Registration Dossier

Administrative data

Description of key information

Skin irritation: The skin irritation potential of the substance was determined with a K1 GLP-study in New Zealand White rabbits according to OECD Guideline 404 and EU Method B.4. The substance was observed not to be irritating to the skin. This study was selected as key study.
Eye irritation: The eye irritation potential of the substance was determined with a K1 study in New Zealand White rabbits according to OECD Guideline 405 and EU Method B.5. The substance was observed to be irritating to the eyes and therefore classified as category 2 according to CLP regulation. This study was selected as key study. In addition, the in vitro potential for eye irritation was assessed with a Bovine Corneal Opacity and Permeability Test, following a protocol equivalent to the OECD Guideline n°437. According to this study, T002102 is not considered as an eye irritant.

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:
2001-09-11 to 2001-09-20
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test material information:
Composition 1
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: RT002102G4A311
- Expiration date of the lot/batch: 2002-03-25 (retest date)
- Purity test date: no data
- Purity: 99.4%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: in the original container, at room temperature (range 17-23°C), away from direct sunlight.
- Stability under test conditions: stable under storage conditions
- Solubility and stability of the test substance in the solvent/vehicle: not applicable
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: not applicable

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: moistened with bi-distilled water before application

Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Elevage Scientifique des Dombes, F-01400 Chatillon sur Chalaronne/France
- Age at start of treatment: 10-11 weeks (male and females)
- Weight at day of treatment: 2257 grams (male), 2113 to 2275 grams (females)
- Housing: Individually in stainless steel cages equipped with feed hoppers, drinking water bowls, with wood and haysticks for gnawing.
- Diet (e.g. ad libitum): pelleted standard Provimi Kliba 3418 rabbit maintenance diet, ad libitum
- Water (e.g. ad libitum): community tap water from Fűllinsdorf, ad libitum
- Acclimation period: 2001-09-11 to 2001-09-16, under laboratory conditions after health examination

ENVIRONMENTAL CONDITIONS
- Temperature (deg C): 17-23 °C
- Humidity (%): 30-70%
- Air changes (per hr): approximately 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark, music was played during the light period

IN-LIFE DATES: From: 2001-09-17 To: 2001-09-20
Type of coverage:
semiocclusive
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 g/animal, weighed as delivered and then moistened with bi-distilled water before appliction.
- Concentration (if solution): not applicable

Duration of treatment / exposure:
4 hours
Observation period:
72 hours after treatment
Number of animals:
3 animals (one male and 2 females)
Details on study design:
TEST SITE
- Area of exposure: flank of the animal. On the day of treatment, 0.5 g of the test substance was placed on a surgical gauze patch (2.5 cm x 2.5 cm). This patch was applied to approximately 6 cm2 of the intact skin of the clipped area.
- % coverage: no data
- Type of wrap if used: semi-occlusive dressing. Dressing was wrapped around the abdomen and anchored with tape

REMOVAL OF TEST SUBSTANCE
- Washing (if done): After treatment, the dressing was removed and the skin was flushed with lukewarm tap water to clean the application site.
- Time after start of exposure: 4 hours

OBSERVATION TIME POINTS: 24, 48 and 72 hours after patch removal

SCORING SYSTEM:
- The skin reaction was assessed according to the numerical scoring system listed in the EEC Commission Directive 92/69/EEC, July 31, 1991.
Irritation parameter:
erythema score
Basis:
mean
Remarks:
male 96
Time point:
24/48/72 h
Score:
0.33
Max. score:
4
Reversibility:
fully reversible within: 48 hours
Remarks on result:
no indication of irritation
Irritation parameter:
erythema score
Basis:
mean
Remarks:
female 97
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
erythema score
Basis:
mean
Remarks:
female 98
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
mean
Remarks:
male 96
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
mean
Remarks:
female 97
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
mean
Remarks:
female 98
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
The mean for erythema/eschar score of the three animals was 0.33, 0.00, and 0.00, respectively and the mean oedema score was 0.00 for all three animals.
Very slight erythema was observed at the test site of one animal from 1 to 24 hours after treatment.
No abnormal findings were observed at the test site of any animal at either the 48- or 72-hour readings.
No irreversible alterations of the treated skin were observed, nor were corrosive effects evident on the skin.
Other effects:
No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.
No staining by the test substance of the treated skin was observed.
The body weights of all rabbits were considered to be within the normal range of variability.

