GREENOXIA-3. Purpose for Dual-glow Balancing agent. 31, Geondudanji 1gil, Dongnae-myeon,

Transcription

1 GREENOXIA-3 Purpose for Dual-glow Balancing agent 31, Geondudanji 1gil, Dongnae-myeon, Chuncheon, Gangwon, Republic of Korea page1

2 GREENOXIA-3 is good for Dual-glow Balancing For beautiful skin, skin glow is very important because it presents skin's health and vitality. Skin glow is consisted of 2 glows; shiny glow and moisture glow. If they are not balanced, It causes aging and other skin troubles, like dryness, acne, atopic disease and even aging and wrinkle problems. Dry & Baby skin has not enough sebum in their skin and has weak skin barrier to protect skin effectively. Dry skin has lower level of sebum. So their skin easily dried and has low skin elasticity as well as low skin recovery level. Babies have think skin layer than adult around 30% less because their skin isn't fully developed so produces the small amount of sebum. And it's not enough to protect skin as a skin barrier. GREENOXIA-3, as a dual-glow balance agent, it balances skin glow by promoting sebum production and moisturizing effect. With GREENOXIA-3's valuable effect, keep highly glowed skin from dryness and other skin troubles. [Patent No. : ] Cosmetic composition for improving skin barrier with the extract of Chicorium intybus and Poncirus trifoliate rafin / RADIANT applied a patent for baby care. This patent showed that as effective component, it increases lipid biosynthesis function and moisturizing factor to protect baby s skin lacking the ability of strengthening skin barrier which causes skin trouble, dehydration or atopic dermatitis. page2

3 PART Ⅰ General Information 1. About GREENOXIA Why is dual (oil-moisture) balance important? Dry Skin & Baby Skin 4. Tannin PART Ⅱ Technical Data 1. Explain of GREENOXIA Stability study Safety study 16 PART Ⅲ Efficacy Data 1. Cholesterol Biogenesis effect Moisturizing Effect 3. Clinical Test of dual glow PART Ⅳ Conclusion 1. Summary 2. Application page3

4 1. About GREENOXIA-3 GREENOXIA-3 is composed of 3 green vegetables extract; Chicory extract, Salary extract and Broccoli extract. Chicory Leaf Extract Common name Latin name INCI name Main Ingredient Biological Effct Chicory Cichorium intybus Cichorium intybus (Chicory) leaf Extract Tannin, Dietary fiber Moisturizing, Anti-oxidant Chicory is one of the popular healthy vegetables because it has useful ingredients and abundant dietary fibers. So it is effective to constipation and good for diet with low calorie. And it stimulates the immune system and alleviates intestinal diseases and harmful bacteria. Chicory acts in similar ways to the beneficial bacteria commonly can be found in yoghurt. Chicory contains Carotene, Vitamin B2, Vitamin C, potassium, iron, etc which is good for skin beauty and also helps digestion and blood circulation. Healing power of Chicory These days, people enjoy chicory as a healthy juice not even for food such as salad, ice-cream, etc. It has detox effect good for diet or skin trouble problem. Chicory is one of the richest sources of vitamin A which is very useful for eyes. Chicory Tea is also popular as a substitute for coffee. It can be bought from many stores or can be made by yourself at home. It has abundant vitamin A, B, C, K and enhances absorption of calcium and strengthening the skin barrier. Also, stimulates the healing process of the skin and can be used to disinfect cuts. page4

5 Celery Extract Common name Celery Latin name Apium Graveolens L. INCI name Main Ingredient Biological Effect Apium Graveolens (Celery) Extract Tannin, Beta-carotene, Polyacetylene, Luteolin Anti-oxidant, Anti-irritation, Celery is an important source of conventional antioxidant nutrient, including vitamin C, beta-carotene. Celery has been considered to provide anti-inflammatory benefits because it contains polyacetylene which is considered one of the best defenders against inflammation. Other useful ingredient, luteolin, interferes with brain inflammation and inhibits the production Celery has detox effect of the body and helps refreshment and relax the body from stressful environment. Also, with vitamin C, it has effect on anti-aging and anti-tumor so that recommended to eat regularly for a long time. Woman uses Celery as a source of pack for their skin conditioning. Celery pack has lots of vitamin C that inhibits freckles, pigmentation and dark circles. It is good to pack 2times a week and give abundant moisture after clean the face. page5

