Registration Dossier

Administrative data

Description of key information

Repeated dose toxicity: via oral route;

The No Observed Adverse Effect Level (NOAEL) was considered to be 300 ppm (20 mg/kg body weight/day) when CFE male and female rat treated with test substance for 90 days by oral feed.

Repeated inhalation study:

According to Annex IX of the REACH regulation, testing by the inhalation route is appropriate only if exposure of humans via inhalation is likely. Taking into account the low vapour pressure of the substance Methyl 2-(methylamino)benzoate (85-91-6) which is reported as 0.02055169 mmHg at 25 C. .Thus, exposure to inhalable dust, mist and vapour of the chemical Methyl 2-(methylamino)benzoate is highly unlikely. Therefore this study is considered for waiver.

Repeated dermal study;

The acute toxicity value for Methyl 2-(methylamino)benzoate (85-91-6) (as provided in section 7.2.3) is >2000 mg/kg body weight. Also, given the use of the chemical; repeated exposure by the dermal route is unlikely since the use of gloves is common practice in industries. Thus, it is expected that Methyl 2-(methylamino)benzoate shall not exhibit 28 day repeated dose toxicity by the dermal route. In addition, there is no data available that suggests that Methyl 2-(methylamino)benzoate shall exhibit repeated dose toxicity by the dermal route. Hence this end point was considered for waiver.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Data is from peer reviewed journal
Justification for type of information:
Data is from peer reviewed journal
Qualifier:
according to
Guideline:
other: Chronic repeated dose toxicity study of Methyl N-methylanthranilate (MMA) in rat orally was performed for 90 days
Principles of method if other than guideline:
Chronic repeated dose toxicity study of test chemical in rat orally was performed for 90 days
GLP compliance:
no
Limit test:
no
Species:
rat
Strain:
other: CFE
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: SPF colony
- Age at study initiation: No data available
- Weight at study initiation: No data available
- Fasting period before study: No data available
- Housing: Animals were housed 5 rat per cage
- Diet (e.g. ad libitum): Spillers' Laboratory Small Animal Diet, ad libitum
- Water (e.g. ad libitum): Water, ad libitum
- Acclimation period: No data available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): No data available
- Humidity (%):No data available
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): No data available

IN-LIFE DATES: From: To: No data available
Route of administration:
oral: feed
Vehicle:
other: Spillers' Laboratory Small Animal Diet
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: No data available

DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): Spillers' Laboratory Small Animal Diet
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): Spillers' Laboratory Small Animal Diet
- Concentration in vehicle: 0, 300, 1200 and 3600 ppm
- Amount of vehicle (if gavage): No data available
- Lot/batch no. (if required): No data available
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Yes, By using Pye 104 dual flame ionization chromatograph.
Duration of treatment / exposure:
90 days
Frequency of treatment:
Daily
Remarks:
Doses / Concentrations:
0, 300, 1200 and 3600 ppm (21, 82 and 244 mg/kg/day for the three dose levels in males and 24, 95 and 280 mg/kg/day in females.
Basis:
nominal in diet
No. of animals per sex per dose:
Total: 120
0 ppm : 15 male, 15 female
300 ppm : 15 male, 15 female
1200 ppm : 15 male, 15 female
3600 ppm : 15 male, 15 female
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Because of the small amount of flavoring lost from the test diets and its apparent palatability, the material was administered in the diet at 0, 300, 1200 and 3600 ppm
- Rationale for animal assignment (if not random): No data available
- Rationale for selecting satellite groups: No data available
- Post-exposure recovery period in satellite groups: No data available
- Section schedule rationale (if not random): No data available
Positive control:
No data
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: No data available
- Time schedule: No data available
- Cage side observations checked in table [No.?] were included. No data available

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Weekly

BODY WEIGHT: Yes
- Time schedule for examinations:
:Weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data available - Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data available

FOOD EFFICIENCY: No data available
- 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 available


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: water intake was measured over a 3-day period during wk 4, 8 and 12.

OPHTHALMOSCOPIC EXAMINATION: No data available
- Time schedule for examinations: No data available
- Dose groups that were examined: No data available

HAEMATOLOGY: Yes
- Time schedule for collection of blood: At week 6 and at the time of autopsy.
- Animals fasted: No data available
- How many animals: No data available
- Parameters checked in table were examined.: Yes
The haemoglobin content, packed cell volume and erythrocyte, reticulocyte and total and differential leucocyte counts were determined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: At autopsy
- Animals fasted: No data available
- How many animals: No data available
- Parameters checked in table [No.?] were examined: Urea nitrogen, glucose, activities of glutamic-oxalacetic and glutamic-pyruvic transaminases were examined.

