Active.Lite Hair for Bleaching Efficacy Data

Transcription

1 Active.Lite Hair for Bleaching Efficacy Data Code: INCI Name: Polyquaternium-80 & Water & Pisum Sativum (Pea) Peptide & Selaginella Lepidophylla Extract CAS #: N/A & & & EINECS #: N/A & & & Cellular Viability Name of Study Results Active.Lite Hair was tested to evaluate its effects on the viability of normal human dermal fibroblasts (NDHF). At concentrations of 0.1% and 0.01% Active.Lite Hair, nor the preservatives contained therein, exhibited any inhibition of cell viability. It can be concluded that at normal use concentrations Active.Lite Hair is not cytotoxic. FT-IR Covalent Bond Formation Analysis Active.Lite Hair did not produce covalent bonds with L-cysteine or with L-cysteine in combination with hydrogen peroxide. The absence of S-C and S-N peaks and lack of decrease in S-H peaks indicate the lack of covalent bonding. It can therefore be concluded that at normal use concentrations, Active.Lite Hair will not produce covalent bonds with the free S-H groups from cysteine present in human hair. Hair Swatch Study The swatch study conducted demonstrates a clear difference in hair that has been treated, protected, and sealed using Active.Lite Hair for Color. When added to a hair color conditioner, Active.Lite Hair enhances the color deposit while also providing a silky soft feel. Hirox 3D Imaging Active.Lite Hair treated fibers are more comparable to the untreated virgin fiber, showcasing its ability to protect the hair from harsh chemical process and reduce overall damage to the fiber. Whereas the bleached swatches, both 30V and 40V fluoresce brightly to signify the extent of damage. Version#2/ info@activeconceptsllc.com +1 (704) Fax: +1 (704)

2 Salon Half Head Study Scanning Electron Microscopy The results of the assessment indicate that when incorporated into a shampoo, 2.0% Active.Lite Hair did show improvement in wet combability. However, when used in a conditioner Active.Lite Hair is capable of improving smoothing, wet and dry combability, anti-frizz, overall feel, shine and hydration more than the control conditioner. Standard Electron Microscopy (SEM) imaging shows high resolution images of the hair cuticles on each hair swatch. When the untreated images are compared to both the Active.Lite Hair treated swatches, a significant decrease in damage of the cuticle is exhibited. The Active.Lite Hair treated swatches display a smooth cuticle protected by the matrix style scaffolding for the promotion of healthy cuticles, added protection, and visibly healthier hair. Tensile Strength Data Tenacity is the customary measure of strength of a fiber usually defined as the ultimate (breaking) force of the fiber (in gram-force units) divided by the denier. The results above indicate that hair treated with bleach + Active.Lite Hair elicited results similar to that of untreated hair with less than one point difference after treatment. Elongation at break, also known as fracture strain, is the ratio between changed length and initial length after breakage of the test specimen expressing the capability of a material to resist changes of shape without crack formation, how much a hair fiber will stretch before it breaks. Both the results for 40V and 30V treated hair with Active.Lite Hair exhibited and improved elongation at break compared to bleached hair alone. Version#2/ info@activeconceptsllc.com +1 (704) Fax: +1 (704)

3 Cellular Viability Assay Analysis Phone: Fax: Tradename: Active.Lite Hair Code: CAS #: N/A & & & Test Request Form #: 1459 Lot #: Sponsor: Active Concepts, LLC; 107 Technology Drive Lincolnton, NC Study Director: Erica Segura Principle Investigator: Meghan Darley Test Performed: Cellular Viability Assay Introduction The cellular viability assay is useful for quantitatively measuring cell-mediated cytotoxicity, cell proliferation and mitochondrial metabolic activity. Increased metabolism in a cell indicates ample cellular respiration and adenosine triphosphate (ATP) production. ATP is the molecular energy of cells and is required in basic cell function and signal transduction. A decrease in ATP levels indicates cytotoxicity and decreased cell function while an increase in ATP levels indicates healthy cells. The cellular viability assay was conducted to assess the ability of Active.Lite Hair to increase cellular metabolic activity in cultured dermal fibroblasts. Assay Principle The assay utilizes a nonfluorescent dye, resazurin, which is converted to a fluorescent dye, resorufin, in response to chemical reduction of growth medium from cell growth and by respiring mitochondria. Healthy cells that are in a proliferative state will be able to easily convert resazurin into resorufin without harming the cells. This method is a more sensitive assay than other commonly used mitochondrial reductase dyes such as MTT. An increase in the signal generated by resazurin-conversion is indicative of a proliferative cellular state. This information is presented in good faith but is not warranted as to accuracy of results. Also, freedom from patent infringement is not implied. This information is offered solely for your investigation, verification, and consideration. Page 1 of 3 Version#2/ /Form#64

4 Cellular Viability Assay Analysis Phone: Fax: Materials A. Kit: PrestoBlue Cell Viability Reagent (Invitrogen, A13261) B. Incubation Conditions: 37 C at 5% CO 2 and 95% relative humidity (RH) C. Equipment: Forma humidified incubator; ESCO biosafety laminar flow hood; Light microscope; Pipettes D. Cell Line: Normal Human Dermal Fibroblasts (NHDF) (Lonza; CC-2511) E. Media/Buffers: Basal Medium (Fibrolife; LM-0001), 500µg/mL Human Serum Albumins (Fibrolife; LS-1001), 0.6µM Linoleic Acid (Fibrolife; LS-1001), 0.6µg/mL (Fibrolife; LS-1001), 5ng/mL Fibroblast Growth Factor (Fibrolife; LS- 1002), 5mg/mL Epidermal Growth Factor (Fibrolife; LS-1003), 30pg/mL Transforming Growth Factor β-1 (Fibrolife; LS-2003), 7.5mM L-Glutamine (Fibrolife; LS-1006), 1µg/mL Hydrocortisone Hemisuccinate (Fibrolife; LS-1007), 50µg/mL Ascorbic Acid (Fibrolife; LS-1005), 5µg/mL Insulin (Fibrolife; LS-1004) F. Culture Plate: Falcon flat bottom 96-well tissue culture treated plates G. Reagents: PrestoBlue reagent (10X) H. Other: Sterile disposable pipette tips Methods Human dermal fibroblasts were seeded into 96-well tissue culture plates and allowed to grow to confluency in complete serum-free media. A 10-fold serial dilution was performed resulting in Active.Lite Hair concentrations of 0.1% and 0.01% in complete serum-free media and incubated with fibroblasts for 24 hours. Ten microliters of viability reagent was added to 90µL of cell culture media in culture wells and a fluorometric measurement was taken at 560nm for excitation and 590nm for emission. This information is presented in good faith but is not warranted as to accuracy of results. Also, freedom from patent infringement is not implied. This information is offered solely for your investigation, verification, and consideration. Page 2 of 3 Version#2/ /Form#64

