Sunday, 29 June 2014

Dehydroascorbic acid (DHA)(*)(Vitamin C oxidized) concentration of 4%, a potential of next generation of home-made facial skin solution

By Kyle J. Norton
Health article writer and researcher; Over 10.000 articles and research papers have been written and published on line, including world wide health, ezine articles, article base, healthblogs, selfgrowth, best before it's news, the karate GB daily, The Alan Hopkinson Daily, etc,.
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                 Dehydroascorbic acid (DHA)(*)(Vitamin C oxidized)
                  

 (*) http://authors.library.caltech.edu/11677/1/BORjbc37a.pdf (The oxidation of ascorbic acid and its reduction in vitro and vivo)

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Abstract
Introduction
Skin aging is one of the most visible ageing processes that occur constantly in our skin organ. Many ingredients from synthetic and natural sources have been proven in studies to have certain effects on skin aging. In the comparison of  5 ingredients to test for their effectiveness and improvement of facial skin such as wrinkles, roughness, appearance of fine lines, lack of elasticity, black and white heads, acne scars, aging spots, pigmentation, and aging eye bags:
Synthetic vitamin C oxidized has been used in some cosmetic products but its effectiveness of being used alone with different concentrations has not been known.
Green tea and Vitamin E has shown to effectively improve aging skin and delayed aging progression by many studies.
Method of Experimentation
60 Patients recruited with skin disorders or aging such as wrinkles, roughness, appearance of fine lines, lack of elasticity, black and white heads, acne scars, aging spots, pigmentation, and aging eye bags were divided into 5 groups:
Group1 with 36 patients were tested for synthetic vitamin C oxidized with concentrations of 2%, 3% and 4% respectively.
Group 2 and 3 had 12 patients each and were for the effectiveness of Green tea and Vitamin E.
Patients must also agree during the study that they will not to use any other topical products (such as including moisturizers, sunscreens, and fragrances, make-up and any facial procedures such as peels, facials, microdermabrasion, and injection of botulinum toxin type A or dermal fillers) but the testing solution. The study also examines the patient with any cosmetic surgery and/or medication use of which may interfere with the study, as well as history of facial skin diseases. All patients are also required to sign an informed consent. Solutions of synthetic vitamin C oxidized and green tea were remixed in order to maintain their refreshness and concentration levels.
 Patients were instructed to apply the provided solution on their facial skin for a duration of 12 weeks. Facial skin examinations were conducted and recorded every 4 weeks.
Results
All patients completed the trial. Topical administration of synthetic vitamin C oxidized 4% showed a significant improvement in smoothing of the skin, skin elasticity, firming, disappearance of black and white heads, liver spots, pigmentation, and healing acne and acne scars, followed by 3% and 2% respectively, but an insignificant effect on aging eye bags and fine lines in week 12 compared with the baseline.
Green tea showed a minimum significance for skin smoothing, skin elasticity, firming and other skin parameters.
Vitamin E gave a mild result in the stimulation of collagen production in the areas surrounding the eye, skin firming, smoothing and elasticity as well as reduction of wrinkles and fine lines.
Conclusion
Treatment of different concentrations of synthetic vitamin C oxidized exhibited a significant efficacy in overall appearance of skin parameters such as improvement of skin smoothing, elasticity, firming, disappearance of black and white heads, liver spots, pigmentations, and healing acne and acne scars through its antioxidant effect, probably through anti-inflammatory and skin absorption pathways, in doses, skin aging progression and durative depending manner when compared to other groups.
Green tea may be a potential home-made cosmetic product used to improve skin parameters and maintain healthy skin, as well as delay the progression of aging.
Vitamin E offers solution for the aging population with the reduction of wrinkles, aging eye bags and fine lines.
Since the experimental group is small in nature, further studies with a larger sample size are necessary to improve the ingredient’s efficacy.
DR. Leung said “Anti-aging should start at an early age.” The use of a topical solution is necessary to prevent acne, black and white heads, and other skin disorder parameters that may induce permanent damage to the skin. Maintaining healthy levels of collagen is the right step to inhibit skin roughness, scaliness, wrinkles and fine lines, aging pigments, etc. I will not be surprised to see many people who look 10 - 20 years younger than their biological age, 30 years from now.
Introduction
Skin aging is one of the most visible aging processes that occur constantly in our skin. According to the Clinical Centre of Nis, certain plant extracts may have the ability to scavenge free radicals to protect the skin matrix through the inhibition of enzymatic degradation, or to promote collagen synthesis in the skin, and affect skin elasticity and tightness (a). Another study suggested that free radicals induced domino effects in production of reactive oxygen species and can react with DNA, proteins, and fatty acids causing oxidative damage and impairment of the antioxidant system, leading to injuries, damage of the regulation pathways of skin causing wrinkles, roughness, appearance of fine lines, lack of elasticity, and de-pigmentation or hyperpigmentation marks (b).
Walking through the drug and commercial cosmetic stores, one can see hundreds of anti-aging creams and products displayed. Many of them have also gone through certain studies for its efficacy with advertisement through media and celebrities. In fact, according to statistics, Global Anti-Aging Products Market will be worth $291.9 Billion by 2015.
Searching through the key word of Dehydroascorbic Acid (Oxidized Vitamin C, DHAA) and skin has yielded no related studies or clinical trials. It may be the result of the solution having no commercial value as it can be made by any cosmetic purchaser, or the studies indicate that L-ascorbic acid must be formulated at pH levels less than 3.5 to enter the skin with maximal concentration of 20% for optimal percutaneous absorption. But according to Dr. Douglas Q. Kitt, Dehydroascorbic Acid (DHAA) permeates stratum corneum at a rate up to 12 times faster than AA. This supports the concept that lower concentrations of DHAA in topical preparations can enhance skin vitamin C levels with less potential side effects (c).
Ingredient 1: Dehydroascorbic Acid (DHA) (vitamin C supplement oxidized form)
Vitamin C, also known as L-ascorbic acid, is a water-soluble vitamin found in fresh fruits, berries and green vegetables. It is best known for its free radical scavenging activity and regenerating oxidized vitamin E for immune system support. In skin aging, the vitamin may improve solar radiation protection and epidermal aging (1) through production of collagen due to its antioxidant activity (2)(3).
