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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 936-831-9 | 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
Repeated dose toxicity: oral
Administrative data
- Endpoint:
- sub-chronic toxicity: oral
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 022
- Report date:
- 2022
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- other: OECD 421 (Reproduction/Developmental Toxicity Screening Test)
- Version / remarks:
- 29 July 2016
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
Test material
- Reference substance name:
- (9-acetoxy-3,8,10-triethyl-7,8,10-trimethyl-1,5-dioxa-9-azaspiro[5.5]undec-3-yl)methyl palmitate
- Molecular formula:
- C36 H67 N O6
- IUPAC Name:
- (9-acetoxy-3,8,10-triethyl-7,8,10-trimethyl-1,5-dioxa-9-azaspiro[5.5]undec-3-yl)methyl palmitate
Constituent 1
Test animals
- Species:
- rat
- Strain:
- Wistar
- Details on species / strain selection:
- The Wistar Hannover rat is the species and strain of choice because it is accepted by many regulatory authorities and there is ample experience and background data on this species and strain.
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Envigo RMS srl, San Pietro al Natisone (UD), Italy.
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 9 to 10 weeks old
- Weight at study initiation: 200 to 225 g for males and 175 to 200 g for females
- Housing:
The animals were housed in a limited access rodent facility. From arrival to pairing, animals were housed up to 5 of one sex to a cage, in polysulfone solid bottomed cages measuring 59.5×38×20 cm (Tecniplast Gazzada S.a.r.l., Buguggiate,Varese). Nesting material was provided inside suitable bedding bags and changed at least twice a week.
During mating, animals were housed one male to one female in clear polysulfone cages measuring 42.5×26.6×18.5 cm with a stainless steel mesh lid and floor (Tecniplast Gazzada S.a.r.l., Buguggiate, Varese). Each cage tray held absorbent material which was inspected and changed daily.
After mating, the males were re-caged as they were before mating.
The females were transferred to individual solid bottomed cages for the gestation period, birth and lactation (measuring 42.5×26.6×18.5 cm). Nesting material was provided inside suitable bedding bags. In addition, suitable nesting material (Scobis 0 Mucedola) was provided as necessary and changed at least 2 times a week.
- Diet (e.g. ad libitum): ad libitum (laboratory rodent diet (4 RF 21,Mucedola S.r.l., Via G. Galilei, 4,
20019, SettimoMilanese (MI), Italy)
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 7 weeks
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C±2°C
- Humidity (%): 55%±15%
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12/12
IN-LIFE DATES: From: 01.03.2021 To: 14.04.2021
Administration / exposure
- Route of administration:
- oral: gavage
- Vehicle:
- corn oil
- Details on oral exposure:
- PREPARATION OF DOSING SOLUTIONS: The required amount of test item was suspended in the vehicle. The preparations were made daily at concentration of 25, 75 and 250 mg/mL, unless specified otherwise based on stability data stated in the validation study (section 3.4). Concentrations were calculated and expressed in terms of test item as supplied.
VEHICLE
- Amount of vehicle (if gavage): 4 ml/kg bw - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- Analysis was performed in a separate study in order to validate the analytical method and the preparation procedure and to verify the stability of the preparations (ERBC Study no. A4158). The analysis confirmed a 11 days stability at room temperature in the range from 25 to 250 mg/mL.
Samples of the preparations prepared during the current study (the first and the last week of treatment where possible) were analysed to check the homogeneity and concentration.
Chemical analysis was carried out by the Analytical Chemistry Department at ERBC. The validated software used for this activity was Analyst 1.6.2. Results of the analyseswere within the acceptability limits stated in ERBC SOPs for concentration of suspension (85-115%), - Duration of treatment / exposure:
- - Males: at least 28 days. The duration of treatment covered a minimum of 2 consecutive weeks prior to pairing and thereafter through the day before necropsy.
- Females: up to 60 days. The duration of treatment covered a minimum of 2 consecutive weeks prior to pairing and thereafter during pairing, post coitum and post partum periods until Day 13 post partum or the day before sacrifice. - Frequency of treatment:
- daily
Doses / concentrationsopen allclose all
- Dose / conc.:
- 0 mg/kg bw/day (actual dose received)
- Remarks:
- Test group 1 (control)
- Dose / conc.:
- 100 mg/kg bw/day (actual dose received)
- Remarks:
- Test group 2
- Dose / conc.:
- 300 mg/kg bw/day (actual dose received)
- Remarks:
- Test group 3
- Dose / conc.:
- 1 000 mg/kg bw/day (actual dose received)
- Remarks:
- Test group 4
- No. of animals per sex per dose:
- 10 animals per sex per dose
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- Dose levels have been selected in consultation with the Sponsor based on information from previous studies.