5 Cellular Viability Assay Analysis Phone: Fax: Results The data obtained from this study met criteria for a valid assay and the controls performed as anticipated. Active.Lite Hair did not exhibit negative effects on cell metabolism. Cellular metabolism results are shown as mean fluorescence units (MFU) and expressed as percentage change, calculated by the below equation: PPPPPPP (%)Chaaaa = MMM CCCCCCC MMM SSSSSS MMM CCCCCCC 100 Viability Assay Active.Lite Hair Relative Flouresence Units (RFU) % 0.01% Complete Media Active.Lite Hair RFU % Change 0.00% 0.52% -4.33% 100% 80% 60% 40% 20% 0% -20% -40% -60% -80% -100% Percent (%) Change Figure 1: Cellular Metabolism of Active.Lite Hair-treated fibroblasts expressed in terms of percent of control. Discussion In this study, Active.Lite Hair (code 22045) was tested to evaluate its effects on the viability of normal human dermal fibroblasts (NDHF). At concentrations of 0.1% and 0.01% Active.Lite Hair, nor the preservatives contained therein exhibited any inhibition of cell viability. It can therefore be concluded that at normal use concentrations Active.Lite Hair is not cytotoxic. This information is presented in good faith but is not warranted as to accuracy of results. Also, freedom from patent infringement is not implied. This information is offered solely for your investigation, verification, and consideration. Page 3 of 3 Version#2/ /Form#64

6 FT-IR Covalent Bond Formation Analysis Phone: Fax: Tradename: Active.Lite Hair Code: CAS #: N/A & & & Test Request Form #: 1538 Lot #: 43329P Sponsor: Active Concepts, LLC; 107 Technology Drive Lincolnton, NC Study Director: Erica Segura Principle Investigator: Meghan Darley Test Performed: FTIR Covalent Bond Formation Introduction Sulfur atoms typically join together to form strong covalent bonds, creating a link between two proteins. These disulfide bonds are partially responsible for the overall shape and strength of hair. Hair keratin proteins are made of thiol-bearing amino acids such as cysteine. Thiols are a class of compounds that contain sulfhydryl functional groups, composed of a sulfur atom and a hydrogen atom. Hair protein cleavage can be demonstrated by an increase in those free sulfhydryl groups (S-H) and general loss of other structural bonds (S-C or S-N). Chemical processing contributes to the weakening of hair over time. Relaxing or bleaching the hair fiber works by breaking the structural disulfide bonds present in hair proteins. The bends in curly hair are a product of a similar amino acid, cystine. Cystine units with disulfide bonds are formed when thiol groups of two cysteine residues are brought near each other in the course of protein folding, via oxidation reactions. Relaxers and bleaching techniques take advantage of the readily oxidizing capabilities of these cysteine residues. For example, the chemicals used in hair relaxing processes work by reducing cystine disulfide bridges to free cysteine S-H groups. Alternately, chemicals that curl hair work by oxidizing cysteine S-H groups to form cystine disulfide bonds. Another intermediate in thiol-bearing cysteine metabolism is cysteic acid. Larger amounts of cysteic acid have been reported in hydrolysates of tip ends of human hair, than in root ends due to oxidative stress and damage as measured via infrared spectroscopy. Continued use of powerful processing reagents such as hydrogen peroxide will in turn yield sulfonic acids. Analyzing the structure and bond strength of hair protein hydrolysates can thus provide models of hair damage and mechanisms by which treatments may work or bind to the hair. This information is presented in good faith but is not warranted as to accuracy of results. Also, freedom from patent infringement is not implied. This information is offered solely for your investigation, verification, and consideration. Page 1 of 3 Version#2/

7 FT-IR Covalent Bond Formation Analysis Phone: Fax: Molecular structure and bonding are modeled via infrared absorption spectroscopy using a Fourier Transform Infrared Spectrometer (FT-IR). The FT-IR method is used to determine the structure of molecules by measuring the vibrational energy as indicated by characteristic and selective absorption of IR radiation at specific wavelengths. FT-IR is useful for measuring the bond-strength of a specific treatment to damaged hair because it is a direct indication of bond intensity and formation. When a covalent bond is formed, shrinkage in S-H peak absorption ( cm-1) and re-appearance of S-C ( cm-1) or S-N ( cm-1) peaks are detected. If these patterns are not observed via FT-IR, then that is a direct indication the test material did not produce covalent bonds with the free S-H groups present in human hair. o Robbins, C. R. (2002). Chemical and physical behavior of human hair (4th ed.). New York, NY: Springer o Manuszak, M. A., et al (1996). The kinetics of disulfide bond reduction in hair by ammonium thioglycolate and dithiodiglycolic acid. Journal of the Society of Cosmetic Chemists, 47: o Kim, M. M. (2011). Effect of procyandin oligomers on oxidative hair damage. Skin Research and Technology, 17: o Jachowics, J. (1987). Hair damage and attempts to its repair. Journal of the Society of Cosmetic Chemists, 38: o Guarrera, L. et al. (1999). Ligand-linked changes at the subunit interfaces in Scapharca hemoglobins probed through the sulfhydryl infrared absorption. Biochemistry, 38(31): o Thermo Fisher Scientific. (2009). Infrared Spectral Interpretation Knowledge Base Technical Support Pamphet. o Oae, S. (1977). Organic Chemistry of Sulfur (1st ed.). New York, NY: Plenum Press Assay Principle The objective of this study was to demonstrate that Active.Lite (code 22045) does not produce covalent bonds with the free S-H groups from thiol-bearing cysteine in human hair, via FT-IR Spectroscopy. After application of Active.Lite using aqueous mixtures of cysteine and Active.Lite compared to a solution of Active.Lite, cysteine, and bleach (hydrogen peroxide), the absence of S-C and S-N peaks and lack of decrease in S-H peaks indicates that covalent bonds do not form with the free S-H groups in human hair. Materials A. Equipment: Nicolet is5 FT-IR Spectrophotometer; id5 ATR Accessory B. Reagents: Deionized Water; L-Cysteine; Hydrogen Peroxide C. Other: Disposable pipettes Methods Four solutions were made as follows: 1. Deionized Water 2. 10% Active.Lite Hair in Deionized Water 3. 10% Active.Lite Hair in Deionized Water + 1% L-Cysteine 4. 10% Active.Lite Hair in Deionized Water + 1% L-Cysteine + 3% Hydrogen Peroxide The samples were analyzed individually on the Nicolet is5 FT-IR with the id5 ATR laminated crystal accessory. Scans were analyzed for a decrease in S-H peaks and the appearance of S-C or S-N peaks. This information is presented in good faith but is not warranted as to accuracy of results. Also, freedom from patent infringement is not implied. This information is offered solely for your investigation, verification, and consideration. Page 2 of 3 Version#2/