Epidemiological studies linking vitamin C in prevention of skin damage and aging have produced some certain results (3a)(3b)(3c), but the large sample size and multi-centred studies are necessary to validate its effectiveness. Vitamin C oxidized form or dehydroascorbic acid (DHA) processed antiviral and virucidal effects (23) showed to prevent H2O2-induced cell death by increasing the GSH levels mediated by the GPx and GR activities and PPP (17), and regulated neuronal energy metabolism, through facilitating the utilization of glucose via the PPP for antioxidant purposes (18) by increasing antioxidant potential in the central nervous system (19). Dehydroascorbic acid (DHA) may be a potential anti-cancer agent to treat aggressive cancers (20)(21). Dr. Toohey at the Cytoregulation Research said "rapidly-dividing tumour cells make unusually large amounts of homocysteine thiolactone and that administered dehydroascorbic acid enters the cells and converts the thiolactone to mercaptopropionaldehyde which kills the cancer cells" (22).
1. Plasma levels of vitamin C L-ascorbic acid is known for its effects on skin-whitening and against the anti-oxidation causes of skin aging. During the aging process, levels of vitamin C was found to slowly deplete according to the Tokyo Metropolitan Institute of Gerontology in the measurement of plasma and urine of C57BL/6 male mice during 3 to 30 months of aging(4). Therefore, restoration of physiological levels of vitamin C inside the cells might improve the lysosomal degradation (protection of cell from the degradative enzymes through protection of the cytosol) in the outflow pathway cells and prevent the pathogenesis of glaucomadegrade proteins(5).
Unfortunately, some researchers suggested that higher levels of circulating vitamin C may not provide protection against incident radiographic knee OA, and be associated with an increased risk of knee OA(6) and the 1Panorama Research Institute and Regenerative Sciences Institute, insisted "careful attention to individual and family medical history and personal genomic data may prove essential to make wise dietary and supplement choices to be combined with exercise”(7).
2. The effects
According to the Minghsin University of Science and Technology, in doses of a dependent manner, the concentration of L-ascorbic acid induced absorption of the collagen solution in exhibition of smoothing wrinkles and clear up spots(8). Ascorbic acid (AA) is essential in stimulating collagen gene expression. In type 1 and type 4 collagen and SVCT2, the vitamin was found to enhance the expression of type 1 and type 4 collagens and SVCT2 mRNA in cultured human skin fibroblasts at 100 μM AA placed every 24h for 5 days to prevent depletion(9). The Chiang Mai University study also supported the effects of ascorbic acid in the anti-aging process through exhibition pro and active MMP-2 inhibitory(10). Other studies suggested the combined vitamins, including vitamin C in a single formulation had a slightly lower degradation rate and more stable formulations as compared to different preparations containing only one of the vitamins(11)(12). Amazingly, in vivo, application of vitamin C showed a significant reduction of oxidative stress in the skin, an improvement of the epidermal-dermal microstructure and a reduction of fine lines and wrinkles in aged skin within a relatively short period of time of product application(13). The Bruce and Associates study also insisted the effectiveness of vitamin C application over a 12 week period as the vitamin enhanced the overall intensity of pigmentation, fine lines and wrinkles, tactile roughness, and laxity with a 100% satisfaction of overall appearance of the tested subjects’ skin(14).
1. Skin roughness and Scaliness
Skin roughness due to aging is a result of dead skin cells shedding more slowly, and building up in the upper layer of skin causing skin complexion to become rough and dull. According to the University of California at San Francisco, they are the result of aging and anatomic site of which demonstrated a significant influence on skin roughness and scaliness(25). Another study suggested that frictional properties of skin are dependent on more than water content or non-apparent sweating and the role of sebum secretion may be one possible factor(26), or it may be result of depletion of surface lipid content(27).
2. Wrinkles
Wrinkles are also associated with aging, hormonal status, smoking, and intercurrent disease according to the study by the University of California-San Francisco(28). Some researchers suggested that vitamin C may play an important role in collagen production due to its antioxidant properties(28). Another study indicated a positive anti-wrinkle effect through consumption of a mixture of soy isoflavones, lycopene, vitamin C, vitamin E, and fish oil(29).
3. Skin elasticity and firming
Skin sagging is a result of a loss of collagen and elastin of the skin due to aging in combination with the dreaded pull of gravity. The study of a novel dietary supplement (Imedeen Prime Renewal) including soy extract, fish protein polysaccharides, extracts from white tea, grape seed and tomato, vitamins C and E as well as zinc and chamomile extract, showed to inhibit forehead, periocular and perioral wrinkles, mottled pigmentation, laxity, sagging, under eye dark circles, and overall appearance. After 6 months treatment, it was suggested that the formula may provide improved condition, structure and firmness of the skin in post-menopausal women(30). Other studies insisted that since vitamin C (Vit. C) in the form of L-ascorbic acid (Asc) can accelerate wound healing(33)(32) and protect fatty tissues from oxidation damage(33), it may play an integral role in collagen synthesis of reducing wrinkles and skin sagging(34)(35)
5. Liver and aging spots
Liver spot is a condition of brown or black spots appearing on the surface of the skin due to aging and exposure to ultraviolet radiation from the sun. According to skin care guides, high concentration of Vitamin C may improve the skin and lighten the spots(36). Other articles insisted that liver spots may be the result of a deficiency of vitamin C(37). According to the Duke University Medical Center, the application of topical solutions containing vitamins C and E provided protection for human skin against damage caused by ultraviolet radiation(38)(39)(40)(41).
6. Pigmentation
Skin pigmentation disorder is a result of damage most likely caused by UV sunlight, or unhealthy cells due to aging affecting the production of melanin. Vitamin C topical and by iontophoresis are found effectively in aiding post-laser hyperpigmentation  or short and long term treatment (42) for melism (43). In an experiment where 29 females with melisma enrolled for iontophoresis, a vitamin C solution was applied to one side of the face, while distilled water was applied to the other side as a control, treatment with colorimeter site showed a significant decrease in the luminance value, (a photometric measure of the luminous intensity per unit area of light) compared to that of the control site (44).
8. Acne and Acne scars
According to Dr. GEORGE E. MORRIS, M.D., in a study of 60 patients with acne being given 8 oz of citrus juice twice daily and vitamin C in 3 gm a day, after 4 months 43 showed improvement, 10 failed to show improvement and 7 did not return for follow up(45). Some articles showed that vitamin C may be effective in the reduction of the formation of acne scars through it’s anti-inflammatory and free radical scavenger activities(46), but no study has been found through searching on PubMed.