In a previous repeated-dose toxicity study (OECD 407) with a structural similar substance the NOAEL was considered to be 1000 mg/kg/day - Positive control:
- no
Examinations
- Observations and examinations performed and frequency:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Mortality: Throughout the study, all animals were checked early in each working day in the morning and in the afternoon. At weekends and Public Holidays a similar procedure was followed except that the final check was carried out at approximately mid-day.
- Clinical observations: Once before commencement of treatment and at least once daily during the study, each animal was observed and any clinical signs recorded. Observations were performed at the same time interval each day, the interval was selected taking into consideration the presence of post-dose reactions. All observations were recorded for individual animals.
DETAILED CLINICAL OBSERVATIONS: No
BODY WEIGHT: Yes
- Time schedule for examinations: once a week
- Males: weekly from allocation to termination.
- Females: at weekly interval from allocation, where possible, to positive identification of mating and on Days 0, 7, 14 and 20 post coitum.
- Dams were weighed on Days 1, 4, 7 and 13 post partum and just prior to necropsy.
FOOD CONSUMPTION: Yes
- Time schedule: weekly during the pre-mating period, starting Day 1 of dosing up to mating.
- Individual food consumption for the females were measured on Days 7, 14 and 20 post coitum starting from Day 0 post coitum and on Days 7 and 13 post partum starting from Day 1 post partum
WATER CONSUMPTION (if drinking water study): No
CLINICAL BIOCHEMISTRY: Yes
- Anaesthetic used for blood collection (for hormone determination) : Yes (Isoflurane)
- How many animals: all parental males and females/ Pups: from each litter (1 sample for males and 1 sample for females)
- Parameter checked: T4, TSH (serum) - Sacrifice and pathology:
- SACRIFICE
Parental animals that had completed the scheduled test period were killed by exsanguinations under isoflurane anaesthesia. One female of the low dose group was sacrificed for humane reasons, before the scheduled test period, under carbon dioxide asphyxiation.
The males were killed at completion of mating of all females (after 35 days of treatment). The females with live pups were killed on Day 14 post partum. The females with total litter loss were killed on the day of the occurrence of total litter loss or shortly after. One control female did not show evidence of copulation, however it gave birth. The females which did not give birth 25 days after positive identification of mating were killed shortly after.
GROSS NECROPSY
- Females: All females were also examined for the number of visible implantation sites (pregnant
animals). Uteri of females with no visible implantations were immersed in a 20% solution of ammonium sulphide to reveal evidence of implantation.
ORGAN WEIGHTS
- Parental animals: From all animals completing the scheduled test period, the following organs/tissues were dissected free of fat and weighed: Abnormalities, epididymides, liver, ovaries with oviducts, prostate gland (dorsolateral and ventral), seminal vesicles with coagulating glands, testes, thyroid gland, uterus – cervix, vagina.
- The ratios of organ weight to body weight was calculated for each animal.
TISSUE FIXATION
Samples from all the tissues/organs mentioned under “organ weights” were fixed and preserved in 10% neutral buffered formalin (except eyes, optic nerves, Harderian glands, ovaries, testes and epididymides which were fixed in modified Davidson’s fluid and preserved in 70% ethyl alcohol)
HISTOPATHOLOGY
- The following tissues were required for histopathological examination: Abnormalities, epididymides, liver, ovaries with oviducts, prostate gland (dorsolateral and ventral), seminal vesicles with coagulating glands, testes, thyroid gland, uterus – cervix, vagina.
- After dehydration and embedding in paraffin wax, sections of the tissues were cut at 5 micrometer thickness and stained with haematoxylin and eosin. In addition, the testes and epididymides were cut at 2-3 micrometer thickness and stained with Periodic Acid Schiff (PAS).
- The examination was restricted as described:
o Tissues mentioned in “organ weights” from all males and females in the control and high dose groups killed at term. The morphological evaluation of the seminiferous epithelium (staging of spermatogenic cycle) was also included.
o Tissues mentioned in “organ weights” from all animals killed or dying during the treatment period.
o All abnormalities in all groups.
- The examination of liver and thyroid was extended to animals of the low and mid-dose groups of both sexes based on treatment-related changes observed in the high dose group. - Statistics:
- Standard deviations were calculated as appropriate. For continuous variables, the significance of the differences amongst group means was assessed by Dunnett’s test or a modified t test, depending on the homogeneity of data. Statistical analysis of histopathological findings was carried out by means of the non-parametric Kolmogorov Smirnov test. The non-parametric Kruskal-Wallis analysis of variance (non-continuous variables) was used for the other parameters. Intergroup differences between the control and treated groups were assessed by the non-parametric version of theWilliams test. The criterion for statistical significance was p < 0.05. The mean values, standard deviations and statistical analysis were calculated from actual values in the computer without rounding off.
