Safety Assessment of Ginkgo biloba-Derived Ingredients as Used in Cosmetics.

The Expert Panel for Cosmetic Ingredient Safety (Panel) assessed the safety of 10 Ginkgo biloba-derived ingredients, which are most frequently reported to function in cosmetics as skin conditioning agents or antioxidants. The Panel reviewed the available data to determine the safety of these ingredients. Because final product formulations may contain multiple botanicals, each containing the same constituents of concern, formulators are advised to be aware of these constituents and to avoid reaching levels that may be hazardous to consumers. The Panel was concerned about the presence of ginkgolic acid in cosmetics. Industry should use good manufacturing practices to limit impurities. The Panel concluded that 5 Ginkgo biloba leaf-derived ingredients are safe in the present practices of use and concentration described in this safety assessment when formulated to be non-sensitizing; data are insufficient to determine the safety of the remaining 5 ingredients under the intended conditions of use in cosmetic formulations.

Safety Assessment of Hops as Used in Cosmetics.

The Expert Panel for Cosmetic Ingredient Safety (Panel) assessed the safety of Humulus Lupulus (Hops) Extract (reported functions include antimicrobial agent and hair conditioning agent) and Humulus Lupulus (Hops) Oil (reported function is fragrance). The Panel reviewed the relevant data related to these ingredients. Because final product formulations may contain multiple botanicals, each containing the same constituents of concern, formulators are advised to be aware of these constituents and to avoid reaching levels that may be hazardous to consumers. For these ingredients, the Panel was concerned about the presence of 8-prenylnaringenin, ß-myrcene, and quercetin in cosmetics, which could result in estrogenic effects, dermal irritation, and genotoxicity, respectively. Industry should use current good manufacturing practices to limit impurities and constituents of concern. The Panel concluded that Humulus Lupulus (Hops) Extract and Humulus Lupulus (Hops) Oil are safe in cosmetics in the present practices of use and concentration when formulated to be non-sensitizing.

A botanical reference set illustrating a weight of evidence approach for skin sensitization risk assessment.

Recent years have seen an increase in the use of botanicals and natural substances (BNS) in consumer products such as cosmetics and household care products. Most work conducted to date to assess botanicals for human safety has focused their use as dietary supplements and thus on systemic toxicity. However, the induction of skin sensitization is a possible adverse effect of natural products in particular those that come into skin contact, especially for cosmetics that remain on the skin and are not rinsed off following use. Assessments of BNS ingredients are often challenging for a number of reasons: the BNS are complex mixtures that can be of mostly unknown composition; the composition can be highly variable even within the same plant species and dependent on how processed; the physical form of the BNS raw material can vary from a highly concentrated powdered extract to a liquid extract containing only a small percentage of the BNS; testing of the BNS raw materials in New Approach Methods (NAM) has uncertainty as these methods are often not developed or validated for complex mixtures. In this study, a reference set of 14 selected BNS which span the range of skin sensitization potential was complied. These data were used in a Weight of Evidence (WoE) approach to evaluate their skin sensitization potential with each of the data rich BNS being classified as either having strong evidence of inducing skin sensitization based on human topical use history, animal data, clinical data, composition data and NAM data, or having some but more limited (weak) evidence of inducing skin sensitization, or having strong evidence of no skin sensitization potential. When available data have sufficient potency related information, sensitization potency assessment is also provided based on WoE, classifying these BNS as either strong, moderate, or weak sensitizers, or non-sensitizers. An outline for a BNS skin sensitization risk assessment framework is proposed starting with exposure-based waiving and WoE assessment for higher exposures. In addition to demonstrating the application of the WoE approach, the reference set presented here provides a set of 'data rich' botanicals which cover a range of sensitization potencies that could be used for evaluating existing test methods or aid in the development of new predictive models for skin sensitization.

Amended Safety Assessment of Mentha piperita (Peppermint)-Derived Ingredients as Used in Cosmetics.

