Study on the photostability of guaiazulene by high-performance liquid chromatography/mass spectrometry and gas chromatography/mass spectrometry

The photostability of guaiazulene (1,4-dimethyl-7-isopropylazulene; GA), a natural azulenic compound used in cosmetic and health-care products, as well as in pharmaceutical preparations, was investigated in solution (methanol, ethanol, acetonitrile), by different techniques: gas chromatography/mass spectrometry (GC/MS) and high-performance liquid chromatography combined with atmospheric pressure chemical ionization mass spectrometry and UV detection (LC/APCI-MS and HPLC/UV). A solar simulator (xenon-arc lamp) was used as UV-A radiation source. The study involved: monitoring compound decomposition, identifying products of photodegradation (PPs), assessing the role of oxygen and evaluating the kinetics of the process. Minor PPs are volatile compounds and were characterized by GC/MS, while oligomeric polyoxygenated compounds, tentatively characterized on the basis of MS and MS/MS spectra, were found to be the main photoproducts. The photodegradation was found to be enhanced by the presence of oxygen; nevertheless, determination of the singlet oxygen quantum yield for GA gave a lower value than that for the reference standard Rose Bengal. The obtained results and the developed stability-indicating methods (GC/MS and LC/MS) are of interest for stability studies and/or quality control purposes of GA as raw material or cosmetic products.     

An Overview of the Analytical Methods for the Determination of Organic Ultraviolet Filters in Cosmetic Products and Human Samples

UV filters are a group of compounds commonly used in different cosmetic products to absorb UV radiation. They are classified into a variety of chemical groups, such as benzophenones, salicylates, benzotriazoles, cinnamates, p-aminobenzoates, triazines, camphor derivatives, etc. Different tests have shown that some of these chemicals are absorbed through the skin and metabolised or bioaccumulated. These processes can cause negative health effects, including mutagenic and cancerogenic ones. Due to the absence of official monitoring protocols, there is an increased number of analytical methods that enable the determination of those compounds in cosmetic samples to ensure user safety, as well as in biological fluids and tissues samples, to obtain more information regarding their behaviour in the human body. This review aimed to show and discuss the published studies concerning analytical methods for the determination of organic UV filters in cosmetic and biological samples. It focused on sample preparation, analytical techniques, and analytical performance (limit of detection, accuracy, and repeatability).     

In Vitro Evaluation of the Photoreactivity and Phototoxicity of Natural Polyphenol Antioxidants

Polyphenols are a large family of natural compounds widely used in cosmetic products due to their antioxidant and anti-inflammatory beneficial properties and their ability to prevent UV radiation-induced oxidative stress. Since these compounds present chromophores and are applied directly to the skin, they can react with sunlight and exert phototoxic effects. The available scientific information on the phototoxic potential of these natural compounds is scarce, and thus the aim of this study was to evaluate the photoreactivity and phototoxicity of five phenolic antioxidants with documented use in cosmetic products. A standard ROS assay was validated and applied to screen the photoreactivity of the natural phenolic antioxidants caffeic acid, ferulic acid, p-coumaric acid, 3,4-dihydroxyphenylacetic acid (DOPAC), and rutin. The phototoxicity potential was determined by using a human keratinocyte cell line (HaCaT), based on the 3T3 Neutral Red Uptake phototoxicity test. Although all studied phenolic antioxidants absorbed UV/Vis radiation in the range of 290 to 700 nm, only DOPAC was able to generate singlet oxygen. The generation of reactive oxygen species is an early-stage chemical reaction as part of the phototoxicity mechanism. Yet, none of the studied compounds decreased the viability of keratinocytes after irradiation, leading to the conclusion that they do not have phototoxic potential. The data obtained with this work suggests that these compounds are safe when incorporated in cosmetic products.     

