Beyond sun protection factor testing

Ultraviolet radiation in sunlight produces a range of acute and chronic adverse effects on the skin including sunburn, photosensitivity rashes, immunosuppression, photoageing and carcinogenesis. Sunscreens aim to provide protection, but standard testing procedures primarily involve assessment of ability to protect against acute erythema, as evidenced by the sun protection factor (SPF). The SPF may correlate poorly with other aspects of protection, particularly since ultraviolet A is weakly erythemogenic compared with ultraviolet B, yet may make a greater contribution to certain other skin effects of sunlight. Nevertheless, there is an increasing tendency for the sunscreen industry to make claims for their products beyond the SPF data. There is a need to develop systems for clinical testing of sunscreens against other endpoints caused by ultraviolet exposure of skin, including immunosuppression and photosensitivity rashes. In particular, there is a largely unrecognized need for testing of sunscreens against the condition known as polymorphic light eruption, a photosensitivity disorder estimated to affect a staggering 10-20% of the population in the northern hemisphere. Ultimately, protection of the skin by sunscreens can only be as effective as their adequacy of application to the skin surface in the everyday setting permits. Optimal sunscreen formulation, and public and patient education in appropriate application technique, both make vital contributions to efficacy of sunscreen protection. This article focuses on the need for extended clinical testing of sunscreens, with particular reference to the photosensitivity disorders, and for improvements in sunscreen formulation and in the adequacy of sunscreen application to the skin surface.     

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.     

New data on skin photoprotection

Absorbed ultraviolet (UV) radiation is responsible for photochemical degradation of the skin. Erythema is the most apparent result of the sunburn reaction, but the most serious results are photoaging and photocarcinogenesis, largely mediated by reactive oxygen species (ROS). To prevent these light-induced reactions, effective protection against both UVB and UVA radiations are essential. The aim of this study was to evaluate the protective efficacy against both UVB and UVA rays of three sunscreen preparations (two creams and one lotion), evaluating also their water resistance. For this purpose SPF (Sun Protection Factor) and UVA protection were tested, respectively, on 40 and 20 pre-selected volunteers (male and female, < 22-years-old with skin types I, II and III) by the Colipa test method and the Mark et al. methodology. A wash-off test was also conducted to determine the resistance to water. Results obtained showed that the three formulations studied underline interesting SPF values from about 20-30 with an excellent correlation between visual and colorimetric results obtained (P < 0.05). The controlled decrease in effectiveness of only 20% after 10 min washing proved that the vehicle is also an important factor in determining the protective efficacy. From the obtained data, the studied formulations seem to have an high SPF and UVA protection efficacy together with a good water resistance. According to several other studies, it is possible to confirm therefore that the regular use of these sunscreens may certainly reduce the harmful effect caused by long-term exposure to UVB and UVA radiation.     

Sunscreen in vitro spectroscopy: application to UVA protection assessment and correlation with in vivo persistent pigment darkening

In the present study, we have described an in vitro spectroscopic method to evaluate the sunscreen products for UVA sun protection factor. The roughened PMMA plates have been used as a transparent substrate on to which the test product is spread. The UVA protection factors have been deduced from the UV-transmittance data measured in the UVA area. In order to be as close as possible to the in vivo protection factors, issued from the PPD end-point, the treated polymethylmethacrylate (PMMA) plates are submitted to different UV-irradiation doses, before the measurement. The correlation in vitro/in vivo is poor when the sunscreens are not irradiated. A UV dose of about 2 minimal pigmenting dose (MPD) is enough to achieve a good correlation between in vitro and in vivo data issued from the 13 tested sunscreens. These results are consistent with the fact that the photostability of sunscreens is challenged during an in vivo PPD test.     

A novel proposal for the assessment of sunscreen product efficacy against UVA

The ideal characteristics are outlined for an assessment method for the protective properties of sunscreen products against UVA. Existing sun protection factors (SPFs) are reliant on largely UVB-induced delayed erythema, which is inappropriate for the UVA. A novel instrumental method for assessing a protection index is detailed which is applicable to the UVA. The validation of the method relies on measurements in the UVB and comparison with conventional SPFs, since no reliable UVA protection index exists. The method uses UV transparent skin casts to which products can be applied. Throughput of appropriately filtered light, with and without applied product, is measured to generate a protection index. Comparison with SPFs confirms that the measurements are similar to those generated by in vivo methods, but that careful choice of the casting medium is essential.     

