Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdatephosphates

Endpoint summary

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29 August 2017 - 22 February 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:

not specified

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Specific details on test material used for the study:

Batch number: 982432
Purity: Preparation containing ≥80% UVCB (treat as 100%)
Storage conditions: Stored at ambient conditions in the dark (away from direct light)

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, France
- Females (if applicable) nulliparous and non-pregnant: no
- Age at study initiation: 10 weeks both male and female
- Weight at study initiation: Males: 331-431 g, Females: 179-245 g
- Housing: Cages with standard, granulated, S8-15 sawdust bedding (J. Rettenmaier & Söhne)
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: Five days after arrival and before the start of pre-test. After acclimatization period, the animals were subjected to a 15-day pre-test period.

DETAILS OF FOOD AND WATER QUALITY: Pelleted standard Teklad 2014C and 2914C rat/mouse maintenance diet ad libitum (supplied by Envigo RMS, S.L.)..
Pelleted standard Teklad 2018C rat/mouse maintenance diet (supplied by Envigo RMS, S.L.) ad libitum, for lactating females and pups (until sacrifice).

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24 ºC
- Humidity (%): 30 and 70%
- Air changes (per hr): 15-20 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours fluorescent light/12 hours dark.

Route of administration:

oral: gavage

Details on route of administration:

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.
Formulations were maintained under agitation between 5 to 24 hours (from start of agitation until the end of administration).

Vehicle:

arachis oil

Details on oral exposure:

- PREPARATION OF DOSING SOLUTIONS: The required amount of test item was weighed in a disposable container. Approximately 80% of the final volume requested of arachis oil was poured in another disposable container. The test item was transferred to the container with the arachis oil and the mixture poured into an ultra turrax or a homogenizer to achieve a homogeneous suspension. The suspension was
poured into a volumetric flask or test tube. The disposable container that contained the mixture with the remaining amount of vehicle was washed and the contents added to the flask or test tube to make up to the mark. An adjustment volume was first added to the volumetric container and then to the container containing the formulation to reach the final requested volume of the formulation. Finally, the suspension was submitted to magnetic stirring for 5-10 minutes before sampling for dilution.

- VEHICLE
- Lot/batch no. (if required): KMO9422, KM09047
- Purity: not specified

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

12-mL aliquots (in duplicate) were taken from each formulation to be analyzed. Each aliquot was taken from the formulation freshly prepared and poured into an amber glass vial. The second aliquot was considered as a contingency sample.

Duration of treatment / exposure:

5-8 weeks

Frequency of treatment:

Once daily
- F0 males: Two weeks prior to mating start until the day before sacrifice (for five weeks of dosing). They were then killed.
- F0 females: Two weeks prior to mating start until day 13/15 of lactation, including the day before sacrifice.
- F1: Potential indirect exposure in utero and through the milk during lactation

Dose / conc.:

0 mg/kg bw/day (nominal)

Remarks:

Control (Group 1)

Dose / conc.:

100 mg/kg bw/day (nominal)

Remarks:

Group 2

Dose / conc.:

300 mg/kg bw/day (nominal)

Remarks:

Group 3

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Remarks:

Group 4

No. of animals per sex per dose:

12

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: It was considered a suitable dose level range, based on the preliminary results obtained in the previous non-GLP study HJ88HL 14-day Oral (Gavage) Dose-Range Toxicity Study for OECD 422 conducted at Envigo CRS, S.A.U.
- The high dose was selected as no toxicity was observed in the preliminary study at 1000 mg/kg/day and considering it as a limit dose to be tested.
- Intermediate and low dose levels were selected considering approximately a 3-fold interval between doses.

- Rationale for animal assignment (if not random): Random

Positive control:

none

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once a week

BODY WEIGHT: Yes
- Time schedule for examinations:
- pre-test: once a week
- treatment: On day 1 and weekly thereafter
- mating: day 1
- post-mating: weekly
- gestation: On days: 0, 7, 14, 20
- Lactation: On days: 1, 4, 9 and 13

FOOD CONSUMPTION:
- Food consumption: Yes weekly

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at termination
- Anaesthetic used for blood collection: Yes Isoflurane
- Animals fasted: Yes
- How many animals: Males:
- Group 1: 1, 2, 3, 4 and 5
- Group 2: 13, 14, 15, 16 and 17
- Group 3: 25, 26, 27, 28 and 29
- Group 4: 37, 38, 39, 40 and 41
Females:
- Group 1: 60, 63, 64, 66 and 67
- Group 2: 82, 74, 76, 83 and 79
- Group 3: 84, 85, 86, 87 and 88
- Group 4: 98, 99, 100, 101 and 102

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood:
- Animals fasted: Yes
- How many animals: same as 'haematology'

NEUROBEHAVIOURAL EXAMINATION: Yes
- Dose groups that were examined: all
- Battery of functions tested: sensory activity, grip strength, motor activity,

Thyroid hormone analysis: T4 levels.
Day 4 of age Offspring: 2 females per litter
Day 13 of age Offspring: 2 males and 2 females per litter

Sacrifice and pathology:

GROSS PATHOLOGY: Yes (see table 1)

HISTOPATHOLOGY: Yes (see table 2)

Other examinations:

not specified

Statistics:

yes

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):

not examined

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

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:

no effects observed

Neuropathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not examined

Other effects:

no effects observed

Key result

Dose descriptor:

NOEL

Effect level:

ca. 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

behaviour (functional findings)

body weight and weight gain

clinical biochemistry

clinical signs

food consumption and compound intake

gross pathology

haematology

histopathology: non-neoplastic

mortality

organ weights and organ / body weight ratios

Critical effects observed:

not specified

Conclusions:

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.

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:

Guideline study Klimisch 1

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

Justification for classification or non-classification

PV3:4 is not classified for repeat dose toxicity and a STOT RE is not assigned