AAVVIISS

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PRODUCT BROCHURE AVIS (AVOBENZONE USP)

INTRODUCTION

PRODUCT INFORMATION AVIS(AVOBENZONE USP) NEED OF SUNSCREENS Exposure to ultraviolet light, UVA or UVB, from sunlight accounts for 90% of the symptoms of premature skin aging such as wrinkles and skin cancers. The most important skin-care product available to prevent wrinkles and skin cancer is sunscreen.

Our challenge then, is to keep our skin protected from sun damage and excessive dryness during the long hours spent outside to train and compete

Protection needed for:

1. UVB (290 to 320 nm) - Burning of the skin and cancer risk

2. UVA (320 to 400 nm) - Causes skin to lose elasticity and promotes wrinkling, interferes with immune system

ABOUT SUN SCREENS

Sunscreens were initially developed to protect against UVB radiation. In the 1990s, compounds with UVA-absorbing ability also became available. Sunscreens are very effective at preventing erythema, the end point used in sun protection factor (SPF) determinations. Sunscreens are formulated using a number of different active compounds to increase the spectrum of absorption. The active sun screening agents are divided into two groups: organic and inorganic. The organic compounds protect the skin by absorbing UV radiation and inorganic compounds scatter UV radiation. EFFECTS OF UV RADIATION AND SUNSCREENS UV-radiation, especially UVB (called Burning Ray), produces dilatation of the blood vessels in the skin and, after prolonged exposure, an inflammation (erythema). Depending on the UV dose the skin melanocytes begin to produce melanin, a protein responsible for pigmentation. UVA (called Aging Ray) produces less erythema and pigmentation than UVB but damages the skin by penetrating deeply into the dermis able of producing premature aging, wrinkles, and tumors. Several studies have shown that the regular use of sunscreens is associated with a decrease in actinic keratoses (aged skin) and a decrease in a special type of skin cancer (squamous cell carcinomas).

PRODUCT INFORMATION AVIS(AVOBENZONE USP) SUN PROTECTION FACTOR (SPF) The sun protection factor is a measure of the ability of a sunscreen to protect against erythema, which is thus primarily a measure of UVB protection. The SPF is a ratio of the dose of UV radiation required to produce a minimal erythema 24 hours after exposure in sunscreen-protected skin to the dose required to produce the same degree of erythema in unprotected skin. In other words: the SPF number defines how long you can stay in the sun before getting burnt. If it normally takes you 20 minutes in the sun before you get burned, an SPF 15 product will let you stay 15 times longer in the sun: 20 min x 15 (SPF) = 300 min (5 hours). As global harmonization of SPF testing has progressed, the 2mg/cm2 application amount has become the worldwide standard for SPF testing. 2mg/cm2 is not an excessive amount of product; indeed, this application amount is required to adequately and evenly cover the test site for the SPF test. After the product is applied and rubbed in, there is no visible residue on the skin.

Even cloudy days pose a threat, as the suns UV rays can filter through overcast skies. Therefore, it is important to use a Broad Spectrum sunscreen all year round, to both protect against UVA and UVB rays and help retain youthful looking skin and firm facial contours.