Spectral uniformity: a new index of broad spectrum (UVA) protection

Despite the wide acceptance of in vitro assay for the determination of the spectral absorbance of a sunscreen product, we find that different authorities are recommending different summary metrics of this spectral profile to express the UVA or broad spectrum, protection offered by topical sunscreens. Regrettably, the situation that now prevails is not ideal and the option for rationalizing the situation in the interests of consumer-focussed international harmonization is either the adoption of one of the existing metrics or universal acceptance of a new metric for expressing broad spectrum (UVA) protection. There may be greater harmony in adopting the second proposal and so I introduce here a new metric, which I term the spectral uniformity index , for consideration by the international sunscreen community. It is shown that this new index is aligned to the fundamental requirement of a topical sunscreen to provide a flat spectral profile, is simple to calculate and conceptually easy to understand, and correlates strongly with existing indices.     

In vitro testing to assess the UVA protection performance of sun care products

The UVA protection delivered by sunscreens is an issue of increasing importance due to the increasing knowledge about UVA-induced skin damage. In Europe there is no officially accepted method available to determine the degree of UVA protection. Therefore, the objective of the present study was to design a protocol combining the merits of an in vitro model, which are simple and reproducible, with aspects known to be relevant from in vivo studies. The principle is: an UV-transparent support to which the test product is applied, a (pre)irradiation and a transmission measurement. Transpore(R) tape (standard support for SPF determinations) was found to be incompatible with many preparations on prolonged contact times. Roughened quartz was adopted as a suitable alternative. Transmission measurements on this support are not reliable with a layer of 2 mg cm(-2) (standard for SPF) due to detection limitations of spectrophotometers, hence a reduced layer of 0.75 mg cm(-2) was adopted. Overall, it is very difficult to apply products in a reproducible thin layer on appropriate substrates. As a consequence, absolute parameters derived from the transmission profile show relatively large dispersion, whereas relative parameters, such as critical wavelength lambda(c)[1] or UVA/UVB ratio are much less sensitive to unavoidable variations in layer thickness. An increase in deviations was observed when the samples were irradiated before measurement. It is crucial to control the output carefully (spectral distribution and even more importantly, irradiance and dose delivered) of the light source. By doing so and also taking into account the previous learning steps, a protocol was drafted and tested in a ringtest (four samples in six laboratories). The results are encouraging and show that if relative parameters (e.g. lambda(c), UVA/UVB ratio) are considered, the intra- as well as interlaboratory reproducibility is clearly better than can be obtained in vivo. In general, we describe a suitable method, which can be considered in any future official discussions about the methodology to determine UVA protection.     

Consequences for sun protection factors when solar simulator spectra deviate from the spectrum of the sun

The sun protection factor (SPF) of two products, one with an expected SPF of 4 and another with an expected SPF of 15 were determined, using two solar simulators: Multiport Solar UV Simulator (xenon, Solar Light, Philadelphia, PA, USA), and Supersun 5000 (metal halide, Mutzhas, Munich, Germany). The mean SPFs using the Multiport were: 4.8 for the low SPF product and 19.4 for the high SPF one. The results using the Supersun were lower: 2.6 for the low SPF product and 7.2 for the high SPF one. Relative emission spectra of the two sources were recorded using a fluorescence spectrophotometer in bioluminescence mode. Efficacy spectra were calculated and compared with the corresponding spectrum of natural sunlight. It was evident that the spectral power of the xenon source is too high in the UVB, leading to overestimation of SPFs, whereas the Supersun irradiator emits too much in the UVA, resulting in too low SPFs. Heat effects and photodegradation of UV filters are discussed as further possible reasons for the discrepancies between the experimentally determined SPFs. Our results confirm a recent publication about theoretical SPFs, calculated with emission spectra of a xenon source and spectra of the sun at different elevation angles, where the authors provide evidence that in natural sunlight the contribution of UVA to total UV radiation is twice as high as in a xenon source. This may contribute to an understanding of why sunscreens tested according to the FDA method (xenon sources) often yield higher SPFs than those obtained from European testing procedures.     

