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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 226-107-4 | CAS number: 5280-80-8
- 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
Developmental toxicity / teratogenicity
Administrative data
- Endpoint:
- developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 25. Nov 2021 - 31. Mar 2022
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Cross-reference
- Reason / purpose for cross-reference:
- reference to same study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 022
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- other: OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
- Version / remarks:
- 29 July 2016
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
Test material
- Reference substance name:
- 3,3'-[(2-chloro-5-methyl-p-phenylene)bis[imino(1-acetyl-2-oxoethylene)azo]]bis[4-chloro-N-(3-chloro-o-tolyl)benzamide]
- EC Number:
- 226-970-7
- EC Name:
- 3,3'-[(2-chloro-5-methyl-p-phenylene)bis[imino(1-acetyl-2-oxoethylene)azo]]bis[4-chloro-N-(3-chloro-o-tolyl)benzamide]
- Cas Number:
- 5580-57-4
- Molecular formula:
- C43H35Cl5N8O6
- IUPAC Name:
- 3,3'-{(2-chloro-5-methyl-1,4-phenylene)bis[imino(1,3-dioxobutane-2,1-diyl)diazene-2,1-diyl]}bis[4-chloro-N-(3-chloro-2-methylphenyl)benzamide]
- Test material form:
- solid: nanoform
- Details on test material:
- Physical state/ appearance: yellow powder
Constituent 1
- Specific details on test material used for the study:
- Batch no.: 0004460012
Purity: 99.3 wt%
Expiry/Retest date: 18 Jan 2031
Appearance: Yellow powder
Storage conditions: Room temperature
The determination of the identity, strength, purity, composition and stability of the test item was the responsibility of the Sponsor. The Sponsor declared that the characterisation of the test item was carried out according to a quality system.
Test animals
- Species:
- rat
- Strain:
- Wistar
- Remarks:
- Wistar Hannover
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River Germany, D-97633 Sulzfeld
- Females nulliparous and non-pregnant: yes
- Age at study initiation: (P) 8-10 weeks
- Weight at study initiation: (P) Males: 208-227 g; Females: 198 -217 g
- Housing: 5 of one sex to a cage until pairing; 1:1 (m/f) during pairing; after pairing: females individually, males as before mating
- Diet: ad libitum, laboratory rodent diet (4 RF 21,Mucedola S.r.l., Via G. Galilei, 4, 20019 SettimoMilanese (MI), Italy)
- Water: ad libitum
- Acclimation period: 2 weeks
ENVIRONMENTAL CONDITIONS
- Temperature: 22°C ± 2°C
- Humidity: 55% ± 15%
- Air changes (per hr): 15 to 20
- Photoperiod (hrs dark / hrs light): 12/12
IN-LIFE DATES: From: 25 Nov 2021 To: Feb 2022 (females), Jan 2022 (males)
Administration / exposure
- Route of administration:
- oral: gavage
- Vehicle:
- CMC (carboxymethyl cellulose)
- Remarks:
- 0.5 %
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS:
The required amount of test item was suspended in the vehicle. The preparations were made weekly and every three days at concentration of 10, 30 and 100 mg/mL. Concentrations were calculated and expressed in terms of test item as supplied.
VEHICLE
- Amount of vehicle (if gavage): 10 mL/kg bw - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- The analytical method was validated in the range from 10 to 100 mg/mL. Linearity, accuracy and precision were within the limits stated in the validation protocol (r > 0.99; accuracy 85-115%; precision CV < 10%).
A 28 hour stability at room temperature and an 8 day stability at 2-8°C were verified in the range from 10 to 100 mg/mL.
The proposed preparation procedure for the test item was checked in the range from 10 to 100 mg/mL by chemical analysis (concentration and homogeneity) to confirmthat the method was suitable. Final results for all levels were within the acceptability limits stated in internal SOPs for concentration (85-115%) and homogeneity (CV < 10%).
Samples of the formulations prepared on the first and the last week were analysed to check the homogeneity and concentration. Results of the analyses were within the acceptability limits stated in internal SOPs for suspensions (85-115% for concentration and CV < 10% for homogeneity).