9. Back and White heads
Black and white heads are the result of a skin pore becoming clogged with sebum from the body's natural oil and becoming black heads if oxidized. No study has been found specifically for the key word vitamin C and back and white heads on PubMed.

10. Fine Lines
No study and been found
Ingredient 2: Vitamin E
 Vitamin E is a fat soluble vitamin consisting of eight different variants (alpha-, beta-, gamma-, and delta-tocopherol and alpha-, beta-, gamma-, and delta-tocotrieno with varying levels of biological activity, found abundantly in corn oil, soybean oil, margarine, wheat germ oil, sunflower, safflower oils, etc.) which plays an important role in neurological functions and inhibition of platelet aggregation, regulation of enzymatic activity, free radical scavenger, etc.
Epidemiological studies linking vitamin E in reduced progression of aging have been inconclusive (47)(48)(49)(50).
The effects
According to the Universiti Kebangsaan Malaysia, in the stress-induced premature senescence (SIPS) model of human diploid fibroblasts (HDFs), γ-tocotrienol (GTT), a vitamin E variant showed to delay cellular senescence of human diploid fibroblasts through the inhibition of intrinsic mitochondria-mediated pathway which involved the regulation of pro- and anti-apoptotic genes and proteins (51) and prevented cellular aging in human diploid fibroblasts (52), probably through modulating gene expression(53) and protecting against oxidative stress-induced cellular ageing by modulating the telomere length possibly via telomerase(54). The molecular mechanism of tocotrienol-rich fraction (TRF) study insisted that the variants protect against H(2)O(2)-induced oxidative stress in human skin fibroblast culture by modulating the expression of skin aging in COL I and COL III genes(collagen gene) with a concomitant increase in the rate of total collagen synthesis(55). According to the National University of Malaysia, alpha-tocopherol (ATF), another variant of vitamin E protected against H(2)O(2)-induced DNA damage and telomere shortening of normal human skin fibroblast cells derived from young and old individual donors(56) and topical administration of alpha-tocopherol (ATF), in medium doses, may provide some degree of photoprotection against UVA(57). Some researchers suggested that the potent of vitamins in reduced aging progression is a result of the balance between various skin antioxidants of vitamin E and C, through new delivery systems, in delivering vitamins C and E into the skin simultaneously(58). Other antioxidants that improve skin health include vitamin E, certain flavonoids and the carotenoids, β-carotene, lycopene, and lutein. These may also be effective through protection against inflammation, oxidative stress, and breakdown of the extracellular matrix(59). In a study of Indian women with signs of facial aging, the composition of niacinamide, panthenol, and tocopheryl acetate significantly reduced the appearance of hyperpigmentation, improved skin tone evenness, and lightened the appearance of the skin in comparison to the control through 6 weeks(60).



Ingredient 3: Green tea 
Green tea contains more amounts of antioxidants than any drinks or food with the same volume, and is made from the leaves of Camellia sinensis. Green tea undergoes minimal oxidation during processing and is originated from China. Green tea has been a precious drink in traditional Chinese culture and is used exceptionally in socialization for more than 4000 thousand years. Because of their health benefits, they have been cultivated for commercial purposes all over the world.

