2,4-bis(((2-(dimethylammonio)ethyloxy)carbonyl)phen-2-ylazo)benzene-1,3-diolbis(methanesulfonate)

Toxicity to reproduction

Administrative data

Endpoint:

one-generation reproductive toxicity

Remarks:

based on test type (migrated information)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD Guideline study 415

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2000

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 415 [One-Generation Reproduction Toxicity Study (before 9 October 2017)]

GLP compliance:

yes

Test material

Reference

Name:

Unnamed

Type:

Constituent

Details on test material:

- Name of test material (as cited in study report): Basisch Gelb 8511
- Physical state: powder
- Analytical purity: 98,9%
- Lot/batch No.: ZD 00969/086

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Boehringer Ingelheim Pharma KG
- Age at study initiation: 37 +- 1 day at the beginning of treatment
- Weight at study initiation: Males: 148.0 - 184.8 g (mean 168.0 g); Females: 116.7 - 155.9 g (mean 138.0g);
- Housing: DK III stainless steel wire mesh cages, Becker A& Co.; floor area of about 800 cm²
- Dieta: ad libitum
- Water: ad libitum
- Acclimation period: 9 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24°C
- Humidity (%): 30 - 70 %
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

olive oil

Details on exposure:

As the test substance is sensitive to hydrolysis, it was administered as a suspension in olive oil (EAB, 1997; pharmaceutical). To prepare the suspension, the appropriate amount of test substance was weighed, depending on the dose group. Then the olive oil was filled up to the desired volume and mixed using an Ultra Turrax. During the administration the test substance preparations were kept homogenous with a magnetic stirrer. In respect to the proven stability the test substance preparations were prepared twice a week before dosing.

Details on mating procedure:

- M/F ratio per cage:1:1
- Length of cohabitation: overnight for a maximun of 3 weeks
- Proof of pregnancy: sperm in vaginal smear] referred to as day 0 of pregnancy
- Any other deviations from standard protocol: exception in few matings a ration of 1:2 (males:females)due to premature mortalities of single males.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The stability of the substance in olive oil was demonstrated for a period of 4 days at room temperature before the start of the study. Homogeneity analyses were performed at the start of the administration period in samples of the high and low concentration drawn from bottom, middle and top of the substance preparations. These samples also served for concentration control analyses for these groups. Additional concentration control analyses were performed at the start of the administration period with samples from the mid concentration drawn from the middle of the substance preparation, at the middle and towards the end of the administration period with samples from all concentrations.

Duration of treatment / exposure:

Premating exposure period: min. 74 days

Frequency of treatment:

7 days per week

Doses / concentrations

Remarks:

Doses / Concentrations:
2, 10, 60 mg/kg body weight/day
Basis:

No. of animals per sex per dose:

25

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale:
In a previous subchronic toxicity study Basisch Gelb 8511 caused a range of effects including mortalities at the top dose of 180/120 mg/kg body weight. Clinical symptoms were noted down to the lowest dose of 10 mg/kg body weight. However, at 10mg/kg body weight only one female showed respiratory sounds. Therefore, the following doses were selected:
2 mg/kg: as the expected "no observed adverse effect level"
10 mg/kg: as intermediate dose level
60 mg/kg: as highest dose level

The oral route was selected since this has proven to be suitable for the detection of a toxicological hazard.

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: Yes
-Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: once a week at the same time of the day

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined:F0 and F1: once a week (in general for a period of 7 days). After the 10th weed, food consumption of the females during pregnancy was determined weekly on days 0, 7, 14, 20 p.c.; during the lactation period food consumption was determined on days 1, 4, 7 and 14 p.p.

Results and discussion

Results: P0 (first parental generation)

Effect levels (P0)

Dose descriptor:

NOAEL

Effect level:

10 mg/kg bw/day

Sex:

male/female

Results: F1 generation

Effect levels (F1)

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

60 mg/kg bw/day

Sex:

male/female

Overall reproductive toxicity

Reproductive effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

Under the conditions of this study BASISCH GELB 8511 had no adverse effects on reproductive performance or fertility of the F0 parental animals of all substance-treated groups (2, 10 and 60 mg/kg body weight/day). Mating behavior, conception, gestation, parturition, lactation and weaning as well as gross findings were similar between the substance-treated rats and the corresponding controls.

