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Evaluation of the effects of cosmetic or dermo-pharmaceutical products on cutaneous energy metabolism using the Episkin model of reconstructed epidermis.

This study was implemented to test the Episkin model of reconstructed epidermis in the evaluation of the efficacy of cosmetic or dermopharmaceutical products on cutaneous energy metabolism. The energy metabolism is evaluated by measuring the concentration of intracellular ATP by a method using an ultrasensitive bioluminescent reaction. The work presented compares results obtained in reconstructed epithelium and monolayer primary cultures of human keratinocytes. After application of a hydrosoluble product, the increase in intracellular ATP is identical in a monolayer culture of keratinocytes (+239 +/- 18% versus control) and in Episkin (+248 +/- 21% versus control). An emulsion was also tested on the two models. It is only possible to test the emulsion at a dilution of under 0.05% on a keratinocyte culture, and this means that the real efficacy of the product is underestimated (+145 +/- 18% versus control). The three-dimensional model enables the application of the undiluted emulsion, and the results show an increase in intracellular ATP of +420 +/- 80% versus control: products in final formulation can be tested in normal conditions of use.

Adenosine Triphosphate↗

An impedimetric method for rapid screening of cosmetic preservatives.

An efficient impedance method was developed for rapid evaluation of cosmetic preservatives. The method used decimal reduction time or D-value to assess preservative efficacies. The D-value, which was calculated from the plot of Log CFU ml-1 versus time by linear regression analysis, could be obtained within 48 h. Thus, the time required for the challenge test was reduced from 4-8 weeks with the standard procedures (eg US Pharmacopeia), to 2 days with the current method. A calibration curve (r = -0.95) was established by plotting the Log CFU ml-1 versus capacitance detection time (DT) of 108 samples. With the calibration, CFU can be estimated directly from the impedance test without plating. Two commercial biocides and several other chemicals were evaluated in a shampoo by the impedance procedure against Pseudomonas aeruginosa. The D-values obtained from the impedance test were not significantly different from those produced by the conventional plate count method. The technique was found to be particularly useful when screening a large number of compounds to find novel preservatives and synergistic preservative combinations.

Cosmetics↗

Possible link between anaphylactoid reactions to anaesthetics and chemicals in cosmetics and biocides.

Binding-inhibition studies involving human IgE antibody to suxamethonium, SUX (from a patient with a near-fatal reaction), were performed with five different quaternary ammonium compounds (QAC): three common ingredients of cosmetics and two commonly used disinfectants, cetrimide and benzalkonium. All the five QAC showed immunological cross-reaction with SUX. In addition, at high concentrations, they released histamine from basophils of normal subjects. Thus, QAC may be both sensitizers and histamine releasers.

Anaphylaxis↗

Relationship between tyrosinase inhibitory action and oxidation-reduction potential of cosmetic whitening ingredients and phenol derivatives.

The oxidation-reduction potentials of cosmetic raw materials, showing tyrosinase inhibitory action, and phenolic compounds structurally similar to L-tyrosine were determined by cyclic voltammetry. The voltammograms obtained could be classified into 4 patterns (patterns 1-4). Pattern 1, characterized by oxidation and reduction peaks as a pair, was observed with catechol, hydroquinone or phenol, and pattern 2 exhibiting another oxidation peak in addition to oxidation and reduction peaks as a pair was found with arbutin, kojic acid, resorcinol, methyl p-hydroxybenzoate and L-tyrosine as the substrate of tyrosinase. Pattern 3 with an independent oxidation peak only was expressed by L-ascorbic acid, and pattern 4 with a reduction peak only at high potentials, by hinokitiol. The tyrosinase inhibitory activity of these compounds was also evaluated using the 50% inhibitory concentration (IC50) and the inhibition constant (Ki) as parameters. Hinokitiol, classified as pattern 4, showed the highest inhibitory activity (lowest IC50 and Ki). Hydroquinone showing the second highest activity belonged to pattern 1, which also included compounds showing no inhibition of tyrosinase activity. The inhibitory activity of compounds exhibiting pattern 2 was relatively low with Ki values being in the order of 10(-4) M. Although there was no consistent relationship between oxidation-reduction potentials and tyrosinase inhibitory action, the voltammetry data can be used as an additional index to establish the relationship between the structure and the tyrosine inhibitory activity.

