N,N,N',N'-tetramethylhexamethylenediamine

Endpoint summary

Description of key information

Studies of skin irritation in the rabbit identified corrosivity and severe eye irritaion. The results of an acute inhalation toxicity study indicate that mortality is associated with local irritation.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Jan 1982

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

(1981-05-12)

GLP compliance:

no

Remarks:

when study was done GLP was not mandatory

Specific details on test material used for the study:

- Name of test material (as cited in study report): N, N, N', N'-Tetramethyl-1,6-hexanediamine
- Analytical purity: 97.5 %
- Expiration date of the lot/batch: stable until 1984-01
- Impurities (identity and concentrations): N,N',N'-Trimethyl-1,6-hexanediamine 2 %

Species:

rabbit

Strain:

Vienna White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Gaukler, Offenbach/Main, Germany
- Housing: single housing in cages made of stainless steel with mire mesh walk floor, floor area: 40x51 cm, no bedding in the cages
- Diet (e.g. ad libitum): Ovator Solikanin 4mm, Muskator-Werke, Düsseldorf, Germany
- Water (e.g. ad libitum): fully demineralized water (monday to friday) and tap water (saturday/ sunday)
- Acclimation period: 8 days before study begin, same housing conditions as during the study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 26
- Humidity (%): 30 - 70
- Air changes (per hr): 15 - 20 times
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

occlusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL (test patch dipped in undiluted substance)

Duration of treatment / exposure:

3 min, 1 hr

Observation period:

72 hrs
reading time points: 30 - 60 min after removal of test patches; 24, 48 and 72 hrs after beginning of the application

Number of animals:

2 male 1 female animal

Details on study design:

TEST SITE
- Area of exposure: 2.5 x 2.5 cm

REMOVAL OF TEST SUBSTANCE
- Washing (if done): at the end of exposure period with Lutrol and Lutrol/water (1:1)
- Time after start of exposure: 3 min, 1 hr

SCORING SYSTEM: Draize scoring system (Draize et al. Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics. In Dermal Toxicity 46-59, 1959. Austin, Tex.: Association of Food and Drug Officials of the United States, Texas, State Department of Health)

Irritation parameter:

other: necrosis

Basis:

mean

Time point:

24/48/72 h

Remarks on result:

other: In experiments with 1 hr exposure time: after 72 hrs deep necrosis in all animals; in experiments with 3 min exposure time: marked necrosis after 72 hrs

Irritation parameter:

erythema score

Basis:

mean

Remarks:

1 hr exposure

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Remarks:

(within 72 hrs)

Irritation parameter:

erythema score

Basis:

mean

Remarks:

3 min exposure

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Remarks:

(within 72 hrs)

Irritation parameter:

edema score

Basis:

mean

Remarks:

1 hr exposure

Time point:

24/48/72 h

Score:

2.7

Max. score:

4

Reversibility:

not reversible

Remarks:

(within 72 hrs)

Remarks on result:

other: extending beyond the area of exposure; finding confirmed by gross-pathological examiniation

Irritation parameter:

edema score

Basis:

mean

Remarks:

3 min exposure

Time point:

24/48/72 h

Score:

1.7

Max. score:

4

Reversibility:

not reversible

Remarks:

(within 72 hrs)

Irritant / corrosive response data:

Necrosis was seen in both experiments (3 min and 1 hr exposure time) in all animals after 24 hrs observation time. After 72 hrs a marked necrosis could be detected for experiments with 3 min exposure time, and a deep necrosis for experiments with 60 min exposure time.
Severe to very severe erythema were seen in all animals over all reading time points (24, 48, 72 hrs) in both experiments (exposure time 1 hr and 3 min). Consequently this effect is not reversible within the given timepoints.
Clearly visible edema were seen in all animals after 24 and 48 hrs in experiments with 3 minutes exposure time. After 72 hrs edema score still very weak edema were seen. Consequently this effects is not reversible within 72 days. The edema extend beyond the exposed area. The edema were confirmed by gross-pathological examinations. In tests with an exposure time of 60 min more severe edema (grade 3 moderate to severe edema) were seen after 24 and 48 hrs in all animals. After 72 hrs the edema were reduced to grade 2 (clearly visible). Again the edema covered an area larger than the exposure size. Within the observation time all edema effects are not reversible.

