Introduction. In vivo study Skin Adhesion of the Active. Dermoprotectyl cellular active. Dermoprotectyl cellular active

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1 Introduction Environmental and lifestyle factors can play a significant role in the aging of skin. The most common culprit is UV light, which causes free radical formation that may lead to major changes in the skin cells DNA, its collagen and elastin tissue, and its pigmentation. This photoaging process can cause wrinkles, hyperpigmentation and potentially serious skin conditions. UV protection is considered the first line of defense when preventing UVA and UVB rays from entering the skin. Research suggests that1: regular use of topical photoprotective agents significantly reduces lifetime UV exposure the most important factor is to begin regular daily use early in life, and there is good reason to expect that regular use of daily facial topical products containing UV filters, particularly if started early in adult life, will be translated into fewer visible signs of aging later in life In vivo study Skin Adhesion of the Active A Skin Adhesion Study was undertaken to quantify the adhesive properties of relative to equivalent amounts of unencapsulated UV filters. Solutions containing Dermoprotectyl cellular active or unencapsulated UV filters were applied to each forearm of 10 panelists using standard skin test protocols. After drying, one test area per arm remained untreated, a second test area was rinsed with 10 ml water, and a third was rinsed with 2 x 10 ml water. Once dry, the product residues were entirely removed from each test area with cotton buds drenched in 70 % ethanol and subsequently extracted for analysis. The amount of UV filter remaining was quantified by recording the UV absorbance at the absorption peak of each UV filter. UV protection alone isn t infallible however. The sun also emits visible light and infrared radiation, which have been shown to also be a source of skin damaging free radicals. Antioxidants can provide a second line of defense by helping to stop the harmful chain reaction that free radicals initiate. Providing overall sun protection therefore requires formulators to consider strategies that address both the first line and second line of defense. UV Absorption [%] (normalized to 0 washings) Skin Adhesion of Dermoprotectyl Cellular Active Dermoprotectyl cellular active is a sun protection ingredient which contains both the first line (UVA and UVB filter) and the second line of defense (antioxidants Vitamin C and Vitamin E). In vivo studies with show that: >> antioxidants are delivered into the epidermis to obtain high radical protection, and >> the UVA and UVB filter adhere firmly to the skin surface to achieve effective photo protection This dual action defense relies on two different delivery technologies and allows for reduced signs of aging that may result from sunlight exposure Before Washing 1 Washing 2 Washings Unencapsulated UV Filters 1. Diffey B.L., Institute of Cellular Medicine, University of Newcastle, UK is a dual action sun protection active for sophisticated formulations targeted to provide sunlight protection delivers antioxidants to the deeper skin layers to achieve high free radical protection affixes UV filters to the skin surface to provide high photo protection increases the skin s antioxidative protection demonstrates strong adhesive properties of the UV filter on the skin surface provides an SPF value dependent upon the use level of the product The information contained herein is offered without charge for use by technically qualified personnel at their discretion and risk. All statements, technical information and recommendations contained herein are based on tests and data that we believe to be reliable, but the accuracy or completeness thereof is not guaranteed and no warranty of any kind is made with respect thereto. Air Products and Chemicals,Inc., 2012 Pub. No GLB