Animal number

Sex

Evaluation Interval

Erythema

Oedema

Scaling

96

M

1 hour

1

0

0

97

F

1 hour

0

0

0

98

F

1 hour

0

0

0

96

M

24 hour

1

0

0

97

F

24 hour

0

0

0

98

F

24 hour

0

0

0

96

M

48 hour

0

0

0

97

F

48 hour

0

0

0

98

F

48 hour

0

0

0

96

M

72 hour

0

0

0

97

F

72 hour

0

0

0

98

F

72 hour

0

0

0

 

Interpretation of results:
GHS criteria not met
Conclusions:
The test substance was considered to be "not irritating" to rabbit skin under the conditions of this study and therefore not to be classified according to CLP regulation.
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
Study period:
2001-09-25 to 2001-10-18
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
Adopted 1987-02-24
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Version / remarks:
Adopted 1992-07-31
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test material information:
Composition 1
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: RT002102G4A311
- Expiration date of the lot/batch: 2002-03-25 (retest date)
- Purity test date: no data
- Purity: 99.4%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: in the original container, at room temperature (range 17-23°C), away from direct sunlight.
- Stability under test conditions: stable under storage conditions
- Solubility and stability of the test substance in the solvent/vehicle: not applicable
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: not applicable

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: none
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Elevage Scientifique des Dombes, F-01400 Chatillon sur Chalaronne, France
- Age at start of treatment: 14-15 weeks (males); 15-16 weeks (females)
- Weight at day of treatment: 2872 g (male), 2705-2736 g (females)
- Housing: Individually in stainless steel cages equipped with feed hoppers, drinking water bowls, with wood and haysticks for gnawing.
- Diet (e.g. ad libitum): ad libitum, pelleted standard Provimi Kliba 3418 rabbit maintenance diet (batch no. 92/01)
- Water (e.g. ad libitum): ad libitum, community tap water from Füllinsdorf
- Acclimation period: 2001-09-25 to 2001-09-30, under laboratory conditions after health examination

ENVIRONMENTAL CONDITIONS
- Temperature (deg C): 20+-3 °C
- Humidity (%): 30-70%
- Air changes (per hr): 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hrs dark / 12 hrs fluorescent light, music played during the light period

IN-LIFE DATES: From 2001-10-01 To 2001-10-18
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied: 0.1 g/animal (left eye only)
- Concentration: no data
Duration of treatment / exposure:
single application, the treated eyes were not rinsed after instillation.
Observation period (in vivo):
17 days
Scoring: 1, 24, 48, and 72 hours, as well as 7, 10, 14 and 17 days after administration
Number of animals or in vitro replicates:
3 animals (1 male; 2 females)
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): no. The treated eyes were not rinsed after instillation.
- Time after start of exposure: not applicable.