Animals were sacrificed after 3 days.

Table 1: Skin irritation score for exposure time of 3 and 60 minutes

	3 minutes exposure time			60 minutes exposure time
Readings	Animal	Erythema  score	Edema  score	Animal	Erythema  score	Edema  score
24 hrs	1	4	2	1	4	3
	2	4	2	2	4	3
	3	4	2	3	4	3
48 hrs	1	4	2	1	4	3
	2	4	2	2	4	3
	3	4	2	3	4	3
72 hrs	1	4	1	1	4	2
	2	4	1	2	4	2
	3	4	1	3	4	2
Mean	1	4	1.7	1	4	2.7
per animal	2	4	1.7	2	4	2.7
24, 48, 72 hrs	3	4	1.7	3	4	2.7
Mean (24, 48, 72 hrs)		4	1.7		4	2.7

Interpretation of results:

Category 1A (corrosive) based on GHS criteria

Conclusions:

The substance was shown to be corrosive under the conditions of this study.

Executive summary:

A test was done for assessing the corrosive or irritating potential of the test substance.

The test was done equivalent to OECD guideline 404 (adopted 1981 -05 -12) under occlusive conditions with 2 different exposure times (3 min and 1 hr). Two male and 1 female white vienna rabbits were tested per application .

After 24 hrs (1 st observation time point) necrosis was seen in all animals in both experiments. After 72 hrs a deep necrosis could be seen for experiments with longer exposure time (1 hr) and a marked necrosis for the test with shorter (3 min) exposure time. Severe to very severe erythema were seen in all animals over all reading time points (24, 48, 72 hrs) in both experiments (exposure times 1 hr and 3 min). These effects were not reversible within observation period. Additionally in both experiments edema were seen. For tests with shorter exposure time the edema were of grade 2 for 24 and 48 hrs and decrease to score 1 after 72 hrs. For tests with longer exposure time moderate to severe edema were detected after 24 and 48 hrs. These effects decreased to grade 2 (clearly visible) for tests after 72 hrs. Due to the irreversible damage to the skin, the compound is to be considered as corrosive.

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

7th March 2003 to 22nd March 2003.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

not specified

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

not specified

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Deviations:

not specified

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (MAFF)

Deviations:

not specified

Principles of method if other than guideline:

Not applicable.

GLP compliance:

yes

Specific details on test material used for the study:

- Name of test material (as cited in study report): TOYOCAT-MR
- Physical state: Clear, colourless liquid.
- Analytical purity: not indicated by the sponsor, treated as 100 % pure.
- Lot/batch No.: 1Y0908
- Expiration date of the lot/batch: 12th December 2003
- Storage condition of test material: In the dark at room temperature.
- Other: Specific gravity: 0.80

Species:

rabbit

Strain:

New Zealand White

Remarks:

SPF quality

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Kisslegg, Germany.
- Age at study initiation: At least 6 weeks old.
- Weight at study initiation: At least 1.0 kg.
- Housing: Housed individually in labelled cages with perforated floors (56 x 44 x 37.5 cm).
- Diet (e.g. ad libitum): Approximately 100 g of standard laboratory rabbit diet per day. Hay was also provided twice a week.
- Water (e.g. ad libitum): Tap water provided ad libitum.
- Acclimation period: At least 5 days before the start of treatment under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 3 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): Approximately 15 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours dark.

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

not specified

Amount / concentration applied:

0.5 ml

Duration of treatment / exposure:

3 minutes

Observation period:

2 minutes

Number of animals:

One

Details on study design:

TEST SITE
Approximately 24 hours prior to treatment, the dorsal fur was clipped from the animal, exposing an area approximately 150 square centimetres. Only one test animal was used initially. The test animal was treated with 0.5 ml of the test substance which was aplied to the intact shaved area of one flank using a Metalline patch of 2 x 3 cm. The patch was mounted on Micropore tape which was wrapped around the abdomen and secured with Coban elastic bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated skin was cleaned of residual test substance using water after removal of the dressing.
- Time after start of exposure: The dressing was removed and the treated area were cleaned after the 3 minute exposure period and the 2 minute observation period.

