Registration Dossier

Administrative data

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

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1990
Report date:
1990

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
Trisodium bis[3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-2-hydroxy-5-nitrobenzenesulphonato(3-)]cobaltate(3-)
EC Number:
282-438-4
EC Name:
Trisodium bis[3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-2-hydroxy-5-nitrobenzenesulphonato(3-)]cobaltate(3-)
Cas Number:
84204-70-6
Molecular formula:
C32H20CoN10Na3O14S2
IUPAC Name:
trisodium bis[3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-2-hydroxy-5-nitrobenzenesulphonato(3-)]cobaltate(3-)
Test material form:
solid: particulate/powder
Details on test material:
Environmental fate/Ecotoxicological/Toxicological studies:
Storage: Room temperature, 20-24 °C, in the dark
Stability: pure, for 5 years; in solvent > 24 hours in water, DMSO, DMF, stable for at least 2 hours in aqueous medium.
Evidence of chemical instability at pH 6: none
Safety precaution: routine hygienic procedures were sufficient to assure personnel health and safety.
Toxicological studies:
Preparation: On the day of the experiment, the test article was suspended in Carboxymethylcellulose (1 %).
The vehicle was chosen to its nontoxicity for the animals. All animals received a single standard dose volume of 10 mL/kg body weight orally.

Test animals / tissue source

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.

Test system

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.

Results and discussion

In vivo

Resultsopen allclose all
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.

Any other information on results incl. tables

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.

Applicant's summary and conclusion

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.