UVA Protection Factor Research Articles

Structural elucidation, in vitro antioxidant and photoprotective capacities of a purified polyphenolic-enriched fraction from a saltmarsh plant

In temperate saltmarshes, halophytic plants have to daily protect their internal tissues against sunlight and UV rays. Consequently, they develop adaptive responses such as the synthesis of secondary metabolites, including polyphenols. The present study focused on the biological activities of fractions enriched in polyphenols from Salicornia ramosissima. Three different extracts were obtained by purification processes to concentrate polyphenols: a crude hydroalcoholic extract, and two purified fractions: an ethyl acetate fraction (EAF) and an aqueous fraction. Phenolic and flavonoid contents, antioxidant (DPPH radical-scavenging activity, reducing activity, β-carotene linoleic acid system and the ORAC method) and sunscreen properties (Sun Protection Factor and UVA-Protection Factor) were assessed by in vitro tests. The purification process was effective in increasing phenolic and flavonoid contents as well as antioxidant and sunscreen capacities of the EAF. The EAF appeared to be a broad spectrum UV absorber. The chemical structure of 10 EAF polyphenols was elucidated using 2D NMR and mass spectrometry spectra. Furthermore, a correlation was observed between phenolic composition and biological activity. These findings are encouraging for the future use of S. ramosissima as a potential source of antioxidant and photoprotectant molecules for industrial applications.

A broad spectrum high-SPF photostable sunscreen with a high UVA-PF can protect against cellular damage at high UV exposure doses.

Advances in sunscreen technologies have yielded broad spectrum sunscreens at high-sun protection factor (SPF) and ultraviolet A protection factor (UVA-PF) levels that are photostable and powerful in protecting skin from erythema. Questions arise whether these sunscreens protect proportionally against cellular skin damage caused by high ultraviolet exposures. The objective of this study is to evaluate if high-SPF sunscreen can protect skin at a cellular level under UV exposure doses [>50 minimal erythema dose (MED)] similarly to the SPF value. Sunburn cells, Langerhans cells, thymine dimers, protein 53 (p53), and matrix metalloproteinase (MMP)-1 and MMP-9 endpoints were evaluated in biopsies from 12 subjects following four treatments: unprotected exposed to 0, 1 and 3 MED and sunscreen (SPF 55) protected exposed to 55 MED of UV radiation. All the markers showed significantly more damage for the 3 MED-untreated sites when compared with non-irradiated control, and majority of the markers showed marked damage following unprotected 1 MED exposure. After 55 MEDs, sunscreen-protected sites showed significantly less p53 and MMP-9 (keratinocyte) staining than the 1 MED-exposed unprotected sites, while all the other biomarkers in sunscreen protected sites showed no statistical differences from 1 MED-exposed unprotected sites. A high-SPF photostable sunscreen with high UVA-PF can provide proportionately high protection against multiple cellular damage markers.

New noninvasive approach assessing in vivo sun protection factor (SPF) using diffuse reflectance spectroscopy (DRS) and in vitro transmission

In the past 56 years, many different in vitro methodologies have been developed and published to assess the sun protection factor (SPF) of products, but there is no method that has 1:1 correlation with in vivo measurements. Spectroscopic techniques have been used to noninvasively assess the UVA protection factor with good correlation to in vivo UVA-PF methodologies. To assess the SPF of sunscreen product by diffuse reflectance spectroscopy (DRS) technique, it is necessary to also determine the absorbance spectrum of the test material in the UVB portion of the spectrum (290-320 nm). However, because of the high absorbance characteristics of the stratum corneum and epidermis, the human skin does not remit enough UVB radiation to be used to measure the absorption spectrum of the applied product on skin. In this work, we present a new method combining the evaluation of the absolute UVA absorption spectrum, as measured by DRS with the spectral absorbance 'shape' of the UVB absorbance of the test material as determined with current in vitro thin film spectroscopy. The measurement of the in vivo UVA absorption spectrum involves the assessment of the remitted intensity of monochromatic UVA radiation (320-400 nm) before and after a sunscreen product was applied on skin using a spectrofluorimeter Fluorolog 3, FL3-22 (Yvon Horiba, Edison, NJ, USA). The probe geometry assures that light scattering products as well as colored products may be correctly assessed. This methodology has been extensively tested, validated, and reported in the literature. The in vitro absorption spectrum of the sunscreen samples and polyvinyl chloride (PVC) films 'surrogate' sunscreen standards were measured using Labsphere® UV-2000S (Labsphere, North Sutton, NH, USA). Sunscreens samples were tested using PMMA Helioplates (Helioscience, Marseille, France) as substrates. The UVB absorbance spectrum (Labsphere) is 'attached' to the UVA absorbance spectrum (diffuse reflectance) with the UVB absorbance matched to the UVA absorbance at 340 nm to complete the full spectral absorbance from which an estimate the SPF of the product can be calculated. Seventeen test materials with known in vivo SPF values were tested. Two of the tested products were PVC sunscreen thin films with 10-15 micrometers thickness and were used to investigate the absorption spectrum of these films when applied on different reflectance surfaces. Similar to the human in vivo SPF test, the developed methodology suggests limiting the use on Fitzpatrick skin phototypes I to III. The correlation of this new method with in vivo clinical SPF values was 0.98 (r2) with a slope of 1.007. This new methodology provides a new approach to determine SPF values without the extensive UV irradiation procedures (and biological responses) currently used to establish sunscreen efficacy. Further work will be conducted to establish methods for evaluation of products that are not photostable.

