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Draft Screening Assessment for the Challenge
Benzene, 1,2-dimethoxy-4-(2-propenyl)- (Methyl eugenol) Chemical Abstracts Service Registry Number 93-15-2 Environment Canada Health Canada March 2010 SynopsisThe Ministers of the Environment and of Health have conducted a screening assessment of Benzene, 1,2-dimethoxy-4-(2-propenyl)-, (commonly called Methyl eugenol), Chemical Abstracts Service Registry Number 93-15-2. Methyl eugenol was identified in the categorization of the Domestic Substances List as a high priority for action under the Ministerial Challenge. Methyl eugenol was identified as a high priority as it was considered to pose an intermediate potential for exposure of individuals in Canada and it had been classified by the US National Toxicology Program on the basis of carcinogenicity. This substance was not considered to be a high priority for assessment of potential risks to the environment as it did not meet the ecological categorization criteria for persistence, bioaccumulation potential and inherent toxicity to aquatic organisms. Therefore, the assessment focuses principally on information relevant to the evaluation of human health. Methyl eugenol is an organic substance and is naturally-occuring in the essential oils of several plant species. These oils are extracted for use principally as flavour ingredients in food and beverages and as fragrance ingredients and emollients in personal care products. It is a component of a citronella oil based personal insect repellent registered for use in Canada. Based on information reported pursuant to section 71 of the Canadian Environmental Protection Act, 1999 (CEPA 1999), it was not reported to be manufactured in Canada in 2006, and there was less than 100 kg of the substance imported into the country in the same calendar year. Methyl eugenol is considered ubiquitous in air and water at very low concentrations. The predominant source of exposure to the general population is expected to result from its naturally-occuring presence in food and beverages, with smaller contributions from the use of personal care products and citronella oil based personal insect repellents. Based principally on the weight of evidence–based assessments of international or other national agencies, a critical effect for the characterization of risk to human health for methyl eugenol is carcinogenicity. In the standard 2-year carcinogenicity studies with rats and mice, methyl eugenol induced multiple types of tumours in both males and females in a dose-related manner. Of note, the significantly increased incidences of liver tumours were observed at the lowest dose tested in both rats and mice in the chronic studies. Methyl eugenol was genotoxic in a range of in vivo and in vitro assays, although it was not mutagenic in bacterial cells. Methyl eugenol bound to liver DNA and formed DNA adducts in vivo and in vitro. In addition, methyl eugenol caused gene mutation in the liver of transgenic animals and induced mutation of β-catenin gene in mouse liver tumours. While the mode of induction of tumours has not been fully elucidated, based on genotoxicity of methyl eugenol, it cannot be precluded that methyl eugenol induces tumours via a mode of action involving direct interaction with genetic material. Methyl eugenol is also associated with non-cancer effects in experimental animals including cytologic alteration, necrosis, hyperplasia, atrophy, organ or body weight changes in rats and mice. The critical non-cancer effect was reduced body weight or body weight gain. With respect to noncancer effects, comparison of the critical effect level with upper-bounding estimates of exposure to the general population from presence of methyl eugenol in use of personal care products and citronella oil based personal insect repellents results in margins of exposure that are considered adequate. On the basis of the carcinogenic potential of methyl eugenol, for which there may be a probability of harm at any exposure level it is proposed that methyl eugenol is a substance that may be entering the environment in a quantity or concentration or under conditions that constitute or may constitute a danger in Canada to human life or health. Based on its physical and chemical properties and limited experimental data, methyl eugenol does not meet the persistence and bioaccumulation criteria as set out in the Persistence and Bioaccumulation Regulations. In addition, both experimental and modelled toxicity data suggest that the substance is only moderately hazardous to aquatic organisms. Given the low quantity in commerce in Canada, the environmental concentration is predicted to be well below the predicted no effect concentration. On the basis of ecological hazard of methyl eugenol, it is proposed that this substance is not entering the environment in a quantity or concentration or under conditions that have or may have an immediate or long-term harmful effect on the environment or its biological diversity or that constitute or may constitute a danger to the environment on which life depends. Where relevant, research and monitoring will support verification of assumptions used during the screening assessment and, where appropriate, the performance of potential control measures identified during the risk management phase. Based on the information available, it is proposed that benzene, methyl eugenol meets one or more of the criteria set out in section 64 of the Canadian Environmental Protection Act, 1999. IntroductionThe Canadian Environmental Protection Act, 1999 (CEPA 1999) (Canada 1999) requires the Minister of the Environment and the Minister of Health to conduct screening assessments of substances that were prioritized during the categorization of substances on the Domestic Substances List to determine whether these substances present or may present a risk to the environment or to human health. Based on the information obtained through the categorization process, the Ministers identified a number of substances as high priorities for action. These include substances that
The Ministers therefore published a notice of intent in the Canada Gazette, Part I, on December 9, 2006 (Canada 2006), which challenged industry and other interested stakeholders to submit, within specified timelines, specific information that may be used to inform risk assessment and to develop and benchmark best practices for the risk management and product stewardship of those substances identified as high priorities. The substance methyl eugenol was identified as a high priority for assessment of human health risk because it was considered to present an IPE and had been classified by other agencies on the basis of carcinogenicity. The Challenge for methyl eugenol was published in the Canada Gazette on March 14, 2009 (Canada 2009). A substance profile was released at the same time. The substance profile presented the technical information available prior to December 2005 that formed the basis for categorization of this substance. As a result of the Challenge, submissions of information pertaining to the substance were received. Although methyl eugenol was determined to be a high priority for assessment with respect to human health, it did not meet the criteria for persistence, bioaccumulation potential or inherent toxicity to aquatic organisms. Therefore, this assessment focuses principally on information relevant to the evaluation of risks to human health. Screening assessments focus on information critical to determining whether a substance meets the criteria for defining a chemical as toxic as set out in section 64 of CEPA 1999[1] . Screening assessments examine scientific information and develop conclusions by incorporating a weight of evidence approach and precaution. This draft screening assessment includes consideration of information on chemical properties, hazards, uses and exposure, including the additional information submitted under the Challenge. Data relevant to the screening assessment of this substance were identified in original literature, review and assessment documents and stakeholder research reports and from recent literature searches, up to November 2009 for ecological effects and September 2009 for human health effects and exposure. Key studies were critically evaluated; modelling results may have been used to reach conclusions. Evaluation of risk to human health involves