Registration Dossier
Registration Dossier
Data platform availability banner - registered substances factsheets
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Diss Factsheets
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 942-754-1 | 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
Description of key information
Read-across: Combined repeated dose toxicity study with the reproduction/development toxicity screening test: NOAEL = 800 mg/kg bw/day (OECD 422, GLP, rel. 1)
Key value for chemical safety assessment
- Toxic effect type:
- dose-dependent
Repeated dose toxicity: via oral route - systemic effects
Link to relevant study records
- Endpoint:
- short-term repeated dose toxicity: oral
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- From 2008-09-29 to 2009-08-27
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Remarks:
- Study performed according to OECD test guideline No. 422 and in compliance with GLP
- Reason / purpose for cross-reference:
- reference to same study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: EPA OPPTS 870.3650 (Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test)
- Deviations:
- no
- Principles of method if other than guideline:
- not applicable
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- OECD GLP (inspected on 05th to 09th and 26th to 30th November 2007, Signed on 12th November 2008)
- Limit test:
- no
- Species:
- rat
- Strain:
- other: HanRcc: WIST(SPF)
- Details on species / strain selection:
- Recognized by international guidelines as a recommended test system.
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Harlan Laboratories Ltd. Laboratory Animal Services Wölferstrasse 44414 Füllinsdorf / Switzerland
- Age at study initiation: 11 weeks
- Weight at study initiation: Males: 284 to 318 g; Females: 178 to 214 g
- Fasting period before study: none
- Housing: Individually in Makrolon type-3 cages with wire mesh tops and sterilized standard softwood bedding (‘Lignocel’ Schill AG, 4132 Muttenz / Switzerland). During the pre-pairing period, cages with males were interspersed amongst those holding females to promote the development of regular oestrus cycles.
- Diet (e.g. ad libitum): Pelleted standard Kliba Nafag 3433 rat/mouse maintenance diet (Provimi Kliba SA, 4303 Kaiseraugst / Switzerland) was available ad libitum (batch no. 31/08).
- Water (e.g. ad libitum): Community tap-water from Füllinsdorf was available ad libitum in water bottles
- Acclimation period: 7 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): Air-conditioned with 10 - 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12-hour fluorescent light / 12-hour dark cycle with music during the light period
IN-LIFE DATES: From: 2008-09-28 To: 2009-08-27 - Route of administration:
- oral: gavage
- Vehicle:
- corn oil
- Details on oral exposure:
- PREPARATION OF DOSING SOLUTIONS:
The test material was weighed into a glass beaker on a tarred precision balance and approximately 80% of the vehicle was added (w/v). Using an appropriate homogenizer, a homogeneous suspension was prepared. Having obtained a homogeneous mixture, the remaining vehicle was added.
Separate formulations were prepared for each concentration. Homogeneity of the test item in the vehicle was maintained during the daily administration period using a magnetic stirrer.
VEHICLE
- Amount of vehicle (if gavage): 5 mL/kg bw
- Lot/batch no. (if required): 24897436 - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- On the first treatment day, one sample (middle) from the control group as well as three samples (top, middle and bottom) of about 2 g of each concentration were taken prior to dosing for analysis of concentration and homogeneity. Samples of about 2 g of each concentration were taken from the middle only to confirm stability (7 days). During the last week of the treatment, samples were taken from the middle to confirm concentration. The aliquots for analysis of dose
formulations were frozen (-20 ± 5 °C) and delivered on dry ice to Dr. D. Flade (Harlan Laboratories Ltd., Itingen / Switzerland) and stored there at -20 ± 5 °C until analysis. The samples were analysed by GC coupled to an FI detector following an analytical procedure provided by the Sponsor and adapted at Harlan Laboratories. The test item was used as the analytical standard. Analysed samples were not discarded without written consent from the study director.
The identity of the test material was confirmed by its retention time, which was similar to that measured in the working standards. The application formulations investigated during the study were found to comprise the test material in the range of 91.7% to 108.9% and thus, the required content limit of ±20% with reference to the nominal concentration was met. The homogeneous distribution of the test material in the preparations was approved because single results did not
deviate more than 5.9% (<15%) from the corresponding mean. The application formulations were considered to be stable for at least 7 days when kept at room temperature. - Duration of treatment / exposure:
- Males: minimum 4 weeks. Females: approximately 7 weeks.
- Frequency of treatment:
- Daily
- Dose / conc.:
- 100 mg/kg bw/day (actual dose received)
- Dose / conc.:
- 400 mg/kg bw/day (actual dose received)
- Dose / conc.:
- 800 mg/kg bw/day (actual dose received)
- No. of animals per sex per dose:
- 10 animals/sex/dose
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: The dose levels were selected based on a previous dose range finding toxicity study in Han Wistar Rats, Harlan Laboratories Study C05595, using dose levels of 100, 300, and 1000 mg/kg bw/day.
- Rationale for animal assignment (if not random): Computer-generated random algorithm. In addition body weights (recorded on the day of allocation) were taken into consideration in order to ensure similar mean body weights in all groups. - Positive control:
- none
- Observations and examinations performed and frequency:
- VIABILITY / MORTALITYS: Yes
- Time schedule: Twice daily
CLINICAL SIGNS: Yes
- Time schedule: Daily cage-side clinical observations (once daily during acclimatization and up to day of necropsy).