SYNTHESIS AND EVALUATION OF OCTOCRYLENE-INSPIRED COMPOUNDS FOR UV-FILTER ACTIVITY

Octocrylene (2-ethylhexyl 2-cyano-3,3-diphenyl-2-propenoate) is present in several sunscreens and is known to work synergistically with UV filters. We prepared eight octocrylene-related compounds to test their photoprotective activities by measuring diffuse transmittance. The compounds had varied photoprotection profiles, with Sun Protection Factors (SPF) ranging from 1 to 5 and UVA Protection Factors (UVAPF) ranging from 1 to 8. Compounds 4, 5, and 7 showed the best protection against UVB sunrays, while compounds 5, 6, and 7 presented the best results for protection from UVA, so compound 7 had the most balanced protection overall. Results for compounds 4, 8, and 9 are reported for the first time in the literature.

Assessment of extracts of Helichrysum arenarium, Crataegus monogyna, Sambucus nigra in photoprotective UVA and UVB; photostability in cosmetic emulsions

The aim of our study was to investigate the photoprotective activity and photostability efficacy of sunscreen formulations containing Helichrysum arenarium, Sambucus nigra, Crataegus monogyna extracts and their combination. UV transmission of the emulsion films was performed by using diffuse transmittance measurements coupling to an integrating sphere. In vitro photoprotection and photostability efficacy were evaluated according to the following parameters: sun protection factor (SPF), UVA protection factor (PF-UVA), UVA/UVB ratio and critical wavelength (λc) before and after UV irradiation. The results obtained show that the formulations containing polyphenols fulfill the official requirements for sunscreen products due to their broad spectrum of UV protection combined with their high photostability and remarkable antioxidant properties. Therefore H. arenarium, S. nigra, C. monogyna extracts represent useful additives for cosmetic formulation.

Coencapsulation of Butyl‐Methoxydibenzoylmethane and Octocrylene into Lipid Nanocarriers: UV Performance, Photostability and in vitro Release

The coencapsulation of two UV filters, butyl-methoxydibenzoylmethane (BMDBM) and octocrylene (OCT), into lipid nanocarriers was explored to develop stable cosmetic formulations with broad-spectrum photoprotection and slow release properties. Different types of nanocarriers in various concentrations of the two UV filters were tested to find the combination with the best absorption and release properties. Solid lipid nanoparticles (SLNs) and nanostructured lipid carriers (NLCs) have been the two types of lipid nanocarriers used. The NLCs were based on either medium chain triglycerides (MCT) or squalene (Sq). The following physicochemical properties of the nanocarriers have been evaluated: particle size, morphology, zeta potential (ZP), entrapment efficiency, loading capacity, and thermal behavior. The nanocarriers have been formulated into creams containing low amounts of UV filters (2.5% BMDBM and 1% OCT). The best photoprotection results were obtained with the cream based on NLCs prepared with MCT, having a sun protection factor (SPF) of 17.2 and an erythemal UVA protection factor (EUVA-PF) of 50.8. The photostability of the encapsulated BMDBM filter was confirmed by subjecting the nanocarriers-based creams to in vitro irradiation. The prolonged UV-protection efficacy was coupled with a slow in vitro release of the synthetic UV filters, which followed the Higuchi release model.

