Evaluation for collagen products for cosmetic application

The assessment of ocular irritation potential is an important part of safety testing for cosmetic and consumer products. The purpose of this investigation was to examine ocular irritancy levels elicited in humans by various categories of a specific class of cosmetic and consumer products that have a potential to enter the eye inadvertently during use. Test materials assessed belonged to one of seven categories, which included liquid make-up, shampoo, baby wash, mascara, eye make-up remover, powder eye shadow, and facial cleanser. These test materials were evaluated by human ocular instillation, followed by examinations, for which subjective perceptions of irritation were recorded, and component areas of ocular tissues were individually examined for inflammation and for the area and density of fluorescein staining patterns at 30 s and at 5, 15, 60, and 120 min post-instillation. Subjective and objective ocular irritation scores of 410 eyes were analyzed by product classification. Average score levels were determined for subjective responses, inflammation, and fluorescein staining patterns. This investigation determined that irritation levels of the evaluated test materials varied markedly with respect to product category, type of ocular irritation, and ocular tissue, demonstrating that these factors are important considerations for the prediction of the ocular irritancy of a test material.     

Hydrolysed ginseng-saponin quaternary: a novel conditioning agent for hair care products

A new quaternary ammonium compound, hydrolysed ginseng-saponin quaternary (HGSQ), from Korean ginseng saponin and 2,3-epoxypropyltrimethyl ammonium chloride, has been developed as a conditioning agent for hair care products. This structure has a hydrophobic group from the aglycone of ginseng saponin which is biologically active and considered as the most important component of Korean ginseng. Its properties of surface tension, conductivity, critical micelle concentration (CMC), eye irritation, sorption onto hair, tensile strength for 20% extension and moisture retention effect were studied. Its cationic character allows the molecule to be more substantive than ginseng saponin. HGSQ had good physical properties and was safe enough as a cosmetic raw material. Also half-head tests of HGSQ-containing shampoo were carried out to evaluate the conditioning effects in shampoos. HGSQ showed good conditioning properties in a shampoo.     

发用类、肤用清洁类化妆品的急性皮肤和眼刺激损害特点

分析发用类、肤用清洁类化妆品的急性皮肤刺激和急性眼刺激损害特点,了解其卫生安全状况。于2012年采用《化妆品卫生规范》(2007年版)中的急性皮肤刺激性实验和急性眼刺激性实验对发用类、肤用清洁类共计158种化妆品样品进行测试。两类受试化妆品显示出不同程度的急性皮肤刺激性和急性眼刺激性损害效应。发用类化妆品在实验中基本无皮肤刺激性损伤;肤用清洁类化妆品在实验中有少量的皮肤刺激性损伤,但大都为轻刺激性(31.0%),只有个别(3.8%)出现了皮肤红斑水肿的中刺激性损伤。发用类、肤用清洁类化妆品在急性眼刺激性实验中检出一定比例具有微刺激性及以上刺激性的样品,但引起的眼损伤都能够在观察期限内恢复。本次调查的所有化妆品中,洁面类、洗发类化妆品普遍具有急性眼刺激性和急性皮肤刺激性损害,需加强对化妆品的监督管理,进一步提升化妆品卫生安全水平,保障化妆品使用安全。     

Assessment of surfactant cytotoxicity: comparison with the Draize eye test

We have compared the in vitro cellular toxicity and the in vivo ocular irritation potency of 16 surfactants (7 non ionics, 3 anionics, 2 amphoterics, 4 cationics) ranking from very weakly irritant to strongly irritant. In vitro , the cellular toxicity was estimated on Chinese hamster lung fibroblasts (V79) using a cell mortality test and a cell growth inhibition test. For each surfactant, a lethal concentration 50% without foetal calf serum (LC.50–0) and with 10% foetal calf serum (LC50–I0), as well as the concentration required to reduce 50% of the growth (CI.50) were determined. In vivo , each surfactant was applied directly to the cornea of six albino rabbits. The maximal ocular irritation score (I0 max) and the ocular irritation score obtained seven days later (I0 J7) were collected. Comparison of in vitro results with those obtained in vivo showed good correlations, particularly when I0 max and the difference (LC.50–10 - LC50–0) were considered ( r = 0.845, P<0.001 ). These results suggest that the use of cell culture tests as pre-screening systems to appreciate eye irritation potency of surfactants could be a reliable alternative method in order to reduce the use of the Draize rabbit eye test. They can provide a better knowledge of the irritative process induced by surfactants (cellular toxicity and protein interaction potency).     