Radical Protection by Differently Composed Creams in the UV/VIS and IR Spectral Ranges

Modern sunscreens are well suited to provide sufficient protection in the UV range because the filter substances absorb or scatter UV radiation. Although up to 50% of radicals are formed in the visible and infrared spectral range during solar radiation protection strategies are not provided in this range. Previous investigations of commercially available products have shown that in addition to physical filters, antioxidants (AO) are necessary to provide protective effects in the infrared range by neutralizing already formed radicals. In this study, the efficacy of filter substances and AO to reduce radical formation in both spectral ranges was investigated after UV/VIS or IR irradiation. Optical properties and radical protection were determined for the investigated creams. It was found that organic UV filters lower radical formation in the UV/VIS range to 35% compared to untreated skin, independent of the presence of AO. Further reduction to 14% was reached by addition of 2% physical filters, whereas physical filters alone were ineffective in the UV/VIS range due to the low concentration. In contrast, this filter type reduced radical formation in the IR range significantly to 65%; similar effects were aroused after application of AO. Sunscreens which contain organic UV filters, physical filters and AO ensure protection in the complete solar spectrum.     

Isoflavonoid compounds from red clover (Trifolium pratense) protect from inflammation and immune suppression induced by UV radiation

Isoflavones derived from many edible plants have been reported to possess significant antioxidant, estrogenic and tyrosine kinase inhibitory activity. Genistein has been found previously to provide protection from oxidative damage induced by UV radiation both in vitro and following dietary administration. We have therefore examined the potential of a number of isoflavones from red clover (Trifolium pratense) and some metabolically related compounds to offer protection from UV irradiation in hairless mice by topical application after UV exposure. We show that whereas the primary isoflavones, daidzein, biochanin A and formononetin, were inactive, 20 microM lotions of genistein and the metabolites equol, isoequol and the related derivative dehydroequol had powerful potential to reduce the inflammatory edema reaction and the suppression of contact hypersensitivity induced by moderate doses of solar-simulated UV radiation. For equol the protection was concentration dependent and 5 microM equol markedly reduced the UV-induced inflammation but abrogated the UV-induced immunosuppression. Equol protected similarly from immunosuppression induced by the putative epidermal mediator, cis-urocanic acid (UCA), indicating a potential mechanism of action involving inactivation of this UV-photoproduct. Since immunosuppression induced by both UV radiation and by cis-UCA appears to be an oxidant-dependent response our observations support the actions of these topically applied isoflavones and their metabolites as antioxidants. They also indicate that lotions containing equol, unlike topical UV sunscreens, more readily protect the immune system from photosuppression than from the inflammation of the sunburn reaction, even when applied after exposure, and thus such compounds may have a future role as sun-protective cosmetic ingredients.     

Ultraviolet-induced responses in two species of climax tropical marine macrophytes.

In tropical regions nominal reductions in stratospheric ozone could be detrimental to marine organisms that live near their upper tolerance levels of ultraviolet (UV) radiation and temperature. Well-known plant responses to UV include inhibition of photosynthesis, reductions in chlorophyll content, morphological changes and production of UV absorbing compounds such as flavonoids. An assessment of the effects and responses of two tropical marine macrophytes to full solar radiation and solar radiation depleted of UV were conducted in southwestern Puerto Rico. Changes in concentration of photosynthetic and photoprotective pigments, and in leaf optical properties of the red mangrove Rhizophora mangle and the seagrass Thalassia testudinum, were evaluated in field exclusion experiments. Rhizophora mangle exposed to full solar radiation showed lower leaf reflectance and a shift of 5 nm in the inflection point of the red edge. Thalassia testudinum samples excluded from UV had significant increases in total chlorophyll and carotenoid concentrations. These marine macrophytes showed increments in their concentration of UV-B absorbing compounds with exposure to UV radiation. Results indicate that even minor increases in UV radiation at low latitudes could have significant effects on the pigment composition of these climax species.     

Solar PAR and UV radiation affects the physiology and morphology of the cyanobacterium Anabaena sp. PCC 7120