Interoperability of 300 watt and 150 watt xenon arc solar simulators in sun protection factor and in UVA protection factor clinical testing

The purpose of these clinical trials was to determine if 300 W and 150 W xenon arc solar simulators (SSs) deliver the same sun protection factor (SPF) and UVA protection factor (PFA). First, the SPF of the P7 control standard and of the P2 control standard was determined, testing 20 subjects using the method described in the Food and Drug Administration (FDA) Final Monograph and using 150 W and 300 W SSs. In the second clinical trial, the PFA of the Japanese Cosmetic Industry Association (JCIA) control standard and of the P2 control standard was determined, testing 10 subjects using the method described in the JCIA Technical Bulletin and using 150 W and 300 W SSs. The SPF values for P7 control standard determined using the 150 W and 300 W SSs were 4.54 ± 0.35 and 4.61 ± 0.32, respectively. The SPF values for P2 control standard determined using the 150 W and 300 W SSs were 17.0 ± 0.9 and 16.7 ± 0.9, respectively. The resultant PFA values for JCIA control standard determined using the 150 W and 300 W SSs were 4.06 ± 0.70 and 4.06 ± 0.70, respectively. The resultant PFA values for P2 control standard determined using the 150 W and 300 W SSs were 3.28 ± 0.25 and 3.44 ± 0.39, respectively. As the values are essentially identical for SPF and for PFA, the 150 W and 300 W SSs can be used interchangeably for SPF and PFA determinations.

The revised COLIPA in vitro UVA method

A multicentred study derived from the COLIPA in vitro UVA method was performed to assess the influence of test conditions on UVA protection factor (UVAPF) values in terms of amplitude, reproducibility between laboratories and correlation with in vivo UVA results. Eight products with a range of in vivo UVAPF from three to 29 were used. Two different types of plates, namely high-roughness (5μm) and low-roughness (2μm) plates, were used with a different application rate for each (1.3mgcm(-2) and 0.75mgcm(-2) respectively). The UVR dose applied to both plate types followed the same principle as the original test (1.2J.cm(-2) ×UVAPF0). Strong, significant correlations between in vitro and in vivo UVAPF values were observed for both plate types (Pearson correlation>0.9, P≤0.01). The correlation and slope obtained with the low-roughness plates confirmed the previous results obtained by COLIPA. Across all laboratories, higher UVAPF values were obtained on the high-roughness plates (P<0.01). Reproducibility of UVAPF values between laboratories was comparable between the two plate roughness values (low roughness, COV=8%; high roughness, COV = 12%). Considering the in vitro/in vivo comparisons, a regression slope of 0.83 was observed for the low-roughness plates, in comparison with a value of 1.05 for the high-roughness plates. The accuracy of the method was improved, therefore, with the use of the high-roughness plates. With a constraint to recommend the use of only one plate type in the COLIPA UVA in vitro Test, the high-roughness plate was selected on an on-going basis to limit variability of results and to provide better accuracy with in vivo data.

Characterization and Evaluation of Poly(ε-caprolactone) Nanoparticles Containing 2-Ethylhexyl-p-Methoxycinnamate, Octocrylene, and Benzophenone-3 in Anti-Solar Preparations

Ultraviolet radiation can bring both harm and benefits to human health. Among those harms are erythemas, photosensitivity, photoaging, and the most worrying, skin cancer. Nanoencapsulation of sunscreen agents (SA) by using a biocompatible and biodegradable polymer such as poly(epsilon-caprolactone) (PCL) is advantageous as it increases the retention of UV absorbers in the skin, avoids systemic absorption, and consequently, improves water resistance and stability of the preparation. The aim of this work is to develop, characterize, and study the encapsulation of 3 different SA: 2-ethylhexyl-p-methoxycinnamate, benzophenone-3, and octocrylene in PCL nanoparticles (Nps). Nps were prepared by the solvent emulsification and evaporation method. The process yield was calculated, and the Nps were characterized in terms of size, polydispersity index (PI), morphology, zeta potential (ZP), encapsulation efficiency (EE) (%), and sunscreen agent content (SAC). The final formulations were submitted to the hen's egg test-chorioallantoic membrane (HET-CAM), chorioallantoic membrane-trypan blue staining (CAM-TBS), red blood cell (RBC), Draize tests, in vitro release, in vitro sun protection factor (SPF), UVA protection factor (PF-UVA), and photostability. All the Nps were in the nanometric scale. PI showed monodisperse systems. ZP became more negative as the Np were lyophilized and were added to the formulations. EE varied from 84 to 90%. The SAC went from 44 to 65 microg of sunscreen agents by milligram of Np. The process yield went from 60 to 76%. Nps were predominantly spherical and elliptical forms. The addition of Np diminished the release of the SA. The SPF increased with Np presence and helped to maintain the PF-UVA after irradiation. The HET-CAM assay evaluated the formulation as slightly irritant, CAM-TBS and RBC tests as non irritant, and the Draize test as moderately irritant.