8 FT-IR Covalent Bond Formation Analysis Phone: Fax: Results The data obtained from this study met criteria for a valid assay. Active.Lite Hair at a concentration of 10% did not produce covalent bonds with L-cysteine and hydrogen peroxide. 10% Active.Lite Hair in Deionized Water Deionized Water 10% Active.Lite Hair in Deionized Water + 1% L-Cysteine 10% Active.Lite Hair in Deionized Water + 1% L-Cysteine + 3% Hydrogen Peroxide Figure 1: FTIR Scan. 10% Active.Lite Hair in Deionized Water (Red), Deionized Water (Green), 10% Active.Lite Hair in Deionized Water + 1% L-Cysteine (Blue), 10% Active.Lite Hair in Deionized Water + 1% L-Cysteine + 3% Hydrogen Peroxide (Orange). Discussion As shown in figure 1, Active.Lite Hair (code 22045) did not produce covalent bonds with L-cysteine or with L-cysteine in combination with hydrogen peroxide. The absence of S-C and S-N peaks and lack of decrease in S-H peaks indicate the lack of covalent bonding. It can therefore be concluded that at normal use concentrations Active.Lit e Hair will not produce covalent bonds with the free S-H groups from cysteine present in human hair. This information is presented in good faith but is not warranted as to accuracy of results. Also, freedom from patent infringement is not implied. This information is offered solely for your investigation, verification, and consideration. Page 3 of 3 Version#2/

9 Active.Lite Hair for Bleaching Hair Swatch Study Patent Pending: Application Number 62/289,493 Code Number: Powerful Protection Composite Particles Natural Hair Scaffolding Strengthening Selective Matrix ABSTRACT Let us start from the beginning. The hair contains keratin proteins made up of the amino acid cysteine, which contains a sulfur atom. Typically sulfur atoms join together to form a disulfide bond to create a link between two proteins. In terms of the hair, proteins are partially responsible for your hair s overall shape and strength. When we perm or straighten our hair, these disulfide bonds are broken, thus affecting the strength and appearance of the hair. However, disulfide bonds can also break from repeated stress, such as exposure to heat, sunlight, hair styling and chemical treatments like coloring or perming. The question is how, do we prevent the hair from being damaged during times of stress? Active Concepts, LLC has developed a unique product comprised of two of our most popular materials Phytofuse Renew and ACB Pisum Sativum Peptide. Phytofuse Renew is a product derived from the desert plant Selaginella lepidophylla, often referred to as the rose of Jericho. It contains unique secondary metabolites that are required to endure its harsh desert environment. We isolate the plant s secondary metabolite system to get extraordinary benefits. Its smooth feel makes a perfect natural silicone replacement and it offers benefits, such as moisturization, repair, antioxidant properties, smoothness and shine when used in hair care products. We combine this material with ACB Pisum Sativum Peptide, which is an innovative, hydrolyzed pea protein. Great for any application but particularly unique when it comes to benefits to the hair. This anti-aging material can improve shine, combability, softness and manageability of the hair. Its exclusive manufacturing process contributes to its unique benefits - antioxidant activity and a volumizing effect to the hair! These attributes are not typical of a hydrolyzed protein, but are much needed in hair care. INCI Name: Polyquaternium-80 & Water & Hydrolyzed Pea Protein & Selaginella Lepidophylla Extract INCI Status: Approved REACH Status: Complies CAS Number: N/A & & & EINCS Number: N/A & & N/A & Origin: Botanical Processing: GMO Free No Ethoxylation No Irradiation No Sulphonation Additives: Preservatives: None Antioxidants: None Other additives: None Solvents Used: Water Appearance: Clear to Slightly Hazy Semi-Viscous Liquid Soluble/ Miscible: Water Soluble 100% Biodegradability Microbial Count: < 100 opg, No Pathogens Suggested Use Levels: Reference Formulation Guidelines on Page 5 Suggested Applications: Hair Protection, Nourishing, Support Benefits of Active.Lite Hair Protects Hair Great for All Hair Types Provides Moisturization Strengthens Hair As mentioned, proteins contain amino acids, such as cysteine. We increase the bioavailability of ACB Pisum Sativum Peptide with our manufacturing process and Version#3/ page 1/5

