UV Filter Substances Research Articles

Experimental and in silico assessment of fate and effects of the UV filter 2-phenylbenzimidazole 5-sulfonic acid and its phototransformation products in aquatic solutions

Often ingredients of personal care products are present in treated wastewaters, e. g grey water (GW), and are discharged into aquatic systems. Conventional treatment of GW does not fully eliminate micropollutants such as the UV filter substance 2-phenylbenzimidazole-5-sulfonic acid (PBSA). Photolysis has been proposed as an alternative treatment method for other micropollutants, but it is not clear yet whether it can also be used to eliminate PBSA. One goal of this study was to better understand the basic pathways involved in this process. It aimed to identify photo-transformation products (PTPs) by using, in the test conditions, an initial concentration of PBSA higher than those expected in the environment. The photolysis experiments were carried out using Xenon and UV lamps. Under Xenon irradiation only slight primary elimination was found. UV irradiation resulted in almost complete primary elimination of PBSA but not in full mineralization. Four isomeric mono-hydroxylated PTPs were identified by high resolution mass spectrometry (HRMS) which could be confirmed by other studies.A modified luminescent bacteria test (LBT) with Vibrio fischeri was employed to assess acute and chronic toxic effects of the irradiated photolytic mixtures. A strong correlation was found between the kinetics of two of the PTPs and luminescence inhibition indicating bacterial toxicity. Using a set of in silico quantitative structure-activity relationship (QSAR) models, this study also offered new insights concerning the environmental fate and toxicity of the TPs of PBSA as the TPs generated by UV-treatment are more persistent and partly more toxic than PBSA.

Photodegradation of the UV filter ethylhexyl methoxycinnamate under ultraviolet light: Identification and in silico assessment of photo-transformation products in the context of grey water reuse

To prevent water shortages in the future and to reduce domestic water consumption, decentralized grey water (GW) reuse has become increasingly important. This water has, however, to be free of pollutants. Conventional treatment of GW does not fully eliminate micropollutants such as the UV filter substance ethylhexyl methoxycinnamate (EHMC). EHMC, which is commonly used in sunscreens and personal care products, is an endocrine disruptor and shows potential to bioaccumulation, which is also reflected in its low water solubility. Photolysis has been proposed as an alternative treatment method for other micropollutants, but it is not clear yet whether it can also be used to eliminate EHMC. One goal of this study was to better understand the basic pathways involved in this process. It aimed to identify photo-transformation products (photo-TPs) by using, in the test conditions, an initial concentration of EHMC higher than those expected in the environment. Acetonitrile (ACN) was added in low concentrations to the aqueous solution to overcome the low aquatic solubility of EHMC. The influence of this co-solvent on the degradation kinetics was studied. The photolysis experiments were carried out using a medium pressure mercury lamp, which emits UV light in the range of 200–400nm. The quantum yield of the photolysis of EHMC was 0.0042 and 0.0023mol·Einstein−1 (for 0.2 and 0.5% ACN (v/v), respectively), and the relative and absolute UV photon fluxes were determined. HPLC was used to monitor the elimination kinetics of EHMC, which followed first-order kinetics. The results of LC-MSn analyses revealed that beside others, several oxidized and hydroxylized EHMC isomers were formed as photo-TPs in aqueous solution. Using a set of in silico quantitative structure-activity relationship (QSAR) models, this study also offered new insights concerning the environmental fate and toxicity of the TPs of EHMC.

Assessment of different biological capacities of Alpinia speciosa (Pers.) B.L. Burtt and R.M. Sm

Objective: The present study was undertaken to evaluate antioxidant, depigmenting and photoprotective activities of the hydroalcoholic extract of Alpinia speciosa leaves. Material and Methods: In vitro spectrophotometric methods were used to determine the total phenolic and flavonoids contents and the above mentioned biological capacities. Results: The antioxidant activity for the hydroalcoholic extract of A. speciosa showed a very good result (IC50=7.13 µg mL-1, statistically different from the ascorbic acid reference standard at p A. speciosa presented IA% lower than 50%, but it also presented a better stability than kojic acid standard. The extract also presented a significant photoprotection for a single UV-filter substance (SPF, UVAPF, Critical Wavelength and UVA/UVB Ratio), and it can be considered a broad-spectrum filter. Conclusion: These data altogether indicate that A. speciosa can have application in cosmeceuticals for aesthetics or degenerative diseases. Key words: Alpinia speciosa, Antioxidant, Depigmentant, Photoprotection.