consideration of data relevant to estimation of (non-occupational) exposure of the general population, as well as information on health hazards (based principally on the weight of evidence assessments of other agencies that were used for prioritization of the substance). Decisions for human health are based on the nature of the critical effect and/or margins between conservative effect levels and estimates of exposure, taking into account confidence in the completeness of the identified databases on both exposure and effects, within a screening context. The draft screening assessment does not represent an exhaustive or critical review of all available data. Rather, it presents a summary of the existing critical information upon which the proposed conclusion is based. This draft screening assessment was prepared by staff in the existing substances programs at Health Canada and Environment Canada and incorporates input from other programs within these departments. The ecological and human health portions of this assessment have undergone external written peer review/consultation. Comments on the technical portions relevant to human health were received from Dr. Bernard Gadagbui, Toxicology Excellence for Risk Assessment; Dr. Michael Jayjock, The LifeLine Group; and Dr. Chris Bevans, CJB Consulting. The critical information and considerations upon which the draft assessment is based are summarized below. Substance IdentityFor the purposes of this document, benzene, 1,2-dimethoxy-4-(2-propenyl)- will be referred to as Methyl eugenol, derived from Philippine Inventory of Chemicals and Chemical Substances (PICCS). Information on the identity of methyl eugenol is summarized in Table 1. Table 1. Substance identity for Methyl eugenol
Source: NCI 2006 Physical and Chemical PropertiesTable 2 contains experimental and modelled physical and chemical properties of methyl eugenol that are relevant to its environmental fate. The models based on quantitative structure-activity relationships (QSARs) were used to generate data for some of the physical and chemical properties of methyl eugenol. These models are mainly based on fragment addition methods, i.e. they rely on the structure of a chemical. Based on its physical and chemical properties (Table 2), methyl eugenol is characterized by moderate water solubility (500 mg/L), moderate vapour pressure (modelled 1.6 Pa), low to moderate log Kow and log Koc (modelled, 3 and 2.7 respectively), and low Henry’s Law Constant (0.567 Pa·m3/mol). Table 2. Physical and chemical properties for Methyl eugenol
SourcesMethyl eugenol is a naturally occurring substance found in the essential oils of several plant species. The oils are extracted from plants by steam distillation or with organic solvents, typically for use as flavour or fragrance ingredients. The amount of Methyl eugenol in an essential oil extracted from a given type of plant varies with the variety, plant maturity at harvest, harvesting method, storage conditions and extraction method (Smith et al 2002). Methyl eugenol is also produced synthetically in small quantities. Annual production in the United States in 1990 was estimated to be 11,400 kg (NTP 2005a); in a more recent report, annual production in the United States was given as 77 kg (FAO/WHO 2009). There are currently four manufacturers of Methyl eugenol in the United States and three manufacturers elsewhere, but none in Canada (2009 email from SRI Consulting to Risk Assessment Bureau, Health Canada ; unreferenced). In response to the section 71 notice pursuant to CEPA 1999, no company reported the manufacture, import or use of Methyl eugenol in 2006 above the reporting thresholds. There are no other data on industrial activity with respect to Methyl eugenol in Canada . An extensive listing of the Methyl eugenol content of essential oils from different botanical sources is given in Appendix 2, where1, which reproduces a table of data from Burfield (2004). The concentrations of Methyl eugenol typically found in essential oils used in consumer products in Canada have not been quantified to date. UsesIn Canada , flavouring ingredients such as Methyl eugenol or essential oils containing Methyl eugenol can be added to any food that does not have a standard of identity and composition in the Food and Drug Regulations and to those foods that have a standard of identity and composition that allows for the addition of flavours to the food. Plant materials such as leaves, stems, and seeds containing Methyl eugenol may also be added to foods that do not have a regulatory standard, and to those that have a standard where there is provision for the addition of spices or seasoning. Some examples of common culinary herbs and spices that contain Methyl eugenol are basil, tarragon, lemon grass, bay leaf, nutmeg, allspice, cloves and mace. Methyl eugenol is also reported to have been found in oranges, bananas and grapefruit juice (Johnson and Abdo 2005; Smith et al 2002). Commercially prepared foods in which Methyl eugenol may be found include ice cream; baked goods such as cookies, pies, pastries and buns; puddings and other gelatine-based desserts; condiments, soups and sauces, especially pesto; various meat products; candy and chewing gum; and beverages made with spices and herbs containing Methyl eugenol (Council of Europe 2001). Methyl eugenol was classified as GRAS (Generally Recognized as Safe) by the Flavour and Extract Manufacturers Association (FEMA) in 1965 and that classification remained unchanged following a re-evaluation of Methyl eugenol by FEMA in 2001 (Smith et al 2002). In the United States , Methyl eugenol is a permitted food additive, provided that it is used in the minimum quantity required to produce its intended effect, and otherwise in accordance with all the principles of good manufacturing practice (US FDA 2001). In the European Union, EC Regulation 1334/2008, which applies from January 2011, prohibits the addition of Methyl eugenol to foods and restricts the concentration of Methyl eugenol in compound foods that have been prepared with flavourings or food ingredients with flavouring properties; however, if the only food ingredients with flavouring properties that have been added are fresh, dried or frozen herbs and spices, the maximum limits do not apply for methyl eugenol. Thus, pesto made with basil is a permitted food preparation, regardless of Methyl eugenol content. The permitted maximum concentrations range from 1 mg/kg in non-alcoholic beverages up to 60 mg/kg in soups and sauces (European Commission 2008). Some essential oils including citronella (Cymbopogon spp.), basil (Ocimum spp.), bay (Laurus nobilis) and tea tree (Melaleuca spp.) that may contain a high percentage of Methyl eugenol are used in fragranced consumer products such as personal care products and household cleaners. The European Union has conducted a risk assessment of Methyl eugenol in cosmetic and non-food products. Based on the findings of this assessment, Methyl eugenol is permitted in cosmetics as a component of plant extracts only. The permitted concentrations are as follows: 0.01% in fine fragrances, 0.004% in eau de toilette, 0.002% in a fragrance cream, 0.0002% in other leave-on products and oral hygiene products, and 0.001% in rinse-off products. Methyl eugenol may not be added as a pure chemical to cosmetics (European Commission 2000a, b). These concentration limits on the Methyl eugenol content of essential oils in cosmetic products were adopted by Canada and are outlined in the Cosmetic Ingredients Hotlist (Health Canada 2007). Citronella oil is an ingredient of personal insect repellent formulations in lotions and sprays applied to the skin. Not all citronella oil contains a high percentage of Methyl eugenol. In 2004, under the Pest Control Products Act, Health Canada conducted a re-evaluation of the safety of citronella oil use in personal insect repellents. The PMRA recommended that unless new data to address uncertainties and endpoints of concern were received, that citronella-based insect repellents that are applied to the skin be phased out of commerce in Canada . This recommendation was based in part on the toxicity of Methyl eugenol, a component of citronella oil (Health Canada 2004). Following a review by a scientific advisory panel, Health Canada recommended adopting the Methyl eugenol concentration limits proposed by the European Commission (Health Canada , 2008). Methyl eugenol is added as a fragrance in 15 pest control products in Canada , with a concentration range of 0.00233% to 0.005 %. However, these products are not registered for use on food. (2009 email from PMRA to Risk Assessment Bureau, Health Canada ; unreferenced). In the United States , Methyl eugenol is approved for use in fields and orchards in insect traps and lure products for control of fruit flies (US EPA 2006). In addition to its use in personal insect repellents, citronella oil is used in outdoor candles and torches as an area insect repellent. The tobacco of flavoured bidis and clove cigarettes has been analysed for a number of alkenylbenzene compounds, among them Methyl eugenol. The concentration of Methyl eugenol found in the cigarettes in this study ranged from not detected to 61 μg/g in strawberry-flavoured tobacco (Stanfill et al. 2003). The source of Methyl eugenol in flavoured tobacco is presumed to be in the flavouring and not the cured tobacco. In May 2009, the Government of Canada introduced amendments to the Tobacco Act to prohibit the selling of cigarettes, little cigars and blunt wraps (leaf-wrapped tobacco) with flavours and additives that taste like candy (Health Canada 2009). Essential oils are sold to individuals who choose to make their own preparations. Essential oils are used in a number of specialized applications such as aromatherapy, as ingredients of massage oils and in alternative medicine practices, among others. Methyl eugenol is a component of several essential oils which may be used in these practices (see Appendix 2). Considering the toxicity of Methyleugenol, the Natural Health Products Directorate has added new Toxicity Restrictions to the Natural Health Products Ingredients Database as follows:
Releases to the EnvironmentThere are insufficient data on which to base an estimate of releases of methyl eugenol to the environment. There are no known industrial sources of methyl eugenol releases to the Canadian environment; however, it is expected that, as in the United States (Barr et al. 2000), this substance is ubiquitous in air and water at low part per trillion levels The sources of Methyl eugenol in the environment have not been determined. Environmental FateBased on its physical and chemical properties (Table 2), the Level III fugacity model has been used to predict the environmental partitioning for methyl euganol, combinating with consideration of the half-lives in air (estimated as 23 hours, AOPWIN 2000), water (measured as 8 days, MITI 1992), soil (8 days as equivalent to it in water) and sediments (32 days as four times as it in water). The results from the modelling suggest that methyl eugenol is expected to mainly or predominantly reside in the environmental compartment to which it is released (Table 3). Table 3. Results of the Level III fugacity modelling (EQC 2003)
Persistence and Bioaccumulation PotentialEnvironmental Persistence In the atmosphere, methyl eugenol is not expected to undergo photolysis due to the lack of absorption in the environmental UV spectrum (>298 nm) (Meylan and Howard 1993). If released to air, the substance will be degraded by reaction with photochemically-produced hydroxyl radicals. The half-life for this reaction in air is estimated to be 5 hours (HSDB 1983-2009). In water, methyl eugenol is not expected to undergo hydrolysis in the environment due to the lack of hydrolysable functional groups. The empirical data from a biodegradation study (MITI 1992) show that there is approximately 90% ultimate biodegradation over 28 days in ready-biodegradation tests for methyl eugenol (Table 4a). The test indicates that the half-life of the substance in water is likely to be much shorter than 182 days (approximately 8 days, assuming first order degradation kinetics) and that it is therefore not likely to persist in water. Shaver and Bull (1980) identified a dissipation half-life of 34 hours in water and 16 hours in soil. Although they did not determine a mechanism for the dissipation, they speculated that the losses were probably a result of evaporation. Table 4a. Empirical data for degradation of methyl eugenol
In addition to experimental data on the degradation of methyl eugenol, a QSAR-based weight-of-evidence approach (Environment Canada 2007) was applied using the degradation models (see Table 4b below). Given the ecological importance of the water compartment, the fact that most of the available models apply to water and the fact that methyl eugenol is expected to be released to this compartment; biodegradation in water was primarily examined. Table 4b summarizes the results of available QSAR models for degradation of methyl eugenol in various environmental media. Table 4b. Modelled data for degradation of methyl eugenol
2 Output is a probability score In air, a predicted atmospheric oxidation half-life of 0.14 day (see Table 4b) demonstrates that methyl eugenol is likely to be rapidly oxidized. It is also predicted to react quickly with ozone, with a half-life value of 0.96 days. Therefore, the substance is considered to be not persistent in air. The BIOWIN (2008) aerobic biodegradation models (BIOWIN submodels 3, 4, 5 and 6) suggest that methyl eugenol biodegrades rapidly. The BIOWIN submodels 5 and 6 probability results are both greater than 0.3, the cut-off suggested by Aronson et al. (2006) to identify substances as having a half-life <60 days (based on the MITI probability models). The other two ultimate degradation models, TOPKAT and CATABOL, both predict that the substance may biodegrade rapidly in water. There is sufficient confidence, based on the experimental data with support from aerobic models (in Table 4b), to predict that there is significant biodegradation with a half-life in water that is far below 90 days and therefore the chemical is not considered persistent in water based on the criterion in this compartment. To extrapolate a half-life in water to the half-lives in soil and sediment, Boethling’s factors t½ water:t½ soil:t½ sediment = 1:1:4 (Boethling et al. 1995) were applied. Using the half-life in water of approximately 8 days (estimated from the MITI ready biodegradation test result - assuming first order kinetics) and the extrapolation factors, the half-lives in soil and sediment are estimated to be about 8 and 32 days, respectively, which are both far below 365 days. It is thus proposed that methyl eugenol is not persistent in soil or sediment. Based on the empirical and modelled data, it is proposed that methyl eugenol does not meet the persistence criteria in air, water, soil or sediment (half-life in air ≥ 2 days, half-lives in soil and water ≥182 days and half-life in sediment ≥ 365 days), as set out in the Persistence and Bioaccumulation Regulations (Canada 2000). Potential for Bioaccumulation Since no experimental bioaccumulation factor (BAF) or bioconcentration factor (BCF) data for methyl eugenol were available, a predictive approach was applied using available BAF and BCF models to estimate the bioaccumulation potential for the substance, as shown in Table 5 below. The modified Gobas BAF middle trophic-level model for fish predicted a bioaccumulation factor (BAF) of 77.27 L/kg, indicating that methyl eugenol does not have the potential to appreciably bioaccumulate nor to biomagnify in the environment. The result of the Gobas BCF model calculation also suggests a low bioconcentration potential for this substance (see Table 5). Note that the metabolic biotransformation potential for this substance was calculated from modelled BCF data, and was then used in calculating the QSAR-based Gobas BAF and Gobas BCF values. There is little difference, however, between the BCF and BAF estimates when metabolic biotransformation rates are included. This is because the predicted rates of biotransformation (i.e., 1.97 x 10-5 /d) are relatively inconsequential compared to the other rates of chemical elimination most notably gill elimination (i.e., 2.45 /d). The results of other BCF model calculations (OASIS Forecast 2005, and CPOPs 2008) shown in Table 5 below, add to the weight-of-evidence supporting the low bioconcentration potential of this substance with respect to the bioaccumulation criteria of 5,000 L/kg. Table 5. Modelled data of bioaccumulation for methyleugenol
Based on the model predictions on BAF and BCF, it is proposed that Methyl eugenol does not meet the bioaccumulation criteria (BAF or BCF > 5000) as set out in the Persistence and Bioaccumulation Regulations (Canada 2000). Potential to Cause Ecological HarmEcotoxicity studies for methyleugenol are available using fish, daphnia, and alga. The results of EC50/LC50 from acute studies ranged from 6 to 22 mg/L, while the no-observed-effect-concentrations (NOECs) ranged the 2.1 to 4.6 mg/L. In a chronic study, the EC50 and NOEC are 13 and 1.1 mg/L respectively. Data from aquatic toxicity studies of methyleugenol are listed in Table 6a. Table 6a. Empirical data for aquatic toxicity of methyleugenol
1 LC50 – The concentration of a substance that is estimated to be lethal to 50% of the test organisms. 2 EC50- The concentration of a substance that is estimated to cause some toxic sublethal effect on 50% of the test organisms. 3 NOEC – The No Observed Effect Concentration is the highest concentration in a toxicity test not causing a statistically significant effect in comparison to the controls. * Critical toxicity value to derive the predicted no effect concentration. ** AUG – Area Under Growth curve Besides the empirical toxicity data for methyleugenol, the predictive QSAR model ECOSAR (2008) was also used to estimate both acute and chronic effects of the substance on aquatic organisms. Table 6b contains predicted ecotoxicity values that were used in the QSAR weight-of-evidence approach for estimating aquatic toxicity (Environment Canada 2007). Table 6b. Modelled data for aquatic toxicity of methyleugenol (ECOSAR 2008)
The range of empirical toxicity values and the predicted aquatic toxicity (obtained from the QSAR model considered) indicates that methyl eugenol is not likely to cause acute harm to aquatic organisms at low concentrations (< 1 mg/L). Applying an assessment factor of 100 to the lowest empirical acute effect value of 6 mg/L – to account for inter- and intra-species variations in sensitivity, and to extrapolate from a laboratory-based endpoint to a chronic effect value in the field - results in a conservative predicted no effect concentration (PNEC) of 0.06 mg/L (see Robust Study Summary in Appendix 1). Being used as an insect attractant in combination with insecticide, methyleugenol is expected to exist as a vapour when released in the ambient atmosphere. Given the short half-life due to reactions with photochemically produced hydroxyl radicals, the substance will degrade fast and is not likely to a source of significant exposure. Given the very low quantity of this substance imported in Canada , a conservative aquatic exposure scenario was developed to estimate the potential release into the aquatic environment from industrial operations and the resulting aquatic concentration, using the 100-kg reporting threshold to represent the total amount used at a single facility. The predicted environmental concentration (PEC) is estimated to be 0.0006 mg/L (Environment Canada 2008). The resulting risk quotient (PEC/PNEC) is 0.01 (Environment Canada 2009), reflecting that methyleugenol is lacking in extreme toxicity and exposure, and therefore unlikely to cause ecological harm to aquatic organisms in Canada . With the exception of a predicted LC50 for an earthworm (Table 6b), no ecological effects information were found for this compound in media other than water. Based on the model predictions of environmental fate, if released to soil, there is the potential for soil-dwelling organisms to be exposed to methyleugenol. Given that emissions to soil are expected to be very low based on the use quantity, as well as the short dissipation half-life (measured as 16 hours at 22 oC) in that compartment, the exposure to methyleugenol in soil is anticipated to be limited. Based on this, and on the predicted LC50 for earthworms and the relatively low aquatic toxicity of this substance, significant ecological effects on soil organisms are unlikely. Uncertainties in Evaluation of Risk to EnvironmentThere is uncertainty associated with the exposure assessment. Although methyleugenol has been identified in various natural substances, there is no quantitative study assessing environmental (non-dietary) exposure. Given the low use quantity, confidence is high in the conservative nature of the exposure estimates, by using the 100 kg reporting threshold to predict the environmental concentration. Potential to Cause Harm to Human HealthExposure AssessmentEnvironmental Media and Diet Methyl eugenol was detected in the raw and partially treated effluent of one unbleached kraft mill at concentrations of 0.001–0.002 mg/L (Keith 1976), but not in the final effluent (Keith 1976). Air in the vicinity of insect traps baited with methyl eugenol was analysed for the presence of the substance 0–5 days after bait stations were set out. Methyl eugenol was detected in samples on days 0 and 1, but not on day 5, at distances of 1 and 25 m from the traps (Turner et al. 1989). No other reports of methyl eugenol levels in environmental media have been located. There are insufficient data with which to prepare an estimate of exposure to methyl eugenol from environmental media (air, water and soil); however, the exposure from these media is expected to be low. Estimates of the dietary intake of methyl eugenol from foods and beverages were prepared by the UK delegation to the Council of Europe Committee of Experts in Flavouring Substances for its 47th session in October 2000. The overall average and 97.5th-percentile intakes were estimated to be 190 and 530 µg/kg body weight (kg-bw) per day, respectively, for consumers only. These estimates were calculated using dietary intake data from a British survey of daily food and beverage consumption. For each food or beverage category, the highest level of methyl eugenol in that category was used to estimate dietary intake of methyl eugenol. The delegation report stated that these numbers were likely to be overestimates (Council of Europe 2001). Smith et al. (2002) prepared estimates of daily dietary intake of methyl eugenol and estragole in one of a series of safety evaluations by the Expert Panel of FEMA. The researchers used data on the annual volumes of plant materials with methyl eugenol-containing essential oils, methyl eugenol-containing essential oils and neat methyl eugenol imported and consumed in the United States in 1999. These data were combined with the average methyl eugenol content of essential oil derived from each plant type. Dietary intake of methyl eugenol was calculated by considering the intake of methyl eugenol from traditional food (principally spices), from essential oils added as flavour ingredients and from neat methyl eugenol as a flavouring substance. The estimated mean dietary intake of methyl eugenol for consumers only was 8 µg/kg-bw per day. The Joint Food and Agriculture Organization of the United Nations (FAO)/World Health Organization (WHO) Expert Committee on Food Additives (JECFA) published an evaluation of a group of alkoxy-substituted allylbenzenes, including methyl eugenol, in which the FEMA population-based estimation methodology as well as FEMA trade data were used to derive an estimate of the maximum dietary intake of methyl eugenol in the United States of 424 µg/person per day or about 6–8 µg/kg-bw per day for an adult (FAO/WHO 2009). The concentration of methyl eugenol in basil is highly variable and the use of basil to make pesto may occasionally result in the ingestion of a large amount of methyl eugenol. In the estimate of daily dietary intake of methyl eugenol from basil, Smith et al (2002) used an average concentration of methyl eugenol in dried basil of 2.6% and in fresh basil of 0.11%. Miele et al (2001) reported that the concentration of Methyl eugenol in essential oil extracted from Ocimum basilicum cv Genovese Gigante ranged from 5.5% to 100% and estimated that the intake from a single serving of pasta with pesto could reach 250 μg/kg-bw per meal for adults and 500 μg/kg-bw per meal for children, based on a concentration of methyl eugenol in basil oil of about 40%. Consumer Products Methyl eugenol is not permitted to be intentionally added as an ingredient in personal care products and is present only as a naturally occurring component of essential oils. Essential oils which may contain methyl eugenol are used in the formulation of thousands of personal care products in Canada (CNS, 2009). The Cosmetic Ingredient Hotlist details the maximum concentration of methyl eugenol in essential oils used in personal care products to: 0.01% in fine fragrances, 0.004% in eau de toilette, 0.002% in a fragrance cream, 0.0002% in other leave-on products and oral hygiene products, and 0.001% in rinse-off products (Health Canada 2007). The potential exposure to methyl eugenol from the use of personal care products made with essential oils containing methyl eugenol was derived using consumer exposure modelling software (ConsExpo 2006). A representative calculation is provided in Appendix 3. Based on information from notifications to Health Canada (CNS, 2009) upper-bounding estimates of systemic exposure to methyl eugenol from use of certain personal care products were derived and are shown for females in Table 7. Table 7. Estimated adult female systemic exposure of methyl eugenol from personal care products containing methyl eugenol. based on ConsExpo (ConsExpo 2006)
