Registration Dossier

Administrative data

Description of key information

Skin: not irritating, not corrosive
Eye: cause serious eye irritation

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation / corrosion, other
Remarks:
points 8.1 and 8.2 of Annex VIII of REACH have been amended. Nevertheless, adequate information from existing in vivo skin irritation or eye irritation studies can still be used to fulfil the information requirement at any tonnage level.
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From the 13th to the 20th of April, 1990
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Study conducted according to internationally accepted guideline
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
Source: BRL, Biological Research Laboratories Ltd. Wolferstrasse 4, CH-4414 Fullinsdorf
Number of animals: 1 male, 2 females
Age at start of treatment: male, 15 weeks, females 16 weeks
Body weight at start of or treatment: male, 3.0 kg, females: 3.2 kg
Identification: by unique cage number and corresponding ear tags.
Acclimatization: four days under test conditions after veterinary examination.
Allocation: Animal Numbers, Male no, 1 Female nos. 2, 3

HUSBANDRY
Room No,: 138
Standard Laboratory Conditions.
Air-onditioned with 10-15 air changes per hour and hourly monitored environment with temperature 20±3 °C, relative humidity 40-70%, 12 hours
artificial fluorescent light/12 hours dark, music/light period.
Accommodation: individually in stainless steel cages equipped with an automatic cleaning and drinking system.
Diet: pelleted standard Kliba 341, Batch 52/90 rabbit maintenance diet (“Kliba”, Klingentalmuehle AG, CH-4303 Kaiseraugst), ad libitum,
Analysis for contaminants performed.
Water: community tap water from Itingen, ad libitum
Analysis for contaminants performed.
Type of coverage:
semiocclusive
Preparation of test site:
shaved
Controls:
no
Amount / concentration applied:
0.5 g of the test article
Duration of treatment / exposure:
4 hours
Observation period:
Viability, mortality: daily
Body weight: Start of acclimatization, day 1 of test (application day) and at termination of observation.
The skin reaction was assessed at 1, 24, 48 and 72 hours intervals after the removal of the dressing, gauze patch and test article.
Number of animals:
1 male
2 females
Details on study design:
Approximately 24 hours before treatment, the dorsal fur was shaved with an electric clipper, exposing an area of approximately 100 square centimeters (10 cm x 10 cm). The skin of the animals was observed 1 day prior to test article administration.
On test day 1, 0.5 g of the test article was applied to approx. 6 cm^2 of the intact skin of the shaved area. It was covered with a 3.0 cm x 3.0 cm patch of surgical gauze. The gauze was covered with a semi—occlusive dressing. The dressing was wrapped around the abdomen and anchored with an elastic bandage.
Four hours after the application, the dressing was removed and the skin was flushed with lukewarm tap water. The skin reaction was assessed according to the following numerical scoring system.
The observation was terminated 72 hours after application of the test article.
All rabbits were killed by an intravenous injection of T61 (Hoechst AG) into the ear vein and discarded.
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
edema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
edema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
edema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Reversibility:
fully reversible within: 72 hours
Irritant / corrosive response data:
The substance showed a primary irritation score of 0.00 when applied to intact rabbit skin. Local signs (mean 1 hour to 72 hours/day) consisted or grade 0.00 erythema and grade 0.00 edema.
No destructions or irreversible alterations or the treated skin were observed. No corrosive effect was evident on the skin.
In the area of application yellowish staining of the treated skin by pigment or coloring of the test article was observed in all animals between 1 to 72 hours
(termination or the study).
Other effects:
The body weight gain of all rabbits was similar.
No acute clinical symptoms were observed in the animals during the test period, and no mortality occurred.
No necropsy was performed. All animals were killed and discarded.

Data are summarized in tabular form, showing for each individual animal the irritation scores for erythema and edema at 60 minutes, 24, 48 and 72 hours after patch removal, any lesions, a description or the degree and nature of irritation, corrosion or reversibility, and any other toxic effects observed. The dermal irritation scores were evaluated in conjunction with the nature and reversibility or otherwise of the responses observed. The individual scores do not represent an absolute standard for the irritant properties of a material, and they are viewed as reference values. The use of a semi-occlusive dressing is a severe test and the results are not directly comparable to human exposure conditions.