Modulations of gene expression induced by daily ultraviolet light can be prevented by a broad spectrum sunscreen

Realistic non-zenithal solar ultraviolet (UV) exposure, obtained using standard ultraviolet daylight spectrum (DUVR), has deleterious impact on epidermal and dermal compartments of human skin. The present study was designed to assess gene expression in human reconstructed skin following exposure to DUVR and the protective effect of a broad spectrum sunscreen.Reconstructed skins were exposed to a realistic daily UV dose of 12J/cm2 DUVR in the presence of a sunscreen product (Sun(burn) Protection Factor (SPF)=13 and UVA protection factor UVAPF (PPD) 10.5) or its vehicle. Six hours post exposure, gene expression was investigated in fibroblasts (225 genes) and keratinocytes (244 genes) separately using quantitative PCR arrays.DUVR exposure led to significant modulation of 35 and 66 genes in fibroblasts and keratinocytes, respectively. These genes were involved in extracellular matrix homeostasis, oxidative stress response, cell growth, inflammation and epidermal differentiation. Sunscreen use significantly reduced DUVR-induced gene modulation. Hierarchical clustering showed that gene expression profiles in protected and DUVR-exposed samples were very close to those of unexposed samples. The number of DUVR-modulated genes was significantly decreased by tested sunscreen (zero and four modulated genes in fibroblasts and keratinocytes, respectively).Our results demonstrate that a broad-spectrum sunscreen product is highly effective in protecting reconstructed human skin against DUVR-induced changes in gene expression.

Lipid nanoparticles based on butyl-methoxydibenzoylmethane: in vitro UVA blocking effect

The aim of the present study was to obtain efficient lipid nanoparticles loaded with butyl-methoxydibenzoylmethane (BMDBM) in order to develop cosmetic formulations with enhanced UVA blocking effect. For this purpose, two adequate liquid lipids (medium chain triglycerides and squalene) have been used in combination with two solid lipids (cetyl palmitate and glyceryl stearate) in order to create appropriate nanostructured carriers with a disordered lipid network able to accommodate up to 1.5% BMDBM. The lipid nanoparticles (LNs) were characterized in terms of particle size, zeta potential, entrapment efficiency, loading capacity and in vitro UVA blocking effect. The efficiency of lipid nanoparticles in developing some cosmetic formulations has been evaluated by determining the in vitro erythemal UVA protection factor. In order to quantify the photoprotective effect, some selected cream formulations based on BMDBM-LNs and a conventional emulsion were exposed to photochemical UV irradiation at a low energy to simulate the solar energy during the midday. The results obtained demonstrated the high ability of cream formulations based on BMDBM-LNs to absorb more than 96% of UVA radiation. Moreover, the developed cosmetic formulations manifest an enhanced UVA blocking effect, the erythemal UVA protection factor being four times higher than those specific to conventional emulsions.

New Combination of Ultraviolet Absorbers in an Oily Emollient Increases Sunscreen Efficacy and Photostability