Targeted delivery of salicylic acid from acne treatment products into and through skin: role of solution and ingredient properties and relationships to irritation

Salicylic acid (SA) is a beta‐hydroxy acid and has multifunctional uses in the treatment of various diseases in skin such as acne, psoriasis, and photoaging. One problem often cited as associated with salicylic acid is that it can be quite irritating at pH 3–4, where it exhibits the highest activity in the treatment of skin diseases. We have identified strategies to control the irritation potential of salicylic acid formulations and have focused on hydroalcoholic solutions used in acne wipes. One strategy is to control the penetration of SA into the skin. Penetration of the drug into various layers of skin, i.e. epidermis, dermis, and receptor fluid, was measured using a modified Franz in vitro diffusion method after various exposure times up to 24 h. A polyurethane polymer (polyolprepolymer‐15) was found to be an effective agent in controlling delivery of SA. In a dose‐dependent fashion it targeted delivery of more SA to the epidermis as compared to penetration through the skin into the receptor fluid. It also reduced the rapid rate of permeation of a large dose of SA through the skin in the first few hours of exposure. A second strategy that proved successful was incorporation of known mild nonionic surfactants like isoceteth‐20. These surfactants cleanse the skin, yet due to their inherent mildness (because of their reduced critical micelle concentration and monomer concentration), keep the barrier intact. Also, they reduce the rate of salicylic acid penetration, presumably through micellar entrapment (either in solution or on the skin surface after the alcohol evaporates). Cumulative irritation studies showed that targeting delivery of SA to the epidermis and reducing the rapid early rate of penetration of large amounts of drug through the skin resulted in a reduced irritation potential. In vivo irritation studies also showed that the surfactant system is the most important factor controlling irritancy. SA delivery is secondary, as formulations with less SA content reduced the rate of delivery to the receptor and yet were some of the most irritating formulations tested, presumably due to the action of the specific anionic surfactant on the barrier. Alcohol content also did not appreciably affect irritation and SA delivery; formulations with considerably low alcohol content but containing anionic versus nonionic surfactant systems exhibited considerably higher irritancy. Thus the surfactant type was again the predominant factor in those studies, although arguably alcohol plays some role (solubilization of SA). Results showed that both polymers and mild surfactants work in concert to provide the optimal formulation benefits of targeted delivery and reduced irritation. Synergistic relationships among hydroalcoholic formulation components will be discussed along with the mechanisms likely involved in controlling delivery of SA to skin.     

Efficacy of a piroctone olamine/climbazol shampoo in comparison with a zinc pyrithione shampoo in subjects with moderate to severe dandruff

Dandruff is a chronic scalp disorder characterized by scaling and itching. A successful anti‐dandruff shampoo not only has to provide superior anti‐dandruff relief to ensure patient compliance. It also needs to offer excellent cosmetic and hair conditioning benefits at the same time. In this study, the efficacy of a shampoo containing 0.5% piroctone olamine and 0.45% climbazole (shampoo 1) was compared with a widely available commercial shampoo containing 1% zinc pyrithione (shampoo 2). In vitro studies investigating the anti‐mycotic efficacy of a combination of 0.5% piroctone olamine and 0.45% climbazole as well as 1% zinc pyrithione were performed. To study substantivity, pig skin punches were used as a model system and a test of wet combability was performed to characterize combing ease. In vivo home‐in‐use studies were carried out to determine the efficacy of both shampoos to improve scalp condition and reduce itching in subjects suffering from moderate to severe dandruff. Results demonstrated a comparable anti‐fungal effectiveness for 0.5% piroctone olamine plus 0.45% climbazole and 1% zinc pyrithione, respectively. Shampoo 1 showed a significantly higher anti‐mycotics substantivity compared to shampoo 2. After treatment with shampoo 1, the wet combing force was significantly reduced compared with shampoo 2, suggesting a better combability following the use of shampoo 1. In an in vivo split head design study, shampoo 1 was shown to be equally effective in reducing the amount of dandruff on the scalp compared with shampoo 2. The approval rate of volunteers regarding the question ‘The use of this shampoo decreases the itching of my scalp?’ after a 4‐week treatment with shampoo 1 equaled 90%. Overall, the shampoo formulation with 0.5% piroctone olamine and 0.45% climbazole effectively reduces the amount of dandruff and, at the same time, provides hair conditioning advantages.     