Cosmetics↗

New tool for epidermal and cosmetic formulation studies by attenuated total-reflection spectroscopy using a flexible mid-infrared fiber probe.

The potential of mid-infrared attenuated total-reflection spectroscopy for dermatology studies has been increased by the development of a flexible fiber-optic probe from silver halide material. Such a tool significantly eases epidermal surface characterization. Spectra of human forearm skin and of bovine udder skin (BUS-model), which is used as a substitute for in-vivo tests on man, were compared, and as an example low concentrations of vitamin E acetate (alpha-tocopherol acetate) in the stratum corneum were measured after topical skin application. Depth profiling was enabled by repeated steps of tape stripping and subsequent skin measurement. The infrared methodology is rapid and can be applied to small areas of skin. It is also sufficiently sensitive for analysis of a large variety of cosmetic formulations and dermatopharmaca. Discrimination of vehicle formulations is possible because of the large information content of mid-infrared spectra.

Adult↗

Mechanisms of cytotoxicity by cosmetic ingredients in sea urchin eggs.

The acute cytotoxicities of four cosmetic ingredients: a preservative, imidazolidinylurea (IU) and three mild surfactants, cocamido propyl hydroxy sultaine (CAS), magnesium laureth sulfate (Mg LES), and decyl glucoside (APG) were studied using sea urchin eggs. The cellular targets of these compounds were identified by studying the effects on calcium homeostasis, intracellular pH, sodium and potassium contents, protein and DNA synthesis, and protein phosphorylation. These compounds inhibited the first cleavage of sea urchin eggs in a dose-dependent fashion with half maximal doses (IC50) from 30 microg/ml for Mg LES, 60 microg/ml for IU, 83 microg/ml for CAS, to above 400 microg/ml for APG. The time at which a compound showed the greatest toxicity to the cell cycle was definable for APG (between 20 and 50 min postfertilization) and IU (from fertilization to 50 min later); the other compounds being toxic throughout division. Compounds exhibited toxicity to a wide range of cellular targets. IU, the least toxic, mainly operates through inhibition of protein and DNA syntheses. CAS and Mg LES produced nonspecific cytotoxicity related to alterations of membrane and endomembrane permeabilities resulting in ionic disequilibrium (Na+, K+, Ca2+) and inhibition of intracellular storage of Ca2+. The APG effect mainly involved intracellular pH and DNA synthesis, a hypothesis suggested by the narrow postfertilization period of maximal toxicity.

Animals↗

Evaluation of the potential carcinogenicity of paraffins for medicinal and cosmetic uses--determination of polycyclic aromatic hydrocarbons.

A previously developed method for the determination of polycyclic aromatic hydrocarbons (PAH) in purified petroleum products was applied to some fully-refined paraffin samples for cosmetic and medicinal uses of Italian Pharmacopoeia grade materials. The analytical procedure used provided identification and quantitative assay for ng/g amounts of two well-known carcinogenic PAH, namely benzo[a]pyrene and benzo[b]fluroanthene, together with five other none-carcinogenic PAH. The method is sensitive and reliable for routine analysis of PAH content in these paraffins and is compared with very rapid pharmocopoeial tests based on different criteria of quality. The data obtained showed a lack in agreement between PAH content in paraffin samples and the results of pharmacopoeial tests.

Carcinogens↗

A review of the genotoxicity of food, drug and cosmetic colours and other azo, triphenylmethane and xanthene dyes.

