Registration Dossier

Administrative data

Description of key information

Skin irritation (equivalent to OECD 404), rabbit: not irritating

Eye irritation (equivalent to OECD 405), rabbit: serious eye damage

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:
key study
Study period:
7-10 Oct 1997
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Version / remarks:
not specified
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Version / remarks:
2015
Deviations:
yes
Remarks:
Occlusive dressing instead of semi-occlusive dressing was used.
GLP compliance:
yes
Species:
rabbit
Strain:
other: Hra:(NZW)SPF
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Covance Research Products lnc. (Denver, PA)
- Age at study initiation: at least three months old
- Weight at study initiation: at least 2000 g
- Housing: individually housed in suspended, stainless-steel mesh cages in an American Association for Accreditation of Laboratory Animal Care-accredited vivarium
- Diet: certified High Fiber Rabbit Diet (PMI® #5325) available ad libitum
- Water: local public water system available ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-21
- Humidity (%): 60-68
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
occlusive
Preparation of test site:
shaved
Vehicle:
water
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied: 0.5 g moistened with water
Duration of treatment / exposure:
4 h
Observation period:
The site of application was examined at 1, 24, 48, and 72 hours after removal of the occlusive patch.
Number of animals:
3
Details on study design:
TEST SITE
- Area of exposure: dorsal skin
- % coverage: not reported
- Type of wrap if used: occlusive

REMOVAL OF TEST SUBSTANCE
- Washing: yes
- Time after start of exposure: 4 h after exposure

OBSERVATION TIME POINTS
The site of application was examined at 1, 24, 48, and 72 hours after removal of the occlusive patch.

SCORING SYSTEM:
- Method of calculation: Graded as described in OECD Guideline 404 (Annex V., Test B.4) (Grading of Skin Reaction)
Irritation parameter:
erythema score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Irritation parameter:
edema score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Interpretation of results:
other: CLP/EU GHS criteria are not met, no classification required according to Regulations (EC) No 1272/2008.
Conclusions:
In the reliable skin irritation study performed with N-(2-(( 4-Amino-3-methylphenyl)ethylamino) ethyl)methanesulfonamide sulfate (2:3) monohydrate (CAS 25646-71-3) according to OECD TG 404 three albino rabbits were exposed to 0.5 g of the neat test material for 4 h applied onto the shaved skin via occlusive dressing. No edema formation was observed in any animal. The mean values for erythema and edema were calculated to be 0, respectively. Therefore, the test substance does not need to be classified for skin irritation according to Regulation (EC) No 1272/2008.
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:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
not specified
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
adopted 9 October 2017
Deviations:
yes
Remarks:
Deviations: only 2 animals instead of three; in one animal the eye was washed out immediately after treatment (TG: after 24 hrs)
GLP compliance:
yes
Species:
rabbit
Strain:
other: Hra:(NZW)SPF
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Covance Research Products lnc. (Denver, PA)
- Age at study initiation: at least three months old
- Weight at study initiation: at least 2000 g
- Housing: individually housed in suspended, stainless-steel mesh cages in an American Association for Accreditation of Laboratory Animal Care-accredited vivarium
- Diet: certified High Fiber Rabbit Diet (PMI® #5325) available ad libitum
- Water: local public water system available ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-22
- Humidity (%): 46-62
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied: 0.1 g
Duration of treatment / exposure:
One of the treated eyes was immediately washed with running distilled water
Observation period (in vivo):
Eyes were observed immediately after instillation of the test substance and 1, 24, 48, and 72 hours, and 7, 14, 21, and 28 days thereafter.
Number of animals or in vitro replicates:
2
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing: yes
- Time after start of exposure: One of the treated eyes were immediately washed with running distilled water.

SCORING SYSTEM: Graded as described in OECD Guideline 405 (Annex V., Test B.5) (Grading for Ocular Lesions)