6 Broccoli Extract Common name Latin name INCI name Main Ingredient Biological Effect Broccoli Brassica oleracea var. italica Brassica Oleraceaitalica (Broccoli) Extract Tannin, Flavonoids, Beta-Carotene Anti-oxidant, Anti-inflammation Broccoli, it s the most commonly eaten parts of broccoli are the flowering heads, which are shaped like a tree. Broccoli contains mineral, vitamin A, C, K and potassium, manganese. It recovers damaged skin and is well known for anti-oxidant effect that inhibits aging. As a one of super food, it inhibits disease related to cancer, blood, stomach and has lots of potassium that is essential for growth. In terms of unique organic compounds, broccoli is a rich source of flavonoids and various antioxidant compounds. Good for Baby Food Broccoli is a good source for baby food. Babies from 6 months to under 2 years eat baby food made by rice, vegetables, meats, etc. To provide abundant nutrients, moms make baby food with various ingredient depending on age and growth steps. Broccoli is popular as a green vegetable for baby food because it helps baby s digestion and prevents constipation. page6

7 2. Why is dual (oil-moisture) balance important? Necessary for Skin Protection When determine skin type or aging level, the important thing is how much moisture your skin has. Young or old is determined whether it has appropriate moisture depending on your ages not only ages. If the skin doesn t have appropriate moisture depending on age, it can be said to older than others. Compared normal skin and dried skin, dried skin has rough stratum of corneum and has less moisturizing protection than normal skin. Balance between oil and moisture There are 2 types of dried skin, Moisture-deficient skin and Lipid-deficient skin. When we have low level of moisture in skin, it s called moisture-deficient skin. Normally facial skin has low moisture because it ex exposed directly to outer factors. If the skin is deficient in NMF(Natural Moisturizing Factors), it loses moisture easily and feel dryness. Even it accelerates aging or wrinkle. Lipid-deficient skin has low level of sebum (sebaceous fats) in skin. During cleansing or other environmental factors, we loss valuable lipids and if we have not enough sebum, our skin can t inhibit water evaporation and form protect barrier. Skin conditioning is a first step for natural and glossy skin and for this, balance between oil and moisture very important. Who is keeping the skin from losing moisture? The answer is oil. Oil in skin acts as a protective layer and inhibits dry or wrinkle. This is why oil & moisture balance in skin care emphasized. page7

8 Key factor of Anti-Aging Lack of moisture from the skin cell causes skin aging. If you do not properly moisturize the skin, cell regeneration ability and skin elasticity is decreased and increase the wrinkle and aging effect. Not only moisture but also lack of oil causes dried skin and wrinkles. So the first condition for healthy and elastic skin is balance of oil and moisture. Everyone knows that easily happen to wrinkle in dried skin but most of the people usually have a lot of interest in moisture not oil. Oil inhibits evaporation of moisture and necessary for soft and silky skin. Skin itself has balancing power but as older its ability fall and accelerate aging with many external factors. GREENOXIA-3 has purpose of harmonious balance between oil and moisture and keep the skin from dry and moisture evaporation. GLOW TREND in Korea In recent years, bare face and natural makeup looks naked face is popular in Korea beauty market. Celebrities boast their beauty and be loved by many people with stylish passion, hair, make up, etc. Especially Women are envy their beautiful and glossy face and want to look like them. Their natural and glossy face make up became a trend rapidly and called glow skin. Glow skin means vital and smooth skin. Creamy smooth skin and shiny glow is a key point of glow skin. As a one of major beauty trend in Korea and this trend continually be loved because even general public can easily follow up their elastic and shiny glow skin care. With natural and glossy face make up, many cosmetics are launched in the market and many celebrities showed their cosmetics and make up for glossy face through beauty show program. Many famous beauty show deal with natural, shiny, glossy skin subjects and introduce related cosmetics and make up. For now, we easily hear shiny glow, glow skin, skin looks pottery, etc. Also, there are many cosmetics focused on glow skin and named glow essence, glow bb cream, glow mist, glow air-cushion as well as glow make up page8

