Benzenamine, N,N-dimethyl-, oxidized, molybdatetungstatephosphates

Endpoint summary

Description of key information

OECD422, oral, rat (m/f), NOAEL = 1000 mg/kg bw /day

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

repeated dose toxicity: oral, other

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29 Aug 2017 - 22 Feb 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 10 weeks
- Weight at study initiation: Males: 331-431 g Females: 179-245 g
- Housing: Cages with standard, granulated, S8-15 sawdust bedding
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 30 - 70
- Air changes (per hr): 15-20
- Photoperiod (hrs dark / hrs light): 12/ 12

Route of administration:

oral: gavage

Details on route of administration:

Oral, by gastric gavage. For each dose group the order of administration was all males first and then all females. Each day of treatment the starting order was alternated between males and females.

Vehicle:

arachis oil

Details on oral exposure:

- VEHICLE
- Batch number KMO9422, KM09047
- Expiry date: August 2018
- Storage conditions: Stored at ambient conditions
- Safety precautions: Routine hygienic procedures (nitrile gloves, goggles, face mask)

- Concentration in vehicle: 0, 100, 300 and 1000 mg/kg/day
- Administration volume: 5 mL/kg

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Validation done at 20-200 mg/mL demonstrates that formulations were stable for 8 days at room temperature (20 ± 5 ºC) and in the dark.

The results showed that analyzed formulations were within 19% of their nominal concentration. The coefficient of variation (CV) was not more than 2.13%. Preparation of all formulations was considered correct, taking into account the acceptance ranges.

The analysis of control group formulations showed that no contamination was present.

Duration of treatment / exposure:

5 - 8 weeks

Dose / conc.:

0 mg/kg bw/day (nominal)

Dose / conc.:

100 mg/kg bw/day (nominal)

Dose / conc.:

300 mg/kg bw/day (nominal)

Dose / conc.:

1 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

3 groups of 12 females
3 gouprs of 12 males
1 control group of 12 females
1 control group of 12 males

Control animals:

yes

Observations and examinations performed and frequency:

VIABILITY/ MORTALITY/ CAGE-SIDE OBSERVATIONS
Visually inspected twice daily for evidence of reaction to treatment or ill-health.

BODY WEIGHT:
- Time schedule for examinations: On day 1 and weekly thereafter

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:

no effects observed

Water consumption and compound intake (if drinking water study):

no effects observed

Ophthalmological findings:

not specified

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not specified

Behaviour (functional findings):

no effects observed

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

The changes observed in the brain weights in males and females and in liver weights in males at 1000 mg/kg/day when compared to control were considered incidental taking into account the magnitude of the change, the lack of a dose-effect relationship and that there was no correlation with clinical pathology or histopathology.

Gross pathological findings:

no effects observed

Neuropathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Other effects:

not specified

Key result

Dose descriptor:

NOEL

Effect level:

ca. 1 000 mg/kg diet

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

clinical signs

food consumption and compound intake

other: clinical pathology, organ weights or histopathology

Key result

Critical effects observed:

no

Conclusions:

The No Observed Effect Level (NOEL) for systemic toxicity was considered to be 1000 mg/kg/day, taking into account that there was no effect on body weight, food consumption, clinical signs, clinical pathology, organ weights or histopathology.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

GLP guideline study

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

In a combined repeated dose oral gavage toxicity study with the reproduction/developmental toxicity screening (OECD 422), three groups of 12 male and 12 female rats received Pigment Violet 3 PTM at the doses of 100, 300 and 1000 mg/kg/day, respectively. Males were treated continuously for two weeks before pairing up to necropsy, after a minimum of 32 consecutive days. Females were treated continuously for two weeks before pairing, throughout pairing and gestation, and until Day 14-16 of lactation (the day before sacrifice). Females were allowed to litter and rear their offspring, and litters were killed on Day 13-15 of lactation (the day before the corresponding female was killed). F1 generation received no direct administration of the test item; any exposure was in utero or via the milk. A similarly constituted control group received the vehicle, arachis oil.

During the study, mortality, clinical signs, sensory reactivity observations, grip strength, motor activity, body weight, food consumption, hematology, coagulation, blood biochemistry, pre-coital interval, mating performance, fertility, gestation length, organ weight and macroscopic examination were evaluated. Clinical signs, behavior assessment, litter size, survival, sex ratio, body weight and macropathology were also assessed for all offspring.

The study results can be summarized as follows:

• There were no deaths considered related to treatment with Pigment Violet 3 PTM.
• Administration of Pigment Violet 3 PTM at 100, 300 or 1000 mg/kg/day was considered not to have any effects on clinical condition, body weight, food consumption, motor activity, sensory reactivity and grip strength, pre-coital interval, mating performance and fertility, or gestation length. Violet feces due to the test item color were recorded during treatment in all test-item-administered groups. Salivation was recorded in all test-item-administered groups; the incidence was dose-related in both sexes and is most likely due to taste aversion (from gavage dosing).
• All females allocated to the study showed regular 4-day estrous cycles before and during treatment. At termination, all reproductive phase females showed diestrus with the exception of 2/12 animals at 100 mg/kg/day, which had recovered estrous cycle.
• There were no differences in hematology, coagulation, clinical biochemistry or organ weights considered treatment-related. T4 analyses of samples in Main study males and F1 offspring on day 13 did not reveal any differences that could be attributable to treatment.
• There were no macroscopic findings that could be considered test-item-related.
• Histopathology reveals that there were no treatment-related effects in any of the examined organs or in the reproductive organs or mammary glands.
• There was no effect on offspring survival due to Pigment Violet 3 PTM or on litter size, sex ratio, clinical signs, body weights, ano-genital distances or macropathology.

In conclusion, the effects of oral (gavage) administration of Pigment Violet 3 PTM to Wistar rats receiving 100, 300 or 1000 mg/kg/day for 14 days prior to mating and until sacrifice can be summarized as follows:

Systemic toxicity:

The No Observed Effect Level (NOEL) for systemic toxicity was considered to be 1000 mg/kg/day, taking into account that there was no effect on body weight, food consumption, clinical signs, clinical pathology, organ weights or histopathology.

Reproductive / developmental toxicity:

The No Observed Effect Level (NOEL) for reproductive / developmental toxicity was considered to be 1000 mg/kg/day, taking into account that there was no effect on estrous cycle, pre-coital interval, mating performance, fertility and gestation length or in the offspring on litter size, survival, sex ratio, clinical signs, body weights, ano-genital distances or macropathology.

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008.

Based on available data on repeated dose toxicity, the substance is not classified for specific target organ toxicity following repeated exposure according to Regulation (EC) No 1272/2008 (CLP), as amended for the tenth time in Regulation (EU) No 2017/776.