Solar UV radiation (280-400 nm) may affect morphology of cyanobacteria, however, little has been evidenced on this aspect while their physiological responses were examined. We investigated the impacts of solar PAR and UVR on the growth, photosynthetic performance and morphology of the cyanobacterium Anabaena sp. PCC7120 while it was grown under three different solar radiation treatments: exposures to (a) constant low PAR (photosynthetic active radiation, 400-700 nm), (b) natural levels of solar radiation with and (c) without UV radiation (290-400 nm). When the cells were exposed to solar PAR or PAR+UVR, the photochemical efficiency was reduced by about 40% and 90%, respectively, on day one and recovered faster under the treatment without UVR over the following days. Solar UVR inhibited the growth up to 40%, reduced trichome length by up to 49% and depressed the differentiation of heterocysts. Negligible concentrations of UV-absorbing compounds were found even in the presence of UVR. During the first 2 d of exposure to natural levels of PAR, carotenoid concentrations increased but no prolonged increase was evident. Heterocyst formation was enhanced under elevated PAR levels that stimulated quantum yield and growth after an initial inhibition. Higher concentrations of carotenoids and a twofold increase in the carotenoid to chlorophyll a ratio provided protection from the high levels of solar PAR. Under radiation treatments with UVR the relatively greater decrease in chlorophyll a concentrations compared with the increase in carotenoids was responsible for the higher carotenoid: chlorophyll a ratio. Heterocyst formation was disrupted in the presence of solar UVR. However, the longer term impact of heterocyst disruption to the survival of Anabaena sp. requires further study.     

The Photostabilizing Effect of Grape Seed Extract on Three Common Sunscreen Absorbers

The photostabilizing ability of grape seed extract on three common sunscreen absorbers, 2‐ethylhexyl‐p‐methoxycinnamate (EHMC), benzophenone‐3 (BP3) and tert‐butylmethoxy dibenzoylmethane (BMDBM), was investigated. Samples were exposed to simulated solar radiation and monitored by spectrophotometric and chromatographic methods. The chemical composition of the grape seed extract was determined by GC‐MS and HPLC‐MS, and the major secondary metabolites were found to be epicatechin and catechin. Exposure of the extract to UV radiation increased the UV absorption capacity of the extract. All sunscreens showed an improved photostability in the extract. The inherent photo‐instability of BMDBM when exposed to UV radiation was almost eliminated in the presence of grape seed extract. A mixture of all three sunscreens in the extract showed very high photostability and a red shift covering the entire UVB and UVA regions, thereby improving the broad‐spectrum protection. The incorporation of grape seed extract in sunscreen and other cosmetic formulations for topical application boosts photoprotection by stabilizing the UV filters and enhancing broad‐spectrum coverage. This in turn helps in reducing the amounts of absorbers and other additives incorporated in a sunscreen product and consequently lowers the risk of an unprecedented buildup of photoproducts whose toxicities are currently unknown.     

Effects of solar radiation on collagen and chitosan films

Photo-aging and photo-degradation are the deleterious effect of chronic exposure to sun light of many materials made of natural polymers. The resistance of the products on the action of solar radiation is very important for material scientists. The effect of solar radiation on two natural polymers: collagen and chitosan as well as collagen/chitosan blends in the form of thin films has been studied by UV-Vis and FTIR spectroscopy. It was found that UV-Vis spectra, which characterise collagen and collagen/chitosan films, were significantly altered by solar radiation. FTIR spectra of collagen and collagen/chitosan films showed that after solar irradiation the positions of amide A and amide I bands were shifted to lower wavenumbers. There was not any significant alteration of chitosan UV-Vis and FTIR spectra after solar radiation. In the condition of the experiment chitosan films were resistant to the action of solar radiation. The effect of solar UV radiation in comparison to artificial UV radiation has been discussed.     

The Use of Directional Reflectance Measurement for in vivo Assessment of Protective Properties of Cosmetics in the Infrared Radiation Range

Photoprotection of skin is now focused on UV radiation. The aim of this study was to evaluate in vivo cosmetic products in terms of protection against infrared radiation (IR) and propose a methodology for conducting such measurements. The directional reflectance (DR) of 12 UV filters, six care creams and two preparations containing fumed silica applied on the forearm of 36 volunteers was examined in six spectral bands for two angles of incidence. SOC‐410 Directional Hemispherical Reflectometer was used to measure DR. There is very little change in DR for all spectral bands for both incident angles for both UV filters, care creams and preparations containing fumed silica. For example, for 15% of fumed silica in glycerin for the spectral band of 0.9–1.1 μm and the incident angle of 20°, skin DR prior to application was 0.543; 5 min after application was 0.533 and 30 min after application was 0.559. Both UV filters, care creams and fumed silica do not protect skin against IR. The proposed method of in vivo measurements is superior to in vitro studies which have been conducted so far because it takes into account both the refractive index at the tissue/air interface and the absorption of IR by adipose tissue.     

