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EC number: 941-212-1 | CAS number: -
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Endpoint summary
Administrative data
Key value for chemical safety assessment
Effects on fertility
Description of key information
- The absence of evidence of reproductive and developmental toxicity in an already extensive dataset.
- The control of risks to human health in all identified exposure scenarios
- changes in thyroid and uterus weight (in terms of weight or change relative to total body weight)
- gross pathological abnormalities at necropsy in exposed animals including the uterus
- microscopic observations in the thyroid gland
- Developmental endpoints by dose for litters with implants, namely:
- number of corpora luteum
- number of implantation site
- number of live and dead foetuses
- number of early and late resorptions
- number of late resorptions
- number of pre- and post-implantation losses
- Developmental endpoints by dose for litters with live foetuses, namely:
- number of live offspring
- foetal sex ratio
- foetal body weights
- anogenital distance of foetuses
- any of the developmental parameters investigated as part of the external, visceral and skeletal examinations for malformations and variations.
- Hand protection to EN374 standard as minimum and Eye protection to EN166 standard as minimum are employed for all operations.
- Protective clothing to EN368 standard as minimum is used to restrict exposure to the workforce.
- There is a definitive body of evidence that the test substance does not adversely affect the reproductive processes of mating and pregnancy and does not affect the numbers born to exposed females and the early development of the offspring. A series of reliable and relevant sub-acute and sub-chronic (OECD TG414 and TG408) studies have been conducted which provide detailed and extensive data on a range of reproductive and developmental endpoints. The OECD TG414 Prenatal Development Toxicity Study indicates that exposure to the test substance Distillation Residue Grade resulted in no adverse effects on the reproductive organs and fertility of parental animals and the development of resulting offspring. In an OECD TG408 Repeated Dose 90-day Toxicity Study there were no observable changes on reproductive organs/tissues in males and females compared to those from the control animals at the highest test dose.
- Although the conduct of the EOGRTS test could potentially provide additional data on reproductive and developmental toxicity endpoints it would not improve the overall conclusions on the perceived risks posed to human health by exposure to the test substance via the identified exposure scenarios in the Chemical Safety Report. In addition, studies show that general systemic toxic effects only occur in the presence of local effects due to the known irritancy of the test substance. Therefore, sustained dermal exposure which could potentially lead to systemic effects in workers and consumers is not to be expected without significant local effects. This would limit the total dose to which individuals would be exposed.
- Taking account of all these points together it is proposed to adapt the conduct of the Extended One-Generation Reproductive Toxicity Study on Distilled Grade and adopt a weight of evidence approach. Given the currently available dataset addressing key reproduction and developmental toxicity endpoints conduct of the EOGRTS would be disproportionate and inappropriate for this Natural Complex Substance.
Weight of evidence for Section 8.7.3 Extended One-Generation Reproductive Toxicity Study (EOGRTS) of Annex X of the REACH Regulation
Requirement for an Extended One-Generation Reproductive Toxicity Study (EOGRTS)
The Extended One-Generation Reproductive Toxicity Study (EOGRTS) (OECD TG443, EU B.56) in its basic test design is a standard information requirement at REACH Annex IX (where concerns regarding reproductive toxicity are evident) and X levels. EOGRTS was introduced under Commission Regulation (EU) 2015/282 of 20 February 2015 amending Annexes VIII, IX and X to the REACH Regulation (EC No. 1907/2006). It is a test method developed to assess the reproductive toxicity of chemical substances and is the preferred test method to address the standard information requirement defined in Column 1 of Point 8.7.3 of Annexes IX and X to the REACH Regulation instead of the Two-Generation Reproductive Toxicity study. Standard information requirement for reproduction toxicity related to EOGRTS in Annexes IX and X is limited to the basic configuration of EOGRTS, which includes only cohorts 1A and IB without extension to include a F2 generation and developmental neurotoxicity and developmental immunotoxicity cohorts.
There are no Specific Rules for Adaptation from Column 1 in Annex X Section 8.7 of the REACH Regulation, but these are in place for the test at Annex IX. However, adaptions pursuant to Annex XI (General Rules for Adaptation of the Standard Testing Regime set out in Annexes VII to X) and a recent Judgement of the General Court in Case T-755/17 (September 2019) in relation to the Principle of Proportionality are considered relevant to the potential for adaption of this test requirement in appropriate instances. Paragraph 287 of the Judgement states that:
“The relevant criterion relating to the principle of proportionality is the result of balancing the different objectives pursued by Regulation No 1907/2006 and the application of the precautionary principle. In accordance with that criterion, in order to justify a request to conduct testing, the ECHA must not only demonstrate the existence of a potential risk for human health and the environment, and the necessity to clarify that risk, but also establish that there is a realistic likelihood that the information requested would allow improved risk management measures to be taken”.
The requirement to conduct complex mammalian toxicity tests at Annex IX which potentially involves the use of hundreds of vertebrates (normally rats) needs to be carefully considered to ensure that it satisfies Article 25(1) of the REACH Regulation that any testing on vertebrate animals is undertaken only as a last resort. Furthermore, any testing carried out needs to satisfy Recital 63 of the REACH Regulation which sets out a general principle according to which ‘[i]t is also necessary to ensure that generation of information is tailored to real information needs…’. Where testing is proposed it is also important to ensure that in the studies conducted the fewest number of animals possible are used to satisfy the objective pursued as required by Directive 2010/63/EU of the European Parliament and of the Council on the protection of animals used for scientific purposes (OJ L 276, 20.10.2010, p. 33). Furthermore, the extent to which the test information will improve the quantification and management of risks to human health and the environment based on the Risk Characterisation Exercise needs to be demonstrated. Therefore, an implicit presumption underlying the requirement to provide data from an EOGRTS study on Distillation Residue Grade is that without the requested information it will not be possible to verify whether there remains an uncontrolled risk with the substance that should be subject to further risk management measures.
The OECD TG443 Test Guideline indicates that “the main objective of the Extended One-Generation Reproductive Toxicity Study is to evaluate specific life stages not covered by other types of toxicity studies and test for effects that may occur as a result of pre- and postnatal chemical exposure”. With regard to other related toxicity studies the Test Guideline states that “For reproductive endpoints, it is envisaged that, as a first step and when available, information from repeat-dose studies (including screening reproductive toxicity studies, e.g. TG 422 (32)), or short term endocrine disrupter screening assays, (e.g. Uterotrophic assay - TG 440 (36); and Hershberger assay - TG 441 (37)) is used to detect effects on reproductive organs for males and females. This might include spermatogenesis (testicular histopathology) for males and oestrous cycles, follicle counts/oocyte maturation and ovarian integrity (histopathology) for females”
In its basic design EOGRTS provides an evaluation of the pre- and postnatal effects of chemicals on development as well as a thorough evaluation of systemic toxicity in pregnant and lactating females and young and adult offspring. The test chemical is administered continuously (on a once a day basis) in graduated doses to several groups of sexually-mature males and females. This parental (P) generation is dosed for a defined pre-mating period (selected based on the available information for the test chemical, but for a minimum of two weeks) and a two-week mating period. Parental generation males are further treated at least until weaning of the F1 offspring, which involves treatment for a minimum of 10 weeks. They may be treated for longer if there is a need to clarify effects on reproduction. Treatment of the P females is continued during pregnancy and lactation until termination after the weaning of their litters (i.e. after 8-10 weeks of treatment). The F1 offspring receive further treatment with the test chemical from weaning to adulthood. In the basic study design, detailed examination of key developmental endpoints, such as offspring viability, neonatal health, developmental status at birth, and physical and functional development until adulthood, is expected to identify specific target organs in the offspring. In addition, the study will provide and/or confirm information about the effects of a test chemical on the integrity and performance of the adult male and female reproductive systems. Specifically, but not exclusively, the following parameters are considered: gonadal function, the oestrous cycle, epididymal sperm maturation, mating behaviour, conception, pregnancy, parturition, and lactation.
REACH Annex XI (General Rules for Adaptation of the Standard Testing Regime set out in Annexes VII to X) indicates that there are a number of approaches by which a registrant may adapt the standard testing regime in accordance with the general rules set out in Section 1 of the Annex. Section 1.2 relates to the use of a weight of evidence approach and states that “There may be sufficient weight of evidence from several independent sources of information leading to the assumption/conclusion that a substance has or has not a particular dangerous property, while the information from each single source alone is regarded insufficient to support this notion”. This approach has been used to address the data requirement for the Extended One-Generation Reproductive Toxicity Study (EOGRTS) for Distillation Residue Grade at Annex X.
Although there is no defined rationale for adapting the EOGRTS test given in Annex X this is not the case for Annex IX. This Annex allows for the possibility of not conducting the basic text design and Column 1 of Section 8.7.3 states that the test is required “if the available repeated dose toxicity studies (e.g. 28- or 90-days studies or OECD TGs 421 or 422 screening tests) indicate adverse effects on reproductive organs or tissues or reveal other concerns in relation with reproductive toxicity. Information from non-animal approaches are thus not listed as triggers for this study at REACH Annex IX level in the REACH Annex text. However, if there is a serious concern based on available information from non-animal approaches or structurally analogous substances, the study may be triggered” (ECHA, 2017). The inference from this tonnage-based information strategy is that there could be no circumstances at Annex X where the test would not need to be conducted and the test should be triggered irrespective of the existing evidence regarding the extent of the effects of the test substance on reproductive and developmental endpoints. However, this is contrary to the view expressed in Recital 63 of the REACH Regulation and the Judgement of the General Court in Case T-755/17. This should mean that the test does not need to be carried out if there is sufficient reliable data available to assess the potential hazards and risks posed by the substance to pregnant females and their offspring and provide a weight of evidence that further testing is not required as the potential risks are adequately controlled.