Chemical analysis was carried out by the Analytical Chemistry Department at ERBC. - Details on mating procedure:
- - M/F ratio per cage: 1:1
- Length of cohabitation: 14 d
- Proof of pregnancy: spermidentification, vaginal plug in situ or copulation plugs found in the cage tray
- After successful mating each pregnant female was caged: individually - Duration of treatment / exposure:
- Males: 34/35 d
Females: 66 d - Frequency of treatment:
- once daily
Doses / concentrationsopen allclose all
- Dose / conc.:
- 0 mg/kg bw/day (actual dose received)
- Remarks:
- Vehicle control
- Dose / conc.:
- 100 mg/kg bw/day (actual dose received)
- Remarks:
- Low dose group
- Dose / conc.:
- 300 mg/kg bw/day (actual dose received)
- Remarks:
- Mid dose group
- Dose / conc.:
- 1 000 mg/kg bw/day (actual dose received)
- Remarks:
- High dose group
- No. of animals per sex per dose:
- 10
- Control animals:
- yes, concurrent vehicle
Examinations
- Maternal examinations:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily (mortality), at least once daily (clinical signs)
- Cage side observations: mortality, clinical signs
BODY WEIGHT: Yes
- Time schedule for examinations: weekly before mating, on days 0, 7, 14 and 20 p.c.; on days 1, 4, 7 and 13 p.p. and prior necropsy
FOOD CONSUMPTION AND COMPOUND INTAKE:
The weight of food consumed by each cage and females was recorded weekly during the pre-mating period starting from the beginning of treatment up to mating. Individual food consumption for mated 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.
PARTUITION CHECK:
- three times a day during the working day and twice daily during the weekends and Public Holidays
- from Day 20 to Day 25 post coitum
OESTROUS CYCLE:
The assessment of oestrous cycles, by vaginal smears, was performed once daily in the morning in stock females and in allocated females for 2 weeks before the start of dosing.
IMMUNOANALYSIS:
- Thyroid hormone determination (T4 and TSH)
POSTMORTEM EXAMINATIONS:
- The clinical history of the males and females of the parental generation was studied and a detailed post mortem examination was conducted (including examination of the external surface and orifices). Changes were noted, the requisite organs weighed and the required tissue samples preserved in fixative and processed for histopathological examination.
- All females were also examined for the number of visible implantation sites (pregnant animals). Uteri from females with no visible implantations, and uterus from one female with implantations only on the right horn, were immersed in a 20% solution of ammonium sulphide to reveal evidence of implantation.
- The tissues indicated in Table 1 were prepared for microscopic examination and weighed, respectively. - Ovaries and uterine content:
- The ovaries and uterine content was examined after termination: No
- Blood sampling:
- As part of the necropsy procedure, blood samples (approximately 0.8 mL) were withdrawn under isoflurane anaesthesia from the abdominal vena cava.
- Fetal examinations:
- STANDARTISATION OF LITTER:
- on Day 4 post partum,
- litter size was adjusted by eliminating extra pups by random selection to yield, as nearly as possible, four pups per sex per litter
- Partial adjustment (for example, 5 males and 3 females) was acceptable
- no pups were eliminated when litter size dropped below the culling target (8 pups/litter).
PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
- number and sex of pups
- stillbirths, live births
- weight gain (weighing on day 1, 4, 7, 13)
- postnatal mortality
- presence of gross anomalies
- anogenital distance (AGD)
- pup weight on the day of AGD
- presence of nipples/areolae in male pups (Day 13/14)
GROSS EXAMINATION OF DEAD PUPS
BLOOD COLLECTION:
- On Days 4 and 14 post partum, as part of the necropsy procedure, blood samples of approximately 0.5 mL (per sex) were taken from each litter (1 sample for males and 1 sample for females, when possible). Blood samples were withdrawn under light ether anaesthesia from the heart (intracardiac puncture).
In order to obtain the required volume, blood samples were pooled from two or three pups on Day 4 post partum.
IMMUNOANALYSIS:
- Thyroid hormone determination (T4 and TSH) - 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.
- Indices:
- Gestation length was calculated as the time between the day of successful mating (Day 0 post coitum) and the day when the parturition is defined complete (Day 0 post partum).
Males:
Copulatory Index (%) = no. of males with confirmed mating/no. of males cohabitated ×100
Fertility Index (%) = no. of males which induced pregnancy/no. of males cohabitated ×100
Females:
Copulatory Index (%) = no. of females with confirmed mating/no. of females cohabitated ×100
Fertility Index (%) = no. of pregnant females/no. of females cohabitated ×100
Pup loss at Day 0 post partum was calculated as a percentage from the formula:
(Total litter size−Live litter size)/Total litter size ×100
Pre-natal loss was calculated as a percentage from the formula:
(no. of visible implantations−Live litter size at birth)/no. of visible implantations ×100
Post-natal loss at Day 4 post partum (before culling) was calculated as a percentage from
the formula:
(Live litter size at birth−live litter size at Day 4 (before culling))/Live litter size at birth ×100
Post-natal loss at Day 13 post partum (after culling) was calculated as a percentage from
the formula:
(Live litter size onDay 4 (after culling)−Live litter size onDay 13)/Live litter size onDay 4 (after culling) ×100
Males and females:
Pre coital Interval = The number of nights paired prior to the detection of mating - Historical control data:
- T4 and TSH historical data for Wistar Hannover rat in serum were available
Results and discussion
Results: maternal animals
General toxicity (maternal animals)
- Clinical signs:
- no effects observed
- Mortality:
- no mortality observed
- Body weight and weight changes:
- effects observed, non-treatment-related
- Description (incidence and severity):
- No differences in body weight were recorded between treated and control groups throughout the entire study.