According to Popular Herbs - Green Tea(64), the leaves have been found effectively for treatment of :
1. Depression(61)(62)(63)
(http://tavatearesearches.blogspot.com/2011/11/organic-tava-tea-researches-ingredients_6101.html)
2. External Anogenital Warts
http://tavatearesearches.blogspot.com/2011/11/organic-tava-tea-researches-ingredients_5385.html
3. Obstructive Sleep Apnea-Related Brain Deficits
http://tavatearesearches.blogspot.com/2011/11/organic-tava-tea-researches-ingredients_4023.html
4. Bad Breath
http://tavatearesearches.blogspot.com/2011/11/organic-tava-tea-researches-ingredient_8701.html
5. Nervous Tension
http://tavatearesearches.blogspot.com/2011/11/organic-tava-tea-researches-ingredient_8983.html
6. Gastrointestinal diseases
http://tavatearesearches.blogspot.com/2011/11/organic-tava-tea-researches-ingredient_4605.html
7. Human immunodeficiency virus (HIV)
http://tavatearesearches.blogspot.com/2011/11/organic-tava-tea-researches-ingredient_7225.html
8. Immune system
http://tavatearesearches.blogspot.com/2011/11/organic-tava-tea-researches-ingredients_8786.html
9. Antimicrobial activities
http://tavatearesearches.blogspot.com/2011/11/organic-tava-tea-researches-ingredients_14.html
10. Periodontal Health
http://tavatearesearches.blogspot.com/2011/11/organic-tava-tea-researches-ingredient_14.html
11. Cognitive Activities
http://tavatearesearches.blogspot.com/2011/11/organic-tava-tea-researches-ingredient.html
12. Neurodegenerative diseases
http://tavatearesearches.blogspot.com/2011/11/organic-tava-tea-researches-ingredients_159.html
13. Cholesterol
http://tavatearesearches.blogspot.com/2011/11/organic-tava-tea-researches-ingredients_8751.html
14. Osteoporosis
http://tavatearesearches.blogspot.com/2011/11/organic-tava-tea-researches-ingredients_7779.html
15. Arthritis
http://tavatearesearches.blogspot.com/2011/11/organic-tava-tea-researches-ingredients_6638.html
16. Stroke
http://tavatearesearches.blogspot.com/2011/11/organic-tava-tea-researches-ingredients_7408.html
17. Antioxidant Activity
http://tavatearesearches.blogspot.com/2011/11/organic-tava-tea-researches-ingredients_3373.html
18. Diabetes
http://tavatearesearches.blogspot.com/2011/11/organic-tava-tea-researches-ingredients_5800.html
19. Weight Loss
http://tavatearesearches.blogspot.com/2011/11/organic-tava-tea-researches-ingredients_13.html

20. Skin surface parameters
The study of green tea and lotus extract, the two cosmetic ingredients used in active formulations for the treatment of facial wrinkles, showed a potential influence on skin surface parameters through their diverse anti-oxidant constituents(65). In support of the above, the Korea University indicated that green tea extract after tannase treatment showed to enhance anti-wrinkle activities through its antioxidant activity(66). Its polyphenol chemical constituent also found effective in reduction of skin roughness(67), and positively affected skin elasticity, roughness, scaling, density, and water homeostasis(68) and  its (-)-epigallocatechin-3-gallate protected against UVA- and UVB-induced skin damage(69)(70). In treatment of acne and acne scars, according to the Islamia University of Bahawalpur, green tea and lotus treatment produced statistically more sound results than green tea alone in a 60-day treatment course in significantly reducing sebum secretions(71). The stable formulation (water in oil emulsion) containing 3% green tea (Camellia sinensis L) extract showed a statistically significant result for skin sebum production after long-term application(72).
Method
A total of 60 patients were recruited:
1. Group A contained 36 patients with ages ranging from 20 - 60 (20 Caucasians, 8 Africans, and 8 Asians; 24 females and 12 males) to test for the effectiveness in the difference of concentrations of oxidized vitamin C supplements (2000mg, 3000mg, 4000mg diluted into  100ml water) to yield 2%, 3% and 4% respectively).
2. Group B contained 12 patients with ages ranging from 20 to 60 (8 Caucasians, and 4 Asians; 8 females and 4 males) to test for the effectiveness of green tea ( 2g/100ml hot water solution) for skin facial parameters.
3. Group C contained 12 patients with ages ranging from 20 to 60 (7 Caucasians, 1 African, and 4 Asians; 10 females and 2 males) to test for the effectiveness of skin facial parameters of vitamin A.
                                   
                                   
Total Patients Study 

Ethnicity Gender Ages 20-30 Ages 31-40 Ages 41-50 Ages 50-60
35 Caucasians 45 females
15
15
15
15
15 Asians 15 males



10 Africans





Patients are eligible to enter this study only with an oily type of skin
1. Group with ages ranging from 20 - 30
Selection was based on those who had moderate to severe white and black heads on the nose, acne scars, and photo-pigmentation.                
2. Group with ages ranging from 30 - 40
Selection was based on those who had moderate to severe white and black heads with acne scars, stretch marks, skin roughness, black and white head on the nose, acne scars, and pigmentation and photo-pigmentation.
3. Group with ages ranging from 40 - 50
Selection was based on those who had moderate to severe white and black heads with acne scars, stretch marks, skin roughness, eye bags, wrinkles, black and white heads on the nose, acne scars, pigmentation and photo-pigmentation.
4. Group with ages ranging from 50 -60
Selection was based on those who had moderate to severe white and black heads with acne scars, stretch marks, fine lines, skin roughness, eye bags, wrinkles, black and white heads on the nose, acne scars, and pigmentation and photo-pigmentation.
Patients must also agree during the study that they will not to use any other topical products but the testing solution, such as moisturizers, sunscreens, fragrances, make-up, and any facial procedures such as peels, facials, microdermabrasion, and injection of botulinum toxin type A or dermal fillers. The study also examines the patient with any cosmetic surgery and medication use which may interfere with the study, as well as history of facial skin diseases. All patients are also required to sign an informed consent.

Initial examination
Based on a scale from 0-4 (0 - Not applicable, 1 - Minimum, 2 - Mild. 3 - Moderate, 4 - Severe)                           

Groups Ages of 20 -30 Ages of 31 -40 Ages of 41 -50 Ages 51 - 60
Acne
4
4
4
4
Acne scars
2
3
4
4
Skin roughness
0
2
3
4
Wrinkles
0
1
3
4
Elasticity
0
1
3
4
Scaling
0
1
3
4
Aging spots
0
1
3
4
Photo-pigmentation
0
2
4
4
Eye bags
0
1
2
4
Black and white heads
4
4
4
4
Fine lines
0
0
3
4
The study also examined the patient with any cosmetic surgery and medication use which may interfere with the study, as well as history of facial skin diseases. All patients are also required to sign a informed consent.
Patients are divided into 5 Groups to test for the effectiveness of:
1. Vitamin C supplements oxidized in concentrations of 2%, 3% and 4%.
2. Green tea 2g in water solution of 100mg.
3. Liquid vitamin E.
 

Vitamin C supplement oxidized

Concentration Ages 20 -30 Ages 31 -40 Ages 41 -50 Ages 51 -60 Total
2%
3
3
3
3
12
3%
3
3
3
3
12
4%
3
3
3
3
12
Total
9
9
9
9
36

             Vitamin C supplement oxidized with concentration 2%


Ages 20 - 30 Ages 31 - 40 Ages 41 - 50 Ages 51 - 60
Ethnicity
2C, 1AF
2C, 1A
2C, 1A
1C, 1A, 1AF
Gender
2F, 1M
2F, 1M
3F
2F, 1M


Vitamin C supplement oxidized with concentration 3%


Ages 21 - 30 Ages 31 - 40 Ages 41 - 50 Ages 51 - 60
Ethnicity
2C, 1A
3C
2A, 1AF
2C, 1AF
Gender
2F, 1M
2F, 1M
3F
2F, 1M


Vitamin C supplement oxidized with concentration 4%


Ages 20 - 30 Ages 31 - 40 Ages 41-50 Ages 51 - 60
Ethnicity
2C, 1A
2C, 1A
1C, 1A, 1AF
2C, 1AF
Gender
2F, 1M
2F, 1M
3F
2F, 1M

Green Tea Solution


Ages 20 - 30 Ages 31 - 40 Ages 41 - 50 Ages 51 - 60
# of Patients
3
3
3
3
Ethnicity
2C, 1AF
2C, 1A
2C, 1A
1C, 1A, 1AF
Gender
3F
2F, 1M
2F, 1M
2F, 1M


Vitamin E Liquid


Ages 20 - 30 Ages 31 - 40 Ages 41 - 50                   Ages 51 - 60
# of Patients
3
3
3
3
Ethnicity
2C, 1AF
2C, 1A
2C, 1A
1C, 1A, 1AF
Gender
3F
1F, 2M
2F, 1M
3F


Legend
M - Male
F - Female
C - Caucasians
A - Asian(s)
AF - African(s)

Treatment solution
1. Equipments contained a solution provided or mixed by patients as instructed. All solutions were refrigerated to protect its effectiveness.
2. Solution was applied twice per day, once in the morning and once in the evening.
Patients applied the solution first to the affected areas using the sponge path containing vitamin C and Green tea, and cotton stick containing vitamin A with little pressure to enhance the absorption for about 1 minute, then the whole face. Then patients would repeat this procedure once more.
The sponge path would then be cleared and the solution of vitamin C oxidized would be returned to the refrigerator.
The cotton would then be thrown away and the vitamin E liquid would be returned back to storage in room temperature.
Mixed green tea solution in every use.
The time of application takes less than 3 minutes depending on the severity of the oily skin.
Patients were also advised not to let the solution get into their eyes. If this did happen, patients were advised to use cold water to clean them out.

Outcome measure
The result would be measured 3 times:
1. The end of week 4
2. The end of week 8
3. The end of week 12
Patients were measured for the intensity of the applicable area in each age group and to check for efficacy of the solution by comparing to the base line.

Results:
Observation report
1. At the end of week 4
Measurement of improvement of the group with vitamin C oxidized supplements with concentration of 2% using a scale rating from 0 - 100% improvement


Groups Ages 20 - 30 Ages 31 - 40 Ages 41 - 50 Ages 51 - 60
Acnes
30
30
20
15
Acnes scars
20
20
15
10
Skin roughness
N/A
50
30
20
Wrinkles
N/A
30
15
5
Elasticity
N/A
30
30
20
Scaling
N/A
30
30
20
Aging spots
N/A
50
30
20
Photo-pigmentation
N/A
30
30
20
Eye bags
N/A
10
0
0
Black and white heads
40
40
15
10
Fine lines
N/A
0
15
5
At the end of weeks 4, there was no report of any adverse effects.

2. At the end of week 4
Measurement of improvement of the group with vitamin C oxidized supplements with concentration of 3% using a scale rating from 0 - 100% improvement

Groups Ages 20 - 30 Ages 31 - 40 Ages 41 - 50 Ages 51 - 60
Acnes
35
35
25
15
Acnes scars
35
25
15
10
Skin roughness
N/A
60
40
25
Wrinkles
N/A
40
15
5
Elasticity
N/A
45
45
30
Scaling
N/A
50
45
30
Aging spots
N/A
55
35
25
Photo-pigmentation
N/A
40
35
25
Eye bags
N/A
10
0
0
Black and white heads
50
50
30

15
Fine lines
N/A
0
15
5
At the end of week 4, there was no report of any adverse effects.

3. At the end of week 4
Measurement of improvement of the group with vitamin C oxidized supplements with concentration of 4% using a scale rating from 0 - 100% improvement

Groups Ages 20 - 30 Ages 31 - 40 Ages 41 - 50 Ages 51 - 60
Acnes
50
50
45
35
Acnes scars
50
45
30
20
Skin roughness
N/A
70
50
40
Wrinkles
N/A
50
55
15
Elasticity
N/A
70
55
40
Scaling
N/A
70
45
30
Aging spots
N/A
60
45
30
Photo-pigmentation
N/A
60
45
30
Eye bags
N/A
15
0
0
Black and white heads
50
60
40