This principle has been confirmed by an ECHA Board of Appeal ruling on 25 September 2018 in Case A-008-2017. This annulled the Agency’s decision of 23 March 2017 on the substance evaluation of 2,2',6,6'-tetra-tert-butyl-4,4'-methylenediphenol pursuant to Article 46 of REACH. The Appellants claimed that the Agency failed to fulfil the conditions for imposing further information requirements, including the need for EOGRTS, under Article 46 of the REACH Regulation and failed to provide an adequate statement of reasons, in particular regarding the potential risk perceived by the Agency. In addition, the Appellants claimed that the Agency failed to provide information or explanations on how the information required in the Contested Decision would lead to an improvement of the risk management measures in place.
In their judgement the Board of Appeal upheld the Appellant’s view that the Agency had made no evaluation whatsoever of whether the available information showed 'serious concerns about the potential for adverse effects on fertility or development' within the meaning of Column 2 of Section 8.7.1. of Annex VIII of the REACH Regulation. Although this judgement relates to a substance at a lower tonnage band than that relevant to Distillation Residue Grade and relates to substance evaluation rather than compliance checking it establishes principles requiring ECHA to evaluate the existing data to demonstrate that the conduct of an EOGRTS can be justified to further assess the perceived risks from a test substance, particularly given the large numbers of mammals that would be used. This weight of evidence document reviews the available relevant mammalian reproductive and developmental data for Distillation Residue Grade and provides arguments for not conducting the EOGRTS.
Weight of evidence approach for the EOGRTS
The weight of evidence approach for addressing the EOGRTS endpoint for Distilled Grade is based on two elements:
Information relevant to each of these elements is provided in the following sections.
Absence of evidence of reproductive and developmental toxicity in an already extensive dataset
Currently reliable and relevant data is available on the potential reproductive and developmental toxicity of Distillation Residue Grade to rats from a Prenatal Developmental Toxicity Study (OECD TG414). A Repeated Dose 90-day Toxicity Study (OECD TG408) is also available which provides reliable data on the potential effects of Distillation Residue Grade on the growth and maturation of male and female rats. It is considered that on the basis of this dataset the EOGRTS study is not necessary to provide further clarification on whether the test substance constitutes a risk to human health and the environment. Given the currently available dataset addressing key reproduction and developmental toxicity endpoints conduct of the EOGRTS would be disproportionate and inappropriate for Distillation Residue Grade.
It should be noted that the dermal route of exposure is the most relevant in terms of potential human exposure to Distillation Residue Grade rather than the oral route based on normal use patterns. However, Distillation Residue Grade is classified as a Category 2 Skin Irritant (H315) and it was not considered appropriate from an animal welfare viewpoint to conduct the study using this route of exposure. Therefore, all the existing data assessing reproductive and developmental toxicity has been generated in tests using the oral route.
Reliable and relevant data (Klimisch Code 1) are available from a Prenatal Developmental Toxicity Study (OECD TG414) on Distillation Residue Grade which assessed the effects of test substance exposure on offspring birth and development. The test started with the arrival of groups of pre-mated female Wistar rats. Untreated females were mated at the organism supplier and were at Day 0 or 1 post-coitum on arrival (Day 0 post-coitum is the day of successful mating). Twenty-two time-mated females (of approximately 10 to 14 weeks age) were assigned to a control group (no exposure to test substance) and three test substance exposure groups (i.e. doses of 100, 300 and 1000 mg/kg bw/day). The test substance and vehicle (propylene glycol) were administered to the appropriate animals by once daily oral gavage 7 days a week from Day 6 to Day 21 post-coitum, inclusive, which corresponds to the female pregnancy and foetal in-utero development phases of the EOGRTS test. Clinical signs, functional observations, body weight development and food and water consumption were monitored during the study along with an extensive series of developmental endpoints in the foetuses. These included litter size, post-implantation loss, foetal sex ratio, foetal body weights, foetal anogenital distance and external, visceral and skeletal malformations and developmental variations in foetuses.
In the study no statistically significant treatment-related maternal effects were evident up to 1000 mg/kg bw/day in terms of:
In the study no statistically significant treatment-related effects were also observed for:
In summary, the data from the OECD TG414 study indicates that no maternal systemic toxicity and no developmental toxicity was observed in the 100, 300 and 1000 mg/kg/day groups, as no relevant endpoints showed significant differences in responses compared to the control animals. No effects on endocrine-disruption related apical morphological and histological endpoints (i.e. anogenital distance and sex ratio in foetuses, thyroid weight and histopathology in females) and indicators of thyroid hormonal activity, i.e. Triiodothyronine (T3), Thyroxine (T4) and Thyroid Stimulating Hormone (TSH) levels, in females were observed in the OECD TG414 study on Distillation Residue Grade. Based on the results in this prenatal developmental toxicity study a maternal and developmental No Observed Adverse Effect Level (NOAEL) for (Distillation Residue Grade) of 1000 mg/kg/day was established.
An assessment of test substance exposure on the growth and maturation of post-weaning rats through to adulthood are available from a reliable and relevant data (Klimisch Code 1) Repeated Dose 90-day Toxicity Study (OECD TG408) on Distillation Residue Grade. In the test, groups of ten male and ten female (nulliparous and non-pregnant) Wistar rats (approximately 6 weeks of age[1]) were allocated to a control group (no test substance) and three test substance exposure groups (i.e. doses of 100, 300 and 1000 mg/kg bw/day). The test substance and vehicle (propylene glycol) were administered to the appropriate animals by once daily oral gavage 7 days a week for a minimum of 90 days, up to and including the day before scheduled necropsy. Clinical signs, functional observations, body weight development and food and water consumption were monitored during the study. In the study the additional sperm parameters measured in the EOGRTS were not addressed and the vaginal smears collected were only assessed at necropsy to determine the stage of the oestrous cycle and allow correlation with the histopathology of ovaries.
At end of treatment no changes in the weights of a large number of organs were evident including the male and female reproductive organs (i.e. epididymis, seminal vesicle gland and testis in males and the ovary and uterus in females). Higher liver weights were noted for males treated at 300 and 1000 mg/kg bw/day and females treated at 300 (relative to body weight only) and 1000 mg/kg bw/day (absolute and relative to body weight). This increased liver weight correlated to hepatocellular hypertrophy and an increased incidence of multinucleated hepatocytes, which were seen microscopically in males and females at 1000 mg/kg bw/day and males at 300 mg/kg bw/day. Additionally, clinical biochemistry measurements revealed concurrent changes in liver enzymes. A >4-fold increase in alanine aminotransferase activity (ALAT) was noted in males and females treated at 1000 mg/kg bw/day, in combination with lesser increases in aspartate aminotransferase (ASAT) and alkaline phosphatase (ALP) activity and total bilirubin concentration, which suggests hepatocellular damage. Therefore, the changes in the liver at 1000 mg/kg bw/day are considered adverse. The liver changes at 300 mg/kg bw/day were, at the severity observed, not considered to be adverse.
During microscopic examination, erosion/ulcers were noted in the stomach of males at 1000 mg/kg/day. Due to the degenerative nature of this finding, the effect was considered to be adverse and was considered to be due to the known irritancy of the test substance.
Histopathological examination of a large number of organs/tissues in males and females showed that test substance-related microscopic findings after treatment were only noted in the liver (as described previously), mesenteric lymph node, spleen and thyroid glands se well as the stomach of males of males and females. In general these effects were observed at the highest dose (1000 mg/kg bw/day), but were typically of limited severity. With regard to the effects on the thyroid gland, males, starting at the mid dose of 300 mg/kg bw/day, showed an increased incidence and severity (compared to concurrent control males) of diffuse, bilateral, follicular cell hypertrophy, up to mild degree in the highest dose of 1000 mg/kg bw/day. Given the largely minimal severity of the response (except for a mild degree of severity observed in some males at the highest dose), which was without any degenerative changes, these findings were not considered to be adverse. In females a low incidence and minimal severity of follicular cell hypertrophy was noted in treated animals. The effect in females was also without any degenerative changes so the findings were not considered to be adverse. In the study, a marked increase of TSH was observed in males at 300 and 1000 mg/kg bw/day and females at 1000 mg/kg bw/day. However, there was marked variability in the responses of individuals at the highest exposure groups and most values were within the 95%ile for the historical control data. There was no accompanying change in thyroid weight in either males or females. Under the conditions of this study no adverse effect was observed that could be linked to the increase in TSH and thyroid follicular cell hypertrophy. No test-substance induced changes in T3 and T4 levels were recorded in males and females during the study. Overall, it is concluded that no adverse effects on endocrine-disruption related apical morphological and histological endpoints and indicators of thyroid hormonal activity in both sexes were observed in the OECD TG408 study on Distillation Residue Grade. Based on the results observed in the stomach and liver, the No Observed-Adverse-Effect Level (NOAEL) for local and systemic toxicity was considered to be 300 mg/kg bw/day.
In summary, reliable and relevant sub-acute and sub-chronic (OECD TG414 and TG408) studies have been conducted which provide detailed and extensive data on a range of reproductive and developmental endpoints. The OECD TG414 study indicates that exposure to the test substance Distillation Residue Grade resulted in no adverse effects on the reproductive organs and fertility of parental animals and the development of resulting offspring. In an OECD TG408 Repeated Dose 90-day Toxicity Study there were no observable changes on reproductive organs/tissues in males and females compared to those from the control animals at the highest test dose.
it is accepted that not all the parameters relating to reproductive and developmental toxicity measured in the EOGRTS have been addressed in the OECD TG414 and TG408 studies. However, given the absence of adverse effects in the currently available dataset addressing key reproduction and developmental toxicity endpoints for Distillation Residue Grade the conduct of the EOGRTS would be disproportionate and inappropriate for this Natural Complex Substance. The data for Distillation Residue Grade is supported by a corresponding dataset for OECD TG414 and TG408 studies (as well as an OECD TG422 study) on another category member Distilled Grade.
Control of risks to human health in all exposure scenarios
The Risk Characterisation Exercise in the Chemical Safety Report (CSR) carried out using the updated Derived No Effect Levels (DNELs) generated using the new data from Annex IX and X testing with the OECD TG408 and TG414 studies shows that in all cases the RCR values for human health (workers and consumers) assessment in all the identified exposure scenarios were less than 1(indicating an acceptable level of risk as humans are not exposed to the test substance at levels above the relevant “safe” threshold(s)). .
Distillation Residue Grade is classified as a Skin Sensitiser Cat 1A, Skin Irritant Cat. 2 and Eye Damage Cat 1. Given the classification of Distillation Residue Grade as a skin irritant and sensitiser and an eye irritant all skin and mucous membranes with potential exposure should be protected with appropriate PPE. Therefore, in all the exposure scenarios appropriate risk management measures in the CSR are specified, namely:
On this basis workers are expected to be protected adequately from exposure (according to controlled risk described in the CSR) and thus systemic effects after dermal exposure are expected to be minimal. For consumers it is assumed that the advice provided on personal protection during the use of products containing Distillation Residue Grade are followed.
In addition, the OECD TG414 and TG408 studies show that general systemic toxic effects only occur in the presence of local effects due to the known irritancy of the constituents of the test substance. Therefore, sustained dermal exposure which could potentially lead to systemic reproductive and developmental effects in workers and consumers is not to be expected without significant local effects which would restrict exposure.
Overall, it is not expected that provision of further information from the EOGRTS would change the perceived risk posed to the general population (consumers) and workers by exposure to Distillation Residue Grade via the exposure scenarios identified in the Chemical Safety Report.
Summary
The following points are considered relevant to the adaption of the Extended One Generation Reproductive Toxicity Study for Distillation Residue Grade:
References
ECHA (2017) Guidance on Information Requirements and Chemical Safety Assessment. Chapter R.7a: Endpoint specific guidance (Version 6.0, July 2017). European Chemical Agency, Helsinki.
ECHA Board of Appeal (2018) Judgement Case Number A-008-2017, Summary of Decision of 25th September 2018 of the Board of Appeal of the European Chemicals Agency. Details available at: https://echa.europa.eu/documents/10162/6fcc6c62-3e36-3aa8-e213-38289b84a3e4
Klimisch, H-J, Andreae, M and Tillmann, U (1997) A systematic approach for evaluating the quality of experimental toxicological and ecotoxicological data. Regulatory Toxicology and Pharmacology, 25, 1–5.
Judgement of the General Court (2019) In Case T-755/17. Details available at: http://curia.europa.eu/juris/celex.jsf?celex=62017TJ0755&lang1=en&type=TXT&ancre
[1] In the OECD TG408 study dosing should begin as soon as possible after weaning and, in all cases, before the animals are nine weeks old
Link to relevant study records
- Endpoint:
- screening for reproductive / developmental toxicity
- Type of information:
- read-across based on grouping of substances (category approach)
- Adequacy of study:
- key study
- Justification for type of information:
- 1. HYPOTHESIS FOR THE CATEGORY APPROACH (ENDPOINT LEVEL)
The basis for the adoption of a category approach for the three grades of processed Cashew Nutshell Extract is the commonality of the constituents and functional groups in the three grades and the common modes of action for specific localised endpoints that are manifest in physico-chemical, environmental fate and toxicological properties that are similar or follow a regular pattern as a result of structural similarity. The ECHA Final Decisions on the Annex IX and X (mammalian toxicology, ecotoxicology and environmental fate) Testing Proposals for three grades confirmed the applicability of a category approach which involves reading data across from the source substances Cashew Nutshell Extract, Decarboxylated, Distilled (Distilled Grade) and Cashew Nutshell Extract, Decarboxylated, Distillation Residue (Distillation Residue Grade) to the target substance Cashew Nutshell Extract, Decarboxylated (Technical Grade) by interpolation. In this context interpolation is “the estimation of a value for a member of the group using measured values from other members on both sides of that member within the defined group spectrum”. Further details on the justification for using the interpolation based read-across approach are given in the attached document “Report on the grouping and read-across rationale for the three grades of processed Cashew Nutshell Extract”. The interpolation approach has been applied to the Annex IX OECD TG408 and TG414 testing on Distilled and Distillation Residue Grades. For the Annex VII and VIII endpoints a programme of testing has been carried out to provide comparative information for the three grades and develop a data matrix to support the category and read-across approaches. This has involved generating reliable data for all Annex VII and VIII physico-chemical and environmental fate endpoints and certain mammalian toxicity properties. However, based on the discussions surrounding the Final Decisions it was not considered necessary to apply this approach retrospectively to address all identified data gaps at Annexes VII and VIII. The original data used for the registration of a single substance (Cashew Nutshell Liquid, CAS Number 8007-24-7) was largely based on data for Distilled Grade since this was considered to be the most (eco) toxicologically active form, given the higher content of low molecular weight constituents (such as cardanol) and the low content of polymeric species. Therefore, based on the use of the category approach it is considered appropriate to read-across from the existing short-term mammalian toxicity data for Distilled Grade to the other two grades.
2. CATEGORY APPROACH JUSTIFICATION (ENDPOINT LEVEL)
For the mammalian reproductive toxicity endpoint no test data is available for the registered substance Distillation Residue Grade. However data is available for the compositionally similar substance Distilled Grade which is part of the category of the three grades of CNSL. In a subacute 28-day Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test Distilled Grade was administered to 5 Sprague-Dawley rats/sex/dose via oral gavage at dose levels of 0, 15, 150, or 1000 mg/kg bw/day). Changes in the lungs, mesenteric lymph node occurred at the highest dose of 1000 mg/kg bw/day resulting in a NOAEL of 150 mg/kg bw/day for systemic toxicity. A NOAEL of 1000 mg/kg bw/day has been identified for reproductive toxicity based on an absence of effects of the test substance on the relevant reproductive parameters. It is considered appropriate to read-across from the data for Distilled Grade to Distillation Residue Grade with the result that it has been estimated that Distillation Residue Grade does not result effects on mammalian reproductive toxicity parameters at an exposure dose of 1000 mg/kg bw/day. - Clinical signs:
- effects observed, treatment-related
- Body weight and weight changes:
- effects observed, treatment-related
- Food consumption and compound intake (if feeding study):
- effects observed, treatment-related
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Histopathological findings: non-neoplastic:
- no effects observed
- Other effects:
- no effects observed
- Reproductive function: oestrous cycle:
- not examined
- Reproductive function: sperm measures:
- not examined
- Reproductive performance:
- no effects observed
- Dose descriptor:
- NOAEL
- Effect level:
- 150 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: BAsed on systemic changes in the lungs, mesenteric lymph node, stomach and duodenum.
- Clinical signs:
- no effects observed
- Mortality / viability:
- no mortality observed
- Body weight and weight changes:
- no effects observed
- Sexual maturation:
- not examined
- Organ weight findings including organ / body weight ratios:
- not examined
- Gross pathological findings:
- no effects observed
- Histopathological findings:
- not examined
- Dose descriptor:
- NOAEL
- Generation:
- F1
- Effect level:
- 1 000 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: No treatment effects occurred on reproduction or offspring development.
- Reproductive effects observed:
- not specified
- Conclusions:
- Changes in the lungs, mesenteric lymph node occurred at the highest dose of 1000 mg/kg bw/day resulting in a NOAEL of 150 mg/kg bw/day for systemic toxicity. A NOAEL of 1000 mg/kg bw/day has been identified for reproductive toxicity based on an absence of effects of the test substance on the relevant reproductive parameters. It is considered appropriate to read-across from the data for Distilled Grade to Distillation Residue Grade with the result that it has been estimated that Distillation Residue Grade does not result effects on mammalian reproductive toxicity parameters at an exposure dose of 1000 mg/kg bw/day.
- Executive summary:
In a subacute 28-day Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test Distilled Grade was administered to 5 Sprague-Dawley rats/sex/dose via oral gavage at dose levels of 0, 15, 150, or 1000 mg/kg bw/day). Changes in the lungs, mesenteric lymph node occurred at the highest dose of 1000 mg/kg bw/day resulting in a NOAEL of 150 mg/kg bw/day for systemic toxicity. A NOAEL of 1000 mg/kg bw/day has been identified for reproductive toxicity based on an absence of effects of the test substance on the relevant reproductive parameters. It is considered appropriate to read-across from the data for Distilled Grade to Distillation Residue Grade with the result that it has been estimated that Distillation Residue Grade does not result effects on mammalian reproductive toxicity parameters at an exposure dose of 1000 mg/kg bw/day.
- Endpoint:
- extended one-generation reproductive toxicity - basic test design (Cohorts 1A, and 1B without extension)
- Type of information:
- read-across based on grouping of substances (category approach)
- Adequacy of study:
- weight of evidence
- Study period:
- 2016
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study
- Justification for type of information:
- 1. HYPOTHESIS FOR THE CATEGORY APPROACH (ENDPOINT LEVEL)
The basis for the adoption of a category approach for the three grades of processed Cashew Nutshell Extract is the commonality of the constituents and functional groups in the three grades and the common modes of action for specific localised endpoints that are manifest in physico-chemical, environmental fate and toxicological properties that are similar or follow a regular pattern as a result of structural similarity. The ECHA Final Decisions on the Annex IX and X (mammalian toxicology, ecotoxicology and environmental fate) Testing Proposals for three grades confirmed the applicability of a category approach which involves reading data across from the source substances Cashew Nutshell Extract, Decarboxylated, Distilled (Distilled Grade) and Cashew Nutshell Extract, Decarboxylated, Distillation Residue (Distillation Residue Grade) to the target substance Cashew Nutshell Extract, Decarboxylated (Technical Grade) by interpolation. In this context interpolation is “the estimation of a value for a member of the group using measured values from other members on both sides of that member within the defined group spectrum”. Further details on the justification for using the interpolation based read-across approach are given in the attached document “Report on the grouping and read-across rationale for the three grades of processed Cashew Nutshell Extract”. The interpolation approach has been applied to the Annex IX OECD TG408 and TG414 testing on Distilled and Distillation Residue Grades. For the Annex VII and VIII endpoints a programme of testing has been carried out to provide comparative information for the three grades and develop a data matrix to support the category and read-across approaches. This has involved generating reliable data for all Annex VII and VIII physico-chemical and environmental fate endpoints and certain mammalian toxicity properties. However, based on the discussions surrounding the Final Decisions it was not considered necessary to apply this approach retrospectively to address all identified data gaps at Annexes VII and VIII. The original data used for the registration of a single substance (Cashew Nutshell Liquid, CAS Number 8007-24-7) was largely based on data for Distilled Grade since this was considered to be the most (eco) toxicologically active form, given the higher content of low molecular weight constituents (such as cardanol) and the low content of polymeric species. Therefore, based on the use of the category approach it is considered appropriate to read-across from the existing short-term mammalian toxicity data for Distilled Grade to the other two grades.
2. CATEGORY APPROACH JUSTIFICATION (ENDPOINT LEVEL
For the mammalian reproductive toxicity endpoint limited non-rodent test data is available for the registered substance, Distillation Residue Grade. However, data on the reproductivity of cattle is available for the compositionally similar substance Technical Grade which is part of the category of the three grades of CNSL. A series of studies by Kaneda and Mochizuk (2016), which are classified as reliable with restriction (Klimisch Code 2), have investigated the potential to which feeding cattle with pellets containing unprocessed Cashew Nutshell Extract and Technical Grade results in changes in reproductivity as well as changes in milk yield and/or milk quality and the incidence of perinatal disease (which accounts for more than half the deaths in cattle and results in the disposal of huge quantities of milk). The studies have been. The test results from the reproductivity study indicate that the feeding of the Technical Grade-containing pellet was found to result in improvements in almost all the endpoints for the test group compared with control group. On this basis Technical Grade does not evidently have any adverse toxic effects on the reproduction of cattle at the dose tested. It is considered appropriate to read-across from the data for Technical Grade to Distillation Residue Grade as part of a weight of evidence approach for the EOGRTS information requirement. - GLP compliance:
- not specified
- Dose descriptor:
- NOAEL
- Effect level:
- 20 mg/kg bw/day
- Based on:
- test mat.
- Sex:
- female
- Basis for effect level:
- reproductive performance
- Dose descriptor:
- dose level:
- Generation:
- F1
- Effect level:
- ca. 10 other: g/day
- Based on:
- test mat.
- Sex:
- not specified
- Remarks on result:
- not determinable due to absence of adverse toxic effects
- Reproductive effects observed:
- no
- Conclusions:
- Data is available on the potential effects on the reproductivity of cattle of the compositionally similar substance Technical Grade (which is part of the category of the three grades of CNSL). A series of studies by Kaneda and Mochizuk (2016), which are classified as reliable with restriction (Klimisch Code 2), have investigated the potential to which feeding cattle with pellets containing unprocessed Cashew Nutshell Extract and Technical Grade (which is compositionally similar to Distillation Residue Grade) results in changes in reproductivity as well as changes in milk yield and/or milk quality and the incidence of perinatal disease (which accounts for more than half the deaths in cattle and results in the disposal of huge quantities of milk). The test results from the reproductivity study indicate that the feeding of the Technical Grade-containing pellet was found to result in improvements in almost all the endpoints for the test group compared with control group. On this basis Technical Grade does not evidently have any adverse toxic effects on the reproduction of cattle at the dose tested.
- Executive summary:
In the study evaluating the effect of feeding Technical Grade on reproductivity, adult female Holstein cattle were used with the mature cows (i.e. those having had at least one calf) weighing between 450 kg (1,000 pounds) and 680 kg (1,500 pounds) depending on their age. Individuals which had calved were divided at random into a test group and control group. Technical Grade was fed to the individuals in the test group in pellets at an amount of 100 g (10 g in terms of CNSL) per day for 5 days from the date of calving. The feeding rate is equivalent to approximately 20 mg/kg bw/day of CNSL. The Technical Grade used largely comprised 55-80% cardanol and 5-30% cardol, which is consistent with the substance identity profile of the registered substance. The test group, which were fed Technical Grade, consisted of 9 cows (2 cows having had two calves, 4 cows having had three calves, 1 cow having had four calves, 1 cow having had five calves and 1 cow having had six calves). Five cows (all of which had had three calves) were allocated to control group and were not fed Technical Grade containing-pellets.
The test results from the reproductivity study indicate that the feeding of the Technical Grade-containing pellet was found to result in improvements in almost all the endpoints for the test group compared with control group. In particular, significant effects in terms of reducing the number of inseminations, decreasing the number of non-pregnant days and actual non-pregnant days and improving the pregnancy rate were measured. Further, the feeding of Technical grade was able to achieve a conception rate exceeding 50-60% and an oestrus detection rate exceeding 70% or more of target levels, respectively. No change was noted in the days of first insemination. Overall, the results show that the feeding of the Technical Grade-containing pellet was found to significantly improve the reproductivity in each individual cow and to further significantly improve the reproductivity in the oestrus detection rate and pregnancy rate in each group. On the basis of this data Technical Grade does not evidently have any adverse toxic effects on the reproduction of cattle at the dose tested.
- Endpoint:
- extended one-generation reproductive toxicity - basic test design (Cohorts 1A, and 1B without extension)
- Type of information:
- read-across based on grouping of substances (category approach)
- Adequacy of study:
- weight of evidence
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Justification for type of information:
- 1. HYPOTHESIS FOR THE CATEGORY APPROACH (ENDPOINT LEVEL)
The basis for the adoption of a category approach for the three grades of processed Cashew Nutshell Extract is the commonality of the constituents and functional groups in the three grades and the common modes of action for specific localised endpoints that are manifest in physico-chemical, environmental fate and toxicological properties that are similar or follow a regular pattern as a result of structural similarity. The ECHA Final Decisions on the Annex IX and X (mammalian toxicology, ecotoxicology and environmental fate) Testing Proposals for three grades confirmed the applicability of a category approach which involves reading data across from the source substances Cashew Nutshell Extract, Decarboxylated, Distilled (Distilled Grade) and Cashew Nutshell Extract, Decarboxylated, Distillation Residue (Distillation Residue Grade) to the target substance Cashew Nutshell Extract, Decarboxylated (Technical Grade) by interpolation. In this context interpolation is “the estimation of a value for a member of the group using measured values from other members on both sides of that member within the defined group spectrum”. Further details on the justification for using the interpolation based read-across approach are given in the attached document “Report on the grouping and read-across rationale for the three grades of processed Cashew Nutshell Extract”. The interpolation approach has been applied to the Annex IX OECD TG408 and TG414 testing on Distilled and Distillation Residue Grades. For the Annex VII and VIII endpoints a programme of testing has been carried out to provide comparative information for the three grades and develop a data matrix to support the category and read-across approaches. This has involved generating reliable data for all Annex VII and VIII physico-chemical and environmental fate endpoints and certain mammalian toxicity properties. However, based on the discussions surrounding the Final Decisions it was not considered necessary to apply this approach retrospectively to address all identified data gaps at Annexes VII and VIII. The original data used for the registration of a single substance (Cashew Nutshell Liquid, CAS Number 8007-24-7) was largely based on data for Distilled Grade since this was considered to be the most (eco) toxicologically active form, given the higher content of low molecular weight constituents (such as cardanol) and the low content of polymeric species. Therefore, based on the use of the category approach it is considered appropriate to read-across from the existing short-term mammalian toxicity data for Distilled Grade to the other two grades.
2. CATEGORY APPROACH JUSTIFICATION (ENDPOINT LEVEL)
For the mammalian reproductive toxicity endpoint data on offspring birth and development are also available for the compositionally similar substance Distilled Grade which is part of the category of the three grades of CNSL. In a reliable and relevant (Klimisch Code 1) OECD TG414 Prenatal Developmental Toxicity Study, Distilled Grade was administered to twenty-two time-mated females (of approximately 10 to 14 weeks age) assigned to a control group (no exposure to test substance) and three test substance exposure groups (i.e. doses of 100, 300 and 1000 mg/kg bw/day). The test substance and vehicle were administered to the appropriate animals by once daily oral gavage 7 days a week from Day 6 to Day 21 post-coitum, inclusive. Clinical signs, functional observations, body weight development and food and water consumption were monitored during the study along with an extensive series of developmental endpoints in the foetuses.
In the study animals in the high dose treatment (1000 mg/kg bw/day) group experienced severe toxicity (as body weight loss, decreased food consumption and adverse clinical signs) over Days 14 to 17. To prevent further suffering, it was decided by the veterinarian at the test laboratory to sacrifice all the remaining high dose (1000 mg/kg/day) animals on Study Day 18 for ethical reasons. It was concluded that the effects observed at the highest dose were probably due to the known irritancy of the test substance which caused localised changes in stomach morphology and limited food consumption. No mortality was observed for the animals in the remaining groups during the study.
The test substance administered at dose levels of 100 mg/kg bw/day was not considered to cause maternal toxicity. No systemic toxic effects and non-adverse local effects on the surface of the forestomach were observed. At the 300 mg/kg bw/day dose level, the test substance was maternally toxic causing both adverse systemic toxic effects (on body weight gain). Body weight gain at 300 mg/kg bw/day was slightly lower from Day 12 post-coitum onwards, reaching statistical significance on Days 18 and 21 post-coitum only (0.88x and 0.83x of controls, respectively). Body weight gain, corrected for the weight of the uterus of the pregnant female, was statistically significantly lower compared with concurrent controls (on average 50% lower). Adverse local effects on the surface of the forestomach were also evident at 300 mg/kg bw/day which could also have caused secondary effects on body weight gain.