PHOTOSTABILITY – SUN SCREENS

Labeled SPF Sunscreen active System

Percentage of total UV Absorbance Remaining after UV Exposure

5 Joules 10 Joules 50 Joules 8 OMC, OXY 100 100 100

15 OMC, PBSA, ZnO 100 100 100

15 OMC, PBSA 100 100 100

15 OMC, OS, OXY 100 100 100

15 TiO2, OMC 100 100 94

15 OMC, OS, TiO2 100 97 84

15 OMC, OCTO, OXY, TiO2 100 100 100

15 MA, OMC, TiO2 100 99 92

15 OMC, AVO, OXY 98 94 77

30 OMC, OXY, OS 100 100 100

30 OMCC, OCTO, MA, OS 100 100 100

30 OMC, OS, HS, OXY 100 100 100

PRODUCT INFORMATION AVIS(AVOBENZONE USP) Sunscreen Actives OMC = Octyl Methoxy Cinnamate OXY = Oxybenzone PBSA = Phenylbenzimidiazole Sulfonic Acid OS = Octyl Salicylate MA = Menthyl Anthranilate TiO2 = Titanium Dioxide OCTO = Octocrylene HS = Homosalate ZnO = Zinc Oxide AVO = Avobenzone THE MECHANISM OF ACTION SUNSCREENS The primary means of Ultra violet radiation by category I sunscreen actives utilized in commercial sunscreen products occurs through the mechanism of fluorescence or heat. This is true for both organic and inorganic molecules (although inorganic pigments can also reflect and scatter UV radiation). The degree of reflection and scattering of these pigments is strongly dependent on the particle size and shape. This is captured in the schematic diagram below. The organic sunscreen molecules are generally aromatic compounds conjugated with carbonyl groups. They absorb the high-energy ultraviolet photons, through electron resonance delocalization in the aromatic compounds, and are raised to a more energetic orbital state. The molecule quickly returns from the less stable excited state to the ground state, releasing the energy difference in longer (lower energy) wave lengths, either infrared (heat; >700nm), or visible (visible fluorescence; 400–700nm) radiation, thus satisfying energy conservation laws. At the ground state, the absorber is again available to absorb additional photons to repeat this cyclical process. This absorption of UV and emission of visible light/heat is the basis for how sunscreens function to protect human skin from deleterious effects of UV.

SCHEMATIC DIAGRAM

CH3CH3

O O

CH3

O

CH3

::

CH3CH3

O O

CH3

O

CH3

:

..: :

+

-

( )

( ) A third route that excited state molecules can potentially take involves the molecule undergoing photochemical reaction resulting in molecular change. The category I sunscreen that exhibits this behavior to a significant level is Avobenzone. This has been documented in published literature. Data on the photo stability of Avobenzone was reviewed by the Agency in 1996. FDA permitted marketing of Avobenzone in combination with specific Category I active Ingredients. Avobenzone has increased the range of UVA protections. It offers excellent absorption in the UVA II range, as well as in the UVA I region up to 380nm. It is used in combination with UVB absorbing chemicals such as the Cinnamates and salicylates. High-SPF products containing UVB absorbers, zinc oxide or titanium dioxide, Avobenzone, assure consumers of good photo protection from 290nm up to 400nm.

PRODUCT INFORMATION AVIS(AVOBENZONE USP) Chemical Structure:

OCH3

O O

C(H3C)3

Molecular Formula : C20 H22 O3 Molecular Weight : 310.4 Name of the product : AVIS Chemical Name : 4-t-butyl-4’-methoxy-dibenzoylmethane IUPAC Nomenclature : 1-(4-methoxyphenyl)-3-(4-tert-butylphenyl)- Propane-1, 3-dione INCI Name : Butyl Methoxy Dibenzoylmethane USAN : AVOBENZONE INN : AVOBENZONE CAS No : 70356-09-1 EINECS : 274-581-6

PRODUCT INFORMATION AVIS(AVOBENZONE USP) PRODUCT SPECIFICATION

S. No. TEST SPECIFICATION 01. Appearance White to pale yellow Powder 02. Solubility Insoluble in water. Soluble in Acetone 03. Melting Range (°C) Between 81°C and 86°C

04. Loss On Drying (Dry it in vacuum at 70° for 4 hours). ( % w/w)

Not more than 0.5% ( % w/w)

05. Specific Absorbance by UV (1% soln.in Ethanol;1cm cell at 357+ 2 nm)

1100 – 1180

06. Assay by GC (% w/w on the dried basis). Between 95.0 to 105.0 ( % w/w) APPLICATIONS UVA/UVB sunscreens, makeup creams, lotions, hand and face creams, medicated and moisturizing creams etc. Protects from UV-A spectrum, acting as an anti-aging agent. Avis provides broad-spectrum protection for skin and is widely used in sun care preparations. Used for the formulation of protective hair care preparations Used to Quench phototoxic skin reactions, initiated by weak photo initiators. As a photo antioxidant to protect products from photo-oxidation. In medicated skin care and protective skin tone preparations.