25
Fine lines
N/A
0
15
5
At the end of week 4, there was no report of any adverse effects.

4. At the end of week 4
Measurement of improvement of the group with green tea solution of 2g diluted into 100ml of water using a scale rating from 0 - 100% improvement

Groups Ages 20 - 30 Ages 31 - 40 Ages 41 - 50 Ages 51 - 60
Acnes
10
5
0
0
Acnes scars
10
5
0
0
Skin roughness
N/A
50
30
20
Wrinkles
N/A
15
10
5
Elasticity
N/A
40
40
30
Scaling
N/A
40
30
20
Aging spots
N/A
50
20
15
Photo-pigmentation
N/A
30
25
20
Eye bags
N/A
0
0
0
Black and white heads
40
10
5

5
Fine lines
N/A
0
5
0
At the end of week 4, there was no report of any adverse effects.

5. At the end of week 4
Measurement of improvement of the group with Vitamin E liquids using a scale rating from 0 - 100% improvement

Groups Ages 20 - 30 Ages 31 - 40 Ages 41 - 50 Ages 51 - 60
Acnes
10
5
0
0
Acnes scars
10
5
0
0
Skin roughness
N/A
40
35
15
Wrinkles
N/A
40
35
15
Elasticity
N/A
40
30
20
Scaling
N/A
40
30
20
Aging spots
N/A
30
20
15
Photo-pigmentation
N/A
30
25
20
Eye bags
N/A
30
20
10
Black and white heads
40
0
0

0
Fine lines
N/A
35
25
15
At the end of weeks 4, there was a report of 4 patients with an adverse effect of lip sensation.

2. At the end of week 8:
Observation report
1. At the end of week 8
Measurement of improvement of the group with vitamin C oxidized supplements with concentration of 2% using a scale rating from 0 - 100% improvement


Groups Ages 20 - 30 Ages 31 - 40 Ages 41 - 50 Ages 51 - 60
Acnes
45
40
20
15
Acnes scars
35
25
20
15
Skin roughness
N/A
60
45
30
Wrinkles
N/A
40
20
10
Elasticity
N/A
45
35
25
Scaling
N/A
45
30
25
Aging spots
N/A
60
40
30
Photo-pigmentation
N/A
40
30
20
Eye bags
N/A
10
0
0
Black and white heads
40
45
20

10
Fine lines
N/A
0
15
5
At the end of week 8 there was no report of any adverse effects.

2. At the end of week 8
Measurement of improvement of the group with vitamin C oxidized supplements with concentration of 3% using a scale rating from 0 - 100% improvement

Groups Ages 20 - 30 Ages 31 - 40 Ages 41 - 50 Ages 51 - 60
Acnes
50
45
30
20
Acnes scars
45
40
20
15
Skin roughness
N/A
60
50
30
Wrinkles
N/A
40
20
10
Elasticity
N/A
60
50
40
Scaling
N/A
60
50
35
Aging spots
N/A
60
45
35
Photo-pigmentation
N/A
60
45
35
Eye bags
N/A
15
0
0
Black and white heads
50
60
35

25
Fine lines
N/A
0
20
15
At the end of week 8 there was no report of any adverse effect.

3. At the end of week 8
Measurement of improvement of the group with vitamin C oxidized supplements with concentration of 4% using a scale rating from 0 - 100% improvement

Groups Ages 20 - 30 Ages 31 - 40 Ages 41 - 50 Ages 51 - 60
Acnes
65
60
50
45
Acnes scars
60
55
35
25
Skin roughness
N/A
80
60
50
Wrinkles
N/A
65
55
25
Elasticity
N/A
85
60
35
Scaling
N/A
85
55
40
Aging spots
N/A
70
55
40
Photo-pigmentation
N/A
75
50
35
Eye bags
N/A
20
0
0
Black and white heads
50
75
50

40
Fine lines
N/A
0
25
15
At the end of week 8 there was no report of any adverse effects.

4. At the end of week 8
Measurement of improvement of the group with green tea solution of 2g diluted into 100ml of water using a scale rating from 0 - 100% improvement

Groups Ages 20 - 30 Ages 31 - 40 Ages 41 - 50 Ages 51 - 60
Acnes
10
5
0
0
Acnes scars
15
5
0
0
Skin roughness
N/A
60
40
30
Wrinkles
N/A
15
10
5
Elasticity
N/A
40
30
20
Scaling
N/A
50
40
30
Aging spots
N/A
55
25
15
Photo-pigmentation
N/A
40
35
25
Eye bags
N/A
0
0
0
Black and white heads
40
10
5

5
Fine lines
N/A
0
5
0
At the end of weeks 8 there was no report of any adverse effects.


5 . At the end of week 8
Measurement of improvement of the group with Vitamin E liquid using a scale rating from 0 - 100% improvement

Groups Ages 20 - 30 Ages 31 - 40 Ages 41 - 50 Ages 51 - 60
Acnes
10
5
0
0
Acnes scars
10
5
0
0
Skin roughness
N/A
50
40
20
Wrinkles
N/A
45
35
20
Elasticity
N/A
50
40
25
Scaling
N/A
50
35
25
Aging spots
N/A
40
30
25
Photo-pigmentation
N/A
35
25
20
Eye bags
N/A
40
25
15
Black and white heads
40
0
0

0
Fine lines
N/A
40
35
25
At the end of week 8 there was a report of 4 patients with an adverse effect of lip sensation.

Observation report:
1. At the end of week 12                               
Measurement of improvement of the group with vitamin C oxidized supplements with concentration of 2% using a scale rating from 0 - 100% improvement