For adult women, the estimated daily systemic exposure to methyl eugenol resulting from dermal exposure only from the aggregate use of four types of personal care products (body lotion, face moisturizer, skin cleanser and fragrance) formulated with various essential oils containing methyl eugenol is 1.5 µg/kg-bw per day. This estimate assumes a dermal absorption of methyl eugenol of 40% for products left on the skin (European Commission 2000b) and a permeability coefficient of 0.0221 cm/hour for skin cleanser that is washed off (DERMWIN 2000). While the estimated exposure of adult women was the only scenario presented in the current screening assessment, the estimates of exposure from the use of personal care products are not expected to differ appreciably across age groups. As previously noted, the concentration of methyl eugenol in plant-derived material is quite variable, and there is significant uncertainty associated with these estimates, precluding the need to characterize exposures to other sub-populations. In an assessment of human exposure to methyl eugenol prepared by the European Cosmetics Association (COLIPA), for the European Commission, a lower estimate of the exposure to methyl eugenol from fragranced cosmetics was presented, in part based on a concentration of methyl eugenol of only 0.05% in the essential oils. Neither estimate includes exposure arising from the use of dental or oral hygiene products. Clove flower oil is licensed for sale in Canada as a non-prescription dental analgesic (LNHPD 2009). Methyl eugenol was detected in 98% of 206 adult human serum samples analysed in the Third National Health and Nutrition Examination Survey, an indication that human exposure in the United States is ubiquitous. The 5-95% distribution was 5-78 ng/kg in serum. It can be concluded that methyl eugenol is broadly present in human serum, however, the concentrations are extremely low. During the laboratory analysis phase of this work, it was determined that methyl eugenol was present in laboratory air and both distilled and bottled water at low parts per trillion levels. The researchers concluded that methyl eugenol is ubiquitous in air and water, albeit at very low levels and that human exposure arises from multiple sources (Barr et al. 2000). Confidence is high that the predominant sources of exposure to methyl eugenol for Canadians are diet and personal care products. While the highest concentration of methyl eugenol suggested for use in cosmetics is outlined in the Cosmetic Ingredient Hotlist, there are no data to characterize actual levels of methyl eugenol typically found in essential oils used in personal care products. There are no Canadian data on dietary consumption of methyl eugenol. Exposure to methyl eugenol via cleaning products is likely a minimal contributor to overall exposure. Health Effects AssessmentAppendix 4 contains a summary of the available health effect information for Methyl eugenol. The US National Toxicology Program (NTP) classified Methyl eugenol as reasonably anticipated to be a human carcinogen based on sufficient evidence of carcinogenicity in experimental animals (NTP 2000a,b; NTP 2005a). The International Agency for Research on Cancer (IARC), has not assessed methyl eugenol for carcinogenicity. However, the structurally related allylbenzene, safrole, was classified by the IARC as possibly carcinogenic to humans (Group 2B) and was listed as reasonably anticipated to be a human carcinogen by the US NTP (IARC 1976; NTP 2005b). In animal studies, methyl eugenol induced tumours in two species, in both genders, and at multiple sites. In the standard 2-year NTP carcinogenicity studies with rats and mice, methyl eugenol induced multiple types of tumours in liver and neuroendocrine tumours in the glandular stomach in both males and females in a dose-related manner. In the NTP studies, significantly dose-melated increased incidences of hepatocellular carcinoma or adenoma were observed in male Fischer 344 rats (7/50, 14/50, 28/50, 43/50, 45/50 for 0, 37, 75, 150 or 300 mg/kg-bw per day, respectively); and in female Fischer 344 rats (1/50, 8/50, 14/50, 34/50, 43/50, for 0, 37, 75, 150 or 300 mg/kg-bw per day, respectively) after exposure to Methyl eugenol by gavage at doses of 0, 37, 75, 150 mg/kg-bw per day, five days per week for 105 weeks or at a stop-exposure of 300 mg/kg-bw per day, five days per week for 53 weeks, followed by vehicle only for the remaining 52 weeks. The incidences of benign or malignant neuroendocrine tumours in the glandular stomach were also significantly increased in both male (0/50, 0/50, 0/50, 7/50, 4/50 for 0, 37, 75, 150 or 300 mg/kg-bw per day, respectively) and female rats (0/50, 1/50, 25/50, 34/50, 41/50 for 0, 37, 75, 150 or 300 mg/kg-bw per day, respectively). In addition, methyl eugenol significantly increased the incidences of kidney neoplasms, mammary gland fibroadenoma, malignant mesothelioma, subcutaneous fibroma or fibrosarcoma in male rats; and liver tumours hepatocholangioma or hepatocholangiocarcinoma in both male and female rats (NTP 2000a). In the carcinogenicity bioassay with B6C3F1 mice, the incidences of hepatocellular adenoma or carcinoma were increased significantly in male mice (31/50, 47/50, 46/50, 40/50, for 0, 37, 75, 150 mg/kg-bw per day, respectively) and female mice (25/50, 50/50, 49/49, 49/50, for 0, 37, 75, 150 mg/kg-bw per day, respectively) when exposed to methyl eugenol by gavage at doses of 0, 37, 75 or 150 mg/kg-bw per day, five days per week for 104 weeks. Significantly increased incidences of hepatoblastoma were also seen in female mice in a dose-related manner (NTP 2000a). The significantly increased incidences of induced liver tumours were observed at the lowest dose tested (37 mg/kg bw per day) in both rats and mice. In addition, another study showed that methyl eugenol or its metabolite 1’-hydroxyl methyl eugenol significantly induced liver tumour in mice that had received four intraperitoneal injections before weaning (total 28.3 mg/kg bw methyl eugenol or 18.3 mg/kg bw 1’-hydroxyl methyl eugenol) and were observed for up to 18 months (Miller et al 1983). Methyl eugenol was genotoxic in a number of in vivo assays in experimental animals, in vitro assays in mammalian cells and its metabolites were mutagenic in some Salmonella test strains (Appendix 3). In in vivo bioassays, methyl eugenol caused chemical-specific mutation of β-catenin gene in mouse liver tumours at 69% incidence (20/29 heptocellular neoplasms) at codons 32, 33, 34 or 41 compared with only 9% (2/22) in spontaneous tumours. Mutations in β-catenin gene caused β-catenin accumulation and up-regulation of Wnt-signaling, and subsequently stimulating cell proliferation and inhibiting apoptosis (Devereux et al 1999; NTP 2000b). In a gene mutation assay with transgenic animals, methyl eugenol significantly increased the mutation frequency of lacI gene in the liver of female Big Blue® rats administrated with Methyl eugenol by gavage at 1,000 mg/kg bw per day , 5 days/week for 90 days (8.69±3.09 ×10-5) compared with controls (1.20±0.72 × 10-5) (Tyrrell et al 2000). Moreover, in a transgenic Big Blue® male mice study, the mutation frequency of lacI in liver in methyl eugenol-treated mice (by gavage at 300 mg/kg bw per day 5 days/week for 90 days) was not significantly different from the controls. However, the mutation spectrum (pattern) from the methyl eugenol-treated group was significant different from the control group (Tyrrell et al 2000). DNA adducts were observed in the liver of CD-1 female mice treated by intraperitoneal injection of 100 or 500 mg/kg-bw methyl eugenol (Randerath et al 1984); and in the liver of newborn male B6C3F1 mice treated by intraperitoneal injection with 0.25, 0.5, 1.0, 3.0 µmol methyl eugenol on days 1, 8, 15, 22 after birth, respectively (Phillips et al 1984). Moreover both studies showed that methyl eugenol was more potent than the structurally related carcinogen safrole, in the formation of liver DNA adducts. Methyl eugenol did not induce micronuclei formation in the bone marrow of male or female B6C3F1 mice (NTP 2000a). In in vitro mammalian cell bioassays, methyl eugenol induced sister chromatid exchange in Chinese hamster ovary (CHO) cells in the presence of S9 (NTP 2000a); induced cell transformation in Syrian hamster embryo cells (Kerckaert et al. 1996); and formed DNA adducts in cultured human HepG2 cells and fibroblast V79 cells transfected with human sulphotransferase (Stening et al. 1997; Zhou et al. 2007). Methyl eugenol and its metabolite 1′-hydroxymethyl eugenol induced dose-related unscheduled DNA synthesis in cultured primary rat hepatocytes (Howes et al. 1990; Chan and Caldwell 1992). Moreover, as seen in vivo, higher DNA binding activity was observed for methyl eugenol than for the structurally related carcinogen, safrole. However, methyl eugenol did not cause chromosomal aberration in CHO cells (NTP 2000a). In bacterial bioassays, methyl eugenol was not mutagenic in Salmonella typhimurium TA97, TA98, TA100, TA102, TA1535, TA1537 or TA1538 in the presence or absence of S9; nor was it mutagenic in Escherichia coli WP2uvrA in the presence of S9 (Dorange et al. 1977; Sekizawa and Shibamoto 1982; Mortelmans et al. 1986; Schiestl et al. 1989; NTP 2000a). However, its metabolite 2′,3′-epoxymethyl eugenol induced point mutation in TA1535 and TA100 (Dorange et al. 1977); caused DNA damage in Bacillus subtilis strains M45 Rec− and H17 Rec+ (Sekizawa and Shibamoto 1982); and caused increased frequencies of intra- and interchromosomal recombination in the diploid yeast Saccharomyces cerevisiae strain RS9 (Schiestl et al. 1989) and intrachromosomal recombination in yeast strain RS112 in the presence and absence of S9 (Brennan et al. 1996). The mutagenicity, cytogenicity and DNA damage data support a conclusion that methyl eugenol is genotoxic to mammalian somatic cells in vitro and in vivo. This conclusion is consistent with the opinion of the Scientific Committee on Food on the safety of the presence of methyl eugenol in food that methyl eugenol is genotoxic and carcinogenic therefore the existence of a threshold can not be assumed (EC-SCF 2001). A fully elucidated mode of action for tumours arising as a result of exposure to methyl eugenol has not been developed. In the literature, divergent opinions have been expressed on the mode of action for such tumours. The Expert Panel of FEMA suggested that the dose-dependent hepatotoxicity is “likely to be a necessary step in carcinogenesis” in the liver (Smith et al. 2002). The induction of benign and malignant neuroendocrine tumours of the glandular stomach in rats and mice may be due in part to induction of glandular stomach atrophy, reduced gastric acid secretion, hypergastrinemia and proliferations of enterochromaffin-like cells (NTP 2000a, 2005a). However, strong evidence of mutations in β-catenin gene in mouse liver tumours led others to suggest that mutation of β-catenin may be an early event in hepatocellular tumour formation (Devereux et al. 1999; NTP 2000b; Johnson and Abdo 2005). In addition, methyl eugenol and its metabolites form DNA adducts in in vivo and in vitro assays, which suggests that DNA-reactive intermediates may also be involved in the neoplastic transformation (Johnson and Abdo 2005; NTP 2005a; Rietjens et al. 2005a, b). Burkey et al. (2000) suggested that both cytotoxicity and genotoxicity of methyl eugenol may be involved in tumour induction. The European Commission’s Scientific Committee on Food concluded that methyl eugenol is genotoxic and carcinogenic and therefore that “the existence of a threshold can not be assumed” (European Commission 2001) The mechanisms by which the alkoxy-substituted allylbenzenes, including methyl eugenol, induce cancer in experimental animals have not been established (FAO/WHO 2009). Based on the weight of evidence of carcinogenicity observed in both sexes of two experimental animal species in long-term studies, including hepatocellular carcinomas observed at the lowest test doses in these studies, and the evidence that methyl eugenol is genotoxic in a range of in vivo and in vitro assays, which included binding and damage to liver DNA and causing gene mutations in liver tumours and in transgenic animal assays, it cannot be precluded that the tumours observed in experimental animals resulted from direct interaction with genetic material. With regard to non-cancer critical effects, a significantly increased dose-related incidence of non-neoplastic lesions in the liver and glandular stomach was observed in male and female rats and mice dosed with methyl eugenol in the 2-year chronic studies. The non-neoplastic lesions in liver included eosinophilic and mixed cell foci, hepatocellular hypertrophy or hepatocyte necrosis, oval cell hyperplasia, cystic degeneration, bile duct hyperplasia, portal hypertrophy, hematopoietic cell proliferation and hemosiderin pigmentation. The non-neoplastic lesions in the glandular stomach included mucosal atrophy, neuroendocrine cell hyperplasia, glandular ectasia and chronic active inflammation. Based on the effects of the non-neoplastic lesions (hypertrophy, hyperplasia, etc.), the lowest-observed-adverse-effect level (LOAEL) was identified to be 37 mg/kg-bw per day in male and female rats and mice (NTP 2000a). In subchronic studies, cytological alteration, necrosis, hyperplasia, atrophy, and organ or body weight changes were observed in rats and mice dosed orally with methyl eugenol at doses of 0, 10, 30, 100, 300 or 1000 mg/kg-bw per day, 5 days/week for 14 weeks. Among the non-cancer critical effects, the most sensitive endpoint was reduced body weight and body weight gain, with an oral lowest-observed-effect level (LOEL) of 10 mg/kg-bw per day identified in male rats in the subchronic study (NTP 2000a; Abdo et al. 2001). In addition, intrauterine growth retardation and mildly delayed skeletal ossification were observed at the highest dose in Sprague-Dawley rats administered methyl eugenol by gavage at 0, 80, 200 or 500 mg/kg-bw per day on gestational days 6–19; however, maternal toxicity was observed at 80 mg/kg-bw per day (NTP 2004). Methyl eugenol was rapidly absorbed following oral administration to rats or mice; the plasma levels of methyl eugenol peaked within 5 minutes, and elimination of methyl eugenol from the bloodstream was rapid and multiphasic (NTP 2000a). Methyl eugenol was metabolized by the cytochrome P-450 system by three different pathways: side-chain hydroxylation, side-chain epoxidation and O-demethylation. Of the various metabolites, 1′-hydroxymethyl eugenol and methyl eugenol-2′3′-oxide were considered to be the most important ones for the toxic effects in the liver. The 1′-hydroxymethyl eugenol, followed by sulphation, subsequently formed electrophilic carbonium ions that could bind covalently to DNA and other cellular macromolecules, including proteins (Gardner et al. 1996, 1997; NTP 2005a). DNA-reactive metabolites of methyl eugenol may be a critical step involved in gene mutation and in liver tumour induction. The structurally related allylbenzene compounds, such as safrole, estragole and eugenol, are metabolized via similar pathways. A physiologically based pharmacokinetic model was developed by the NTP to represent the absorption, distribution, metabolism and elimination of methyl eugenol in rats and mice (NTP 2000a). The in vitro metabolism studies with various expressed recombinant human individual P-450 enzymes and with specific inhibition of enzymes showed that human cytochrome P-450 1A2 was the main enzyme involved in the bioactivation of methyl eugenol to 1′-hydroxymethyl eugenol and that cytochrome P-450 2C9 and 2C19 might contribute to the bioactivation at higher methyl eugenol substrate concentrations (Jeurissen et al. 2006). In vitro studies showed that human liver microsomes had comparable capacity (0.47 nmol/min/mg protein, average of 13 human liver microsomes) in bioactivation of methyl eugenol to 1-hydroxy methyl eugenol with microsomes from rats (0.42 nmal/min/mg protein, average