10 Active.Lite Hair for Bleaching Hair Swatch Study Patent Pending: Application Number 62/289,493 cysteine within the product. When added to a hair bleach product, Active.Lite Hair protects the hair from the damaging effects of bleach while also providing a silky soft feel. Achieve the hair color you have always wanted with Active.Lite Hair. MATERIALS AND METHODS This study was conducted by salon professionals using virgin hair swatches. The study shows there is no difference in processing time between the virgin bleached hair and the virgin bleached hair with Active.Lite Hair using 20 volume of developer. One set of hair swatches only used Pravana bleach (no additive) while another set used Pravana bleach combined with Active.Lite Hair. Virgin bleached hair and Active.Lite Hair had the same processing time of 1 hour and 47 minutes to reach the same color shade. Both hair swatches were rinsed with warm water and air dried. All measurements are with DRY Pravana bleach powders. Active.Lite Hair is not to be mixed with DRY powder bleach, however, the developer must be added and mixed to a butter cream consistency before adding Active.Lite Hair to lightener. Active.Lite Hair will not dilute the peroxide and it is not necessary to increase peroxide levels to achieve maximum hair lightening. Please see the last page for standard formulation guidelines. Standard Electron Microscopy (SEM) imaging was used to show high resolution images of the hair cuticles on each hair swatch. SEM images show how extensive hair damage is to the cuticle when bleach is applied. RESULTS Active.Lite Hair gives you the ability to chemically alter, thermally style, or enhance daily hair mechanics while still maintaining the hair s integrity by penetrating and strengthening internally. Active.Lite Hair exhibits both visual and sensorial properties that are most commonly associated with healthy, vibrant hair, specifically when compared to untreated (bleach only) controls. This swatch study conducted by salon professionals demonstrates a clear difference in hair that has been treated, protected, and sealed using Active.Lite Hair. Active.Lite Hair should be added to a bleach formulation to protect the hair from breakage during the bleaching/coloring process. Achieve flawless hair! Version#3/ page 2/5

11 Active.Lite Hair for Bleaching Hair Swatch Study Patent Pending: Application Number 62/289,493 Original Hair Color Figure 1. Original Hair Color. Bleached Hair (no additive) Bleached Hair with Active.Lite Hair 40 volume Figure 2. Bleached Hair with no additive. 40 volume Figure 3. Bleached Hair with Active.Lite Hair. Version#3/ page 3/5

12 Active.Lite Hair for Bleaching Hair Swatch Study Patent Pending: Application Number 62/289,493 Figure 4. SEM Imagery, Untreated Hair (Left) vs. Bleached Hair (Right) As seen in Figure 4, the Standard Electron Microscopy imaging at 20 kv demonstrates that, untreated hair is already prone to damage from everyday stressors, showing characteristic signs of breakdown. Results of bleaching the hair can be viewed on the right. The imagery results show more extensive damage to the hair fiber. This type of damage leads to irregular growth, breakages, and overall unhealthy or dead appearance. At a singular level, one cuticle may not seem important, but these strand to strand imperfections contribute to a much bigger picture of unhealthy and unprotected hair. Figure 5. SEM Imagery, cuticle treated with Active.Lite Hair Treating hair with Active.Lite Hair, as shown in Figure 5, makes a difference at the microscopic level, creating a scaffolding-like matrix to support hair while allowing the permeation of small molecules for hair bleaching. In Figure 4 the cuticle is clearly damaged whereas, in Figure 5 Active.Lite Hair demonstrates its ability to protect each fiber by shielding the cuticle, leaving it beautifully smoothed and annealed, contributing to overall healthier looking, and feeling hair. Version#3/ page 4/5

13 Active.Lite Hair for Bleaching Formulating with Active.Lite Hair Active.Lite Hair is an incredible, scientific breakthrough that is slated to shake the very foundation on which we ve built preventative hair care. Below, various guidelines will help you achieve the best formulation/use results: 1) Measure 2.5% Active.Lite Hair compared to DRY bleach powder Example: 1/2oz or less DRY bleach = 0.378g of Active.Lite Hair Example: 1oz or more DRY bleach = 0.758g of Active.Lite Hair 2) Measure approximately 25.0% water compared to dry bleach powder Example 1.0oz dry bleach (28.0 grams) = 6.4g of water Add 3.2g of water for 1/2oz or less of DRY bleach powder Add 6.4g of water for 1.0oz or more of DRY bleach powder 3) Combine Active.Lite Hair from Step 1 and water measuired from Step 2 4) Add developer to bleach powder to form desired consistency, If no benchmark exists a butter cream or gravy can be used as a target 5) Add the measured Active.Lite Hair from Step 3 with bleach from step 4 Active.Lite Hair can also be used in conditioners and sealers! Conditioners and sealers can be incorporated anywhere from % during the water phase of formulating. THINGS TO NOTE: This formulating guideline is strictly for Bleach Powder Lighteners. Do NOT add Active.Lite Hair to the dry bleach powder. Follow aforementioned guidelines for optimal results. Active Concepts, LLC Lincolnton, NC. USA www. activeconceptsllc.com Office: +1 (704) info@activeconceptsllc.com Active Concepts S.r.l. Milano ITALY Tel info@activeconcepts.it Active Concepts LLC, Asia Kaohsiung, Taiwan Tel josephyeh@activeconceptsllc.com Information contained in this technical literature is believed to be accurate and is offered in good faith for the benefit of the customer. The company, however, cannot assume any liability or risk involved in the use of its chemical products since the conditions of use are beyond our control. Statements concerning the possible use of our products are not intended as recommendations to use our products in the infringement of any patent. We make no warranty of any kind, expressed or implied, other than that the material conforms to the applicable standard specification. Freedom from patent infringement is not implied. All information is for investigative purposes only. page 5/5

14 Hirox 3D Imaging +1 (704) Fax: +1 (704) Tradename: Active.Lite Hair Code: Test Request Form #: 1404 Test Performed: Hirox 3D Imaging Methods & Materials This study was conducted by salon professionals using Sensationnel Bare & Natural Brazilian 100% Virgin Remi Unprocessed Human Hair (Hair Zone Moonachie, NJ). One swatch, left unaltered, was analyzed as the control. Two test swatches were bleached alone with Pravana bleach, one with 30V and the other with 40V. Once the bleach processed, the swatches were washed and blown dry. The two treated swatches were treated as follows; Bleach (30V) + 1/4oz. 10% solution of Active.Lite Hair and Bleach (40V) + 1/4oz 10% solution of Active Lite Hair. Once the bleach processed, the swatches were washed and blown dry. Note: Active.Lite Hair was not mixed with the dry powder bleach alone, the developer must be added and mixed to a butter cream consistency before adding Active.Lite Hair. Manufacturing Solutions Center (MSC) located in Conover, North Carolina was asked to perform Hirox 3D Imaging on the five hair swatches provided by Active Concepts, LLC. MSC utilized a KH-7700 Hirox 3D Imaging Microscope to perform the test. The lens used was MX(G)-5040Z with magnification ranging from 50x-300x. This method was used in order to visually see thermal damage induced by a chemical hair relaxer treatment vs. chemical hair relaxer treatment + Active.Lite Relaxer. The sample identifications are listed below. Parameter Test Sample ID Style Untreated Virgin T UV Hair + Bleach (40V) T V Hair + Bleach (30V) T V Hair + Bleach (40V) + 10% Active.Lite Hair T VA Hair + Bleach (30V) + 10% Active.Lite Hair T VA Information contained in this technical literature is believed to be accurate and is offered in good faith for the benefit of the customer. The company, however, cannot assume any liability or risk involved 1in the use of its chemical products since the conditions of use are beyond our control. Statements concerning the possible use of our products are not intended as recommendations to use our products in the infringement of any patent. We make no warranty of any kind, expressed or implied, other than that the material conforms to the applicable standard specification. Page 1 of 27 Version#1/ /Form#90