URINALYSIS: Yes
- Time schedule for collection of urine: During wk 6 and 13.
- Metabolism cages used for collection of urine: No data available
- Animals fasted: No data available
- Parameters checked in table [No.?] were examined: Colour, glucose, bile salts, blood, ketones and microscopic constituents were examined.

Urine dilution and concentration tests were conducted on the same rats, the volume and specific gravity of urine produced in 2 hr after a water load of 25 ml/kg and after 6 hr and 16 hr of water deprivation being measured.

NEUROBEHAVIOURAL EXAMINATION: No data available

OTHER:
Organ weight: Yes
Brain, pituitary, thyroid, heart, liver, spleen, stomach, small intestine, caecum, adrenals, kidneys and gonads were weighted.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
Macroscopic abnormalities were noted.

Organ and tissue were preserved in 10 % buffered formalin.

Paraffin wax sections of these tissues were stained with haematoxylin and eosin for microscopic examination.

HISTOPATHOLOGY: Yes

Organ examined.
Brain, pituitary, thyroid, heart, liver, spleen, stomach, small intestine, caecum, adrenals, kidneys and gonads
lymph nodes, thymus, urinary bladder, colon, rectum, pancreas, uterus and muscle were examined
Other examinations:
No data available
Statistics:
Statistical analysis is performed by using Student's t test for the significance of Haematological examination and Absolute and relative organ weights
Clinical signs:
no effects observed
Description (incidence and severity):
No signs of ill health were observed in treated rat during the study as compared to control.
Mortality:
no mortality observed
Description (incidence):
No signs of ill health were observed in treated rat during the study as compared to control.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No effect was observed on body weight and weight gain of treated rat as compared to control.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No effect was observed on food consumption of treated rat as compared to control. Compound intake: Average intakes of were 21, 82 and 244 mg/kg/day for the three dose levels in males and 24, 95 and 280 mg/kg/day in females.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
No effect was observed on water consumption of treated rat as compared to control.
Ophthalmological findings:
not specified
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
Hematology: At 6 week, When treated with 3600 ppm, in male rat significant decrease were observed in hemoglobin, RBC and WBC (particularly the lymphocytes) and in female significant decrease hemoglobin were observed. When treated with 1200 ppm, in male rat significant decrease were observed in WBC (particularly the lymphocytes) and in female hemoglobin level as compared to control. At autopsy, no significant differences between treated and control groups were observed
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
No significant differences were observed in treated rat as compared to control.
Urinalysis findings:
no effects observed
Description (incidence and severity):
No significant differences were observed in urine analyses of treated rat as compared to control.
Behaviour (functional findings):
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
When treated with 3600 ppm, Absolute and relative kidney weight was significantly increased in male and relative weight in female as compared to control. When treated with 1200 ppm, Absolute and relative kidney weight was significantly increased in male
Gross pathological findings:
no effects observed
Description (incidence and severity):
No gross abnormalities were observed in treated rat as compared to control.
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
No histopathological changes were observed in treated rat as compared to control.
Key result
Dose descriptor:
NOAEL
Effect level:
ca. 21 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: No adverse effect on clinical sign, body weight, food consumption, water consumption, haematology, clinical chemistry, urinanalysis, organ weights, gross pathology and histopathology.
Key result
Dose descriptor:
NOAEL
Effect level:
24 other: mg/kg/day
Based on:
test mat.
Sex:
female
Basis for effect level:
other: No significant effect were observed at this dose
Remarks on result:
other: No toxic effects were observed
Critical effects observed:
not specified

The slight reduction in haemoglobin content and red cells found in the peripheral blood of male and female rats and the reduction in the numbers of white cells found in males during week 6 were not found in blood taken from the aorta at autopsy.

 

Although statistically significant, the elevation in kidney weight in rats on the two highest dosage levels was less than 10 % of the control kidney weight. It was not accompanied by any detectable changes in the renal function tests or by macroscopic or microscopic damage.

Conclusions:
The No Observed Adverse Effect Level (NOAEL) was considered to be 300 ppm (21 and 24 mg/kg body weight/dayfor male and female respectivley) when CFE male and female rat treated with test substance for 90 days by oral feed.
Executive summary:

In a Chronic repeated dose toxicity study, CFE male and female rat were treated with test chemical in the concentration of 0, 300, 1200 and 3600 ppm orally in diet. At 6 week, significant decrease was observed in hemoglobin, RBC and WBC (particularly the lymphocytes) of male and female rat when treated with 1200 and 3600 ppm treated rat. Similarly, Absolute and relative kidney weight was significantly increased in male rat and absolute and relative kidney weight , relative adrenals and ovaries weight in female when treated with 1200 and 3600 ppm treated rat as compared to control. Therefore, the No Observed Adverse Effect Level (NOAEL) was considered to be 300 ppm (21mg/kg/day for male and 24mg/kg/day for female body weight/day) when CFE male and female rat were treated with test chemical orally in diet for 90 days.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
24 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
Data is from qualified publication.

