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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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

Toxicological information

Developmental toxicity / teratogenicity

Currently viewing:

Administrative data

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From July 04, 1990 to September 17, 1990
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
test procedure in accordance with national standard methods

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1990
Report date:
1990

Materials and methods

Principles of method if other than guideline:
Time pregnant CD (Sprague-Dawley) rats, 25 sperm-positive females per group, were exposed to the test substance by gavage once daily on Gestation Day (GD) 6 to 15 at 0, 500, 1000 or 2000 mg/kg bw/day in corn oil. Clinical observations were recorded, as well as maternal bodyweight and food consumption. At sacrifice on GD 20, the dams were evaluated for bodyweight, liver and gravid uterine weight. Ovarian corpora lutea were counted and the status of uterine implantation sites (i.e. resorptions, dead fetuses, live fetuses) was recorded. All fetuses were dissected from the uterus, counted, weighed, sexed and examined for external abnormalities. Approximately one half of the live fetuses in each litter were examined for visceral malformations and variations. The heads were fixed and examined for soft tissue craniofacial malformations and variations. Intact fetuses were examined for skeletal malformations and variations.
GLP compliance:
yes
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
2,2'-[[5-acetamido-4-[(2-bromo-4,6-dinitrophenyl)azo]-2-methoxyphenyl]imino]diethyl diacetate
EC Number:
222-813-1
EC Name:
2,2'-[[5-acetamido-4-[(2-bromo-4,6-dinitrophenyl)azo]-2-methoxyphenyl]imino]diethyl diacetate
Cas Number:
3618-72-2
Molecular formula:
C23H25BrN6O10
IUPAC Name:
2-{[2-(acetyloxy)ethyl]({4-[ (E)-2-(2-bromo-4,6-dinitrophenyl)diazen-1-yl]-5acetamido-2-methoxyphenyl})amino}ethyl acetate
Test material form:
solid
Remarks:
Press cake
Details on test material:
Storage: room temperature
Different stock solutions in acetone and acetonitrile were prepared and stored in the refrigerator. Subsequent dilutions were used in the preparation of quality control fortification samples.

Test animals

Species:
rat
Strain:
Sprague-Dawley
Remarks:
CD
Details on test animals or test system and environmental conditions:
From: Charles River Laboraties
Number of females: 100 sperm-positive female rats (4 groups of 25 animals)
Age: ca. 10 weeks
Weight: 223 - 268g
Food: no. 5002 Purina Certified Rodent Chow and Water: deionized/filtered tap water ad libitum
Temperature: ca. 23°C
Humidity: 40 - 70%
Light: 12:12h light:dark cycle

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
Pregnant females were exposed to the test substance, by gavage once daily from gestational days 6 through 15 at doses of 0, 500, 1000 or 2000 mg/kg bw/day in corn oil (based on a range-finding study). The dosing volume was 10 mL/kg bw and was adjusted based on each animal's most recent body weight.
Analytical verification of doses or concentrations:
yes
Remarks:
HPLC (Waters 840 HPLC and data system)
Details on analytical verification of doses or concentrations:
Prior to formulation of study dosing suspensions, aliquots of formulations of test substance in vehicle, bracketing the range of concentrations to be used in this used were assayed for homogeneity and stability. Suspension concentrations were also verified for all dose levels. Dosing formulations were homogeneous and stable for at least 28 days and the concentrations were within 90-110% of the nominal concentrations. The content of test substance in the vehicle was below the detection limit.
Triplicate 1.0 mL samples of the vehicle solution (corn oil) and of the dosing formulations, 50.0, 100.0 and 200.0 mg/mL were analysed.
Details on mating procedure:
For breeding, individual females were placed in the home cage of singly-housed males. On the following morning and each morning thereafter, the females were examinated for the presence of vaginal sperm (considered as GD 0). Sperm-positive females were individually housed until scheduled sacrifice on GD 20.
Duration of treatment / exposure:
From GD 6 to 15
Frequency of treatment:
Once daily
Duration of test:
Until GD 20
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day (actual dose received)
Dose / conc.:
500 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
Dose / conc.:
2 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
25
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale:
The doses were 0, 500, 1000 and 2000 mg/kg/day based on a rangefinding study in pregnant rats. The highest dose level was chosen to induce overt maternal toxicity, but not to cause a weight loss greater than 20% when compared to concurrent controls, nor to cause greater than 10% maternal mortality. The low dose was selected to be a maternal/developmental No Observable Adverse Effect Level (NOAEL). The mid dose was one-half the high dose and twice the low dose.

