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Administrative data

Description of key information

The undiluted test material produced moderate skin irritating effects, but the mean skin irritation scores (erythema and oedema) were below the threshold for classification.

The undiluted test material produced slight to well-defined signs of irritation to the rabbit eye, but the mean eye irritation scores (cornea opacity, iris, conjunctivae and chemosis) were below the threshold for classification.

According to the criteria described in the CLP regulation, the test item shall not be classified as skin or 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
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Principles of method if other than guideline:
The test item was applied once to the intact skin under semi-occlusive patches for 4 hours. After removal of the patches and wiping the treated areas the skin was examined for irritation. Further assessments were made 24, 48 and 72 hours after treatment.
GLP compliance:
not specified
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: 9-12 weeks
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL
- Concentration (if solution): undiluted

POSITIVE & NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 0.5 mL for solutions or 0.2 g for solids
- Concentration (if solution): undiluted
Duration of treatment / exposure:
4 hours
Observation period:
72 hours
Number of animals:
8
Details on study design:
TEST SITE
- Area of exposure: 3 x 3 cm
- % coverage: 100%
- Type of wrap if used: canvas body sleeve

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes
- Time after start of exposure: 4 hours

OBSERVATION TIME POINTS: 24, 48 and 72 hours

SCORING SYSTEM:
- Method of calculation: 8-point anchored ordinate scale ranging from 'a' (very slight) to 'h' (severe). At the end of the test the reaction grades are converted to numerical scores to calculate the total irritation score per site.
Irritation parameter:
erythema score
Basis:
mean
Time point:
24/48/72 h
Score:
1.83
Irritation parameter:
edema score
Basis:
mean
Time point:
24/48/72 h
Score:
2.04

Individual skin irritation scores

8-point anchored ordinate scale:

Rabbit N° 24 hours 48 hours 72 hours
erythema oedema erythema oedema erythema oedema
1 d e d d b c
2 c c c c a b
3 d c d d d c
4 d d d d d d
5 d d c c b c
6 c c d c b b
7 c e c d c d
8 d e d d c d

a very slight
b slight
c fairly distinct
d quite distinct
e becoming well developed
f well developed
g becoming severe
h severe

Numerical scores to calculate the total irritation score:

Rabbit N° 24 hours 48 hours 72 hours Average
erythema oedema erythema oedema erythema oedema erythema oedema
1 2 3 2 2 1 2 1,67 2,33
2 2 2 2 2 1 1 1,67 1,67
3 2 2 2 2 2 2 2,00 2,00
4 2 2 2 2 2 2 2,00 2,00
5 2 2 2 2 1 2 1,67 2,00
6 2 2 2 2 1 1 1,67 1,67
7 2 3 2 2 2 2 2,00 2,33
8 2 3 2 2 2 2 2,00 2,33
Average 1,83 2,04
Interpretation of results:
GHS criteria not met
Conclusions:
The substance is not skin irritant.
Executive summary:

In the current study the irritation effects of the test substance were investigated in rabbits. The study was not according to an OECD TG, however, it is similar to OECD 404.

The test item was applied undiluted to the clipped and intact skin under semi-occlusive patches for 4 hours. After removal of the patches and wiping the treated areas irritation reactions were examined immediately. Further assessments were made 24, 48 and 72 hours after treatment. Positive (Geranial and cyclamen aldehyde) and negative controls (diethyl phthalate) were tested in parallel.

The reaction grades for each treatment group were converted to corresponding numerical scores that were used to calculate the total irritation score per animal.

Under the conditions of this study, the undiluted test material produced a moderately irritating effect to rabbit skin, which was still visible after 72 hours treatment. The positive controls, geraniol and cyclamen aldehyde, produced reactions much more severe and the control diethyl phthalate showed no visible signs after 72 hours.

According to the classification criteria as stipulated in CLP, a classification as a skin irritant is required in case the mean erythema or oedema score over 24, 48 and 72 hours is at or above 2.3 in at least 2 out of 3 animals. The individual scores reported in this study indicates that all erythema scores are < 2.3. Out of 8 animals, 3 rabbits have an oedema score of 2.33. All other animals (5 out of 8) have an oedema score < 2.3. From these observations it can be concluded that the test item does not require classification as a skin irritant in accordance with the 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:
15.05 - 23.05.2000
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
other: SPF albino rabbits of the stock Chbb:HM(SPF) - Littlerussian
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Weight at study initiation: 2.8 - 3.0 kg b.w.
- Housing: individually in PPO cages (floor area: 2576 sq.cm) with perforated floor.
- Diet: pelleted complete rabbit diet "Altromin 2123", ad libitum
- Water: ad libitum, domestic quality drinking water acidified with hydrochloric acid to pH 2.5 to prevent microbial growth
- Acclimation period: 1 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20°C ± 3°C
- Humidity (%): 55% + 15%
- Air changes (per hr): 10 times/hour
- Photoperiod (hrs dark / hrs light): Light was on from 6 a.m. to 6 p.m..
Vehicle:
not specified
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL of undiluted test item
Duration of treatment / exposure:
24 hours
Observation period (in vivo):
7 days
Number of animals or in vitro replicates:
4
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): 20 ml 0.9% sodium chloride solution
- Time after start of exposure: 24 hours

SCORING SYSTEM: See 'Any other information on materials and methods'