Comparative evaluation of different substrates for the in vitro determination of sunscreen photostability: spectrophotometric and HPLC analyses

Polymethylmethacrylate (PMMA) plates and Transpore^TM tapes were compared as substrates for the in vitro evaluation of photostability of commercial sunscreen products. The sun care preparations were applied respectively on Transpore^TM tapes and PMMA plates and their sun protection factors (SPF) and UVA protection parameters [UVA/UVB ratio, critical wavelength, UVA protection factor (UVA‐PF)] were measured by transmission spectroscopy, before and after irradiation with simulated sunlight. No significant differences were observed in the UV protection parameters measured on Transpore^TM tapes or PMMA plates, before exposure to the solar simulator. Conversely, after irradiation, the SPF values of the sun care products exhibited marked variations between the two substrates, the decrease in SPF being greater on PMMA plates (31.3–63.1%) than on Transpore^TM tapes (10.4–23.8%). Differences between the two substrates were detected also for the UVA protection parameters, although they were significant only for the UVA‐PF. The tested samples were assayed also by high‐performance liquid chromatography (HPLC) to assess the extent of photodegradation of the UV filters present in the examined formulations. The results showed that for the PMMA plates, the light‐induced decrease in SPF, as determined by spectrophotometry, fitted well with the percentage loss of ethyl hexyl methoxycinnamate (the only photounstable UVB filter present) measured by HPLC. Moreover, for the PMMA substrate, the UVA‐PF percentage reduction was consistent with the percentage degradation of butyl methoxydibenzoylmethane (the only photounstable UVA filter present) determined by HPLC. On the other hand, poor correlation between spectrophotometric and HPLC analyses was observed on Transpore^TM tapes. Therefore, PMMA plates are more reliable than Transpore^TM tapes as substrates for in vitro photodegradation tests of sunscreen products by transmission spectroscopy.     

Comparison of sun protection factors determined by an in vivo and different in vitro methodologies: a study with 58 different commercially available sunscreen products

An extensive study on the sun protection factors (SPF) of sun care products was carried out using the COLIPA (The European Cosmetic Toiletry and Perfumery Association) method, which relates to in vivo experiments. Furthermore, in vitro methods were tested with sunscreen formulations that were prepared as films on surface-roughened plates of polymethyl methacrylate (PMMA). One of the in vitro methods, i.e. using the sunscreen tester, has been recently developed, whereas the second has been defined by a pure spectroscopic approach, which is based on spectral transmission measurements of sunscreen films. Altogether 58 different sunscreen formulations, with manufacturer declared SPF values ranging from 4 to 60 and currently available on the European market, were investigated. The quality of correlations with results from the individual products based on the different in vitro methods versus the COLIPA values that were considered as generally accepted standard values was assessed. In this context, also variations because of sample preparation and spectral measurement were discussed. For sunscreen products with in vivo SPF values larger 25, the spectral transmittance within the UVA/UVB range is rapidly decreasing, which is experienced even for products with reduced amounts reaching 0.5 mg cm(-2) and still leading to unsatisfactory correlation of the spectroscopically derived SPF values versus the results from the alternative assays. Opposite to these small amounts, a sunscreen product spread of 2 mg cm(-2) is standard for the in vivo COLIPA method, whereas an area-normalized amount of 1 mg cm(-2) is currently routinely used for the sunscreen tester method. Furthermore, an overview of the individual product characteristics, such as their specific critical wavelengths and their UVA/UVB ratios is provided; both parameters can also be calculated from the spectral absorbances of the standardized sunscreen films.     