Results and discussion
Results of examinations
- Clinical signs:
- effects observed, non-treatment-related
- Description (incidence and severity):
- No signs were recorded in males throughout the study.
No treatment-related clinical signs were recorded in females before pairing. During gestation, signs of difficulty to delivery were recorded in one female at 100 mg/kg/day.
No treatment-related clinical signs were recorded during post partum phase. - Mortality:
- mortality observed, non-treatment-related
- Description (incidence):
- On gestation day 22, one female at 100 mg/kg/day was sacrificed following the occurrence of important clinical signs like prolapse of the uterus, pallor, piloerection and coldness to touch. The presence of these signs was related to difficulty in delivery.
One female at 300 mg/kg/day was found dead on gestation day 23. This female gave birth 8 pups on gestation day 22 (found dead) and additional 3 live pups were found in the cage on Day 23 post coitum. Both females had a prolonged time of gestation without completing the parturition and the impaired delivery was the reason of the humane sacrifice for the low dose female (100 mg/kg/day) and the occurrence of premature death for the mid-dose female (300 mg/kg/day). The gross and microscopic evaluation did not allow to establish the cause of impaired parturition. However, since these findings occurred in single animals, they could be considered unrelated to treatment. - Body weight and weight changes:
- no effects observed
- Food consumption and compound intake (if feeding study):
- no effects observed
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- not examined
- Haematological findings:
- not examined
- Clinical biochemistry findings:
- effects observed, treatment-related
- Description (incidence and severity):
- A dose-related increase in the TSH level was observed in parental males. The increase was 63% at 300 mg/kg/day and 113% at 1000 mg/kg/day, compared to controls. This finding could be related with the follicular cell hypertrophy of thyroid gland observed at histopathology and, due to the absence of T4 changes, could represent a subclinical hypothyroidism. Table included under "Any other information on results incl. tables".
- Urinalysis findings:
- not examined
- Behaviour (functional findings):
- not examined
- Immunological findings:
- not examined
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Description (incidence and severity):
- There were no treatment-related changes in terminal body weight when compared to controls.
Treatment-related increases in the liver and thyroid weights, absolute or relative to body weight, were observed at all dose levels with a dose relationship.
The statistically significant increase in absolute and relative thyroid weight for mid and high dose males (21% of mean relative weight, respectively) correlated microscopically with follicular cell hypertrophy of thyroid gland. In addition, statistically significant increase in absolute and relative mean liver weight for low, mid- and high dose males (15%, 20% and 30% of mean relative weight, respectively) and high dose females (23% of mean relative weight) was noted and for mid and high dose males and high dose females correlated microscopically with centrilobular hepatocellular hypertrophy.
Any organ weight changes other than those listed above were within the range of occasionally
observed and expected spontaneous changes in rats of the same age and considered unrelated to treatment.
Table included under "Any other information on results incl. tables". - Gross pathological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- There were no treatment-related macroscopic observations at the end of the treatment period. Any macroscopic observations had a comparable incidence in control and treated groups and/or are characteristically seen in untreated rats of the same age and were considered incidental and unrelated to treatment.
- Neuropathological findings:
- not examined
- Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Description (incidence and severity):
- Microscopic observations associated with the oral administration of the test item were present in the thyroid and liver.
- In the thyroid gland, treatment-related microscopic observations consisted in follicular cell hypertrophy which was characterised by the presence of increased size and height of follicular cells with diffuse distribution. Follicular lumen was decreased in size and filled with normal staining to pale colloid and the cytoplasm of the follicular cells was enriched with small clear vacuoles. The above mentioned finding was mild to marked in all males (10/10) and moderate in one female (1/10) of high dose group and of mild severity in males (3/10) of mid-dose group. In males at the mid and high dose, this finding correlated with
changes in thyroid related hormone levels measured in parental males (increased values of TSH) and with the increased absolute and relative thyroid mean weights. Follicular cell hypertrophy of the thyroid gland was considered treatment-related.
- In the liver, treatment-related microscopic observations consisted in centrilobular hepatocellular hypertrophy of
minimal to mild severity was present in all males (10/10) and most females (7/10) at the high
dose and of minimal severity in few males (2/10) of the mid-dose group. Hepatocyte hypertrophy
consisted of enlarged hepatocytes containing variable amounts of fine granular pale
eosinophilic (ground glass) cytoplasm.