15 Manufacturing Solutions Center 301 Conover Station SE Conover, NC Maureen Danaher Active Concepts, LLC 107 Technology Drive Lincolnton, NC RECEIVED DATE: August 12, 2015 COMPLETED DATE: August 17, 2015 REFERENCE: TS Sample Type: Hair Style: T UV Sample Form: Hair Swatches Color: Black Size: N/A Description: Hair Swatch for picture top, middle and end of strand, to check for heat damage. TEST RESULTS: Hirox Microscopic Examination: MSC Test Method for Microscopic Examination of Samples on the Hirox 3D Imaging Digital Microscope ( ) Top of swatch Comments: A picture was taken at top, middle and end of each hair swatch. Then two hairs choosen at random from each area was photograhed, at top, middle and end. TS (Hirox examination, T UV, Blk Hair) Page 1 of 6

16 Manufacturing Solutions Center 301 Conover Station SE Conover, NC st Hair top 2nd Hair top TS (Hirox examination, T UV, Blk Hair) Page 2 of 6

17 Manufacturing Solutions Center 301 Conover Station SE Conover, NC Middle of swatch 1st Hair middle TS (Hirox examination, T UV, Blk Hair) Page 3 of 6

18 Manufacturing Solutions Center 301 Conover Station SE Conover, NC nd Hair, middle End of swatch TS (Hirox examination, T UV, Blk Hair) Page 4 of 6

19 Manufacturing Solutions Center 301 Conover Station SE Conover, NC st Hair end 2nd Hair end Testing Information: Samples taken approximately as dictated by defect or customer Examine is defined by defect, sample or customer TS (Hirox examination, T UV, Blk Hair) Page 5 of 6

20 Manufacturing Solutions Center 301 Conover Station SE Conover, NC Maureen Danaher Active Concepts, LLC 107 Technology Drive Lincolnton, NC RECEIVED DATE: August 12, 2015 COMPLETED DATE: August 17, 2015 REFERENCE: TS Sample Type: Hair Style: T V Sample Form: Hair Swatches Color: Blonde Size: N/A Description: Hair Swatch for picture top, middle and end of strand, to check for heat damage. TEST RESULTS: Hirox Microscopic Examination: MSC Test Method for Microscopic Examination of Samples on the Hirox 3D Imaging Digital Microscope ( ) Top of swatch Comments: A picture was taken at top, middle and end of each hair swatch. Then two hairs choosen at random from each area was photograhed, at top, middle and end. TS (Hirox examination, T V, Blonde Hair) Page 1 of 6

21 Manufacturing Solutions Center 301 Conover Station SE Conover, NC st Hair top 2nd Hair top TS (Hirox examination, T V, Blonde Hair) Page 2 of 6

22 Manufacturing Solutions Center 301 Conover Station SE Conover, NC Middle of swatch 1st Hair middle TS (Hirox examination, T V, Blonde Hair) Page 3 of 6

23 Manufacturing Solutions Center 301 Conover Station SE Conover, NC nd Hair, middle End of swatch TS (Hirox examination, T V, Blonde Hair) Page 4 of 6

24 Manufacturing Solutions Center 301 Conover Station SE Conover, NC st Hair end 2nd Hair end Testing Information: Samples taken approximately as dictated by defect or customer Examine is defined by defect, sample or customer TS (Hirox examination, T V, Blonde Hair) Page 5 of 6

25 Manufacturing Solutions Center 301 Conover Station SE Conover, NC Maureen Danaher Active Concepts, LLC 107 Technology Drive Lincolnton, NC RECEIVED DATE: August 12, 2015 COMPLETED DATE: August 17, 2015 REFERENCE: TS Sample Type: Hair Style: T V Sample Form: Hair Swatches Color: Blonde Size: N/A Description: Hair Swatch for picture top, middle and end of strand, to check for heat damage. TEST RESULTS: Hirox Microscopic Examination: MSC Test Method for Microscopic Examination of Samples on the Hirox 3D Imaging Digital Microscope ( ) Top of swatch Comments: A picture was taken at top, middle and end of each hair swatch. Then two hairs choosen at random from each area was photograhed, at top, middle and end. TS (Hirox examination, T V, Blonde Hair) Page 1 of 6

26 Manufacturing Solutions Center 301 Conover Station SE Conover, NC st Hair top 2nd Hair top TS (Hirox examination, T V, Blonde Hair) Page 2 of 6

27 Manufacturing Solutions Center 301 Conover Station SE Conover, NC Middle of swatch 1st Hair middle TS (Hirox examination, T V, Blonde Hair) Page 3 of 6

28 Manufacturing Solutions Center 301 Conover Station SE Conover, NC nd Hair, middle End of swatch TS (Hirox examination, T V, Blonde Hair) Page 4 of 6

29 Manufacturing Solutions Center 301 Conover Station SE Conover, NC st Hair end 2nd Hair end Testing Information: Samples taken approximately as dictated by defect or customer Examine is defined by defect, sample or customer TS (Hirox examination, T V, Blonde Hair) Page 5 of 6

