Tetrasodium 3,3'-[[6-[(2-hydroxyethyl)amino]-1,3,5-triazine-2,4-diyl]bis[imino(2-methyl-4,1-phenylene)azo]]bisnaphthalene-1,5-disulphonate

Endpoint summary

Description of key information

In the available studies for skin irritation/corrosion and eye irritation bayscropt Gelb BR was not irritating/corrosive.

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:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Principles of method if other than guideline:

500 mg of the pulverized test substance were moistened with water (to ensure good contact with the skin) and subsequently applied to a hypoallergenic Hansamed® - patch. A further patch was moistened with water. The patches prepared in this way were placed on the opposite dorso-lateral areas of the trunk of each animal and were loosely held in place with a semiocclusive dressing for the duration of the exposure period. The treated skin area was approx. 6 cm² in size. After an exposure period of four hours, the dressing and patches were removed. The exposed skin areas were carefully washed with water and scored according to Draize.

GLP compliance:

yes

Specific details on test material used for the study:

State of Aggregation: solid
External Appearance: yellow-brown powder
contents: 100%
pH: 6-7 (40 g/L H2O)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Species and species justification
The study was conducted on rabbits an animal species recommended in the guidelines for this type of study.
Healthy adult albino rabbits, strain HC:NZW were used. The health of the animals was routinely examined for the main specific pathogens by the breeder. No vaccinations or treatment with antibiotics were perf'ormed prior to receipt of' the animals or during the acclimatization phase or study period.

Housing and feeding conditions
The rabbits were individually housed in stainless steel cages with flat rod bases or plastic cages with perforated bases, under standardized conventional conditions. Excrement trays beneath the cages contained low-dust (wood) bedding. Bedding was regularly spot-checked for contaminants at the instance of the Department of Laboratory Animal Services, and changed at least twice weekly.
Identification of animals: The rabbits were identified by individual ear marks (tattoos) and cage cards.

Acclimatization:
Prior to the initiation of the treatment the animals were kept for at least 14 days in the quarantine station of the Department of Laboratory Animal Services and monitored for diseases. During this period pooled faeces specimens were examined for Coccidia oocysts.

Animal housing conditions:
All the animals in this study were kept in one room.

Climatic conditions in animal room:
The environmental conditions were adjusted as follows:
Room temperature: 20 ± 3 °C
Humidity, relative: approx. 50 %
Light-/Dark cycle: 12 hours, artificial illumination
Air exchange rate: approx. 10 times per hour

Nutrition:
Feed: Standard diet "Ssniff K 411 (Ssniff Spezialdiäten GmbH, Soest), approx. 100 - 120 g per animal/day; once per day in the morning.
Water: Tap water; ad libitum.
Drink-water was supplied either in polycarbonate bottles containing approx. 750 ml or from automatic watering.

Weight of animals
The animals were weighed immediately before application of the test substance.

Randomization
Rabbits were randomly assigned to the respective treatment groups.

Type of coverage:

semiocclusive

Preparation of test site:

other: shorn

Vehicle:

water

Controls:

not required

Amount / concentration applied:

500 mg of the pulverized test substance

Duration of treatment / exposure:

4 hours

Observation period:

7 days

Number of animals:

3 animals

Details on study design:

Test for irritant/corrosive effects - skin

Procedure
Approximately 24 hours before the test, fur was shorn from the dorso-lateral area of the trunk (6 x 6 cm) of each of three rabbits. 500 mg of the pulverized test substance were moistened with water (to ensure good contact with the skin) and subsequently applied to a hypoallergenic patch. A further patch was moistened with water. The patches prepared in this way were placed on the opposite dorso-lateral areas of the trunk of each animal and were loosely held in place with a semiocclusive dressing for the duration of the exposure period. Thus, access by the animal to the patch and resultant ingestion/inhalation of the test substance was prevented. The treated skin area was approx. 6 cm² in size. After an exposure period of four hours, the dressing and patches were removed. The exposed skin areas were carefully washed with water without altering the existing response, or the integrity of the epidermis. The contralateral skin area not treated with test substance served as control.