9 3. Dry Skin & Baby Skin Dry skin and Baby skin has low level of sebum in their skin and weak skin barrier. Dry skin has not enough moisture and epidermal lipid (sebum) and Baby has weak and thin outer skin layer to protect their skin. <sebum> Sebum is an oily substance which sebaceous glands produces to protect skin from losing moisture Sebum forms protective barrier against water evaporation but if external factors overwhelms the ability of sebum, the skin will shrivel and crack. Sebum decreases with age and loses while washing; excessive bathing, scrubbing or harsh soap that dissolve the protective layer of sebum. When moisture content of the skin layer falls below 8 percent, it becomes dry, rough and cracked called moisture-deficient skin. Because facial skin is exposed directly it loses more moisture than other areas. The horny skin layer is hydrated by water from deeper skin layers and normal perspiration. In contrast to moisture-deficient skin, an increased moisture requirement can be a problem, lipid-deficient skin. During daily cleansing it is often overburdened by cleansing substances or soaps and loses valuable lipids. Babies have thin skin layer than adult around 30% less and fewer lipids (fats) in their skin. Due to thin epidermis of the skin, they have less sebum which protects the epidermis from dryness and dehydration and gets dryness easily. Even moms apply moisturizer to baby s skin, during the day it is easily removed so they need to treatment focused fundamentally. Baby s skin is very sensitive to external irritants as the barrier is not yet fully developed and easily hurts. They can t take action effectively to external irritants and dryness so that balance of moisture & oil also important for babies. page9

10 4. Tannins of GREENXOIA-3 Tannins are natural chemical called a polyphenol that are obtaining from Chicory, Celery and Broccoli. Tannins are well known for their anti-oxidant, anti-bacterial and antiinflammatory properties. It reduces free radicals in the body and protects skin tissue from harmful or inflammatory factors causing inflammation or cancer. As a strong anti-oxidant agent, it inhibits aging by reducing free radicals. Not only has strong anti-inflammatory effect but also strengthens skin tissue. According to advanced research, tannins enhance binding of collagen and glycosaminoglycan. As a result, tannin increases skin moisturizing activity. Phytochemical composition (g/100g dry weight) of GREENOXIA-3 Tannins of GREENOXIA-3 adjust balance between moisture and sebum. This environmentally responsible ingredient helps to reduce inflammation by limiting sebum oxidation and thus control excess sebum production. GREENOXIA-3 is an ideal ingredient to keep skin glow. Tannins of GREENOXIA-3 bring skin balance and skin elasticity for fresher, smoother and glossier skin. page10

11 1. Explain of GREENOXIA-3 Overall Procedure for the preparation of GREENOXIA-3 Material purchase and also certificate of origin & quality Grinding whole dried Chicory, Celery, Broccoli a) Extraction and then 1st Filtration Centrifugation (4500rpm, 30min) and then 2nd Filtration Measurement of dry content on 110 Produce of final product and sterilization Yes No Quality control Yes Product packaging ready for market a ; Extraction with 70% EtOH - Extraction time ; 12hr - Extraction temperature ; 20~25 - Filtration: paper filtration (pore size: 1 μm ) for remove precipitation and suspended particles page11

12 GREENOXIA-3 S p e c i f i c a t i o n INCI Name / Composition Appearance Odor Water Butylene Glycol Cichorium Intybus (Chicory) Leaf Extract Brassica Oleracea Italica (Broccoli) Extract Apium Graveolens (Celery) Extract Light yellow Liquid Typical ph Specific Gravity Identification test Heavy Metal Arsenic (As) Preservative Total Aerobic Count Dark green 20ppm 2ppm Non-added Less than 100 cfu/g E. coli Not detected Salmonella Not detected Storage condition Sealed containers should be Stored at a temperature of 10~30 (50~86 ), Quality might be affected after opening packing, please refer to MSDS for more information. After opening the drums, sterilization is no more guaranteed Expiration Date 2 years in sealed original packing, stored in due conditions page12

13 2. Stability study Stability to ph We have evaluated the stability of GREENOXIA-3 at different ph, temperature, ethanol, at 1% to 5% in final products. ph of GREENOXIA-3 solution was measured using the potentiometric method. Measurements were made at room temperature within a ph range of 2 to 10. ph V/V % ± 3% ± 5% ± + ; stable, ±; slightly unstable GREENOXIA-3 shows generally stability between 2 to 8 tested ph. Accordingly it allows the use of various formulations in final products. page13

