KERATEC IFP PE

DS-197R-1

July 31, 2007

KERATEC IFP PE
Protects against the aging effects of environmental pollutants

INCI Name:

Keratin (and) Hydrolyzed Keratin

Previous studies with KERATEC IFP PE have shown the anti-aging efficacy against the suns rays (refer to Datasheet DS-181). The strength of hair fibers was maintained and the hair surface was protected from UV-related aging with KERATEC IFP PE treatment. This was achieved by the preferential binding/degradation and anti-oxidant activity of KERATEC IFP PE. The present document details the effectiveness of KERATEC IFP PE against the attack of other environmental aggressors. The results of hair surface analysis and combing force experiments show that KERATEC IFP PE gives substantial protection from the damaging effects of the ozone, sulphur dioxide and smoke. KERATEC IFP PE is a purified protein fraction isolated from pure New Zealand wool. Specifically, it is a multi-component soluble form of Intermediate Filament Protein. The proprietary manufacturing process used to manufacture KERATEC IFP PE generates intact keratin proteins with a molecular weight range in the region of approximately 40,000 60,000 Daltons (determined by SDS polyacrylamide gel electrophoresis) and keratin peptides with molecular weight in the range 3000 4000 Daltons (determined by size exclusion HPLC). Intermediate filament protein was selected as a specific target for solubilization due to its role in natural hair fibers as the protein that gives hair its structural integrity. The range of protein molecular weights present in KERATEC IFP PE allows the formation of coherent films on the cuticle surface and penetration into the cortex.

Croda Inc 300-A Columbus Circle, Edison, NJ 08837 Tel 732-417-0800 Fax 732-417-0804 Website www.crodausa.com Sederma, Inc. Tel 732-692-1652 Fax 732-417-0804 Croda Inc West Coast Sales Office 9545 Santa Anita Ave, Rancho Cucamonga, CA 91730 Tel 909-9802510 Fax 909-476-7304 Croda Miami Inc 200 South Park Road, Suite 303, Hollywood, FL 33021 Tel 1-888-84CRODA Fax 954-989-8186 Sol Kaplan & Son PO Box 240234 Memphis TN 38124-0234 Tel 901-685-0323 Fax 901-763-3612 Croda Canada Ltd 221A Racco Parkway, Vaughan, Ontario L4J8X9 CANADA Tel 905-886-1383 Fax 905-886-4753 The information in this publication is believed to be accurate and is given in good faith, but no representation or warranty as to its completeness or accuracy is made. Suggestions for uses or applications are only opinions. Users are responsible for determining the suitability of these products for their own particular purpose. No representation or warranty, expressed or implied, is made with respect to information or products including, without limitation, warranties of merchantability, fitness for a particular purpose, non-infringement of any third party patent or other intellectual property rights including, without limit, copyright, trademark and designs. Any trademarks identified herein are trademarks of the Croda group of companies.

KERATEC IFP PE

Operating through a sacrificial degradation and antioxidant mechanism (by virtue of its high level of cystine derived amino acids), KERATEC IFP PE offers effective protection against the aging effects of common pollutants. It is able to do this because the reactions which damage the hair i.e. oxidation, bond cleavage, radical attack etc. take place at the protective KERATEC IFP PE surface layer (which is made of the same material) and therefore the underlying fiber remains undamaged. Hair is a complex system consisting principally of proteins and lipids. In youthful hair these lipids and proteins function to keep the hair healthy, vibrant and strong. Prolonged exposure to the destructive factors of the environment, such as sunlight and pollution, leads to attack and breakdown of the structural components of the hair. This breakdown is primarily through the cross-linking of the keratin proteins which subsequently causes brittleness, reduced lubricity and disruption of the cuticle. This damage dulls and weakens the hair and makes it more vulnerable to further attack. The air in our cities is being polluted by cars, trucks and through the burning of fossil fuels. Air pollution even comes from everyday activities such as dry cleaning and filling your car with petrol. Air pollution is a problem for all of us. Many air pollutants remain in the environment for long periods of time and are carried by the winds hundreds of miles from their origin. Primary pollutants produced by human activity include: oxides of sulphur, nitrogen and carbon organic compounds, such as hydrocarbons, which lead to ozone particulate matter, such as smoke and dust Benefits KERATEC IFP PE operates through a sacrificial and chemical protection process to combat the aggressive species found in environmental pollution and in doing so: Keeps hair surface healthy Maintains hair manageability and youthfulness Acts as a sacrificial protective layer on the hair

Protection from Environmental Stresses Sulphur dioxide Sulphur dioxide in the atmosphere arises from both natural and human activities. Natural processes include decomposition and combustion of organic matter. The main human activities are the burning of coal, high sulphur oil and diesel fuel. SO2 and the pollutants formed from SO2 can be transported over long distances and deposited far from the point of origin. This means that problems with SO2 are not confined to areas where it is emitted, and SO2 accounts for a significant component of the pollutant load in many cities.