In vivo and in vitro assessment of UVA protection by sunscreen formulations containing either butyl methoxy dibenzoyl methane, methylene bis-benzotriazolyl tetramethylbutylphenol, or microfine ZnO

The UVA-attenuating properties of the three UVA filters butyl methoxy dibenzoyl methane (BMDBM), methylene bis-benzotriazolyl tetramethylbutylphenol (MBBT), and microfine zinc oxide (ZnO), are compared. For this purpose persistent pigment darkening (PPD) as an in vivo method as well as different in vitro approaches like the UVA/UVB ratio, the critical wavelength, and the Australian standard have been used. For the case of the UVA/UVB ratio and the critical wavelength the behaviour was also assessed after irradiation with 10 minimal erythemal doses (MED). Sunscreen formulations were manufactured containing either one of these UVA filters or combinations of one UVA filter and a constant amount of UVB filter. The concentration of the respective UVA filter was varied. BMDBM was dissolved in the oil phase of the formulations. MBBT, which is produced as particulate organic UVA-filter dispersion with average particle size smaller than 200 nm, was added to the aqueous phase. The microfine ZnO was incorporated into the oil phase. If no additional UVB filters were present, UVA/UVB ratio and critical wavelength stayed almost constant, independent of UVA-filter content. With constant levels of additional UVB filters these parameters increased with UVA-filter concentration. In contrast to the behaviour of the UVA/UVB ratio, which appeared to be a sensitive measure for the UVA protection in relation to UVB protection in almost the whole ranges of UVA-filter concentrations, the critical wavelength approached saturation already at low UVA-filter levels. The UVA-protection factors (UVA-PF) obtained from the in vivo studies increased with the concentration of the UVA filter in the formulations. Formulations, which showed UVA-PFs > or = 4 in most cases met also the conditions of the Australian Standard. An irradiation dose of 2.5 kJ m(-2) (10 MED) induced significant decreases of UVA/UVB ratio or critical wavelength only with some BMDBM formulations, indicating a loss of UVA protection in those cases.     

Reconstructed human epidermis as an efficient tool in the evaluation of the effects of UV irradiation and of the photoprotective capacities of a sunblock

In this study, we used a spectroscopic method, found effective in measuring sunscreen photo-degradation results in terms of a photoabsorption decrease. The traditional approach was to measure variations in well-known parameters such as the absolute in vitro sun protection factor (SPF) or the relative UVA/UVB ratio after a controlled dose of UV radiation. However, these parameters fail to fully reflect the product's photochemical behavior. Variation in the SPF mainly depicts changes in UVB filtration, and variation in the relative parameter UVA/UVB ratio can be subjected to misinterpretation, as is the case for products in which a parallel loss of photoabsorption is observed in the UVA and UVB regions. In our laboratory, we developed a new approach using pure spectroscopic UVA and UVB attenuation factors called UVA attenuation factor and UVB attenuation factor. Attenuation factors are defined here as the inverse of the arithmetic mean of the transmittance data, taken either from the UVB region (290–320 nm, 1 nm steps) or the UVA region (320–400 nm, 1 nm steps). Using these two new spectroscopic indices, the photochemical behavior of a sunscreen formula can be assessed in each UV region. The general procedure consists of applying two UV doses to a film of sunscreen previously spread on a roughened polymethylmethacrylate plate and then measuring the UV transmission versus wavelength before and after the irradiation process. We tested different UV filters in a cosmetic base to demonstrate the efficacy of the method.     

Research on a natural sunscreen from Chinese herbs

To compare the difference in SPF values between a synthetic sunscreen and a sunscreen derived from Chinese herbs, by using optical and biological methods, we observed the effects of these sunscreens by photochemical reaction, sun protection factor (SPF) values, and UV absorption. The results showed that the synthetic sunscreen was more effective in UV absorption than the sunscreen derived from natural Chinese herbs. When they were tested biologically, it was found that the sunscreen derived from Chinese herbs was able to improve skin tonicity, as well as reduce the itching, photoerythema, squamation, burning, and reddening caused by the excessive UV radiation, while the synthetic sunscreen could not improve these conditions of the skin, and even induced some side-effects in the skin with photoerythema. By comparing the in vivo SPF values of the two types of sunscreen, they were found to be mainly similar. The results indicate that the SPF value was related to both a photo-reaction and a biological reaction. The sunscreen derived from natural Chinese herbs, although having low UV absorption, was able to exhibit the same SPF value as the synthetic sunscreen because of its biological effects.     