Additionally females were observed for signs of difficult or prolonged parturition, and behavioural abnormalities in nesting and nursing.
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Once prior to the first administration of the test item and weekly thereafter, performed outside the home cage. Animals were observed
- Parameters examined: changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions, and autonomic activity (e.g. lacrimation, piloerection, pupil size, unusual respiratory pattern). Changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies or bizarre behaviour were also reported.
BODY WEIGHT: Yes
- Time schedule for examinations: Recorded daily from treatment start to day of necropsy.
FOOD CONSUMPTION:
Males: Weekly during pre-pairing and after pairing periods
Females: Pre-pairing period days 1-8, 8-14 and 14-16; gestation days 0-7, 7-14 and 14-21 post coitum, and days 1-4 post partum.
No food consumption was recorded during the pairing period.
OPHTHALMOSCOPIC EXAMINATION: No
HAEMATOLOGY: Yes
- Time schedule for collection of blood: Blood samples were obtained on the day before or on the day of the scheduled necropsy from
5 males from each group. Blood samples from 5 lactating females from each group were obtained on day 5 post partum
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes, approximately 18 hours before blood sampling but allowed access to water ad libitum
- How many animals: 5 animals/sex/dose
- Parameters checked in table 5.7.1/2.
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Blood samples were obtained on the day before or on the day of the scheduled necropsy from
5 males from each group. Blood samples from 5 lactating females from each group were obtained on day 5 post partum
- Animals fasted: Yes, approximately 18 hours before blood sampling but allowed access to water ad libitum
- How many animals:5 animals/sex/dose
- Parameters checked in table 5.7.1/2 were examined.
URINALYSIS: No
NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: males shortly before the scheduled sacrifice and females on day 3 or 4 post partum
- Dose groups that were examined: five P generation males and five P generation females from each group (erroneously six females were investigated in group 4)
- Battery of functions tested:
a) Cage-side observations: unusual body movements (e.g. tremors, convulsions), abnormal behaviour (e.g. circling, stereotypy) and posture as well as resistance to removal.
b) Hand-held observations: palpebral closure, pinna reflex, lacrimation, pupil size, pupil reactivity, salivation, muscle tone, extensor thrust response, righting reflex and reaction to handling.
c) Open field observations: level of ambulatory activity including rearing (one minute evaluation), responsiveness to sharp noise, paw pinch, gait evaluation, quantity of urine and fecal pellets voided.
d) Categorical observations (can be made any time during the FOB): hair coat, behavior, respiration, muscle movements, eyes, hearing ability (Preyer’s reflex), urine or feces, soiling, general abnormalities, posture.
e) Measurements / Counts: hind limb / fore limb grip strength, landing foot splay, rectal temperature.
Additionally, locomotor activity was measured quantitatively for the same animals. Activity was measured with an Activity Monitor AMS-0151 (FMI, Germany). Activity of the animals (based on beam count) was recorded for 6-minute intervals over a period of 30 minutes. - Sacrifice and pathology:
- Males were sacrificed after treatment of at least 28 days, when no longer needed for the assessment of reproductive effects. Pups were sacrificed on day 4 post partum. Dams were sacrificed on day 5 post partum. If birth did not occur on the expected date (day 21 post coitum), the dam was sacrificed on day 25 post coitum.
GROSS PATHOLOGY: Yes (see table 7.5.1/3)
HISTOPATHOLOGY: Yes (see table 7.5.1/3) - Other examinations:
- The testes were stained by PAS hematoxylin for qualitative sperm staging.
- Statistics:
- The following statistical methods were used to analyse food consumption, body weights and reproduction data:
- Means and standard deviations of various data were calculated.
- The Dunnett-test (many to one t-test) based on a pooled variance estimate was applied if the variables could be assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
- The Steel-test (many-one rank test) was applied instead of the Dunnett test when the data could not be assumed to follow a normal distribution.
- Fisher's exact-test was applied to the macroscopical findings. - Clinical signs:
- effects observed, non-treatment-related
- Description (incidence and severity):
- All males and females treated with 400 or 800 mg/kg bw/d pushed their head through the bedding after application from day 14 of the pre-pairing period onwards. This was considered to be a sign of discomfort following treatment rather than a toxic effect of the test item. One male and one female treated with 800 mg/kg bw/d had salivation for isolated days. In addition, one female had salivation for most of the pre-pairing, pairing and gestation periods. This was considered to be a sign of discomfort following the treatment.
Other clinical signs noted were a wound on the shoulder of one male in low-dose group and hair loss in one dam in the control group. These findings were considered to be incidental. - Mortality:
- no mortality observed
- Body weight and weight changes:
- effects observed, non-treatment-related
- Description (incidence and severity):
- - Males: In high-dose group, body weight gain was statistically significantly reduced on days 3, 6 and 10 of the pre-pairing period and on the last day of the after pairing period. However, mean body weight was similar in all dose groups throughout the study. No test item-related effects were noted in the other groups.