KERATEC IFP PE

Ozone Near the ground, ozone is a colorless, gaseous secondary pollutant. Unlike other pollutants, ozone is not directly emitted from man-made sources in large quantities. Ozone occurs naturally in the atmosphere, but the chemical reaction between VOCs (volatile organic compounds) from vehicles and industry, nitrogen oxides and sunlight can produce enough ground level ozone to affect crops and people. Ozone is also produced during lightning storms and by photocopiers with maximum concentrations generally occurring downwind of source emissions. Ozone is a component of photochemical smog.

It is well known that ozone is an aggressive oxidizing agent which is very reactive towards biological materials attacking amino acids like cystine and tryptophan, which are commonly found in hair. This can result in cleavage of the bonds responsible for holding the hair fiber together.
The type of hair fiber damage caused by ozone can be seen in figure 1.

Figure 1: SEM of wet hair exposed to 0.1ppm ozone for 30s

Smoke Smoke is just one of the constituents of particulate matter in the air. The most obvious source of smoke is from cigarettes and it is known to attack biological tissues. Tobacco smoke contains more than 4000 substances including formaldehyde, lead, carbon monoxide, ammonia and benzene, many of which are toxic and highly reactive towards biological material. Accelerated aging in hair is a symptom of smoke exposure caused by oxidative and free radical damage.

KERATEC IFP PE

Ozone exposed hair Tryptophan Fluorescence

The degradation of tryptophan in the hair cuticle is a standard method of indicating the amount of damage to the surface of the hair and also for determining the ability of a protective material to prevent the hair fiber from environmental damage1. Fluorescence analysis of tryptophan emission was performed in duplicate on ozone exposed hair (De Meo Bros. medium brown). The following conditions were used: Untreated baseline (control) hair fibers Hair fibers treated with conditioner base - rinse off Hair fibers treated with conditioner base + KERATEC IFP PE (1% as supplied) - rinse off

The wet hair fibers were exposed to ozone at 20ppm for 2 minutes. The experiment involved card mounted fibers, a rinse off protocol and a post surfactant wash following ozone exposure and prior to measurement. The hair was dampened and cut into 2mm sections and 50 mg spread evenly over the face of a quartz solid state cell. Spectra were recorded using an excitation wavelength of 295nm (UVB) and the resultant emission spectra gathered across a wavelength range of 310 540nm. The results of the experiment are shown below (Figure 2).

Tryptophan Damage of Ozone Exposed Hair Fibers

Normalized Intensity of emitted light

100 95 90 85 80 75 70 Untreated control Conditioner base Conditioner base + 1% Keratec IFP 1 % Keratec IFP PE
Increasing surface damage 4

Figure 2: Graph showing tryptophan degradation caused by exposure to ozone; 1% KERATEC IFP PE is able to protect the hair fiber

KERATEC IFP PE

The graph clearly shows that exposure to ozone significantly reduces the emission intensity of the untreated control hair indicating major degradation of tryptophan levels, giving rise to compromised cuticle integrity. The conditioner base protected the hair to a certain extent, but KERATEC IFP PE provides 100% protection and there is no degradation of hair fiber tryptophan. This demonstrates that the high molecular weight component of KERATEC IFP PE is providing a protective film on the hair surface that prevents ozone damage to the cuticle. Combing Force Combing studies were also used to demonstrate the ozone protection benefits of KERATEC IFP PE on the surface of the hair. The following 3g tresses of hair were used: Untreated baseline (control) hair fibers Hair fibers treated with conditioner base-rinse off Hair fibers treated with conditioner base + KERATEC IFP PE (1% as supplied)-rinse off

The tresses were exposed to ozone at a level of 20ppm for 2min. After pollutant exposure, tresses were washed, rinsed thoroughly and mounted in the apparatus (Instron 4204) wet. 10 combing strokes were performed and for each stroke a force/elongation graph was recorded. The force of each stroke was calculated and then the geometric mean result for all 10 combing strokes was obtained (Figure 3).