No toxicologically significant changes were noted in any of the developmental parameters after treatment up to 300 mg/kg bw/day. Due to the premature sacrifice of the dams treated at 1000 mg/kg bw/day, no data on foetal parameters were determined at this dose level. However, in the 1000 mg/kg bw/day group all except two females were gravid on the day of unscheduled necropsy.
The data from the OECD TG414 study indicates that general systemic toxic effects (in terms of body weight gain) were occurring at the highest exposure dose (300 mg/kg bw/day) where animals survived to necropsy whilst potentially endocrine-mediated effects on reproductive and developmental endpoints were not evident at any treatment dose (100 and 300 mg/kg bw/day). No effects on endocrine-disruption related apical morphological and histological endpoints (i.e. anogenital distance and sex ratio in foetuses, thyroid weight and histopathology in females) and indicators of thyroid hormonal activity (i.e. T3, T4 and TSH levels) in females were observed in the OECD TG414 study on Distilled Grade. It is considered appropriate to read-across from the data for Distilled Grade to Distillation Residue Grade as part of a weight of evidence approach for the EOGRTS information requirement since the OECD TG414 studies on both the source substances show an absence of effects on reproductive and developmental endpoints at the highest dose tested in animals surviving to necropsy. - Species:
- rat
- Strain:
- Wistar
- Dose descriptor:
- NOAEL
- Effect level:
- 100 mg/kg bw/day
- Based on:
- test mat.
- Sex:
- female
- Basis for effect level:
- other: maternal toxicity
- Remarks on result:
- not measured/tested
- Reproductive effects observed:
- no
- Conclusions:
- In the absence of adverse findings in any of the other parameters at the lowest dose level, the systemic maternal No Observed Adverse Effect Level (NOAEL) for Cashew Nutshell Extract, Decarboxylated, Distilled (Distilled Grade) was determined to be 100 mg/kg bw/day.
The developmental NOAEL for Distilled Grade was established as being at least 300 mg/kg bw/day, as no litters were available at 1000 mg/kg bw/day as these dams were sacrificed in extremis - Executive summary:
Reliable and relevant data (Klimisch Code 1) are also available from a Prenatal Developmental Toxicity Study (OECD TG414) on Distilled Grade which assessed the effects of test substance exposure on offspring birth and development. The test started with the arrival of groups of pre-mated female Wistar rats. Untreated females were mated at the organism supplier and were at Day 0 or 1 post-coitum on arrival (Day 0 post-coitum is the day of successful mating). Twenty-two time-mated females (of approximately 10 to 14 weeks age) were assigned to a control group (no exposure to test substance) and three test substance exposure groups (i.e. doses of 100, 300 and 1000 mg/kg bw/day). The test substance and vehicle (propylene glycol) were administered to the appropriate animals by once daily oral gavage 7 days a week from Day 6 to Day 21 post-coitum, inclusive, which corresponds to the female pregnancy and foetal in-utero development phases of the EOGRTS test.
No toxicologically significant changes were noted in any of the developmental parameters after treatment up to 300 mg/kg bw/day. Due to the premature sacrifice of the dams treated at 1000 mg/kg bw/day, no data on foetal parameters were determined at this dose level. However, in the 1000 mg/kg bw/day group all except two females were gravid on the day of unscheduled necropsy.
The data from the OECD TG414 study indicates that general systemic toxic effects (in terms of body weight gain) were occurring at the highest exposure dose (300 mg/kg bw/day) where animals survived to necropsy whilst potentially endocrine-mediated effects on reproductive and developmental endpoints were not evident at any treatment dose (100 and 300 mg/kg bw/day). No effects on endocrine-disruption related apical morphological and histological endpoints (i.e. anogenital distance and sex ratio in foetuses, thyroid weight and histopathology in females) and indicators of thyroid hormonal activity (i.e. T3, T4 and TSH levels) in females were observed in the OECD TG414 study on Distilled Grade.
- Endpoint:
- extended one-generation reproductive toxicity - basic test design (Cohorts 1A, and 1B without extension)
- Type of information:
- read-across based on grouping of substances (category approach)
- Adequacy of study:
- weight of evidence
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Justification for type of information:
- 1. HYPOTHESIS FOR THE CATEGORY APPROACH (ENDPOINT LEVEL)
The basis for the adoption of a category approach for the three grades of processed Cashew Nutshell Extract is the commonality of the constituents and functional groups in the three grades and the common modes of action for specific localised endpoints that are manifest in physico-chemical, environmental fate and toxicological properties that are similar or follow a regular pattern as a result of structural similarity. The ECHA Final Decisions on the Annex IX and X (mammalian toxicology, ecotoxicology and environmental fate) Testing Proposals for three grades confirmed the applicability of a category approach which involves reading data across from the source substances Cashew Nutshell Extract, Decarboxylated, Distilled (Distilled Grade) and Cashew Nutshell Extract, Decarboxylated, Distillation Residue (Distillation Residue Grade) to the target substance Cashew Nutshell Extract, Decarboxylated (Technical Grade) by interpolation. In this context interpolation is “the estimation of a value for a member of the group using measured values from other members on both sides of that member within the defined group spectrum”. Further details on the justification for using the interpolation based read-across approach are given in the attached document “Report on the grouping and read-across rationale for the three grades of processed Cashew Nutshell Extract”. The interpolation approach has been applied to the Annex IX OECD TG408 and TG414 testing on Distilled and Distillation Residue Grades. For the Annex VII and VIII endpoints a programme of testing has been carried out to provide comparative information for the three grades and develop a data matrix to support the category and read-across approaches. This has involved generating reliable data for all Annex VII and VIII physico-chemical and environmental fate endpoints and certain mammalian toxicity properties. However, based on the discussions surrounding the Final Decisions it was not considered necessary to apply this approach retrospectively to address all identified data gaps at Annexes VII and VIII. The original data used for the registration of a single substance (Cashew Nutshell Liquid, CAS Number 8007-24-7) was largely based on data for Distilled Grade since this was considered to be the most (eco) toxicologically active form, given the higher content of low molecular weight constituents (such as cardanol) and the low content of polymeric species. Therefore, based on the use of the category approach it is considered appropriate to read-across from the existing short-term mammalian toxicity data for Distilled Grade to the other two grades.
2. CATEGORY APPROACH JUSTIFICATION (ENDPOINT LEVEL)
For the mammalian reproductive toxicity endpoint limited non-rodent test data is available for the registered substance, Distillation Residue Grade. However, data on the growth and maturation of post-weaning rats through to adulthood is also available for the compositionally similar substance Distilled Grade which is part of the category of the three grades of CNSL. In a reliable and relevant (Klimisch Code 1) sub-chronic OECD TG408 Repeated Dose 90-day Toxicity Study, Distilled Grade was administered to groups of ten male and ten female (nulliparous and non-pregnant) Wistar rats (approximately 6 weeks of age) allocated to a control group (no test substance) and three test substance exposure groups (i.e. doses of 50, 1500 and 500 mg/kg bw/day). The test substance and vehicle were administered to the appropriate animals by once daily oral gavage 7 days a week for a minimum of 90 days, up to and including the day before scheduled necropsy. Clinical signs, functional observations, body weight development and food and water consumption were monitored during the study.
Exposure to Distilled Grade did not result in adverse effects in rats at levels of 50 and 150 mg/kg bw/day. Histopathological examinations showed epithelial hyperplasia of the non-glandular mucosa of the stomach (forestomach, squamous mucosa) at 50, 150 and 500 mg/kg bw/day in males and females with a dose-related increase in severity, up to marked in males and moderate in females. These findings at 500 mg/kg bw/day were interpreted to be adverse due to the overall severity and concurrent presence of ulceration in some animals. Furthermore, a lower body weight gain was observed in males at 500 mg/kg bw/day which was also considered to be adverse. Other test substance-related changes in clinical signs, body weight, clinical pathology, organ weights and histopathology were considered non-adverse. This included an assessment of the absolute weights of a series of reproductive organs/tissues (testes, epididymides, prostate gland and seminal vesicle in males and ovary and uterus in females) which showed that there were no changes, relative to those in the controls, at any exposure dose. However, when the results were adjusted relative to the terminal body weights of the rats, small, but statistically significant, increases in the weight of the testis (20%) and the epididymis (18%), relative to body weight, were evident in males exposed to 500 mg/kg bw/day when compared to measurements made on the controls. Histopathological examination of the organs/tissues in males and females showed that none of the reproductive organs/tissues in males and females showed observable changes compared to those from the control animals indicating that prolonged exposure did not affect the male and female reproductive systems.
In the study, a marked increase of TSH was observed in males at 150 and 500 mg/kg bw/day and females at 500 mg/kg bw/day, though there was marked variability with most values being within the 95%ile limit for historical control data . This change correlated with diverse follicular cell hypertrophy in the thyroid of males, which were without degenerative changes. Under the conditions of this study no adverse effect was observed that could be linked to the increase in TSH and thyroid follicular cell hypertrophy. Overall no adverse effects on endocrine-disruption related apical morphological and histological endpoints and indicators of thyroid hormonal activity in both sexes were observed in the OECD TG408 study on Distilled Grade.
It is considered appropriate to read-across from the data for Distilled Grade to Distillation Residue Grade as part of a weight of evidence approach for the EOGRTS information requirement since the OECD TG408 studies on both the source substances showed that prolonged exposure did not affect the male and female reproductive systems in rats. - Species:
- rat
- Strain:
- Wistar
- Dose descriptor:
- NOAEL
- Remarks:
- Systemic toxicity
- Effect level:
- 150 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- organ weights and organ / body weight ratios
- Dose descriptor:
- NOAEL
- Remarks:
- Local effects
- Effect level:
- 500 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- histopathology: non-neoplastic
- Remarks on result:
- not measured/tested
- Reproductive effects observed:
- no
- Conclusions:
- Administration of Cashew Nutshell Extract, Decarboxylated, Distilled (Distilled Grade) by once daily oral gavage for 90 days did not result in adverse effects in rats at levels of 50 and 150 mg/kg bw/day. Histopathological examinations showed epithelial hyperplasia of the non-glandular mucosa of the stomach (forestomach, squamous mucosa) at 50, 150 and 500 mg/kg bw/day in males and females with a dose-related increase in severity, up to marked in males and moderate in females. At 150 and 500 mg/kg bw/day this correlated with macroscopically irregular stomach surfaces. These findings at 500 mg/kg bw/day were interpreted to be adverse due to the overall severity and concurrent presence of ulceration in some animals. Furthermore, a lower body weight gain was observed in males at 500 mg/kg bw/day which was also considered to be adverse.
