Tetrasodium 5-[4-chloro-6-(N-ethyl-anilino)-1,3,5-triazin-2-ylamino]-4-hydroxy-3-(1,5-disulfonatonaphthalen-2-ylazo)-naphthalene-2,7-disulfonate

Endpoint summary

Description of key information

Reactive Red 245 is not irritating to the skin, but has irreversible effects on the eye.

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:

25 Sep 1990 to 02 Nov 1990

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

GLP compliance:

yes

Remarks:

OECD Principles of Good Laboratory Practice, Paris, France

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: SR 6947/14.
- Expiration date of the lot/batch: July, 1995.

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: in the original container, protected from light, at room temperature (approx. 20°C).
- Stability under test conditions: stable for at least 48 hours.

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd., Wölferstrasse 4, CH-4414 Füllinsdorf
- Age at start of treatment: Male: 15 weeks, Females: 16 weeks
- Weight at start of treatment: Male: 2.9 kg, Females: 2.9 - 3.1 kg
- Housing: Individually in stainless steel cages equipped with an automatic cleaning and drinking system.
- Diet: Pelleted standard Kliba 341, batch 55/90 rabbit maintenance diet ("Kliba", Klingentalmuehle AG, CH-4303 Kaiseraugst), ad libitum
- Water: Community tap water from Itingen ad libitum
- Acclimation period: Four days under test conditions after veterinary examination

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 40-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Remarks:

Test material was applied moistened in distilled water

Controls:

no

Amount / concentration applied:

0.5 g

Duration of treatment / exposure:

4 h

Observation period:

up to 72 h

Number of animals:

3 (1 male and 2 females)

Details on study design:

Approximately 24 h before treatment, the dorsal fur was clipped with an electric clipper, exposing an area of approximately 100 square centimeters (10 cm x 10 cm). The skin of the animals was examined once, 1 day prior to test article administration. On test day 1, 0.5 g of the test article was applied to approximately 6cm² of the intact skin of the clipped 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 at 1, 24, 48 and 72 h intervals after the removal of the dressing, gauze patch and test article.

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Irritation: FAT 40406/A showed a primary irritation score (24, 48, 72 h) of 0.00 when applied to intact rabbit skin. Local signs (mean 1 h to 72 h) consisted of grade 0.00 erythema and grade 0.00 edema.
Corrosion: No destructions or irreversible alterations of the treated skin were observed. No corrosive effect was evident on the skin.

Other effects:

Coloration: In the area of application a red staining of the treated skin by pigment or coloring of the test article was observed during the observation period.
Body weights: The body weight gain of all rabbits was similar.
Toxic symptoms/mortality: No acute clinical symptoms were observed in the animals during the test period, and no mortality occurred.

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.

 

Necropsy:

The observation was terminated 72 hours after application of the test article. No necropsy was performed in the animals euthanized at termination of

observation. All rabbits were euthanized by an intravenous injection of T61 (Hoechst AG) into the ear vein and discarded.

Interpretation of results:

GHS criteria not met

Conclusions:

FAT 40406/A was found to be not irritating or corrosive to skin.

Executive summary:

In a GLP-compliant skin irritation study, performed according to OECD guideline 404, the test substance (moistened, 0.5 g) was applied, semi-occlusive, to the skin of three New Zealand White rabbits for 4 h. After 4 h, the treated skin was flushed with water and observation were made for an additional 72 h. The test substance showed a primary irritation score (24, 48, 72 h) of 0.00 and local signs (1 hour to 72 hours) consisted of grade 0.00 erythema and grade 0.00 edema. No corrosive effect was evident on the skin. Under the conditions of this experiment, FAT 40406/A was found to be not irritating or corrosive to skin.

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:

5 Oct 1990 to 2 Nov 1990

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

GLP compliance:

yes

Specific details on test material used for the study:

Name: FAT 40406/A
Purity: 89.3 %

SOURCE OF TEST MATERIAL
- Source and lot/batch No. of test material: SR 6947/14
- Expiration date of the lot/batch: July 1995

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: in the original container, protected from light, at room temperature (approx. 20 °C)
- Stability under test conditions: stable

