Methyl anthranilate

Endpoint summary

Description of key information

Skin irritation

Skin irritation study was performed on Human subjects to determine the irritating effects of test chemical. No irritation was observed after a 48-hr closed-patch test on human subjects and this result was confirmed when the test was repeated. Thus the test chemical was not irritating to human skin and cannot be classified as a Skin Irritant.

Eye irritation

Eye irritation study was performed according EPA OPP Series 81-4 to assess the irritation potential of the test chemical.The eye irritation scores after 72 hours were as follows- Cornea score -5/6, iris score – 4/6, conjunctivae redness score – 5/6, conjunctivae chemosis score -4/6, conjunctivae discharge – 2/6. Corneal Effects got cleared in 8 -21 days. Based on the test conditions, the test chemical was considered irritating to eyes. The eye irritation study was placed in TOX Category II according to EPA OPP regulations.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

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

Qualifier:

according to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

To determine the kin irritation potential of test chemical on Humans.

GLP compliance:

not specified

Species:

other: Human

Strain:

not specified

Details on test animals or test system and environmental conditions:

Not specified

Type of coverage:

occlusive

Preparation of test site:

not specified

Vehicle:

other: petrolatum

Controls:

not specified

Amount / concentration applied:

10%

Duration of treatment / exposure:

48 hours

Observation period:

Not specified

Number of animals:

Not specified

Details on study design:

Not specified

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

48 h

Score:

0

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

No irritaion observed.

Other effects:

Not specified

Interpretation of results:

other: Not irritating

Conclusions:

Skin irritation study was performed on Human subjects to determine the irritating effects of test chemical. No irritation was observed after a 48-hr closed-patch test on human subjects and this result was confirmed when the test was repeated. Thus the test chemical was not irritating to human skin and cannot be classified as a Skin Irritant.

Executive summary:

Skin irritation study was performed on Human subjects to determine the irritating effects of test chemical. The test chemical when tested at 10% in petrolatum, it produced no irritation after a 48-hr closed-patch test on human subjects and this result was confirmed when the test was repeated. Thus the test chemical was not irritating to human skin and cannot be classified as a Skin Irritant.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

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 safety assessment reports

Qualifier:

according to guideline

Guideline:

EPA OPP 81-4 (Acute Eye Irritation)

Deviations:

yes

Remarks:

The test procedure deviated from Series 81-4.

Principles of method if other than guideline:

To assess the ocular irritation potential of the test chemical according to EPA OPP 81-4 with slight deviation.

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

- Source: Davidson’s Mill Farm, S.Brunswick, NJ,USA
Standard laboratory animal husbandry were observed

Vehicle:

unchanged (no vehicle)

Controls:

not specified

Amount / concentration applied:

0.1 ml of Undiluted sample as a single instillation

Duration of treatment / exposure:

not specified

Observation period (in vivo):

Observation of the effects were made at 1,24,48 and 72 hours and days 4, 7, 10, 24

Number of animals or in vitro replicates:

no data

Details on study design:

Eyes were examined with 2% sodium fluorescein 24 hours prior to application of test chemical.
SCORING SYSTEM: Irritancy scores were made after Kay, et al (1962)

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

72 h

Score:

5

Max. score:

6

Reversibility:

fully reversible within: 8-21 days

Remarks on result:

positive indication of irritation

Irritation parameter:

iris score

Basis:

mean

Time point:

72 h

Score:

5

Max. score:

6

Reversibility:

fully reversible within: 14 days

Remarks on result:

positive indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Time point:

72 h

Score:

5

Max. score:

6

Reversibility:

fully reversible within: 14 days

Remarks on result:

positive indication of irritation

Irritation parameter:

chemosis score

Basis:

mean

Time point:

72 h

Score:

4

Max. score:

6

Reversibility:

fully reversible within: 14 days

Remarks on result:

positive indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

mean

Time point:

72 h

Score:

2

Max. score:

6

Reversibility:

fully reversible within: 14 days

Remarks on result:

positive indication of irritation

Irritant / corrosive response data:

Maximum score 6/6 was observed in conjunctivae irritation parameters after 1 hour .Corneal Effects got cleared in 8 -21 days

Table 1: Eye Irritation score card

	No of affected/ No of exposed in days
Ocular effects	1 hour	1	2	3	4	7	10	14
Cornea	1/6	6/6	6/6	5/6	5/6	3/6	2/6	0/6
Iris	2/6	4/6	4/6	4/6	3/6	1/6	0/6	0/6
Conjunctivae
Redness	6/6	6/6	6/6	5/6	5/6	1/6	0/6	0/6
Chemosis	6/6	6/6	5/6	4/6	0/6	0/6	0/6	0/6
Discharge	6/6	6/6	4/6	2/6	0/6	0/6	0/6	0/6

Interpretation of results:

other: Category 2 as per the CLP criteria of classification and labelling.