A degree of variability in body weight gain was observed in both males and females over the course of the study and among all groups, but no relationship with the test item treatment was observed. Notably, there was a -58% decrease in body weight gain in high dose females at Day 7 post partum, compared to the control, but without statistical significance.
Being isolated event and without a dose-related trend, this case was not considered to be treatment related. - 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:
- not examined
- Endocrine findings:
- no effects observed
- Urinalysis findings:
- not examined
- Behaviour (functional findings):
- not examined
- Immunological findings:
- not examined
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Gross pathological findings:
- no effects observed
- Neuropathological findings:
- not examined
- Histopathological findings: non-neoplastic:
- no effects observed
- Histopathological findings: neoplastic:
- no effects observed
Maternal developmental toxicity
- Number of abortions:
- no effects observed
- Pre- and post-implantation loss:
- not examined
- Total litter losses by resorption:
- not examined
- Early or late resorptions:
- not examined
- Dead fetuses:
- effects observed, non-treatment-related
- Changes in pregnancy duration:
- no effects observed
- Changes in number of pregnant:
- no effects observed
Effect levels (maternal animals)
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- >= 1 000 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Remarks on result:
- not determinable due to absence of adverse toxic effects
Results (fetuses)
- Fetal body weight changes:
- no effects observed
- Reduction in number of live offspring:
- no effects observed
- Changes in sex ratio:
- no effects observed
- Changes in litter size and weights:
- no effects observed
- Anogenital distance of all rodent fetuses:
- no effects observed
- Changes in postnatal survival:
- no effects observed
- External malformations:
- no effects observed
- Skeletal malformations:
- not examined
- Visceral malformations:
- not examined
- Other effects:
- effects observed, non-treatment-related
- Description (incidence and severity):
- A statistical significance decrease in mean thyroid weight was found in male pups of mid-dose group respect to than from control dams. This decrease (-21%) was slight and not dose related, therefore considered incidental. No other statistically significant changes were noted.
Effect levels (fetuses)
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- >= 1 000 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Remarks on result:
- not determinable due to absence of adverse toxic effects
Fetal abnormalities
- Key result
- Abnormalities:
- no effects observed
Overall developmental toxicity
- Key result
- Developmental effects observed:
- no
Any other information on results incl. tables
Table 2: Reproduction data
Groups | 1 | 2 | 3 | 4 |
Paired No. | 10 | 10 | 10 | 10 |
Positive identification of mating No. | 9 | 10 | 10 | 10 |
Not pregnant | 3 | 0 | 1 | 1 |
Conceiving number | 7 | 10 | 9 | 9 |
Total resorptions | 0 | 0 | 1 | 0 |
Total litter loss | 0 | 0 | 0 | 0 |
Positive identification of mating 1 - 5 days | 9 | 9 | 8 | 7 |
Positive identification of mating 6 - 14 days | 0 | 1 | 2 | 3 |
No. of females with live pups on Day 14 post partum | 7 | 10 | 8 | 9 |
Table 3: Reproduction data and litter data - group means
Summary of Reproduction Data | ||||
| Dosage levels mg/kg/day | |||
Observations | 0 | 100 | 300 | 1000 |
No. Dams Inseminated | 9 | 10 | 10 | 10 |
Oestrus cycle (mean number) | 4 | 3 | 4 | 4 |
No. Dams Pregnant | 7 | 10 | 9 | 9 |
Percent Dams Pregnant | 70.0 | 100.0 | 90.0 | 90.0 |
Copulatory interval (1-5 days) | 9 | 9 | 8 | 7 |
Copulatory interval (6-14 days) | 0 | 1 | 2 | 3 |
No. Dams Died During Study | 0 | 0 | 0 | 0 |
No. Dams with Resorptions only | 0 | 0 | 1 | 0 |
No. of Litters | 7 | 10 | 8 | 9 |
Total No. Implantation | 83 | 101 | 88 | 104 |
Mean No. Implants/Pregnancy | 11.9 | 10.1 | 9.8 | 11.6 |
Pre natal Loss (%) | 4.3 | 10.0 | 5.6 | 9.5 |
Post natal Loss (%) | 0.0 | 1.3 | 0.0 | 0.0 |
|
|
|
|
|
Summary of Litter data | ||||
| Dosage levels mg/kg/day | |||
| 0 | 100 | 300 | 1000 |
Pairs started (no.) | 10 | 10 | 10 | 10 |