SCORING SYSTEM:
The ocular reaction was assessed according to the numerical scoring system listed in the EEC Commission directive 92/69/EEC at approximately 1, 24, 48 and 72 h, as well as 7, 14 and 21 days after administration. When present, corrosion of and/or staining of sclera and cornea by the test article were recorded and reported. Eye examinations were made with Varta Cliptrix diagnostic lamp (A. Riegger, Basel, Switzerland).
The primary irritation score was calculated by totaling the mean cumulative scores at 24, 48 and 72 hours and then divided by the number of data points.
Irritation parameter:
overall irritation score
Remarks:
primary eye irritation
Basis:
mean
Remarks:
cumulative scores at 24, 48, and 72 hours and then divided by the number of data points.
Time point:
24/48/72 h
Score:
3
Reversibility:
fully reversible within: 17 days
Irritation parameter:
cornea opacity score
Remarks:
opacity
Basis:
mean
Remarks:
male 10
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Irritation parameter:
cornea opacity score
Remarks:
opacity
Basis:
mean
Remarks:
female 11
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
cornea opacity score
Remarks:
opacity
Basis:
mean
Remarks:
female 12
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Irritation parameter:
cornea opacity score
Remarks:
area
Basis:
mean
Remarks:
male 10
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Irritation parameter:
cornea opacity score
Remarks:
area
Basis:
mean
Remarks:
female 11
Time point:
24/48/72 h
Score:
3.33
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
cornea opacity score
Remarks:
area
Basis:
mean
Remarks:
female 12
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Irritation parameter:
iris score
Basis:
mean
Remarks:
male 10
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: not applicable
Irritation parameter:
iris score
Basis:
mean
Remarks:
female 11
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: not applicable
Irritation parameter:
iris score
Basis:
mean
Remarks:
female 12
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: not applicable
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Remarks:
male 10
Time point:
24/48/72 h
Score:
2
Max. score:
3
Reversibility:
fully reversible within: 14 days
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Remarks:
female 11
Time point:
24/48/72 h
Score:
2
Max. score:
3
Reversibility:
fully reversible within: 14 days
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Remarks:
female 12
Time point:
24/48/72 h
Score:
1.67
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
male 10
Time point:
24/48/72 h
Score:
0.67
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
female 11
Time point:
24/48/72 h
Score:
1.33
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
female 12
Time point:
24/48/72 h
Score:
0.33
Max. score:
4
Reversibility:
fully reversible within: 48 hours
Irritant / corrosive response data:
- primary irritation score of 3.0
- eye reaction (mean values from 24 to 72 h) consisted:
cornea opacity 0.33
iris lesions 0.0
redness of the conjunctivae 1.89
chemosis of the conjunctivae 0.85.
- A slight corneal opacity, effecting up to the whole area of the cornea, was observed in one animal from 1 to 48 hours after treatment. This opacity was still present at the 72-hour reading (effecting up to half of the cornea) but was no longer apparent 7 days after treatment.
-No abnormal findings were observed in the iris of any animal at any reading during the observation period.
-Slight to moderate reddening of the conjunctivae was noted in two animals from 1 hour to 10 days after treatment and in the other animal from 1 to 72 hours after treatment.
-Swelling of the conjunctivae, with lids about half closed, was evident in all animals at the 1-hour reading. The severity of the swelling decreased at the 24-hour reading and gradually diminished to clear in all animals by the 7-day reading.
-A slight to moderate watery discharge was observed in all animals 1 hour after treatment and persisted in one animal up to the 24-hour reading.
-Assessment of the sclera was not possible 1 hour after treatment due to swelling of the conjunctivae, however, moderate reddening was evident in all animals at the 24-hour reading. Slight to moderate reddening persisted in one animal up to the 72-hour reading, up to the 7 -day reading in another animal and up to the 14-day reading in the remaining animal.
-No abnormal findings were observed in the treated eye of any animal at the final 17-day reading.
-No corrosion of the cornea was observed at any of the reading times. No abnormal effects were observed in the treated eyes of any animal 17 days post-exposure.
Other effects:
- No mortality occurred.
- No clinical signs of systemic toxicity
- No staining from the test substance was observed.
- No remnants of the test item in the eye or conjunctival sac were observed.
- Animal number 10 showed a very slight body weight loss (1 %) during the acclimatization period. This weight loss is considered to be incidental. The body weights of all other rabbits were considered to be within the normal range of variability.

Table 1. Eye irritation scores-mean values after 24, 48 and 72 hours.