FINISHED PROUCT SPECIFICATIONS AVIS(AVOBENZONE USP) SPECIFICATIONS – AVIS(AVOBENZONE USP)

S.No Tests Specifications 1.0 Appearance Crystalline powder 2.0 Odor Characteristic weak aromatic odor 3.0 Color White to pale yellow 4.0 Solubility Soluble in Acetone &hot methanol, Insoluble in water.

5.0

Identification Test A Test B Solution:5µg per ml Medium: Alcohol

The IR absorption spectrum of the sample should be concordant with AVIS standard. Absorptivities at 360nm do not differ by more than 3.0%

6.0 Melting Range (in 0C) Between 810C and 860C

7.0 Loss on drying (Dry it in vacuum at 70 0C for 4 hours )

Not more than 0.5% w/w

8.0 Specific Absorbance (E 5µg per ml in alcohol, 1 cm cell, at 357 ± 2 nm)

1100 -1180

9.0 Chromatographic purity Individual Sum of the impurities

Not more than 3.0% Not more than 4.5%

10.0 Assay by GC Not less than 95.0% and not more than 105.0% w/w on the dried basis

Appearance Appearance is done at room temperature (25±50C) Apparatus: 4ml sample jar Reagents: Not required Procedure: Check the appearance of the sample The product should be white to pale yellow crystalline powder. Odor The odor of the sample is qualitatively determined to verify that the odor is characteristic for the product. Apparatus: 2ml sample jar Reagents: Odor standard Procedure: Pour the sample into a 2ml sample jar. Pour the standard sample into a 2ml sample jar. Open both jars and allow them to sit for 15 minutes at room temperature. Briefly smell the odor standard (less than 1 second) Briefly smell the sample (less than 1 second) Determine if the odor is characteristic of the product. Second individual must verify the odor test using the standard and the sample. Identification— A: Infrared Absorption Infrared Absorption — Four methods are indicated for the preparation of previously dried test specimens and Reference Standards for analysis. The substance under examination is mixed intimately with potassium bromide. The substance under examination is finely ground and dispersed in mineral oil. The substance under examination is suspended neat between suitable (for example, sodium chloride or potassium bromide) plates. The solution of designated concentration is prepared in the solvent specified in the individual monograph, and the solution is examined in 0.1mm cells unless a different cell path length is specified in the individual monograph. Record the spectra of the test specimen and the corresponding USP Reference Standard over the range from about 2.6 µm to 15 µm (3800 cm– 1 to 650 cm– 1) unless otherwise specified in the individual monograph. The IR absorption spectrum of the preparation of the test specimen, previously dried under conditions specified for the corresponding Reference Standard unless otherwise specified, or unless the Reference Standard is to be used without drying, exhibits maxima only at the same wavelengths as that of a similar preparation of the corresponding USP Reference Standard. Differences that may be observed in the spectra so obtained sometimes are attributed to the presence of polymorphs, which are not always acceptable . Unless otherwise directed in the individual monograph, therefore, continue as follows. If a difference appears in the IR spectra of the analyte