- There were no microscopic observations in the
testes as per specific stage aware evaluation using PAS stain.
- Any microscopic observations other than those listed above had a comparable incidence in control and treated groups and/or are characteristically seen in untreated rats of the same age and were considered incidental and unrelated to treatment.
Effect levels
- Dose descriptor:
- NOAEL
- Effect level:
- 100 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
- Remarks on result:
- other: the relationship between thyroid hypertrophy and TSH higher level is not necessarily indicative of thyroid dysfunction since the thyroid hormone changes did not include low serum of T4
Target system / organ toxicity
- Critical effects observed:
- no
Any other information on results incl. tables
Table 1: Liver and thyorid weights
| Males | HCD | Females | HCD | |||||||
Dose level (mg/kg/day) | Control | 100 | 300 | 1000 |
| Control | 100 | 300 | 1000 |
| |
Liver | Abs. weight (g) | 12.282 | 14.248* | 14.946* | 16.092* | 14.52 | 12.234 | 12.608 | 13.924 | 15.227* | 11.29 |
Rel. weight (g) | 2.798 | 3.210* | 3.350* | 3.636* | 3.65 | 3.955 | 4.221 | 4.534 | 4.854* | 3.83 | |
Thyroid | Abs. weight (g) | 0.0230 | 0.0254 | 0.0284* | 0.0283* | 0.026 | 0.0221 | 0.0216 | 0.0238 | 0.0230 | 0.023 |
Rel. weight (g) | 0.0053 | 0.0057 | 0.0064 | 0.0064 | 0.006 | 0.0074 | 0.0072 | 0.0078 | 0.0074 | 0.008 | |
* = Statistical significant from control at p<0.05 |
Table 2: Thyroid hormone measurements in parental males and male and female pups
Parental Males (Group mean data) | |||||
| Group | ||||
Parameter/units | 1 | 2 | 3 | 4 | |
Thyroxine nmol/L | Mean SD N | 41.4 3.9 10 | 41.4 3.6 9 | 44.8 5.3 10 | 38.1 4.1 10 |
Thyroid Stimulating Hormone ng/mL | Mean SD N | 8.79 1.99 10 | 9.6 1 1.74 9 | 14.34+ 4.24 10 | 18.68+ 9.13 10 |
Controls from group(s): 1 Subgroup(s): 1 * = mean value of group is significantly different from control at p < 0.05 + = mean value of group is significantly different from control at p < 0.01 | |||||
Pups Day14 Post Partum_Males (Group mean data) | |||||
| Group | ||||
Parameter/units | 1 | 2 | 3 | 4 | |
Thyroxine nmol/L | Mean SD N | 54.9 10.1 7 | 52.3 6.5 7 | 52.5 6.9 8 | 53.3 2.8 9 |
Thyroid Stimulating Hormone ng/mL | Mean SD N | 3.80 0.65 7 | 3.60 0.68 7 | 3.83 0.72 8 | 3.67 0.85 9 |
Controls from group(s): 1 Subgroup(s): 1 * = mean value of group is significantly different from control at p < 0.05 + = mean value of group is significantly different from control at p < 0.01 | |||||
Pups Day14 Post Partum_Females (Group mean data) | |||||
| Group | ||||
Parameter/units | 1 | 2 | 3 | 4 | |
Thyroxine nmol/L | Mean SD N | 56.0 7.6 7 | 55.3 10.5 7 | 55.6 8.9 8 | 54.8 8.0 9 |
Thyroid Stimulating Hormone ng/mL | Mean SD N | 3.54 0.53 7 | 3.71 0.53 7 | 4.33 0.89 8 | 3.67 0.76 9 |
Controls from group(s): 1 Subgroup(s): 1 * = mean value of group is significantly different from control at p < 0.05 + = mean value of group is significantly different from control at p < 0.01 |
Applicant's summary and conclusion
- Conclusions:
- In conclusion after repeated dose, treatment-related effects were seen in parental animals at 300 and 1000 mg/kg/day characterized by morphological changes in thyroid and liver tissues (hypertrophy) as well as by TSH alteration in males (increase). Nevertheless, the relationship between thyroid hypertrophy and TSH higher level is not necessarily indicative of thyroid dysfunction since the thyroid hormone changes did not include low serum of T4. No adverse effects were observed regarding developmental and reproductive parameters.
Based on these results, the NOAEL (No Observed Adverse Effect Level) can be conservatively
set at 100mg/kg/day for general repeated dose toxicity. However, no clear indications of
adverse effects were seen at the higher dose levels. - Executive summary:
For more details please refere to sextion 7.8.1
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.