2-[(4-methyl-2-nitrophenyl)azo]-3-oxo-N-phenylbutyramide

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 19 FEB 1979 to 03 APR 1979

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: basic data given, comparable to guideline

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

1979

Report date:

1979

Reference 2

Reference Type:

publication

Title:

Subacute Toxicity Studies of Selected Organic Colorants.

Author:

Leist KH

Year:

1982

Bibliographic source:

Ecotoxicology and Environmental Safety 6: 457-463

Report date:

1982

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Deviations:

not applicable

GLP compliance:

no

Remarks:

study was performed previous to GLP implementation

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2-[(4-methyl-2-nitrophenyl)azo]-3-oxo-N-phenylbutyramide

EC Number:

219-730-8

EC Name:

2-[(4-methyl-2-nitrophenyl)azo]-3-oxo-N-phenylbutyramide

Cas Number:

2512-29-0

Molecular formula:

C17H16N4O4

IUPAC Name:

2-[(4-methyl-2-nitrophenyl)diazenyl]-3-oxo-N-phenylbutanamide

Test material form:

solid: nanoform, no surface treatment

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Strain: WISKf (SPF 71)
- Source: in house breeding; Hoechst, breeding colony, Hattersheim
- Weight at study initiation: males: 73.8 +/- 2.29 g (69 g - 79 g); females: 73.6 +/- 2.41 g (69 g - 78 g)
- Fasting period before study: no
- Housing: 5 animals/sex/cage
- Diet: Standard diet Altromin 1324 (Altromin GmbH, Lage, Germany), ad libitum
- Water: tap water, ad libitum
- Acclimation period: no data


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-28
- Humidity (%): 25-50

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: suspension in starch mucilage

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
- the test item suspension was freshly prepared every day before dosing
- 10% test item suspension in 2% starch mucilage, which was permanently stirred during the dosing procedure

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

30 days

Frequency of treatment:

daily except on weekends, i.e. 22 doses over a period of 30 days

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 1000 mg/kg bw
Basis:
actual ingested

No. of animals per sex per dose:

20

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: no lethality and no signs of toxicity were observed after single oral application of 5000 mg/kg bw

Positive control:

none

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: Yes
- Time schedule for examinations: one week before the first treatment, on the days of treatment, and two times per week (monday and thursday) during the recovery period

FOOD CONSUMPTION AND COMPOUND INTAKE (no feeding study): Yes
- Food consumption for each animal determined: Yes
- mean daily diet consumption calculated as % body weight: Yes

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No data

WATER CONSUMPTION: Yes

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: weekly
- Dose groups that were examined: all

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at the end of the treatment period and at the end of the recovery period
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: 5 males and 5 females of each groupd
- Parameters examined: hematocrit, hemoglobin, erythrocytes count, total and differential leucocyte counts, median cell volume, median cell hemoglobin, platelet count, prothrombin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at the end of the treatment period and at the end of the recovery period
- Animals fasted: No data
- How many animals: 5 males and 5 females of each group
- Parameters examined: serum alkaline phosphatase (SAP), serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT)

URINALYSIS: Yes
- Time schedule for collection of urine: at the end of the treatment period
- Metabolism cages used for collection of urine: yes
- Animals fasted: yes
- Parameters examined: appearance, colour, protein, glucose, hemoglobin, bilirubin, pH-value, sediment


NEUROBEHAVIOURAL EXAMINATION: No
- weekly examinations for neurologic disturbances

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes (liver, spleen, adrenals, kidney)

Other examinations:

ORGAN WEIGHTS:
- liver, spleen, adrenals, kidney

Results and discussion

Results of examinations

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:

not specified

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

no effects observed

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

SGPT: statistically significant decrease in females, fully reversible during the recovery period, no histopathological findings, therefore not regarded to be adverse

Urinalysis findings:

no effects observed

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not examined

Details on results:

CLINICAL SIGNS AND MORTALITY
- no effects

BODY WEIGHT AND WEIGHT GAIN
- no differences between treated animals and controls

FOOD CONSUMPTION
-no effects

WATER CONSUMPTION
- no effects

OPHTHALMOSCOPIC EXAMINATION
- not examined

CLINICAL CHEMISTRY
- SGPT statistically significantly decreased in females at the end of the treatment period, effects were fully reversible at the end of the recovery period; these effects were not regarded to be adverse as no histopathological findings were observed
- no effects in males or on other endpoints in females

HAEMATOLOGY
- no pathological findings

URINALYSIS
- no effects
- no discolouration of the urine observable

NEUROBEHAVIOUR
- no neurologic disturbances

ORGAN WEIGHTS
- no effects

GROSS PATHOLOGY
- no effects

HISTOPATHOLOGY: NON-NEOPLASTIC
- no effects

Effect levels

Key result

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other:

Remarks on result:

not determinable due to absence of adverse toxic effects

Target system / organ toxicity

Key result

Critical effects observed:

not specified

Any other information on results incl. tables

The test item was excreted via faeces but not via urine (no discolouration of the urine).

Applicant's summary and conclusion

Conclusions:

The test item did not reveal any substance related toxic effects under the conditions tested. The NOAEL in this subacute oral study was 1000 mg/kg bw in male and female rats.

Executive summary:

Male and female Wistar rats (20 animals per sex and dose group) received 22 daily oral (gavage) doses of 0 or 1000 mg test item per kg bw over a period of 30 days and were partially (10 animals per sex and dose group) observed for another 14 days without treatment.

Cage side observations, food consumption, body weight development, haematology, clinical biochemistry, urine analysis, macroscopic investigations, organ weights and histopathology of selected organs did not reveal any substance related toxic effects. The NOAEL in this study was 1000 mg/kg bw.