and the standard, dissolve equal portions of the test specimen and the Reference Standard in equal volumes of a suitable solvent, evaporate the solution to dryness in similar containers under identical conditions, and repeat the test on the residues B: Ultraviolet Absorption The test solution and a Standard solution are examined spectrophotometrically, in 1-cm cells, over the spectral range from 200 to 400 nm unless otherwise specified in the individual monograph. Dissolve a portion of the substance under examination in the designated Medium to obtain a test solution having the concentration specified in the monograph for Solution. Similarly prepare a Standard solution containing the corresponding USP Reference Standard. Record and compare the spectra concomitantly obtained for the test solution and the Standard solution. Calculate absorptivities and/or absorbance ratios where these criteria are included in an individual monograph. Unless otherwise specified, absorbances indicated for these calculations are those measured at the maximum absorbance at about the wavelength specified in the individual monograph. Where the absorbance is to be measured at about the specified wavelength other than that of maximum absorbance, the abbreviations (min) and (sh) are used to indicate a minimum and shoulder, respectively, in an absorption spectrum. The requirements are met if the UV absorption spectra of the test solution and the Standard solution exhibit maxima and minima at the same wavelengths and absorptivities and/or absorbance ratios are within specified limits. Solution: 5 µg per mL. Medium: alcohol. Absorptivities at 360 nm do not differ by more than 3.0%. C . Melting range : between 81° and 86°. Reduce the substance under test to a very fine powder, and, unless otherwise directed, render it anhydrous when it contains water of hydration by drying it at the temperature specified in the monograph, or, when the substance contains no water of hydration, dry it over a suitable desiccant for not less than 16 hours. Charge a capillary glass tube, one end of which is sealed, with a sufficient amount of the dry powder to form a column in the bottom of the tube 2.5 to 3.5 mm high when packed down as closely as possible by moderate tapping on a solid surface. Heat the bath until the temperature is about 30° below the expected melting point. Remove the thermometer, and quickly attach the capillary tube to the thermometer by wetting both with a drop of the liquid of the bath or otherwise, and adjust its height so that the material in the capillary is level with the thermometer bulb. Replace the thermometer, and continue the heating, with constant stirring, sufficiently to cause the temperature to rise at a rate of about 3° per minute. When the temperature is about 3° below the lower limit of the expected melting range, reduce the heating so that the temperature rises at a rate of about 1° to 2° per minute. Continue heating until melting is complete. The temperature at which the column of the substance under test is observed to collapse definitely against the side of the tube at any point is defined as the beginning of melting, and the temperature at which the test substance becomes liquid throughout is defined as the end of melting or the “melting point.” The two temperatures fall within the limits of the melting range.

D. Loss on drying : Dry it in vacuum at 70° for 4 hours: it loses not more than 0.5% of its weight. The procedure set forth in this chapter determines the amount of volatile matter of any kind that is driven off under the conditions specified. Mix and accurately weigh the substance to be tested, and, unless otherwise directed in the individual monograph, conduct the determination on 1 to 2 g. If the test specimen is in the form of large crystals, reduce the particle size to about 2 mm by quickly crushing. Tare a glass-stoppered, shallow weighing bottle that has been dried for 30 minutes under the same conditions to be employed in the determination. Put the test specimen in the bottle, replace the cover, and accurately weigh the bottle and the contents. By gentle, sidewise shaking, distribute the test specimen as evenly as practicable to a depth of about 5 mm generally, and not more than 10 mm in the case of bulky materials. Place the loaded bottle in the drying chamber, removing the stopper and leaving it also in the chamber. Dry the test specimen at the temperature and for the time specified in the monograph Upon opening the chamber, close the bottle promptly, and allow it to come to room temperature in a desiccator before weighing. If the substance melts at a lower temperature than that specified for the determination of Loss on drying, maintain the bottle with its contents for 1 to 2 hours at a temperature 5° to 10° below the melting temperature, then dry at the specified temperature. Where the specimen under test is Capsules, use a portion of the mixed contents of not less than 4 capsules. Where the specimen under test is Tablets, use powder from not less than 4 tablets ground to a fine powder. Where the individual monograph directs that loss on drying be determined by thermo gravimetric analysis, a sensitive electrobalance is to be used. Where drying in vacuum over a desiccant is directed in the individual monograph, a vacuum desiccator or a vacuum drying pistol, or other suitable vacuum drying apparatus, is to be used. Where drying in a desiccator is specified, exercise particular care to ensure that the desiccant is kept fully effective by frequent replacement. Where drying in a capillary-stoppered bottle in vacuum is directed in the individual monograph, use a bottle or tube fitted with a stopper having a 225 ± 25 µm diameter capillary, and maintain the heating chamber at a pressure of 5 mm or less of mercury. At the end of the heating period, admit dry air to the heating chamber, remove the bottle, and with the capillary stopper still in place allow it to cool in a desiccator before weighing. E. Chromatographic purity Test solution— Proceed as directed for Assay preparation in the Assay. Chromatographic system Proceed as directed in the Assay. Procedure— Inject a volume (about 1 µL) of Test solution into the chromatograph, record the chromatogram, and measure the peak responses. Calculate the percentage of each impurity in the portion of Avobenzone taken by the formula: 100(r I / r S), in which r I is the response of each individual peak, other than the avobenzone peak, in the chromatogram of the Test solution, and r S is the sum of the responses of all of the peaks in the chromatogram of the