Dams with live pup at birth (no.) | 7 | 10 | 8 | 9 |
Dams with live pups at Day 4 pp (no.) | 7 | 10 | 8 | 9 |
Live litter size at birth (mean) | 11.3 | 9.00 | 10.1 | 10.7 |
Live litter size at Day 4 (mean) | 11.3 | 9.00 | 9.6 | 10.7 |
Sex ratio (%) at birth (mean) | 47.9 | 54.5 | 52.4 | 48.9 |
Sex ratio (%) at Day 4 (mean) | 47.9 | 54.5 | 52.8 | 48.9 |
Litter weight at Day 1 (mean) | 78.7 | 64.3 | 69.3 | 73.8 |
Litter weight at Day 4 (mean) | 124.1 | 100.9 | 110.3 | 113.6 |
Pup weight at Day 1 (mean) | 7.0 | 7.2 | 7.4 | 7.0 |
Pup weight at Day 4 (mean)
| 11.1 | 11.4 | 11.9 | 10.9 |
Pup weight at the Day of AGD (Day 1pp) |
|
|
|
|
Mean males | 7.30 | 7.30 | 7.30 | 7.30 |
Mean females | 6.81 | 6.81 | 6.81 | 6.81 |
Pup AGD normalized (Day 1pp) |
|
|
|
|
Mean males | 2.23 | 2.23 | 2.23 | 2.23 |
Mean females | 1.23 | 1.23 | 1.23 | 1.23 |
Male pup nipple present at Day 13 (no.) | 0 | 0 | 0 | 0 |
Pup weight at Day 13 (mean) | 31.8 | 32.6 | 33.5 | 31.2 |
Pups with major abnormalities at necopsy |
|
|
|
|
Dams with 0 | 7 | 10 | 8 | 9 |
Dams with 1 | 0 | 0 | 0 | 0 |
Dams with > 2 | 0 | 0 | 0 | 0 |
Pre-natal loss (implantations minus live births)(no.) |
|
|
|
|
Females with 0 | 4 | 6 | 5 | 4 |
Females with 1 | 2 | 1 | 2 | 3 |
Females with 2 | 1 | 1 | 1 | 1 |
Females with > 3 | 0 | 2 | 1 | 1 |
Post-natal (live births minus live at Day 1) (no.) |
|
|
|
|
Females with 0 | 7 | 10 | 6 | 9 |
Females with 1 | 0 | 0 | 1 | 0 |
Females with 2 | 0 | 0 | 1 | 0 |
Females with > 3 | 0 | 0 | 0 | 0 |
Post-natal (live births at Day 4 after culling minus live Day 14) (no.) |
|
|
|
|
Females with 0 | 7 | 9 | 8 | 9 |
Females with 1 | 0 | 1 | 0 | 0 |
Females with 2 | 0 | 0 | 0 | 0 |
Females with > 3 | 0 | 0 | 0 | 0 |
Applicant's summary and conclusion
- Conclusions:
- The NOAEL for general toxicity, reproductive and developmental toxicity was considered to be 1000 mg/kg/day, for male and female parental animals and pups.
- Executive summary:
This screening study for reproductive and developmental toxicity was conducted according to OECD Guideline 421 and GLP. The effects of the test item on male and female reproductive performance, such as gonadal function, mating behaviour, conception, development of conceptuses, parturition and lactation of the offspring were investigated, upon daily administration of the test item by oral gavage toWistar Hannover rats.
Males were treated 14 days before pairing, trough the pairing period and up to one day before sacrifice for a total of 34/35 days. Femaleswere treated 14 days before pairing, trough pairing and gestation periods, and during post partum phase until Day 13 or the day before sacrifice (for a maximum of 66 days of treatment).
The doses used were: 0, 100, 300 and 1000 mg/kg/day and were administered at a dose volume of 10 mL/kg body weight. The control group received the vehicle alone (0.5% carboxymethyl cellulose).The following investigations were performed on parental animals: mortality, clinical signs, body weight, body weight gain, food consumption, oestrous cycle, mating performance, litter data, sex ratios, thyroid hormones determination (parental males), macroscopic observations and organ weights. Histopathological evaluation was performed on all males and females from control and high dose groups and on the abnormalities detected during post mortem examination. The identification of the stages of the spermatogenic cycle was also performed from all males in the control and high dose groups. Clinical signs, measurement of anogenital distance, post mortem external and/or internal examination were recorded for pups. Thyroid hormone levels were also determined in 1 pup/sex/group randomly selected at Day 14 post partum.
During the study, no mortality or other adverse effects concerning systemic toxicity, reproductive performance or development could be observed.
On the basis of the results obtained, no changes that could be clearly attributable to the treatment with the test item were observed at any dose level investigated. Therefore, the NOAEL (No Observed Adverse Effect Level) for general toxicity, reproductive and developmental toxicity was considered to be 1000 mg/kg/day, for male and female parental animals and pups.
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.