- Females: In high-dose group 4, mean body weight gain was statistically significantly reduced over days 2 - 7 of the pre-pairing period and absolute body weight on days 3, 4 and 6 during the pre-pairing period. This was considered to be a result of treatment with the test item. No other test item-related effects were noted during the study at any dose level. - Food consumption and compound intake (if feeding study):
- effects observed, non-treatment-related
- Description (incidence and severity):
- Normal responses to the taste of fragrances
- Males: Mean food consumption was very slightly reduced in medium- and high-dose groups during the first week of the pre-pairing period (-5.6% and -6.1%, respectively). Thereafter, food consumption was not affected by treatment with the test item in any dose group.
- Females: In high-dose group, mean food consumption was statistically significantly reduced over the first week of the pre-pairing period (-18.4% compared to the control group). This reduction was considered to be a test item-related effect. The statistically significant increase in low-dose group in the first week of the pre-pairing period was considered to be incidental due to the lack of a dose-dependent pattern.
No other test item-related effects were noted in the food consumption during the study at any dose level. - Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- not examined
- Haematological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Statistically significant increased platelets counts in medium- and high-dose males; prothrombin time outside the historical control data in high-dose females. Not adverse
- Males The level of platelets was statistically significantly increased in medium-dose groups (+23.2%, compared to the control group) and 4 (+33.8%) and was outside the historical control data in high-dose group.
- Females: In the females in high-dose group, although the prothrombin time was not statistically significantly increased (+15.3%), the level was outside the historical control data.
All other values noted for the males and females, including statistically significant changes, were within the range of the historical control data. - Clinical biochemistry findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Increased cholesterol and globulin level in medium- and high-dose animals; increased globulin levels in medium- and high-dose males. Not adverse.
- Males The level of cholesterol was statistically significantly increased in medium- and high-dose dose groups (+28.5% and +44.0%, respectively). Both these values were outside the range of the historical control data. The level of protein (+5.3% in medium-dose group and +8.6% in high-dose group) as well as globulin (+9.6% in medium-dose group and +16.5% in high-dose group) were increased and although the values were not statistically significant, they were outside the range of the historical control data.
- Females: The level of cholesterol in medium- and high-dose groups (+59.0% and +125.5%, respectively) was outside the range of the historical control data and was statistically significantly increased in high-dose group. The level of globulin was statistically significantly increased in medium- and high-dose groups (+17.3% and +16.1%, respectively) and both levels were outside the range of the historical control data.
All other values noted for the males and females, including statistically significant changes, were within the range of the historical control data. - Urinalysis findings:
- not examined
- Behaviour (functional findings):
- effects observed, treatment-related
- Description (incidence and severity):
- Reduced number of rearing in high-dose males. Dose-dependent decrease in body temperature.
In high-dose group, there was an increased incidence in males of a reduced number of rearings (80%) compared to the control group (20%). In addition, body temperature was statistically significantly decreased in males (37.9 °C compared to the control group 38.5 °C) and females (38.3 °C compared to 39.0 °C in the control group).
In medium-dose group, the body temperature of the females was statistically significantly reduced (38.5 °C compared to 39.0 °C in the control group).
Although the body temperatures were within the range of the historical control data, since they decreased in a dose-dependent manner, it may be a slight effect of the test item.
No other test item-related effects were noted. - Immunological findings:
- not examined
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Description (incidence and severity):
- The weights and ratios of the liver were statistically significantly increased in males and females in the groups receiving the test item. In medium- and high-dose groups, this corresponded to histopathological findings. In low-dose group, since no test item-related histopathological findings were noted and the mean absolute weight was within the historical control data, the increase was considered to be incidental.
The weights and organ/body weight ratios of the kidney were statistically significantly increased in high-dose group in the females. Since this did not correspond to any histopathological findings and the mean absolute weight was within the historical control data, it was therefore considered to be incidental. - Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- 2 females in high-dose group had an enlarged liver. No other test item-related findings were noted for males or females in any group.
- Neuropathological findings:
- not examined
- Histopathological findings: non-neoplastic:
- no effects observed
- Description (incidence and severity):
- In the liver, there was centrilobular hepatocellular hypertrophy in males and females in medium- and high-dose groups. There was no further lesion along with this hypertrophy and therefore, an adaptive change in liver metabolism was concluded.
In the thyroid glands, there was diffuse follicular hypertrophy in some high dose males and females at minor degrees of severity. This was considered to be the result of increased metabolism in the liver. The changes in the liver and in the thyroid glands were not considered to be adverse.
In the kidneys of males in medium- and high-dose groups, there were increased degrees of severity of hyaline inclusions along with an increased incidence and severity of tubular basophilia. It is considered that the hyaline inclusions represent alpha-2-microglobulin and the increased deposition is a consequence of increased liver metabolism. The nature of this lesion may be considered to be adverse in male rats due to the increased tubular basophilia that represent regeneration from a previous tubulopathy.