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd., Wölferstrasse 4, CH-4414 Füllinsdorf.
- Age at start of treatment: Males: 15 weeks, Female: 16 weeks.
- Weight at start of treatment: Males: 3-3.1 kg, Female: 2.8 kg.
- Housing: Individually in stainless steel cages equipped with an automatic cleaning and drinking system (Dipl. Ing. W. Ehret GmbH, Versuchstiertechnik, D-7830 Emmendingen).
- Diet: Pelleted standard Kliba 341, batch 56/90 rabbit maintenance diet ("Kliba", Klingentalmuehle AG, CH-4303 Kaiseraugst), ad libitum.
- Water: Community tap water from Itingen ad libitum.
- Acclimation period: Four days under test conditions after veterinary examination.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 40-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

0.1 g

Duration of treatment / exposure:

72 hours

Observation period (in vivo):

1, 24, 48, 72 h

Number of animals or in vitro replicates:

3 (2 males, 1 female)

Details on study design:

Both eyes of the animals were observed once, one day prior to test article administration. The test article (0.1 g) was placed in the conjunctival sac of the left eye of each animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test article. The right eye remained untreated and served as the reference control. The eyes of each animal were examined 1, 24, 48 and 72 h after administration. The irritation was assessed and the corrosive properties of the test article and the staining of the treated eye were described and recorded.

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

3

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0.22

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritant / corrosive response data:

Irritation: FAT 40406/A showed a primary irritation score of: 0.42 when applied to the conjunctival sac of the rabbit eye.
Corrosion: No corrosion of the cornea was observed at any of the measuring intervals.

Other effects:

Toxic symptoms/mortality: No acute clinical symptoms were observed in the animals during the test period, and no mortality occurred.

Coloration: A red staining of the conjunctivae, nictitating membrane, sclera and eyelashes of the treated eyes by pigment or coloring of the test article was observed in all animals from 1 hour until termination of observation. In addition a red staining of the cornea was observed from 24 h to 72 h and a red staining of the iris was observed at different hours according to the animals.

Body weights: The body weight gain of all rabbits was similar.

Interpretation of results:

GHS criteria not met

Conclusions:

FAT 40406/A has an irreversible effect on the eye.

Executive summary:

In a GLP-compliant eye irritation study, performed according to OECD guideline 405, the test substance (0.1 g) was placed in the conjunctival sac of the left eye of three New Zealand White rabbits. The eyes were examined for 72 h. The test substance showed a mean chemosis score of 1 and 0.67 after 1 and 24 h, respectively. The chemosis score was 0.00 at the other time points as were the cornea, iris, and conjunctivae scores during the whole observation period. The primary irritation score was 0.42. No corrosion of the cornea was observed at any of the measuring intervals. A red staining of the conjunctivae, nictitating membrane, sclera and eyelashes of the treated eyes by pigment or coloring of the test article was observed in all animals from 1 h until termination of observation. Therefore, it was concluded that the test substance had irreversible effects in the eye.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation/corrosion:

In a GLP-compliant skin irritation study, performed according to OECD guideline 404, the test substance (moistened, 0.5 g) was applied, semi-occlusive, to the skin of three New Zealand White rabbits for 4 h. After 4 h, the treated skin was flushed with water and observation were made for an additional 72 h. The test substance showed a primary irritation score (24, 48, 72 h) of 0.00 and local signs (1 h to 72 h) consisted of grade 0.00 erythema and grade 0.00 edema. No corrosive effect was evident on the skin. Under the conditions of this experiment, thetest substance FAT 40406/A was found to be not irritating or corrosive to skin.

 

Eye irritation/corrosion:

In a GLP-compliant eye irritation study, performed according to OECD guideline 405, the test substance (0.1 g) was placed in the conjunctival sac of the left eye of three New Zealand White rabbits (RCC 1990). The eyes were examined for 72 hours. The test substance showed a mean chemosis score of 1 and 0.67 after 1 and 24 hours, respectively. The chemosis score was 0.00 at the other time points as were the cornea, iris, and conjunctivae scores during the whole observation period. The primary irritation score was 0.42. No corrosion of the cornea was observed at any of the measuring intervals. A red staining of the conjunctivae, nictitating membrane, sclera and eyelashes of the treated eyes by pigment or coloring of the test article was observed in all animals from 1 hour until termination of observation.Therefore, it was concluded that the test substance had irreversible effects in the eye.

Justification for classification or non-classification

Based on the findings in the skin irritation study Reactive Red 245 does not need to be classified according to the Directive 67/548/EEC and according to the EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008 to be irritating to the skin. However, based on the findings in the eye irritation study, the substance need to be classified for Eye damage 1: H318 according to the EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.