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

The data available for the test chemical was reviewed to determine the toxic nature methyl 2-(methylamino)benzoate (85-91-6) repeated exposure by oral route. The study is as mentioned below:

Repeated dose toxicity: via oral route;

In a Chronic repeated dose toxicity study, CFE male and female rat were treated with test chemical in the concentration of 0, 300, 1200 and 3600 ppm orally in diet. At 6 week, significant decrease was observed in hemoglobin, RBC and WBC (particularly the lymphocytes) of male and female rat when treated with 1200 and 3600 ppm treated rat. Similarly, Absolute and relative kidney weight was significantly increased in male rat and absolute and relative kidney weight , relative adrenals and ovaries weight in female when treated with 1200 and 3600 ppm treated rat as compared to control. Therefore, the No Observed Adverse Effect Level (NOAEL) was considered to be 300 ppm (21mg/kg/day for male and 24mg/kg/day for female body weight/day) when CFE male and female rat were treated with test chemical orally in diet for 90 days.

Supported by other repeated dose toxicity study. The activity of test chemical was studied by a short-term (90 days) feeding studies in rats. Single dosage levels for each substance were derived from the total estimated daily intake, calculated on a mg/kg body weight basis assuming 50 kg as the average body weight, and multiplying by 100. The test material was given orally through diet to 15/sex FDRL strain rats at a dose concentration of 0 or 20.3 mg/Kg bw/day. The actual ingested dose was found to be 19.9 mg/Kg bw/day for males and 22.2 mg/Kg bw/day for females. Various observations were conducted and the NOAEL value was found to be19.9 mg/Kg bw/day in males and 22.2 mg/Kg bw/day in females. No significant gross pathological change was observed at autopsy in any of the rats, with the exception of occasional pulmonary alterations associated with a respiratory infection. None of the deviations shown for the test groups can be regarded as pathological even though they fall somewhat outside the statistical limits (P=0.05) for the controls and were not dose related. 90-Day feeding studies in groups of rats receiving dietary doses of test chemical at levels corresponding to at least 100 times the maximum estimated human dietary levels, revealed no evidence of adverse toxic effects.

Based on the data available from the test chemical methyl 2-(methylamino)benzoate (85-91-6) does not exhibit repeated dose oral toxicity in the dose range of 22.2-24 mg/kg bw/day for females and 19.9 -21 mg/kg/day for males respectively. Hence the test chemical is not likely to classify as a repeated dose oral toxicity as per the criteria mentioned in CLP regulation.

Repeated inhalation study:

According to Annex IX of the REACH regulation, testing by the inhalation route is appropriate only if exposure of humans via inhalation is likely. Taking into account the low vapour pressure of the substance Methyl 2-(methylamino)benzoate (85-91-6) which is reported as 0.02055169 mmHg at 25 C. .Thus, exposure to inhalable dust, mist and vapour of the chemical Methyl 2-(methylamino)benzoate is highly unlikely. Therefore this study is considered for waiver.

Repeated dermal study;

The acute toxicity value for Methyl 2-(methylamino)benzoate (85-91-6) (as provided in section 7.2.3) is >2000 mg/kg body weight. Also, given the use of the chemical; repeated exposure by the dermal route is unlikely since the use of gloves is common practice in industries. Thus, it is expected that Methyl 2-(methylamino)benzoate shall not exhibit 28 day repeated dose toxicity by the dermal route. In addition, there is no data available that suggests that Methyl 2-(methylamino)benzoate shall exhibit repeated dose toxicity by the dermal route. Hence this end point was considered for waiver.

Based on the data available for the test chemical Methyl 2-(methylamino)benzoate (85-91-6) does not exhibit toxic nature upon repeated exposure by oral, inhalation and dermal route of exposure and hence is not likely to classify as per the criteria mentioned in CLP regulation.

Justification for classification or non-classification

Based on the data available for the test chemical Methyl 2-(methylamino)benzoate (85-91-6) does not exhibit toxic nature upon repeated exposure by oral, inhalation and dermal route of exposure and hence is not likely to classify as per the criteria mentioned in CLP regulation.