Multi constituent substance

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study performed according to OECD guideline 414 in compliance to GLP.

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

2005

Report date:

2005

Reference 2

Reference Type:

other: Published secondary source

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Multi constituent substance

EC Number:

916-604-0

IUPAC Name:

Multi constituent substance

Details on test material:

Test substance : Basic Brown 17 (COLIPA number B007)
Batch number : 64960101
Methylsulphate anion : 11.6%
Chloride ion : 3.3%
Water : 6.5%
Basic Red 118 : 4.5%
Purity : 94.2% (HPLC)

Test animals

Species:

rat

Strain:

other: Wistar HsdBrlHan

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: distilled water

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

Animals were treated on day 5-19 of gestation

Frequency of treatment:

Animals were treated once daily

Duration of test:

Animals were sacrificed on day 20 of gestation

No. of animals per sex per dose:

Twenty five females per dose group

Control animals:

yes, concurrent vehicle

Results and discussion

Results: maternal animals

Effect levels (maternal animals)

Dose descriptor:

NOAEL

Effect level:

93 mg/kg bw/day

Based on:

act. ingr.

Basis for effect level:

other: maternal toxicity

Results (fetuses)

Effect levels (fetuses)

Dose descriptor:

NOAEL

Effect level:

186 mg/kg bw/day

Based on:

act. ingr.

Basis for effect level:

other: fetotoxicity

Fetal abnormalities

Abnormalities:

not specified

Overall developmental toxicity

Developmental effects observed:

not specified

Any other information on results incl. tables

During the study there was one unscheduled death of an animal treated at the high dose. Macroscopic examination showed enlarged adrenals, abnormal swollen intestinal tract content, dark colouration of the liver and spleen. Two females in the control group, five in the low dose group, one in the mid dose group and one in the high dose group were not pregnant at termination. Unilateral implantation was present in one female in the high dose group. The number of females with live foetuses on day 20 was 23 in the controls, 20 in the low dose group, 24 in the mid dose group and 23 in the high dose group. Scabs and hair loss were the principal clinical signs observed in the treated females during treatment period. This observation was also found to occasionanally occur in the control group. Abrasion and aggressive behaviour were noted in two different females in the high dose group on day 19 and 20 respectively. Dyspnoea was observed in one female in the low dose group on day 7. No other signs of reaction to the treatment were recorded during the daily before and after treatment observations. When compared with the controls, statistically significant reductions in body weight gain (day 9) and food consumption (day 9 and 12) were noted in the high dose group. Aligned with this observation was a statistically significant lower terminal body weight and absolute weight gain in the high dose group compared with the control. Gravid uterus weight was not affected by treatment. Post mortem examination showed the spleen was dark in colour and occasionally swollen in females of the high dose group. This was considered related to the colour of the test item. Other findings were not dose related and considered to be incidental or spontaneous. At all dose levels the average number of corpora lutea, implantations, litter sizes, live foetuses, early and late resorptions, foetal body weights, percent live male foetuses were found to be similar to the controls. No dams had litters of only resorbed conceptuses. No dead foetuses were recorded. Thirteen small foetuses were found (control 4, low dose 3, mid dose 5, high dose 1). In one mid dose foetus brain ventricles were enlarged. This was considered incidental. There were no other dose dependent, significant differences in the litter or foetal incidences of any gross external, soft tissue or skeletal alterations.

Applicant's summary and conclusion

Conclusions:

The NOAEL for maternal toxicity and fetotoxicity were considered to be 93 mg/kg bw/day and 186 mg/kg bw/day respectively.

Executive summary:

The effects of Basic Brown 17 were investigated in female rats during pregnancy and on embryo-fetal development, The test item was administered orally by gavage from day 5 through to day 19 post coitum. Three groups of 25 mated female rats were given the test item at 60, 120 and 240 mg/kg day. A control group of 25 mated female rats received the vehicle (distilled water) during the treatment period. All females were caesarean sectioned on day 20 post coitum and subjected to a post mortem examination. The number of corpora lutea, implantations, early and late intrauterine death, live and dead fetuses, uterus weight and fetal weight were reported. All fetuses were examined foe external abnormalities. Approximately one half of the fetuses in each litter were examined for fixed visceral and skeletal abnormalities. Mortality and fate of females : One high dose animal was found dead on gestation day 11. Nine females, two in the control group, five in the low dose group and one in the mid- and high dose groups were not pregnant at necropsy. In addition, one high dose female showed unilateral implantations. The number of females with live foetuses on gestation day 20 was 23 in the control, 20 in the low dose group, 24 in the mid-dose group and 23 in the high dose group. Clinical signs : No relevant clinical signs were observed in the treated females. No signs were noted at pre- and post-dose observations. Brown staining on the cage tray was noted in the high dose group. The staining observed was considered to be related to the colour of the test item, which was probably eliminated in the urine. Body weight : A statistically significant reduction in body weight gain was noted in the high dose group when compared to controls on day 9 post coitum. Food consumption : Food consumption was slightly decreased, with statistical significance, in the high dose group on days 9 and 12 post coitum when compared to the control. Terminal body weight, uterus weight and absolute weight gain : Statistically significant lower terminal body weight and consequently absolute weight gain were noted in the high dose group when compared to the control. Gravid uterus weight was not affected by treatment. Litter data and sex ratios : Litter data and sex ratios were not affected by the treatment. Macroscopic observations : A total of 14 high dose females showed abnormal colour of the spleen (dark colour). In addition, in the same group, a swollen shape of the spleen was also noted in 8 animals. External examination in foetuses : The numbers of small foetuses were comparable between groups. Visceral and skeletal examinations in foetuses : Visceral examination in foetuses did not show any dose related findings. No relevant changes that could be considered treatment related were observed at skeletal examination of foetuses. Conclusion : One high dose female was found dead on day 11 of gestation. A slight, but statistically significant decrease in food consumption was recorded in the high dose group on days 9 and 12 post coitum. Statistically significant decreases in terminal body weight and consequently absolute weight gain were observed in the high dose group when compared to the control group. Litter data and sex ratios were not affected by treatment. At macroscopic examination a total of 14 high dose females showed dark colour of the spleen. In addition, in the same group, a swollen shape of the spleen was noted in 8 animals. Visceral and skeletal examinations of treated animals did not show any dose-related findings when compared to control. On the basis of these results, the mid-dose of 120 mg/kg bw/day of the test item (93 mg/kg bw/day active) should be considered the maternal No Observed Adverse Effect Level (NOAEL). Neither embryotoxic nor teratogenic effects have been found up to the highest tested dose of 240 mg/kg bw/day (NOAEL fetotoxicity 240 mg/kg bw/day test item (186 mg/kg bw/day active)).