In low-dose group, no test item-related findings were noted. - Histopathological findings: neoplastic:
- no effects observed
- Other effects:
- no effects observed
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 800 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: The induced hyaline droplet nephropathy in male rats is known to be a rat-specific lesion and is not relevant for human
- Key result
- Critical effects observed:
- no
- Conclusions:
- Based on the results of the study and since the induced hyaline droplet nephropathy in male rats is known to be a rat-specific lesion and not relevant for human, the general NOAEL was considered to be 800 mg/kg bw/day.
- Executive summary:
In a combined repeated dose toxicity study with the reproduction/development toxicity screening test performed in accordance with OECD test guideline No. 422 and in compliance with GLP, the test substance diluted in corn oil was administered to 10 HanRcc: WIST(SPF) rats/sex/dose by gavage at dose levels of 0, 100, 400 and 800 mg/kg bw/day. Control rats were given the vehicle alone. The test substance was administered to male rats for at least 28 days and to female rats for 16 days prior to pairing, through the pairing and gestation periods until the F1 generation reached day 4 post partum.
The following results were obtained:
- Mortality and General tolerability
All males and females in medium- and high dose groups pushed their head through the bedding after application from day 14 of the pre-pairing period onwards. One male and one female in high-dose group had salivation for isolated days. In addition, one female had salivation for most of the pre-pairing, pairing and gestation periods. These findings were considered to be a sign of discomfort following the treatment.
- Functional Observational Battery
Body temperature was statistically significantly decreased in high-dose group in males and females as well as in the females in medium-dose group. Body temperature decreased in a dose-dependent manner. However, since these changes were within the range of the historical control data, they were considered of no toxicological importance.
There was an increased incidence of a reduced number of rearings in high-dose group compared to the control group. However, in general, when only one FOB measurement is affected, the results are not considered evidence of a neurotoxic effect, mostly when the effects occurred at the high dose, in the presence of sign of toxicity such as body weigh or body temperature descreases (Guidelines for Neurotoxicity Risk Assessment, EPA, April 1998).
- Food consumption
In the males, mean food consumption was very slightly reduced in medium- and high-dose groups during the first week of the pre-pairing period. In the females, it was statistically significantly reduced in high-dose group over the first week of the pre-pairing period. These variations are normal responses to the taste of fragrances, like ST 10 C 08.
- Body weights
In high-dose group in the males, body weight gain was statistically significantly reduced on isolated days during the pre-pairing period and on the last day of the after pairing period. In the females, mean body weight gain was statistically significantly reduced over days 2 - 7 of the pre-pairing period and absolute body weight was occasionally statistically significantly reduced. These findings were correlated to lower mean food consumption and were fully reversible; therefore they are not considered as toxicologically significant.
- Clinical Laboratory Investigations
Haematology: The level of platelets was statistically significantly increased in the males in medium- and high-dose groups (+23.2 and +33.8%, respectively) and was outside the historical control data in high-dose group. In the females in high-dose group, although the prothrombin time was not statistically significantly increased, the level was outside the range of the historical control data.
Clinical biochemistry: In both males and females, the levels of cholesterol and globulin in medium- and high-dose groups were outside the range of the historical control data. The level of protein in the males in the same groups was outside the range of the historical control data.
These changes were considered as adaptive in nature, and of no toxicological concern.
- Organ Weights
The weights and ratios of the liver were statistically significantly increased in males and females in medium- and high-dose groups.
- Macroscopical Findings and Histopathological Examinations
At macroscopical examination, two females in high-dose group were noted to have an enlarged liver.
In this dose group, in the liver there was centrilobular hepatocellular hypertrophy and, as a result, diffuse follicular hypertrophy in the thyroid glands in males and females at minor degrees of severity.
These findings were considered to be adaptive responses to the test material and not to be adverse. In the kidneys of males, there were increased degrees of severity of hyaline inclusions along with an increased incidence and severity of tubular basophilia. It is considered that the hyaline inclusions representα2-microglobulin. It may be considered that the increased deposition is a consequence of increased liver metabolism. The nature of this lesion may be considered to be adverse in rat males due to the increased tubular basophilia that represent regeneration from a previous tubulopathy.
In medium-dose group, there was centrilobular hepatocellular hypertrophy in males and females. In addition, in the kidneys of the males, there were increased degrees of severity of hyaline inclusions along with an increased incidence and severity of tubular basophilia.
Based on the results of the study and since the induced hyaline droplet nephropathy in male rats is known to be a rat-specific lesion and not relevant for human, the general NOAEL was considered to be 800 mg/kg bw/day.
Based on the results of this study, the test substance is not classified for damage to organs through prolonged oral repeated exposure according to the criteria of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.
This study is considered as acceptable and satisfies the requirement for sub-acute oral toxicity endpoint.
- Endpoint:
- short-term repeated dose toxicity: oral
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
[Further information is included as attachment to Iuclid section 13]
1. HYPOTHESIS FOR THE ANALOGUE APPROACH
This read-across approach is based on the hypothesis that the source and target substances have similar physico-chemical, toxicological, ecotoxicological and environmental fate properties because of their structural similarity.