SCORING SYSTEM: Draize scoring system.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 3 minutes

Score:

2

Max. score:

2

Reversibility:

no data

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 3 minutes

Score:

1

Max. score:

1

Reversibility:

no data

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 2 minute observation period

Score:

4

Max. score:

4

Reversibility:

no data

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 2 minute observation period.

Score:

1

Max. score:

1

Reversibility:

no data

Irritant / corrosive response data:

Following three minutes of exposure to 0.5 ml TOYOCAT-MR, well defined erythema and very slight oedema were observed in the treated skin-area of the rabbit immediately after removal of the bandage. Two minutes after this, the degree of skin irritation was had increased to severe erythema / necrosis and very slight oedema.

Other effects:

No symptoms of systemic toxicity were observed in the animals during the test period and no mortalities occurred. Grey discolouration of the treated skin area was observed during the 2 minute observation period.

Due to the corrosivity observed during the 2 minute observation period, no further animals were tested.

Individual Skin Irritation Scores:

	3 minutes treatment site
Time after Exposure	Erythema	Oedema	Comments
Immediately	2	1	-
2 minutes	4	1	k

k = grey discolouration, a sign of necrosis.

Interpretation of results:

Category 1A (corrosive) based on GHS criteria

Conclusions:

Under the conditions of this study, TOYOCAT-MR should be considered to highly corrosive to the skin when tested on New Zealand White rabbit.

Executive summary:

In a study conducted on one male New Zealand White rabbit, the test material, TOYOCAT-MR was applied in a quantity of 0.5 ml to the shaved, intact area of skin on the dorsal side of the rabbit (test-area site approximately 150 square centimetres). Once applied, the test material was covered with a semi-occlusive dressing which remained in place for 3 minutes. After 3 minutes, the dressing was removed and observations were made immediately after removal of the dressing and again, after 2 minutes.

Three minutes exposure to the test material resulted in well-defined erythema and very slight oedema in the treated skin-area of the rabbit immediately after removal of the bandage. Readings after the 2 minute observation period indicated skin irritation had increased to sever erythema/necrosis and very slight oedema. Under the conditions of this study, TOYOCAT-MR should be considered to highly corrosive to the skin when tested on New Zealand White rabbit and shoudl be classified as a Corrosive 1A substance according to CLP Regulation (EC) No. 1272/2008.

Reference 3

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:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

There is no information on the methodology used (such as purity of test, the strain of the animal, the source and age of the animals, on housing and environmental conditions and the site of test site, the method used for assessment etc...).

GLP compliance:

no

Remarks:

not mandatory when the study was carried out

Specific details on test material used for the study:

- Name of test material (as cited in study report): NIAX® Catalyst C-210
- Physical state: yellow, transparent, non-viscous liquid

Species:

rabbit

Strain:

not specified

Details on test animals or test system and environmental conditions:

No information available

Type of coverage:

not specified

Preparation of test site:

not specified

Vehicle:

not specified

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL

Duration of treatment / exposure:

4 hour irritancy test. However, when necrosis was observed shorter test of 1 hour and/or 3 minutes.

Observation period:

Up to 14 days in the 4 hour exposure test and up to 3 days in the 3 minute exposure test.

Number of animals:

6 animals in the 4 hour exposure test and 6 animals in the 3 minute exposure test.

Details on study design:

SCORING SYSTEM: Not specified

Irritation parameter:

other: severe erythema, oedema and necrosis

Reversibility:

not reversible

Remarks on result:

other: These conditions persisted in 6/6 animals until study termination (day 14)

Irritation parameter:

other: severe erythema. oedema and necrosis

Reversibility:

not reversible

Remarks on result:

other: These conditions persisted in 6/6 animals until study termination (day 3).

Irritant / corrosive response data:

4 hour exposure: Severe erythema, oedema and necrosis were reported in all 6 animals and persisted until day 14. Scabs appeared on all animals. Two animals displayed ulceration within 10 days.
3 minute exposure: Severe erythema, oedema and necrosis were reported in all 6 animals and persisted until day 3. Scabs appeared on all animals. Three animals displayed ulceration after 1 or 2 days. Signs of healing of the ulceration was reported on day3.

Interpretation of results:

Category 1A (corrosive) based on GHS criteria

Conclusions:

The substance was shown to be corrosive (category 1A) under the conditions of this study.