Conclusions:

Eye irritation study was performed according EPA OPP Series 81-4 to assess the irritation potential of the test chemical.The eye irritation scores after 72 hours were as follows- Cornea score -5/6, iris score – 4/6, conjunctivae redness score – 5/6, conjunctivae chemosis score -4/6, conjunctivae discharge – 2/6. Corneal Effects got cleared in 8 -21 days. Based on the test conditions, the test chemical was considered irritating to eyes. The eye irritation study was placed in TOX Category II according to EPA OPP regulations.

Executive summary:

Eye irritation study was performed according EPA OPP Series 81-4 to assess the irritation potential of the test chemical. The test procedure deviated from EPA OPP Series 81-4. Eyes were examined with 2% sodium fluorescein 24 hours prior to application of test chemical.0.1 ml of Undiluted sample as a single instillation.Observations of the effects were made at 1, 24, 48 and 72 hours and days 4, 7, 10, 24.Irritancy scores were made after Kay, et al (1962). The eye irritation scores after 72 hours were as follows-Cornea score -5/6, iris score – 4/6, conjunctivae redness score – 5/6, conjunctivae chemosis score -4/6, conjunctivae discharge – 2/6. Corneal Effects got cleared in 8 -21 days. Based on the test conditions, the test chemical was considered irritating to eyes. The eye irritation study was placed in TOX Category II according to EPA OPP regulations.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Additional information

Skin irritation:

Skin irritation study was performed on test chemical to determine the irritating potency of test chemical. The studies are summarized below:

Skin irritation study was performed on Human subjects to determine the irritating effects of test chemical. The test chemical when tested at 10% in petrolatum, it produced no irritation after a 48-hr closed-patch test on human subjects and this result was confirmed when the test was repeated. Thus the test chemical was not irritating to human skin and cannot be classified as a Skin Irritant.

Skin irritation study was performed on rabbits to deteremine the irritating effects of test chemical. The test chemical was applied full strength to intact or abraded rabbit skin for 24 hr under occlusion. Moderate irritating effects were observed indicating that the test chemical can be cotegorized as Category 2 (irritant) based on GHS criteria.

A study was designed and conducted to determine the dermal reaction profile of the test chemical in Sprague Dawley rats. The study was performed as per OECD Guidelines 402 and complying to the GLP procedures. Ten rats (5 male and 5 female) were used for conducting dermal irritation /corrosion study.

The animals were kept in their cages for at least 5 days prior to administration for acclimatization to the laboratory condition and after acclimatization period, animals were randomly selected. Approximately 24 hours before application, the hair of each rat was closely clipped from the trunk (dorsal surface and sides from scapular to pelvic area) with an electric clipper, so as to expose at least 10% of the body surface area. The test item was applied directly onto the exposed skin of the animal, taking care to spread the test item evenly over the entire area of approximately 10% of the total body surface area or as much of the area as can reasonably be covered. The test item was held in contact with the skin using a porous gauze dressing and non irritating tape around the animal to cover the exposure site for first 24 hours exposure period. Elizabethan collar was placed on each animal for first 24 hours after application of the test item. These collars prevent ingestion of test item. Following 24 hours of exposure, the wrapping was removed and the test site wiped free of excess test item. Distilled water was used to remove residual test item. The test chemical was applied to shorn skin of 5 male and 5 female animals at 2000 mg/kg body weight. Administration of the test item at 2000 mg/kg did not result in any skin reaction at the site of application during the study period of 14 days. Also, the erythema and edema score of rats was calculated as 0. Administration of the test item did not result in any signs of toxicity and mortality during the study period of 14 days. Animals exhibited normal body weight gain through the study period of 14 days. Gross pathological examination did not reveal any abnormalities attributable to the treatment. Hence, it was concluded that the test chemical was Non-Irritating to the skin of Sprague Dawley rats under the experimental conditions tested and classified as “Category-Not Classified” as per CLP Classification.