30 Manufacturing Solutions Center 301 Conover Station SE Conover, NC Maureen Danaher Active Concepts, LLC 107 Technology Drive Lincolnton, NC RECEIVED DATE: August 12, 2015 COMPLETED DATE: August 17, 2015 REFERENCE: TS Sample Type: Hair Style: T VA Sample Form: Hair Swatches Color: Dk Blonde Size: N/A Description: Hair Swatch for picture top, middle and end of strand, to check for heat damage. TEST RESULTS: Hirox Microscopic Examination: MSC Test Method for Microscopic Examination of Samples on the Hirox 3D Imaging Digital Microscope ( ) Top of swatch Comments: A picture was taken at top, middle and end of each hair swatch. Then two hairs choosen at random from each area was photograhed, at top, middle and end. TS (Hirox examination, T VA, Dk Blonde Hair) Page 1 of 6

31 Manufacturing Solutions Center 301 Conover Station SE Conover, NC st Hair top 2nd Hair top TS (Hirox examination, T VA, Dk Blonde Hair) Page 2 of 6

32 Manufacturing Solutions Center 301 Conover Station SE Conover, NC Middle of swatch 1st Hair middle TS (Hirox examination, T VA, Dk Blonde Hair) Page 3 of 6

33 Manufacturing Solutions Center 301 Conover Station SE Conover, NC nd Hair, middle End of swatch TS (Hirox examination, T VA, Dk Blonde Hair) Page 4 of 6

34 Manufacturing Solutions Center 301 Conover Station SE Conover, NC st Hair end 2nd Hair end Testing Information: Samples taken approximately as dictated by defect or customer Examine is defined by defect, sample or customer TS (Hirox examination, T VA, Dk Blonde Hair) Page 5 of 6

35 Manufacturing Solutions Center 301 Conover Station SE Conover, NC Maureen Danaher Active Concepts, LLC 107 Technology Drive Lincolnton, NC RECEIVED DATE: August 12, 2015 COMPLETED DATE: August 17, 2015 REFERENCE: TS Sample Type: Hair Style: T VA Sample Form: Hair Swatches Color: Dk Blonde Size: N/A Description: Hair Swatch for picture top, middle and end of strand, to check for heat damage. TEST RESULTS: Hirox Microscopic Examination: MSC Test Method for Microscopic Examination of Samples on the Hirox 3D Imaging Digital Microscope ( ) Top of swatch Comments: A picture was taken at top, middle and end of each hair swatch. Then two hairs choosen at random from each area was photograhed, at top, middle and end. TS (Hirox examination, T VA, Dk Blonde Hair) Page 1 of 6

36 Manufacturing Solutions Center 301 Conover Station SE Conover, NC st Hair top 2nd Hair top TS (Hirox examination, T VA, Dk Blonde Hair) Page 2 of 6

37 Manufacturing Solutions Center 301 Conover Station SE Conover, NC Middle of swatch 1st Hair middle TS (Hirox examination, T VA, Dk Blonde Hair) Page 3 of 6

38 Manufacturing Solutions Center 301 Conover Station SE Conover, NC nd Hair, middle End of swatch TS (Hirox examination, T VA, Dk Blonde Hair) Page 4 of 6

39 Manufacturing Solutions Center 301 Conover Station SE Conover, NC st Hair end 2nd Hair end Testing Information: Samples taken approximately as dictated by defect or customer Examine is defined by defect, sample or customer TS (Hirox examination, T VA, Dk Blonde Hair) Page 5 of 6

40 Hirox 3D Imaging +1 (704) Fax: +1 (704) Discussion Hirox 3D Microscopic Examination is a test method for microscopic examination of hair samples. Damage of the hair fiber can be seen within these images in which the damaged areas of the fiber fluoresce. The more fluorescence a fiber exhibits, the more damaged. Within the images below significant less damage can be viewed on both the Untreated Virgin swatch and Active.Lite Hair treated swatches. Whereas the bleached swatches, both 30V and 40V fluoresce brightly signifying the extent of damage. The Active.Lite Hair treated fibers are more comparable to the untreated virgin fiber, showcasing Active.Lite Hair s ability to protect the hair fiber from the harsh chemical process and reducing overall damage to the fiber. Active.Lite Hair s ability to provide a selective bio-film allows bleach particles to enter the cortex while providing structure and protection leading to less damaged and improved overall post treatment fibers. Information contained in this technical literature is believed to be accurate and is offered in good faith for the benefit of the customer. The company, however, cannot assume any liability or risk involved 2in the use of its chemical products since the conditions of use are beyond our control. Statements concerning the possible use of our products are not intended as recommendations to use our products in the infringement of any patent. We make no warranty of any kind, expressed or implied, other than that the material conforms to the applicable standard specification. Page 2 of 27 Version#1/ /Form#90

41 Active.Lite Hair Salon Half-Head Hair Study Patent Pending: Application Number 62/289,493 Code Number: Powerful Protection Composite Particles natural Hair Scaffolding Strengthening Selective Matrix ABSTRACT The condition of the cuticle (the outer most layer of the hair) significantly affects both the manageability and sleekness of our hair. Overtime, hair can become damaged, which can result in the cuticle lifting because of both environmental and styling influences and processes. The result: lifeless, dull hair that is difficult to manage. Improving the sleekness of hair has been shown to instantly create a healthier more youthful appearance. Increasing combability not only eases manageability, but also helps to minimize physical damage that perpetuates the loss of body and difficulty in styling. Actve.Lite Hair is a product designed to be added to hair bleach for long lasting protection and chemical burn prevention. It gently lifts the hair and protects it from the harsh effects of bleach. This ingredient also enhances smoothing, wet and dry combability, anti-frizz, overall feel, shine and hydration of the hair. The purpose of this study was to confirm whether Active.Lite Hair is capable of providing benefits when included in a shampoo and conditioner on ethnic hair types. A half head study was conducted to determine the comparison of a control shampoo vs. 2.0% Active.Lite Hair in the control shampoo. Additionally, a comparison between the control conditioner and 2.0% Active.Lite Hair in the control conditioner were reported. Each volunteer s hair was photographed prior to the treatment and again after the shampoo and conditioner had been applied and the hair was styled. The images of the half head study were used in conjunction with a sensory assessment subjectively rating the parameters - cleansing, smoothing, dry and wet combability, anti-frizz, overall feel, shine and hydration. This assessment was conducted both before and after treatment. Based on the results obtained, Active.Lite Hair is capable of enhancing smoothing, wet and dry combability, antifrizz, overall feel, shine and hydration when used in a conditioner. These attributes makes it an ideal ingredient for use in products intended for thick, unruly or ethnic hair types. INCI Name: Polyquaternium-80 & Water & Hydrolyzed Pea Protein & Selaginella Lepidophylla Extract INCI Status: Approved REACH Status: Complies CAS Number: N/A & & & EINCS Number: N/A & & N/A & TRF#: S37 Lot Number(s): NC150929E & NC150029F Suggested Use Levels: Reference Formulation Guidelines on TDS Sponsor: Active Concepts, LLC 107 Technology Drive Lincolnton, North Carolina Study Director: Erica Segura Principle Investigator: Meghan Darley Suggested Applications: Hair Protectant, Nourishing, Supportditioning, Hydrating Benefits of Active.Lite Hair: Protects Hair Great for All Hair Types Provides Moisturization Strenthens Hair Version#3/ page 1/5