Other test substance-related changes in clinical signs, body weight, clinical pathology, organ weights and histopathology were considered non-adverse.
Based on the results generated a No Observed Adverse Effect Level (NOAEL) for the systemic toxicity of Cashew Nutshell Extract, Decarboxylated, Distilled (Distilled Grade) of 150 mg/kg bw/day was established. This NOAEL also provides protection against the observed local effects on stomach morphology due to the known irritancy of the test substance. - Executive summary:
An assessment of test substance exposure on the growth and maturation of post-weaning rats through to adulthood is available from a reliable and relevant data (Klimisch Code 1) Repeated Dose 90-day Toxicity Study (OECD TG408) on Distilled Grade. In the test, groups of ten male and ten female (nulliparous and non-pregnant) Wistar rats (approximately 6 weeks of age) were allocated to a control group (no test substance) and three test substance exposure groups (i.e. doses of 50, 150 and 500 mg/kg bw/day). The test substance and vehicle (propylene glycol) were administered to the appropriate animals by once daily oral gavage 7 days a week for a minimum of 90 days, up to and including the day before scheduled necropsy.
Exposure to Distilled Grade did not result in adverse effects in rats at levels of 50 and 150 mg/kg bw/day. Histopathological examinations showed epithelial hyperplasia of the non-glandular mucosa of the stomach (forestomach, squamous mucosa) at 50, 150 and 500 mg/kg bw/day in males and females with a dose-related increase in severity, up to marked in males and moderate in females. At 150 and 500 mg/kg bw/day this correlated with macroscopically irregular stomach surfaces. These findings at 500 mg/kg bw/day were interpreted to be adverse due to the overall severity and concurrent presence of ulceration in some animals. Furthermore, a lower body weight gain was observed in males at 500 mg/kg bw/day which was also considered to be adverse. Other test substance-related changes in clinical signs, body weight, clinical pathology, organ weights and histopathology were considered non-adverse. This included an assessment of the absolute weights of a series of reproductive organs/tissues (testes, epididymides, prostate gland and seminal vesicle in males and ovary and uterus in females) which showed that there were no changes, relative to those in the controls, at any exposure dose. However, when the results were adjusted relative to the terminal body weights of the rats, small, but statistically significant, increases in the weight of the testis (20%) and the epididymis (18%), relative to body weight, were evident in males exposed to 500 mg/kg bw/day when compared to measurements made on the controls. Histopathological examination of the organs/tissues in males and females showed that none of the reproductive organs/tissues in males and females showed observable changes compared to those from the control animals indicating that prolonged exposure did not affect the male and female reproductive systems.
In the study, a marked increase of TSH was observed in males at 150 and 500 mg/kg bw/day and females at 500 mg/kg bw/day, though there was marked variability between the responses of the individuals in these groups with most values being within the 95%ile limit for historical control data. This change correlated with diverse follicular cell hypertrophy in the thyroid of males, which were without degenerative changes. Under the conditions of this study no adverse effect was observed that could be linked to the increase in TSH and thyroid follicular cell hypertrophy. Overall no adverse effects on endocrine-disruption related apical morphological and histological endpoints and indicators of thyroid hormonal activity in both sexes were observed in the OECD TG408 study on Distilled Grade.
Referenceopen allclose all
There were no toxicologically significant deaths during the study.
Increased salivation was detected prior to dosing and up to 5 hours after dosing for animals of either sex treated with 1000 mg/kg bw/day from Day 9 onwards. One female treated with 150 mg/kg bw/day developed clinical signs consistent with inappropriate dosing on Day 5 and was subsequently terminated. One female treated with 1000 mg/kg bw/day had given birth to a number of pups of which the majority were found dead. Several clinical signs were observed in this animal, and the animal and litter were terminated.
BODY WEIGHT AND WEIGHT GAIN
A slightly reduced bodyweight gain was observed for 1000 mg/kg bw/day males during the first two weeks of the study. Reduced bodyweight gain was also observed for 1000 mg/kg bw/day females during the later stages of the gestation period.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
No adverse effect on dietary intake or food efficiency were detected.
FOOD EFFICIENCY
No adverse effect on dietary intake or food efficiency were detected.
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study)
No intergroup differences were detected.
OPHTHALMOSCOPIC EXAMINATION
N/A
HAEMATOLOGY
Haematological assessments revealed elevated platelet counts in animals of either sex treated with 1000 mg/kg bw/day. Elevated haemoglobin, erythrocyte and haematocrit was also evident for males treated at 1000 mg/kg bw/day. No such effects were detected at 150 and 15 mg/kg bw/day.
CLINICAL CHEMISTRY
An increase in aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase levels were observed for 1000 mg/kg bw/day animals of either sex, together with elevated inorganic phosphorus, bilirubin and urea, and reduced cholesterol levels.
URINALYSIS
N/A
NEUROBEHAVIOUR
Open field arena observations revealed increased salivation for individual animals of either sex treated with 1000 mg/kg bw/day from Week 3.
ORGAN WEIGHTS
Females treated with 1000 mg/kg bw/day showed elevated liver weights.
GROSS PATHOLOGY
None
HISTOPATHOLOGY: NON-NEOPLASTIC
Groups of alveolar macrophages were seen with a higher incidence for females treated with 1000 mg/kg bw/day.
A higher incidence of sinus histiocytosis and/or foamy histiocytes was observed in relation to treatment with 1000 mg/kg bw/day.
Hyperkeratosis, frequently associated with acanthosis was seen in the forestomachs of animals of either sex treated with 1000 mg/kg bw/day. Focal ulceration of the forestomach epithelium was also seen in one high dose female.
Mucosal hypertrophy was seen in three males at the top dose group. A low incidence of mucosal hypertrophy was observed in females in all dose groups.
OTHER FINDINGS
MATING
No adverse effects on mating or fertility were observed.
OFFSPRING LITTER SIZE AND VIABILITY
No effects detected.
OFFSPRING GROWTH AND DEVELOPMENT
No effects detected.
LITTER OBSERVATIONS
No effects detected.
UTERINE EXAMINATION
No effects detected.
No effects detected.
OFFSPRING GROWTH AND DEVELOPMENT
No effects detected.
LITTER OBSERVATIONS
No effects detected.
UTERINE EXAMINATION
No effects detected.
Effects of feeding Technical grade on the reproductivity of cows
Endpoint |
Mean results for the Technical grade group (n = 9)
|
Mean results for the control group (n = 5) |
Variation |
Number of insemination times |
2.0 |
2.6 |
-0.6 |
Non-pregnant days |
111 |
137 |
-26 |
Non-pregnant days: 90 days of less (%) |
33.3 |
20.0 |
13.3 |
Non-pregnant days: 120 days of more (%) |
33.3 |
60.0 |
-26.7 |
Actual non-pregnant days |
32.1 |
58.3 |
-26.2 |
Days of first insemination |
78.9 |
78.7 |
0.2 |
Pregnancy rate at first service (%) |
33.3 |
16.7 |
16.6 |
Pregnancy rate (%) |
50.0 |
38.5 |
11.5 |
Oestrus detection rate (%) |
79.1 |
68.9 |
10.2 |
Notes:
Number of insemination = times of artificial insemination
Non-pregnant days = last conception day - last calving day
Actual non-pregnant days = days from first to last insemination
Days of first insemination = first insemination day after calving - last calving day
Pregnancy rate at first service = rate of conception achieved by first insemination
Pregnancy rate = [number of individuals with conception (group total)/total times of artificial insemination] x 100
Detection rate of oestrus (whole group) = [average number of insemination/((average actual days of non-pregnant/21)+1] x100
Effect on fertility: via oral route
- Endpoint conclusion:
- no adverse effect observed
- Dose descriptor:
- NOAEL
- 1 000 mg/kg bw/day
Effect on fertility: via inhalation route
- Endpoint conclusion:
- no study available
Effect on fertility: via dermal route
- Endpoint conclusion:
- no study available
Additional information
For the assessment of the toxicity to reproduction to Distillation Residue Grade additional information is available for the read across substance is Cashew (Anacardium occidentale) Nutshell Extract, Decarboxylated, Distilled (Distilled Grade). A document which justifies the read–across from Cashew (Anacardium occidentale) Nutshell Extract, Decarboxylated, Distilled (Distilled Grade) to Cashew (Anacardium occidentale) Nutshell Extract, Decarboxylated, Distillation Residue (Distillation Residue Grade) is attached.
Short description of key information:
In a subchronic toxicity study (OECD TG414) Cashew Nutshell Extract, Decarboxylated, Distillation Residue (Distillation Residue Grade) was administered to Wistar Han rats at dose levels of 0, 150, 300, or 1000 mg/kg bw/day). No toxicologically significant changes at exposure levels up to 1000 mg/kg bw/day were noted in any of the remaining maternal parameters investigated in this study (i.e. mortality/moribundity, clinical appearance, body weight, food consumption, gross pathology, thyroid weights, uterine contents, histopathologic examination (thyroid gland), corpora lutea, implantation sites and pre- and post- implantation loss). No test substance-related changes at exposure levels up to 1000 mg/kg bw/day were noted in any of the developmental parameters investigated in this study (i.e. litter size, sex ratio, fetal body weights, fetal anogenital distance and external, visceral and skeletal malformations and variations in fetuses). In the absence of adverse findings at the highest dose level, the systemic maternal No Observed Adverse Effect Level (NOAEL) for Cashew Nutshell Extract, Decarboxylated, Distillation Residue (Distillation Residue Grade) was determined to be 1000 mg/kg bw/day.
