PhytoHyaluronic Acid

Transcription

1 PhytoHyaluronic Acid Chemical Nature - PhytoHyaluronic Acid is a non-microbially derived, water-soluble polymer composed of an α-(1-3)-mannose polysaccharide backbone with Fucose, β- Mannose, 3β-Glucuronic Acid, double-xylose, peptide and nucleic acid subunits and containing traces of Zinc, Copper, Iron. It is available in three molecular weight ranges: ~ 5, Da, ~ 8, Da and > 1,2, Da. Moisture Retention - PhytoHyaluronic Acid is an extraordinary humectant. Table 1 illustrates the in vitro moisture retention of three different humectants: PhytoHyaluronic Acid (PHA),.5%; Hyaluronic Acid (HA),.5%; Glycerin,.5%; and a control (distilled water). Table 1. Comparative Dehydration of Three Humectants at 25. C and a Constant Relative Humidity of 34%. Time PHA HA Glycerin (hr) ± ±.2** 6.1 ± ± ±.4** 13.2 ± ± ±.7** 29.7 ±.5* ± ± 1.** 41. ±.5* ± ± 1.2** 63.9 ± 1.3* *P <.5, **P <.1 These tabulated results, represented graphically in Figure 1, reveal that PhytoHyaluronic Acid is a superior humectant to both HA and glycerol. 8 6 Figure 1 PHA HA Glycerin Moisture Loss, % Time (hrs) Wilshire Technologies -2- TEL:

2 Skin Conductance Measurements - In vivo water sorption-desorption studies offer a window into humectant efficacy. In studies performed using the Skicon-2 skin conductivity apparatus, three determinations were carried out under standardized conditions [see: Tagami, et al., J.Investigative Dermatology, 78, 425 (1982)]. The first determination measured the pre-hydration level of the skin, an important indicator of the base-line hydration state of an individual s stratum corneum. A second series of tests were performed to determine the integrated skin conductance, a parameter that indicates the moisture-holding capacity of an individual s stratum corneum. The higher the value of this parameter. the healthier the skin. Rough, dry skin has a low skin conductance integration value. A final test was performed to measure the test subjects skin desorption rate constant which is an indicator of skin barrier function. This study relies on the relationship expressed in Eq. 1, which approximates the time dependence of water desorption from a defined area of skin surface. Where and W is the measured conductance k is the desorption rate constant t is the measured time W = W e -kt (Eq. 1) bviously, the smaller the value of k becomes, the more the skin-barrier function is improved. Using the test formulation defined in Table 2, a study of these three parameters Table 2. Reagent Placebo Reference Test PhytoHyaluronic - -.2% Sodium Hyaluronate -.2% - Phenoxyethanol.2%.2%.2% Methylparaben.1%.1%.1% Water to 1% to 1% to 1% The results are summarized in Figure 2. Wilshire Technologies -3- TEL:

3 Figure 2. Pre-hydration Levels as Measured on the Forearm of a Healthy Adult Human Placebo Lotion Sodium Hyaluronate - containing Lotion 8 8 Conductance, μs 6 4 Conductance, μs Time, sec 5 15 Time, sec PhytoHyaluronic Acid-containing Lotion 8 Conductance, μs Legend 4 weeks 2 weeks Initially 5 15 Time, sec Wilshire Technologies -4- TEL:

4 A summary of the percent change observed for each of the three in vivo water sorptiondesorption studies is summarized in Figure 3.. The superior moisturizing properties of PhytoHyaluronic Acid are evident. Figure 3 Pre-hydration Level Integrated Skin Conductance % Change bserved % Change bserved 2 4 Weeks 2 4 Weeks 13 % Change bserved Legend Placebo Sodium Hyaluronate PhytoHyaluronic Acid 2 4 Weeks Wilshire Technologies -5- TEL:

5 Anti-aging Properties of PhytoHyaluronic Acid - The role of free radicals, in particular the biological occurrence and destructive nature of hydroxyl and hydroperoxy radicals, in aging are well-documented. As the plot below demonstrates, PhytoHyaluronic Acid acts as an efficient scavenger of these potent oxidants, affording an in vitro reduction of 5% at.5% and a 7% reduction at 1% (Figure 4). Normalized Free Radical Conc Figure 4. Free-Radical Scavenging by PhytoHyaluronic Acid Source: Fenton Reagent, H 2 2 H. + H PhytoHyaluronic Acid, wt% The biological prevalence of the superoxide radical anion, 2, and it role as a precursor to hydroxyl and hydroperoxy radicals in biological membranes is generally acknowledged as is the role that the SD enzyme plays in mediating the level of this destructive agent. Figure 5 demonstrates that PhytoHyaluronic Acid is capable of enhancing the in vivo performance of SD in both keratinocytes and fibroblasts. Thus, when added to the culture media nurturing rat skin keratinocyte or fibroblast cells, the SD activity in both cells types increased after cultivating for 24 hrs at 37 C in 5% C2. Wilshire Technologies -6- TEL:

6 Figure 5. PhytoHyaluronic Acid Enhancement of SD Activity in cultured Skin Cells SD Activity, units/ml PhytoHyaluronic Acid, wt % Peroxidation of cellular lipids is known contributor to skin aging. In an experiment analogous to that described above, the incorporation of PhytoHyaluronic Acid into the culture media nurturing, respectively, rat skin keratinocyte and fibroblast cells, the level of peroxidized lipids, as judged by MDA production (as determined by TBA methodology), was suppressed (Figure 6). Wilshire Technologies -7- TEL:

7 Figure 6. PhytoHyaluronic Acid Suppression of Lipid Peroxidation in cultured Skin Cells MDA, mmol/ml PhytoHyaluronic Acid, wt % MDA = Malondialdehyde; TBA = Thiobarbituric Acid Wilshire Technologies -8- TEL:

8 Performance Stability A key consideration in the performance of any active cosmetic ingredient is the stability under working conditions. Inasmuch as viscosity is an intrinsic, fundamental property PhytoHyaluronic Acid in solution, the influence of temperature and ph on this parameter is a minimal requirement for assessing its performance stability. Figure 7a illustrates the influence that temperature has on the viscosity of a 1% aqueous solution of PhytoHyaluronic Acid, while Figure 7b depicts the influence of ph on the.5% solution over a temperature range of 25 ºC. Viscosity, m Pa s T, ºC Viscosity, m Pa s ºC, t = 25 ºC, t = 1 mo. 4 ºC, t = 1 mo. 5 ºC, t = 1 mo ph Figure 7a Figure 7b Figure 7. The influence of temperature and ph on the viscosity of a 1% aq. solution of PhytoHyaluronic Acid. Haake, Roto-Visco RV-1cone-plate viscometer These data reveal (1) that temperature has small, reversible effect on the solution viscosity of PhytoHyaluronic Acid and (2) that PhytoHyaluronic Acid has a broad ph stability (ph >3) for extended periods of time (>1 mo). Similar studies performed to determine the effect of mono- and divalent salts -- specifically, NaCl, MgCl2 and (PhS3)2Zn -- on the solution viscosity of PhytoHyaluronic Acid showed that concentrations of these salts as high as 1.5% by weight have no effect on the viscosity of 1% aq. solution of PhytoHyaluronic Acid. Finally, in studies performed to test the shelf-life stability of aqueous of PhytoHyaluronic Acid solutions, samples of 1% aq. solution containing an appropriate preservative (Germall) were placed in an environmental chamber and maintained there at a constant temperature 6 ºC for 1 month. Samples were periodically removed and subjected to centrifugation (4 rpm) for 15 min. At no time was any stratification or residue observed and all samples maintained there original clarity and color. Microbiological testing revealed a total bacteria count that was unchanged. Wilshire Technologies -9- TEL:

9 Formulation and Comparative Sensory Evaluation - What follows is a series of typical formulations and the subsequent results of blind sensory evaluations by a panel of cosmetologists. Moisturizing Cream Table 3. Comparison of two comparably formulated Moisturizing Creams containing, respectively, the active ingredient.1% PhytoHyaluronic Acid and.2% Hyaluronic Acid. No. Ingredient Wt % Wt % 1 PhytoHyaluronic Acid Sodium Hyaluronate Simmondsia Chinensis (Jojoba) Seed il Squalane Behenyl Alcohol Stearyl Alcohol Hydrogenated Palm il Glyceryl Stearate Propylene Glycol Stearate Peg-6 Glyceryl Isostearate Ethylparaben Dimethicone Butylene Glycol Glycerin Phenoxyethanol Methylparaben Water to 1 to 1.1% PhytoHyaluronic Acid.2 % Sodium Hyaluronate Moistness Suppleness Instructions Step 1: Prepare Part A by mixing components 3-11, heating to 7 ºC and blending until uniform. Step 2: Prepare Part B by mixing components 1 and 12-16, heating the mixture to 7 ºC and blending until uniform. Step 3: Add Part-B to Part-A and homogenizing for 3 minutes; allow to cool. Wilshire Technologies TEL:

10 Lotion Table 4. Comparison of two comparably formulated Lotion containing, respectively, the active ingredient.2% PhytoHyaluronic Acid and.2% Hyaluronic Acid. No. Ingredient Wt % Wt % 1 PhytoHyaluronic Acid Sodium Hyaluronate Citric Acid Sodium Citrate Phenoxyethanol Methylparaben Water to 1 to 1.1% PhytoHyaluronic Acid.2 % Sodium Hyaluronate Moistness Suppleness Smoothness Sticky Instructions Step 1: Prepare Part A by mixing components 4 & 5 together with 5% of component 6 and heating to 7 ºC. Blend until uniform. Step 2: Prepare Part B by mixing component 1 with the remaining 5% of component 6 and blending until uniform. Step 3: Combine Part B, Part A and components 2 and 3 and homogenize until uniform. Hair Mist - Adds moist, airy body to hair No. Ingredient Wt % 1 PhytoHyaluronic Acid.2 2 Butylene Glycol 1. 3 Glycerin 3. 4 Sodium PCA.3 5 Alcohol 5. 6 Sodium Citrate.1 7 Methylparaben.1 8 Water to 1 Heat components 1-8 at 8 C and stir until uniform; allow to cool. Wilshire Technologies TEL: Formulating Notes (1) PhytoHyaluronic Acid should be thoroughly admixed with water before blending with non-aqueous phase. (2) Because of its rich nutrient nature, PhytoHyaluronic Acid should always be formulated with an appropriate preservative, e.g. Germall, etc.

11 Jump to Website Request Quote Hyaluronic Acid, a plant-derived, non-heteroatom containing polysaccharide, (PhytoHyaluronic ; Tremella fuciformis sporocarp) CAS INCI Name: Tremella fuciformis sporocarp polysaccharide CFTA Name: Tremella Hyaluronic Acid Composition: Structure: An α-(1-3)-mannose Polysaccharide backbone with Fucose, 3β-Glucuronic Acid, double-xylose, peptide and nucleic acid subunits. H H H H H * H H H H H H H H H C 2 H H H H H H C 2 H CH 2 H H H H H H H H H H H H H H H H H C 2 H H H H H H H H H H H H * n ~ 2-4 Mol. Formula N/A Mol. Wt. > 1. x1 6 Daltons Source Tremella fuciformis sporocarp Mnft Date Lot No. Re-test - cont d- Wilshire Technologies TEL:

12 - cont d- Test Specification Results Identity (UV-Vis) Appearance rganoleptic Glucuronic Acid Taste Tasteless dor dorless Assay Total saccharides (Phenol-Vitriol test) > 8% Glucuronic Acid (Sulfuric Acid - Carbazole or 13-3% - m-phenylphenol test) Nitrogen (Kjeldahl) <.5% LD (15 C, 4 h) < 1% ph (.5% in water) Absolute Viscosity (.5% aq. soln.) Particle size (ASTM) Carbohydrates (pentoses, hexoses) suffer dehydration in strong acid resulting, respectively, in formation of Furfuraldehyde and 5-Hydroxyfurfuraldehyde, whose subsequent condensation with any of a variety of phenols produces a highly colored phenol-furfural intermediate with an intense absorption between 4-6 nm. Sulfuric Acid Carbazole test results in an intense absorption at 535 nm - or - Sulfuric Acid m-phenylphenol test results in an intense absorption 52 nm White to pale-yellow powder [glass electrode] Pa s [Brookfield viscometer, spindle # 63, C] 8 mesh Residue on Ignition, USP <281> < 1% Mol. Wt. (a) Ubbelohde viscometer (b) GPC/SEC (Light scattering /RI detection) (c) MALDI-TF > 1. x 1 6 Da (Viscosity-Average MW) > 2. x 1 6 Da (Weight-Average MW) > 1.3 x 1 6 Da (parent oligomer) Transparency (.5% aq. 4 nm) > 9% - cont d- Wilshire Technologies TEL:

13 - cont d- Test Specification Results Residual Solvent EtH Microbiological Specification Total Aerobic, USP <61> Coliform Staphylococcus aureus Salmonella Shigella Streptococcus hemolyticus Mold, USP <61> Aflatoxins (GB/T ) B 1 B 2 G 1 G 2 Heavy metals, USP <231> As Pb Hg Cd Cr Ni Sb Metals, USP <851> Na K Mg Zn Nutritional Content Energy Protein Fat Carbohydrates Storage: Maintain in cool, dry containment. < 1, ppm < 1 cfu/g < 3 MPN/1 g ND ND ND ND < 1 cfu/g <.5 μg/kg <.125 μg/kg <.5 μg/kg <.125 μg/kg < 1.5 ppm < 2 ppm <.6 ppm <.3 ppm <.2 ppm <.1 ppm <.1 ppm < 1 ppm < 5 ppm < 1 ppm < 1 ppm ~ 363 Kcal/1 g ~.76 g/1 g ~.24 g/1 g ~ 9/1 g Non-GM: This product is derived from a non-gm vegetal source as designated under EP regulations 1829/23 and 183/23. TSE/BSE: The source of this product is pure Tremella fuciformis sporocarp. During production, storage and transport, all contact with any material of animal origin was excluded. Therefore, the requirements set forth in of the EP Comm. Directive 1999/82/EEC, CPMP/BWP/123/98, and EP 1/28: 528 do not apply. Wilshire Technologies TEL: