Tetrasodium 4-amino-5-hydroxy-6-[[2-methoxy-5-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]azo]-3-[[4-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]azo]naphthalene-2,7-disulphonate

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Two developmental toxicity studies were conducted with two structural analogues covering the respective structures/metabolites of the substance. In these studies, no adverse effects on pregnancy of foetal development have been observed.

Hence the substance Reactive Blue 250 is no expected to have any adverse effect on reproduction or foetal development.

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the study does not need to be conducted because a pre-natal developmental toxicity study is available

Reproductive effects observed:

not specified

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

Developmental/teratogenicity studies without adverse effects are available for two structural analogues which cover all functional groups and possible metabolites of Reactive Blue 250. Hence, a reproductive/developmental screening study has not to be performed according to Column 2 of REACH Annex VIII due to animal welfare reasons. Furthermore, no effects were seen on reproductive organs in the repeat dose studies in these 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 studies in conjunction with the lack of effects noted in the other toxicity studies and the similarity of the substances with respect to structure, physico-chemical, toxicokinetic and mechanistic properties are suitable to address this endpoint.

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 analogues in the pre-natal development toxicity study.

Link to relevant study records

Reference

Reference 1

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From Jan. 14, 2008 to Feb. 14, 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

other: Crl:(Wi) BR-Wistar rats

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: CHARLES RIVER/EUROPE/LABORATORIES INC., TOXI-COOP 1103 Budapest, Hungary Cserkesz utca 90.
- Age at study initiation: Males: 9-10 weeks, Females: 6-8 weeks
- Weight at study initiation: Virgin female rats of comparable weight were used for mating. The weight variation was 198-285 g on the first day (Day 0) of gestation.
- Housing: Type II and III polypropylene/polycarbonate cages, with stainless steel covers equipped by self-feeding basket
- Bedding: laboratory bedding material, changed twice a week
- Diet: Ssniff SM R/M-Z+H "Autoclavable complete feed for rats and mice – breeding and maintenance", ad libitum
- Water: tap water, as for human consumption, ad libitum
- Acclimation period: 5 d

ENVIRONMENTAL CONDITIONS
- Temperature: 22±3°C
- Humidity: 30-70%
- Photoperiod: 12 h light/dark cycle

IN-LIFE DATES: From: Jan. 14, 2008 To: Feb. 14, 2008

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

distilled

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: The test substance was suitable for oral (by gavage) administration when suspended in distilled water. The test substance was administered at appropriate concentrations prepared with the vehicle. The formulations of each concentration were used for the treatment within 60 minutes after preparation.

VEHICLE: distilled water

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Sampling for analytical control of dosing was made on the second and last week of the treatment period by the Analytical Laboratory of LAB Research Ltd. The analysis was carried out using HPLC. All the formulations proved to be homogeneous. Measured concentrations varied between 99 and 104 % of the nominal concentrations.

Details on mating procedure:

The oestrus cycle of female animals were examined before pairing. After acclimatisation, the females were paired according to their oestrus cycle to males in the morning for 3 h (1 male: 1 to 3 females) until the number of sperm positive females / group achieved 22. Vaginal smears were prepared from each female, stained with 1% aqueous methylene blue solution and examined for presence of sperm. The day of mating was regarded as Day 0 of pregnancy (vaginal plug and/or sperm in the vaginal smear). Sperm positive females were separated and caged in groups of 2 to 4 animals.

Duration of treatment / exposure:

The test substance was administered to the sperm positive females from Day 5 up to and including the Day 19 of gestation.

Frequency of treatment:

Once a day

Duration of test:

In life phase of the test was concluded by the cervical dislocation of all sperm positive females on Day 20 of gestation

No. of animals per sex per dose:

22 sperm-positive female/group

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: The dose levels for the study were selected on the basis of a dose range finding study, in which groups of 5 sperm positive female rats were administered 62.5, 100 and 1,000 mg/kg bw/day by oral gavage from Day 5 to Day 19 post coitum. The 1,000 mg/kg bw/day dose of the test substance was slightly toxic to the dams as it led to slightly reduced body weight and body weight gain, while no effect on the intrauterine development of the embryos and foetuses at any of the doses was seen. Hence the 1,000 mg/kg bw/day was considered suitable for the top dose in the main teratology study. (Study code: 07/583-105PE)
- Rationale for animal assignment: The sperm positive females were allocated to each experimental group on each mating day in such a way that the group averages of the body weight were as similar as possible on the Day 0 of pregnancy. Females paired with the same male were allocated to different groups on the same mating day.

Maternal examinations:

Clinical Observations:
A careful clinical observation was made after dosing at least once a day and a cage side clinical observation was made in the afternoon. Individual observation included the check of behaviour and general condition. Checking for death was made at least once daily.

Body Weight:
The body weight of the male animals was not measured.
The body weight of the female rats was measured at least once in the pre-mating period, but was not statistically evaluated. Body weight of sperm positive females was measured on gestation Days 0, 3, 5, 8, 11, 14, 17 and 20 (accuracy of 1 g). Corrected body weight was calculated on the 20th day of pregnancy (body weight on Day 20 minus the weight of the gravid uterus).

Food Consumption:
The food consumption was measured between gestation Days 0 to 3, 3 to 5, 5 to 8, 8 to 11, 11 to 14, 14 to 17 and 17 to 20 by re-weighing the non-consumed diet (accuracy: 1 g).

Examination for Sign of Implantation:
On gestation Days 13 and/or 14 the sperm positive females were checked for the presence of vaginal bleeding which indicates the implantation of conceptuses.

Necropsy:
All sperm positive females were sacrificed by CO2 gas anaesthesia followed by cervical dislocation on Day 20 of gestation. The abdomen was opened, the uterus with cervix and the left ovary were removed and weighed. The right ovary was placed into a petri dish after removal. After removing the uterus and its content gross pathology of dams' viscera was performed. There were no organs and tissues with undiagnosed macroscopic findings examined histologically.
The number of corpora lutea in each ovary and implantation sites in each uterine horn, the number of live foetuses, early and late embryonic death and foetal death were counted. Animals with unambiguous implantation sites but no foetuses were found, were considered as pregnant. Foetuses were removed from the opened uterus, viability assessments were performed and after cutting the umbilical cord euthanasia of the viable foetuses was made by subcutaneous injection of 0.01 mL Euthasol in the femur region. Each live foetus was weighed individually (accuracy 0.01 g), and was subjected to external examination. The placentas were examined externally. The gender of foetuses was determined according to the anogenital distance.

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No data
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes

Fetal examinations:

- Body weight: Yes: all per litter
- External examinations: Yes: all per litter
- Visceral examinations (including head): Yes: half per litter
- Skeletal examinations: Yes: half per litter
-

Statistics:

The homogeneity of variance between groups was checked by Bartlett’s homogeneity of variance test.
Where no significant heterogeneity was detected a one-way analysis of variance (ANOVA) was carried out. If the obtained result was significant Duncan Multiple Range test was used to access the significance of inter-group differences. Getting significant result at Bartlett’s test the Kruskal-Wallis analysis of variance was used and the inter-group comparisons were performed using Mann-Whitney U-test.

A foetus was considered as retarded in body weight, when its weight was below the average minus twofold standard deviation of all control foetuses.

Clinical signs:

no effects observed

Mortality:

no mortality observed

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:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not examined

Behaviour (functional findings):

no effects observed

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

At necropsy, findings of slightly pale areas on the liver was recorded for the dams with a statistical significance in the 1000 mg/kg bw/day dose group.

Neuropathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Number of abortions:

no effects observed

Pre- and post-implantation loss:

no effects observed

Total litter losses by resorption:

no effects observed

Early or late resorptions:

no effects observed

Dead fetuses:

no effects observed

Changes in pregnancy duration:

no effects observed

Changes in number of pregnant:

no effects observed

Details on maternal toxic effects:

At necropsy, findings of slightly pale areas on the liver was recorded for the dams with a statistical significance in the 1000 mg/kg bw/day dose group.

Dose descriptor:

NOEL

Effect level:

250 mg/kg bw/day (actual dose received)

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

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

Key result

Abnormalities:

no effects observed

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

Changes in postnatal survival:

no effects observed

External malformations:

effects observed, non-treatment-related

Skeletal malformations:

effects observed, non-treatment-related

Visceral malformations:

no effects observed

Other effects:

no effects observed

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects: no effects

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

Key result

Abnormalities:

no effects observed

Key result

Developmental effects observed:

no

Details on maternal observations:

There was no maternal mortality or maternal clinical signs in any group. The test substance had no effect on the body weight, body weight gain, gravid uterine weight, corrected body weight, corrected body weight gain or food consumption of dams.

At necropsy, findings of slightly pale areas on the liver was recorded for the dams with a statistical significance in the 1000 mg/kg dose group.

 

Details on foetal observations:

The intrauterine mortality and sex distribution of foetuses were unaffected by the treatment. There was no difference between groups in the number of corpora lutea or implantations. The body weight of foetuses was similar in all experimental groups. Reddish discolouration of the amniotic epithelium was recorded for the majority of the foetuses in the 1000 mg/kg dose group that suggested that the coloured test substance (or a coloured metabolite) crossed the placenta. Oedematous placentas were found only in the 1000 mg/kg bw/day dose group, this was attributed to the treatment with the test substance. Three multiple malformed foetuses were found at external examination in the 1000 mg/kg bw/day dose group. Two of these foetuses were found in one single litter and had visceral multiple malformations. All of the three foetuses had severe skeletal malformations. The malformations were of very low incidence and taking into account the findings from the whole study and historic data, were considered to reflect spontaneous malformations and are not likely to be related to test substance.

Conclusions:

The substance was found to have no developmental toxicity / teratogenic effects. The NOAEL for maternal toxicity and developmental toxicity / teratogenic effects was determined to be 1000 mg/kg bw/day.

Executive summary:

A study was conducted to assess the developmental toxicity of the test substance according to OECD Guideline 414, in compliance with GLP.

Groups of 22 sperm-positive female rats were treated orally in one control group and three dose levels of 62.5, 250 and 1000 mg/kg bw/day (5 mL/kg) from Day 5 to Day 19 post-coitum. Rats were examined daily for morbidity and clinical signs. Body weight was recorded on Days 0, 3, 5, 8, 11, 14, 17 and 20 of gestation. Food consumption was determined on Days 0-3, 3-5, 5-8, 8-11, 11-14, 14-17 and 17-20 of gestation. Caesarean section and gross-pathology were performed on Day 20 of pregnancy. Implantations, early and late resorptions, live and dead foetuses in each uterine horn and the number of corpora lutea in each ovary were recorded. Each foetus was weighed and examined for sex and gross external abnormalities. The placentas were examined externally. About half of each litter was subjected to visceral examination and the other half to skeletal examination (with double staining). All abnormalities found during the foetal examinations were recorded.