Poly(D,L-lactide) nanoencapsulation to reduce photoinactivation of a sunscreen agent

The use of sunscreens is the 'gold standard' for protecting the skin from ultraviolet light. Octyl methoxycinnamate (OMC) is one of the most widely used UVB filter but it can act as a sensitizer or photoallergen. When exposed to sunlight, OMC can change from the primary trans-form to cis-form and the isomerization, not reversible, conducts to a reduction of the UVB filtering efficiency because the trans-form has a higher extinction coefficient. Photostability is the most important characteristic of effective sunscreens and it can be influenced by formulation ingredients and by applying technological strategies. In this work, photostability experiments, performed on emulsion-gels containing different percentages of OMC free or loaded in poly(D,L-lactide) nanoparticles, were carried out. The presence of a polymeric envelop may act to protect the active ingredient. In this study, the influence of poly(D,L-lactide) matrices on the photochemical stability of the sunscreen agent was investigated. As highlighted in this study, free OMC in different formulations has different photoisomerization degree. Moreover, a dissimilar behaviour was observed by studying different sunscreen concentrations in the same cosmetic formulation. Photostability results show a significant reduction in photoisomerization degree for formulations containing sunscreen loaded in nanoparticles, highlighting that the encapsulation is a suitable strategy to improve OMC photostability. Moreover, sun protection factor (SPF) results show that the UVB filter protective power is also maintained after encapsulation.     

In vitro testing of zinc oxide sunscreens

The UVA performances of two all-mineral zinc oxide sunscreens are measured following Colipa and ISO procedures and compared to a sunscreen containing only organic actives. It is found that the two sunscreen types yield very different in vitro SPF and UVA results. It shown that the results can be rationalized in terms of the differences in the monochromatic extinction spectra of the two types of sunscreens.     

In vitro evaluation of sun protection factors of sunscreen agents using a novel UV spectrophotometric technique

A method for the in vitro determination of low- and high-value sun protection factors (SPF) of sunscreens using artificial substrates and a novel pseudo double beam (PDB) mode of operation of a standard double beam UV spectrophotometer is described. The method allows transmittance to be calculated from detector responses of reference and sample beams measured at different gain levels and facilitates the accurate quantification of low levels of electromagnetic radiation transmitted through highly absorbing samples. The spectrophotometer was modified to hold quartz diffusing plates on which a substrate [Transpore™ adhesive tape or human stratum corneum obtained from a skin surface biopsy (SSB)] and the sunscreens to be tested were applied. The PDB mode of operation increased the effective linear range of the detector response of the spectrophotometer by a factor of approximately 20000-fold, enabling the in vitro SPF determination technique to be applied to both high and low SPF value sunscreens. Eight commercial sunscreens with known SPF values ranging from 4 to 77, previously determined by in vivo methods, were tested in vitro using both test substrates and correlations between the in vivo and in vitro values were determined. SPF values determined using the in vitro method correlated well with the known in vivo results (Transpore™ tape, R ² = 0.611; SSB, R ² = 0.7928). The in vitro SPF obtained for one of the tested products differed substantially from the cited in vivo SPF value. Independent in vitro and in vivo re-evaluation of the SPF of this product matched the value predicted by the present method much more closely than the originally cited in vivo value. All determined SPFs were ordered correctly in comparison to in vivo ranking and the technique appeared to correctly identify a sunscreen that had a labelled SPF value that was significantly higher than its true SPF.     