Test solution: not more than 3.0% of any individual impurity is found, and the sum of all of the impurities is not more than 4.5%. F. Assay Standard preparation— Dilute an accurately measured quantity of USP Avobenzone RS in acetone, and dilute quantitatively, and stepwise if necessary, with acetone to obtain a solution having a known concentration of about 50 mg per mL. Assay preparation— Transfer about 500 mg of Avobenzone, accurately weighed, to a 10-mL volumetric flask, dilute with acetone to volume, and mix. Chromatographic system The gas chromatograph is equipped with a flame-ionization detector and a 0.32-mm × 25-m fused silica capillary column coated with phase G1. The column temperature is maintained at about 200° until the time of injection, then increased at a rate of 4° per minute to 280°. The injector port is maintained at 200°, and the detector is maintained at about 280°. Helium is used as the carrier gas. Procedure— Separately inject equal volumes (about 1 µL) of the Standard preparation and the Assay preparation into the chromatograph, record the chromatograms, and measure the responses for the major peaks. Calculate the quantity, in mg, of C 20H 22O 3 in the portion of Avobenzone taken by the formula: 10C(r U / r S), in which C is the concentration, in mg per mL, of USP Avobenzone RS in the Standard preparation; and r U and r S are the peak responses obtained from the Assay preparation and the Standard preparation, respectively.

EFFICACY STUDIES

Evaluation of sunscreen efficacy is most relevant when measured on the surface it is meant to protect, namely on human skin in vivo. Application of any material to the surface of the skin alters its optical properties. Diffuse reflectance spectroscopy (DRS) is a non-invasive technique to measure changes in the optical properties of the skin decoupled from its biological responses following sunscreen application. METHODS: This study compared measurements of UVA efficacy of oxybenzone and avobenzone at different concentrations (0-5%) using DRS, human phototest and an in vitro technique. Twenty subjects were enrolled for each product measured by DRS and 10 different subjects were enrolled for each product measured by human phototest. Six areas 5 cm x 10 cm were outlined on each subject's back. DRS measurements were performed on four subsites within each area before and 20 min after sunscreen application. UVA efficacy for each concentration of product was calculated from the measured transmission spectrum of a given product convoluted with the spectrum of a Xenon light source adequately filtered to obtain the UVA spectrum from 320 to 400 nm and the erythema action spectrum. Phototesting was performed using the same light source and persistent pigment darkening as the biological endpoint. Measurements were made with sunscreen coverage of 2 mg/cm2. In vitro measurements were performed using an Optometrics instrument. RESULTS: All three techniques showed a linear response between calculated UVA efficacy and product concentration. CONCLUSIONS: This study showed that DRS is a rapid and reproducible method to calculate UVA efficacy of sunscreen materials and that its results correlate closely with those obtained by human phototesting. REFERENCES

1. Lowe, N., Shaath, N. The Chemishy of Szcnscreens, in Summ~. 1 >eveloDment E&don. and Regulatorv 1990. Pp. 21 l-233. Armects.Marcel Del&r, Inc. New York 2. Shaath, N. on the theory of dt.rm~ok~ abso?pion by sunscreen chemic&.J. &c. CO= Cfiem. 82: 193-207, 1987. 3. Morliere, P. et al. A sh@ of the photochemical behavior of sunscreens. J. Photochem 30:2 15-277,1985. 4. D&la&e, Lang, G. PhotostiiIiq assessment of sunscreens. Benqylidene camphor and dibetuoylmethone derivatives. Int J. Cosm Sci. 1053-62, 1988. 5. West&t W., Kammeijer, J. Suggestion for Photostable Sunscreens. cosmetic Dermatology. 1: 301-3 11,1986.

6. Kammeya A et al. The spectraI stub&y of several sunscreening agents on stratum corneum sheetr. Int J.Cosm. Sci. 9:125-136. 1987. 7. Stenberg, C. et al. Stability of PBA ajkr Uvirradiation in viva and in vitro. Photcdetmatology 4: 201-204.1987. 8. Wellman Laboratories of Photomedicine, Massachusetts General Hospital, Boston, MA 02114, USA.

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