No maternal mortality or maternal clinical signs were recorded in any group. The test substance had no effect on the body weight, body weight gain, gravid uterine weight, corrected body weight, corrected body weight gain or food consumption of dams. At necropsy, slightly pale areas on the liver were recorded for the dams with a statistical significance in the 1000 mg/kg bw/day dose group. The intrauterine mortality and sex distribution of foetuses were unaffected by the treatment. There was no difference between groups in the number of corpora lutea or implantations. The body weight of foetuses was similar in all experimental groups. Reddish discolouration of the amniotic epithelium was recorded for the majority of the foetuses in the 1,000 mg/kg dose group, suggesting that the test substance (or a coloured metabolite) crossed the placenta. Oedematous placentas were found only in the 1000 mg/kg bw/day dose group. This was attributed to treatment with the test substance. Three multiple malformed foetuses were found at external examination in the 1000 mg/kg bw/day dose group. Two of these foetuses were found in one single litter and had visceral multiple malformations. All of the three foetuses had severe skeletal malformations. The malformations were of very low incidence. Taking into account the findings from the whole study and historic data, they were considered to reflect spontaneous malformations rather than related to test substance exposure.

Under the study conditions, the substance was found to have no developmental toxicity / teratogenic effects. The NOAEL for maternal toxicity and developmental toxicity / teratogenic effects wasdetermined to be 1000 mg/kg bw/day.

Reference 2

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

14. Sep to 25 Nov 1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

GLP compliance:

yes

Limit test:

yes

Species:

rat

Strain:

Wistar

Details on test animals or test system 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

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

Clinical signs:

no effects observed

Mortality:

no mortality observed

Food efficiency:

no effects observed

Gross pathological findings:

no effects observed

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.

Remarks on result:

not determinable due to absence of adverse toxic effects

Dose descriptor:

NOEL

Effect level:

>= 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

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

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

Abnormalities:

no effects observed

Developmental effects observed:

no

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.

 

A study was conducted to assess the developmental toxicity of the test substance (structural analogue 03) according to OECD Guideline 414, in compliance with GLP. 

Groups of 22 sperm-positive female rats were treated orally in one control group and three dose levels of 62.5, 250 and 1000 mg/kg bw/day (5 mL/kg) from Day 5 to Day 19 post-coitum. Rats were examined daily for morbidity and clinical signs. Body weight was recorded on Days 0, 3, 5, 8, 11, 14, 17 and 20 of gestation. Food consumption was determined on Days 0-3, 3-5, 5-8, 8-11, 11-14, 14-17 and 17-20 of gestation. Caesarean section and gross-pathology were performed on Day 20 of pregnancy. Implantations, early and late resorptions, live and dead foetuses in each uterine horn and the number of corpora lutea in each ovary were recorded. Each foetus was weighed and examined for sex and gross external abnormalities. The placentas were examined externally. About half of each litter was subjected to visceral examination and the other half to skeletal examination (with double staining). All abnormalities found during the foetal examinations were recorded.

No maternal mortality or maternal clinical signs were recorded in any group. The test substance had no effect on the body weight, body weight gain, gravid uterine weight, corrected body weight, corrected body weight gain or food consumption of dams. At necropsy, slightly pale areas on the liver were recorded for the dams with a statistical significance in the 1000 mg/kg bw/day dose group. The intrauterine mortality and sex distribution of foetuses were unaffected by the treatment. There was no difference between groups in the number of corpora lutea or implantations. The body weight of foetuses was similar in all experimental groups. Reddish discolouration of the amniotic epithelium was recorded for the majority of the foetuses in the 1,000 mg/kg dose group, suggesting that the test substance (or a coloured metabolite) crossed the placenta. Oedematous placentas were found only in the 1000 mg/kg bw/day dose group. This was attributed to treatment with the test substance. Three multiple malformed foetuses were found at external examination in the 1000 mg/kg bw/day dose group. Two of these foetuses were found in one single litter and had visceral multiple malformations. All of the three foetuses had severe skeletal malformations. The malformations were of very low incidence. Taking into account the findings from the whole study and historic data, they were considered to reflect spontaneous malformations rather than related to test substance exposure.

Under the study conditions, the substance was found to have no developmental toxicity / teratogenic effects. The NOAEL for maternal toxicity and developmental toxicity / teratogenic effects was determined to be 1000 mg/kg bw/day.

Justification for classification or non-classification

No adverse effects on reproduction toxicity observed - no classification necessary