Groups Ages 20 - 30 Ages 31 - 40 Ages 41 - 50 Ages 51 - 60
Acnes
55
45
25
15
Acnes scars
45
30
20
15
Skin roughness
N/A
65
50
35
Wrinkles
N/A
50
25
15
Elasticity
N/A
55
40
35
Scaling
N/A
55
40
35
Aging spots
N/A
65
50
35
Photo-pigmentation
N/A
45
35
25
Eye bags
N/A
15
0
0
Black and white heads
40
55
20

10
Fine lines
N/A
0
15
5
At the end of week 12 there was no report of any adverse effects.

2. At the end of week 12
Measurement of improvement of the group with vitamin C oxidized supplements with concentration of 3% using a scale rating from 0 - 100% improvement

Groups Ages 20 - 30 Ages 31 - 40 Ages 41 - 50 Ages 51 - 60
Acnes
60
45
30
20
Acnes scars
45
40
20
15
Skin roughness
N/A
60
50
30
Wrinkles
N/A
40
20
10
Elasticity
N/A
60
50
40
Scaling
N/A
60
50
35
Aging spots
N/A
60
45
35
Photo-pigmentation
N/A
60
45
35
Eye bags
N/A
15
0
0
Black and white heads
50
60
35

25
Fine lines
N/A
0
20
15
At the end of week 12 there was no report of any adverse effects.

3. At the end of week 12
Measurement of improvement of the group with vitamin C oxidized supplements with concentration of 4% using a scale rating from 0 - 100% improvement

Groups Ages 20 - 30 Ages 31 - 40 Ages 41 - 50 Ages 51 - 60
Acnes
70
60
50
45
Acnes scars
60
55
35
25
Skin roughness
N/A
80
60
50
Wrinkles
N/A
65
55
25
Elasticity
N/A
85
60
35
Scaling
N/A
85
55
40
Aging spots
N/A
70
55
40
Photo-pigmentation
N/A
75
50
35
Eye bags
N/A
20
0
0
Black and white heads
50
75
50

40
Fine lines
N/A
0
25
15
At the end of week 12 there was no report of any adverse effects.

4. At the end of week 12
Measurement of improvement of the group with green tea solution of 2g diluted into 100ml of water using a scale rating from 0 - 100% improvement

Groups Ages 20 - 30 Ages 31 - 40 Ages 41 - 50 Ages 51 - 60
Acnes
10
5
0
0
Acnes scars
15
5
0
0
Skin roughness
N/A
60
40
30
Wrinkles
N/A
15
10
5
Elasticity
N/A
40
30
20
Scaling
N/A
50
40
30
Aging spots
N/A
55
25
15
Photo-pigmentation
N/A
40
35
25
Eye bags
N/A
0
0
0
Black and white heads
40
10
5

5
Fine lines
N/A
0
5
0
At the end of week 12 there was no report of any adverse effect.

5 . At the end of week 12
Measurement of improvement of the group with Vitamin E liquid using a scale rating from 0 - 100% improvement

Groups Ages 20 - 30 Ages 31 - 40 Ages 41 - 50 Ages 51 - 60
Acnes
10
5
0
0
Acnes scars
10
5
0
0
Skin roughness
N/A
50
40
20
Wrinkles
N/A
45
35
20
Elasticity
N/A
50
40
25
Scaling
N/A
50
35
25
Aging spots
N/A
40
30
25
Photo-pigmentation
N/A
35
25
20
Eye bags
N/A
40
25
15
Black and white heads
40
0
0

0
Fine lines
N/A
40
35
25
At the end of week 12, there was a report of 4 patients with an adverse effect of lip sensation.
Efficacy and Patient satisfaction:
With a scale ranging from 0 - 4 (0: no effect, 1: minimum, 2: moderate, 3: good, 4 excellent effect)

Groups Vitamin C supplement oxidized Vitamin C supplement oxidized Vitamin C supplement oxidized Green tea Vitamin E liquids
Concentrations
2%
3%
4%


Ages 20 - 30
3.00
3.3
3.33
2.33
3.00
Ages 31 - 40
2.67
3.00
3.33
2.33
2.67
Ages 41 - 50
2.33
3.00
3.33
2.00
2.33
Ages 51 - 60
2.00
2.67
3.00
1.67
2.33
As the data indicates, patient satisfaction depended onto the concentration of the solution and the progression of aging. Most patients satisfied with the performance of vitamin C oxidized with a concentration of 4%. Green tea ranged last in the scores.

Tolerability
Through over 12 weeks of application with the designed solution, no adverse effect has been reported except for 4 patients complaining of lip sensation in the vitamin E group.

Discussion
1. Vitamin C oxidized showed the most efficacy in enhanced smoothness, firmness, and elasticity of the skin through stimulation the production of collagen. It also whitened skin and reduced scaliness, photo-pigmentation and aging pigmentation, aging spots, acne, acne scars. It was especially effective in the treatment of black and white heads when compared with other solutions probably through its antioxidant activities via inflammatory pathways, skin absorption. 4% concentration is the best but it had little effect in reduced wrinkles and fine line, regardless of concentrations.
a. For the age group 20 - 40, Dehydroascorbic acid (DHA)(vitamin C oxidized) with 2% concentration may be a better choice to maintain the collagen which will be lost gradually in aging.
b. For the age group 41 - 50, vitamin A is the best choice to reduce wrinkles and fine lines caused by aging, as well as enhancing the production of collagen for reduction of the scaliness of skin, photo-pigmentation and aging pigment, and aging spots. Apply the vitamin C concentration at 2% or 4% may be effective in reduced acne, acne scars, and black and white heads.
c. For the age group 51 - 60, proceed the same way as the age group of 41 -50. 

Would the higher concentration than 4% of Dehydroascorbic acid (DHA)(vitamin C oxidized) be more effective? The question is remained unknown as of today.

2. Vitamin E liquids showed a mild effect in enhanced smoothness, firming, and elasticity of the skin through stimulation the production of collagen, reduced wrinkles and fine lines especially in the forehead, nose-cheek areas, and at the end of the eyes, probably through its antioxidant effects through cell cycle division.

3. Green tea showed a minimum effect  in enhanced whitened skin and reduced scaliness, photo-pigmentation and aging pigmentation, aging spots, including smoothness, firming, and elasticity of the skin through stimulation of the production collagen.

The efficacy of the solutions indicated a logical thinking through  maintaining healthy skin by preventing the loss of collagen due to aging is the most effective way to slow down the aging progression. Vitamin C oxidized at 4% concentration offers the best treatment for this purpose. Logically, higher concentrations, such as 8% -12%  may provide us with more insight of this astonished result, but further study is necessary to rule out any adverse effect.
Since the sample is small, further studies with larger sample size and muti-centered studies are necessary to validate the claims.

Please make sure that you discuss the use of any topical solution in the study with your doctor or related field specialist before applying.

Please Donate for funding the larger sample size study to improve the validation of this discovery                             



Please print and complete the survey below and email me at kylenorton@live.ca 
World wide survey for Dehydroascorbic acid (DHA)(vitamin C oxidized) after 12

weeks of usage

1. Age group:

please circle (age group of 20 - 30, 31 - 40, 41 - 50, 51 -

60, 60 and up)

2. Ethnicity:

please circle (Caucasian, African, Asian, Other)

3. Gender:

please circle (Male or Female)

4. Concentrations usage

: please circle (2%, 3% or 4%, other

concentration)

5. Type of skin: (Please write)

6. Effectiveness range from 0 – 4: (0 - Not applicable, 1 - Minimum, 2

- Mild, 3 - Moderate, 4 - excellent effect).



Skin roughness and Scaliness: please circle

(0 - Not applicable, 1 - Minimum, 2 - Mild, 3 - Moderate, 4 - excellent effect).

Wrinkles: please circle (0 - Not applicable, 1 - Minimum, 2 – Mild, 3 -

Moderate, 4 - excellent effect).

Skin elasticity and firming: please circle

(0 - Not applicable, 1 - Minimum, 2 - Mild, 3 - Moderate, 4 - excellent effect).

Liver and aging spots: please circle

(0 - Not applicable, 1 - Minimum, 2 -Mild, 3 - Moderate, 4 - excellent effect).

Pigmentation (including photopigments): please circle

(0 - Not applicable, 1 - Minimum, 2 - Mild, 3 - Moderate, 4 - excellent effect).

Acne and Acne scars: please circle

(0 - Not applicable, 1 - Minimum, 2 - Mild, 3 - Moderate, 4 - excellent effect).

Back and White heads: please circle

(0 - Not applicable, 1 - Minimum, 2 -Mild, 3 - Moderate, 4 - excellent effect).

Fine Lines: please circle

(0 - Not applicable, 1 - Minimum, 2 - Mild, 3 - Moderate, 4 - excellent effect).

Please complete and send the survey to



References
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(c) Topical Dehydroascorbic Acid (Oxidized Vitamin C) Permeates Stratum Corneum More Rapidly Than Ascorbic Acid by Douglas Q Kitt
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(62) Green tea consumption is associated with lower psychological distress in a general population: the Ohsaki Cohort 2006 study by Atsushi Hozawa, Shinichi Kuriyama, Naoki Nakaya, Kaori Ohmori-Matsuda, Masako Kakizaki, Toshimasa Sone, Masato Nagai,Yumi Sugawara, Akemi Nitta, Yasutake Tomata, Kaijun Niu, and  Ichiro TsujiStudy(The American Journal of Nutrition)
(63)Green tea consumption is associated with depressive symptoms in the elderly. by Niu K1, Hozawa A, Kuriyama S, Ebihara S, Guo H, Nakaya N, Ohmori-Matsuda K, Takahashi H, Masamune Y, Asada M, Sasaki S, Arai H, Awata S, Nagatomi R, Tsuji I.(PubMed)
(64) Herbal Green tea (popular herb)
(65) Combined topical application of lotus and green tea improves facial skin surface parameters by Mahmood T1, Akhtar N.(PubMed)
(66) Tannase-converted green tea catechins and their anti-wrinkle activity in humans by Hong YH1, Jung EY, Shin KS, Yu KW, Chang UJ, Suh HJ.(PubMed)
(67) The use of green tea extract in cosmetic formulations: not only an antioxidant active ingredient by Gianeti MD1, Mercurio DG, Campos PM.(PubMed)
(68) Green tea polyphenols provide photoprotection, increase microcirculation, and modulate skin properties of women by Heinrich U1, Moore CE, De Spirt S, Tronnier H, Stahl W.(PubMed)
(69) Protective effects of (-)-epigallocatechin-3-gallate on UVA- and UVB-induced skin damage by Kim J1, Hwang JS, Cho YK, Han Y, Jeon YJ, Yang KH.(PubMed)
(70) UV-induced NF-kappaB activation and expression of IL-6 is attenuated by (-)-epigallocatechin-3-gallate in cultured human keratinocytes in vitro by Xia J1, Song X, Bi Z, Chu W, Wan Y.(PubMed)
(71)A comparison of the effects of topical green tea and lotus on facial sebum control in healthy humans.

Mahmood T1, Akhtar N, Moldovan C.(PubMed)
(72) Outcomes of 3% green tea emulsion on skin sebum production in male volunteers by Mahmood T1, Akhtar N, Khan BA, Khan HM, Saeed T.(PubMed)