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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
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

Referenceopen allclose all

Reference Type:
publication
Title:
Screening of fragrance materials for allergenicity in the guinea pig I. Comparison of four testing methods
Author:
G. KLECAK, H. GELEICK, and J. R. FREY
Year:
1977
Bibliographic source:
J. Soc. Cosmet. Chem., 2 8, 53-64 (February 1977)
Reference Type:
secondary source
Title:
Opinion on Methyl-N-methylanthranilate
Author:
European Commission's scientific committee on consumer safety
Year:
2011
Bibliographic source:
EC Directorate - General for Health & Consumers scientific committee on consumer safety Opinion on Methyl-N-methylanthranilate

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: Open Epicutaneous test
Principles of method if other than guideline:
An Open Epicutaneous Test (OET) was performed on guinea pigs to assess the skin sensitization potential of the test chemical
GLP compliance:
not specified
Type of study:
open epicutaneous test
Justification for non-LLNA method:
Currently no LLNA study is available for assessment. The Guinea Pig Maximization Test (GPMT) has been carried out as an animal test to predict human sensitization for over a decade and is recommended by international test guidelines such as OECD.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
- Name of test material (as cited in study report): Methyl N-methyl anthranilate
- Molecular formula : C9H11NO2
- Molecular weight : 165.191 g/mole
- Substance type: Organic
- Physical State: Liquid
Specific details on test material used for the study:
- Name of test material (as cited in study report): Methyl N-methyl anthranilate
- Molecular formula : C9H11NO2
- Molecular weight : 165.191 g/mole
- Substance type: Organic
- Physical State: Liquid

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Himalayan white-spotted
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Institute Of Biomedical Research Fullinsdrof Switzerland
- Age at study initiation:
- Weight at study initiation: 400 to 500 g.
- Housing:
- Diet (e.g. ad libitum): supplemented with green,Vegetables, carrots and vitamin C ad libitum
- Water (e.g. ad libitum): ad libitum

Study design: in vivo (non-LLNA)

Induction
Route:
epicutaneous, open
Vehicle:
other: ethanol, acetone, H20, petroleum, PEG and/or other suitable vehicles
Concentration / amount:
Concentration:100%, 30%, 10%, 3%, 1%, or 0.3%
Amount: 0.1ml
Day(s)/duration:
3 weeks
Adequacy of induction:
not specified
Challenge
No.:
#1
Route:
epicutaneous, open
Vehicle:
other: ethanol, acetone, H20, petroleum, PEG and/or other suitable vehicles
Concentration / amount:
Concentration:3%
Amount: 0.025ml
Day(s)/duration:
on days 21 and 35
Adequacy of challenge:
not specified
No. of animals per dose:
6-8 guinea pigs
Details on study design:
RANGE FINDING TESTS: The minimal irritating concentration of each material is used to confirm the biological activity determined before starting the induction. These tests are performed by applying with a pipette 0.025 ml of each concentration to skin areas measuring 2 cm2. The reactions are read after 24, 48 and/or 72h.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 21
- Exposure period:24 hours
- Test groups:6-8 guinea pigs
- Control group:3 guinea pigs
- Site: an area measuring 8 cm2 on the clipped flank skin of the guinea pigs
- Frequency of applications: The applications are repeated daily for 3 weeks or done 5 times weekly during 4 weeks
- Duration: 21 days (3 weeks)
- Concentrations: 100%, 30%, 10%, 3%, 1%, or 0.3% in vehicle.
Amount: 0.1ml

B. CHALLENGE EXPOSURE
- No. of exposures: 2
- Day(s) of challenge: on days 21 and 35
- Exposure period:24 hours
- Test groups: 6-8 guinea pigs
- Control group: 3 guinea pigs
- Site: contralateral flank measuring 2 cm2
- Concentrations: Concentration:6%
Amount: 0.025ml
- Evaluation (hr after challenge): 24,48 and/or 72h.
Challenge controls:
The 10 controls were either left untreated or treated with 0.1 ml aliquot of the vehicle for 21 days
Positive control substance(s):
not specified

Results and discussion

In vivo (non-LLNA)

Results
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
3%
No. with + reactions:
0
Total no. in group:
8
Clinical observations:
It was observed that none of the guinea pigs induced contact sensitization at challenge concentration of 3%.
Remarks on result:
no indication of skin sensitisation

Any other information on results incl. tables

Table: Allergenicity of Compounds tested in Humans by the Maximization Test and in Guinea Pigs by 4 Different Procedures

Guinea pigs

OET

DTb

MTb

FCATb

Minimum irritating concentration

(%)

Results

Results

Results

Results

After 1 day application

After 21 days application

100

3

-

-

-

-

a- the occlusive eliciting concentration application was only at the user concentration *2              

b- DT, MT, FCAT: Concentrations see in the method

 

Applicant's summary and conclusion

Interpretation of results:
other: not sensitizing
Conclusions:
It was observed that the test chemical failed to induce contact sensitization at challenge concentration of 3% in the test animals. Thus, the test chemical was considered to be not sensitizing on skin of guinea pigs at concentration of 3% in an Open Epicutaneous Test (OET).
Executive summary:

An Open Epicutaneous Test (OET) was performed on guinea pigs to assess the skin sensitization potential of the test chemical.

 

On day 1 during induction, 0.1 ml of the test chemical was applied at concentrations of 100%, 30%, 10%, 3%, 1%, or 0.3% in vehicle to an area measuring 8 cm2 on the clipped flank skin of the guinea pigs. The applications are repeated daily for 3 weeks or done 5 times weekly during 4 weeks, usually on the same skin sites. The application sites were left uncovered and the reactions, if continuous daily applications were performed, can be read 24 h after each application, or at the end of each week.

 

To determine whether or not contact sensitization was induced, all groups of guinea pigs previously treated for 21 days, as well as 10 untreated, or only pretreated with the vehicle, controls are tested on days 21 and 35 on the contralateral flank with the test material. This test was performed by applying with a pipette 0.025 ml of each concentration to skin areas measuring 2 cm2. The reactions were read after 24, 48 and/or 72h.

 

It was observed that the test chemical failed to induce contact sensitization at challenge concentration of 3% in the test animals. Thus, the test chemical was considered to be not sensitizing on skin of guinea pigs at concentration of 3% in an Open Epicutaneous Test (OET).