Study of surfactant-skin interactions by skin impedance measurements

The stratum corneum (SC) plays a very critical physiological role as skin barrier in regulating water loss through the skin and protects the body from a wide range of physical and chemical exogenous insults. Surfactant-containing formulations can induce skin damage and irritation owing to surfactant absorption and penetration. It is generally accepted that reduction in skin barrier properties occurs only after surfactants have penetrated/permeated into the skin barrier. To mitigate the harshness of surfactant-based cleansing products, penetration/permeation of surfactants should be reduced. Skin impedance measurements have been taken in vitro on porcine skin using vertical Franz diffusion cells to investigate the impact of surfactants, temperature and pH on skin barrier integrity. These skin impedance results demonstrate excellent correlation with other published methods for assessing skin damage and irritation from different surfactant chemistry, concentration, pH, time of exposure and temperature. This study demonstrates that skin impedance can be utilized as a routine approach to screen surfactant-containing formulations for their propensity to compromise the skin barrier and hence likely lead to skin irritation.     

In vivo procedures for assessment of hair greasiness

Greasy hair is a common problem in Europe. The first step in developing anti-grease hair products must be to establish a sensitive protocol for measuring any changes in perceived hair greasiness.
Sensitive clinical trials and in vivo evaluation methods of determining hair greasiness have been developed that show significant differences in the perceived hair greasiness following the use of different shampoos.
Products tested in two clinical trials for efficacy as anti-grease shampoos were an anti-grease shampoo containing 2% 3,4-thiolanediol as the active ingredient, and a baby shampoo based on mild surfactants. The same placebo shampoo based on ether sulphates used in typical European shampoo formulations was used in both tests. A shampoo containing 2% zinc pyrithione was included in the trials as a control, since there were indications that this would increase the amount of hair greasiness.
Neither of the test products were shown to be effective anti-grease shampoos. The shampoo containing 2% zinc pyrithione was shown to produce significantly more greasiness than both the placebo and the test shampoos.
Since significant differences could be shown between the different shampoos, the protocol and in vivo evaluation techniques used in these clinical trials are considered to be validated.
Méthodes in vivo d'évaluation du graissage des cheveux     

Skin improvement and stability of Echinacea purpurea dermatological formulations

Echinacea purpurea contains many beneficial constituents for protection of skin from oxidative stress and for improving hydration of skin. This study aimed to investigate the stability and dermatological efficacy of E. purpurea cream and gel. Echinacea purpurea extract was incorporated into suitable cream and gel bases. Stability of the extract in the formulations was investigated by determining its residual total phenolic content and antioxidant activity after storage at 4°C, 30°C and 40°C for 6 months. The effect of those formulations on skin irritation, hydration level and wrinkle reduction was evaluated in 10 healthy volunteers, aged 25–40 years. The shelf lives of E. purpurea cream and gel in terms of total phenolic content and antioxidant activity were only 2 and 4 months respectively at 4°C and could be extended up to 7 months by incorporation of α‐tocopherol or disodium editate. The corneometer hydration indices increased up to 10.6 AU and 11.4 AU, and the wrinkles decreased 9.47% and 14.92% because of the application of E. purpurea cream and gel for 1 month. Both formulations showed no irritation to skin. Echinacea purpurea cream and gel developed in this study were effective in improving skin hydration and reducing wrinkle, but showed low storage stability.     