The developmental NOAEL for Distillation Residue Grade was also established as being 1000 mg/kg bw/day.
In an additional subchronic toxicity study (OECD TG422) on a read-across substance Cashew Nutshell Extract, Decarboxylated, Distilled (Distilled Grade) was administered to 5 Sprague-Dawley rats/sex/dose via gavage at dose levels of 0, 15, 150, or 1000 mg/kg bw/day). Systemic changes in the lungs, mesenteric lymph node, stomach and duodenum occurred at the highest dose. The NOAEL is 150 mg/kg bw/day. A NOAEL of 1000 mg/kg bw/day has been identified for reproductive parameters. This repeat dose/reproductive toxicity screening study in the rat is acceptable and satisfies the guideline requirement for a subchronic oral study OECD 422 in rats.
Effects on developmental toxicity
Description of key information
In a subchronic toxicity study (OECD TG414) Cashew Nutshell Extract, Decarboxylated, Distillation Residue (Distillation Residue Grade) was administered to Wistar Han rats at dose levels of 0, 150, 300, or 1000 mg/kg bw/day). No toxicologically significant changes at exposure levels up to 1000 mg/kg bw/day were noted in any of the remaining maternal parameters investigated in this study (i.e. mortality/moribundity, clinical appearance, body weight, food consumption, gross pathology, thyroid weights, uterine contents, histopathologic examination (thyroid gland), corpora lutea, implantation sites and pre- and post- implantation loss). No test substance-related changes at exposure levels up to 1000 mg/kg bw/day were noted in any of the developmental parameters investigated in this study (i.e. litter size, sex ratio, fetal body weights, fetal anogenital distance and external, visceral and skeletal malformations and variations in fetuses). In the absence of adverse findings at the highest dose level, the systemic maternal No Observed Adverse Effect Level (NOAEL) for Cashew Nutshell Extract, Decarboxylated, Distillation Residue (Distillation Residue Grade) was determined to be 1000 mg/kg bw/day.
The developmental NOAEL for Distillation Residue Grade was also established as being 1000 mg/kg bw/day.
In an addition a subchronic toxicity study on a read-across substance Cashew Nutshell Extract, Decarboxylated, Distilled (Distilled Grade) was administered to 5 Sprague-Dawley rats/sex/dose via gavage at dose levels of 0, 15, 150, or 1000 mg/kg bw/day). Systemic changes in the lungs, mesenteric lymph node, stomach and duodenum occurred at the highest dose. The NOAEL is 150 mg/kg bw/day. A NOAEL of 1000 mg/kg bw/day has been identified for reproductive parameters. This repeat dose/reproductive toxicity screening study in the rat is acceptable and satisfies the guideline requirement for a subchronic oral study OECD 422 in rats.
Link to relevant study records
- Endpoint:
- developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 18th August to 5th September 2019
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Remarks:
- under GLP
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 414 (Prenatal Developmental Toxicity Study)
- Version / remarks:
- OECD TG414 dated June 2018
- Deviations:
- no
- Remarks:
- No deviations were considered to have impacted the overall integrity of the study or the interpretation of the study results and conclusions.
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
- Species:
- rat
- Strain:
- Wistar
- Remarks:
- Hannover
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: From Charles River Laboratories Deutschland, Sulzfeld, Germany
- Age at study initiation: At the initiation of dosing females were 10 to 14 weeks old
- Weight at study initiation: At the initiation of dosing females were between 186 and 274 g
- Fasting period before study:
- Housing: Individually in Macrolon plastic cages (MIII type, height 18 cm) containing appropriate bedding (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany)
- Diet (e.g. ad libitum): Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) was provided ad libitum throughout the study, except during designated procedures
- Water (e.g. ad libitum): Municipal tap water was freely available to each animal via water bottles
- Acclimation period: The animals were allowed to acclimate to the Test Facility toxicology accommodation for at least 5 days before the commencement of dosing.
ENVIRONMENTAL CONDITIONS
- Temperature (°C): Target temperatures of 18 to 24°C with the actual daily mean temperature during the study period being 21°C.
- Humidity (%): Relative target humidity of 40 to 70% with the actual daily mean relative humidity being 49 to 73%.
- Air changes (per hr): Ten or greater air changes per hour with 100% fresh air (no air recirculation) were maintained in the animal rooms
- Photoperiod (hrs dark / hrs light): A 12 hour light/12 hour dark cycle was maintained
IN-LIFE DATES: From: To: - Route of administration:
- oral: gavage
- Vehicle:
- corn oil
- Remarks:
- Dried and Deacidified
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS: Test substance dosing formulations (w/w) were homogenized to visually acceptable levels at appropriate concentrations to meet dose level requirements. The dosing formulations were prepared weekly as a solution, filled out in daily portions and stored in the refrigerator protected from light. The dosing formulations were removed from the refrigerator and stirred at room temperature for at least 30 minutes before dosing.
DIET PREPARATION
- Rate of preparation of diet (frequency):
- Mixing appropriate amounts with (Type of food):
- Storage temperature of food:
VEHICLE
- Justification for use and choice of vehicle (if other than water):
- Concentration in vehicle:
- Amount of vehicle (if gavage):
- Lot/batch no. (if required):
- Purity: - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- Homogeneity/stability analyses of dosing solutions were performed at each dose level prior to the initiation of the study.
The concentrations analyzed in the formulations of the 100 mg/kg/day (low dose), 300 mg/kg bw/day (mid dose) and 1000 mg/kg bw/day (high dose) were in agreement with target concentrations (i.e. mean accuracies between 85% and 115%). - Details on mating procedure:
- The females arrived on Day 0 or Day 1 post-coitum (Day 0 post-coitum is defined as the day of successful mating).
- Duration of treatment / exposure:
- Days 6 - 20 post-coitum inclusive
- Frequency of treatment:
- Females only, Once/day. Treated from Day 6 through 20 post-coitum inclusive
- Duration of test:
- 20 days
- Dose / conc.:
- 0 mg/kg bw/day
- Remarks:
- Actual ingested
- Dose / conc.:
- 100 mg/kg bw/day
- Remarks:
- Actual ingested
- Dose / conc.:
- 300 mg/kg bw/day
- Remarks:
- Actual ingested
- Dose / conc.:
- 1 000 mg/kg bw/day
- Remarks:
- Actual ingested
- No. of animals per sex per dose:
- 22 females
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- A total of 88 pregnant Wistar Han female rats were allocated to four groups (22 per group) Three groups were administered with the test substance Distilled Grade, by oral gavage daily from day 6 to day 20 post-coitum inclusive, at the dose level of 100, 300 and 1000 mg/kg bw/day. One group of females was only administered with the vehicle, dried and deacidified corn oil, at the same constant dose volume of 5 mL/kg.
Individual volumes were adjusted according to the most recently recorded body weight.
- Dose selection rationale: The dose levels were selected based on the results of a limited range finder in the pregnant rat (Test Facility Study No. 20205156), on the results on the dose range finder in the non-pregnant rat (Test Facility Study No. 20164929), and in an attempt to produce graded responses to the test substance. - Maternal examinations:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Throughout the study, animals were observed for general health/mortality and moribundity twice daily, in the morning and at the end of the working day. Animals showing pain, distress or discomfort which was considered not to be transient in nature or was likely to become more severe, were sacrificed for humane reasons based on OECD guidance document on humane endpoints (ENV/JM/MONO/ 2000/7).
- Cage side observations checked in table [No.?] were included.
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Clinical observations were performed once daily from Day 2 post-coitum and lasting up to the day prior to necropsy. Females were only observed immediately after dosing.
BODY WEIGHT: Yes
- Time schedule for examinations: Animals were weighed individually on Days 2, 6, 9, 12, 15, 18 and 21 post-coitum. In order to monitor its health status, animal number 28 from the 100 mg/kg bw/day group were also weighed on Day 7, 8 and 14 post-coitum.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
Time scheule: Food consumption was quantitatively measured for Days 2-6, 6-9, 9-12, 12-15, 15-18 and 18-21 post-coitum
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes / No / No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes / No / No data
- Time schedule for examinations:
POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on day 20 post-coitum
- Organs examined: All animals (including animals sacrificed before planned necropsy) were subjected to an external, thoracic and abdominal examination, with special attention being paid to the reproductive organs. All macroscopic abnormalities were recorded, collected and fixed in 10% buffered formalin (neutral phosphate buffered 4% formaldehyde solution). - Ovaries and uterine content:
- The ovaries and uterine content was examined after termination: Yes
Each ovary and uterine horn of all surviving animals was dissected and examined as quickly as possible to determine:
- Gravid uterus weight: Yes (not for animals sacrificed before planned necropsy)
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes / No / No data
- Number of late resorptions: Yes / No / No data
- Other:
- The number and distribution of live and dead fetuses.
- The number and distribution of embryo-fetal deaths.
- The sex of each fetus based on the ano-genital distance.
In case no macroscopically visible implantation sites were present, non-gravid uteri were stained using the Salewski technique in order to detect any former implantation sites. - Fetal examinations:
- - External examinations: Yes: all viable fetuses
- Soft tissue examinations: Yes: half of all viable fetuses
- Skeletal examinations: Yes: half of all viable fetuses
- Head examinations: Yes: half of all viable fetuses - Statistics:
- Data were analysed between the control and treatment groups (low, mid and high doses) as appropriate depending on data availability
- Indices:
- An overall Fisher’s exact test was used to compare all groups at the 5% significance level. The above pairwise comparisons were conducted using a two-sided Fisher’s exact test at the 5% significance level if the overall test was significant.