2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
The target and the source substances (Table 1) are Cycloalkane ethers. The source substance is the racemic form of the 9bR isomer, while the target substance is a reaction mass of the source together with the 9bS isomer.
3. ANALOGUE APPROACH JUSTIFICATION
The source and the target substances are expected to have the same toxicokinetic profile (ADME) based on structural similarity (Section 3.2) and similar physico-chemical properties (Section 4.1). In addition, the available systemic and local toxicity studies (acute oral toxicity, mutagenicity, skin/eye irritation, skin sensitisation – section 4.4) are comparable between the two substances.
The source substance is therefore expected to have similar toxicological profile than the target substance.
The study design is adequate and reliable for the purpose of the prediction based on read-across. The test material used represents the source substance as described in the hypothesis in terms of purity and impurities. The results of the studies are adequate for the purpose of classification and labelling.
Therefore, based on the considerations above, it can be concluded that the results of the sub-acute toxicity study conducted in the rat with the source substance are likely to predict the properties of the target substance and are considered as adequate to fulfil the information requirement of Annex VIII, 8.6.1.
4. DATA MATRIX
See Iuclid section 13 - Reason / purpose for cross-reference:
- read-across source
- Reason / purpose for cross-reference:
- read-across: supporting information
- Clinical signs:
- effects observed, non-treatment-related
- Description (incidence and severity):
- All males and females treated with 400 or 800 mg/kg bw/d pushed their head through the bedding after application from day 14 of the pre-pairing period onwards. This was considered to be a sign of discomfort following treatment rather than a toxic effect of the test item. One male and one female treated with 800 mg/kg bw/d had salivation for isolated days. In addition, one female had salivation for most of the pre-pairing, pairing and gestation periods. This was considered to be a sign of discomfort following the treatment.
Other clinical signs noted were a wound on the shoulder of one male in low-dose group and hair loss in one dam in the control group. These findings were considered to be incidental. - Mortality:
- no mortality observed
- Body weight and weight changes:
- effects observed, non-treatment-related
- Description (incidence and severity):
- - Males: In high-dose group, body weight gain was statistically significantly reduced on days 3, 6 and 10 of the pre-pairing period and on the last day of the after pairing period. However, mean body weight was similar in all dose groups throughout the study. No test item-related effects were noted in the other groups.
- Females: In high-dose group 4, mean body weight gain was statistically significantly reduced over days 2 - 7 of the pre-pairing period and absolute body weight on days 3, 4 and 6 during the pre-pairing period. This was considered to be a result of treatment with the test item. No other test item-related effects were noted during the study at any dose level. - Food consumption and compound intake (if feeding study):
- effects observed, non-treatment-related
- Description (incidence and severity):
- Normal responses to the taste of fragrances
- Males: Mean food consumption was very slightly reduced in medium- and high-dose groups during the first week of the pre-pairing period (-5.6% and -6.1%, respectively). Thereafter, food consumption was not affected by treatment with the test item in any dose group.
- Females: In high-dose group, mean food consumption was statistically significantly reduced over the first week of the pre-pairing period (-18.4% compared to the control group). This reduction was considered to be a test item-related effect. The statistically significant increase in low-dose group in the first week of the pre-pairing period was considered to be incidental due to the lack of a dose-dependent pattern.
No other test item-related effects were noted in the food consumption during the study at any dose level. - Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- not examined
- Haematological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Statistically significant increased platelets counts in medium- and high-dose males; prothrombin time outside the historical control data in high-dose females. Not adverse
- Males The level of platelets was statistically significantly increased in medium-dose groups (+23.2%, compared to the control group) and 4 (+33.8%) and was outside the historical control data in high-dose group.
- Females: In the females in high-dose group, although the prothrombin time was not statistically significantly increased (+15.3%), the level was outside the historical control data.
All other values noted for the males and females, including statistically significant changes, were within the range of the historical control data. - Clinical biochemistry findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Increased cholesterol and globulin level in medium- and high-dose animals; increased globulin levels in medium- and high-dose males. Not adverse.
- Males The level of cholesterol was statistically significantly increased in medium- and high-dose dose groups (+28.5% and +44.0%, respectively). Both these values were outside the range of the historical control data. The level of protein (+5.3% in medium-dose group and +8.6% in high-dose group) as well as globulin (+9.6% in medium-dose group and +16.5% in high-dose group) were increased and although the values were not statistically significant, they were outside the range of the historical control data.
- Females: The level of cholesterol in medium- and high-dose groups (+59.0% and +125.5%, respectively) was outside the range of the historical control data and was statistically significantly increased in high-dose group. The level of globulin was statistically significantly increased in medium- and high-dose groups (+17.3% and +16.1%, respectively) and both levels were outside the range of the historical control data.
All other values noted for the males and females, including statistically significant changes, were within the range of the historical control data. - Urinalysis findings:
- not examined
- Behaviour (functional findings):
- effects observed, treatment-related
- Description (incidence and severity):
- Reduced number of rearing in high-dose males. Dose-dependent decrease in body temperature.
In high-dose group, there was an increased incidence in males of a reduced number of rearings (80%) compared to the control group (20%). In addition, body temperature was statistically significantly decreased in males (37.9 °C compared to the control group 38.5 °C) and females (38.3 °C compared to 39.0 °C in the control group).
