Validation of a new in vitro Sun Protection Factor method to include a wide range of sunscreen product emulsion types.

In 2017, Cosmetics Europe performed a double-blinded ring test of 24 emulsion-type sunscreen products, across 3 in vivo test laboratories and 3 in vitro test laboratories, using a new candidate in vitro SPF test method. Based on the results of this work, an article was published showing how data derived from a new lead candidate method conform to new International Standards (ISO) acceptance criteria for alternative SPF test methods (Any alternative method should consider the matrix effect and if required, specify the matrix applicability of the method; Criterion 1a: Systematic differences between methods should be negligible: 95% of all individual results of an alternative method are within the range of ±2× reproducibility standard deviation of the in vivo method, that is overall bias must be below 0.5× reproducibility standard deviation of the in vivo method; Criterion 1b: Measurement uncertainty of an alternative method should be below the measurement uncertainty of the in vivo method. Candidate method predicted values must fall within the full 'funnel' (SPF 6-50+) limits proposed by Cosmetics Europe (derived from the same minimum test design, that is using the ISO24444 Method to measure at least 24 products across at least 3 laboratories using at least 5 test subjects/laboratory, in a blinded fashion).). Of the 24 sunscreen products tested, the majority of emulsions were of the oil-in-water (O/W) type, whereas only one was water-in-oil (W/O) and there were no products with a mineral-only sun filter system. In order to confirm the scope of this method, therefore, a new study was conducted that included 73 W/O (12 mineral + organic, 44 mineral only and 17 organic only) and 3 O/W mineral-only, emulsion-type sunscreen products (a total of 76 new sunscreen products). When combined with the previous 24 products (tested in 3 different laboratories), this yielded a new data set comprising a total of 100 emulsion-type sunscreen products, with SPF values ranging from 6 to 50+ (with a total of 148 data points). These products were tested using the double-plate in vitro SPF test method and compared with the ISO TC217/WG7 acceptance criteria for alternative SPF test methods. Over 95% of paired in vitro: in vivo SPF values lay within the upper and lower limits of the ISO acceptance criteria funnel, with no bias. This new in vitro SPF test method, therefore, meets the minimum requirements for an alternative SPF test method to ISO24444:2010, for emulsion-type sunscreen products (which make up the majority of marketed sunscreen products).

En 2017, Cosmetics Europe a réalisé un ring test en double aveugle de 24 produits de protection solaire de type émulsion, dans 3 laboratoires de test in vivo et 3 laboratoires de test in vitro, en utilisant une nouvelle méthode de test SPF in vitro. Sur la base des résultats de ces travaux, un article a été publié montrant comment les données dérivées de cette nouvelle méthode sont conformes aux nouveaux critères d'acceptation des normes internationales (ISO) pour les méthodes de test SPF alternatives. Sur les 24 produits de protection solaire testés, la majorité des émulsions étaient du type huile dans l'eau (H / E), tandis qu'un seul était de l'eau dans l'huile (E / H) et il n'y avait aucun produit contenant uniquement des minéraux. Afin de confirmer cette méthode, une nouvelle étude a donc été menée comprenant 73 produits E/ H (12 produits contenant des filtres minéraux + organiques, 44 produits contenant des filtres minéraux uniquement et 17 produits contenant des filtres organiques uniquement) et 3 produits H / E contenant des filtres minéraux uniquement, tous de type émulsion (donc un un total de 76 nouveaux produits de protection solaire). Combiné aux 24 produits précédents (testés dans 3 laboratoires différents), cela a donné un nouvel ensemble de données comprenant un total de 100 produits de protection solaire de type émulsion, avec des valeurs SPF allant de 6 à 50+ (avec un total de 148 points de données) . Ces produits ont été testés à l'aide de la méthode de test SPF in vitro double approche et comparés aux critères d'acceptation de l'ISO TC217 / WG7 pour les méthodes alternatives du SPF in vivo. Plus de 95% des valeurs de SPF appariées in vitro: in vivo se situent dans les limites supérieure et inférieure de l'entonnoir des critères d'acceptation ISO, sans biais. Cette nouvelle méthode de test SPF in vitro, par conséquent, répond aux exigences minimales d'une méthode de test SPF alternative à ISO24444: 2010, pour les produits de protection solaire de type émulsion (qui constituent la majorité des produits de protection solaire commercialisés).