Clinical observations and scoring
Dermal irritation was scored and recorded after termination of exposure at 1h, 24h, 48h, 72h and 7d . The degree of erythema/eschar formation and oedema formation was recorded as specified by DRAIZE, and any serious lesions or toxic effects other than dermal irritation were also recorded.

Irritation parameter:

erythema score

Remarks on result:

other: evaluation not possible due to the intense coloration by the test substance

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: score = 0 at any time point

Irritant / corrosive response data:

During the first 72 hours evaluation of erythema was not possible due to the intense coloration by the test substance (exposure period: 4 hrs). Other dermal reactions (e.g. edema) were not observed during this period and also no skin erythema became evident on day 7. Therefore, a significant irritant potential of the test substance to the skin is unlikely.

Interpretation of results:

GHS criteria not met

Conclusions:

Evaluation of erythema was not possible during the first 72 hours due to the intense coloration by the test substance (exposure period: 4 hrs). Edema score was = at any time point (1h, 24h, 48h, 72h and 7d).

Executive summary:

500 mg of the pulverized test substance were moistened with water (to ensure good contact with the skin) and subsequently applied to a hypoallergenic Hansamed® - patch. ( A further patch was moistened with water. The patches prepared in this way were placed on the opposite dorso-lateral areas of the trunk of each animal and were loosely held in place with a semiocclusive dressing for the duration of the exposure period. The treated skin area was approx. 6 cm² in size. After an exposure period of four hours, the dressing and patches were removed. The exposed skin areas were carefully washed with water and scored according to Draize. During the first 72 hours evaluation of erythema was not possible due to the intense coloration by the test substance (exposure period: 4 hrs). Other dermal reactions  (e.g. edema) were not observed during this period and also no skin erythema became evident on day 7. Therefore, a significant irritant potential of the test substance to the skin is unlikely.

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Principles of method if other than guideline:

After gently pulling the lower lid away from the eyeball a volume of 100 µl of the pulverized test substance - equivalent to approx. 45 mg - was placed into the conjunctival sac of one eye of each of three rabbits. The lids were then gently held together for about one second in order to limit loss of the material. The other eye remained untreated and served as control. 24 hours after instillation of the test substance the treated eye was rinsed with saline and scored according to Draize.

GLP compliance:

yes

Specific details on test material used for the study:

State of Aggregation: solid
External Appearance: yellow-brown powder
contents: 100%
pH: 6-7 (40 g/L H20)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Species and species justification
The study was conducted on rabbits an animal species recommended in the guidelines for this type of study.
Healthy adult albino rabbits, strain HC:NZW were used. The health of the animals was routinely examined for the main specific pathogens by the breeder. No vaccinations or treatment with antibiotics were perf'ormed prior to receipt of' the animals or during the acclimatization phase or study period.

Housing and feeding conditions
The rabbits were individually housed in stainless steel cages with flat rod bases or plastic cages with perforated bases, under standardized conventional conditions. Excrement trays beneath the cages contained low-dust (wood) bedding. Bedding was regularly spot-checked for contaminants at the instance of the Department of Laboratory Animal Services, and changed at least twice weekly.
Identification of animals: The rabbits were identified by individual ear marks (tattoos) and cage cards.

Acclimatization:
Prior to the initiation of the treatment the animals were kept for at least 14 days in the quarantine station of the Department of Laboratory Animal Services and monitored for diseases. During this period pooled faeces specimens were examined for Coccidia oocysts.

Animal housing conditions:
All the animals in this study were kept in one room.