Degradation kinetics of rebaudioside A in various buffer solutions

Rebaudioside A is a noncaloric high‐intensity sweetener extracted from Stevia rebaudiana. For it to be used in foods, rebaudioside A needs to be stable during processing and storage. This study evaluated the kinetics of rebaudioside A degradation in 0.02 and 0.1 m citrate and phosphate buffer solutions at pH 3, 5 and 7 stored at 20, 30 and 40 °C for 9 months. At pH 3 and temperatures less than 30 °C, degradation was slow (i.e. less than 6% loss in 6 months) and not affected by buffer type or buffer concentration. At pH 7, the degradation of rebaudioside A increased dramatically in phosphate buffer (losses of up to 25% in 6 months at 30 °C), while remaining relatively slow in citrate buffer (less than 5% loss). The degradation rates at pH 7 increased 2.6 times as the concentration of phosphate buffer increased. For optimum stability of beverages containing rebaudioside A, formulations with lower concentrations of citrate buffer stored at lower temperatures are recommended.     

Rheological characterization of hair shampoo in the presence of dead sea salt

In Jordan, a growing industry has been established to produce different types of Dead Sea (DS) cosmetics that have DS salt (contains mainly NaCl, KCl, and MgCl(2)) in their formulas. In this work, the effect of DS salt on the rheology of hair shampoo containing the sodium lauryl ether sulfate as a main active matter was studied. The effects of DS salt and active matter concentration, and the temperature and time of salt mixing, on the rheological properties of hair shampoo were investigated. The salt-free shampoo showed a Newtonian behavior at 'low active matter' (LAM) and shear thinning at 'high active matter' (HAM). The presence of DS salt changed the rheological behavior of LAM shampoo from Newtonian (for the salt-free shampoo) to shear thinning. On the other hand, the behavior of HAM shampoo switched from shear thinning to Newtonian behavior in the presence of high concentration of DS salt. The addition of DS salt increased the apparent viscosity of shampoo to reach a maximum value that corresponded to a salt concentration of 1.5 wt.%. Further addition of DS salt led to a decrease in the shampoo viscosity to reach a value less than that of the salt-free sample at high salt concentration. Changing the mixing temperature (25-45 degrees C) and mixing time (15-120 min) of DS salt with shampoo has no significant influence on the rheological behavior. However, the mixing process increased the apparent viscosity of salt-free shampoo. The power law model fitted well the flow curves of hair shampoo with and without DS salt.     

Rheology of Dead Sea shampoo containing the antidandruff climbazole

In this study, the effect of the antidandruff climbazole on the rheology of hair shampoo containing Dead Sea (DS) salt was investigated. The presence of either DS salt or the climbazole led to increase in the shampoo viscosity. An optimum concentration was found where the viscosity of shampoo was maximum. In the absence of DS salt, the viscosity of hair shampoo increased with increasing the climbazole concentration to reach a maximum value at 1.0 wt%. Further addition of climbazole decreased the viscosity of shampoo. Adjusting the pH of the shampoo at 5.5 and 5.0 shifted the optimum climbazole concentration (corresponds to maximum viscosity) to 0.8 wt% and led to increase in the viscosity of shampoo. On the other hand, the addition of climbazole to the shampoo containing DS salt resulted in a decrease in shampoo viscosity. This decrease of shampoo viscosity became more pronounced with increasing the climbazole and/or DS salt concentrations. By controlling the pH of shampoo, an optimum formula of shampoo comprising both climbazole and DS salt and having maximum viscosity was obtained.     