The Expert Panel for Cosmetic Ingredient Safety (Panel) reviewed the safety of M piperita (peppermint)-derived ingredients. The Panel reviewed data relevant to the safety of these ingredients. Because final product formulations may contain multiple botanicals, each containing the same constituent(s) of concern, formulators are advised to be aware of these constituents and avoid reaching levels that may be hazardous to consumers. Industry should continue to use good manufacturing practices to limit impurities that could be present in botanical ingredients. The Panel concluded that M piperita (Peppermint) Oil, Extract, Leaf, and leaf-derived ingredients are safe in cosmetics in the present practices of use and concentration when formulated to be non-sensitizing, and that the available data are insufficient for determining that M piperita (Peppermint) Flower/Leaf/Stem Extract, M piperita (Peppermint) Flower/Leaf/Stem Water, and M piperita (Peppermint) Meristem Cell Culture are safe under the intended conditions of use in cosmetic formulations.

The Safety Assessment of Cosmetic Perfumes by Using In Chemico and In Vitro Methods in Combination with GC-MS/MS Analysis.

Animal testing has been prohibited for the safety assessment of cosmetic ingredients or finished products. Thus, alternative non-animal methods, followed by confirmatory clinical studies on human volunteers, should be used as the sole legally acceptable approach within the EU. The safety assessment of cosmetic products requires the involvement of multiple scientific disciplines, including analytical chemistry and biomedicine, as well as in chemico, in vitro and in silico toxicology. Recent data suggest that fragrance components may exert multiple adverse biological effects, e.g. cytotoxicity, skin sensitisation, (photo)genotoxicity, mutagenicity, reprotoxicity and endocrine disruption. Therefore, a pilot study was conducted with selected samples of fragrance-based products, such as deodorant, eau de toilette and eau de parfum, with the aim of integrating results from a number of alternative non-animal methods suitable for the detection of the following toxicological endpoints: cytotoxicity (with 3T3 Balb/c fibroblasts); skin sensitisation potential (in chemico method, DPRA); skin sensitisation potential (LuSens in vitro method, based on human keratinocytes); genotoxicity potential (in vitro Comet assay with 3T3 Balb/c cells); and endocrine disruption (in vitro YES/YAS assay). The presence of twenty-four specific known allergens in the products was determined by using GC-MS/MS. The strategies for estimation of the NOAEL of a mixture of allergens, which were proposed by the Scientific Committee on Consumer Products in their 'Opinion on Tea tree oil' document and by the Norwegian Food Safety Authority in their 'Risk Profile of Tea tree oil' report, were used as models for the NOAEL estimation of the mixtures of allergens that were identified in the individual samples tested in this study.

Safety Assessment of Apple-Derived Ingredients as Used in Cosmetics.

The Expert Panel for Cosmetic Ingredient Safety (Panel) reviewed the safety of 26 apple-derived ingredients, which reportedly function mostly as skin conditioning agents in cosmetic products. Because apple-derived ingredients may be obtained from different apple cultivars, the composition of ingredients derived from different cultivars should be similar to that of ingredients reviewed in this safety assessment. Additionally, industry should continue to use good manufacturing practices to limit impurities that could be present in botanical ingredients. The Panel reviewed the available data to determine the safety of these ingredients and concluded that 21 of these ingredients are safe in cosmetics in the present practices of use and concentrations described in this safety assessment. However, the Panel also determined that the available data are insufficient to determine the safety of Pyrus Malus (Apple) Root Extract, Pyrus Malus (or Malus Domestica) (Apple) Stem Extract, Malus Domestica (Apple) Callus Extract, and Malus Domestica (Apple) Oil.

Safety Assessment of Centella asiatica-Derived Ingredients as Used in Cosmetics.

The Expert Panel for Cosmetic Ingredient Safety (Panel) assessed the safety of 9 Centella asiatica-derived ingredients, which reportedly function primarily as skin conditioning agents in cosmetic products. The Panel reviewed relevant data relating to the safety of these ingredients. The Panel concluded that Centella Asiatica Extract, Centella Asiatica Callus Culture, Centella Asiatica Flower/Leaf/Stem Extract, Centella Asiatica Leaf Cell Culture Extract, Centella Asiatica Leaf Extract, Centella Asiatica Leaf Water, Centella Asiatica Meristem Cell Culture, Centella Asiatica Meristem Cell Culture Extract, and Centella Asiatica Root Extract are safe in the present practices of use and concentration in cosmetics, as described in this safety assessment, when formulated to be non-sensitizing.