Mean values and standard deviations were calculated whenever possible.

Interpretation of results:
not classified
Remarks:
Classification criteria according to the CLP Regulation 1272/2008 and its amendments
Conclusions:
Non irritant/corrosive for the skin
Executive summary:

Materials and methods

The irritation/corrosion property was tested on the substance according to the OECD Guideline 404 (Acute Dermal Irritation/Corrosion) and the EU Method B.4 (Acute Toxicity: Dermal Irritation/Corrosion), in GLP.

Under the conditions of the experiment, the tested substance was found to cause a primary irritation score of 0.00 when applied to intact rabbit skin. No corrosive effect had occurred on the skin at any measuring interval.

Local signs (mean 1 hour to 72 hours) consisted of grade 0.00 erythema and grade 0.00 edema.

In the area of application, yellowish staining of the treated skin by pigment or coloring of the test article was observed in all animals until termination.

Observations

According to EEC Council Directive, 83/467/EEC, the mean value of the scores for each type of lesion, calculated for each animal separately, is the following:

Mean 24-72 hours 0.00 (Erythema, Edema), all the animals (1 male, 2 females).

Conclusions

Due to the results described above, the substance is considered non irritant to the skin of rabbits.

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

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation, other
Remarks:
points 8.1 and 8.2 of Annex VIII of REACH have been amended. Nevertheless, adequate information from existing in vivo skin irritation or eye irritation studies can still be used to fulfil the information requirement at any tonnage level.
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From the 13th of April to the 1th of May, 1990
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Study conducted according to internationally accepted guideline
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
Source: BRL, Biological Research Laboratories Ltd. Wolferstrasse 4, CH-4414 Fullinsdorf
Number of animals: 2 malel, 1 female
Age at start of treatment: male, 15 weeks, females 16 weeks
Body weight at start of or treatment: male 2.7-2.9 kg, females: 2.3 kg
Identification: by unique cage number and corresponding ear tags.
Acclimatization: four days under test conditions after veterinary examination.
Allocation: Animal Numbers, Male no 96-97 Female nos. 98

HUSBANDRY
Room No: 138
Standard Laboratory Conditions:
Air-onditioned with 10-15 air changes per hour and hourly monitored environment with temperature 20±3 °C, relative humidity 40-70%, 12 hoursartificial fluorescent light/12 hours dark, music/light period.
Accommodation: individually in stainless steel cages equipped with an automatic cleaning and drinking system.
Diet: pelleted standard Kliba 341, Batch 52/90 rabbit maintenance diet (“Kliba”, Klingentalmuehle AG, CH-4303 Kaiseraugst), ad libitum,
Analysis for contaminants performed.
Water: community tap water from Itingen, ad libitum
Analysis for contaminants performed.
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
0.1 g of the test article
Observation period (in vivo):
Viability, mortality: daily
Body weight: Start of acclimatization, day 1 of test (application day) and at termination of observation.
The eye reaction were wxamined at 1, 24, 48 and 72 hours, 7 and 14 days after administration.
Number of animals or in vitro replicates:
2 males, 1 female
Details on study design:
The eyes of the animals were observed one day prior to test article administration.
On test day 1, 0.1 g or the test article was placed in the conjunctival sac or the left eye of each animal after gently pulling the lower lid away from the
eyeball. The lids were then gently held together ror about one second to prevent loss or the test article. The right eye remained untreated and served as the reference control.
In cases of equivocal results when comparing the treated and untreated eyes, the illustrated guide rrom the Consumer Product Sarety Commission Washington, D.C. 20207 would be used for additional control purposes.
Eye examinations were made with a slit—lamp 30 SL/M (C. Zeiss AG, Zuerich/Switzerland) and a Varta Cliptrix diagnostic—lamp (A. Riegger, Basel/
Switzerland).
The observation was terminated 14 days after administration of the test article.
No necropsy was performed in the animals killed at termination of observation. All rabbits were killed by an intravenous injection of T61 (Hoechst/AG) into the ear vein and discarded.
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
other: 24, 48, 72 h
Score:
2
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: not rinsed
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
other: 24,48, 72 h
Score:
1.6
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: not rinsed
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
other: 24, 48, 72 h
Score:
1
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: not rinsed
Irritation parameter:
iris score
Basis:
animal #1
Time point:
other: 24, 48, 72 h
Score:
0.67
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: not rinsed
Irritation parameter:
iris score
Basis:
animal #2
Time point:
other: 24, 48,72 h
Score:
0.67
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: not rinsed
Irritation parameter:
iris score
Basis:
animal #3
Time point:
other: 24,48, 72 h
Score:
0
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: not rinsed
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
other: 24,48, 72 h
Score:
2
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: redness, not rinsed
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
other: 24, 48, 72 h
Score:
2
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: redness, not rinsed
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
other: 24, 48, 72 h
Score:
2
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: redness, not rinsed
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: 24, 48, 72 h
Score:
1.33
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: not rinsed
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
other: 24, 48, 72 h
Score:
1
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: not rinsed
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
other: 24, 48, 72 h
Score:
1
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: not rinsed
Irritant / corrosive response data:
Primary Irritation score: 4.58 (not rinsed). No vascularization was observed.
No acute clinical symptoms were observed in the animals during the test period, and no mortality occurred.
The substance showed a primary irritation score of 4.58 when applied to the conjunctival sac of the rabbit eye.
In the area or application a yellow staining of the cornea and conjunctivae by pigment or coloring of the test article was observed.
Opacity was observed in two animals between 1 hour and 7 days and in one animal between 1 to 72 hours after test article administration.
Other effects:
The body weight gain of all rabbits was similar.
No necropsy was performed. All animals were killed and discarded.