检测赭曲霉毒素A(OTA)的酶联免疫吸附法(ELISA)体系的建立

本文分别以自制抗-OTA兔血清抗体(IgG)及鸡卵黄抗体(IgY)就影响检测OTA的ELISA(酶联免疫吸附法)体系因素进行了探讨,建立了检测OTA的ELISA方法。鸡卵黄抗体的最佳ELISA操作参数为:包被抗原浓度为7.5μg/ml,封阻剂浓度为2.5%,抗体稀释倍数为1000,酶标二抗稀释倍数为14000;兔血清抗体的最佳ELISA操作参数为:包被抗原浓度为2.5μg/ml,封阻剂浓度为2.5%,抗体稀释倍数为8000,酶标二抗稀释倍数为10000。IgG和IgY的检测灵敏度分别为10ng/ml和1ng/ml。     

Adverse effects of cosmetics and toiletries: a retrospective study in the general population

Of 1609 individuals who were interviewed and asked whether they had experienced adverse reactions to cosmetics or toiletry products in the preceding 5 years, 196 (12.2%) claimed to have experienced some such reaction.
Females ( n = 124) mostly attributed their complaints to soap (41%), facial creams (33%), deodorant (25%), shampoo (16%) and eye shadow (11%). Men ( n = 72) complained about adverse effects from soap (49%), aftershave (22%), deodorant (19%) and shower foam (12%). Both in women and in men, most reactions were localized on the face (60% resp. 33%), followed by the hands (19% resp. 21%) and the axillae (18% resp. 14%). The majority of patients could solve the problem by stopping the use of the suspected product and purchasing a different brand. Nevertheless, medical consultation was sought by more than 30% of all patients.
Presumably, the majority of all adverse effects were caused by irritation; contact allergic reactions are infrequent.     

Phosphate diesters for dermatological use

A new formulation for dermatological use and hair care was studied. Phosphate diesters of the nonyl-phenol type in association with cocoimidazoline derivatives are compared with four commercial formulations. Foaming capacity, eye and skin irritation, keratin solubilization and defatting properties were studied. The mixture was the best foamforming formulation and the best defatting agent. Also, it was non-irritant for skin and eyes at adequate dilutions and did not dissolve keratin. It is a good formulation for dermatological use and hair care, due to its detergent properties. It ranks high among safe products.
Di-ésters phosphates à usage dermatologique     

In vitro cultured human skin cells as alternatives to animals for skin irritancy screening

In the last few years a lot of attention has been paid to the development of the in vitro models which would substitute for animals in cutaneous irritancy studies. These models explore either organ or explant cultures using freshly excised skin or serial cultures of isolated skin cells (epidermal keratinocytes or dermal fibroblasts). The organ or explant models are suitable only for short exposures of skin samples to the compounds tested and the use of it will always be restricted by the limited availability of fresh human skin. The model that uses submerged cultures of keratinocytes or fibroblasts permits the production of a large number of cells, and permits large scale toxicity screening tests with many substances, that can be applied in a broad concentration range. Since the stratum corneum is absent in conventional (submerged) keratinocyte culture systems, this model is mainly suited for testing of water soluble compounds and it is less suitable for poorly soluble compounds and for topical products consisting of complex formulations which are made of active ingredients and their vehicles. This shortcoming can be overcome by using ‘organotypic cultures’in which keratinocytes are grown at the air-liquid interface on a suitable dermal substrate. Under these conditions, the culture forms a multilayered epidermis showing an overall structure which resembles that of a native epidermis. The presence of a coherent stratum corneum layer in these cultures permits the application of potential irritants at concentrations and in formulations as applied in vivo. For the evaluation of toxicity a number of tests have already been developed: assessment of cell viability, changes in cell morphology, modulation of cell proliferation and differentiation, monitoring of membrane damage, the measurements of the uptake or incorporation of radioactive precursors, establishment of the modulation of cell metabolism, determination of the release of inflammatory mediators, etc. All these in vitro techniques are still in a state of validation as far as their predictive value for in vivo skin irritancy is concerned.     