The genetic toxicology of the major dyestuffs used in foods, drugs and cosmetics has been reviewed. Published data for azo, triphenylmethane and xanthene dyes from short-term assays for muta-carcinogenicity have been summarized and discussed according to usage, current and previous worldwide legislative status. Certain other synthetic food dyes, commercial mixtures, natural and polymeric colourants as well as a section on aminoazobenzene and its derivatives have been included. Genotoxicity has been discussed with reference to structural chemistry, levels of exposure, absorption and metabolism and to epidemiological information. The extent of agreement between data from different tests and correlations with animal cancer assays have been considered. Synthetic dyes from the 3 major structural classes exhibit genotoxicity, whilst only 2 natural colours have proved active. Activity may be due to the presence of certain functional groups, notably nitro- and amino-substituents which are metabolized to ultimate electrophiles that may be stabilized by electronic interaction with aryl rings. Metabolic processes such as azo-reduction may be activating or detoxifying. the low but significant correlation between animal carcinogenicity and short-term test data may be increased with further screening, especially involving chromosome assays. It is suggested that a human cancer hazard may exist where significant quantities of finished benzidine dye samples are handled. Such risks from exposures to other colours and the possibility of human germ-line mutation induction by dyestuffs cannot be meaningfully assessed.

Animals↗

Mutagenicity of cosmetics ingredients licensed by the European Communities.

As part of our investigation into mutagenic effects of environmental compounds, we studied chemicals allowed as ingredients of cosmetics according to the guidelines of the Council of the European Communities (27 July 1976). We used three systems, the Salmonella/microsome test, the Basc test on Drosophila and the micronucleus test on mouse bone marrow. Of the 31 chemicals tested, 15 were mutagenic in the Ames test; and of these, 5 were also mutagenic in the Basc test and 2 in the micronucleus test.

Animals↗

Evaluation by electron microscopy techniques of asbestos contamination in industrial, cosmetic, and pharmaceutical talcs.

Talc powders from national and international markets were analyzed in order to assess their fiber contents and the proportion of asbestos in the fibrous material. Samples of talc powders used as excipients in pharmaceutical and cosmetic preparations demonstrated fiber contents up to 30% of total particles. About a half of the talc powders revealed the presence of asbestos: in five samples chrysotile (a serpentine asbestos) was present, in the other ones tremolite and anthophyllite (an amphibole asbestos). The amounts of asbestos vary up to 90% in the different samples of the fibrous fraction. About 75% of observed asbestos fibers were thinner than 0.4 micron, i.e., below the resolving power of light microscopy which until now was the most utilized technique for evaluating the environmental pollution due to asbestos.

Asbestos↗

Long-chain N-nitroso-N-methylalkylamines in commercial cosmetics, light-duty dishwashing liquids and household cleaning preparations.

A method for the determination of long-chain N-nitroso-N-methylalkylamines in a range of commercial cosmetic and cleaning emulsions is described. The clean-up consists of solid-phase extraction and column chromatography. N-Nitroso-N-methyldodecylamine, N-nitroso-N-methyltetradecylamine and N-nitroso-N-methylhexadecylamine are detected by gas-liquid chromatography using a Thermal Energy Analyzer. N-Nitroso-N-ethyldodecylamine is used as an internal standard. Analysis of 86 products produced and marketed in the FRG did not indicate significant contamination. In contrast, of 26 consumer products from the USA, three (one hair-care product and two dishwashing liquids), which contained fatty amine oxides, were contaminated with substantial amounts of N-nitroso-N-methylalkylamines.

Chromatography, Gas↗

Aflatoxins in cosmetics containing substrates for aflatoxin-producing fungi.

29 random samples of eight different products of skin cleansing creams or powders and skin peeling products (including practically all products of this type available on the German market), containing substrates for aflatoxin-producing fungi or plant parts directly connected with such substrates, such as almond bran, peach bran, apricot seeds, wheat bran, horse chestnut and cob were analysed for aflatoxin B1, B2, G1 and G2. In 11 (38%) of the samples aflatoxins were not detectable ( < 0.1 micrograms/kg); 18 (62%) of the samples contained from 0.4 to 78.5 micrograms aflatoxin/kg. The majority of the positive samples contained all four types of aflatoxin. Aflatoxins can be absorbed through the skin. To what extent this happens with these products and the resulting toxicological significance is difficult to estimate. Nevertheless, potent carcinogens such as aflatoxins should be completely absent from cosmetics; this can be achieved by careful quality control of the raw materials.