Fast determination of octinoxate and oxybenzone uv filters in swimming pool waters by gas chromatography/mass spectrometry after solid‐phase microextraction

A fast gas chromatography/mass spectrometry method was developed and validated for the analysis of the potential endocrine disrupters octinoxate and oxybenzone in swimming pool water samples based on the solvent-free solid-phase microextraction technique. The low-pressure gas chromatography/mass spectrometry method used for the fast identification of UV filter substances was compared to a conventional method in terms of sensitivity and speed. The fast method proposed resulted in 2 min runs, leading to an eightfold decrease in the total analysis time and a sevenfold improvement in detection limits. The main parameters affecting the solid-phase microextraction process were also studied in detail and the optimized conditions were as follows: fiber coating, polyacrylate; extraction mode, direct immersion; extraction temperature, 25°C; sample volume, 5 mL; extraction time 45 min; pH 6.5. Under the optimized conditions, a linear response was obtained in the concentration range of 0.5-25 μg/L with correlation coefficients in the range 0.990-0.999. The limits of detection were 0.17-0.29 μg/L, and the recoveries were 80-83%. Combined method uncertainty was assessed and found to be less than 7% for both analytes for concentrations equal to or higher than 5 μg/L. Pool water samples were analyzed to demonstrate the applicability of the proposed method. Neither octinoxate nor oxybenzone were detected in the swimming pool water samples at concentrations above the respective limits of detection.

Photoprotection in changing times - UV filter efficacy and safety, sensitization processes and regulatory aspects.

As excessive sun exposure is tightly associated with different pathological changes of the skin, for example premature skin ageing or the development of skin cancer, an appropriate protection of the skin against UV radiation is of particular importance. Sun protection products and UV filter substances have evolved continuously in the past few decades. New developments and improved technical conditions of production have led to increasingly effective and efficient products with broadband protection ability. Accordingly, legal requirements have also changed and expanded. Although certain trends exist to harmonize the regulation of sunscreens at a global level, there are still large differences how UV absorbers are approved, which testing methods are prescribed, and which general requirements sun protection products must fulfil. Modern UV filters provide efficient protection against UVA and UVB radiation, are heat and photostable, user-friendly, cost-effective, water resistant and non-toxic. As inorganic and organic UV filters are topically applied to the skin in relatively high concentrations (up to 25%), especially the assessment of their (photo)sensitization potential is of particular importance. Accordingly, skin sensitization is a key endpoint for the legally required safety assessment of cosmetic ingredients in Europe and many other countries. This review will summarize the current regulatory status of different approved UV filters, will describe their beneficial and adverse properties and will give an overview of how the efficacy of sunscreens can be evaluated. Finally, an insight into the basic mechanism of (photo)allergic reactions and existing skin sensitization test methods will be provided.

Photostability of cosmetic UV filters on mammalian skin under UV exposure.

Previous studies showed that the common UV filter substances benzophenone-3 (BP-3), butyl methoxydibenzoylmethane (BM-DBM), octocrylene (OCR), ethylhexyl methoxycinnamate (EHMC), ethylhexyl salicylate (EHS) and ethylhexyl triazone (EHT) were able to react with amino side chains of different proteins in vitro. To transfer the results to mammalian skin conditions, sunscreen products were applied on both prepared fresh porcine skin and glass plates, followed by UV irradiation and the determination of depletion of the respective UV filters. Significantly lower recoveries of the UV filters extracted from skin samples than from glass plates indicated the additional reaction of the UV filters with skin constituents, when proteins will be the most important reactants. Among the products tested, BP-3 showed the greatest differences in recoveries between glass and skin samples of about 13% and 24% after 2 and 4 h of irradiation, respectively, followed by EHS > BM-DBM > OCR > EHMC > EHT. The obtained results raise the question, whether the common in vitro evaluations of sunscreens, using inert substrate materials like roughened quartz or polymethyl methacrylate (PMMA) plates are really suitable to fully replace in vivo methods, as they cannot include skin-typical reactions.