42 Active.Lite Hair Salon Half-Head Hair Study Patent Pending: Application Number 62/289,493 MATERIALS AND METHODS The study was conducted using five participants. Each subject had their baseline photo taken prior to having their hair washed. The participant was also asked to complete a survey rating their hair prior to treatment on a scale of 1 to 10, with 1 being the lowest and 10 being the highest, using the following parameters cleansing, smoothing, dry and wet combability, anti-frizz, overall feel, shine and hydration. Half of the head was treated with the control shampoo and conditioner while the other half of the head was treated with 2.0% Active.Lite Hair in the base shampoo and base conditioner. After the application and rinse of the test and positive control products, each participant s hair was blown dry using a round brush on both sides of the head. Once the hair was completely dry, the participant was asked to again assess the same parameters of both halves of their hair. Assessments were made using a rubric from 1 to 10, with 1 being the lowest and 10 being the highest. RESULTS Parameters Tested Assessment of the Control Shampoo Assessment of the Experimental (2.0% Active.Lite Hair in Control Shampoo) Assessment of the Control Conditioner Assessment of the Experimental (2.0% Active.Lite Hair in Control Conditioner) Cleansing X X Smoothing Wet Combability Dry Combability X X Anti-Frizz X X Overall Feel X X Shine X X Hydration X X Mean Chart 1. Average Results for Participant s Sensory Assessment Parameters Tested Percent Difference Comparison of Control Shampoo vs. Experimental (2.0% Active.Lite Hair in Control Shampoo) Percent Difference Comparison of Control Conditioner vs. Experimental (2.0% Active.Lite Hair in Control Conditioner) Cleansing 0% X Smoothing 0% 46% Wet Combability 18% 18% Dry Combability X 13% Anti-Frizz X 13% Overall Feel X 40% Shine X 40% Hydration X 29% Mean 18% 27% Chart 2. Percent Difference of Participant s Sensory Assessment Version#3/ page 2/5

43 Rating from 1 (worst) to 10 (best) Active.Lite Hair Salon Half-Head Hair Study Patent Pending: Application Number 62/289,493 Assessment of Hair Characteristics Cleansing Smoothing Wet Dry Combability Combability Anti-Frizz Overall Feel Shine Hydration Assessment of the Control Shampoo Assessment of the Experimental (2.0% Active.Lite Hair in Control Shampoo) Assessment of the Control Conditioner Assessment of the Experimental (2.0% Active.Lite Hair in Control Conditioner) Graph 1. Rating of hair characteristics following sensory assessment Half Head Treated with Control Shampoo and Conditioner Half-Head Treated with 2.0% Active.Lite Hair in a Base Shampoo & Conditioner Figure 1. Full head Baseline, Untreated Hair Figure 2. Half Head Treated Half Head Treated with Control Shampoo and Conditioner Half-Head Treated with 2.0% Active.Lite Hair in a Base Shampoo & Conditioner Figure 3. Full head Baseline, Untreated Hair Figure 4. Half Head Treated Version#3/ page 3/5

44 Active.Lite Hair Salon Half-Head Hair Study Patent Pending: Application Number 62/289,493 Half Head Treated with Control Shampoo and Conditioner Half-Head Treated with 2.0% Active.Lite Hair in a Base Shampoo & Conditioner Figure 5. Full head Baseline, Untreated Hair Figure 6. Half Head Treated Half Head Treated with Control Shampoo and Conditioner Half-Head Treated with 2.0% Active.Lite Hair in a Base Shampoo & Conditioner Figure 7. Full head Baseline, Untreated Hair Figure 8. Half Head Treated Half Head Treated with Control Shampoo and Conditioner Half-Head Treated with 2.0% Active.Lite Hair in a Base Shampoo & Conditioner Figure 9. Full head Baseline, Untreated Hair Figure 10. Half Head Treated Version#3/ page 4/5