2 Introduction Environmental and lifestyle factors can play a significant role in the aging of skin. The most common culprit is UV light, which causes free radical formation that may lead to major changes in the skin cells DNA, its collagen and elastin tissue, and its pigmentation. This photoaging process can cause wrinkles, hyperpigmentation and potentially serious skin conditions. UV protection is considered the first line of defense when preventing UVA and UVB rays from entering the skin. Research suggests that1: regular use of topical photoprotective agents significantly reduces lifetime UV exposure the most important factor is to begin regular daily use early in life, and there is good reason to expect that regular use of daily facial topical products containing UV filters, particularly if started early in adult life, will be translated into fewer visible signs of aging later in life In vivo study Skin Adhesion of the Active A Skin Adhesion Study was undertaken to quantify the adhesive properties of relative to equivalent amounts of unencapsulated UV filters. Solutions containing Dermoprotectyl cellular active or unencapsulated UV filters were applied to each forearm of 10 panelists using standard skin test protocols. After drying, one test area per arm remained untreated, a second test area was rinsed with 10 ml water, and a third was rinsed with 2 x 10 ml water. Once dry, the product residues were entirely removed from each test area with cotton buds drenched in 70 % ethanol and subsequently extracted for analysis. The amount of UV filter remaining was quantified by recording the UV absorbance at the absorption peak of each UV filter. UV protection alone isn t infallible however. The sun also emits visible light and infrared radiation, which have been shown to also be a source of skin damaging free radicals. Antioxidants can provide a second line of defense by helping to stop the harmful chain reaction that free radicals initiate. Providing overall sun protection therefore requires formulators to consider strategies that address both the first line and second line of defense. UV Absorption [%] (normalized to 0 washings) Skin Adhesion of Dermoprotectyl Cellular Active Dermoprotectyl cellular active is a sun protection ingredient which contains both the first line (UVA and UVB filter) and the second line of defense (antioxidants Vitamin C and Vitamin E). In vivo studies with show that: >> antioxidants are delivered into the epidermis to obtain high radical protection, and >> the UVA and UVB filter adhere firmly to the skin surface to achieve effective photo protection This dual action defense relies on two different delivery technologies and allows for reduced signs of aging that may result from sunlight exposure Before Washing 1 Washing 2 Washings Unencapsulated UV Filters 1. Diffey B.L., Institute of Cellular Medicine, University of Newcastle, UK is a dual action sun protection active for sophisticated formulations targeted to provide sunlight protection delivers antioxidants to the deeper skin layers to achieve high free radical protection affixes UV filters to the skin surface to provide high photo protection increases the skin s antioxidative protection demonstrates strong adhesive properties of the UV filter on the skin surface provides an SPF value dependent upon the use level of the product The information contained herein is offered without charge for use by technically qualified personnel at their discretion and risk. All statements, technical information and recommendations contained herein are based on tests and data that we believe to be reliable, but the accuracy or completeness thereof is not guaranteed and no warranty of any kind is made with respect thereto. Air Products and Chemicals,Inc., 2012 Pub. No GLB

3 In vivo study Dual Action Delivery 1. Dual Action Delivery of the Active To demonstrate the dual action delivery of sun protection by, studies were performed using 5D intravital tomography, a novel tool for noninvasive in vivo analysis of human skin. Using untreated skin as a baseline, a simple solution containing was evenly applied on the inner forearm of a panelist. The solution was allowed to dry. Then an emission at three different excitation wave lengths was measured after application. An overlay image (right) was taken at a skin depth of 30 m 30 minutes after application. As the image to the right suggests, Dermoprotectyl cellular active is able to deliver antioxidants (indicated in blue) deep into the skin while maintaining the UV filters (indicated in red) on the skin surface. 5D Intravital Tomography Image of Dermoprotectyl Cellular Active 30 μm 2. Dual Action Delivery of the Active in a Cosmetic Formulation To demonstrate the efficacy of the dual action delivery mechanism in a more complex cosmetic formulation, the experiment above was performed using in a sunscreen lotion. Again, 5D intravital tomography results suggest effective deposition of UV filters on the skin surface while achieving delivery of antioxidants deep into the skin. 5D Intravital Tomography Image of Dermoprotectyl Cellular Active used in a Sunlotion 30 μm

4 Ex vivo study Radical Skin Protection Factor (RSF) The RSF method can be used to quantify the UV-induced reactive oxygen species (ROS) damage in skin and to determine the protective effect of UV filters and antioxidants. High RSF values mean high protection against free radical damage. was applied on the epidermal side of a skin biopsy and allowed to penetrate for 15 minutes. For comparison, equivalent amounts of unencapsulated UV filters were applied in the same way. Prior to measurements, the biopsy was treated with a solution containing a radical indicator. After recording an initial baseline electron spin resonance (ESR) spectrum, the skin biopsy was irradiated with increasing doses of UV radiation, and the ESR spectrum was determined after each radiation step. Radical Skin Protection Factor Dermoprotectyl cellular active Unencapsulated UV filters RSF = Number of free radicals in untreated skin Number of free radicals in treated skin can provide a very high level of UV protection, with an RSF of >99 indicating nearly absolute radical protection. By comparison, equivalent amounts of unencapsulated UV filters show an RSF value of only 7.1.