Data will be summarized in tabular form, showing for each individual animal the irritation scores at the designated observation time, a description of the degree and nature of irritation, the presence or serious lesions and non-ocular effects.

To calculate the irritation effect all scores of each animal at each of the following reading times (24, 48, 72 hours) were used in calculating the respective mean values (Council Directive 83/467/EEC, July 1983 and 67/548/EEC, May 1987, Brussels, Belgium).

The ocular irritation scores were evaluated in conjunction with the nature and reversibility or otherwise or the responses observed. The individual scores do not represent an absolute standard for the irritant properties of a material, and they are viewed as rererence values and are only meaningrul when supported by a full description and evaluation of these observations.

Mean values and standard deviations were calculated whenever possible.

Interpretation of results:
other: Eye irrit. cat 2: Causes serious eye irritation
Remarks:
Classification criteria according to the CLP Regulation 1272/2008 and its amendments
Conclusions:
Irritating for the rabbit eye
Executive summary:

Materials and methods

The eye irritation/corrosion property was tested on the substance according to the OECD Guideline 405 (Acute Eye Irritation / Corrosion) and the EU Method B.5 (Acute Toxicity: Eye Irritation/Corrosion), in GLP.

Under the conditions of this experiment, the substance was found to cause a primary irritation score or f 4.58, when applied to the conjunctival sac of the rabbit eye.

The observation was terminated after 14 days, because a clear tendency of a healing process was observed.

A yellow staining of the cornea and conjunctivae of’ the treated eyes by pigment or coloring of the test article was observed.

If necessary prior to the first reading of the reactions, the eye was cleaned with disposable tissues to remove staining produced by the test article, so that the reactions (erythema) were clearly visible at that time.

Opacity of cornea was observed in two animals between 1 hour and 7 days and in one animal between 1 to 72 hours.

According to EEC Council Directive, 83/467/EEC, July 1983 and 67/548/EEC, May 1987, Brussels, Belgium, the mean value of the scores rot each type of lesion, calculated for each animal separately, is the following:

rabbit n° h opacity Iris redness chemosis
96 24 2 1 2 3
48 2 1 2 1
72 2 0 2 0
mean 2 0.66 2 1.33
97 24 2 1 2 2
48 2 1 2 1
72 1 0 2 0
mean 1.66 0.66 2 1
98 24 1 0 2 2
48 1 0 2 1
72 1 0 2 0
mean 1 0 2 1

Due to the results described above, the test article had to be classified as irritating to the eyes.