Safety Assessment of Plant-Derived Proteins and Peptides as Used in Cosmetics.

The Expert Panel for Cosmetic Ingredient Safety (Panel) reviewed the safety of 19 plant-derived proteins and peptides, which function mainly as skin and/or hair conditioning agents in personal care products. The Panel concluded that 18 plant-derived proteins and peptides are safe as used in the present practices of use and concentration as described in this safety assessment, while the data on Hydrolyzed Maple Sycamore Protein are insufficient to determine safety.

Causes and countermeasure for blank absorbance increase in the ROS assay.

A reactive oxygen species (ROS) assay is an in chemico photoreactivity test listed in ICH S10 guideline and OECD Test Guideline No. 495. We currently utilize the ROS assay to assess the photosafety of cosmetic ingredients. We have recently confronted a problem that there was an absorbance increase of blank assessing superoxide anion generation after irradiation, whereas this did not occur in the negative control (sulisobenzone), leading to a dissatisfaction of the acceptance criteria. Therefore, we aimed to investigate the causes and find countermeasures. No significant effects of impurities and manufacturer differences of sodium phosphate and DMSO on blank absorbance increases were observed. In contrast, when Cu2+ was added to the buffer, the increase of blank absorbance after irradiation did not occur. We then confirmed the dose-response relationship and found that adding 0.1 µM of Cu2+ (corresponding to 6 ppb of Cu2+) was sufficient in suppressing the blank absorbance increase, suggesting the need of Cu2+ supplementation to the buffer. Finally, we confirmed that the ROS assay using the buffer supplemented with 0.1 µM of Cu2+ obtained stable test results by using 17 proficiency chemicals listed in TG 495. Our results suggest that the modified ROS assay protocol would be useful for obtaining stable test results.

Epidemiology, clinical presentation, treatment, and follow-up of chronic mercury poisoning in China: a retrospective analysis.

BACKGROUND: There are no reports on the incidence of chronic mercury poisoning in a large population in China. This study investigated the epidemiology, clinical manifestations, treatment, and follow-up of Chinese patients with chronic mercury poisoning. METHODS: Data for 288 mercury poisoning patients were collected at our hospital from July 2014 to September 2019, including sex, age, admission time, blood mercury content, urine mercury content, creatinine, urinary mercury/creatinine ratio, 24-h urinary protein levels, electromyography (EMG) findings, renal biopsy, and follow-up. Patient characteristics were evaluated by statistical and correlation analyses. RESULTS: First, mercury poisoning in China mainly occurred through occupational exposure and the inappropriate use of mercury-containing cosmetics and Chinese folk remedies (CFRs). Second, the most common symptoms were nervous system (50.3 %), kidney (16.4 %) and breathing (8.0 %). Mercury poisoning-induced Nephrotic syndrome (NS) and peripheral neuropathy are common long-term complications. The complications of occupational and cosmetics-induced mercury poisoning are consistent with international belief. However, the NS caused by CFRs is mainly membranous nephropathy and the probability of peripheral neuropathy caused by CFRs is higher than other pathogens. Third, follow-up data shows that 13 patients with EMG-confirmed neurological injury, 10 showed full recovery after 38.50 ± 8.03 months. Furthermore, among 18 patients with NS, 15 had normal urine protein and serum albumin levels after 22.67 ± 10.26 months. CONCLUSIONS: Regulation of skin-lightening cosmetic products, safety surveillance of CFRs, and prevention and control of occupational exposure must be improved to decrease the incidence of mercury poisoning in China.

Saponin fraction from Sapindus mukorossi Gaertn as a novel cosmetic additive: Extraction, biological evaluation, analysis of anti-acne mechanism and toxicity prediction.