Climatic conditions in animal room:
The environmental conditions were adjusted as follows:
Room temperature: 20 ± 3 °C
Humidity, relative: approx. 50 %
Light-/Dark cycle: 12 hours, artificial illumination
Air exchange rate: approx. 10 times per hour

Nutrition:
Feed: Standard diet "Ssniff K 411 (Ssniff Spezialdiäten GmbH, Soest), approx. 100 - 120 g per animal/day; once per day in the morning.
Water: Tap water; ad libitum.
Drink-water was supplied either in polycarbonate bottles containing approx. 750 ml or from automatic watering.

Weight of animals
The animals were weighed immediately before application of the test substance.

Randomization
Rabbits were randomly assigned to the respective treatment groups.

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

A volume of 100 µl of the pulverized test substance - equivalent to approx. 45 mg - was placed into the conjunctival sac of one eye.

Duration of treatment / exposure:

24 hours after instillation of the test substance the treated eye was rinsed with saline.

Observation period (in vivo):

7 days.

Number of animals or in vitro replicates:

3 animals.

Details on study design:

Procedure
After gently pulling the lower lid away from the eyeball a volume of 100 µl of the pulverized test substance - equivalent to approx. 45 mg - was placed into the conjunctival sac of one eye of each of three rabbits. The lids were then gently held together for about one second in order to limit loss of the material. The other eye remained untreated and served as control. 24 hours after instillation of the test substance the treated eye was rinsed with saline.

Clinical observations and scoring
Eye irritation was scored and recorded at 1h, 24h, 48h, 72h, 7d. The signs of cornea (opacity and area affected), iris (hyperaemia, reaction to light), conjunctivae i.e. conjunctiva of bulbus, lids, and nictitating membrane - (erythema, chemosis), and discharge were recorded as described by DRAIZE, and the aqueous humour (opacity). In addition any serious lesions or toxic effects other than ocular ones were recorded. The examinations of cornea, iris and aqueous humour were facilitated using optical instruments (e.g. hand slit-lamp). To define epithelial damage, one drop of a 1 % fluorescein solution was applied to the corneal surface 24 hours after administration of the test substance; where positive effects were recorded this was repeated at the later observation times. The eye was then rinsed with saline to remove excess and nonabsorbed fluorescein. Evaluation was performed by means of ultraviolet illumination (area) in a darkened room and diffuse white illumination.

Evaluation of results
Only effects persisting for more than 24 hours were included in the evaluation. The irritation indices / mean irritation indices were calculated for cornea (degree of opacity), iris, erythema and swelling (chemosis) of the conjunctivae.

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: score = 0 at any time point

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: score = 0 at any time point

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0.77

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: score = 0 at any time point

Irritant / corrosive response data:

Exposure of the test substance to the eye caused slight erythematous reactions of the mucous membranes. These reactions which were only transiently evident in one animal remained apparent in another animal up to 72 hours following instillation.

Interpretation of results:

GHS criteria not met

Conclusions:

Exposure of the test substance to the eye caused slight erythematous reactions (mean score = 0.77 at 24,48, and 72 hours) of the mucous membranes. These reactions which were only transiently evident in one animal remained apparent in another animal up to 72 hours following instillation.

Executive summary:

A study according to OECD guideline 405 was conducted. After gently pulling the lower lid away from the eyeball a volume of 100 µl of the pulverized test substance - equivalent to approx. 45 mg - was placed into the conjunctival sac of one eye of each of three rabbits. The lids were then gently held together for about one second in order to limit loss of the material. The other eye remained untreated and served as control. 24 hours after instillation of the test  substance the treated eye was rinsed with saline and scored according to Draize. Exposure of the test substance to the eye caused slight erythematous reactions (mean score = 0.77 at 24,48, and 72 hours) of the mucous membranes. These reactions which were only transiently evident in one animal remained apparent in another animal up to 72 hours following instillation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Bayscript Gelb BR is not irritating/corrosive to the skin and not irritating to the eyes. According to CLP classification criteria (Regulation (EC) No 1272/2008) a classification is not justified.