In medium-dose group, the body temperature of the females was statistically significantly reduced (38.5 °C compared to 39.0 °C in the control group).
Although the body temperatures were within the range of the historical control data, since they decreased in a dose-dependent manner, it may be a slight effect of the test item.
No other test item-related effects were noted. - Immunological findings:
- not examined
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Description (incidence and severity):
- The weights and ratios of the liver were statistically significantly increased in males and females in the groups receiving the test item. In medium- and high-dose groups, this corresponded to histopathological findings. In low-dose group, since no test item-related histopathological findings were noted and the mean absolute weight was within the historical control data, the increase was considered to be incidental.
The weights and organ/body weight ratios of the kidney were statistically significantly increased in high-dose group in the females. Since this did not correspond to any histopathological findings and the mean absolute weight was within the historical control data, it was therefore considered to be incidental. - Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- 2 females in high-dose group had an enlarged liver. No other test item-related findings were noted for males or females in any group.
- Neuropathological findings:
- not examined
- Histopathological findings: non-neoplastic:
- no effects observed
- Description (incidence and severity):
- In the liver, there was centrilobular hepatocellular hypertrophy in males and females in medium- and high-dose groups. There was no further lesion along with this hypertrophy and therefore, an adaptive change in liver metabolism was concluded.
In the thyroid glands, there was diffuse follicular hypertrophy in some high dose males and females at minor degrees of severity. This was considered to be the result of increased metabolism in the liver. The changes in the liver and in the thyroid glands were not considered to be adverse.
In the kidneys of males in medium- and high-dose groups, there were increased degrees of severity of hyaline inclusions along with an increased incidence and severity of tubular basophilia. It is considered that the hyaline inclusions represent alpha-2-microglobulin and the increased deposition is a consequence of increased liver metabolism. The nature of this lesion may be considered to be adverse in male rats due to the increased tubular basophilia that represent regeneration from a previous tubulopathy.
In low-dose group, no test item-related findings were noted. - Histopathological findings: neoplastic:
- no effects observed
- Other effects:
- no effects observed
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 800 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: The induced hyaline droplet nephropathy in male rats is known to be a rat-specific lesion and is not relevant for human
- Key result
- Critical effects observed:
- no
- Conclusions:
- Based on the results of the study and since the induced hyaline droplet nephropathy in male rats is known to be a rat-specific lesion and not relevant for human, the general NOAEL was considered to be 800 mg/kg bw/day for the source subtance. The target substance has the same molecular weight than the source substance so no correction is needed for this NOAEL which is therefore set to 800 mg/kg bw/day for the target substance.
- Executive summary:
In a combined repeated dose toxicity study with the reproduction/development toxicity screening test performed in accordance with OECD test guideline No. 422 and in compliance with GLP, the source substance diluted in corn oil was administered to 10 HanRcc: WIST(SPF) rats/sex/dose by gavage at dose levels of 0, 100, 400 and 800 mg/kg bw/day. Control rats were given the vehicle alone. The source substance was administered to male rats for at least 28 days and to female rats for 16 days prior to pairing, through the pairing and gestation periods until the F1 generation reached day 4 post partum.
The following results were obtained:
- Mortality and General tolerability
All males and females in medium- and high dose groups pushed their head through the bedding after application from day 14 of the pre-pairing period onwards. One male and one female in high-dose group had salivation for isolated days. In addition, one female had salivation for most of the pre-pairing, pairing and gestation periods. These findings were considered to be a sign of discomfort following the treatment.
- Functional Observational Battery
Body temperature was statistically significantly decreased in high-dose group in males and females as well as in the females in medium-dose group. Body temperature decreased in a dose-dependent manner. However, since these changes were within the range of the historical control data, they were considered of no toxicological importance.
There was an increased incidence of a reduced number of rearings in high-dose group compared to the control group. However, in general, when only one FOB measurement is affected, the results are not considered evidence of a neurotoxic effect, mostly when the effects occurred at the high dose, in the presence of sign of toxicity such as body weigh or body temperature descreases (Guidelines for Neurotoxicity Risk Assessment, EPA, April 1998).
- Food consumption
In the males, mean food consumption was very slightly reduced in medium- and high-dose groups during the first week of the pre-pairing period. In the females, it was statistically significantly reduced in high-dose group over the first week of the pre-pairing period. These variations are normal responses to the taste of fragrances, like ST 10 C 08.
- Body weights
In high-dose group in the males, body weight gain was statistically significantly reduced on isolated days during the pre-pairing period and on the last day of the after pairing period. In the females, mean body weight gain was statistically significantly reduced over days 2 - 7 of the pre-pairing period and absolute body weight was occasionally statistically significantly reduced. These findings were correlated to lower mean food consumption and were fully reversible; therefore they are not considered as toxicologically significant.
- Clinical Laboratory Investigations
Haematology: The level of platelets was statistically significantly increased in the males in medium- and high-dose groups (+23.2 and +33.8%, respectively) and was outside the historical control data in high-dose group. In the females in high-dose group, although the prothrombin time was not statistically significantly increased, the level was outside the range of the historical control data.