Aflatoxins↗

The safety and regulatory status of food, drug and cosmetics colour additives exempt from certification.

Colour additives exempt from certification (also known as 'natural colour additives') are commonly used in the United States to colour foods, drugs and cosmetics. The US Food and Drug Administration established regulations governing the use of these colour additives, and the labelling of the products that contain them. The safety of these colour additives has been demonstrated by safety testing programs that have yielded a significant amount of toxicology data, and also by a long and well documented history of safe use in the United States and elsewhere. In this report we review the data supporting the safety of the colour additives that are exempt from certification and approved for use in the USA, and conclude that the safety of these colour additives is well supported by the available data. Relevant aspects of the regulation of these colour additives are also described.

Cosmetics↗

Plants used in phytotherapy, cosmetics and dyeing in the Pramanda district (Epirus, North-West Greece).

Thirty plants used in phytotherapy, cosmetics and dyeing in the Pramanda district (North-West Greece) are reported. For many plants (myrtle, phlomis, daphne, etc.) the uses are localized and not noted for other phytotherapies; some uses coincide with those existing in other zones of the Mediterranean (mint, thyme, red juniper, Hypericum, etc.). The plants used are the most common and most easily found in this area.

Coloring Agents↗

Analysis of aliphatic dicarboxylic acids in pharmaceuticals and cosmetics by liquid chromatography (HPLC) with fluorescence detection.

2-Bromoacetyl-6-methoxynaphthalene has been found to be a useful prechromatographic fluorescent labelling reagent for the analysis of dicarboxylic acids. The derivatization reaction of azelaic acid and meglutol with this reagent yieded stable and highly fluorescent diesters which could be analysed by reversed-phase HPLC with fluorescence detection. According to the nature of the sample, the derivatization reaction could be carried out in acetonitrile or in an aqueous micellar system. The proposed methods proved to be suitable for the quality control of various complex pharmaceutical and cosmetic formulations of the azelaic acid and meglutol.

Chromatography, High Pressure Liquid↗

Determination of antioxidants in cosmetics by micellar electrokinetic capillary chromatography with electrochemical detection.

A new and efficient method for the determination of antioxidants [Propyl gallate (PG), tert-butylhydroquinone (TBHQ), butylated hydroxyanisole (BHA), and butylated hydroxytoluene (BHT)] in cosmetics has been established by using micellar electrokinetic capillary chromatography with electrochemical detection (MECC-ED). Under the optimum conditions of the method, all analytes were successfully separated within 13 min at the separation voltage of 18 kV in a 20 mmol/L borate running buffer (pH 7.4) containing 25 mmol/L sodium dodecyl sulfate. The excellent linearity was obtained in the concentration range from 5.0 x 10(-4) to 2.0 x 10(-6) mol/L and the detection limits (S/N = 3) of PG, TBHQ, BHA, and BHT range from 3 x 10(-7) to 3 x 10(-6) mol/L.

Antioxidants↗

Determination of fragrance allergens in cosmetics by size-exclusion chromatography followed by gas chromatography-mass spectrometry.

A method using size-exclusion chromatography (SEC) combined with gas chromatography-mass spectrometry (GC-MS) was developed for the quantitation of 24 restricted allergenic fragrance compounds in cosmetic samples. To achieve reproducible results fragrance calibration has to be performed with propyl acetate as a solvent containing a constant proportion of matrix components. With the exception of hydroxycitronellal (66+/-5%) all compounds showed good recovery rates in the range of 90-120%. The mean accuracy (relative error) was 1+/-10% for all 24 compounds in five spiked creams (10 mg/kg per allergen) and 8+/-34% in a reference sample (4-15 mg/kg). The biggest benefit compared to other methods is the flexible clean up with SEC which allows the determination of a large range of compounds in difficult matrices with GC-MS.

Allergens↗