Reaction mass of 1H-Benzotriazole-1-methanamine, N,N-bis(2-ethylhexyl)-6-methyl- and 2H-Benzotriazole-2-methanamine, N,N-bis(2-ethylhexyl)-5-methyl- and N,N-bis(2-ethylhexyl)-4-methyl-1H-benzotriazole-1-methylamine and 2H-Benzotriazole-2-methanamine, N,N-bis(2-ethylhexyl)-4-methyl- and N,N-bis(2-ethylhexyl)-5-methyl-1H-benzotriazole-1-methylamine

Endpoint summary

Description of key information

The test substance caused skin irritation on rabbit skin. Erythema and edema with a score of 2 - 3 were evident in all 6 animals. Only very slight effects in the rabbit eye were caused by the substance.

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:

August 3, 1981 - August 10, 1981

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

(limited documentation, occlusive treatment, 24 h exposure)

Qualifier:

according to guideline

Guideline:

other: (Proposed Guidelines of the United States Environmental Agency (EPA), §163.81-5 "Primary dermal irritation study", Federal Register, Vol. 43, No. 163, August 22, 1978).

Principles of method if other than guideline:

Gauze patches of 2.5 x 2.5 cm laden with 0.5 mL of the test material were applied to the prepared abdraded and intact skin for 24 hours. Scoring was recorded 24 h, 48 h, 72 h, 4 d and 7 d after application.

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: bred and raised in Ciba-Geigy premises
- Age at study initiation: adult
- Weight at study initiation: 2-3 kg
- Housing: housed individually in metal cages
- Diet: standard rabbit food (NAFAG, No. 814, Gossau SG); ad libitum
- Water: ad libitum
- Acclimation period: minimum of 4 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 55 ± 10%
- Photoperiod (hrs dark / hrs light): 12/ 12

Type of coverage:

occlusive

Preparation of test site:

other: abraded and intact skin

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 mL of test material on gauze patches of 2.5 x 2.5 cm

Duration of treatment / exposure:

24h

Observation period:

7 days

Number of animals:

6 (3 males and 3 females)

Details on study design:

TEST SITE
- Site of exposure: back and flank of the rabbits
- Area of exposure: 2.5 x 2.5 cm²
- Type of wrap if used: gauze patches covered with an impermeable material, fastened to the body of the rabbits with adhesive tape

REMOVAL OF TEST SUBSTANCE
- Time after start of exposure: 24h

SCORING SYSTEM:
- Evaluation was performed according to the Draize Scoring System.
- Calculation of the primary skin irritation index: the scores read after 24 and 72 hours for erythema and edema for the intact as well as for the abraded skin were summed up and divided by 4.
- Assessment of irritation:
0
0.0-1.0 : minimal
1.1-2.0 : slight
2.1-4.0 : moderate
4.1-6.0 : marked
6.1-8.0 : extreme

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 days

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

3.33

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 days

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2.67

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 days

Irritation parameter:

erythema score

Basis:

animal #4

Time point:

24/48/72 h

Score:

2.67

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 days

Irritation parameter:

erythema score

Basis:

animal #5

Time point:

24/48/72 h

Score:

2.33

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 days

Irritation parameter:

erythema score

Basis:

animal #6

Time point:

24/48/72 h

Score:

2.33

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 days

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 days

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 days

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 days

Irritation parameter:

edema score

Basis:

animal #4

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 days

Irritation parameter:

edema score

Basis:

animal #5

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 days

Irritation parameter:

edema score

Basis:

animal #6

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 days

Results of Skin irritation (intact skin, 24 h exposure):

Observation time	Rabbit no.
	1		2		3		4		5		6
	Ery.	Edema	Ery.	Edema	Ery.	Edema	Ery.	Edema	Ery.	Edema	Ery.	Edema
24 h	2	3	2	3	2	3	2	3	2	1	2	1
48 h	2	3	4	3	3	3	2	3	2	2	2	1
72 h	2	3	4	3	3	3	4	3	3	2	3	1
4 d	2	3	4	3	3	2	4	2	2	2	3	2
7 d	2	3	3	3	3	2	3	3	3	2	3	2
Mean value 24 + 48 + 72 h	2.00	3.00	3.33	3.00	2.67	3.00	2.67	3.00	2.33	1.67	2.33	1.00
Mean value 24 + 48 + 72 h all animals	2.55	2.44	-	-	-	-	-	-	-	-	-	-

Ery.: Erythema

 

Mean reaction scores:

	Mean reaction score
Time after exposure (hours)	Erythema		Edema		Total
	Intact skin	Abraded skin	Intact skin	Abraded skin
24	2.0	2.3	2.3	2.7
72	3.2	3.3	2.5	2.7
Total	5.2	5.6	4.8	5.8	21

Primary irritation index = 21.0 / 4 = 5.3

The test substance caused a marked irritation when applied to intact and abraded rabbit skin.

Up to 4 days after application, the surface of the erythema was greater than the site of application.

Slightly ischaemic areas were observed after 48 hours in 4/12 and after 3 and 4 days in 10/12 application sites.

After 7 days, practically all animals had cracked skin.

Both erythema and edema were observed with an average score between 2 and 3 (24 to 72h). The skin condition did not improve on day 4 and only barely improved on day 7, which was the last observation time point. The results for abraded skin were not considered for classification.