Rapid screening method to study the reactivity of UV filter substances towards skin proteins by high‐performance thin‐layer chromatography

Most UV filters used in sunscreens and other cosmetic products contain carbonyl groups, which generally are able to react with peptides or free amino acids of the human skin. To estimate their reactivity, we studied different prominent UV filter substances, octocrylene, ethylhexyl salicylate, 4-t-butyl-4'-methoxydibenzoylmethane, ethylhexyl methoxycinnamate, benzophenone-3, hydroxymethylbenzoyl sulphonic acid, octyldimethyl p-aminobenzoic acid, 3-benzylidene camphor, 4-methylbenzylidene camphor, diethylhexyl butamido triazone and ethylhexyl triazone. A simple screening method using an amino HPTLC plate as protein model was established. The influence of different reaction conditions like heating and irradiation was determined. The ketones BP-3, HMBS and BM-DBM revealed the highest binding rates after both irradiation and heating. After 1h of irradiation, 82%, 28% and 96%, respectively, were bonded to the amino phase, while heating resulted in values of 52%, 36% and 16%. For BP-3 and HMBS, even storage in the dark at room temperature resulted in a low binding. Contrarily, for the two camphor derivatives 3-BC and 4-MBC, only irradiation led to a slightly turnover. UV filters with ester groups also showed a different behaviour depending on their main skeleton. While OCR especially reacted under heating with the amino phase, resulting in 36% of bound species after one hour, UV irradiation particularly encouraged a reaction of the other esters. After 1h irradiation, 15% of EHMC, 38% of EHS and 48% of OD-PABA were bonded to the amino groups of the HPTLC plate, whereas the reactivity of the two triazones, EHT and DEBT, was comparatively low. Especially the UV filters BP-3, BM-DBM, HMBS, EHMC or OCR, which are commonly known to cause contact dermatitis, showed a high tendency to form adducts with the amino layer. Thus, the amino plate seems to be a proper tool to screen for skin sensitizers.

Biotransformation of the UV-Filter Sulisobenzone: Challenges for the Identification of Transformation Products

The UV-filter substance Sulisobenzone (BP-4) is widely employed in sunscreens and other personal care products. In the current study, its behavior during biological wastewater treatment was investigated. In contact with activated sludge BP-4 was degraded in aerobic batch experiments forming at least nine transformation products (TPs). The mass balance in the batch experiments was closed, as measurements with LC-UV underlined that the quantity of the TPs was comparable to the BP-4 quantity transformed. The chemical structures of the nine TPs could be proposed based on accurate mass measurements by high resolution mass spectrometry (LTQ-Orbitrap-MS), several fragmentation experiments up to MS(6) and synthesis of one TP. NMR analyses of the main TP confirmed its proposed chemical structure. At the beginning of the biotransformation of BP-4, a benzhydrol analogue was formed due to the reduction of the keto moiety. Further reactions (e.g., oxidation, demethylation, decarboxylation) led to the formation of extremely polar TPs. A biodegradation pathway was proposed based on the TP structures identified and the sequence of the TP formation. The isolated TPs exhibited higher toxic effects on Vibrio fischeri than BP-4. The results contribute to a better general understanding and prediction of the biotransformation of aromatic sulfonic acids in WWTPs.

Occurrence of widely used organic UV filters in lake and river sediments

Environmental contextPersonal care products containing organic chemicals to filter the sun’s UV rays are produced and used on a broad scale worldwide. Consequently, these organic UV filters are now widespread in the environment. We investigate the occurrence of seven common organic UV filters in river and lake sediments thereby providing valuable data for the future environmental risk assessment of these chemicals to the benthic community of freshwater ecosystems. AbstractPersonal care products (PCPs) are produced and used in huge amounts. These formulations are permanently introduced into the aquatic environment during regular use, mainly through municipal sewage treatment plants. Although there is increasing concern about PCP residues in the aquatic environment, little is known about the extent and level of contamination. The occurrence and concentrations of the seven most frequently used ultraviolet (UV) filters in river and lake sediments have been investigated over a 6-month period by gas chromatography–mass spectrometry: benzophenone-3 (BP-3), 3-benzyliden camphor (3-BC), butyl-methoxydibenzoylmethane (B-MDM), ethylhexyl dimethyl p-aminobenzoic acid (ED-PABA), ethylhexyl-methoxycinnamate (EHMC), 3′-(4′-methylbenzyliden)camphor (4-MBC) and octocrylene (OCR). B-MDM, EHMC and OCR were identified as major contaminants. They were present in every sediment sample with maximum concentrations of 62.2, 6.8 and 642 µg kg–1. 3-BC and ED-PABA could not be detected in any sediment sample. The temporal distribution profile and concentrations of UV filters differed between lakes and rivers. Whereas concentrations of all UV filters in river sediments were low and constant over time, lake sediments exhibited high UV-filter levels during summer and concentrations dropped in autumn. These findings support risk assessment activities and contribute to a better understanding of the magnitude of contamination with organic UV-filter substances in aquatic ecosystems.