5 SAP [%] (Normalized to untreated skin) Ex vivo study Skin Antioxidative Protection (SAP) The SAP method quantifies the efficacy of a cosmetic antioxidant formulation inside the skin. The skin is labeled with an ESR active test radical, the amount of which is reduced by the antioxidant system inside the epidermis and dermis. An effective substance, applied to the stratum corneum and able to penetrate, will increase the skin s antioxidative protection. A time dependent penetration profile of the tested substance can be obtained on skin biopsies labeled with a semi-stable free radical. The signal intensities are registered by ESR spectroscopy. (10% in water) was tested and compared to untreated skin. Skin Antioxidative Potential of Dermoprotectyl Cellular Active Time [min] can lead to a significant increase in the skin s antioxidative power after 10 minutes and 30 minutes penetration time in relation to the untreated skin.

6 Introduction Environmental and lifestyle factors can play a significant role in the aging of skin. The most common culprit is UV light, which causes free radical formation that may lead to major changes in the skin cells DNA, its collagen and elastin tissue, and its pigmentation. This photoaging process can cause wrinkles, hyperpigmentation and potentially serious skin conditions. UV protection is considered the first line of defense when preventing UVA and UVB rays from entering the skin. Research suggests that1: regular use of topical photoprotective agents significantly reduces lifetime UV exposure the most important factor is to begin regular daily use early in life, and there is good reason to expect that regular use of daily facial topical products containing UV filters, particularly if started early in adult life, will be translated into fewer visible signs of aging later in life In vivo study Skin Adhesion of the Active A Skin Adhesion Study was undertaken to quantify the adhesive properties of relative to equivalent amounts of unencapsulated UV filters. Solutions containing Dermoprotectyl cellular active or unencapsulated UV filters were applied to each forearm of 10 panelists using standard skin test protocols. After drying, one test area per arm remained untreated, a second test area was rinsed with 10 ml water, and a third was rinsed with 2 x 10 ml water. Once dry, the product residues were entirely removed from each test area with cotton buds drenched in 70 % ethanol and subsequently extracted for analysis. The amount of UV filter remaining was quantified by recording the UV absorbance at the absorption peak of each UV filter. UV protection alone isn t infallible however. The sun also emits visible light and infrared radiation, which have been shown to also be a source of skin damaging free radicals. Antioxidants can provide a second line of defense by helping to stop the harmful chain reaction that free radicals initiate. Providing overall sun protection therefore requires formulators to consider strategies that address both the first line and second line of defense. UV Absorption [%] (normalized to 0 washings) Skin Adhesion of Dermoprotectyl Cellular Active Dermoprotectyl cellular active is a sun protection ingredient which contains both the first line (UVA and UVB filter) and the second line of defense (antioxidants Vitamin C and Vitamin E). In vivo studies with show that: >> antioxidants are delivered into the epidermis to obtain high radical protection, and >> the UVA and UVB filter adhere firmly to the skin surface to achieve effective photo protection This dual action defense relies on two different delivery technologies and allows for reduced signs of aging that may result from sunlight exposure Before Washing 1 Washing 2 Washings Unencapsulated UV Filters 1. Diffey B.L., Institute of Cellular Medicine, University of Newcastle, UK is a dual action sun protection active for sophisticated formulations targeted to provide sunlight protection delivers antioxidants to the deeper skin layers to achieve high free radical protection affixes UV filters to the skin surface to provide high photo protection increases the skin s antioxidative protection demonstrates strong adhesive properties of the UV filter on the skin surface provides an SPF value dependent upon the use level of the product The information contained herein is offered without charge for use by technically qualified personnel at their discretion and risk. All statements, technical information and recommendations contained herein are based on tests and data that we believe to be reliable, but the accuracy or completeness thereof is not guaranteed and no warranty of any kind is made with respect thereto. Air Products and Chemicals,Inc., 2012 Pub. No GLB