IntroductionIt is now recognized that to adequately protect skin from sun damage, sunscreens require a photostable combination of ultraviolet (UV) filters with a suitable level of UVA protection. The total amount of UV filters should be as low as possible to avoid adverse skin reactions, potential environmental impact, and to ensure acceptable texture for better application and usage.MethodsA synergistic combination of UV filters was selected to obtain a high sun protection factor (SPF) and UVA protection factor (UVA-PF). An oily vehicle was then added to the formula to improve the solubility and the photostability of the lipophilic UV filters.ResultsThe combination of filters, i.e., terephthalylidene dicamphor sulfonic acid (TDSA), bis-ethylhexyloxyphenol methoxyphenyl triazin (BEMT), and butyl methoxy dibenzoyl methane (BMDM), obtained an elevated SPF as well as a high UVA-PF. Isopropyl lauroyl sarcosinate (ILS), a derivative of a natural amino acid (sarcosine, also known as N-methylglycine) was introduced in this formulation in order to dissolve the oil-soluble UV absorbers and to photostabilize BMDM. The new sunscreen formulation obtained with this combination is photostable and contains a reduced amount of UV filters compared to other sunscreens with the same level of efficacy.ConclusionThis report described the steps resulting in the formulation of a new combination of UV filters in an oily emollient, which presents a high UVA-PF (UVA-PF = 38) and a SPF 50+, is photostable, and offers good protection against UV-induced biological damage.

SPEX SkinSkanによるサンスクリーン剤のUVA Protection Factorの検討

SPEX SkinSkanによるサンスクリーン剤のUVA Protection Factorの検討

Sunscreen products: What do they protect us from?

Whereas for fifty or so years acquiring a tan has been the trend for aesthetic reasons relating to current beauty criteria, the health authorities are now advocating vigilance in this area, prompted by the knowledge of the harmful effects of the sun, especially from its ultra-violet rays. In the European Union, sunscreen products are considered cosmetics and tests on their effectiveness can be performed in vivo or in vitro to determine four effectiveness indicators: the SPF (Sun Protection Factor), the PF-UVA (UVA Protection Factor), the SPF/PF-UVA ratio and the critical wave length. It is the erythemal SPF which is measured in the vast majority of cases: it can therefore be confirmed that sunscreen products protect us from sunburn under good conditions of use. We thought it would be interesting to calculate other indicators to assess protection against non-melanoma skin cancers (NMSC) and to quantify the effectiveness of the product against UVA1 or UVA2. To characterize the products tested, we have determined in vitro different SPF and PF-UVA values, by using not just the erythemal weighting factor but also the weighting factor relating to the non-melanocytic skin cancer (SPF cnm and PF-UVA cnm), by getting away from any weighting factor (SPF m and PF-UVA m) and lastly, by varying the integration limits to quantify the effectiveness of the tested product in the UVB (290–320 nm), UVA1 (340–400 nm) and UVA2 320–340 nm) fields. In this way, and using these new indicators, we have been able to qualify eleven commercial products—ten cosmetic products and one medical device. It can be interesting to take into account the non-melanocytic skin cancer protection in order to qualify the sunscreen products.

Immediate pigment darkening and persistent pigment darkening as means of measuring the ultraviolet A protection factor in vivo: a comparative study

Persistent pigment darkening (PPD) is a widely used in vivo method for measurement of ultraviolet (UV) A protection factor (UVAPF). However, with increased emphasis on UVA protection and sunscreen products with higher UVAPF gaining popularity, the immediate pigment darkening (IPD) method is drawing attention again. Furthermore, only about a quarter of the recommended quantity of sunscreen is used during daily activities. However, there is as yet no clearly defined relationship between the UVAPF and the amount of sunscreen applied. To analyse the differences between the IPD and PPD methods, and to establish a relationship between the quantity of sunscreen application and the UVAPF. Different doses of sunscreen were applied on the back of 15 healthy volunteers, and the UVAPF was measured using both the IPD and the PPD methods. Both methods proved to be effective for measuring the UVAPF. However, all the UVAPF values determined by the PPD method were lower than those determined by the IPD method. Additionally, an exponential relationship between the amount of sunscreen applied and the UVAPF was observed. The IPD method can also be used as an appropriate endpoint in the determination of UVA protection. It is time saving, and thus considerably lowers the risk of UV exposure, particularly when testing sunscreen products with higher UVAPF. We further state that in order to achieve the desired protective effect of the sunscreen, the quantity of application is also very important.