Executive summary:

The test substance NIAX® Catalyst C·210 (0.5 mL) was applied to the skin of six rabbits. Animals were exposed to the test substance for 4 hours under an occlusive patch and were restrained during that time. Animals were observed until day 14. Due to corrosivity, a 3 minute exposure tset was also completed on 6 animals and animals were observed for 3 days. Skin reactions were reported to allow classification according to Internation al Air Transport Association (IATA) regulation.

After the 4 hour exposure test, severe erythema, oedema and necrosis was reported in all 6 animals and persisted until day 14. Scabs appeared on all animals. Two animals displayed ulceration within 10 days. Based on these results a 3 minute contact test was carried out to allow for the transport classification (IATA).

After the 3 minute exposure test, severe erythema, oedema and necrosis was reported in all 6 animals and persisted until day 3. Scabs appeared on all animals. Three animals displayed ulceration after 1 or 2 days. Signs of healing of the ulceration was reported on day3.

Due to the irreversible damage to the skin, the substance is considered highly corrosive. Under the conditions of this study,

NIAX® Catalyst C·210 is classified as a Corrosive 1A substance according to Regulation (EC) No 1272/2008.

Reference 4

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Not specified

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:

There is no information on the purity of test, the source and age of the animals, on housing and environmental conditions. However, it is considered that the study still provides scientific and valid information.

GLP compliance:

no

Remarks:

not mandatory when the study was carried out

Specific details on test material used for the study:

- Name of test material (as cited in study report):NIAX Catalyst C-210
- Purity: As supplied
- Physical state: liquid

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

No information available in the test report

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL

Duration of treatment / exposure:

4 hour application

Observation period:

21 days
Reading time points: 1 hour, 1 day, 2, 3, 7, 10, 14 and 21 days

Number of animals:

6 animals (3 males and 3 females)

Details on study design:

SCORING SYSTEM: Draize method

Irritation parameter:

other: necrosis, ecchymosis and scab

Basis:

mean

Time point:

24/48/72 h

Reversibility:

not reversible

Remarks on result:

other: In experiments, necrosis and ecchymosis was reported one hour after patch removal. The formation of scabs appeared at 48 hours and desquamation was reported from day 7 onwards. After 10 dyas, partial peeling of the scabs exposed underlying tissue.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

3.7

Reversibility:

not reversible

Irritation parameter:

erythema score

Basis:

animal #4

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

erythema score

Basis:

animal #5

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

erythema score

Basis:

animal #6

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

3.7

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

3.7

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

edema score

Basis:

animal #4

Time point:

24/48/72 h

Score:

3.7

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

edema score

Basis:

animal #5

Time point:

24/48/72 h

Score:

3.7

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

edema score

Basis:

animal #6

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Irritant / corrosive response data:

Necrosis was seen in all animals at the 1 hour observation time following a 4 hour exposure. Animals showed severe erythema to slight eschar formation in all animals at the 24 and 48 hour timepoints and 5/6 animals at the 72 hour timepoint. One animal showed moderate to severe erythema at the 72 hourtimepoint . At the end of the 21 day observation, the animals did not recover and showed moderate to severe erythema. One animal still displayed severe erythema to slight eschar formation.
Severe odema was noted in all animals at the 24 and 48 hour timepoints and 2/6 animals at the 72 hours timepoint. Moderate to severe erythema was seen in 4/6 animals at the 72 hour timepoint. After 21 days, moderate to severe erythema and severe oedema was still reported in most animals.
Scabs were reported to be peeling at 10 days and showed destruction of the outer layers of tissue. Infection, discharge and severely swollen perimeters were noted at day 21.