Examinations

Maternal examinations:
- Clinical observations: daily and twice daily during the dosing period (1-2h after dosing period)
- Maternal body weight: on gestational days 0, 6, 9, 12, 15, 18 and 20
- Food consumption: gestational days 0-6, 6-9, 9-12, 12-15, 15-18 and 18-20
Ovaries and uterine content:
- At scheduled sacrifice on gestational day 20, the dams were evaluated for body weight, liver and gravid uterine weight
- Ovarian corpora lutea were counted and the status of uterine implantation sites (i.e., resorptions, dead fetuses, live fetuses, pre and post-implantations) was recorded.
Fetal examinations:
- Fetuses were dissected from the uterus, counted, weighted, sexed and examined for external abnormalities as well as for visceral malformations and variations.
- Sections of the heads were examined for soft tissue craniofacial malformations and variations.
- All fetuses were eviscerated, fixed in alcohol, and stained with alizarin red S/alcian blue.
- Intact fetuses were examined for skeletal malformations and variations.
Statistics:
Appropriate General Linear Models procedures were used for the analysis of variances ANOVA + Bartlett's test for homogeneity of variance
Dunnett's Multiple Comparison Test
One-tailed or two-tailed tests were used depending on the endpoints
Chi-Square Test for Independance and Test for linear Trend on proportions
Fisher's Exact Probability Test
Indices:
Gestational parameters (ovarian corpora lutea, uterine implantation sites, resorptions, dead fetuses, live fetuses, pre and post-implantation losses)

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
- Green, dark and/or green feces at 500 mg/kg bw/day and above
- Piloerection at all concentrations (GD 12-20)
- Vaginal bleeding at 2000 mg/kg bw/day in one dam on GD 15
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Body weight loss in 5, 2 and 2 dams in the groups 500, 1000 and 2000 mg/kg bw/day (GD 7 - 10)
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
no effects observed
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined

Maternal developmental toxicity

Number of abortions:
no effects observed
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
effects observed, non-treatment-related
Description (incidence and severity):
Two litters at 500 mg/kg bw/day were fully resorbed; all remaining pregnant animals had one or more live fetuses on GD 20
Early or late resorptions:
effects observed, non-treatment-related
Dead fetuses:
no effects observed
Description (incidence and severity):
The numbers of litters evaluated were 11-14 per groups
Changes in pregnancy duration:
not examined
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed
Changes in number of pregnant:
no effects observed
Other effects:
effects observed, non-treatment-related
Description (incidence and severity):
Pregnancy rate was unexpectedly low (44-56%) and approximately equivalent across all groups

Effect levels (maternal animals)

Key result
Dose descriptor:
NOAEL
Effect level:
> 2 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Remarks on result:
not determinable due to absence of adverse toxic effects

Maternal abnormalities

Key result
Abnormalities:
no effects observed

Results (fetuses)

Fetal body weight changes:
no effects observed
Description (incidence and severity):
Just slightly reduced at 2000 mg/kg bw/day with no obvious dose-related trends
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): 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
Changes in postnatal survival:
not examined
External malformations:
no effects observed
Description (incidence and severity):
One fetus in each treatment group exhibited external malformations
Skeletal malformations:
effects observed, non-treatment-related
Description (incidence and severity):
Some malformations were observed in 0-3 fetuses in each group with no treatment-related patterns of incidence or severity.
Visceral malformations:
no effects observed
Details on embryotoxic / teratogenic effects:
No effects observed

Effect levels (fetuses)

Key result
Dose descriptor:
NOAEL
Effect level:
> 2 000 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
Treatment related:
no

Applicant's summary and conclusion

Conclusions:
Under the study conditions, the maternal and developmental NOAELs for the test substance administered by gavage from GD 6 to 15 were determined to be >2000 mg/kg bw/day (i.e. 1992 mg a.i./kg bw/day).
Executive summary:

A study was conducted to determine the developmental toxicity of the test substance (99.61% purity). in rat. Female Sprague Dawley rats (25/dose) were administered by gavage the following concentrations (dissolved corn oil, at a volume of 10 mL/kg bw): 0, 500, 1000 and 2000 mg/kg bw/day from gestational day (GD) 6 to 15. The homogeneity, stability and concentrations of dosing suspensions were analysed and were within acceptable values. Clinical signs were recorded daily and twice daily during the dosing period. Maternal body weight were measured on GD 0, 6, 9, 12, 15, 18 and 20 and food consumption on GD 0-6, 6-9, 9-12, 12-15, 15-18 and 18-20. At scheduled sacrifice on GD 20, the dams were evaluated for body weight, liver and gravid uterine weight. Ovarian corpora lutea were counted and the status of uterine implantation sites (i.e., resorptions, dead fetuses, live fetuses, pre and post-implantations) was recorded. Fetuses were dissected from the uterus, counted, weighted, sexed and examined for external abnormalities as well as for visceral malformations and variations. Sections of the heads were examined for soft tissue craniofacial malformations and variations. All fetuses were eviscerated, fixed in alcohol, and stained with alizarin red S/alcian blue. Intact fetuses were examined for skeletal malformations and variations. Except for green, dark and/or green feces at 500 mg/kg bw/day and above, piloerection at all concentrations (GD 12-20) and vaginal bleeding at 2000 mg/kg bw/day in one dam on GD 15, no other effects were observed. Gestational parameters (e.g. ovarian corpora lutea, uterine implantation sites, resorptions, dead fetuses, live fetuses, pre and post-implantation losses) were not modified by the treatment. Two litters at 500 mg/kg bw/day were fully resorbed; all remaining pregnant animals had one or more live fetuses on GD 20. Fetal body weight was slightly reduced at 2000 mg/kg bw/day with no obvious dose-related trends. Malformations (external and skeletal) were recorded but with no treatment-related patterns of incidence or severity. Under the study conditions, the maternal and developmental NOAELs were determined to be >2000 mg/kg bw/day (i.e. 1992 mg a.i./kg bw/day) (Rochelle, 1990).