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Conclusions:

The test article was found to cause a marked irritation when applied to intact and abraded rabbit skin.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

August 3, 1981 - August 10, 1981

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

(limited documentation)

GLP compliance:

no

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Lot/batch No.of test material: PP 126/1

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 2 - 3 kg
- Housing:individually in metal cages, numbered by ear tags
- Diet: NAFAG, No. 814, Gossau SG, ad libitum
- Water: ad libitum
- Acclimation period: minimum of 4 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2
- Humidity (%): 55 ± 10
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:

unchanged (no vehicle)

Controls:

other: untreated right eye served as a control

Amount / concentration applied:

0.1 mL

Duration of treatment / exposure:

one single application

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

6

Details on study design:

SCORING SYSTEM:
Evaluation of the eye reactions according to the Draize Scoring System.

TOOL USED TO ASSESS SCORE: hand-slit lamp

Eyes of 3/6 animals were rinsed after approx. 30 seconds.

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

out of 6 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

other: 3/6 rinsed

Irritation parameter:

iris score

Basis:

mean

Remarks:

out of 6 animals

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Remarks on result:

other: 3/6 rinsed

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

fully reversible within: 72 h

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

not fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.33

Max. score:

3

Reversibility:

not fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Basis:

animal #4

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: rinsed

Irritation parameter:

conjunctivae score

Basis:

animal #5

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: rinsed

Irritation parameter:

conjunctivae score

Basis:

animal #6

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: rinsed

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

animal #4

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: rinsed

Irritation parameter:

chemosis score

Basis:

animal #5

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: rinsed

Irritation parameter:

chemosis score

Basis:

animal #6

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: rinsed

Results: Eye irritation

	Eye treatment	not rinsed			rinsed (after 30 s)
	Animal no.	1	2	3	4	5	6
24 h	Cornea	0	0	0	0	0	0
	Area	0	0	0	0	0	0
	Iris	0	0	0	0	0	0
	Conjunctiva	1	1	2	1	1	1
	Chemosis	0	1	1	1	1	1
	Discharge	0	0	0	0	0	0
48 h	Cornea	0	0	0	0	0	0
	Area	0	0	0	0	0	0
	Iris	0	0	0	0	0	0
	Conjunctiva	1	0	1	1	0	0
	Chemosis	0	0	1	1	1	0
	Discharge	0	0	0	0	0	0
72 h	Cornea	0	0	0	0	0	0
	Area	0	0	0	0	0	0
	Iris	0	0	0	0	0	0
	Conjunctiva	0	1	1	1	0	0
	Chemosis	0	0	1	1	1	0
	Discharge	0	0	0	0	0	0
4 d	Cornea	0	0	0	0	0	0
	Area	0	0	0	0	0	0
	Iris	0	0	0	0	0	0
	Conjunctiva	0	1	1	1	1	0
	Chemosis	0	0	1	1	1	0
	Discharge	0	0	0	0	0	0
7d	Cornea	0	0	0	0	0	0
	Area	0	0	0	0	0	0
	Iris	0	0	0	0	0	0
	Conjunctiva	0	1	1	0	0	1
	Chemosis	0	0	0	0	1	1
	Discharge	0	0	0	0	0	0
Mean value 24, 48, 72 h	Cornea	0	0	0	0	0	0
	Iris	0	0	0	0	0	0
	Conjunctiva	0.67	0.67	1.33	1.00	0.33	0.33
	Chemosis	0	0.33	1.00	1.00	1.00	0.33

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this experiment the test material was found to cause a minimal irritation below the level of regulatory significance when applied to the rabbit eye mucosa.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin Irritation

In a primary dermal irritation study, performed using a method identical to that described in the Federal Register, Vol. 43, No. 163, August 22, 1978, 0.5 mL of the test substance (no data on purity) was applied under occlusive conditions to the intact and abraded skin of 6 Albino rabbits. Treatment was terminated 24 hours following application and the animals were observed for 7 days. Irritation was scored according to the Draize method 24, 48 and 72 hours after application. 5/6 animals had an average erythema score (24 h–72 h) ≥ 2.3 (maximal score = 4). 4/6 animals had an average edema score (24 h–72 h) ≥ 3 (maximal score = 4). None of the observed effects was reversible within the observation period in any animal. Results from the abraded skin are not included in the final judgment for skin irritation of the test substance because of its lack of scientific justification and gross deviation from today's acceptable standardized procedures for the appraisal of skin irritation. In this study, the test substance is a skin irritant. This study is suitable for assessment of dermal irritation as it was performed using a protocol which is similar and equivalent to the OECD guideline. The conditions are even harsher (occlusive, extended exposure) than that described by the actual OECD guideline.

 

Eye irritation

The eye irritation potential of the test substance (no data on purity) was determined in a procedure similar and equivalent to the Test Guideline OECD 405. 0.1 mL of the test substance was applied to the conjunctival sac of the left eye of each of 6 Albino rabbits. The other eye remained untreated and served as control. Irritation was recorded after 24, 48 and 72 hours. Appraisal of eye irritation was performed according to the method of Draize. Average Draize scores (24, 48 and 72 hours) were calculated per animal for corneal opacity, iritis, conjunctivae redness and chemosis. Average values (24 h – 72 h) for cornea and iris score were 0 (maximal score = 4 and 2, respectively) in all animals. Effects on conjunctivae redness were seen in all animals. The average scores were between 0.33 (2/6) and 1.33 (1/6) (maximal score = 3) and were not fully reversible within the observation period in 3/6 animals. The average values for chemosis were between 0 (1/6) and 1 (3/6) and were not fully reversible within 7 days in 2/6 animals. These values are beneath the threshold for classification, hence the test substance is not an eye irritant in this study. This study is suitable for assessment of eye irritation as it was performed using a protocol which is similar and equivalent to the OECD guideline.

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. As a result the substance is considered to be classified and labelled as skin irritant category 2 (H315: "causes skin irritation") but not as eye irritant under Regulation (EC) No 1272/2008, as amended for the eighth time in Regulation (EU) No 2016/218.