of 25 rats from liver microsomes of all 5 test groups), although some variations were observed among humans. Taken together with other lines of evidence of bioactvation of methyl eugenol in the human liver (Jeurissen et al 2006), formation of DNA adducts in human hepatoma cells (HepG2) (Zhou et al 2007) and in human sulfotransferase-transfected fibroblast V79 cells (Stening et al 1997), all the lines of evidence suggest that Methyl eugenol can be bioactivated by human liver cells and the biologically plausibility for the human cancer risk can not be discounted. The confidence in the toxicity database for methyl eugenol is considered to be moderate. Critical effects including carcinogenicity occurred at the lowest exposure levels tested. However, the modes of action for the observed carcinogenicity of methyeugenol have not been fully elucidated. Oral dosing studies (short-term, subchronic, developmental toxicity, carcinogenicity and genotoxicity) are available. However, most of the animal studies used oral gavage as the route of administration. Repeated dose animal studies via diet, inhalation and dermal routes – the most relevant routes of human exposure - are limited or unavailable. Studies have shown that the introduction of a bolus dose of test material via gavage can lead to higher peak blood plasma levels and increased metabolic demand compared with slower more steady absorption of the substance from the diet. For example, one unpublished dietary study (Jones, 2004) was cited in the recent JECFA evaluation (2009). This study was conducted in rats with microencapsulated Methyl eugenol at dietary levels of 0,5 or 50 mg/kg bw per day over a 28 day period. No treatment-related effects were noted in the study at the highest dietary intake of 50 mg/kg bw per day. The large bolus dose delivered by oral gavage may produce metabolic and toxicological effects that are not relevant via the other routes of exposure. Characterization of Risk to Human HealthBased principally on the weight of evidence–based assessments of international or other national agencies (European Commission 2001; NTP 2005a), a critical effect for the characterization of risk to human health for methyl eugenol is carcinogenicity. Methyl eugenol is a multisite carcinogen in male and female rats and mice at all doses tested in a 2-year NTP bioassays. In the carcinogenicity studies, methyl eugenol induced multiple types of tumours in the liver and in the glandular stomach in both males and females, The liver tumours were observed at the lowest doses tested (37 mg/kg bw per day) in both rats and mice. In male rats, tumours were also observed in the kidney, mammary gland, subcutaneous tissues and mesothelium. Methyl eugenol was genotoxic in a range of in vivo and in vitro assays, although it was not mutagenic in bacterial cells. Methyl eugenol caused gene mutation in liver of transgenic animals; mutation of β-catenin gene was observed in mouse liver tumours. Modes of action have not been fully elucidated for carcinogenicity. However, based on the weight of evidence of carcinogenicity and the genotoxicity of methyl eugenol it is considered that the tumours observed in the experimental animals resulted from direct interaction with genetic material. The FAO/WHO (2009) report discussed the carcinogenic potential of methyleugenol as a single component and general population exposure to methyl eugenol as part of a larger mixture in foods or essential oils. While there is evidence to suggest that methyl eugenol is a multi-site carcinogen, there is no available data to assess the toxicological potential of the mixtures most commonly found in foods or consumer products. The FAO/WHO (2009) report further suggested that the toxicity data may not relate to the presence of methyl eugenol in natural spices based on recent in vitro data that indicates that other components of natural spices might modulate bioactivation and/or act as detoxifying agents. In the opinion of the FAO/WHO (2009) authors, the relevance of the critical effects observed in animal studies to the exposure scenario in humans was questionable and further assessment of methyleugenol was recommended. While structured epidemiological research exploring possible associations between spice consumption and hepatic cancer in humans is lacking, there is an absence of any indications of human cancer associations noted in the scientific literature. Uncertainties in Evaluation of Risk to Human HealthThe modes of tumour induction have not been fully elucidated for the various tumours observed in animal studies. While the evidence shows that DNA adducts, DNA damage and mutations play the primary role in the tumour initiation, cytotoxicity might also be involved in the tumour induction. Limited information indicates that there might be a marked variation in bioactivation of Methyl eugenol among humans. Sufficient data are not available to show the pharmacokinetic difference between animals and humans. It is assumed that the effects observed in experimental animals are relevant to humans. Liver tumours were observed at the lowest tested doses in rats and mice. In addition, there were no adequate lifetime animal studies conducted via inhalation or dermal routes of exposure. However, there is not yet any epidemiological evidence associating the natural presence of methyl eugenol in spices and spice oils, which are likely to be the main sources of methylmethyl eugenol in the diet, with liver cancer in humans. There are significant uncertainties in the estimates of dietary intake of methyl eugenol as well as in the estimate of dermal exposure to methyl eugenol from the use of personal care products. The dietary intake of methyl eugenol in Canada is difficult to estimate without detailed current data on the levels of methyl eugenol in the Canadian food supply. Spices and spice-derived essential oils are probably the primary contributors to dietary exposure to methyleugenol (based on Smith et al., 2002; WHO 2009), so the wide variation in methyl eugenol content of spices and their oils and the unknown use levels of these sources as ingredients in foods are two major factors that would lead to uncertainty in any dietary exposure assessment for methyleugenol. In the absence of Canadian data, this screening assessment presented the available dietary exposure estimates conducted internationally but a risk assessment was not done for dietary sources of methylmethyl eugenol. While restrictions on levels of methyl eugenol in essential oil ingredients in personal care products are in place in Canada , there are no data on actual concentrations in these products. Modelling of dermal exposure to methyl eugenol did not account for all types of product that may be formulated with oils containing methyl eugenol, nor did the estimate account for market share of individual products within a product category. ConclusionBased on the information presented in this draft screening assessment, it is proposed that Methyl eugenol is not entering the environment in a quantity or concentration or under conditions that have or may have an immediate or long-term harmful effect on the environment or its biological diversity or that constitute or may constitute a danger to the environment on which life depends. Additionally, the substance meets the criteria for persistence but not the criteria for bioaccumulation potential as set out in the Persistence and Bioaccumulation Regulations (Canada 2000). 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Ottawa (ON): Health Canada, Pest Management Regulatory Agency. [cited 2009 Aug]. Available from: http://www.hc-sc.gc.ca/cps-spc/pest/part/consultations/_pacr2004-36/index-eng.php Health Canada. 2007. The cosmetic ingredient hotlist – March 2007 [Internet]. Ottawa (ON): Health Canada, Consumer Product Safety. [cited 2009 Aug]. Available from: http://www.hc-sc.gc.ca/cps-spc/person/cosmet/info-ind-prof/_hot-list-critique/hotlist-liste_e.html Health Canada. 2008. Review of the 2004 re-evaluation of citronella oil and related active compounds for use as personal insect repellents. Re-evaluation Note REV2008-03. Health Canada. 2009. An Act to amend the Tobacco Act [Internet]. Ottawa (ON): Health Canada. [cited 2009 Aug]. Available from: http://www.hc-sc.gc.ca/hc-ps/tobac-tabac/legislation/federal/amend_faq-modif-eng.php [HENRYWIN] Henry’s Law Constant Program for Microsoft Windows [Estimation Model]. 