Trisodium bis[4-[4,5-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-3-methyl-5-oxo-1H-pyrazol-1-yl]benzene-1-sulphonato(3-)]cobaltate(3-)

Endpoint summary

Description of key information

Skin: not irritant
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:

No guideline followed; 6 rabbits tested for skin irritation on intact and wound skin.

GLP compliance:

no

Species:

rabbit

Preparation of test site:

other: intact and wound

Amount / concentration applied:

0.5 g

Duration of treatment / exposure:

72 h

Observation period:

24 and 72 h

Number of animals:

6

Irritation parameter:

erythema score

Remarks:

intact and wound skin

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Remarks:

intact and wound skin

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Interpretation of results:

other: not classified within the CLP Regulation (EC 1272/2008)

Conclusions:

Not irritant to the skin.

Executive summary:

Skin irritation test was performed on 6 rabbits on both intact and wound skin. 0.5 g of test substance was applied. Observations for effects in terms of erythema and oedema were done at 24 and 72 h after start of application.

Irritation score was 0 for all rabbits.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Principles of method if other than guideline:

No guideline followed; the substance is applied to the eye of one rabbit and observations are done after 24, 48, 72 h and 7 d from start of exposure. Effects on both washed and unwashed eye are recorded.

Species:

rabbit

Observation period (in vivo):

24, 48, 72 h and 7 d for both washed and unwashed eye.

Number of animals or in vitro replicates:

1 rabbit

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

animal #1

Remarks:

unwashed eye

Time point:

24/48/72 h

Score:

0.67

Max. score:

1

Reversibility:

fully reversible within: 72 h

Irritation parameter:

conjunctivae score

Basis:

animal #1

Remarks:

washed eye

Time point:

24/48/72 h

Score:

0.67

Max. score:

1

Reversibility:

not fully reversible within: 7 d

Other effects:

reddening of eye; immediately violent pain.

Interpretation of results:

study cannot be used for classification

Conclusions:

Evidence of reddening in treated eye not completely reversible within 7 days.

Executive summary:

The eye irritation potential of test substance was assessed in eyes of one rabbit; one eye was washed, the other was unwashed.

As the rabbit immediately showed a pain reaction, only one rabbit was used.

Effects on iris, cornea and conjunctivae were assessed after 24, 48, 72 hours and 7 days. Only conjunctivae redness was noted and was scored 1. In the washed eye, such effects was not reversible within the 7 days observation period.

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

From April 13th to May 1st, 1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Details on the read-across are available in section 13.

Reason / purpose for cross-reference:

read-across source

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)

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 male, 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 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.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 g of test substance

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 examined 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 Safety 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:

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:

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:

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:

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:

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:

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:

24/48/72 h

Score:

2

Reversibility:

fully reversible within: 14 days

Remarks on result:

other: not rinsed

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2

Reversibility:

fully reversible within: 14 days

Remarks on result:

other: not rinsed

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2

Reversibility:

fully reversible within: 14 days

Remarks on result:

other: not rinsed

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

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:

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:

24/48/72 h

Score:

1

Reversibility:

fully reversible within: 14 days

Remarks on result:

other: not rinsed

Irritant / corrosive response data:

No vascularization was observed.
No acute clinical symptoms were observed in the animals during the test period and no mortality occurred.
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.

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).

Interpretation of results:

other: classified as irritant within the CLP Regulation (EC 1272/2008)

Conclusions:

Irritant for rabbit eye.

Executive summary:

The substance is tested for eye irritation according to OECD guideline 405.

Observation for effects was done 1, 24, 48 and 72 hours after start of exposure, up to 7 and 14 days. It was terminated after 14 days, because a clear tendency of a healing process was observed.

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

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

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

Mean value of the scores for each type of lesion, calculated for each animal separately, is the following:

1 male

Cornea opacity: 2

Iris: 0.67

Conjunctivae redness: 2

Conjuntivae chemosis: 2

1 male

Cornea opacity: 1.67

Iris: 0.67

Conjunctivae redness: 2

Conjuntivae chemosis: 2

1 female

Cornea opacity: 1

Iris: 0

Conjunctivae redness: 2

Conjuntivae chemosis: 1

Based on test results, the substance has to be considered as irritant to eyes.

As treated eyes were not rinsed, effects represent a worst-case situation.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Acid Brown 303 was found to be completely not irritant to rabbit skin. The substance was applied on tboth intcat and wound skin of 6 rabbits and no effects in terms of erythema and oedema were seen.

In addition, data on a structural analogue, i.e. Similar Substance 01, was used to support findings on Acid Brown 303. Details on the read-across are available in section 13.

Skin irritation was tested following OECD guideline 404 and no signs of irritation were recorded.

Acid Brown 303 was tested for eye irritation on one single rabbits. Only one single rabbit was used; pain reactions were reported soon after application. No effect on cornea, iris and conjuctivae chemosis was seen; effect as conjunctivae redness was seen. Such effect was not completely reversible up to the last observation on day 7.

Due to the lack of details test method and results, the reliability of the study could not be ascertained and the study could not be used to the purposes of classification.

Accordingly, available data on a structural analogue, i.e Similar Substance 01, was used. Deatils on the read-across are available in section 13.

Similar Substance 01 was tested following OECD guideline 405. Effects were recorded at 24, 48 and 72 h post exposure and up to day 7 and 14. Treated eyes were not rinsed.

Mean score (24/48/72 h) in all rabbits was >= 1 for corneal opacity and equal to 2 for conjunctivae redness; effects on conjunctivae chemosis were scored 2 in 2/3 rabbits; effects on iris were scored < 1 in all rabbits. All effects were completely reversible by the end of the observation period.

Justification for classification or non-classification

According to the CLP Regulation (EC 1272/2008), classification as skin irritant (category 2) applies if the substance produces:

- 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

- 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

- 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.

In available studies, no signs of irritation were seen, thus Acid Brown 303 is not classified as a skin irritant.

 

According to the CLP Regulation (EC 1272/2008), classification as eye irritant (category 2) applies if the substance produces in at least 2/3 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.

Based on experimental findings in the key study, scores for effects observed are above the reference thresholds in at least 2/3 animals. Such effects are fully reversible within 14 days.

As treated eyes were not rinsed upon exposure to test substance, test conditions represent a worst-case possibly influencing the study outcome.

By following a worst-case approach, Acid Brown 303 is classified as eye irritant, category 2 (H319) within the CLP regulation (EC 1272/2008).