Spectral properties of selected UV-blocking and UV-transmitting covering materials with application for production of high-value crops in high tunnels

The spectral properties of selected UV-blocking and UV-transmitting covering materials were characterized by means of a UV-VIS spectroradiometer or a UV-VIS spectrometer to provide researchers and growers with guidelines for selecting suitable materials for use in studying the effects of ambient solar UV radiation on the production of tomatoes and other high-value crops in high tunnels. A survey was made of a wide range of plastic covering materials to identify commercially available products that had the desired characteristics of transmitting high levels of photosynthetically active radiation and of being stable under ambient solar UV radiation. The study was focused on evaluating films that either blocked or transmitted UV wavelengths below 380 nm to determine comparative growth, yield and market quality and to provide a tool for integrated pest management. Based on this survey, two contrasting covering materials of similar thickness (0.152 mm) and durability (4-year polyethylene), one a UV-blocking film and the other a UV-transmitting film, were selected and used to cover two high tunnels at Beltsville, MD. Spectroradiometric measurements were made to determine comparative spectral irradiance in these two high tunnels covered with these materials and under ambient solar UV radiation. Comparative measurements were also made of selected glass and plastic materials that have been used in UV exclusion studies.     

Conidial pigmentation is important to tolerance against solar-simulated radiation in the entomopathogenic fungus Metarhizium anisopliae

The importance of conidial pigmentation to solar UV radiation tolerance in the entomopathogenic fungus Metarhizium anisopliae var. anisopliae, was estimated by comparing the effects of exposure to simulated solar UV radiation on the wild-type parent strain U.S. Department of Agriculture (USDA)-Agricultural Research Service (ARS) Collection of Entomopathogenic Fungal Cultures (ARSEF) 23, which has dark green conidia, and three groups of color mutants with yellow, purple and white conidia. The comparisons included inactivation levels and the kinetics of germination of conidia exposed or not exposed to simulated solar UV radiation. In addition to significantly inactivating the conidia of different mutants, exposure to radiation delayed for several hours the germination of surviving conidia of the wild type and all mutants. In general, mutants with white conidia were more sensitive to simulated solar UV radiation than mutants with purple conidia, which were more sensitive than mutants with yellow conidia, which in turn were more sensitive than the green wild strain. A significant variation in tolerance to simulated solar radiation was observed among mutants within each color group, particularly among mutants with yellow conidia. Revertants with green conidia, DWR 179 and DWR 176, were obtained from the very sensitive UV mutants DWR 148 (yellow conidia) and DWR 149 (purple conidia), respectively. These revertants had levels of tolerance to simulated solar UV radiation similar to those of the wild-type ARSEF 23. This observation is strong evidence of the importance of green conidial pigmentation for tolerance to simulated solar UV radiation, a factor that could be manipulated to produce M. anisopliae strains with more tolerance to solar UV radiation.     

Outdoor workers' acceptance of personal protective measures against solar ultraviolet radiation

The acceptance and usability of personal protection against solar UV radiation was evaluated in a field study with a group of tinsmiths in Austria. The personal protective measures (PPM) tested involved four categories: shirts, headwear, sunglasses and topically applied sunscreens; at least six different products per category were tested. Recommendations for the "ideal" shirt, headwear, pair of sunglasses and topical sunscreen are given based on data from questionnaires, i.e., from the point of view of the workers, independently from the actual physical level of protection (such as low transmittance or area of coverage) provided. It is argued that in practice it is important to consider the acceptance and usability of protective measures as well as the level of physical protection when providing PPM.     

Solar UV irradiation conditions on the surface of Mars

The UV radiation environment on planetary surfaces and within atmospheres is of importance in a wide range of scientific disciplines. Solar UV radiation is a driving force of chemical and organic evolution and serves also as a constraint in biological evolution. In this work we modeled the transmission of present and early solar UV radiation from 200 to 400 nm through the present-day and early (3.5 Gyr ago) Martian atmosphere for a variety of possible cases, including dust loading, observed and modeled O3 concentrations. The UV stress on microorganisms and/or molecules essential for life was estimated by using DNA damaging effects (specifically bacteriophage T7 killing and uracil dimerization) for various irradiation conditions on the present and ancient Martian surface. Our study suggests that the UV irradiance on the early Martian surface 3.5 Gyr ago may have been comparable with that of present-day Earth, and though the current Martian UV environment is still quite severe from a biological viewpoint, we show that substantial protection can still be afforded under dust and ice.     