Anti-inflammatory / anti-oxidant activity of ingredients of sunscreen products? Implications for SPF.

OBJECTIVE: The Sun Protection Factor (SPF) of sunscreen products is derived from testing in vivo their ability to prevent erythema ("sunburn"). Recently, certain articles have raised concerns that sunscreen products may actively suppress erythema via anti-inflammatory / anti-oxidant (AI/AO) activity. These articles reason that this may result in a higher labelled SPF value than that provided by the efficacy of the UVR filters alone, giving consumers a "false sense of security". On the other hand, since inflammatory processes are known to play a role in the mechanisms of photodamage / skin cancer induction and propagation, AI/AO activity may provide valuable incremental photoprotective benefit (provided that there is no interference with visible erythema). The objective of these studies, therefore, was to investigate the potential of AI/AO ingredients to suppress UVR-induced erythemal response in human skin, in vivo. METHODS: In vivo studies with SPF30 sunscreen formulations containing a variety of AI/AO ingredients were performed according to the International Standard ISO24444:2010 method. While ISO24444:2010 requires assessment of erythema at 20 ± 4h post-irradiation, an additional assessment at 5 h post-irradiation was also used to determine potential delay in erythema development. RESULTS: None of the formulations, containing a variety of AI/AO ingredients, influenced SPF determination in comparison to the vehicle formulation. CONCLUSION: Our in vivo results demonstrate that commonly-used AI/AO ingredients, at concentrations typically used in sunscreen products, neither influence SPF value nor delay erythemal response, i.e., the measured SPF reflects the true photoprotective capacity of the product.

OBJECTIF: Le facteur de protection solaire (SPF) des produits de protection solaire est dérivé de tests in vivo servant à déterminer leur capacité à prévenir un érythème (« coup de soleil ¼). Récemment, certains articles ont soulevé des inquiétudes en insinuant que les produits de protection solaire pourraient activement faire disparaître un érythème par le biais d'une activité anti-inflammatoire/anti-oxydante (AI/AO). Ces articles soutiennent que cela pourrait impliquer une valeur déclarée du SPF plus élevée que celle fournie par l'efficacité des filtres RUV à eux seuls, donnant ainsi une « fausse impression de sécurité ¼ aux consommateurs. D'autre part, étant donné que les processus inflammatoires sont réputés jouer un rôle dans les mécanismes de photo-altération/d'induction et de propagation du cancer de la peau, l'activité AI/AO pourrait apporter un précieux bénéfice photo-protecteur amplifié (à condition qu'il n'y ait aucune interférence avec un érythème visible). L'objectif de ces études était, par conséquent, d'étudier le potentiel des ingrédients contribuant à l'activité AI/AO à faire disparaître la réponse érythémateuse induite par les RUV dans la peau humaine, in vivo. MÉTHODES: Des études in vivo avec des formules de produits solaires à SPF30 contenant une variété d'ingrédients contribuant à l'activité AI/AO ont été effectuées conformément à la méthode correspondant à la norme internationale ISO24444:2010. Bien que l'ISO24444:2010 nécessite l'évaluation de l'érythème à 20 _ 4 heures post-irradiation, une évaluation supplémentaire à 5 heures post-irradiation a également été utilisée pour déterminer l'éventuel délai d'apparition d'un érythème. RÉSULTATS: Aucune des formules, contenant une variété d'ingrédients contribuant à l'activité AI/AO, n'a influencé la détermination du SPF par comparaison à la formule véhicule. CONCLUSION: Nos résultats in vivo démontrent que les ingrédients contribuant à l'activité AI/AO fréquemment utilisés, aux concentrations généralement utilisées dans les produits de protection solaire, n'influencent pas la valeur du SPF, pas plus qu'ils ne retardent la réponse érythémateuse, autrement dit, le SPF mesuré reflète la véritable capacité photo-protectrice du produit.

Validation of an in vitro sun protection factor (SPF) method in blinded ring-testing.