Beyond UV radiation: A skin under challenge

Since ancient times, human beings have been trying to protect their skin against the adverse effects of the sun. From the first mineral sunscreens used by Egyptians, to the current more sophisticated ultraviolet (UVA/UVB) organic sunscreens, progress has been made in terms of sun protection and deeper knowledge of skin physiology has been acquired in the process. The solar spectrum is composed of radiations of various wavelengths having specific, as well as overlapping effects on skin. UVB is mainly responsible for sunburn and DNA dimer formation that can lead to mutation. UVA generates oxidative reactions affecting DNA, proteins and lipids, and is also immunosuppressive. Recently, visible light and infrared radiation (IR) have been associated with oxidative damage and IR has been additionally linked to adverse heat effects on skin. Numerous other extrinsic factors, related to environment and lifestyle, also affect the appearance of skin, precipitating ageing. New molecular mechanisms linking sun and environmental factors to skin ageing have been identified: IR affects mitochondrial integrity and specific heat receptors also mediate some of its effects, tryptophan is a chromophore for UVB, and the aryl hydrocarbon receptor (AhR) is activated by light and xenobiotics to alter skin physiology. Integrating all these new elements is changing the way we think about skin extrinsic ageing. Is UVA/UVB sunscreen protection still enough for our skin?     

A National Web-Based Survey of Sunscreen Products as a Tool for Industry Self-Regulation, Consumer Awareness Campaigns and Marketing

The objective of the work was to test a web-based tool in the collection and presentation of data for our regular sunscreen surveys and to develop, test and conduct a web-based survey of sunscreen products, with instant data reporting. After the Photobiology Laboratory at MEDUNSA was approached by SurveyIT to pilot a new web-based data collection and reporting tool, companies were contacted via e-mail and invited to visit a secure web-based site to complete the provided questionnaire. Data collection and processing were greatly simplified by the new approach. The survey gleaned responses from 18 companies on 31 brands with a total of 110 primary sunscreen products. The data revealed a reduction in the number of companies marketing sunscreens in South Africa compared with the previous year, despite an increase in the volume of sunscreen sales. Ninety-eight percent of the products were claimed to be in vivo SPF tested. Fifty-seven percent of the products had undergone in vitro SPF testing for determination of their UVA protection. Fifty-one percent of the products were claimed to be water-resistant. Octyl methoxycinnamate was the most popular UV filter (88% of products), followed by benzophenone-3 (60%). Butyl methoxydibenzoylmethane was in 52% of the products and surprisingly only 29% of the products contained titanium dioxide. The major conclusions are that the data collection tool was a success and that sunscreens are becoming more widely used and more thoroughly tested in South Africa.     

Adaptation of the protocol for determining in vitro the sun protection factor of anti-solar sticks

Apart from the protection offered by clothing, the application of sunscreen products suited to each type of skin constitutes one way for decreasing the frequency of skin cancers nowadays. After having adapted an in vitro method for determining the efficacy of sunscreens in emulsion form, we wished to transpose this technique by adapting it for the anti-solar sticks for the evaluation of sun protection factor (SPF) using a spectrophotometer equipped with an integrating sphere. To do this, we tested 14 products in the market as well as sticks that we ourselves fabricated in the laboratory. In a base common to all of these sticks, we added organic (13 filters tested) and inorganic (two filters tested, titanium dioxide and zinc oxide) to their maximum permitted concentration in the European Union. In parallel, emulsions containing the same filters at the same percentage of use were studied; to be in keeping with the results on the products packaging on the one hand, and with the results obtained for the emulsion form on the other hand, we were able to determine the optimal mass which needed to be placed on the support used the in vitro test to determine the SPF.     

New ex-vivo method for evaluating the photoprotective efficacy of sunscreens

The photoprotective efficacy of some topical sunscreens was determined ex-vivo by spectrophotometric assay using an irradiation monochromator and a diffuse reflectance sphere. Transmission scans were run from 400 to 290 nm through human epidermis samples before and after application of products and protection factors (PF) were calculated. The results are also expressed as the percentage of the radiation stopped by the products in each spectral range (UVB, short UVA and long UVA). All the formulations contained two chemical filters: butyl methoxy dibenzoyl methane and octyl methoxycinnamate in a range of concentrations. Estimates of the effectiveness of all products were higher than results in humans when calculated by this ex-vivo technique. Due to its high discriminant power, this method constitutes a useful tool in the formulation of sunscreens.