ETHNOPHARMACOLOGICAL RELEVANCE: Sapindus mukorossi Gaertn. (S. mukorossi), known as 'mu huan zi' in Chinese folklore, belongs to the family Sapindaceae and it has been traditionally used for treating coughing and excessive salivation, removing freckle, whitening skin, etc. Evidence-based medicine also verified the antimicrobial, anti-tyrosinase and anti-acne activity of S. mukorossi extract, suggesting that it has the potential to be a pharmaceutical and cosmetic additive. AIM OF THE STUDY: The present study was intended to evaluate the freckle-removing and skin-whitening activities of S. mukorossi extracts, and further analyzing the potential anti-acne mechanism. METHODS: Saponin fractions were purified by using the semi-preparative high-performance liquid chromatography, and their antibacterial activity was detected against Propionibacterium acnes (P. acnes), which was the leading cause of inflamed lesions in acne vulgaris. The anti-lipase and anti-tyrosinase activities were assayed using a commercial kit, while the potential anti-acne mechanism was predicted on the basis of the network pharmacology. Active components of saponin fraction were identified by HPLC-MS analysis. Furthermore, the different toxicity level of compounds was predicted according to the quantitative structure-activity relationship, and the first application of crude extract and saponin fraction to facial masks was analyzed based on the comprehensive evaluation method. RESULTS: The saponin fraction (F4) purified from the fermentation liquid-based water extract (SWF) showed the best antibacterial activity against P. acnes ATCC 6919 with the MIC of 0.06 mg/mL, which was 33-fold of its parent SWF (with the MIC of 2.0 mg/mL). Compared with SWF, the application of F4 caused greater inhibition rates on lipase and tyrosinase. Chemical constituents of F4 were evaluated, from which four oleanane-type triterpenoid saponins were detected to contribute to the above biological activities of F4. The mechanism of the four compounds on anti-acne was predicted, and seven targets such as PTGS2 and F2RL1 were obtained to be important for the treatment of acne. The four compounds were also predicted to have different levels of toxicity to various species, and they were not harmful to rats. Besides, F4 and SWF were applied to facial masks and there was no significant influence on the physicochemical properties including pH, stability, and sensory characteristics. CONCLUSION: This work demonstrated that oleanane-type triterpenoid saponins were speculated to contribute to the skin-whitening, freckle-removing, and anti-acne activities of F4. These findings will facilitate the development of the S. mukorossi extract and the allied products as the new and natural anti-acne agent and cosmetic additives.

Evaluation of the safety of hair essence containing 0.05% purified bee venom on the skin and eyes of rabbits.

OBJECTIVE: To investigate the safety of hair essence containing 0.05% purified bee venom (HE-PBV) on the skin and eyes of New Zealand White rabbits. METHODS: HE-PBV which contained 0.05% PBV, purified water, and glycerin, was used as the test substance. The skin-irritation test (SIT) and eye-irritation test (EIT) were conducted according to the Draize method. On the SIT, HE-PBV (0.5 mL) dropped gauze was attached both intact and abraded skin for 24 h. The other side of the skin was used as control. After 24 and 72 h, the treatment site was observed and scored according to evaluation criteria for skin reactions. On the EIT, the rabbits were divided into two groups: eye-washed (three rabbits) and non-eye-washed (six rabbits). HE-PBV (0.1 mL) was squirted into the right eye of rabbits. The left eye was untreated and used as a control. Then, 20-30-s later, the eyes of rabbits in the eye-washed group were washed with ~50 mL of physiologic (0.9%) salt solution. Then, 1, 2, 3, 4 and 7 d after the start of the EIT, the eyes and behavior of the rabbits were observed. The degree of eye irritation elicited by HE-PBV was determined in three steps and then the criteria of the classification of eye-irritation scores. RESULTS: The SIT revealed erythema and edema at the site of HE-PBV application. At 72 h, the body weight of rabbits was reduced slightly, but other symptoms (except erythema and edema) were not observed. The Primary Irritation Index score was 0.6, and HE-PBV was deemed to be a slight irritant. The EIT did not show mortality or body-weight fluctuation, but hyperemic conjunctiva and eyelid closure were noted after HE-PBV administration. Except for these results, the score for the ophthalmic response on days 1, 2, 3, 4 and 7 was 0, and HE-PBV was deemed to be a non-irritant. CONCLUSION: These data suggest that HE-PBV did not elicit eye irritation, but was a slight irritant to the skin of rabbits; the latter slight would have been due to the excipients used in manufacture of the hair essence because PBV has been shown to be safe.

Cosmetics with hormonal composition for bioindicators Artemia salina L. and Allium cepa L. toxic potential.

