Methyl anthranilate

Repeated dose toxicity: oral

Administrative data

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data is from publication

Data source

Reference

Reference 1

Reference Type:

publication

Title:

Repeated dose toxicity study in rats

Author:

E. C. HAGAN et. al.

Year:

1967

Bibliographic source:

Food and cosmetic toxicology, 1967

Reference 2

Reference Type:

review article or handbook

Title:

Repeated dose toxicity study of test chemical in rats

Author:

US National Library Of Medicine

Year:

2020

Bibliographic source:

HSDB (Hazardous Substances Data Bank); US national Library of Medicine,2020

Reference 3

Reference Type:

other: Secondary source

Title:

Repeated dose toxicity study of test chemical on rats

Author:

The Joint FAO/WHO Expert Committee on Food Additives (JECFA)

Year:

1980

Bibliographic source:

WHO Food Additives Series - 1980

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

13 weeks repeated oral toxicity study of test chemical in rats.

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Methyl o-aminobenzoate

Cas Number:

134-20-3

Molecular formula:

C8H9NO2

IUPAC Name:

Methyl o-aminobenzoate

Test material form:

liquid

Details on test material:

IUPAC name: 2-Aminobenzoic acid, methyl ester
Mol. formula: C8H9NO2
Molecular Weight: 151.1641 gm/mol
Smiles: c1(c(cccc1)N)C(OC)=O
InChI: 1S/C8H9NO2/c1-11-8(10)6-4-2-3-5-7(6)9/h2-5H,9H2,1H3

Test animals

Species:

rat

Strain:

Osborne-Mendel

Details on species / strain selection:

not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: not specified
- Females (if applicable) nulliparous and non-pregnant: [yes/no] not specified
- Age at study initiation: not specified
- Weight at study initiation: not specified
- Fasting period before study: not specified
- Housing: The animals were housed individually in wire cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: not specified

DETAILS OF FOOD AND WATER QUALITY:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): not specified
- Humidity (%): not specified
- Air changes (per hr): not specified
- Photoperiod (hrs dark / hrs light): not specified

IN-LIFE DATES: From: To:not specified

Administration / exposure

Route of administration:

oral: gavage

Details on route of administration:

stomach tube

Vehicle:

corn oil

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- Rate of preparation of diet (frequency): Fresh diets were made and distributed weekly
- Mixing appropriate amounts with (Type of food): The concentration of the solution was adjusted for each level so that all rats received a constant volume of 1 ml of solution/kg daily
- Storage temperature of food: Not specified

VEHICLE
- Justification for use and choice of vehicle (if other than water): The test chemical was soluble in corn oil
- Concentration in vehicle: Not specified
- Amount of vehicle (if gavage): Not specified
- Lot/batch no. (if required): Not specified
- Purity:Not specified

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

Daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day (nominal)

Remarks:

0 ppm

Doses / concentrations 2

Dose / conc.:

50 mg/kg bw/day (nominal)

Remarks:

1000 ppm

Doses / concentrations 3

Dose / conc.:

500 mg/kg bw/day (nominal)

Remarks:

10000 ppm

No. of animals per sex per dose:

Total: 40 animals
Test group: 10 males and 10 females
Control group: 10 males and 10 females

Control animals:

yes, concurrent vehicle

Details on study design:

Not specified

Positive control:

not specified

Examinations

Observations and examinations performed and frequency:

The rat's weight, food intake and general condition were recorded every week. Haematological examinations were made at 3, 6, 12 and 22 months in the chronic experiments. These examinations included white cell counts, red cell counts, haemoglobins and haematocrits.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes
At the termination of the experiments the rats were sacrificed and exsanguinated

Other examinations:

The tissues of all the rats were examined macroscopically at the time of sacrifice. The viscera were removed and the liver, kidneys, spleen, heart, and testes were weighed. These organs, the remaining abdominal and thoracic viscera, and one hind leg, for bone, bone marrow, and muscle, were preserved in 10 ~o buffered formalin-saline solution for histopathological examination. For routine histopathology, sections were embedded in paraffin wax and stained with haematoxylin and eosin. Tissues from rats dying during the experiment were examined for gross changes and were preserved if autolysis was not advanced. Organs were
not weighed but abnormalities and the suspected reason for death were noted.

Statistics:

not specified

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):

not specified

Ophthalmological findings:

not specified

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Effect levels

Key result

Dose descriptor:

NOAEL

Effect level:

500 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

clinical signs

food consumption and compound intake

haematology

mortality

Remarks on result:

not determinable due to absence of adverse toxic effects

Target system / organ toxicity

Critical effects observed:

no

Applicant's summary and conclusion

Conclusions:

No Observed Adverse Effect Level (NOAEL) is considered to be 500 mg / kg body weight when male and female rats were orally treated with test substance for chronic study.

Executive summary:

13 weeks repeated oral toxicity study was performed on rats to determine the toxic nature of test chemical. The study was performed on 10 male and 10 female Osborne-Mendel rats, a control group containing the same number of rats as the test groups was included with test compound. The animals were housed individually in wire cages and had access to food and water at all times. The test chemical was administered via stomach tube in corn oil solution at a dose range of 0, 1,000 and 10,000 ppm (approximately equivalent to 50 and 500 mg/kg bw). The concentration of the solution was adjusted for each level so that all rats received a constant volume of 1 ml of solution/kg daily. The control animals were given an equal volume of corn oil. The rat's weight, food intake and general condition were recorded every week. Hematological examinations were made at 3, 6, 12 and 22 months. These examinations included white cell counts, red cell counts, hemoglobin and hematocrits. At the termination of the experiments the rats were sacrificed and exsanguinated. The tissues of all the rats were examined macroscopically at the time of sacrifice. The viscera were removed and the liver, kidneys, spleen, heart, and testes were weighed. These organs, the remaining abdominal and thoracic viscera, and one hind leg, for bone, bone marrow, and muscle, were preserved in 10 % buffered formalin-saline solution for histopathological examination. For routine histopathology, sections were embedded in paraffin wax and stained with haematoxylin and eosin. Tissues from rats dying during the experiment were examined for gross changes and were preserved if autolysis was not advanced. Organs were not weighed but abnormalities and the suspected reason for death were noted. After 13 weeks, No adverse effects were observed on body weights, appearance, food intake, hematology, organ weights, macroscopic or microscopic (at highest dietary level) examination of the major organs. Pathological changes attributable to disease or age were observed. Thus based on observations, No Observed Adverse Effect Level (NOAEL) is considered to be 500 mg / kg body weight indicating that the test chemical is not toxic in nature.