Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

A developmental/teratogenicity study without adverse effects is available for a structural analogue, and is offered in section 7.8.2. Hence, a reproductive/developmental screening study has not to be performed according to Column 2 of REACH Annex VIII. Furthermore, no effects were seen on reproductive organs in the repeat dose studies in structural analogues, and the category of substance (reactive dyes) is not known for reproductive toxicity effects. On the basis of animal welfare it is proposed that the developmental/teratogenicity study in conjunction with the lack of effects noted in the other toxicity studies is suitable to address this endpoint.


Short description of key information:
The absence of adverse effects on gonads in the repeat dose study together with the absence of adverse effects in a pre-natal development toxicity study and the overall absence of reproductive toxicity effects of the category of substance (reactive dyes) suggests that the test substance does not have reproductive toxicity potential.

Effects on developmental toxicity

Description of key information
No embryotoxic or teratogenic effects were observed with the structural analogue in the pre-natal development toxicity study.
Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
14. Sep to 25 Nov 1993
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
GLP compliance:
yes
Limit test:
yes
Species:
rat
Strain:
Wistar
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Hoechst AG
- Age at study initiation: 65 to 75 days
- Weight at study initiation: 193.1 +/- 14.2
- Fasting period before study: NA
- Housing: single
- Diet: Altromin 1310 ad libitum
- Water: tap water ad libitum
- Acclimation period: 7 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 22°C
- Humidity (%): 48 to 61%
- Air changes (per hr): 16 to 20
- Photoperiod (hrs dark / hrs light): 12/12


IN-LIFE DATES: From: 14. Sep To: 25. Nov 1993
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:

- Frequency of preparation: daily
- Administration: within 3 hours after preparation


VEHICLE
- Concentration in vehicle: 200 mg/kg nominal
- Amount of vehicle (if gavage): 5 ml/kg
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
-
Details on mating procedure:
- Impregnation procedure: cohoused
- M/F ratio per cage: 1:1
- Length of cohabitation: over night (3:30 pm to 7:30 am next day)
- Verification of same strain and source of both sexes: yes - own breeding facility
- Proof of pregnancy: sperm in vaginal smear, referred to as day 1 of pregnancy
Duration of treatment / exposure:
7. - 16. day of pregnancy
Frequency of treatment:
daily
Duration of test:
cesarean section on Day 21 of pregnancy
Remarks:
Doses / Concentrations:
1000 mg/kg bw
Basis:
nominal in water
No. of animals per sex per dose:
20 to 24 mated females
Control animals:
yes
Details on study design:
- Dose selection rationale: test item was tolerated in the acute and subacute studies without adverse effects
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE : Yes
- Time schedule for examinations: weekly
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/100 g body weight/day: Yes


POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 21
- Organs examined: all organs examined macroscopically
uterus - live and dead fetuses, resorption sites, placentas
ovaries - corpora lutea


OTHER:
- diameter of conceptuses undergoing resorption
- placenta weights
- presence of iron in uterus walls with ammonium sulphide to detect invisible implantation sites
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No
- Placenta weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
Fetal examinations:
- External examinations: Yes: all per litter
- Body weight: Yes: all per litter
- Crown-rump length: Yes: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: No data
Statistics:
comparison to actual control group and historical controls
MANOVA: body weight development, fetal weight, placental weight
PURI&SEN rank order test: food intake
Mantel-Haenszel's chi-squared test: live fetuses, intrauterine fetal death, number of implants, number of corpora lutea
multivariate analysis of variance: litter means of fetal weights, crown-rump length, placental weights
Fisher test: autopsy findings, body cross-sections, skelettal examination

Dams which had no live fetuses were excluded from the calculation of mean values and statistical evaluation
Indices:
No data
Historical control data:
Yes
Details on maternal toxic effects:
Maternal toxic effects:no effects

Details on maternal toxic effects:
black discoloration of feces and blue discoloration of urine due to excretion of dye
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: other:
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
only minor anomalies or variation within the historical range of spontaneous findings were observed
Abnormalities:
not specified
Developmental effects observed:
not specified
Conclusions:
No adverse effects were observed after daily administration of 1000 mg/kg/day Reactive Black 5 in dams or their fetuses.
Maternal NOAEL: 1000 mg/kg/day
Fetal NOEL: 1000 mg/kg/day
Executive summary:

In this limit test, Reactive Black 5, dissolved in distilled water, was administered orally by stomach tube in a single daily dose of 1000 mg/kg body weight to a group of 20 pregnant female Wistar rats from the 7th - 16th day of pregnancy. A simultaneous control group of the same size received the vehicle without test compound. On the 21st day of pregnancy, the dams were killed and delivered by caesarean section. The foetuses delivered by caesarean section were then examined morphologically for developmental disorders.

The studie showed that the repeated oral administration of Reactive Black 5, at a dose of 1000 mg/kg body weight in the sensitive phase of organogenesis for the conceptuses, did not lead to any impairment of the general physical condition of the dams or impaired intrauterine development of conceptuses.

The morphological examination of the foetuses with regard to stage of development, outwardly detectable anomalies as well as anomalies of the internal organs and the skeleton showed no indication of an embryotoxic or teratogenic effect of the compound. The findings observed are to be regarded as spontaneous in origin.

On the basis of the results of this limit test, the “no observed adverse effect level” for Reactive Black 5 in rats following oral administration lies at 1000 mg/kg body weight with regard to maternal and embryofoetal toxicity and teratogenicity.

No teratogenic effect was observed.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Species:
rat
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

In a limit test, the test item, Reactive Black 5, was administered orally on Days 7 to 16 of pregnancy at 1000 mg/kg bw/day. A simultaneous control group of the same size received the vehicle without test item. On Day 21 of pregnancy, the dams were killed and delivered by caesarean section. The foetuses were then examined morphologically for developmental disorders. The study showed that the repeated oral administration of the test item at a dose of 1000 mg/kg bw/day in the sensitive phase of organogenesis did not lead to any impairment of the general physical condition of the dams or impaired intrauterine development of the foetuses. The morphological examination of the foetuses with regard to stage of development, outwardly detectable anomalies as well as anomalies of the internal organs and the skeleton showed no indication of an embryotoxic or teratogenic effect of the test item. The findings observed are to be regarded as spontaneous in origin. On the basis of the results of this limit test, the NOAEL for the test item in rats following oral administration lies at 1000 mg/kg bw/day with regard to maternal and embryo-foetal toxicity and teratogenicity. No teratogenic effect was observed.

Justification for classification or non-classification

No adverse effects on reproduction toxicity observed - no classification necessary