Effects of solar ultraviolet radiation on photosynthesis of the marine red tide alga Heterosigma akashiwo (Raphidophyceae)

In order to assess the short- and long-term impacts of UV radiation (UVR, 280-400nm) on the red tide alga, Heterosigma akashiwo, we exposed the cells to three different solar radiation treatments (PAB: 280-700nm, PA: 320-700nm, P: 400-700nm) under both solar and artificial radiation. A significant decrease in the effective quantum yield (Y) during high irradiance periods (i.e., local noon) was observed, but the cells partially recovered during the evening hours. Exposure to high irradiances for 15, 30, and 60min under a solar simulator followed by the recovery (8h) under dark, 9 and 100micromolphotonsm(-2)s(-1) of PAR, highlighted the importance of the irradiance level during the recovery period. Regardless the radiation treatments, the highest recovery (both in rate and total Y) was found at a PAR irradiance of 9micromolphotonsm(-2)s(-1), while the lowest was observed at 100micromolphotonsm(-2)s(-1). In all experiments, PAR was responsible for most of the observed inhibition; nevertheless, the cells exposed only to PAR had the highest recovery in any condition, as compared to the other radiation treatments. In long-term experiments (10 days) using semi-continuous cultures, there was a significant increase of UV-absorbing compounds (UV(abc)) per cell from 1.2 to >4x10(-6)microgUV(abc)cell(-1) during the first 3-5 days of exposure to solar radiation. The highest concentration of UV(abc) was found in samples exposed in the PAB as compared to PA and P treatments. Growth rates (mu) mimic the behavior of UV-absorbing compounds, and during the first 5 days mu increased from <0.2 to ca. 0.8, and stayed relatively constant at this value during the rest of the experiment. The inhibition of the Y decreased with increasing acclimation of cells. All our data indicates that H. akashiwo is a sensitive species, but was able acclimate relatively fast (3-5 days) synthesizing UV-absorbing compounds and thus reducing any impact either on photosystem II or on growth.     

The Angular Distribution of Solar Ultraviolet, Visible and Near-Infrared Radiation from Cloudless Skies

Abstract— The amount of solar radiation intercepted by an object depends on the orientation of the object with respect to the sun and the angular distribution of the diffuse component of solar radiation, which is commonly considered to be approximately isotropic. The angular distribution of the diffuse UV, visible and near-infrared insolation was measured at several solar zenith angles between 32° and 68° under cloudless skies at Lauder, New Zealand (45S), and shown to be anisotropic. The diffuse solar UV radiation increases markedly with solar elevation and is a large proportion of the total UV irradiance. The diffuse visible light and infrared radiation are small components of the total irradiance and almost independent of solar elevation. The angular distribution of erythemal UV radiation was tabulated and is available on request.     

Synthesis and antioxidant activity of modified sterically hindered phenols

Salicylaldehyde derivatives containing sterically hindered phenol fragments were synthesized. The compounds obtained are capable to offer protection against UV radiation.     

Leaves of Citrus aurantifolia exhibit a different sensibility to solar UV-B radiation according to development stage in relation to photosynthetic pigments and UV-B absorbing compounds production

Plants of Citrus aurantifolia grown in a greenhouse without solar UV radiation (UVR) were transferred outdoors to evaluate the effect of solar UV-B radiation (UVBR, 280–315 nm) in prior-developed leaves, constituted by apical bud and those fully expanded before being taken outdoors, and post-developed leaves, formed by those expanded outdoors. Results demonstrated that over a 40 d outdoor period leaf chlorophyll content and distribution pattern were different with and without solar UVBR. Chlorophyll a, chlorophyll b and total chlorophyll contents in both treatments were higher in prior-developed leaves than in post-developed ones. However, highest values were observed in prior-developed leaves under solar UVBR, whereas in post-developed leaves an opposite trend was observed. Carotenoids content in prior-developed leaves was higher with solar UVBR, whereas in post-developed leaves there were no significant differences in both with and without solar UVBR. In addition, prior-developed leaves under solar UVBR accumulated flavonoids, but not anthocyanins. Growth parameters (e.g. DW, DW/FW ratio, LMA, plant height, length and width of foliar lamina) did not show significant differences between plants grown with and without solar UVBR. Thus, our results demonstrated that C. aurantifolia leaves exhibited a different sensibility to solar UVBR according to development stage in relation to photosynthetic pigments and UV-B absorbing compounds production. In addition, the solar UVBR was not necessary as inductor of photosynthetic protection mechanisms in a short-time growth period. On the other hand, our results also demonstrated that solar UVBR acted as an effective feeding deterrent against citrus leafminer.     

