(2004) Concentrations of parabens in human breast tumours, [online] Available: http://www.ncbi.nlm.nih.gov/pubmed/14745841 [09.10.13] 2. Bates C. (2012) Chemical found in deodorants, face cream and food products is discovered in tumours of ALL breast cancer patients, The Daily Mail [online] Available: http://www.dailymail.co.uk/health/article-2085585/parabens-chemical-everyday-items-needs-investigationscientists-discover-tumours-all-breast-cancer-patients.html [09.10.13] 3. Scientific Committee on Consumer Safety (SCCS) (2011) Clarification on Opinion SCCS/1348/10 in the light of the Danish clause of safeguard banning the use of parabens in cosmetic products intended for children under three years of age, SCCS/1446/11, European Commissions 4. CosmeticOBS (2012) Phenoxyethanol: the French ANSM questions its safety for young children, [online] Available: http://www.cosmeticobs.com/news/cosmetic-news/phenoxyethanol-the-french-ansm-questions-its-safety-for-young-children--1217.html [09.10.13] 5. Cosmetics, Toiletry and Perfumery Association (CTPA) (2013) CTPA attends the British Association of Dermatologists (BAD) conference discussing the preservative MI, [online] Available: http://www.thefactsabout.co.uk/news.asp?newsid=244&view=yes&pageid=99&menu=sub&menuname=news&menuid=36&submenuid=9 9 [09.10.13] 6. Cosmetics Europe (2013) Cosmetics Europe Recommendation on use of MIT [online] Available: https://cosmeticseurope.eu/news-aevents/news.html [18.03.2014]

In The Personal Care Industry Thank You