General effects in the F0 parental animals were limited to abnormal clinical findings. Respiratory sounds, discolored urine and a single case of fur smeared with urine were seen at 60 mg/kg body weight/day. The discoloration of the urine was related to the physical properties of the test compound (dyestuff) indicative for systemic availability.
The affection of the respiratory system was probably due to an unintentional aspiration of the test substance as a consequence of attempts to vomit the test substance.

Substance induced signs of developmental toxicity were not observed in progeny of the F0 parents at any dose level.

Therefore, under the conditions of this study the NOAEL (no observed adverse effect level) for reproductive performance and fertility is 60 mg/kg body weight/day for the F0 parental rats.

The NOAEL for general effects of the test substance is 10 mg/kg body weight/day for the F0 parental males and females.

The NOAEL for developmental toxicity (growth and development of the offspring) could be fixed at 60 mg/kg body weight/day for the F1 progeny.

Thus, indications for impaired reproductive performance and fertility or for developmental toxicity were not observed up to the high dose of 60 mg/kg body weight/day.

Executive summary:

BASISCH GELB 8511 was administered as an oily suspension to Wistar rats over one parental (F0) generation by stomach tube at dosages of 2; 10 or 60 mg/kg body weight/day.

There were no indications from the clinical and pathological examinations, that the administration had adverse effects on reproductive performance or fertility of the F0 parental animals of all substance-treated groups. Mating behavior, conception, gestation, parturition, lactation and weaning as well as gross pathology were similar between the substance-treated rats and the corresponding controls. Most of the F0 parental rats proved to be fertile within the scheduled mating interval. Only few male and female F0 parental rats had to be reevalua­ted for fertility. With exception of one high dose female fertility was confirmed for all of these animals. According to the results of pathology no relevant gross lesions that may explain the infertility of this female was noted. Therefore, with respect of the results of scheduled mating and reevaluation of fertility the overall conclusion was drawn that there were no substance-related effects on the fertility of male and female F0 parental animals.

General effects in the F0 parental animals were limited to abnormal clinical findings. Salivation and retching, both shortly after treatment, were noted in all dose groups (more pronounced in males than in females), whereas respiratory sounds, discolored urine and fur smeared with urine were exclusively seen in male and female F0 parental rats of the high dose group. However, although salivation and retching (attempts to vomit the test substance) shortly after treatment were associated to the mode of administration by gavage, these findings were regarded to represent not a toxicologically relevant effect. The discoloration of the urine was, however, related to the physical properties of the test compound (dyestuff) indicative for systemic availability but was not assessed as an adverse effect per se. The respiratory sounds were probably due to an unintentional aspiration of the test substance as a consequence of attempts to vomit as it was noted in the previous subchronic toxicity study also (BASF 1999). Moreover, in that study a histological correlate was found. Consequently, the respiratory sounds in this study were also assessed as indication of an affection of the respiratory system.

The premature deaths of one male rat of the mid dose group and of six males of the high dose group were assessed to be not substance-related, but they are the consequence of misgavage, probably due to the strong defensive reaction noted in most of the animals of the high dose group.

The gross pathological evaluation yielded no indications for substance-induced general, systemic toxicity in the F0 parental animals in none of the male and female F0 parental animals of any group.

Substance-induced signs of developmental toxicity were not observed in progeny of the F0 parents at any dose level.

Therefore,under the conditions of this study the NOAEL (no observed adverse effect level) for reproductive performance and fertility is 60 mg/kg body weight/day for the F0 parental rats.

The NOAEL for general effects of the test substance is 10 mg/kg body weight/day for the F0 parental males and females.

The NOAEL for developmental toxicity (growth and development of the offspring) could be fixed at 60 mg/kg body weight/day for the F1 progeny.

Thus, indications for impaired reproductive performance and fertility or for developmental toxicity were not observed up to the high dose of 60 mg/kg body weight/day.