Characterization of the UVA Protection Provided by Avobenzone, Zinc Oxide, and Titanium Dioxide in Broad-Spectrum Sunscreen Products

Solar UV radiation (UVR) is composed of UVB (290-320 nm) and UVA (320-400 nm) wavelengths. Only two sunscreen active ingredients approved in the US, avobenzone (butylmethoxydibenzoylmethane) and zinc oxide (ZnO), provide true broad-spectrum protection against UVA wavelengths >360 nm. Although effective against shorter UVR wavelengths <360 nm, titanium dioxide (TiO(2)) is also often believed to confer broad-spectrum protection and is substituted for ZnO or avobenzone. To sustain its absorption capacity within a sunscreen film during UVR exposure, avobenzone needs to be formulated into sunscreen products using sound formulation strategies. To characterize the efficacy of avobenzone, ZnO, and TiO(2) in terms of their abilities to provide broad UVA protection and to demonstrate the effectiveness of the different formulation strategies used today to maintain the efficacy of avobenzone even during prolonged exposures to UVR. UVA efficacy was assessed by measuring absorbance profiles in vitro using Vitro Skin® (IMS Inc., Orange, CT, USA) as an inert substrate and by determining UVA protection factors (PFA) on human skin. The impact of avobenzone loss on sun protection factor (SPF) and PFA values was evaluated by serially reducing avobenzone concentrations in an otherwise photostable product. The photostabilizing influence of specific formulation ingredients was monitored by measuring the extent to which they prevented UVR-induced degradation of avobenzone, whereas photostability of commercial sunscreen products was quantified by measuring the percentage change in absorbance within the UVB and UVA spectral regions following irradiation of thin product films on inert substrates. Model formulations containing 3% avobenzone or 5% ZnO provided superior attenuation of UVA wavelengths >360 nm compared with formulas containing 5% TiO(2). Additionally, sunscreen products of similar SPF containing avobenzone or ZnO exhibited significantly higher PFA values than those containing TiO(2). The addition of photostabilized avobenzone or ZnO increased PFA values nearly 3-fold, whereas the addition of TiO(2) increased PFA values only modestly. Judicious selection of sunscreen actives alone or in combination with extra stabilizing agents maintained the photostability of avobenzone in formulations to deliver sustained broad-spectrum absorbance during 4 hours of exposure to UVR. Small losses (<20%) of avobenzone did not significantly reduce a product's protective effects as measured by SPF and PFA values on human skin. TiO(2) provided neither the same level of UVA attenuation nor the same degree of UVA protection on human skin as did products containing photostabilized avobenzone or ZnO. Hence, TiO(2) cannot be considered a substitute for avobenzone or ZnO in providing high levels of UVA protection to human skin. Use of proper formulation strategies can ensure that avobenzone losses are minimized to the extent that they have no impact on a product's ability to deliver sustained protection, even over periods of prolonged exposure to UVR.

Formulation of a Novel Oxybenzone-Loaded Nanostructured Lipid Carriers (NLCs)

The objective of the current study was to formulate oxybenzone into nanostructured lipid carriers (NLCs) to enhance its sunscreening efficacy and safety. NLCs of oxybenzone were prepared by the solvent diffusion method. A complete 2(3) factorial design was used for the evaluation of the prepared oxybenzone NLCs. The study design involves the investigation of the effect of three independent variables namely liquid lipid type (Miglyol 812 and oleic acid), liquid lipid concentration (15% and 30%), and oxybenzone concentration (5% and 10% with respect to total lipids) on the particle size (p.s.) , the entrapment efficiency (EE%) and the in vitro drug release after 8 h. The prepared NLCs were spherical in overall shape and were below 0.8 microm. Miglyol 812 and 30% liquid lipid were found to significantly decrease the p.s. and increase the EE% when compared to oleic acid and 15% liquid lipid. Increasing oxybenzone concentration increased significantly the p.s. but did not affect the EE%. NLCs prepared using Miglyol 812, 15% liquid lipid, and 10% oxybenzone showed slower drug release when compared to those prepared using oleic acid, 30% liquid lipid, and 5% oxybenzone, respectively. The candidate oxybenzone-loaded NLC dispersion was then formulated into gel. The incorporation of oxybenzone into NLCs greatly increased the in vitro sun protection factor and erythemal UVA protection factor of oxybenzone more than six- and eightfold, respectively, while providing the advantage of overcoming side effects of free oxybenzone as evidenced by very low irritation potential.

