trisodium 4-hydroxy-6-(sulfonatomethylamino)-5-(2-(2-sulfatoethylsulfonyl)phenylazo)naphthalene-2-sulfonate

Endpoint summary

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:

From January 12, 1998 to January 15, 1998

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)

Version / remarks:

adopted 1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Remarks:

Chbb:NZW(SPF)

Details on test animals or test system and environmental conditions:

Source: Boehringer Ingelheim Pharma KG, Birkendorferstrasse 65, D-88397 Biberach/Riss
Number of animals: 1 male, 2 females
Age at start of treatment: 15 weeks
Identification: by unique cage number and corresponding ear number
Acclimatization: Four days under test conditions after health examination.

Type of coverage:

semiocclusive

Preparation of test site:

other: The dorsal fur was clipped with an electric clipper.

Vehicle:

other: The test substance was moistened with bi-distilled water before application.

Amount / concentration applied:

Soli 0.5 g /animal (left side only)

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

3

Details on study design:

Approximately three days before treatment the dorsal fur was clipped with an electric clipper, exposing an area of approximately 100m2 (10cm x 10 cm). The skin of the animals was examined approximately 24 hours before treatment and if necessary regrown fur was again clipped.

On the day of treatment the test substance was applied to approximately 6cm2 of the intact skin of the clipped area. It was covered with a 25 cm x 25 cm patch of surgical gauze and the gauze was covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and anchored with tape.

The duration of the treatment was 4 hours. Then the dressing was removed and the skinwas flushed with lukewarm tap water to clean the applicationsite so that any reactions (erythema) were clearly visible at that time.

Observations
Mortality/ viability: daily
Clinical signs: daily during the observation period
Body weights: a start of acclimatization, in the first day of application and at termination of observation

Irritation scores
The skin reaction was assessed according to the numerical scoring system listed in the EEC Comission Directive 92/69/EEC, 1992 at approximately 1, 24, 48 and 72 hours after the removal of the dressing gaze patch and test substance.

Pathology
No necropsy was performed in the animals acrifficed at termination of observation. All rabbits were sacrifices by an intravenous injection of Narcoren into the ear vein of at least 1 ml/kg body weight.

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:

Application of the test substance to healthy intact rabbit skin resulted in a primary irritation score of 0.00. Local signs consisted of grade 0.00 edema (mean values from 24 to 72 hours) and grade 0.00 erythema (mean values from 72 hours).
Violet-red staining by the test substance o the treated skin was observed.
No corrosive effect and no irreversible alterations of the treated skin were observed.

Other effects:

No clinical signs of systemic toxicity were observed in the animals during the test and observation period, and no mortality occurred. The body weight of all rabbits was considered to be within the normal range of variability.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the study conditions, the test substance was considered to be not irritating to rabbit skin.

Executive summary:

A study was conducted to determine the skin irritation potential of the test substance according to OECD Guideline 404 and EU Method B.4, in compliance with GLP. The primary skin irritation potential of the test substance was investigated by topical application of 0.5 g to 6 cm2 of intact dorsal skin of each of three young adult New Zealand White rabbits. The duration of the treatment was 4 h. The scoring of skin reactions was performed at 1, 24, 48 and 72 h after removal of the dressing. The scores of each animal at 24, 48 and 72 h were used in calculating the respective mean values for each type of lesion. The primary irritation score was calculated by totalling the individual cumulative scores at 24, 48, 72 h and then dividing by the number of data points. The primary irritation score was 0.00 (max. 8.0). Local signs consisted of grade 0.00 oedema (mean values from 24 to 72 h) and grade 0.00 erythema (mean values from 72 h). The test substance caused violet-red staining of the treated skin. No corrosive effects were noted on the treated skin of any animal at any measuring interval. Under the study conditions, the test substance was considered to be not irritating to rabbit skin (Braun, 1998)..

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:

From January 19, 1998 to January 22, 1998

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)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Remarks:

Chbb: NZW (SPF)

Details on test animals or tissues and environmental conditions:

Source: Boehringer Ingelheim Pharma KG, Birkendorferstrasse 65, D-88397 Biberach/Riss
Number of animals: 1 male, 2 females
Age at start of treatment: 15 weeks
Identification: by unique cage number and corresponding ear number
Acclimatization: Four days under test conditions after health examination.
Room temperature/ relative humidity: air conditioned with 10-15 air changes per hour, with target ranges for room temperature 20 ±3 °C and of 40 – 70 % humidity.
Day/night rhythm: 12h/ 12h
Type of cage: single, stainless steel wire mesh cages with automatic cleaning system equipped with feed hoppers, drinking water bowls and wood for gnawing.
Bedding: no bedding in the cages, wood shavings in the waste trays
Diet: Pelleted standard Kliba-3410 rabbit maintenance diet ad libitum
Water: tap water, from Itingen, ad libitum in water bowls

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

0.1 g

Duration of treatment / exposure:

24 hours

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3

Details on study design:

The eyes of the animals were examined one day prior to test article administration. On the day of the treatment the test substance (undiluted) 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 test substance. The right eye remained untreated and served as the reference control. The treatment eyes were rinsed gently with physiological saline approximately 24 hours after administration.