14 Stability to temperature This study was made at the ph of the solution (ph close to 5.0) at temperatures ranging from -20 to 65, for 6 days. Test sample Temp 1day 2day 3day 4day 5day 6day GREENOXIA ; stable, ±; slightly unstable Temp V/V 30min 60min 120min 30min 60min 120min 30min 60min 120min 1% % % ; stable, ±; slightly unstable GREENOXIA-3 shows stability during 6 days under -20, 60 condition and within tested range of temperature 40 ~80 during 2 hours. Accordingly, it allows the use of any formulations in final products. page14

15 Stability in the presence of ethanol The study of solubility in various water/ethanol mixtures was made at room temperature at the ph of the solution (ph close to 5.0) (V/V) Ethanol / H2O (V/V) 10/90 30/70 50/50 70/30 90/10 1% % ± 5% ; stable, ±; slightly unstable GREENOXIA-3 shows good stability within the tested range of ethanol content. It means you don t care about limitation of ethanol. Therefore, it allows the use in any formulations in final products. page15

16 3. Safety study Repeated insult patch test (RIPT) OBJECTIVE To determine the irritation and/or sensitization potential of a test material after repeated application under occlusive, semi-occlusive or open patches to the skin of human subjects. TEST MATERIAL GREENOXIA-3 10% solution in distilled water STUDY DATES This study was initiated on Sep 8 th, 2013 and was completed on Oct 11 th, PANEL SELECTION Panels of human subjects, male and female, randomly selected. No individuals were empanelled if they exhibited or had a history of acute or chronic dermatologic, medical, or physical conditions that could interfere with dermal scoring. TEST METHOD Patches were applied to the same site on Monday, Wednesday, and Friday for a total of 9 applications during the Induction period. The subjects remove the patches 24hours after each application. 24hour rest periods follow each removal. Prior to each reapplication, site(s) were graded for dermal irritation and sensitization Dermal scores 0 No visible skin reaction ± Barely perceptible erythema (minimal) 1+ Mild erythema (diffuse) 2+ Well defined erythema 3+ Erythema and oedema 4+ Erythema and oedema with vesiculation page16

17 Ten to 21 days after application of the final induction patch, challenge patch(es) are applied to previously unpatched sites, adjacent to the original induction patch sites. The challenge sites 24~72hours after application. REFERENCE Standard Operating Procedures, Clinical Trials , Repeat Insult Patch Test (RIPT) RESULT Induction Scores Challenge Scores Subject Number Subject Initials hr 48hr 72hr 1 KJW KYI KMH LJH JJS LBR KHS JIJ KHM KNI CONCLUSION Based on the test population of 10 subjects and under the conditions of this study, the GREENOXIA-3 10% solution identified did not demonstrate a potential for eliciting dermal irritation on sensitization. page17

18 Ames test for mutagenicity OBJECTIVE To screen for mutagens through the simple and inexpensive procedure that uses a bacterial test organism. It is a biological assay used in genetics, generally genetic toxicology, to test for mutagenic properties of a chemical compound. STUDY DATES This study was initiated on Sep, 9 th, 2013 and was completed on Sep 17 th, TEST ORGANISMS The test organism is a histidine-negative (his - ) auxotrophic strain of salmonella typhimurium that will not grow on a medium deficient in histidine unless a back mutation to his + (histidine-positive) has occurred. PRINCIPLE OF TEST METHOD It is recognized that the mutagenic effect of a product is frequently influenced by the enzymatic pathway of an organism, whereby non-mutagens are transformed into mutagens and vice versa when introduced into human system. REFERENCE K. Mortelmans, E. Zeiger., Mutation Research 455 (2000) 29-60, The Ames Salmonella. Microsome mutagenicity assay. RESULT Test samples TA98 His+ revertants/plate TA100 Spontaneous test NQO % GREENOXIA CONCLUSION Based on the test procedure and under the conditions of this study, 10% GREENOXIA-3 solution identified did not demonstrate a potential for mutagenicity. page18

19 UV absorbance measurement OBJECTIVE UV absorbance measurement test is to determine of phototoxicity of ingredient in vitro condition. The test can result of safety about phototoxicity by evaluating UV absorbance. PRINCIPLE OF TEST METHOD UVA(320~400nm), UVB(290~320nm), UVC(200~290nm), visible rays (400~700nm). The shortest wavelengths of UVC (and those even shorter; nm) are blocked by ordinary air. MATERIAL Sample Concentration GREENOXIA-3 0.1% GREENOXIA-3 0.5% GREENOXIA-3 1% RESULT CONCLUSION GREENOXIA-3 has no absorbance of UV and has no risk of phototoxicity. page19