Animal Number

Sex

Corneal Opacity

N

Iris

N

Conjunctivae

Primary Eye Irritation Score

Redness

N

chemosis

N

10

M

0.00

3

0.00

3

2.00

3

0.67

3

3.00

11

F

1.00

3

0.00

3

2.00

3

1.33

3

12

F

0.00

3

0.00

3

1.67

3

0.33

3

Mean Score

0.33

0.00

1.89

0.78

 

 N= number of available data points

Interpretation of results:
Category 2 (irritating to eyes) based on GHS criteria
Conclusions:
The test substance is considered to be "not irritating" to the rabbit eye based on the referred classification criteria (EEC Commission Directive 93/21/EEC of April 27, 1993). However, based on the results of this study and the criteria of the CLP Regulation, the substance should be classified as eye irritant category 2.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation / corrosion:

Skin irritation/corrosion potential of the substance was assessed by applying 0.5 g of the substance to an exposure area of 6 cm² of 3 New Zealand White rabbits (two females, one male, semi-occlusive patch). The test site was shaved. Four hours after start of exposure, the test sites were washed with water. Each test site was examined and scored separately for erythema and edema on a graded scale of 0 to 4 (primary dermal irritation index). The mean score was calculated across 3 scoring times (24, 48, and 72 hours after patch removal) for each animal for erythema/eschar grades and for oedema grades, separately. The mean for erythema/eschar score of the three animals was 0.33, 0.00, and 0.00, respectively and the mean oedema score was 0.00 for all three animals.

Very slight erythema was observed at the test site of one animal from 1 to 24 hours after treatment. No abnormal findings were observed at the test site of any animal at either the 48- or 72-hour readings. No irreversible alterations of the treated skin were observed, nor were corrosive effects evident on the skin.

Eye irritation:

One in vivo key study (RCC, 2001) and one in vitro supporting study (F. Van Goethem, 1997) are available for this endpoint.

Eye irritation potential of the substance was assessed in vivo by applying 0.1 g of the substance to the left eye of 3 New Zealand White rabbits (one male and two females). The treated eyes were not rinsed after instillation. The ocular reaction was assessed according to the numerical scoring system listed in the EEC Commission directive 92/69/EEC at approximately 1, 24, 48 and 72 h, as well as 7, 14 and 21 days after administration. Application of the test substance to healthy rabbit conjunctivae resulted in a primary irritation score of 3.0. The eye reaction (mean values from 24 to 72 h) consisted of grade 0.33 cornea opacity, grade 0.0 iris lesions, grade 1.89 redness of the conjunctivae and grade 0.78 chemosis of the conjunctivae. A slight corneal opacity, effecting up to the whole area of the cornea, was observed in one animal from 1 to 48 hours after treatment. This opacity was still present at the 72-hour reading (effecting up to half of the cornea) but was no longer apparent 7 days after treatment. No corrosion of the cornea was observed at any of the reading times. No abnormal effects were observed in the treated eyes of any animal 17 days post-exposure.

The in vitro bovine corneal opacity-permeability (BCOP) assay assessed the ocular irritation potential of T002102 by the use of isolated bovine corneas. Both opacity and permeability were measured and the resulting objective values were combined to calculate the ‘in vitro score’. When tested as a 20 % (g/g) suspension, T002102 induced no increase in opacity (-1.4 ± 0.6) nor in permeability (-0.012 ±0.007). Consequently, an in vitro score of -1.6 ± 0.7 was calculated which classified T002102 as a non-eye irritant. When the test compound was used pure (100%) in the repeat study, again no increase in opacity (0.3 ± 1.0) and in permeability (-0.002 ±0.004) was obtained resulting in an in vitro classificaton (0.3 ± 1.0) of non-irritancy.

The test substance is considered to be "not irritating" to the rabbit eye based on the referred classification criteria (EEC Commission Directive 93/21/EEC of April 27, 1993) and based on the results of the in vitro study. However, according to the criteria of the CLP Regulation the substance is classified as eye irritant category 2 based on the fact that the in vivo study reports a conjunctival redness score of 2 in 2 of the 3 animals (mean of individual scores at 24, 48 and 72 hours).

Justification for classification or non-classification

Based on the available data and according to the CLP regulations, the test item should not be classified as a skin irritant. However, the substance is observed to be category 2 eye irritant (H319) according to the criteria of the CLP Regulation) based on the fact that a conjunctival redness score of 2 was observed in 2 of the 3 animals (mean of individual scores at 24, 48 and 72 hours).