Table 1: Skin irritation scores

	Erythema and eschar formation						Oedema
Animal	83-9160	83-9165	83-9283	83-9259	83-9210	83-29214	83-9160	83-9165	83-9283	83-9259	83-9210	83-29214
Readings
24 hours	4	4	4	4	4	4	4	4	4	4	4	4
48 hours	4	4	4	4	4	4	4	4	4	4	4	4
72 hours	4	4	3	4	4	4	3	4	3	3	3	4
7 days	3	3	3	4	3	3	3	3	3	4	3	3
10 days	3	2	3	3	3	3	3	3	3	4	3	3
14 days	2	2	3	3	4	4	2	2	3	3	4	4
21 days	3	3	3	3	4	3	3	3	2	3	4	4
Mean per animal (24-72 hours)	4	4	3.7	4	4	4	3.7	4	3.7	3.7	3.7	4
Mean (24-72 hours)	3.9						3.8

Interpretation of results:

Category 1 (corrosive) based on GHS criteria

Conclusions:

The substance was shown to be corrosive under the conditions of this study. However, it is not possible to distinguish between Category 1A, 1B and 1C.

Executive summary:

The test substance NIAX® Catalyst C·210 (0.5 mL) was applied to the clipped intact area of skin of three males and three females New Zealand White rabbits. Animals were exposed to the test substance for 4 hours under an occlusive patch and were restrained during that time. Excess substance was removed after contact. Skin reactions were reported according to the Draize method at 1 hour, one day, 2, 3, 7, 10, 14 and 21 days.

After 1 hour, 24, 48 and 72 hours, necrosis was reported in all animals. Severe to very severe erythem was seen in all animals at the 24 and 48 hour timepoints. These effects were not reversible within the 21 day observation period. Severe odema was noted in all animals at the 24 and 48 hour timepoints and 2/6 animals at the 72 hours timepoint. Moderate to severe erythema was seen in 4/6 animals at the 72 hour timepoint. After 21 days, moderate to severe erythema and severe oedema was still reported in most animals. Scabs were reported to be peeling at 10 days and showed destruction of the outer layers of tissue. Infection, discharge and severely swollen perimeters were noted at day 21.

Due to the irreversible damage to the skin, the substance is considered highly corrosive. Under the conditions of this study,

NIAX® Catalyst C·210 is classified as a Corrosive 1 (Category 1A, 1B or 1C) substance according to Regulation (EC) No 1272/2008.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (corrosive)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Not specified

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:

There is no information on the purity of test, the source and age of the animals, on housing and environmental conditions. However, it is considered that the study still provides scientific and valid information.

GLP compliance:

no

Remarks:

not mandatory when the study was carried out

Specific details on test material used for the study:

- Name of test material (as cited in study report): NIAX Catalyst C-210
- Physical state: Transparent, pale, yellow, non-viscous liquid
- Purity test date: As supplied

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

No information available in the test report

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.005 mL

Duration of treatment / exposure:

The eyelids were held together for one second following instillation to prevent immediate removal of test material. The eyes were not subsequently rinsed or flushed with water, saline or fluorescein.

Observation period (in vivo):

Up to 21 days. Eye effects were recorded at 1 hour, 4, 24, 48 and 72 hours, 7, 14 and 21 days

Number of animals or in vitro replicates:

6 animals (3 males and 3 females)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): None reported

SCORING SYSTEM: Similar to the scale described in OECD guideline 405.

TOOL USED TO ASSESS SCORE: fluorescein (2 %) staining was used to determine corneal injury before dosing and at readings after one day.

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

3.3

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

cornea opacity score

Basis:

animal #4

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

cornea opacity score

Basis:

animal #5

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

cornea opacity score

Basis:

animal #6

Time point:

24/48/72 h

Score:

3.3

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

not reversible

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

not reversible

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

not reversible

Irritation parameter:

conjunctivae score

Basis:

animal #4

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

not reversible

Irritation parameter:

conjunctivae score

Basis:

animal #5

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

not reversible

Irritation parameter:

conjunctivae score

Basis:

animal #6

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

not reversible

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

3.7

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

chemosis score

Basis:

animal #4

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

conjunctivae score

Basis:

animal #5

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

chemosis score

Basis:

animal #6

Time point:

24/48/72 h

Score:

4

Max. score:

4

Reversibility:

not reversible

Irritant / corrosive response data:

Severe corneal opacity and conjunctival redness and chemosis were reported in all animals. All 6/6 animals at 24 and 48 hours and 4/6 animals at 72 hours displayed opalescent areas, iris details not visible and pupil size barely discernible. Two animals showed corneal opacity at the 72 hour timepoint. On day 7, 4/6 animals showed corneal opacity. By the end of the study all animals were affected. Damage to the cornea did not allow for the examination the iris of each eye.
Diffuse beefy red eyes were seen in all animals throughout the study. Most animals had swelling with lids about half closed to completely closed up to 72 hours after exposure. On day 7, chemosis decreased in one animals but 5/6 animals still showed swelling with lids about half closed to completely closed. On day 14, swelling with lids about half closed remained in 3/6 animals and a decrease in conjunctival chemosis was observed in 3 animals. However at the end of the study swelling with lids about half closed was reported in most animals.
The test substance is highly corrosive and the effects reported 1 hour after exposure persisted until the end of the study. Only chemosis was slightly reduced at the end of the study.