45 Active.Lite Hair Salon Half-Head Hair Study Patent Pending: Application Number 62/289,493 When comparing hair characteristics of the baseline assessments to the post style assessments, the benefits of including 2.0% Active.Lite Hair in a conditioner are even more apparent. In relation to the baseline readings, the test-half of the head improved the intended subjective parameters of smoothness, wet and dry combability, anti-frizz, overall feel, shine and hydration by 46%, 18%, 13%, 13%, 40%, 40% and 29%, respectively. It is clear from the images in this study that Active.Lite Hair helps create a smooth, sleek hairstyle. Additionally, in all images, the hair is noticeably shinier, with less frizz and has a healthy appearance. The professional stylist who performed the actual tests by applying the product, styling the hair and documenting the images said Active.Lite Hair is great for use on any hair type. This product works well at reducing frizziness and smoothing of the hair. This product enhances the sleekness, combability, the overall feel and look of the hair. Recommended for conusmers with ethnic or coarse hair to achieve a silky, less frizzy style. Active.Lite Hair is good for use in a pre-treatment or after chemical treatment product. This product also has the ability to recondition the hair after bleaching, providng a smooth and healthy feel. Comparison of Control Conditioner vs. Experimental Hydration Wet Combability 40% 35% 30% 25% 20% 15% 10% 5% 0% Dry Combability Shine Anti-Frizz Overall Feel Graph 2. Hair Assessment results for sensory characteristics DISCUSSION The results of the assessment indicate that when incorporated into a shampoo, 2.0% Active.Lite Hair did show improvement in wet combability. However, when used in a conditioner Active.Lite Hair is capable of improving smoothing, wet and dry combability, anti-frizz, overall feel, shine and hydration more than the control conditioner. These results can be further supported by figures 1 through 10, where clearly the half of the subject s head treated with 2.0% Active.Lite Hair appears sleek, smooth and less frizzy. Additionally, the subjects reported a significant increase in smoothness, combability and overall feel of the hair, especially after chemical treatment. Active Concepts, LLC Lincolnton, NC. USA www. activeconceptsllc.com Office: +1 (704) info@activeconceptsllc.com Active Concepts S.r.l. Milano ITALY Tel info@activeconcepts.it Active Concepts LLC, Asia Kaohsiung, Taiwan Tel josephyeh@activeconceptsllc.com Information contained in this technical literature is believed to be accurate and is offered in good faith for the benefit of the customer. The company, however, cannot assume any liability or risk involved in the use of its chemical products since the conditions of use are beyond our control. Statements concerning the possible use of our products are not intended as recommendations to use our products in the infringement of any patent. We make no warranty of any kind, expressed or implied, other than that the material conforms to the applicable standard specification. Freedom from patent infringement is not implied. All information is for investigative purposes only. page 5/5

46 Scanning Electron Microscopy +1 (704) Fax: +1 (704) Tradename: Active.Lite Hair Code: Test Request Form #: 1404 Test Performed: Scanning Electron Microscopy (SEM) Methods & Materials This study was conducted by salon professionals using Sensationnel Bare & Natural Brazilian 100% Virgin Remi Unprocessed Human Hair (Hair Zone Moonachie, NJ). One swatch, left unaltered, was analyzed as the control. Two test swatches were bleached alone with Pravana bleach, one with 30V and the other with 40V. Once the bleach processed, the swatches were washed and blown dry. The two treated swatches were treated as follows; Bleach (30V) + 1/4oz. 10% solution of Active.Lite Hair and Bleach (40V) + 1/4oz 10% solution of Active Lite Hair. Once the bleach processed, the swatches were washed and blown dry. Note: Active.Lite Hair was not mixed with the dry powder bleach alone, the developer must be added and mixed to a butter cream consistency before adding Active.Lite Hair. Gaston College Textile Technology Center located in Belmont, North Carolina was asked to perform Scanning Electron Microscopy Imaging (SEM) on the five hair swatches provided by Active Concepts, LLC. Gaston College used a Zeiss DSM 962 to perform the test at 20.0kV using a magnification range from 50x-300x. This method utilizes an electron microscope that produces images a chemically treated hair by scanning the hair with a focused beam of electrons. These electrons interact with the atoms of the hair sample to provide images of the hairs surface topography and surface composition. The sample identifications are listed below. Parameter Test Sample ID Untreated Virgin Hair + Bleach (40V) Hair + Bleach (30V) Hair + Bleach (40V) + 10% Active.Lite Hair Hair + Bleach (30V) + 10% Active.Lite Hair F UV F V F V F VA F VA Information contained in this technical literature is believed to be accurate and is offered in good faith for the benefit of the customer. The company, however, cannot assume any liability or risk involved in the use of its chemical products since the conditions of use are beyond our control. Statements concerning the possible use of our products are not intended as recommendations to use our products in the infringement of any patent. We make no warranty of any kind, expressed or implied, other than that the material conforms to the applicable standard specification. 1 Page 1 of 3 Version#1/ /Form#71

47 Scanning Electron Microscopy +1 (704) Fax: +1 (704) Results Figure 1. Untreated Virgin-F UV Figure 2. Hair + Bleach (40V)-F V Figure 3. Hair + Bleach (30V)-F V Figure 4. Hair + Bleach (40V) + 10% Active.Lite Hair-F VA Information contained in this technical literature is believed to be accurate and is offered in good faith for the benefit of the customer. The company, however, cannot assume any liability or risk involved in the use of its chemical products since the conditions of use are beyond our control. Statements concerning the possible use of our products are not intended as recommendations to use our products in the infringement of any patent. We make no warranty of any kind, expressed or implied, other than that the material conforms to the applicable standard specification. 2 Page 2 of 3 Version#1/ /Form#71

48 Scanning Electron Microscopy +1 (704) Fax: +1 (704) Figure 5. Hair + Bleach (30V) + 10% Active.Lite Hair-F VA Discussion Standard Electron Microscopy (SEM) imaging shows high resolution images of the hair cuticles on each hair swatch. SEM images depict how the outermost layer of the hair, the cuticle, is effected by bleach treatments. The SEM imaging demonstrates that, untreated virgin hair is clearly already prone to damage from everyday aggressors, showing characteristic signs of breakdown, pre chemical treatment. The imagery results of bleached hair alone (both 30V and 40V) depict an extensively damaged cuticle. This type of damage leads to irregular growth, breakages, and overall unhealthy or dead appearance. At a singular level, one cuticle may not seem important, but these strand to strand imperfections contribute to a much bigger picture of unhealthy and unprotected hair. When the untreated images are compared to both the Active.Lite Hair treated swatches, a significant decrease in damage of the cuticle is exhibited. The Active.Lite Hair treated swatches display a smooth cuticle protected by the matrix style scaffolding for the promotion of healthy cuticles, added protection, and visibly healthier hair. It is this scaffolding and its semi-permeable membrane that promotes the exhibition of properties such as moisturization, ph balance, barrier protection, and additionally, protection from hair weakening after exposure to bleach rendering Active.Lite Hair the ideal addition to any bleach treatment. Information contained in this technical literature is believed to be accurate and is offered in good faith for the benefit of the customer. The company, however, cannot assume any liability or risk involved in the use of its chemical products since the conditions of use are beyond our control. Statements concerning the possible use of our products are not intended as recommendations to use our products in the infringement of any patent. We make no warranty of any kind, expressed or implied, other than that the material conforms to the applicable standard specification. 3 Page 3 of 3 Version#1/ /Form#71