Standard ultraviolet daylight for nonextreme exposure conditions

The skin is exposed to ultraviolet radiation (UVR) from natural or artificial sources on a daily basis. The effects of chronic low dose exposure merit investigation, even when these effects are neither conspicuous nor clinically assessable. The purpose of the present study was to define a relative spectral UV irradiance that is representative of frequent nonextreme sun exposure conditions and therefore more appropriate for studies of the long-term and daily effects of solar UV on the skin. Solar spectral UV irradiance values were calculated for different dates and locations by using a radiative transfer model. The spectral irradiance values obtained when the solar elevation is lower than 45 degrees were averaged. An important feature is the dUVA (320-400 nm) to dUVB (290-320 nm) irradiance values ratio, which was found to be 27.3 for the overall average. When the months corresponding to extreme irradiance values (low or high) were excluded from the calculations, the dUVA to dUVB ratio ranged from 27.2 to 27.5. The mean spectral irradiance of the model presented here represents environmental UV exposure conditions and can be used both as a standard to investigate the biological effects of a nonextreme UVR and to assess the effectiveness of products for daily skin protection.     

Ultraviolet Spectral Energy Differences Affect the Ability of Sunscreen Lotions to Prevent Ultraviolet-Radiation-Induced Immunosuppression*

Acute exposure to UV radiation causes immunosuppression of contact hypersensitivity (CH) responses. Past studies conducted with unfiltered sunlamps emitting nonsolar spectrum UV power (wavelengths below 295 nm) or using excessive UV doses have suggested sunscreens may not prevent UV-induced immunosuppression in mice. This study was thus designed to evaluate critically the effects of different UV energy spectra on the immune protection capacity of sunscreen lotions. Minimum immune suppression doses (MISD), i.e. the lowest UV dose to cause~50% suppression of the CH response to dinitrofluorobenzene in C3H mice, were established for three artificial UV sources. The MISD for each UV source was 0.25 kJ/m² for unfiltered FS20 sunlamps (FS), 0.90 kJ/m² for Kodacel-filtered FS20 sunlamps (KFS), which do not emit UV power at wavelengths <290 nm, and 1.35 kJ/m² for a 1000 W filtered xenon arc lamp solar simulator. Using MISD as baseline, sunscreens with labeled sun protection factors (SPF) of 4, 8, 15 and 30 were tested with each UV source to establish their relative immune protection factors. The immune protection factor of each sunscreen exceeded its labeled SPF in tests conducted with the solar simulator, which has a UV power spectrum (295–400 nm) similar to that of sunlight. Conversely, sunscreen immune protection factors were significantly less than the labeled SPF in tests conducted with FS and KFS. Comparison of the immunosuppression effectiveness spectra showed that relatively small amounts of nonsolar spectrum UV energy, i.e. UVC (200–290 nm) and/or shorter wavelength UVB (between 290 and 295 nm), produced by FS and KFS contributes significantly to the induction of immunosuppression. For example, 36.3% and 3.5% of the total immunosuppressive UV energy from FS and KFS, respectively, lies below 295 nm. Sunscreen absorption spectra showed that transmission of immunosuppressive UV energy below 295 nm for FS was at least eight-fold higher than that for KFS. Compared to the solar simulator UV spectrum the transmission of nonsolar immunosuppressive UV energy through sunscreens was >15-fold higher for FS and ≥1.5-fold higher for KFS. These data demonstrate that relevant evaluations of sunscreen immune protection can only be obtained when tests are conducted with UV sources that produce UV power spectra similar to that of sunlight and UV doses are employed that are based on established MISD.