Methyl N-methylanthranilate

Toxicity to reproduction

Administrative data

Endpoint:

toxicity to reproduction

Remarks:

other: Chronic repeated dose toxicity

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 peer- reviewed journal

Data source

Reference

Reference Type:

publication

Title:

Acute and Short-term Toxicity of Methyl-N-methyl Anthranilate in Rats

Author:

I. F. Gaunt and M. Sharra TT P. Grasso and M. Wright

Year:

1970

Bibliographic source:

Food Cosmet. Toxicol. Vol. 8, pp. 359-368. Pergamon Press 1970.

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: as below

Principles of method if other than guideline:

Chronic repeated dose toxicity study of tets material in rats.

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Methyl N-methylanthranilate

EC Number:

201-642-6

EC Name:

Methyl N-methylanthranilate

Cas Number:

85-91-6

Molecular formula:

C9H11NO2

IUPAC Name:

methyl 2-(methylamino)benzoate

Constituent 2

Reference substance name:

Methyl -N-methylanthranilate

IUPAC Name:

Methyl -N-methylanthranilate

Details on test material:

- Name of test material (as cited in study report): Methylanthranilate (MMA)
- Molecular formula (if other than submission substance): C9H11NO2
- Molecular weight (if other than submission substance): 165.191 /mole
- Substance type: Organic
- Physical state: solid
- Impurities (identity and concentrations): 2 %

Test animals

Species:

rat

Strain:

other: CFE

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: SPF colony
- Age at study initiation: (P) x wks; (F1) x wks: No data available
- Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x g: 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

Administration / exposure

Route of administration:

oral: feed

Vehicle:

other: Spillers' Laboratory Small Animal Diet

Details on exposure:

No data available

Details on mating procedure:

No data available

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

By using Pye 104 dual flame ionization chromatograph.

Duration of treatment / exposure:

90 days

Frequency of treatment:

Daily

Details on study schedule:

No data available

Doses / concentrations

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:
actual ingested

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

Positive control:

No data available

Examinations

Parental animals: Observations and examinations:

Clinical sign, Body Weight, Food consumption, Water Consumption, Haematology, Clinical Chemistry and Urinalysis were observed .

Oestrous cyclicity (parental animals):

No data available

Sperm parameters (parental animals):

No data available

Litter observations:

No data available

Postmortem examinations (parental animals):

Reporductive organ weight, gross pathology and histopathology were exmined.

Postmortem examinations (offspring):

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.

Reproductive indices:

No data available

Offspring viability indices:

No data available

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Dermal irritation (if dermal study):

not specified

Mortality:

not specified

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:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not specified

Other effects:

no effects observed

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

not specified

Details on results (P0)

Mortality: No data available

Clinical signs:No signs of ill health were observed in treated rat during the study as compared to control.

Body weight: No effect was observed on body weight and weight gain of treated rat as compared to control.

Food consumption: No effect was observed on food consumption of treated rat as compared to control.

Test substance 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.

Organ weights: When male and female rats treated with 244 and 280 mg/kg bw, Absolute and relative kidney weight was significantly increased in male and relative weight in female as compared to control.
When male and female rats treated with 82 and 95 mg/kg bw, Absolute and relative kidney weight was significantly increased in male and relative kidney, adrenals and ovaries weight in female as compared to control.
No effect on reproductive oragn weight of female gonads were observed at 288 mg/kg bw.

Gross pathology: No gross abnormalities were observed in Gonads of treated male and female rat as compared to control.

Histopathology: No histopathological changes were observed in Gonads of treated male and female rat as compared to control.

other findings:
Water consumption: No effect was observed on water consumption of treated rat as compared to control.

Haematology:
At 6 week,
When treated with 244 and 280 mg/kg bw, 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 82 and 96 mg/kg bw, 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 chemistry:No significant differences were observed in treated rat as compared to control.

Urinanalysis: No significant differences were observed in urine analyses of treated rat as compared to control.

Effect levels (P0)

1

Dose descriptor:

NOAEL

Effect level:

244 mg/kg bw/day

Based on:

test mat.

Sex:

male

Basis for effect level:

other: No effect on reporductive organweight, gorss pathology and histopathology

Remarks on result:

other: No effect on reporductive organ weight

2

Dose descriptor:

NOAEL

Effect level:

288 mg/kg bw/day

Based on:

test mat.