OBJECTIVE: The objective of this work was to investigate the utility of a new in vitro SPF test method in blinded ring-testing, against new ISO acceptance criteria. METHODS: Twenty four blinded, commercial, emulsion-type, primary sunscreen products, covering the full range of labelled SPF in Europe (SPF6 - 50+), were tested by three test institutes using the current ISO24444:2010 In Vivo SPF Test Method and simultaneously by three separate test laboratories using a new candidate in vitro SPF test method, developed under the leadership of Cosmetics Europe (CE). The resulting relationship between in vitro SPF and in vivo SPF values was then compared with acceptance criteria developed recently by the International Standards (ISO) TC217/WG7 Sun Protection Test Methods Working Group. RESULTS: Analysis of the mean inter-laboratory in vitro and mean inter-institute in vivo SPF values revealed a strong correlation between in vitro and in vivo values, with a Pearson correlation coefficient of r2  = 0.88 (P < 0.0001), a slope of 1.01 and a non-significant intercept (-1.48; P = 0.62). When these data were compared to the new ISO WG7 acceptance criteria, method bias was found to be extremely low and over 95% of the coupled data lay within the model 'funnel' (defined by upper and lower confidence intervals). CONCLUSION: In conclusion, the results of blinded ring testing and comparison to new ISO WG7 acceptance criteria indicate that a new in vitro SPF test method meets (and exceeds) these minimum criteria and is an interesting candidate for possible deployment as an industry test methodology.

The perplexing dilemma of measuring sun protection factors.

Truth, like knowledge, is surprisingly difficult to define. Indeed, every definition of truth that philosophers have developed falls prey to the question, 'Is it true?' In this essay, we consider what is the true SPF of a sunscreen product and whether it can ever be realizable.

A novel proposal for labelling sunscreens based on compliance and performance.

OBJECTIVE: To propose a new way of classifying sunscreens, which combines the labelled SPF with an objective measure of compliance. METHODS: The in vitro determination of uniformity of sunscreen application was combined with an in vivo measure of applied thickness to derive a Compliance Factor for each of 10 sunscreens. RESULTS: The predicted SPF resulting from real-life application of each sunscreen product, which is termed the SPF(in vivo veritas), was calculated from the Compliance Factor and the labelled SPF. It was shown that for a number of products, there is expected to be a significant mismatch between labelled and delivered SPF. CONCLUSION: We believe that by adopting our proposal, consumers would be given more appropriate guidance on the delivered photoprotection they can expect to receive than relying solely on the labelled SPF.

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 (5 µm) and low-roughness (2 µm) plates, were used with a different application rate for each (1.3 mg cm(-2) and 0.75 mg cm(-2) respectively). The UVR dose applied to both plate types followed the same principle as the original test (1.2 J. 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.

A new chemical approach to optimize the in vitro SPF method on the HD6 PMMA plate.

In a previous study, we demonstrated that control of the roughness of molded PMMA plates improves in vitro SPF reproducibility. However, in vitro/vivo deviations are still observed. Sunscreens show different behavior during spreading on the HD6 surface according to the formulation, resulting in a more or less homogenous distribution. The hydrophilic nature of HD6 appears to contribute significantly during spreading. Two different sunscreens offering a homogenous and non-homogenous distribution were investigated to check if the interfacial tension between product and substrate has a real influence on the spreading quality. Using microscopic observations, we attempted to correlate the in vitro SPF results with the product's spreading property. In order to reduce this interfacial tension, an HD6 pretreatment with an amphoteric surfactant, cocamidopropyl betain, was performed. In vitro SPF on "pretreated HD6" was examined using a cohort of 30 products. This pretreatment led to reliable results, demonstrating good association with the in vivo SPF.

Predicting the efficacy of sunscreens in vivo veritas.

A novel in vitro technique for measuring the efficacy of sunscreens over the skin surface is described. It is demonstrated that those products that spread easily are associated with both a low variance in delivered SPF and a subjective assessment of a product that is pleasing to use, which in turn results in a higher application thicknesses and greater delivered photoprotection leading to improved health benefits of sunscreen use.

Validation of HPLC method for the simultaneous and quantitative determination of 12 UV-filters in cosmetics.