Clinical biochemistry: In both males and females, the levels of cholesterol and globulin in medium- and high-dose groups were outside the range of the historical control data. The level of protein in the males in the same groups was outside the range of the historical control data.
These changes were considered as adaptive in nature, and of no toxicological concern.
- Organ Weights
The weights and ratios of the liver were statistically significantly increased in males and females in medium- and high-dose groups.
- Macroscopical Findings and Histopathological Examinations
At macroscopical examination, two females in high-dose group were noted to have an enlarged liver.
In this dose group, in the liver there was centrilobular hepatocellular hypertrophy and, as a result, diffuse follicular hypertrophy in the thyroid glands in males and females at minor degrees of severity.
These findings were considered to be adaptive responses to the test material and not to be adverse. In the kidneys of males, there were increased degrees of severity of hyaline inclusions along with an increased incidence and severity of tubular basophilia. It is considered that the hyaline inclusions representα2-microglobulin. It may be considered that the increased deposition is a consequence of increased liver metabolism. The nature of this lesion may be considered to be adverse in rat males due to the increased tubular basophilia that represent regeneration from a previous tubulopathy.
In medium-dose group, there was centrilobular hepatocellular hypertrophy in males and females. In addition, in the kidneys of the males, there were increased degrees of severity of hyaline inclusions along with an increased incidence and severity of tubular basophilia.
Based on the results of the study and since the induced hyaline droplet nephropathy in male rats is known to be a rat-specific lesion and not relevant for human, the general NOAEL was considered to be 800 mg/kg bw/day for the source substance. The target substance has the same molecular weight than the source substance so no correction is needed for this NOAEL which is therefore set to 800 mg/kg bw/day for the target substance.
Based on the results of this study, the source and the target substances are not classified for damage to organs through prolonged oral repeated exposure according to Regulation (EC) No. 1272/2008 (CLP) and to the GHS.
This study is considered as acceptable and satisfies the requirement for sub-acute oral toxicity endpoint.
Referenceopen allclose all
Discussion:
At 800 mg/kg bw/day, salivation was noted in isolated individuals. There was an increased incidence of a decreased number of rearings in the males in group 4 compared to the control group. In addition, body temperature was statistically significantly decreased in males and females in group 4. Although this was within the range of the historical control data, since body temperature decreased in a dose-dependent manner, it may be a slight effect of the test item. Mean food consumption and body weight gain were reduced slightly and transiently in the males and females in the first week of the pre-pairing period. Clinical laboratory investigations revealed that the level of platelets was statistically significantly increased in the males. In the females, the prothrombin time was outside the range of the historical control data. In both males and females, the levels of cholesterol and globulin were outside the range of the historical control data. The level of protein in the males was also outside the range of the historical control data. At necropsy, the weights and ratios of the liver were statistically significantly increased in males and females. From the macroscopical examination, two females had an enlarged liver. Microscopically, in the liver there was centrilobular hepatocellular hypertrophy and, as a result, diffuse follicular hypertrophy in the thyroid glands in males and females at minor degrees of severity. These findings were not considered to be adverse. In the kidneys of males, there was an increased degree of severity of hyaline inclusions along with an increased incidence and severity of tubular basophilia. It is considered that the hyaline inclusions represent α2-microglobulin and that the increased deposition is a consequence of increased liver metabolism. The nature of this lesion may be considered to be adverse in rat males due to the increased tubular basophilia that represent regeneration from a previous tubulopathy.
At 400 mg/kg bw/day, mean food consumption was very slightly reduced in the males during the first week of the pre-pairing period. This had no effect on the mean absolute body weights. The level of platelets was statistically significantly increased in the males. In both males and females, the levels of cholesterol and globulin were outside the range of the historical control data. The level of protein in the males was also outside the range of the historical control data. At necropsy, the weights and ratios of the liver were statistically significantly increased in males and females. Microscopical examination revealed centrilobular hepatocellular hypertrophy in males and females. In addition, in the kidneys of the males, there were increased degrees of severity of hyaline inclusions along with an increased incidence and severity of tubular basophilia. The changes that were observed for platelets may be related to changed renal function and increased excretion. Biochemical data reveal also an affection of the female kidneys, by changes in protein and ion parameters. The cause of increased globulin levels remains unclear. Other biochemical changes recorded, including cholesterol and bilirubin are likely due to changes in the liver metabolism.
At 100 mg/kg bw/d, no test item-related effects were noted in the in-life phase. No effects were noted in the clinical laboratory investigations. At necropsy, no test item-related macroscopical or microscopical findings were noted.