Acute and chronic toxicity of four frequently used UV filter substances for Desmodesmus subspicatus and Daphnia magna

As a consequence of growing public concern about UV radiation effects on human health chemical and physical UV filters are increasingly used in personal care and other products. The release of these lipophilic and often persistent compounds into surface waters may pose a risk for aquatic organisms. The aim of the study was to determine effects of four frequently used UV filters on primary aquatic producers and consumers, the green alga Desmodesmus subspicatus and the crustacean Daphnia magna. Exposure to benzophenone 3 (BP3), ethylhexyl methoxycinnamate (EHMC), 3-benzylidene camphor (3-BC) and 3-(4′-methylbenzylidene)-camphor (4-MBC) resulted in growth inhibition of D. subspicatus with 72 h IC10 values of 0.56 mg/L (BP 3), 0.24 mg/L (EHMC), 0.27 mg/L (3-BC) and 0.21 mg/L (4-MBC). EC50 concentrations in the acute test with D. magna were 1.67, 0.57, 3.61 and 0.80 mg/L for BP3, EHMC, 3-BC and 4-MBC, respectively. Chronic exposure of D. magna resulted in NOECs of 0.04 mg/L (EHMC) and 0.1 mg/L (3-BC and 4-MBC). BP 3 showed no effects on neonate production or the length of adults. Rapid dissipation of these substances from the water phase was observed indicating the need for more frequent test medium renewal in chronic tests or the use of flow-through test systems.

The suitability of concentration addition for predicting the effects of multi-component mixtures of up to 17 anti-androgens with varied structural features in an in vitro AR antagonist assay

The risks associated with human exposures to chemicals capable of antagonising the effects of endogenous androgens have attracted considerable recent interest. Exposure is typically to large numbers of chemicals with androgen receptor (AR) antagonist activity, yet there is limited evidence of the combined effects of multi-component mixtures of these chemicals. A few in vitro studies with mixtures of up to six AR antagonists suggest that the concept of concentration addition (CA) provides good approximations of experimentally observed mixture effects, but studies with larger numbers of anti-androgens, and with more varied structural features, are missing. Here we show that the mixture effects of up to 17 AR antagonists, comprising compounds as diverse as UV-filter substances, parabens, perfluorinated compounds, bisphenol-A, benzo(α)pyrene, synthetic musks, antioxidants and polybrominated biphenyls, can be predicted well on the basis of the anti-androgenicity of the single components using the concept of CA. We tested these mixtures in an in vitro AR-dependent luciferase reporter gene assay, based on MDA-kb2 cells. The effects of further mixtures, composed of four and six anti-androgens, could be predicted accurately by CA. However, there was a shortfall from expected additivity with a ten-component mixture at two different mixture ratios, but attempts to attribute these deviations to differential expression of hormone-metabolising CYP isoforms did not produce conclusive results. CA provides good approximations of in vitro mixture effects of anti-androgens with varying structural features.

Pathways of Lateral Spreading

In the case of topically applied substances, usually both lateral spreading and competitive penetration into the skin occur in parallel. In the present study, the pathways of lateral spreading were studied quantitatively and visually. The local distribution and lateral spreading of the UV filter substance butyl methoxydibenzoylmethane applied in an o/w emulsion was studied on the forearm and the back. The tape stripping procedure was used to determine the recovery rates inside and outside the area of application. The skin characteristics of transepidermal water loss, pH value, hydration of the stratum corneum and sebum rate were determined at both anatomic sites. Photography and laser scanning microscopy were used to visually investigate the lateral spreading of topically applied dyes. On the back, a preferred direction of lateral spreading parallel to the body axis was observed. This result was caused by differences in the network of furrows. The furrows functioned as a pathway for lateral spreading, whereas the follicles formed a reservoir for the topically applied substance.