Other effects:

Immediate discomfort was reported by excessive blinking and pawing at eyes. Necrosis on conjunctivae and nictitating membrane along with red brown discharge were noted. At 3 days one rabbit developed a corneal bulge, Corneal bulge abd corneal vascularisation was noted in several animals between day 2 and 14. An opaque white surface on the cornea was also apparent in a few animals.

Table 1: Cornea and Iris irritation scores

	Cornea opacity						Iris
Animal	83-22784	83-22783	83-22782	83-22815	83-22816	83-22820	83-22784	83-22783	83-22782	83-22815	83-22816	83-22820
Readings
1 hour	2	3	3	3	3	2	*	*	*	*	*	*
4 hours	2	3	3	3	3	2	*	*	*	*	*	*
24 hours	3	3	3	3	3	3	*	*	*	*	*	*
48 hours	3	3	3	3	3	3	*	*	*	*	*	*
72 hours	3	3	4	3	3	4	*	*	*	*	*	*
7 days	4	4	4	3	3	4	*	*	*	*	*	*
14 days	4	4	4	4	4	4	*	*	*	*	*	*
21 days	4	4	4	4	4	4	*	*	*	*	*	*
Mean per animal (24-72 hours)	3	3	3.3	3	3	3.3	*	*	*	*	*	*
Mean (24-72 hours)	3.1						*

*Scoring not possible due to cornea opacity

Table 2: Conjunctival redness and chemosis scores

	Conjunctival redness						Chemosis
Animal	83-22784	83-22783	83-22782	83-22815	83-22816	83-22820	83-22784	83-22783	83-22782	83-22815	83-22816	83-22820
Readings
1 hour	3	3	3	3	3	3	3	4	4	3	4	4
4 hour	3	3	3	3	3	3	4	4	4	3	4	4
24 hours	3	3	3	3	3	3	3	4	4	4	3	4
48 hours	3	3	3	3	3	3	4	4	4	4	3	4
72 hours	3	3	3	3	3	3	4	4	4	4	3	4
7 days	3	3	3	3	3	3	4	4	2	3	3	3
14 days	3	3	2	2	2	3	3	3	2	2	2	3
21 days	3	3	2	2	2	3	3	3	3	2	3	3
Mean per animal (24-72 hours)	3	3	3	3	3	3	3.7	4	4	4	3	4
Mean (24-72 hours)	3						3.8

 

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

The substance was shown to cause seious eye damage under the conditions of this study.

Executive summary:

 The test substance NIAX® Catalyst C·210 (0.005 mL) was instilled into the conjunctival sac of one eye of each animal (three males and three females New Zealand White rabbits). The eyelids was held together for one second. The eyes were examined and scored at one hour, 4, 24, 48 and 72 hours, 7, 14 and 21 days. Fluorescein was used to determine corneal injury before dosing and at readings after 24 hours.

Corneal opacity and conjunctival injury was reported in all animals and persisted through the study. Due to corneal opacity, the iris could not be examined. Necrosis of conjunctivae and nictitating membrane were observed. Corneal bulge and corneal vascularisation was apparent in several animals between day 2 and 14.

Under the conditions of this study, NIAX® Catalyst C·210 is classified as serious damage eye damage 1 ( substance according to Regulation (EC) No 1272/2008.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Additional information

The substance is shown to be corrosive to skin and a severe eye irritant; classification as Corrosive is proposed, according to CLP criteria.

Justification for classification or non-classification

Based on the available data, the substance is classified according to the CLP Regulation (Reulation 1272/2008) as Corrosive (Category 1A) with the hazard statement H314: Causes severe skin burns and eye damage.