No statistics were applied for data on maternal survival, pregnancy status, group mean numbers of dead fetuses, early and late resorptions, and pre- and postimplantation loss. - Historical control data:
- The laboratory has historical control data in regard to developmental/teratological parameters for this strain of rat.
- Clinical signs:
- no effects observed
- Description (incidence and severity):
- No clinical signs of toxicological relevance (such as hunched posture, flat gait, rales, piloerection, ptosis, uncoordinated movements and/or pale appearance) were noted at any exposure dose during the observation period.
- Mortality:
- no mortality observed
- Description (incidence):
- No mortalities were observed in the controls and all the treatment groups.
- Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- Mean body weights, body weight gain and weight gain corrected for gravid uterus of treated females were unaffected by treatment with the test substance up to 1000 mg/kg bw/day.
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Description (incidence and severity):
- A subjective appraisal of water consumption was maintained during the study, but no quantitative investigation was conducted as no effect was suspected.
- Ophthalmological findings:
- not specified
- Haematological findings:
- not specified
- Clinical biochemistry findings:
- not specified
- Endocrine findings:
- no effects observed
- Description (incidence and severity):
- Analysis of the samples showed that mean total T3 and total T4 levels were unaffected in animals at all the exposure doses. Mean levels of thyroid stimulating hormone (TSH) were statistically significantly increased at 1000 mg/kg/day (1.74x of controls), but there was variability in the observed responses with a coefficient of variation of 57%. Individual values remained within the historical control range of 0.129 to 0.724 (µIU/ml) for 17 of the 22 females (77%), though for the other 5/22 (23%) animals the values were above the 95%ile control limit.
- Urinalysis findings:
- not specified
- Behaviour (functional findings):
- not examined
- Immunological findings:
- not specified
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Description (incidence and severity):
- There were no test substance-related alterations in thyroid gland weights up to 1000 mg/kg/day.
- Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- No toxicologically relevant abnormalities were observed at necropsy in exposed animals up to 1000 mg/kg bw/day
- Neuropathological findings:
- not specified
- Histopathological findings: non-neoplastic:
- no effects observed
- Description (incidence and severity):
- There were no test substance-related microscopic observations in the thyroid glands up to 1000 mg/kg/day.
- Histopathological findings: neoplastic:
- no effects observed
- Description (incidence and severity):
- No neoplastic histopathological effects were evident in the tissues of animals from the control and treatment groups.
- Other effects:
- no effects observed
- Number of abortions:
- not examined
- Pre- and post-implantation loss:
- no effects observed
- Description (incidence and severity):
- There were no treatment-related effects on pre- and post-implantation losses at any of the exposure doses, compared to the controls.
- Total litter losses by resorption:
- no effects observed
- Description (incidence and severity):
- There were no treatment-related effects on total losses by resorption at any of the exposure doses, compared to the controls
- Early or late resorptions:
- no effects observed
- Description (incidence and severity):
- There were no treatment-related effects on early or late resorptions at any of the exposure doses, compared to the controls.
- Dead fetuses:
- no effects observed
- Description (incidence and severity):
- There were no treatment-related effects on the number of dead foetuses at any of the exposure doses, compared to the controls.
- Changes in pregnancy duration:
- not examined
- Changes in number of pregnant:
- no effects observed
- Description (incidence and severity):
- There were no treatment-related effects on the number of pregnant females at any of the exposure doses, compared to the controls.
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 1 000 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Basis for effect level:
- other: maternal toxicity
- Details on embryotoxic / teratogenic effects:
- No
No toxicologically significant changes were noted in any of the developmental parameters investigated in this study (i.e. litter size, post-implantation loss, fetal sex ratio, fetal body weights, fetal ano-genital distance and external, visceral and skeletal malformations and developmental variations in fetuses) after treatment up to 1000 mg/kg/day. - Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 1 000 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Basis for effect level:
- other: embrytoxicity
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 1 000 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Basis for effect level:
- other: fetotoxicity
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 1 000 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Basis for effect level:
- other: teratogenicity
- Key result
- Abnormalities:
- no effects observed
- Description (incidence and severity):
- There were no test substance-related effects on fetal external malformations and variations, visceral morphology and skeletal morphology following treatment up to 1000 mg/kg bw/day.
- Key result
- Developmental effects observed:
- no
- Conclusions:
- In the absence of adverse findings at the highest dose level, the systemic maternal No Observed Adverse Effect Level (NOAEL) for Cashew Nutshell Extract, Decarboxylated, Distillation Residue (Distillation Residue Grade) was determined to be 1000 mg/kg bw/day.
The developmental NOAEL for Distillation Residue Grade was also established as being 1000 mg/kg bw/day.
Note: In the study the mean thyroid stimulating hormone (TSH) level was increased in adult rats exposed to the test-substance at 1000 mg/kg/day/day. However, the mean TSH value of females at 1000 mg/kg/day remained within the historical control range for 77% of females. The test report concluded that “under the conditions of this study no adverse effect was observed that could be linked to the increase in TSH and therefore it was not taken into account when determining the maternal NOAEL - Executive summary:
This study was conducted to evaluate the embryotoxicity, fetotoxicity and teratogenicity of the test material in the pregnant Wistar Han rat. Test material was dissolved in dried and deacidified corn oil and administered daily by gastric intubation during the Day 6 to 20post-coitum, with the dosing solutions being prepared fresh weekly. Dose levels were 100, 300 and 1000 mg/kg bw/day which were selected based on the results of a limited range finder in the pregnant rat (Test Facility Study No. 20205156) and on the results on the dose range finder in the non-pregnant rat (Test Facility Study No. 20164929). The study included a vehicle (dried and deacidified corn oil) treated control group. Each study group contained 22 mated females.
Analysis of the samples showed that mean total T3 and T4 levels, as well as thyroid weight and histopathology, were unaffected in animals at all the exposure doses. Mean levels of thyroid stimulating hormone (TSH) were statistically significantly increased at 1000 mg/kg/day (1.74x of controls), but there was variability in the observed responses with a coefficient of variation of 57%. Individual values remained within the historical control range of 0.129 to 0.724 (µIU/ml) for 17 of the 22 females (77%), though for the other 5/22 (23%) animals the values were above the 95%ile control limit. The test report concluded that “under the conditions of this study no adverse effect was observed that could be linked to the increase in TSH and therefore it was not taken into account when determining the maternal NOAEL”.
No toxicologically significant changes at exposure levels up to 1000 mg/kg bw/day were noted in any of the remaining maternal parameters investigated in this study (i.e. mortality/moribundity, clinical appearance, body weight, food consumption, gross pathology, thyroid weights, uterine contents, histopathologic examination (thyroid gland), corpora lutea, implantation sites and pre- and pos-timplantation loss).
No test substance-related changes at exposure levels up to 1000 mg/kg bw/day were noted in any of the developmental parameters investigated in this study (i.e. litter size, sex ratio, fetal body weights, fetal anogenital distance and external, visceral and skeletal malformations and variations in fetuses).
No effects on endocrine-disruption related apical morphological and histological endpoints (i.e. anogenital distance and sex ratio in foetuses, thyroid weight and histopathology in females) and indicators of thyroid hormonal activity in females (i.e. T3, T4 and TSH levels) were observed in the OECD TG414 study at exposure doses up to 1000 mg/kg bw/day.
Reference
Effect on developmental toxicity: via oral route
- Endpoint conclusion:
- no adverse effect observed
- Dose descriptor:
- NOAEL
- 1 000 mg/kg bw/day
Effect on developmental toxicity: via inhalation route
- Endpoint conclusion:
- no study available
Effect on developmental toxicity: via dermal route
- Endpoint conclusion:
- no study available
Additional information
For the assessment of developmental toxicity to Distillation Residue Grade additional information is available for the read across substance is Cashew (Anacardium occidentale) Nutshell Extract, Decarboxylated, Distilled (Distilled Grade). A document which justifies the read–across from Cashew (Anacardium occidentale) Nutshell Extract, Decarboxylated, Distilled (Distilled Grade) to Cashew (Anacardium occidentale) Nutshell Extract, Decarboxylated, Distillation Residue (Distillation Residue Grade) is attached.
Toxicity to reproduction: other studies
Additional information
No adverse effects on reproduction or development were observed in a 28 day reproductive screening study. A similar conclusion is expected for Cashew Nutshell Extract, Decarboxylated, Distillation Residue (Distillation Residue Grade) based on the read-across justification.
A document which justifies the read–across from Cashew (Anacardium occidentale) Nutshell Extract, Decarboxylated, Distilled (Distilled Grade) to Cashew (Anacardium occidentale) Nutshell Extract, Decarboxylated, Distillation Residue (Distillation Residue Grade) is attached.
Justification for classification or non-classification
In a subchronic toxicity study (OECD TG414) Cashew Nutshell Extract, Decarboxylated, Distillation Residue (Distillation Residue Grade) was administered to Wistar Han rats at dose levels of 0, 150, 300, or 1000 mg/kg bw/day). No toxicologically significant changes at exposure levels up to 1000 mg/kg bw/day were noted in any of the remaining maternal parameters investigated in this study (i.e. mortality/moribundity, clinical appearance, body weight, food consumption, gross pathology, thyroid weights, uterine contents, histopathologic examination (thyroid gland), corpora lutea, implantation sites and pre- and pos-timplantation loss). No test substance-related changes at exposure levels up to 1000 mg/kg bw/day were noted in any of the developmental parameters investigated in this study (i.e. litter size, sex ratio, fetal body weights, fetal anogenital distance and external, visceral and skeletal malformations and variations in fetuses). Therefore classification is not considered appropriate.
No adverse effects on reproduction or development were observed in a 28 day reproductive screening study, therefore classification is not considered appropriate. A similar conclusion is expected for Cashew Nutshell Extract, Decarboxylated, Distillation Residue (Distilled Residue Grade) based on the read-across justification.
Additional information
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