UV-protecting potential of clay minerals and Ganoderma pfeifferi

Exposure with ultraviolet radiation (UV) results in aging of skin and is a leading cause of skin cancer. The cellular responses to UV radiation have received considerable attention over the last few years. Because of the possible phototoxicity of the common physical UV-filter substance titanium dioxide [1] we searched for natural products with UV protecting activities.

A validated method for the determination of traces of UV filters in fish using LC–MS/MS

An analytical method for the determination of UV filter substances in fish tissue has been developed and validated using benzophenone-3, 3-(4-methylbenzylidene)-camphor, 2-ethylhexyl-2-cyano-3,3-diphenyl-2-propenoate and 2-ethylhexyl 3-(methoxyphenyl)-2-propenoate as target analytes. The fish fillets were homogenised and extracted by Soxhlet extraction. The extracts were run through a clean-up process including gel permeation chromatography followed by solid-phase extraction. Quantification of the compounds was performed using liquid chromatography with tandem mass spectrometric detection. Blank fish as well as spiked blank fish were analysed to validate the analytical method. The analytical method developed has the multiple advantages of enabling separation, simultaneous identification and quantification of each of the four selected compounds in a single run. Contamination of blank samples and abnormally high concentrations in spiked samples were avoided by taking extensive precautions during the fish preparation procedure. The method was validated in accordance with internationally accepted criteria, such as specificity, accuracy and repeatability. The combination of LC with tandem mass spectrometry ensures a high level of specificity. The accuracy of the method was reported as the mean recovery rate for the analytes in the sample matrix. Mean recoveries were in the range 86-108%. The precision is expressed as the relative standard deviation, and in all but one of the cases was 20% or below. The accuracy of the method allows residue analyses to be performed on biological matrices at ng/g levels. The determined limit of quantification for each analyte was 8 ng/g fish. For all spiking levels > or =8 ng/g, relative standard deviations were < or = 20%.

Estimation of the relative stratum corneum amount removed by tape stripping

The tape stripping procedure is a suitable minimal invasive tool to study, e.g. the penetration and dermatopharmacokinetics of topically applied substances. In the present study, this procedure was used to remove the stratum corneum (SC) completely and to study the penetration of the UVA filter substance butyl methoxydibenzoylmethane after application in two different vehicles. The amount of corneocytes removed by each tape strip from the flexor forearm of human volunteers was determined via their pseudo-absorption. In a second part, the penetration profiles of a UVA filter substance applied in two different vehicles were determined following the developed standard protocol using the tape stripping procedure in combination with UV/VIS spectroscopy. The amount of corneocytes removed by each tape strip was related to the number of tape strips used for removal. Mean values with a deviation of less than 20% concerning the relative amount of SC removed by a constant number of tape strips were obtained. For instance, a relative amount of 66 +/- 12% was removed with the first 20 tape strips, while nearly the complete SC (95 +/- 3%) was removed using 50 tape strips. In addition, these results were used to estimate the relative SC amounts removed, studying the penetration of the UVA filter substance after application in two different vehicles. No significant differences between the distributions of the UV filter substance applied in both emulsions were obtained (P > 0.05). The reported procedure for the estimation of the removed SC amount provides the possibility to avoid the complete removal of the SC and to compare the penetration characteristics obtained for different volunteers and different products in relation to the relative horny layer profile.

Determination of penetration profiles of topically applied substances by means of tape stripping and optical spectroscopy: UV filter substance in sunscreens

Penetration profiles of topically applied drugs and cosmetic products provide important information on their efficacy. The application of tape stripping in combination with UV/VIS spectroscopy is checked to determine the local position of topically applied substances inside the stratum corneum, the penetration profile. The amount of corneocytes removed with each tape strip is quantified via the particle-dependent absorption, the pseudoabsorption, in the visible spectral range. The concentration of a typical UV filter substance, 4-methylbenzylidene camphor, is determined by optical spectroscopy using the tape strips removed originally. In this case, a time-dependent increase in the absorbance must be taken into account. Laser scanning microscopic investigations confirm that the nonhomogeneous distribution of the filter substance, on the strips, can explain this spectroscopic behavior. When reaching a homogeneous distribution, the UV spectroscopic signal reflects the correct concentration. These spectroscopic values are compared with high performance liquid chromatography (HPLC) data. The values obtained with both methods for the concentrations of 4-methylbenzylidene camphor are in good agreement. The data obtained are used to illustrate the determination of a penetration profile of a UV filter substance. The results demonstrate that the described protocol is well suited to characterize, in a simple manner, topically applied substances that have a characteristic UV/VIS absorption band.