A pre-test for a human maximization study was performed to assess the irritation potential of the test chemical. 8% test chemical in petrolatum was applied under occlusion in a closed patch to the backs of 27 male volunteers for 48 hours and the effects were observed. The test chemical was not irritating to the skin of 27 male volunteers after 48 hours exposure. Hence, test chemical can be considered as not irritating to skin.

Based on the available data for the test chemical, it can be concluded that the target chemical is Not irritating to skin of test animals. Therefore the target chemical was estimated to be Not irritating to skin and it can be further classified under the category “Not irritating” as per CLP regulation.

Eye Irritation:

Data available for the test chemical was reviewed to determine the Eye irritation potential of test chemical, The studies are as mentioned below:

Eye irritation study was performed according EPA OPP Series 81-4 to assess the irritation potential of the test chemical. The test procedure deviated from EPA OPP Series 81-4. Eyes were examined with 2% sodium fluorescein 24 hours prior to application of test chemical.0.1 ml of Undiluted sample as a single instillation.Observations of the effects were made at 1, 24, 48 and 72 hours and days 4, 7, 10, 24.Irritancy scores were made after Kay, et al (1962). The eye irritation scores after 72 hours were as follows-Cornea score -5/6, iris score – 4/6, conjunctivae redness score – 5/6, conjunctivae chemosis score -4/6, conjunctivae discharge – 2/6. Corneal Effects got cleared in 8 -21 days. Based on the test conditions, the test chemical was considered irritating to eyes. The eye irritation study was placed in TOX Category II according to EPA OPP regulations.

The MatTek EpiOcular™ model was used to assess the potential ocular irritation of the test article by determining the viability of the tissues following exposure to the test article via MTT. The objective of this study was to assess the ocular irritation potential of test article. Tissues were exposed to test article and controls for ~6 hours, followed by a ~25 minute post-soak and approximately 18 hour recovery after the post-soak. The viability of each tissue was determined by MTT assay. The mean OD570 of the negative control tissues was 1.665 and 1.589, which met the acceptance criteria of greater than 0.8 and less than 2.5. The mean relative viability of the positive control tissues was 45.2 and 36.5, which met the acceptance criterion of less than 50%. The differences in viability between identically treated tissues were 0.03 to 5.60, which met the acceptance criterion of less than 50%. All controls passed the acceptance criteria for a valid study. The mean % tissue viability of test chemical was determined to be 57.5%. Hence, under the experimental test conditions it was concluded that test chemical was considered to be irritating to the MatTek EpiOcular Tissue Model OCL-200 and being classified as ‘’Irritating to eyes in Category 2”

An eye irritation test was conducted to assess the irritation potency of the test chemical. Normal albino rabbit eyes were selected on the basis of absence of grossly visible staining by a 5-percent aqueous solution of fluorescein sodium, flushed with distilled water 20 seconds after application. A 2 hour interval was given so that the eye could return back to normal condition. Then, 0.005 ml of the undiluted test material was applied to the center of the cornea while the lids were retracted. About one minute later, the lids were released. This procedure is necessary to prevent the removal of a portion of the dose by the very efficient wiping system of the lids before intimate contact has been made with the eye. 18 to 24 hours later, the eyes were examined in strong diffuse daylight, then stained with fluorescein, and the injury was scored. The test chemical was grouped under Grade 3 injury (0.1 ml undiluted gives injury of up to 5 points (0.5 ml gives over 5)) and Grade 9 injury (Excess of 1% solution gives injury up to 5 points (5% gives over 5)) when rabbit eyes were observed for injuries. The test chemical was known to have caused loss of vision or very slowly healing corneal burns. Based on these grades, it can be inferred that the test chemical was irritating to rabbit eyes.

Based on the observations made, the test chemical was “An Eye Irritant (Irritating to Eyes)"  to test animals. it was classified under the category "Category 2" as per  CLP regulation.

Justification for classification or non-classification

The available studies indicate that the test chemicals classified as an eye irritant according to CLP regulation.