20 MATERIAL SAFETY DATA SHEET WRITTEN BY : SungYong, Lee DATE : February. 19, 2014 SIGNATURE : 1. PRODUCT AND COMPANY IDENTIFICATION Product name : GREENOXIA-3 Product code : PEC489 Use : Raw material for cosmetic Manufacturer : RADIANT. 31 Geodudanji 1-Gil Dongnae-myeon Chucheon Gangwondo Republic of Korea Tel : Fax : radiantcmo@chol.com E mail Emergency call G2. COMPOSITION AND IMFORMATION OF INGREDIENTS Chemical Name : None Molecular weight N.A CAS Number (Butylene Glycol) EINECS (Butylene Glycol) INCI Name : Water Butylene Glycol Cichorium Intybus (Chicory) Leaf Extract Brassica Oleracea Italica (Broccoli) Extract Apium Graveolens (Celery) Extract Hazardous ingredients: None 3. HAZARDS Information provided on the health effects of this product is based on individual components. All ingredients are bound and potential for hazardous exposure as shipped is minimal. However, some vapours may be released upon heating and the end-user (fabricator) must take the necessary precautions (mechanical ventilation, respiratory protection, etc) to protect employees from exposure. Main hazards No Known Health hazards Health risks Experience shows no acute irritancy or toxic effects. Environment risks Handle the product with good working practice avoiding dispersion into the environment. Routes of exposure Inhalation, Ingestion, Skin/Eye contact. 4. FIRST AID Skin : Wash off with soap and plenty of water. Eyes: Immediately irrigate with water. Ingestion : Do not induce vomiting without medical advice. Inhalation : Move to fresh air in case of accidental inhalation of fumes from overheating or combustion. When symptoms persist or in all cases of doubt seek medical advice. 5. FIRE FIGHTING page20

21 NFPA: Health: 1 Flammability: 1 Reactivity: Unsuitable extinguishing media Flammable properties Flash 109 (228 F) (Closed Cup) point (test method): Flammable limits in air, % by Upper: No Information Lower: No Information volume: 394 (741 F) Carbon monoxide and butadiene. Autoignition temperature: Use alcohol type aqueous film forming foam for large fires. Use CO2 or dry Products of combustion: chemical for small fires. Extinguishing Media: Thoroughly decontaminate bunker gear and other fire-fighting equipment before re-use. Fire Fighting Environmental Water spray should be used to cool fire-exposed structures and vessels. Water Concerns: or foam may cause frothing. Water spray can be used to reduce the intensity of flames and to dilute spills to a non-flammable mixture. Keep personnel removed Fire Fighting Instructions: from and upwind of fire. If potential for exposure to vapors or products of combustion exists, wear full fire fighting turnout gear and NIOSH approved selfcontained breathing apparatus. Oxidizing chemicals may accelerate the burning rate in a fire situation. 6. ACCIDENTAL RELEASE Remove heat and sources of ignition. Drums and packing in danger should be cooled by pulverized water, as heating could provoke a rise in pressure with explosion or deflagration risks. Prevent entry into watercourses and pipeworks. Immediately mop up with suitable absorbant equipment for subsequent correct disposal according to current legislation. `7. HANDLING AND STORAGE Store in a dry place and away from light to insure the quality. Keep the drums well closed in a well aired place. Keep away from heat and sources of ignition. Do not smoke. In case of important heating of the liquid, there is a risk of formation of explosive mixtures with air. Risk of fire in case of contact with hot area, sparks or flames. 8. EXPOSURE CONTROL AND PERSONAL PROTECTION Respiratory protection Facial mask Hand Protection : Protective gloves Eye Protection : Glasses with air-tight protection Skin and body protection : Long sleeved clothing and safety shoes Engineering measures Heat only in areas with appropriate exhaust ventilation. Provide appropriate exhaust ventilation at machinery. 9. PHYSICAL AND CHEMICAL PROPERTIES Appearance: Liquid Colours: Light Yellow Odour: Typical Specific Gravity at 20 : Solubility in water: Soluble ph(10%soln.): page21