2008 [please check year: I think this version may have been issued in 2007]. Version 3.20. 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Unpublished report to the Flavor and Extract Manufacturers Association, Washington, DC, USA. Submitted to the World Health Organization by the International Organization of the Flavour Industry, Brussels, Belgium. [cited in FAO/WHO 2009]. Keith LH. 1976. Identification of organic compounds in unbleached treated kraft paper mill wastewaters. Environ Sci Technol 10(6):555–564. Kerckaert GA, Brauninger R, LeBoeuf RA, Isfort RJ. 1996. Use of the Syrian hamster embryo cell transformation assay for carcinogenicity prediction of chemicals currently being tested by the National Toxicology Program in rodent bioassays. Environ Health Perspect 104:1075–1084. [KOWWIN] Octanol–Water Partition Coefficient Program for Microsoft Windows [Estimation Model]. 2008. Version 1.67. Washington (DC): US Environmental Protection Agency, Office of Pollution Prevention and Toxics; Syracuse (NY): Syracuse Research Corporation. [cited 2009 Oct]. Available from: http://www.epa.gov/oppt/exposure/pubs/episuite.htm Larsen W, Nakayama H, Fischer T, Elsner P, Frosch P, Burrows D, Jordan W, Shaw S, Wilkinson J, Marks J, Sugawara M, Nethercott M, Nethercott J. 2002. Fragrance contact dermatitis—a worldwide multicenter investigation (Part III). Contact Dermatitis 46:141–144. Lewis RJ Sr. 2001. Hawley’s condensed chemical dictionary. 14th ed. New York (NY): John Wiley & Sons. p. 735. Lide DR, Milne GWA, editors. 1994. Handbook of data on organic compounds, vol. I. 3rd ed. Boca Raton (FL): CRC Press. p. 867. [LNHPD] Licensed Natural Health Products Database [database on the Internet]. 2009. Ottawa (ON): Health Canada. [cited 2009 Oct]. Available from: http://www.hc-sc.gc.ca/dhp-mps/prodnatur/applications/licen-prod/lnhpd-bdpsnh-eng.php Mekenyan G, Dimitrov SD, Pavlov TS, Veith GD. 2005. POPs: a QSAR system for creating PBT profiles of chemicals and their metabolites. SAR QSAR Environ Res 16(1–2):103–133. Merck. 1997. Safety data sheet. 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Mortelmans K, Haworth S, Lawlor T, Speck W, Tainer B, Zeiger E. 1986. Salmonella mutagenicity tests. II. Results from the testing of 270 chemicals. Environ Mutagen 8:1–119. [NCI] National Chemical Inventories [database on a CD-ROM]. 2006. Columbus (OH): American Chemical Society. [cited 2009 Oct]. Available from: http://www.cas.org/products/cd/nci/index.html [NTP] National Toxicology Program (US). 2000a. Toxicology and carcinogenesis studies of methyl eugenol (CAS No. 93-15-2) in F344/N rats and B6C3F1 mice (gavage studies). Research Triangle Park (NC): US Department of Health and Human Services, National Toxicology Program. NTP Technical Report 491. NIH Publication No. 00-3950. Available from: http://ntp.niehs.nih.gov/ntp/htdocs/LT_rpts/tr491.pdf [NTP] National Toxicology Program (US). 2000b. Final report on carcinogens. Background document for methyleugenol (CAS No. 93-15-2). December 13–14, 2000. Meeting of the NTP Board of Scientific Counselors, Report on Carcinogens Subcommittee. Available from: http://ntp.niehs.nih.gov/ntp/newhomeroc/roc10/ME.pdf [NTP] National Toxicology Program (US). 2004. Final study report of the developmental toxicity evaluation for methyl eugenol (CAS No. 93-15-2) administered by gavage to Sprague-Dawley (CD) rats on gestational days 6 through 19. Research Triangle Park (NC): US Department of Health and Human Services, National Toxicology Program. NTP Study TER97007. [Full report available from National Technical Information Service as NTIS No. PB2004-104348]. [NTP] National Toxicology Program (US). 2005a. Substance profile: methyl eugenol (CAS No. 93-15-2). In: Report on carcinogens. 11th ed. Research Triangle Park (NC): US Department of Health and Human Services, National Toxicology Program. [cited 2009 Sep 28]. Available from: http://ntp.niehs.nih.gov/ntp/roc/eleventh/profiles/s109meth.pdf [NTP] National Toxicology Program (US). 2005b. Substance profile: safrole (CAS No. 94-59-7). In: Report on carcinogens. 11th ed. 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Mutat Res 224(4):427–436. Sekizawa J, Shibamoto T. 1982. Genotoxicity of safrole-related chemicals in microbial test systems. Mutat Res 101:127–140. Shaver TN, Bull DL. 1980. Environmental fate of methyl eugenol. Bull Environ Contam Toxicol 24:619–626. Smith RL, Adams TB, Doull J, Feron VJ, Goodman JI, Marnett LJ, Portoghese PS, Waddell WJ, Wagner BM, Rogers AE, Caldwell J, Sipes IG. 2002. Safety assessment of allylalkoxybenzene derivatives used as flavouring substances—methyl eugenol and estragole. Food Chem Toxicol 40:851–870. Snell ML, Abdo KM, Herbert RA, Eldridge S, Cummingham ML. 2000. Subchronic toxicity of methyl eugenol administered by gavage to F-344 rats and B6C3F1 mice. Toxicologist 54:267 (abstract). Stanfill SB, Calafat AM, Brown CR, Polzin GM, Chiang JM, Watson CH, Ashley DL. 2003. Concentrations of nine alkenylbenzenes, coumarin, piperonal and pulegone in Indian bidi cigarette tobacco. Food Chem Toxicol 41(2):303–317. Stening P, Gardner I, Kenna JE, Coughtrie MWH, Caldwell J. 1997. Formation of alkenylbenzene macromolecular adducts in human fibroblast V79 cells transfected with human sulfotransferases. Human Exp Toxicol 16:62 (abstract). Turner B, Miller N, Tran D, Powell S. 1989. The environmental monitoring of methyl eugenol, naled and dichlorvos during a pest trapping and eradication program. Sacramento (CA): California Department of Food and Agriculture. 48 p. Tyrrell SP, Zhang X, Cunningham ML, Shane BS. 2000. Comparison of the mutagenicity of methyl eugenol (ME) in vivo in the liver of Big Blue® transgenic rats and mice. Toxicologist 54:229 (abstract). [US EPA] US Environmental Protection Agency. 2006. Methyl eugenol (ME) (203900) fact sheet [Internet]. Washington (DC): US EPA. [cited 2009 Aug]. Available from: http://www.epa.gov/pesticides/biopesticides/ingredients/factsheets/factsheet_203900.htm [US EPA] US Environmental Protection Agency. 2008. Screening-level hazard characterization of high production volume chemicals. Sponsored chemical: estragole (CAS No. 140-67-0). Prepared by: US EPA, Risk Assessment Division, High Production Volume Chemicals Branch, Risk Assessment Division, United States Environmental Protection Agency [US FDA] US Food and Drug Administration. 2001. 21 CFR §172.515. Synthetic flavoring substances and adjuvants [Internet]. Washington (DC): US Department of Health and Human Services. [cited 2009 Aug]. Available from: http://edocket.access.gpo.gov/cfr_2001/aprqtr/pdf/21cfr172.515.pdf Zhou GD, Moorthy B, Bi J, Donnelly KC, Randerath K. 2007. DNA adducts from alkoxyallylbenzene herb and spice constituents in cultured human (HepG2) cells. Environ Mol Mutagen 48(9):715–721. Appendix I - Robust Study Summary
Appendix 2. Various references re: Naturally occurring methyl eugenol content of essential oils. (adapted from Burfield 2004))
Aurore et al.: Aurore GS, Abaul J, Bourgeois P, Luc J. 1998. Antibacterial and antifungal activities of the essential oils of Pimenta racemosa var. racemosa P. Miller (J.W. Moore) (Myrtaceae). J Essential Oil Res 10(2):161–164.
Appendix 3ConsExpo 4.1 report Product Body Lotion 0.005% Methyl eugenol
Output
Appendix 4. Summary of health effects information for methyl eugenol
[1] A determination of whether one or more of the criteria of section 64 are met is based upon an assessment of potential risks to the environment and/or to human health associated with exposures in the general environment. For humans, this includes, but is not limited to, exposures from ambient and indoor air, drinking water, foodstuffs, and the use of consumer products. A conclusion under CEPA 1999 on the substances in the Chemicals Management Plan (CMP) Challenge Batches 1-12 is not relevant to, nor does it preclude, an assessment against the hazard criteria specified in the Controlled Products Regulations, which is part of regulatory framework for the Workplace Hazardous Materials Information System [WHMIS] for products intended for workplace use. Disclaimer: Although care has been taken to ensure that the information found on this website accurately reflects the requirements prescribed in the Canadian Environmental Protection Act (1999), you are advised that, should any inconsistencies be found, the legal documents, printed in the Canada Gazette, will prevail. |
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