Sex:

female

Basis for effect level:

other: No effect on reporductive organ weight, gorss pathology and histopathology

Remarks on result:

other: No effect on reporductive organ weight

Target system / organ toxicity (P0)

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Results: F1 generation

General toxicity (F1)

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality / viability:

not specified

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Histopathological findings:

not specified

Other effects:

not specified

Developmental neurotoxicity (F1)

Behaviour (functional findings):

not specified

Developmental immunotoxicity (F1)

Developmental immunotoxicity:

not specified

Effect levels (F1)

Dose descriptor:

other: not specified

Generation:

other: not specified

Based on:

not specified

Sex:

not specified

Basis for effect level:

other: not specified

Remarks on result:

not measured/tested

Target system / organ toxicity (F1)

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Overall reproductive toxicity

Reproductive effects observed:

not specified

Treatment related:

not specified

Relation to other toxic effects:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Any other information on results incl. tables

Absolute and relative organ weights of rats fed at 0-3600 ppm of the diet for 13 wk

Sex and dietary level (ppm)	Organ weights												Terminal body weight (g)
	Brain	Heart	Liver	Spleen	Stomach	Intestine	Caecum	Kidneys	Adrenalst	Gonads	Pituitaryt	Thyroid
Male	Absolute organ weight (g)
0	1.88	1.29	10.31	0.75	1.52	5.67	1.07	2.79	55	3.62	12	22	415
300	1.87	1.31	10.24	0.72	1.52	5.96	1.09	2.92	52	3.53	12	22	420
1200	1.85	1.30	10.49	0.77	1.50	5.84	1.01	2.97**	54	3.57	12	23	42
3600	1"86	1-28	10.57	0'75	1.56	5.76	1.16	3.04**	53	3.47	12	22	414
Female
0	1.76	0.96	0.62	1.38	5.43	0.89	0.89	1.83	61	0.13	15	19	286
300	1.74	0.93	6.35	0-58	1.36	5.60	0.92	1.85	62	0.14	15	19	283
1200	1.76	0.93	6.25	0.60	1.32	5.34	0.89	1.88	66	0.15	15	20	27
3600	1.87	0.93	6.58	0.59	1.33	5.66	0.94	1.91	64	0.13	16	19	282
Male	Relative organ weight (g/100 g body weight
0	0.45	0.31	2.50	0.18	0.37	1.37	0.25	0.67	13.2	0.87	2.82	5.26
300	0.44	0.31	2.44	0.17	0.36	1.42	0.25	0.69	12.4	0.84	2.85	5.22
1200	0.44	0.31	2.47	0.18	0.35	1.38	0.23	0.70*	12.8	0.84	2.82	5.52
3600	0.45	0.31	2.57	0.18	0.38	1.39	0.28	0.73***	12.8	0.84	2.77	4.99
Female
0	0.62	0.33	2.40	0.21	0.48	1.89	0.32	0.64	21.6	0.045	5.17	6.62
300	0.62	0.33	2.37	0.21	0.48	1.97	0.32	0.66	21.8	0.048	5.23	6.56
1200	0.64	0.34	2.42	0.22	0.48	1.94	0.32	0.69**	24.1'*	0.052*	5.53	7.10
3600	0.63	0.33	2.47	0.21	0.47	2.00	0.33	0.68*	22.7	0.046	5.68	6.56

ϮValues of absolute and relative weights of this organ are expressed in mg and mg[100 g body weight respectively.

Values are the means of groups of 15 animals and those marked with asterisks differ significantly (Student'sttest) from those of controls: *P < 0"05; **P < 0"01; ***P < 0"001.

Applicant's summary and conclusion

Conclusions:

NOAEL was considered to be 244 mg/kg body weight/day for male and 288 mg/kg bw for female when CFE male and female rat treated with test material orally.

Executive summary:

In a Chronic repeated dose toxicity study, CFE male and female rat were treated with test material in the concentration of 0, 300, 1200 and 3600 ppm orally in diet equivalant to average intakes of 21, 82 and 244 mg/kg/day for males and 24, 95 and 280 mg/kg/ day for females.No signs of ill health, change in body weight,food and water consumption, Clinical chemistry and Urinanalysis were observed in treated male and female rat during the study as compared to control. At 6 week, significant decrease was observed in hemoglobin, RBC and WBC (particularly the lymphocytes) of male and female rat when treated with 82 and 244 mg/kg bw and 82 and 96 mg/kg bw treated rats. 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 82 and 244 mg/kg bw and 82 and 96 mg/kg bw treated rats as compared to control. As no effect on reproductive oragn weight of female gonads were observed in 288 mg/kg bw, the observed change in relative organ weight of female gonads were considered to be non significant. In addition,No gross abnormalities and histopathological changes were observed in Gonads of treated male and female rat as compared to control.Therefore, NOAEL was considered to be 244 mg/kg body weight/day for male and 288 mg/kg bw for female when CFE male and female rat treated with test material orally in feed for 90 days.