The emerging pollutants cover a wide range of synthetic chemicals that are indispensable to modern society but with little known effects for aquatic animals and for people who consume polluted waters with these products. Gels manipulated with hormones are widely used for hormone replacement, muscle growth, among other purposes. However, only a small part of these hormones are absorbed into the skin, and so these can be transferred to the domestic sewage during the washing of exposed body regions. Thus, the aim of this study was to evaluate the toxicity levels for the bioindicator Artemia salina L., and cytotoxicity and mutagenicity for the bioindicator Allium cepa L. of gels handled with 1% testosterone and 0.1% estradiol. Data from immobile/dead A. salina numbers after 24 h of exposure showed that the highest concentrations of testosterone (10 and 25 µg/mL) and the highest concentration of estradiol (15 µg/mL) were toxic to this bioindicator. For the bioindicator A. cepa, mitotic indices and chromosomal aberrations did not indicate statistical differences between the groups treated with the testosterone gels (1, 10, 50 µg/mL) and estradiol (0.03, 0.30, 1.5 µg/mL) and the control group. However, all concentrations of the testosterone-containing gel decreased the percentage of cell division in relation to the time 0 h of each treatment and to the time 24 h of the negative control. Therefore, it is concluded that rivers or aquatic environments can be polluted if wastewater with the toxic concentrations found of these hormonal gels is discarded without previous treatment, compromising the life of organisms that live there. And, it encourages the development of techniques for treating sewage and water to reduce/eliminate the hormones present in them.

Safety Assessment of Citrus-Derived Peel Oils as Used in Cosmetics.

The Cosmetic Ingredient Review Expert Panel assessed the safety of 14 citrus-derived peel oil ingredients and concluded that these ingredients are safe for use in cosmetic products when finished products, excluding rinse-off products, do not contain more than 0.0015% (15 ppm) 5-methoxypsoralen, and when formulated to be nonsensitizing and nonirritating. The citrus-derived peel oil ingredients are most frequently reported to function in cosmetics as fragrances and/or skin conditioning agents. The Panel reviewed the available animal and clinical data to determine the safety of these ingredients. Because final product formulations may contain multiple botanicals, each containing the same constituents of concern, formulators are advised to be aware of these constituents and to avoid reaching levels that may be hazardous to consumers. Industry should use good manufacturing practices to limit impurities that could be present in botanical ingredients.

Threshold of Toxicological Concern (TTC) for Botanical Extracts (Botanical-TTC) derived from a meta-analysis of repeated-dose toxicity studies.

Threshold of Toxicological Concern (TTC) is a promising approach for evaluating the human health risk for systemic toxicity when there is a lack of toxicological information. The threshold for systemic toxicity is reportedly 1800, 540, and 90 µg/day for Cramer I-III chemical structures, according to Munro's structural decision tree, and 0.15 µg/day for genotoxic compounds. However, the concept of TTC has been developed for single substances; therefore, the applicability of TTC for mixtures remains unclear. To expand application of probability approach for mixtures, a validation study using the point of departures (PoDs) derived from mixtures is required. In the present study, we investigated novel TTC of botanical extracts (Botanical-TTC) for cosmetics from a meta-analysis based on the PoDs derived from repeated dose toxicity testing in botanical extracts. Accordingly, 213 PoDs were determined by repeated-dose toxicity studies and divided using a default uncertainty factor of 100 combined with the extrapolation factor of study duration to calculate the derived-no-effect-level (DNEL) and derived-minimal-effect-level (DMEL). The minimum DNEL/DMEL was 1.6-fold higher than the Cramer III TTC. In addition, because human health risk below the 1 st percentile value (663 µg/day) was considered as extremely limited, the exposure level can be proposed as Botanical-TTC.

Current Methods for the Discovery of New Active Ingredients from Natural Products for Cosmeceutical Applications.