Study of the fate of Bronopol and the effects of antioxidants on N-nitrosamine formation in shampoos and skin creams

The role of antioxidants in preventing nitrosamine ( N -nitroso diethanolamine) formation in shampoos and skin creams and the levels of Bronopol (2-bromo, 2-nitropropane-1, 3-diol) addition in relation to the resultant levels of N -nitroso compounds have been studied.
The addition of 0.05% BHT, α-tocopherol or ascorbate was effective in preventing nitrosamine formation (limit of determination: 2.5 μ g kg^-1) in products preserved with 0.01% Bronopol. At 0.05% Bronopol, 0.2% of all three antioxidants was effective in preventing nitrosamine formation but at 0.1% Bronopol only α-tocopherol was effective.
Levels of Bronopol found in shampoo formulations stored for 2 weeks at 37°C with initial concentrations of 0.005-0.1% were reduced by 64-86%, independent of the initial concentration. The levels in skin creams stored for extended periods at ambient temperature with 0.01% Bronopol initially were reduced by, on average, 60%.
High nitrite levels (NO₂^-) were found in formulations containing 0.01% Bronopol but no antioxidants (200-250 mg kg^-1). Detected levels of nitrite in skin cream formulations were greater than could be formed by stoichiometric decomposition of Bronopol. In formulations with both Bronopol and antioxidants the levels of nitrite were considerably reduced. When α-tocopherol rather than BHT or ascorbate was the antioxidant then the nitrite levels were appreciably higher but formation of N -nitroso diethanolamine was no greater.
High nitrite levels correlated with the formation of detectable levels of N -nitroso compounds in the absence of antioxidants.
Le rôle préventif du Bronopol et des antioxygènes dans la formation de la nitrosamine     

生防菌株洋葱伯克氏菌对草莓果实采后病害控制及安全性评价

采用自然腐烂法和伤口接种法研究生防菌株洋葱伯克氏菌Burkholderia contaminans(简称B-1),在不同发酵时间、处理方式和处理液对草莓果实采后贮藏的影响,并进行该生防菌的大鼠急性经口毒性实验、家兔皮肤及眼刺激性实验和豚鼠皮肤变态反应(致敏)实验。结果表明:与CK相比,生防菌B-1发酵培养24 h、采后果实浸泡3 min及稀释5倍的发酵液处理草莓果实,其采后腐烂率分别下降了55.04%、46.74%和39.98%,而且发酵原液、稀释5倍发酵液及菌悬液对草莓果实伤口接种灰霉病抑制率达100%。该生防菌的大鼠急性LD₅₀>5000 mg/kg,皮肤刺激强度和眼刺激积分指数(I.A.O.I)为0,眼刺激平均指数(M.I.O.I)48 h为0,皮肤变态反应(致敏)实验致敏率为0。本研究表明该细菌作为生防菌具有安全性,为该生防菌的生产性应用奠定基础。     

Compatibility testing in vitro: a comparison with in vivo patch test data

For safety reasons cosmetic formulations have to be assessed for putative side effects like skin irritation. The human patch test is an appropriate method for this type of assessment, but it cannot be used for the higher throughput needed for screening innovative formulations and a distinct assessment is often difficult. The aim of this study was therefore to investigate the feasibility of using an in vitro approach for compatibility testing to determine the irritancy of surfactants. Test samples were provided by the German Society of Cosmetic Chemists (DGK), which conducted a human patch test study with the same set of samples in parallel. This gave us the unique opportunity to correlate in vitro with in vivo data. To assess irritant effects in vitro , reconstructed human epidermis was exposed to seven coded test samples consisting of individual anionic surfactants, blends of surfactants, and controls. A multiple endpoint analysis was established comprising the viability, cytotoxicity, histology, cytokine release and differential gene expression. Using this test strategy, a very good correlation was determined for our in vitro assessment of compatibility with a theoretical ranking and the human patch test data.     

A review of shampoo surfactant technology: consumer benefits,raw materials and recent developments

Surfactants form the core of all shampoo formulations, and contribute to a wide range of different benefits, including cleansing, foaming, rheology control, skin mildness and the deposition of benefit agents to the hair and scalp. The purpose of this review was to assist the design of effective, modern, shampoo surfactant technologies. The mechanisms through which surfactants help deliver their effects are presented, along with the appraisal techniques through which surfactant options can be tested and screened for product development. The steps that should be taken to select the most appropriate blend of surfactants are described, and useful information on the most widely used surfactants is provided. The review concludes with an examination of recent developments in ‘greener’ surfactants, ‘sulphate‐free’ technologies and structured liquid phases for novel sensory properties and for suspending benefit agents.