This classification based on the fact that generally diffuse corneal opacity was observed in two animals between 1 hour and 7 days and in one animal between 1 to 72 hours of observation. No vascularization was observed. It should be noted that no rinsing of the eye has been applied.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation/corrosion

The irritation/corrosion property was tested on the substance according to the OECD404 and EU B.4, in GLP.

Under the conditions of the experiment, the tested substance was found to not cause corrosion or irritating effects.

Due to the results described above, the substance is considered non irritant to the skin of rabbits.

Eye irritation/corrosion

The eye irritation/corrosion property was tested on the substance according to the OECD 405 and EU B.5, in GLP.

Under the conditions of this experiment, the substance was found to cause a primary irritation score or f 4.58, when applied to the conjunctival sac of the rabbit eye. An irritant effect was observed, but fully reversible after 14 days.

According to EEC Council Directive, 83/467/EEC, July 1983 and 67/548/EEC, May 1987, Brussels, Belgium, the mean value of the scores was calculated, indicating that the test article had to be classified as irritating to the eyes.:

rabbit n° h opacity Iris redness chemosis
96 24 2 1 2 3
48 2 1 2 1
72 2 0 2 0
mean 2 0.66 2 1.33
97 24 2 1 2 2
48 2 1 2 1
72 1 0 2 0
mean 1.66 0.66 2 1
98 24 1 0 2 2
48 1 0 2 1
72 1 0 2 0
mean 1 0 2 1

Justification for classification or non-classification

Skin irritation/corrosion

Substances that have the potential to induce reversible skin irritation are classified in Category 2 (irritating to skin).

According to the CLP Regulation 1272/2008/EC, the substance can be classify as a single irritant category (Category 2) according to the criteria, section 3.2, table 3.2.2.

If, when applied to the skin of an animal, the substance produces:

(1) Mean value of ≥ 2,3 - ≤ 4,0 for erythema/eschar or for oedema in at least 2 of 3 tested animals from gradings at 24, 48 and 72 hours after patch removal or, if reactions are delayed, from grades on 3 consecutive days after the onset of skin reactions; or

(2) Inflammation that persists to the end of the observation period normally 14 days in at least 2 animals, particularly taking into account alopecia (limited area), hyperkeratosis, hyperplasia, and scaling; or

(3) In some cases where there is pronounced variability of response among animals, with very definite positive effects related to chemical exposure in a single animal but less than the criteria above. Reversibility of skin lesions is the other decisive factor in evaluating responses in the animal test.

Under the conditions of the experiment, the tested substance was found to cause a primary irritation score of 0.00 when applied to intact rabbit skin. No corrosive effect had occurred on the skin at any measuring interval.

Local signs (mean between 1 hour to 72 hours) consisted of grade 0.00 erythema and grade 0.00 edema.

According to the paragraph 3.2. of the CLP Regulation n. 1272/2008, the substance is not classified as a skin irritant/skin corrosive.

 

Eye irritation/corrosion

Substances that have the potential to induce reversible eye irritation are classified in Category 2 (irritating to eyes).

According to the CLP Regulation 1272/2008/EC, using the results of animal testing the substance can be classify as a single irritant category (Category 2) according to the criteria, section 3.3, table 3.3.2

If, when applied to the eye of an animal, the substance produces:

– at least in 2 of 3 tested animals, a positive response of:

corneal opacity ≥ 1 and/or iritis ≥ 1, and/or

conjunctival redness ≥ 2 and/or

conjunctival oedema (chemosis) ≥ 2

calculated as the mean scores following grading at 24, 48 and 72 hours after installation of the test material, and which fully reverses within an observation period of 21 days.

The substance produced a positive response ≥ 1 on corneal opacity,

= 2 on conjunctival redness

= 1.11 conjunctival oedema (chemosis)

in two or three animals, calculated as the mean of scores at 24, 48, 72 hours.

The damage was fully reversible after 14 days, therefore the substance should be classified, according to the CLP Regulation 1272/2008/EC, as Irritant, Category 2, H319.