3-[4-[[5-(1,1-dimethylpropyl)-2-hydroxy-3-nitrophenyl]azo]-4,5-dihydro-3-methyl-5-oxo-1H-pyrazol-1-yl]benzenesulphonamide, reaction products with aqueous organic chromium(III) complex sodium salt, then laked with acidified C10-14-tert-alkyl(linear and branched) amines

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2014-08-25 to 2016-04-12

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:

2016

Report date:

2016

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: OECD guideline 421 (July 1995)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: OPPTS 870.3550 (July 2000)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

3-[4-[[5-(1,1-dimethylpropyl)-2-hydroxy-3-nitrophenyl]azo]-4,5-dihydro-3-methyl-5-oxo-1H-pyrazol-1-yl]benzenesulphonamide, reaction products with aqueous organic chromium(III) complex sodium salt, then laked with acidified C10-14-tert-alkyl(linear and branched) amines

EC Number:

939-383-2

Molecular formula:

UVCB substance

IUPAC Name:

3-[4-[[5-(1,1-dimethylpropyl)-2-hydroxy-3-nitrophenyl]azo]-4,5-dihydro-3-methyl-5-oxo-1H-pyrazol-1-yl]benzenesulphonamide, reaction products with aqueous organic chromium(III) complex sodium salt, then laked with acidified C10-14-tert-alkyl(linear and branched) amines

Test material form:

solid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 002-122106

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature

Test animals

Species:

rat

Strain:

other: Wistar rats, strain Crl:WI(Han)

Details on test animals or test system and environmental conditions:

TEST ANIMALS:
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH
- Age at study initiation: 11-13 weeks
- Weight at study initiation: 321.1 g – 355.1 g (males); 196.3 g – 222.2 g (females)
- Fasting period before study: No
- Housing: During the study period, the rats were housed individually in Polycarbonate cages type III supplied by Becker & Co., Castrop-Rauxel, Germany (floor area of about 800 cm²) and TECHNIPLAST, Hohenpeißenberg, Germany, with the following exceptions: During overnight matings, male and female mating partners were housed together in Polycarbonate cages type III. Pregnant animals and their litters were housed together until PND 4.
- Diet: Ground Kliba maintenance diet mouse/rat “GLP” meal, supplied by Provimi Kliba SA, Kaiseraugst, Switzerland; ad libitum
- Water: ad libitum
- Acclimation period: About 5 days

ENVIRONMENTAL CONDITIONS:
- Temperature: 20-24 °C
- Humidity: 30-70 %
- Air change: 15 times per hour
- Photoperiod:12 hours light (from 6.00 h to 18.00 h) and 12 hours darkness (from 18.00 h to 6.00 h)

Administration / exposure

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
The required quantity of test substance was weighed in a beaker depending on the test group and thoroughly mixed with a small amount of food (Ground Kliba maintenance diet mouse/rat). Then further amounts of food were added to this premix and thoroughly mixed. Afterwards, further amounts of food, depending on the dose group, were added to this premix in order to obtain the desired concentrations. Mixing of this final mix was carried out for about 3 minutes in a laboratory mixer.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Analytical verifications of the stability of the test substance in the diet for a period of 35 days at room temperature were carried out before the study was initiated with a comparable batch. Samples of the test substance preparations were sent to the analytical laboratory once during the study period for verification of the concentration. The samples, which were taken for the concentration control analysis at the beginning of the administration period, were also used to verify the homogeneity.

Details on mating procedure:

- M/F ratio per cage: 1:1
- Length of cohabitation: Maximum of 2 weeks
- Proof of pregnancy: Proof of pregnancy: Detection of mating was confirmed by sperm in vaginal smear. The day on which sperm were detected was denoted "GD 0" and the following day "GD 1".

Duration of treatment / exposure:

30 days (males);
50 – 56 days (females)

Frequency of treatment:

Daily

Duration of test:

30 days (males);
50 – 56 days (females)

No. of animals per sex per dose:

10

Control animals:

other: yes, treated with diet only

Examinations

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: At least once daily
- Cage side observations included morbidity, pertinent behavioral changes and signs of overt toxicity.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily

BODY WEIGHT: Yes
- Time schedule for examinations: Once a week. The follwoing exceptions are notable for the female animals: During the mating period the parental females were weighed on the day of positive evidence of sperm (GD 0) and on GD 7, 14, and 20. Females with litter were weighed on the day after parturition (PND 1) and on PND 4.

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

Ovaries and uterine content:

not examined

Fetal examinations:

not examined

Statistics:

Statistical analyses were performed with the following statistical tests:
Food consumption, body weight and body change, gestation days: DUNNET test (two-sided)
Male and female mating indices, male and female fertility indices, females mated, females delivering, gestation index, females with liveborn pups, females with all stillborn pups: FISHER'S EXACT test (one-sided)
Mating days until day 0 pc, % postimplantation loss, pups stillborn, % perinatal loss, implantation sites, pups delivered, pups liveborn, live pups day x, viability index: WILCOXON test (one-sided) with BONFERRONI-HOLM
% live male day x, % live female day x: WILCOXON test (two-sided)
Weight parameters (pathology): KRUSKAL-WALLIS test (two-sided) and WILCOXON-test (two-sided)

Indices:

Viability index (%), Sex ratio (%), stillborn and liveborn pups (%), postimplantation loss (%), perinatal loss (%)

Results and discussion

Results: maternal animals

Maternal developmental toxicity

Details on maternal toxic effects:

Maternal toxic effects: yes

Details on maternal toxic effects:
Test following substance-related adverse effects/findings were noted at 812 mg/kg bw/d (12000 ppm):
- Decreased food consumption in the females during premating days 0-7 and 0-13 (about 14% and 9% below control, respectively), during the entire gestation period (up to 15% below control) and during the entire lactation period (about 31% below control)
- Decreased body weights in the females on GD 20 (about 7% below control) and during the entire lactation period (up to 14% below control)
- Decreased body weight change in the females during PND 1-4 (about 67% below control)
- Reduced terminal body weight in females (94%)

Effect levels (maternal animals)

1

Key result

Dose descriptor:

NOAEL

Effect level:

261 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: developmental toxicity

2

Key result

Dose descriptor:

NOAEL

Effect level:

261 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

Results (fetuses)

Details on embryotoxic / teratogenic effects:

Test following substance-related adverse effects/findings were noted at 812 mg/kg bw/d (12000 ppm):
- Decreased pup body weights on PND 4 (about 25% below control)
- Decreased pup body weight gain on PND 1 - 4 (about 49% below control)

Effect levels (fetuses)

Key result

Dose descriptor:

LOAEL

Effect level:

812 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Decreased pup body weights on PND 4 and decreased pup body weight gain on PND 1 - 4

Fetal abnormalities

Abnormalities:

not specified

Overall developmental toxicity

Developmental effects observed:

not specified