22 Identification Test Dark green Vapour pressure N.A. Vapour density N.A Preservative Non-added 10. STABILITY AND REACTIVITY Thermal decomposition: Distillation without decomposition at normal pressure, No thermal decomposition in case of correct storage and handling. Dangerous decomposition products: No dangerous decomposition products if storage and handling conditions are respected. In case of fire or thermal decomposition, release of carbon monoxide and carbonic anhydride. Dangerous reactions : Reacts violently with powerful oxidising agents. Hazardous decomposition products :May emit irritant/toxic vapour/fumes under fire conditions. 11. TOXICOLOGY Acute toxicity : No toxical effect known Sensitisation : Not sensitizing Inhalation : Inhalation is possible only under aerosol conditions, and vapour/fumes might be irritant/toxic. 12. ECOLOGICAL INFORMATION No ecologic effect known 13. DISPOSAL Recommended method to dispose the product without danger: Dispose in accordance with the current legislation preferably using high temperature incineration or In a biological purification station in accordance with the current legislation. 14. TRANSPORT INFORMATION Not dangerous for transport. (ROAD-RAIL, SEA, AIR) 15. REGULATORY INFORMATION Labelling according E.E.C. directives : Not submitted to labelling 16. OTHER INFORMATION This information is furnished without warranty, except that it is accurate to the best knowledge of RADIANT INC. The data on this sheet relates only to the specific material designated herein. page22

23 1. Cholesterol Biogenesis Effect Linoleic acid as a positive control Linoleic acid is a component of the cell membrane in vivo and plays an important role. As an essential fatty acid, it produces neutral fat such as Triglyceride, a good fatty acid for acting epidermal lipid to protect and inhibits water evaporation. Because of this efficacy, it is popular in a beauty market. <Mechanism of Linoleic acid> Linoleic acid PPAR-γ expression Nucleus Tryglyceride Lipogenesis Cholesterol Biogenesis page23

24 Cholesterol Biogenesis Effect of GREENOXIA-3 OBJECTIVE To investigate the effect of GREENOXIA-3 on lipogenesis in the human sebocyte cell line SZ95 in vitro. We measured the lipid droplet by Oil Red O staining, total cholesterol assay. And we also measured the expression level of PPAR-γ by PCR analysis. STUDY DATES This study was initiated on September 9 th, 2013 and was completed on October 11 th, 2013 TESTING MATERIALS GREENOXIA-3 Test concentration 1%, 2% GREENOXIA-3 TESTING METHOD Oil Red O staining - The Oil Red O Method is a common staining technique used for demonstrating lipids in tissue. It is performed on fresh frozen sections, as alcohol and xylene dissolves the lipids that are in the tissue. Oil Red O has largely replaced many Sudan Dyes that were used for demonstrating lipids, such as Sudan III and Sudan IV, because Oil Red O provides a deeper red color that is easier to see. Total cholesterol assay Total cholesterol assay measure the total cholesterol within serum, plasma, cell lysate, or tissue samples. The assays will detect total cholesterol (cholesteryl esters plus free cholesterol) in the presence of cholesterol esterase or only free cholesterol in the absence of the esterase enzyme. Expression of Reverse transcription-polymerase chain reaction (PCR) - Quantification was achieved by measuring light absorbency at 260 nm. In cases in which either total RNA quality or quantity was not sufficient to allow further analysis, the extraction procedure was repeated, using frozen tissue samples. The expression of PPAR-γ1 and PPAR-γ2 was assessed by quantitative RT-PCR. page24

25 RESULT [Oil red O staining] Lipogenesis in the human sebocyte 2% GREENOXIA-3 1% GREENOXIA-3 Linoleic acid Control [Total cholesterol assay] page25