TOOL USED TO ASSESS SCORE: fluorescein
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Irritation parameter:
iris score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
24/48/72 h
Score:
1
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
mean
Time point:
24/48/72 h
Score:
0.9
Reversibility:
fully reversible within: 7 days
Irritant / corrosive response data:
Animals 1 and 4 showed markedly deepened folds, congestion, swelling, moderate circumcorneal injection, reaction to light, some injection of conjunctival vessels and discharge different from normal 1 hour after application. Animal 2 showed a diffuse, crimson red conjunctiva with individual vessels not easily discernible as well as obvious swelling with partial eversion of lids. Animal 3 showed some conjunctival vessels definitely injected, swelling with partial eversion of lids and discharge different from normal.
24 hours after application animals 1 and 2 showed some conjunctival vessels definitely injected and swelling above normal. Animals 3 and 4 showed diffuse, crimson red conjunctiva with individual vessels not easily discernible as well as obvious swelling with partial eversion of lids.
48 hours after application all animal showed some injected conjunctival vessels and animals 2, 3 and 4 showed additionally swelling above normal.
72 hours after application animals 3 and 4 showed some injected conjunctival vessels and swelling above normal. Animals 1 and 2 were free of any signs of eye irritation.
7 days after application all animals were free of any signs of eye irritation.

Individual irritation scores

Animal n°

Cornea opacity

Iris lesion

Redness of conjunctiva

Oedema of conjunctiva

1

0.00

0.00

0.67

0.33

2

0.00

0.00

0.67

0.67

3

0.00

0.00

1.33

1.33

4

0.00

0.00

1.33

1.33

Mean values

0.00

0.00

1.0

0.9

Interpretation of results:
GHS criteria not met
Conclusions:
The test item shall not be classified as eye irritating.
Executive summary:

The objective of this study was to assess the potential eye irritant effect of the test substance according to the method in the OECD Guideline 405 and EEC Guideline B.5.

The study was initiated with one rabbit. Since no marked eye irritation was seen in this animal three other rabbits were included in the study with time difference.

The female albino rabbits were exposed to 0.1 ml of the test article in the left eye while the right eye served as control. The eyes were examined and the changes were graded according to a numerical scale 1, 24, 48 and 72 hours as well as 7 days after dosing.

Slight to well-defined signs of irritation were observed in the treated eyes, however, all effects were fully reversible within 7 days.

The following mean values for all animals, based an the results from the 24, 48 and 72 hour readings, were calculated:

- cornea opacity score: 0.0

- iris score: 0.0

- conjunctivae score: 1.0

- chemosis score: 0.9

According to the CLP regulation, the test item shall not be classified as eye irritating.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Additional information

Skin irritation

For the endpoint skin irritation 1 in vivo study is available (1980). In this study the skin irritation effects were investigated in rabbits and the study was similar to OECD 404. The test item was applied undiluted to clipped and intact skin under semi-occlusive patches for 4 hours. After removal and washing skin reactions were assessed immediately, 24, 48 and 72 hours after treatment. Positive and negative controls were tested in parallel.

The undiluted test material produced moderate irritating effects that were still visible after 72 hours of treatment. However, the mean erythema score of 8 animals was 1.83 and the mean oedema score of 8 animals was 2.04, indicating that the substance is not to be considered as skin irritant according to the CLP regulation.

Eye irritation

For eye irritation there is 1 in vivo study available (2000). In this study eye irritant effects of the test substance were assessed in rabbits and the test was according to OECD 405. The study was initiated with one rabbit and no marked eye irritation was seen. 3 rabbits were included in the study and exposed to 0.1 ml of the undiluted test article in the left eye and the eyes were examined 1, 24, 48 and 72 hours as well as 7 days after dosing.

Slight to well-defined signs of irritation were observed in the treated eyes, however, all effects were fully reversible within 7 days. The mean cornea opacity score was 0.0, the mean iris score was 0.0, the mean conjunctivae score was 1.0 and the mean chemosis score was 0.9. According to the CLP regulation, the test item shall not be classified as eye irritating.

Taking together, the substance is not irritant to the skin or the eye.

Justification for classification or non-classification

Skin irritation

The observed effects of skin irritation are slight, with a mean erythema score of 8 animals after 24, 48 and 72 hours of 1.83 and a mean oedema score of 8 animals after 24, 48 and 72 hours of 2.04.

According to the EU Regulation No. 1272/2008 on the Classification, Labelling and Packaging of Substances and Mixtures (CLP) a substance is considered skin irritant when the mean value is >= 2.3 and =< 4 for erythema/eschar or oedema in at least 2 out of 3 animals from gradings at 24, 48 and 72 hours after the removal of the patch is observed. Based on the mean scores for the individual animals reported in the key study, classification of the test substance is not required according to the criteria set out in the CLP regulation.

Eye irritation

The observed mean of 4 animals for cornea opacity score was 0.0, iris score was 0.0, conjunctivae score was 0.0 and chemosis score was 0.9.

According to the EU Regulation No. 1272/2008 on the Classification, Labelling and Packaging of Substances and Mixtures (CLP) a substance is considered eye irritant when at least in one animal effects on the cornea, iris or conjunctiva that are not expected to reverse or have not fully reversed within an observation period of normally 21 days; and/ or at least in 2 of 3 tested animals, a positive response of corneal opacity >= 3 and/or iritis > 1,5, calculated as the mean scores following grading at 24, 48 and 72 hours after installation of the test material.

Therefore, classification of the test substance is not required according to the criteria set out in the CLP regulation.