Observations
Mortality/ viability: daily
Clinical signs: daily during the observation period
Body weights: a start of acclimatization, on the first day of application and at termination of observation

The ocular reaction was assessed according to the numerical according system listed in EEC Commission Directive 92/69/EEC, 1992 at approximately 1, 24, 48 and 72 hours after administration.

Pathology
No necropsy was performed in the animals sacrificed at termination of observation. All rabbits were sacrifices by an intravenous injection of Narcoren into the ear vein of at least 1 ml/kg body weight.

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

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Application of the test substance to healthy intact rabbit conjunctivae resulted in a primary irritation score of 0.00. Slight swelling of the conjunctivae and moderate watery discharge were noted in all animals once hour after application. All signs of irritation were reversible after 24 hours.
Reversible light violet staining of the sclera and conjunctivae by the test substance was observed. The corneas were not affected. No corrosion of the cornea was observed atany of the reading times.

Other effects:

No clinical signs of systemic toxicity were observed in the animals during the test and observation period, and no mortality occurred.
The body weight of all rabbits was considered to be within the normal range of variability.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the study conditions, the test substance was considered to be non-irritant to the rabbit eye.

Executive summary:

A study was conducted to determine the potential of the test substance to cause damage to conjunctiva, cornea or iris after a single exposure of about 24 h according to OECD 405 Guideline and EU Method B.5, in compliance with GLP. The primary irritation potential was investigated by instillation of 0.1 g into one eye of each of three young adult New Zealand White rabbits. The treated eyes were rinsed gently with physiological saline approximately 24 h after administration. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 h after test substance application. The scores of each animal at 24, 48 and 72 h were used in calculating the respective mean values for each type of lesion. The primary irritation score was calculated by totalling the individual cumulative scores at 24, 48 and 72 h and then dividing the resulting total by the number of data points. The primary irritation score was 0.00 (max. 13). Swelling of the conjunctivae and watery discharge were noted at all animals 1 h after application. All signs of irritation were reversible after 24 h. Reversible light violet staining of the sclera and conjunctivae by the test substance was observed. The corneas were not affected. No corrosion was observed at any of the measuring intervals. Under the study conditions, the test substance was considered to be non-irritant to the rabbit eye (Braun, 1998).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin

A study was conducted to determine the skin irritation potential of the test substance according to OECD Guideline 404 and EU Method B.4, in compliance with GLP. The primary skin irritation potential of the test substance was investigated by topical application of 0.5 g to 6 cm2 of intact dorsal skin of each of three young adult New Zealand White rabbits. The duration of the treatment was 4 h. The scoring of skin reactions was performed at 1, 24, 48 and 72 h after removal of the dressing. The scores of each animal at 24, 48 and 72 h were used in calculating the respective mean values for each type of lesion. The primary irritation score was calculated by totalling the individual cumulative scores at 24, 48, 72 h and then dividing by the number of data points. The primary irritation score was 0.00 (max. 8.0). Local signs consisted of grade 0.00 oedema (mean values from 24 to 72 h) and grade 0.00 erythema (mean values from 72 h). The test substance caused violet-red staining of the treated skin. No corrosive effects were noted on the treated skin of any animal at any measuring interval. Under the study conditions, the test substance was considered to be not irritating to rabbit skin (Braun, 1998).

Eyes

A study was conducted to determine the potential of the test substance to cause damage to conjunctiva, cornea or iris after a single exposure of about 24 h according to OECD 405 Guideline and EU Method B.5, in compliance with GLP. The primary irritation potential was investigated by instillation of 0.1 g into one eye of each of three young adult New Zealand White rabbits. The treated eyes were rinsed gently with physiological saline approximately 24 h after administration. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 h after test substance application. The scores of each animal at 24, 48 and 72 h were used in calculating the respective mean values for each type of lesion. The primary irritation score was calculated by totalling the individual cumulative scores at 24, 48 and 72 h and then dividing the resulting total by the number of data points. The primary irritation score was 0.00 (max. 13). Swelling of the conjunctivae and watery discharge were noted at all animals 1 h after application. All signs of irritation were reversible after 24 h. Reversible light violet staining of the sclera and conjunctivae by the test substance was observed. The corneas were not affected. No corrosion was observed at any of the measuring intervals. Under the study conditions, the test substance was considered to be non-irritant to the rabbit eye (Braun, 1998).

Justification for classification or non-classification

Based on the results of in vivo skin and eye irritation testing in rabbits, the substance does not require classification for these endpoints according to CLP (EC 1272/2008) criteria.