26 [Expression of PPAR-γ by RT-PCR] PPARγ/β-Actin mrna ratio (% of control) 2% GREENOXIA-3 1% GREENOXIA-3 Linoleic acid Control CONCLUSION [Oil red O staining] To protect skin from dryness and other external factor, it needs proper lipogenesis in human skin. Sebocyte, one of stimulating lipogenesis factor can be found with Oil Red O staining on skin cell. From the result, GREENOXIA-3 showed high absorbance of Oil Red O as dose dependent manner compared with positive control, Linoleic acid. We can estimate that GREENOXIA-3 increases lipogenesis activity. [Total cholesterol assay] C holesterol is one of composition of epidermal lipid. From the study, we investigated total cholesterol contents in GREENOXIA-3. GREENOXIA-3 increase cholesterol contents as dose dependent manner and improving epidermal lipid in skin layer. [Expression of PPAR-γ by RT-PCR] PPAR-γ regulates the triacylglycerol synthesis and secretion. From a result, we checked PPAR-γ expression level by RT-PCR and GREENOXIA-3 showed competitive effect compared to Linoleic acid, a positive control. page26

27 Hyaluronan Content (% of control) Moisturizing effect OBJECTIVE Hyaluronic acid(ha) is an anionic, non-sulfated glycosaminoglycan distributed widely throughout connective, epithelial, and neural tissues. HA is produced mainly by fibroblasts. And also it is powerful moisturizing agent. By checking hyaluronate level, we evaluate moisturizing effect of GREENOXIA-3 STUDY DATES This study was initiated on August 26 th, 2013 and was completed on September 6 th, 2013 TESTING MATERIALS GREENOXIA-3 Test concentration 1, 2% GREENOXIA-3 TESTING METHOD Hyaluronic acid assay HA(Hyaluronic acid) is produced mainly by fibroblasts and other specialized connective tissue cells. Properly diluted serum or plasma and HA reference solutions are incubated in HABP-coated microwells, allowing HA present in samples to react with the immobilized binding protein. After the removal of unbound serum molecules by washing, HABP conjugated with horseradish peroxidase solution is added to the micro wells to form complexes with bound HA. Following another washing step, a chromogenic substrate of tetramethylbenzidine and hydrogen peroxide is added to develop a colored reaction. The intensity of the color is measured in optical density (O.D.) units with a spectrophotometer at 450nm. RESULT Control Linoleic acid 1% 2% GREENOXIA-3 CONCLUSION This is a study to check Hyaluronic acid (HA) content in GRRENOXIA-3. From a result we checked that Hyaluronic acid is enrich in GREENOXIA-3 as dose dependent manner and high contents 2 times to 3 times more compared control. By checking hyaluronic acid, we evaluate moisturizing effect of GREENOXIA-3. page27

28 Clinical Test of Dual glow OBJECTIVE Balance between moisture and sebum is important to keep good skin conditioning and when there is balance with 2 factors, skin look bright and glossy. To estimate shiny glow effect and moisture flow effect, check moisture level and sebum level of people who have dry skin STUDY DATES This study was initiated on August 26 th, 2013 and was completed on September 26 th, 2013 TEST METHOD Applicant : 20 people who have dry skin Period : 4 weeks Test material : 3% GREENOXIA-3 Test : Moisture level, Sebum level RESULT CONCLUSION To investigate shiny & moisture glow effect of GREENOXIA-3, checks sebum index and moisture index of 20 people for 4weeks. As a result, GREENOXIA-3 increases the level of sebum & moisture. Almost applicants who have dry skin feel smooth and supple during the clinical test. page28

29 [Reference] Reported by COURAGE+KHAZAKA GmbH This result showed normal healthy skin condition at internal conditions (20 C, 40-60% air humidity). From this result, you can know which skin type you have. The Moisture Measurement by Corneometer, CM825 probe Skin Type Inner forearm Very dry < 30 Dry Sufficiently moisturized > 45 The Sebum Measurement by Sebumeter, SM815 probe Skin Type Forehead, t-zone, scalp Hair Cheek, Eyelid, temple Dry, less sebum < 100 < 40 < 70 Normal Oily > 220 > 100 > 180 page29

30 SUMMARY When compared with healthy skin, dry & baby skin has thin and weak skin glow. Low sebum level in skin layer can t act as a skin glow properly and loses moisture easily. It causes wrinkle and makes look older. This is why dry and baby skin needs dual-glow balance. <Mechanism of GREENOXIA-3> < GREENOXIA-3 improves outer skin glow by increasing synthesis of sebum and brings glossy & hydrated skin (HA: Hyaluronic acid, TG: Triglyceride) > page30

31 APPLICATION GREENOXIA-3 can be applied to various kinds of cosmetics like baby care, facial care, and body care including shower gel, body lotion, hand cream. And specially can be applied as a specific treatment for dryness. page31