Combing Force of Ozone Exposed Hair Fibers

180 160 Average Force 140 120 100 80 60 40
Untreated Control Conditioner Base Conditioner Base + 1 1%Keratec IFP PE % Keratec IFP

p<0.05

Figure 3: Graph showing average combing force for ozone exposed hair fibers; 1% KERATEC IFP PE protected the hair and aided in conditioning

Increasing surface damage

KERATEC IFP PE

The graph shows that the hair was much harder to comb (169% of control) with the conditioner base suggesting there may be some interaction between the conditioner and the ozone. The tresses treated with conditioner base plus KERATEC IFP PE were significantly easier to comb (59% of the control) indicating that the KERATEC IFP PE protects the hair from ozone damage, making it easier and more manageable to comb. Sulphur Dioxide Tryptophan Fluorescence Measurement of tryptophan fluorescence was performed on hair following five exposures to SO2 gas. Each exposure consisted of placing the hair in a vessel containing sulphur dioxide at 40ppm and left overnight, by which time the level of sulphur dioxide had decayed to 0ppm. The same test protocol was used as in the ozone experiment. The following conditions were used: Untreated baseline (control) hair fibers Hair fibers treated with conditioner base - rinse off Hair fibers treated with conditioner base + KERATEC IFP PE (1% as supplied) - rinse off

Tryptophan Degradation of Hair Fibers Exposed to Sulphur Dioxide

Normalised Intensity of emitted light

100 95 90 85 80 75 Untreated control Conditioner base Conditioner base + 1% Keratec IFP 1 % Keratec IFP PE

Figure 4: Graph showing tryptophan degradation caused by exposure to SO2; 1% KERATEC IFP PE protected the hair fiber completely

Increasing surface damage

KERATEC IFP PE

The graph shows that exposure to sulphur dioxide reduces the emission intensity of the untreated control hair. The conditioner base again protected the hair to a certain extent, but KERATEC IFP PE provides 100% protection of tryptophan in the hair fiber. This again demonstrates that the KERATEC IFP PE film protects the cuticle layer from damage by acting as a protective layer and preferentially degrading itself, leaving the underlying cuticle unharmed. Combing Force Combing studies were also used to demonstrate the SO2 protection benefits of KERATEC IFP PE on the surface of the hair. The same test protocol was used as for the ozone experiment. The following 3g tresses of hair were used: Untreated baseline (control) hair fibers Hair fibers treated with conditioner base - rinse off Hair fibers treated with conditioner base + KERATEC IFP PE (1% as supplied) - rinse off

The tresses were placed in a vessel containing sulphur dioxide at 40ppm and left overnight, by which time the level of sulphur dioxide had decayed to 0ppm. After pollutant exposure, tresses were washed, rinsed thoroughly and the combing forces measured.

Combing Force of Sulphur Dioxide Exposed Hair Fibers

Increasing surface damage
450 425 Average Force 400 375 350 325 300 Untreated Control Conditioner Base Conditioner Base + 1%% Keratec IFP PE 1 Keratec IFP p<0.1

Figure 5: Graph showing average combing force for hair swatches exposed to SO2; 1% KERATEC IFP PE protected the hair and aided in conditioning

KERATEC IFP PE

It appears that the conditioner base plus KERATEC IFP PE decreases the force required to comb the hair (80% of control). The conditioner base is not significantly different from the control. These results show that the KERATEC IFP PE protects the hair from SO2 damage, providing a reactive coating to mop up the harmful pollutant and shield the underlying hair cuticle, making it easier to comb. Smoke Combing Force This experiment was carried out using the same test protocol as for the ozone experiment. The tresses were exposed to cigarette smoke in a 3 liter enclosed volume. A cigarette was smoked into this volume over 10 minutes and the tress was left to stand in the smoke for a further 20 minutes. This process was repeated a total of 8 times with treatment of the base conditioner and IFP/conditioner being repeated after 4 cycles. The combing forces were measured and are shown in the graph below (Figure 6).

Combing Force of Smoke Exposed Hair Fibers

Increasing surface damage
220 200
Average Force

180 160 140 120 100 80 60 Untreated Control Conditioner Base Conditioner base + 1% Keratec IFP 1 % Keratec IFP PE
p<0.06

Figure 6: Graph showing average combing force for hair fibers exposed to smoke; 1% KERATEC IFP PE protects and hair and aids in conditioning

KERATEC IFP PE

The data shows that with smoke exposed hair, the conditioner base treated hair required a lower combing force (68% of control) than the untreated hair indicating that it gave some protection against the smoke. The conditioner base containing KERATEC IFP PE gave significantly greater protection; the swatch required a much lower combing force (38% of control) compared to the untreated hair. From these results it appears that the KERATEC IFP PE protects the hair fibers from the many reactive species in smoke by forming a protective film which preferentially absorbs harmful species and then is washed away leaving the cuticle beneath in a more manageable and healthy condition.
1

Jackowicz and Pande, J. Soc. Cos. Chem., 44, pp 109-122 (1993).