The aim of the study was the validation of a high-performance liquid chromatography (HPLC) method for the simultaneous and quantitative determination of twelve commonly used organic UV-filters (phenylbenzimidazole sulfonic acid, benzophenone-3, isoamyl p-methoxycinnamate, diethylamino hydroxybenzoyl hexyl benzoate, octocrylene, ethylhexyl methoxycinnamate, ethylhexyl salicylate, butyl methoxydibenzoylmethane, diethylhexyl butamido triazone, ethylhexyl triazone, methylene bis-benzotriazolyl tetramethylbutylphenol and bis-ethylhexyloxyphenol methoxyphenyl triazine) contained in suncare products. The separation and quantitative determination was performed in <30 min, using a Symmetry Shield(R) C18 (5 microm) column from Waters and a mobile phase (gradient mode) consisting of ethanol and acidified water. UV measurements were carried out at multi-wavelengths, according to the absorption of the analytes.

In vivo persistent pigment darkening method: proposal of a new standard product for UVA protection factor determination.

The European Commission (EC) has recommended assessing the level of ultraviolet A (UVA) protection afforded by sunscreen products using the in vivo persistent pigment darkening (PPD) method or other methods giving equivalent results. In this context, the reproducibility of the in vivo PPD method is of importance. To check the validity of the UVA protection factor (UVAPF) tests, the Japanese Cosmetic Industry Association (JCIA) recommends using a standard product (JCIA standard) with an expected UVAPF 3.75 (SD 1.01). However, considering the increase in UVA efficacy of the new sunscreen products available in the market, with UVAPF up to 30, it seemed useful to develop a new standard product to be used when testing products with expected UVAPF > or =10. The PPD method was used in six centres to determine the UVAPF of the two products. Reproducibility of results was also studied by testing two batches of the new product at two different times. There was no statistical difference between the six centres with regard to the JCIA standard. The ring study showed that the mean value of UVAPF (4.3) was higher than that given by JCIA (3.75). These data enable the proposal of a new acceptance range for the JCIA standard product (3.4-5.2) derived from actual results from European laboratories. Whereas this range is different from that proposed by JCIA (2.74-4.76), there is an overlapping of the values. Data on the new standard product show that reproducibility is not influenced by the batches of this product. The mean UVAPF value obtained is 12.1. An acceptance range (9.6-14.6) is proposed for the new standard. Data presented here demonstrate that if an identical protocol is used, reproducible results can be expected and that the PPD method is reproducible and reliable.

In vitro assessment of water resistance of sun care products: a reproducible and optimized in vitro test method.

The aim of the study was to develop a simple reproducible and reliable in vitro water resistance (WR) method to assess the sun care products. This paper is the result of a scientific collaboration between seven different international industrial laboratories and testing institutes. The same group has already achieved an in vitro protocol for the sun protection factor (SPF) determination [1]. The in vitro WR of sunscreens was tested by applying the same principle as in vivo, which determines the percentage of retention of sunscreen products by assessing the SPF before and after water immersion. Special care was taken to study the parameters influencing the WR and the possibility to follow the kinetics of sunscreen retention during water immersion. The influence of different water qualities has been tested, and osmosed water (1-3 microS cm(-1)) was chosen for the main ring study. Measurement was carried out after 5, 20 and 40 min of immersion. Histograms of selected products demonstrate the percentage of WR at all measuring times and centres, and the regression coefficient to the in vivo determination was shown and statistical calculations clearly demonstrate the reproducibility of the results between the different evaluation centres. The presented method is a practical, convenient and relevant tool for WR screening of sun care and skin care products. It even has the potential to be the starting point for the replacement of the in vivo method in future.

Efficiency of a continuous height distribution model of sunscreen film geometry to predict a realistic sun protection factor.

Irregularities in the geometry of sunscreen films spread on rough areas, like skin, is often presented as being the main cause of the degree of UV absorption achieved by the UV filters that are inside. Until now, only the step film, a model invented by O'Neill, was simple enough to calculate UV data close to in vitro experimental data, after determination of a limited number of fraction areas with their corresponding thickness. However, such models are obviously too simple to represent a real situation. In the present work, more complex distributions of film thickness were calculated, with an infinite number of individual heights. Realistic models were achieved via a probability function. The consequences for UV absorption were deduced, and the calculated UV data were compared to experimental in vitro data on sunscreen products measured after being spread on a roughened PMMA substrate. The latter substrate was previously selected for its ability to achieve a good correlation with in vivo SPF. An optimized version of the continuous mathematical model was finally determined in order to achieve UV curves, similar in shape and intensity to the experimental ones. The latter model can be used to predict realistic SPF values.

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.