Synergy effects between organic and inorganic UV filters in sunscreens

The influence of the synergy effects between organic and inorganic UV filter substances on the sun protection factor (SPF) of topically applied sunscreen formulations is investigated. The medium is considered to have reflection, absorption, and scattering properties. The distribution of photons in this medium is investigated by Monte Carlo calculation. Typical optical parameters of the skin and substances are used to characterize the synergy effect. The results of the model calculation are checked by in vitro and in vivo measurements investigating the influence of different types of scattering microparticles on the absorption efficacy of topically applied formulations. It is found that the inorganic filter substances act as scattering microparticles in the upper skin layers. They increase the optical pathway of the photons in the topically applied absorbing formulation also localized there. In this way, more photons are absorbed, increasing the SPF. The results obtained are important for the optimization of the SPF of sunscreen formulation containing organic and inorganic UV-filter components.

Orally administered ethanol: transepidermal pathways and effects on the human skin barrier

Ethanol intake is associated with a variety of skin diseases. The aim of the present study was (1) to identify the pathways of release of orally administered ethanol through the skin, and (2) to investigate the effects of a single oral dose of ethanol on the penetration of topically applied substances into the skin. Ethanol evaporation via the skin was measured using the new technique of ion mobility spectrometry (IMS). Transepidermal water loss (TEWL) and skin surface temperature were simultaneously measured before and after ethanol consumption. Measurements were performed on skin sites with different stratum corneum (SC) thickness, and density of follicles and sweat glands. These appendages were selectively sealed to investigate their participation in ethanol evaporation. The penetration of a topically applied UV filter substance was studied before and after ethanol consumption after removing the SC with adhesive tape. Ethanol evaporation was measured within 5 min of consumption, while the skin surface temperature remained nearly constant. The sealing of the appendages did not have a significant effect on ethanol evaporation. On the forehead, a higher TEWL value was measured than on the forearm. On both skin sites, an increase in TEWL was observed after ethanol ingestion. No influence of orally administered ethanol on the penetration of the topically applied UV filter substance was observed. The results indicate that ethanol evaporation occurs via the lipid layers without a significant effect on the penetration of the topically applied substance.

Lateral Spreading of Topically Applied UV Filter Substances Investigated by Tape Stripping

The lateral spreading of topically applied substances is a competitive process to the penetration into the stratum corneum (SC). The penetration of topically applied UV filter substances into the human SC and the lateral spreading were investigated in vivo. Tape stripping in combination with spectroscopic measurements was used to study both processes of two UV filter substances. The concentration of both UV filters was determined inside and outside the application area by varying the application and tape stripping protocol. A spreading of the topically applied substances from the treated to the untreated areas was observed, which caused a concentration gradient. This lateral spreading depends on the time between application and tape stripping and the size of the treated skin area. Significant amounts of topically applied substances were found adjoining the application area, due to the lateral spreading which takes place on the skin surface. In general, the lateral spreading must be considered to be a competitive process when studying penetration processes of topically applied substances. It has to be considered during drug treatment of small limited skin areas and for the interpretation of recovery rates obtained in penetration studies.

Distribution of sunscreens on skin

The effectiveness of sunscreens was originally achieved by incorporation of soluble organic UV absorbers such as cinnamates and others into cosmetic formulations. Determinations of the sun protection factor (SPF) of emulsions containing different organic UV absorbers clearly indicate that the efficacy depends on the absorption characteristics of each single UV filter substance. Nowadays, micronised pigments such as titanium dioxide or zinc oxide have also been found to be protective against harmful UV rays. Our investigations using optical and electron microscopy proved that neither surface characteristics, particle size nor shape of the micronised pigments result in any dermal absorption of this substance. Micronised titanium dioxide is solely deposited on the outermost surface of the stratum corneum and cannot be detected in deeper stratum corneum layers, the human epidermis and dermis.