Discussion:
At 800 mg/kg bw/day, salivation was noted in isolated individuals. There was an increased incidence of a decreased number of rearings in the males in group 4 compared to the control group. In addition, body temperature was statistically significantly decreased in males and females in group 4. Although this was within the range of the historical control data, since body temperature decreased in a dose-dependent manner, it may be a slight effect of the test item. Mean food consumption and body weight gain were reduced slightly and transiently in the males and females in the first week of the pre-pairing period. Clinical laboratory investigations revealed that the level of platelets was statistically significantly increased in the males. In the females, the prothrombin time was outside the range of the historical control data. In both males and females, the levels of cholesterol and globulin were outside the range of the historical control data. The level of protein in the males was also outside the range of the historical control data. At necropsy, the weights and ratios of the liver were statistically significantly increased in males and females. From the macroscopical examination, two females had an enlarged liver. Microscopically, in the liver there was centrilobular hepatocellular hypertrophy and, as a result, diffuse follicular hypertrophy in the thyroid glands in males and females at minor degrees of severity. These findings were not considered to be adverse. In the kidneys of males, there was an increased degree of severity of hyaline inclusions along with an increased incidence and severity of tubular basophilia. It is considered that the hyaline inclusions represent α2-microglobulin and that the increased deposition is a consequence of increased liver metabolism. The nature of this lesion may be considered to be adverse in rat males due to the increased tubular basophilia that represent regeneration from a previous tubulopathy.
At 400 mg/kg bw/day, mean food consumption was very slightly reduced in the males during the first week of the pre-pairing period. This had no effect on the mean absolute body weights. The level of platelets was statistically significantly increased in the males. In both males and females, the levels of cholesterol and globulin were outside the range of the historical control data. The level of protein in the males was also outside the range of the historical control data. At necropsy, the weights and ratios of the liver were statistically significantly increased in males and females. Microscopical examination revealed centrilobular hepatocellular hypertrophy in males and females. In addition, in the kidneys of the males, there were increased degrees of severity of hyaline inclusions along with an increased incidence and severity of tubular basophilia. The changes that were observed for platelets may be related to changed renal function and increased excretion. Biochemical data reveal also an affection of the female kidneys, by changes in protein and ion parameters. The cause of increased globulin levels remains unclear. Other biochemical changes recorded, including cholesterol and bilirubin are likely due to changes in the liver metabolism.
At 100 mg/kg bw/d, no test item-related effects were noted in the in-life phase. No effects were noted in the clinical laboratory investigations. At necropsy, no test item-related macroscopical or microscopical findings were noted.
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed
- Dose descriptor:
- NOAEL
- 800 mg/kg bw/day
- Study duration:
- subacute
- Species:
- rat
- Quality of whole database:
- The key study is GLP-compliant and of high quality (Klimisch score = 1)
Repeated dose toxicity: inhalation - systemic effects
Endpoint conclusion
- Endpoint conclusion:
- no study available
Repeated dose toxicity: inhalation - local effects
Endpoint conclusion
- Endpoint conclusion:
- no study available
Repeated dose toxicity: dermal - systemic effects
Endpoint conclusion
- Endpoint conclusion:
- no study available
Repeated dose toxicity: dermal - local effects
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
No study was available on the substance itself, therefore a read-across approach was used. The source substance is considered adequate for read-across purpose as data relates to the racemic form of the target substance (see Iuclid section 13 for additional justification).
In the dose range-finding study for the Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test (Harlan, 2009, rel.2), 3 rats/sex/dose were treated by gavage at the dose levels of 0, 100, 300 or 1000 mg source substance/kg bw/day. Males were treated over a 14-day pre-pairing period and during the pairing period and up to one day before necropsy (for a total of at least 28 days of treatment). Females were treated throughout the pre-pairing, pairing and gestation period up to day 13 post coitum. At the high dose level, reduced food consumption and body weight gain were noted. Due to pre-implantation loss in one of two females at 1000 mg/kg bw/day (see §7.8.1), the high-dose level to be used in the main study was lowered to 800 mg/kg bw/day. Due to the small number of females used in this range-finding study, it is difficult to estimate if this observation is related to treatment or due to an abnormal female. Using a worst-case approach, the high-dose level to be used in the main study was lowered to 800 mg/kg bw/day.
In the main study, considered as the key study (Harlan, 2009, rel. 1), the source substance diluted in corn oil was administered orally, by gavage at dose levels of 0, 100, 400 or 800 mg/kg bw/day, to rats according to the OECD test guideline No. 422 and in compliance with GLP.
No death occurred after the administration of the source substance. Some effects of discomfort, such as salivation, were observed following treatment at the two highest dose levels. Increased liver weight, biochemical changes (bilirubin, cholesterol, protein) and liver hypertrophy were considered to be part of an adaptive response to an increase in metabolic demand. The induced hyaline droplet nephropathy observed in male rats at 800 and 400 mg/kg bw/day is known to be a rat-specific lesion and therefore is not relevant for human.
The No Observed Adverse Effect Level (NOAEL) for repeated dose toxicity is therefore 800 mg/kg bw/day. The target substance has the same molecular weight than the source substance so no correction is needed for this NOAEL which is therefore set to 800 mg/kg bw/day for the target substance.
Justification for classification or non-classification
Harmonized classification:
The substance has no harmonized classification according to the Regulation (EC) No 1272/2008 (CLP).
Self-classification:
Based on the available data on a supporting substance, no additional classification is proposed regarding the specific target organ toxicity after oral dose-repeated exposure acording to CLP and to the GHS.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.