TOOL USED TO ASSESS SCORE: fluorescein
Irritation parameter:
cornea opacity score
Remarks:
unwashed eyes
Basis:
mean
Time point:
24/48/72 h
Score:
0.33
Max. score:
4
Reversibility:
fully reversible within: 48 h
Irritation parameter:
iris score
Remarks:
unwashed eyes
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: not applicable
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
24/48/72 h
Score:
2.33
Max. score:
3
Reversibility:
fully reversible within: 28 days
Irritation parameter:
chemosis score
Remarks:
unwashed eyes
Basis:
mean
Time point:
24/48/72 h
Score:
3
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritant / corrosive response data:
For the unwashed eye, chemosis (grade 4) for the conjuctiva and a profuse discharge was noted one hour after administration of the test substance.
Signs of irritation for the unwashed eye between the 24 and 72 hour examination included corneal opacity (grade 1), redness of the conjunctiva (grade 3) and chemosis (grade 3) of the nictitating membrane/or lids. In addition, staining of the cornea and conjunctiva were apparent for the unwashed eye when fluorescein dye was applied at the 24-hour examination. Since the test substance caused significant ocular lesion within 72 hours after exposure which persisted for more than 24 hours, these changes were considered sufficient to characterize the response as irritating to eyes.
Other effects:
- Lesions and clinical observations: Animals were not weighed as weights are not critical to this study.
- Ophthalmoscopic findings: None reported
- Histopathological findings: No necropsies were conducted at the conclusion of the observation period.
- Effects of rinsing or washing: Immediate irrigation of the eye was palliative. For the washed eye, signs of irritation were limited to redness (grade 2) and chemosis (grade 2). A profuse discharge was also seen one hour after administration of the test substance. No staining was evident when fluorescein dye was applied. By Day 7 of the study, the washed eye appeared normal.
Interpretation of results:
other: CLP/EU GHS Category 1 (H318) according to Regulation (EC) No 1272/2008
Conclusions:
In an in vivo eye irritation study (OECD 405), significant irritation was observed that was not reversible until day 21 of the study. The available data on eye irritation of the substance meet the criteria for classification as serious eye damage/eye irritation Cat. 1 (H318) according to Regulation (EC) 1272/2008.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

In a GLP compliant, reliable skin irritation study (according to OECD 404) performed with N-(2-(4-Amino-N-ethyl-m-toluidino)ethyl)methanesulphonamide sesquisulphate, 3 rabbits were exposed to 0.5 g of the diluted test material for 4 h applied onto the shaved skin via occlusive dressing (Shepard, 1997). Skin reactions were evaluated according to the Draize scoring system 1, 24, 48 and 72 h post-application. No symptoms of systemic toxicity were observed in any animal during the test period and no mortality occurred during the course of the study. Mean erythema and edema scores for the 72 hour observation period were 0, respectively. Therefore, the test substance does not need to be classified for skin irritation according to Regulation (EC) No 1272/2008.

 

A GLP compliant, reliable eye irritation study (according to OECD 405) with restrictions (only two animals tested, in one animal the eye was washed out immediately after instillation of the test item) was performed with N-(2-(4-Amino-N-ethyl-m-toluidino)ethyl)methanesulphonamide sesquisulphate (Shepard, 1998). In the study 0.1 mL of the diluted test material was instilled into the eyes of 2 rabbits. The eyes were examined and the changes were graded according to the Draize scoring system 24, 48 and 72 h post-application and for a prolonged observation period of 28 days. No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred during the course of the study.For the unwashed eye, chemosis (grade 4) for the conjuctiva and a profuse discharge was noted one hour after administration of the test substance.

Signs of irritation for the unwashed eye between the 24 and 72 hour examination included corneal opacity (grade 1), redness of the conjunctiva  redness (grade 3) and chemosis (grade 3) of the nictitating membrane/or lids. In addition, staining of the cornea and conjunctiva were apparent for the unwashed eye when fluorescein dye was applied at the 24-hour examination. These changes were sufficient to characterize the response as significant irritation. Thus, the mean values for corneal opacity, iris and conjunctival irritation and chemosis were 0.33, 0, 2.33, and 3.00, respectively. At the end of the observation period as recommended in OECD TG 405 of 21 days, conjunctival redness grade 2 was still apparent in the unwashed eye. Therefore the observation period was prolonged up to 28 days after which all signs of eye irritation had reversed. However, since only one eye was available for assessment of effects on the eye and the observed effects were not reversible within the recommended observation period, irreversible effects on the eyes cannot be excluded. Thus, the test item should be classified as causing serious eye damage.

Justification for classification or non-classification

The available data on skin irritation of N-(2-(4-Amino-N-ethyl-m-toluidino)ethyl)methanesulphonamide sesquisulphate do not meet the criteria for classification according to Regulation (EC) No 1272/2008, and are therefore conclusive but not sufficient for classification.

N-(2-(4-Amino-N-ethyl-m-toluidino)ethyl)methanesulphonamide sesquisulphate is classified as serious eye damage/eye irritation category 1 with the hazard statement H318 ‘Causes serious eye damage.’ according to Regulation (EC) 1272/2008.