The COLIPA in vitro UVA method: a standard and reproducible measure of sunscreen UVA protection

There is a continuing need to measure and communicate reliably the UVA protection offered by commercial sunscreens. To that end, the COLIPA (European Cosmetics Trade Association) 'In Vitro Sun Protection Methods' group has developed a new in vitro method for measuring UVA protection in a standardized, reproducible manner. The method is based on in vitro UV substrate spectrophotometry and convolution of resulting absorbance data with the action spectrum for the in vivo Persistent Pigment Darkening (PPD) endpoint to provide an in vitro UVA protection factor (UVAPF) which is correlated with an in vivo measure. This method has been published as a COLIPA guideline, used currently in European geographies for testing and labelling sunscreen products. This article summarizes two 'ring' studies, involving eight separate testing laboratories, which both defined critical parameters for the method and validated it. In Ring Study 1, eight laboratories tested the in vitro UV transmission of a total of 24 sunscreens and, from the data, a unit dose of UVA (D(0) of 1.2 J cm(-2)) was defined to provide a single irradiation step which, by taking into account potential sunscreen photo-instability, gave the closest agreement with in vivo UVAPF values. In Ring Study 2, eight laboratories tested the in vitro UV transmission of a total of 13 sunscreens using this single irradiation step and established a very good correlation (r(2) = 0.83; slope = 0.84, P < 0.0001) between resulting in vitro UVAPF values and corresponding values derived from the in vivo PPD method. This new method, therefore, can be used to provide a reliable in vitro metric to describe and label UVA efficacy in sunscreen products, in line with the EU Commission recommendation 2006/247/EC.

Examination of Solar Simulators Used for the Determination of Sunscreen UVA Efficacy

The U.S. FDA recently proposed both in vivo and in vitro UVA efficacy tests for sunscreen products with the lower result used to establish the sunscreen's labeled UVA protection claim. The FDA stated their rationale for dual tests was concern that the in vivo test method overemphasizes UVA-2 (320-340 nm) photoprotection. We attribute FDA's observation to the relative lack, compared to sunlight, of UVA-1 (340-400 nm) radiation in the current JCIA UVA solar simulator specification, allowing the method to generate higher UVA protection factors than sunscreens will provide in sunlight. Our work is based upon comparisons of Air Mass 1.0 sunlight to variously filtered UVA solar simulators. Sources near the JCIA UVA-2/UVA limits (8-20%) had a goodness of fit to solar UVA of only 67-79%. We propose that instead of using ratios of UVA-2 to UVA the standard should be a goodness of fit to the UVA region of an Air Mass 1 solar reference spectrum. As the spectral distribution of solar UVA varies much less than UVB, sunlight of reasonable zenith angles of < or = 60 degrees will have similar spectral shapes and approximate risk spectrum. Goodness of fit to this spectrum will produce UVA protection values predictive to those actually achieved in sunlight of different zenith angles.

Diffuse reflectance spectroscopy for ultraviolet A protection factor measurement: correlation studies between in vitro and in vivo measurements

Assessing the ultraviolet (UVA) protection factor of sunscreen formulations has been discussed for the past 20 years. The purpose of this study is to correlate the measurements of the UVA protection factor value (PFA value) via in vivo diffuse reflectance spectroscopy (DRS) and to compare this method with the in vitro method of measuring the PFA value, as well as with the in vivo persistent pigment darkening (PPD) and PFA methodologies. The UVA protection factor via DRS technique was assessed in two clinical studies. The first study was performed in 12 subjects and the second one consisting of 10 subjects. All subjects in these studies had Fitzpatrick skin phototypes II-IV. DRS measurements were performed using a SkinScan spectrofluorimeter (Spex SkinScan, Yvon Horiba). The in vitro PFA measurements were performed using Labsphere UV-1000s UV Transmission Analyzer. The results obtained from the non-invasive DRS studies were used to correlate with the in vitro testing and with the in vivo PFA/PPD multicenter study. A positive relationship (regression coefficient r(2)=0.90) of PFA values was found between in vitro PFA testing and the in vivo DRS testing. There was also a very good correlation (regression coefficient r(2)=0.99) between the in vivo PFA/PPD values and UVA protection factor obtained from the DRS method. This paper presents studies using the DRS technique to assess the UVA protection factor in different sunscreen formulae correlating with in vitro and in vivo PFA/PPD values. It is a fast method, non-invasive and does not involve any subject irradiation. The technique is a good estimator for the in vivo UVA protection factor as well as a way to assess, in vivo, the photostability of sunscreen formulation in the UVA.