Cosmeceuticals are designed to serve a dual purpose: to provide desired esthetical effects and to treat dermatological conditions. Natural products derived from plants and marine organisms are a novel source of potential cosmeceutical active ingredients for incorporation into new formulations due to consumer demands. Contrary to common perceptions, most regulatory agencies do not view cosmeceuticals as being a separate category from cosmetics; thus, these products are not regulated accordingly, thereby forcing the consumer to rely on the self-regulatory policies of the cosmetics industry. Cosmeceuticals are advertised as having capabilities that include anti-aging, anti-acne, solar-protective, wound healing, and skin whitening. Such traits normally comprise several biological activities. In order to ensure the safety and efficacy of these products, active ingredients employed in the formulations must undergo a series of tests. In this review, in vitro (enzymatic and cellular) and in vivo tests employed to evaluate the potential of new cosmeceutical active ingredients are discussed, and new trends that are being explored by the cosmeceutical industry are described.

Spirulina, Palmaria Palmata, Cichorium Intybus, and Medicago Sativa extracts in cosmetic formulations: an integrated approach of in vitro toxicity and in vivo acceptability studies.

Background/Aims: The selection of suitable raw materials in the cosmetic research and development is a key point, not only in order to obtain the expected results but also to avoid undesirable side effects. This study evaluated the in vitro toxicity potential of four different plant extracts and their in vivo acceptability studies. Methods: Spirulina, Palmaria palmata, Cichorium intybus and Medicago sativa extracts were analysed alone or in combination and added in cosmetic formulations. The in vitro toxicity evaluation, Hen's Egg Chorioallantoic Membrane Test (HET-CAM) and 3T3 NRU phototoxicity test were performed to evaluate in vitro potential ocular irritation and photo safety, respectively. Twenty subjects were enrolled in the acceptability studies, who were evaluated for the absence of harmful effects of the formulation by visual assessment and by transepidermal water loss, a biophysical technique, for 30 days. Results: HET-CAM assay showed that the studied extracts added to a gel-cream formulation had no irritant potential. In addition, the combination of Palmaria palmata, alfalfa and chicory extracts did not show phototoxic potential in vitro. Acceptability studies showed that the formulation containing the four extracts combined did not provoke any transepidermal water loss (TEWL) alteration, sensory irritation or erythema in the forearms for the period of analysis. Conclusion: The studied active ingredients, alone or in combination, present no cytotoxicity potential and when added to a gel-cream formulation had no irritant potential in vitro. These results predicting no harmful effects were confirmed in the acceptability tests, which showed no alteration on skin barrier function and no report of irritation perception of sign of erythema, suggesting the potential of these extracts for the development of safe cosmetic products.

Safety Assessment of Plant-Derived Fatty Acid Oils.

The Cosmetic Ingredient Review Expert Panel (Panel) assessed the safety of 244 plant-derived fatty acid oils as used in cosmetics. Oils are used in a wide variety of cosmetic products for their skin conditioning, occlusive, emollient, and moisturizing properties. Since many of these oils are edible, and their systemic toxicity potential is low, the review focused on potential dermal effects. The Panel concluded that the 244 plant-derived fatty acid oils are safe as used in cosmetics.

Use of the Bovine Udder Skin model to evaluate the tolerability of Mesem cosmetic cream.

Observational studies of Mesem cream (based on Mesembryanthemum crystallinum L. plant extract) found that it had positive effects on skin hydration and smoothing of the skin. However, some patients reported skin irritation effects. The current study evaluated the skin tolerability of Mesem cream, as compared to the carrier cream (without the active ingredient), by using the isolated perfused bovine udder skin model. The primary outcomes investigated were cytotoxicity (i.e. cell viability), assessed with the MTT assay, and irritancy and inflammation, assessed by measuring PGE2 tissue levels. A total reaction score was calculated by combining the results for each parameter. In the case of a single topical application, significant differences were found between the carrier cream and the Mesem cream. While the application of carrier cream resulted in low cytotoxicity (-8.4% change in viability, as compared to the untreated control), the Mesem cream was more cytotoxic (-18.7% change). In addition, one hour after application, PGE2 levels were higher in Mesem cream-treated skin, as compared to carrier cream-treated skin (16.6% versus 11.3%). Further experiments (tape-stripped skin and repeated application) also found significant differences between the two creams in the results obtained. Evaluation of the effectiveness, safety and tolerability of phyto-cosmetic products is important. Our results confirmed the findings of two previous human observational studies (the human patch test and open application study). Future experiments to understand the underlying principles of its effectiveness, safety and tolerability should include extracts of M. crystallinum L. juice, as well as the Mesem cream itself.