Benzoic acid, 4-[2-(2-hydroxy-6-sulfo-1-naphthalenyl)diazenyl]-, strontium salt (2:1)

Endpoint summary

Description of key information

Skin Irritation 
in vivo (rabbit): not irritant; OECD Guideline 404, GLP, Safepharm Laboratories Ltd, 1235/004, 1998. 
Eye Irritation: 
in vivo (rabbit): not irritant; OECD Guideline 405, GLP, Safepharm Laboratories Ltd, 1235/005, 1998

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:

other: GLP - Guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

(adopted 17 July 1992)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: David Percival Ltd., Moston, Sandbach, Cheshire, UK
- Age at study initiation: twelve to sixteen weeks old
- Weight at study initiation: 2.53 - 2.74 kg
- Housing: individually in suspended metal cages
- Diet: STANRAB SQC Rabbit Diet, Special Diets Services Ltd, Witham, Essex, UK, ad libitum
- Water: mains drinking water, ad libitum
- Acclimation period: five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 21
- Humidity (%): 65 - 74. On one occasion the relative humidity was above the limit specified in the study protocol (70%). This deviation was considered not to affect the purpose or integrity of the study.
- Air changes (per hr): approximately 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12 / 12

Type of coverage:

semiocclusive

Preparation of test site:

other: clipped

Vehicle:

water

Controls:

not specified

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g
- Concentration (if solution): 0.5 g moistened with 0.7 ml of distilled water
- The test material was introduced under a cotton gauze patch and placed in position on the shorn skin.

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

3

Details on study design:

TEST SITE
- On the day before the test each rabbit was clipped free of fur from the dorsal flank area using veterinary clippers. Only animals with a healthy intact epidermis by gross observation were selected for the study.
- Area of exposure: back
- coverage: The test material was introduced under a 2.5 cm x 2.5 cm cotton gauze patch and placed in position on the shorn skin. The patch was secured in position with a strip of surgical adhesive tape (BLENDERM: approximate size 2.5 cm x 4.0 cm).
- Type of wrap if used: To prevent the animals interfering with the patches, the trunk of each rabbit was wrapped in an elasticated corset (TUBIGRIP) and the animals were returned to their cages for the duration of the exposure period.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The corset and patches were removed from each animal and any residual test material removed by gentle swabbing with cotton wool soaked in 74% Industrial Methylated Spirits.
- Time after start of exposure: Four hours after application

SCORING SYSTEM: Approximately one hour following the removal of the patches, and 24, 48 and 72 hours later, the test sites were examined for evidence of primary irritation and scored according to the scale from Draize J.H. (1959) Association of Food and Drug Officials of the United States, Austin, Texas, "The Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics".

Irritation parameter:

erythema score

Basis:

mean

Remarks:

of three animals

Time point:

other: 24, 48, 72 hours

Score:

0

Max. score:

4

Reversibility:

other: no effects

Remarks on result:

other: Orange-coloured staining caused by the test material was noted at all treated skin sites throughout the study but did not affect evaluation of the skin responses.

Irritation parameter:

edema score

Basis:

mean

Remarks:

of three animals

Time point:

other: 24, 48, 72 hours

Score:

0

Max. score:

4

Reversibility:

other: no effects

Other effects:

No other effects were observed.

Interpretation of results:

GHS criteria not met

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:

other: GLP - Guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

(adopted 24 February 1987)

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

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: David Percival Ltd., Moston, Sandbach, Cheshire, UK
- Age at study initiation: twelve to sixteen weeks old
- Weight at study initiation: 2.63 - 2.79 kg
- Housing: individually in suspended metal cages
- Diet: STANRAB SQC Rabbit Diet, Special Diets Services Ltd, Witham, Essex, UK, ad libitum
- Water: mains drinking water, ad libitum
- Acclimation period: five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 21
- Humidity (%): 66 - 74. On one occasion the relative humidity was above the limit specified in the study protocol (70%). This deviation was considered not to affect the purpose or integrity of the study.
- Air changes (per hr): approximately 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12 / 12

Vehicle:

unchanged (no vehicle)

Controls:

other: The left eye remained untreated and was used for control purposes.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): A volume of 0.1 ml of the test material, which was found to weigh approximately 24 mg (as measured by gently compacting the required volume into an adapted syringe) was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were held together for about one second immediately after application, to prevent loss of the test material, and then released.

Duration of treatment / exposure:

One exposure, no washing out.

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

One rabbit was initially treated. After consideration of the ocular responses produced in the first treated animal, two additional animals were treated.

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: not done

ASSESSMENT:
- Immediately after administration of the test material, an assessment of the initial pain reaction was made.
- Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment.

SCORING SYSTEM:
- According to the numerical evaluation given in Appendix I, (from Draize JH (1977) "Dermal and Eye Toxicity Tests" In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington DC p.48 to 49).

TOOL USED TO ASSESS SCORE:
- Examination of the eye was facilitated by use of the light source from a standard ophthalmoscope.

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

of 3 animals

Time point:

other: 24, 48 and 72 hours

Score:

0

Max. score:

4

Reversibility:

other: no effects

Irritation parameter:

iris score

Basis:

mean

Remarks:

of 3 animals

Time point:

other: 24, 48 and 72 hours

Score:

0

Max. score:

2

Reversibility:

other: no effects

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

of 3 animals

Time point:

other: 24, 48 and 72 hours

Score:

0

Max. score:

3

Reversibility:

other: no effects

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of 3 animals

Time point:

other: 24, 48 and 72 hours

Score:

0

Max. score:

4

Reversibility:

other: no effects

Irritant / corrosive response data:

- No corneal or iridial effects were noted.
- Minimal conjunctival irritation (score 1) was noted in all treated eyes at the 1-hour observation.
- No signs of ocular irritation were noted at the 24-hour observation.

Other effects:

The fur around all treated eyes was stained orange throughout the study.

Interpretation of results:

GHS criteria not met

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

There are valid in vivo data available for the assessment of the skin and eye irritation potential of the test item.

Skin Irritation / Corrosion

The key study, a primary skin irritation study, was performed according to GLP and OECD Guideline 404 (Acute Dermal Irritation/Corrosion) with 0.5 g of the moistened (distilled water) test item (analytical purity unknown) applied for 4 hours to the clipped skin of each of three 12 to 14 weeks old New Zealand White rabbits under semi-occlusive conditions (Safepharm, 1235/004, 1998). The scoring of skin reactions was performed 1, 24, 48 and 72 hours, as well as 7, 10 and 14 days after removal of the test item according to the Draize classification scheme. No signs of skin irritation or skin corrosion were seen. No other signs of intoxication were observed.  

 

Eye Irritation

In the key eye irritation study, conducted according OECD Guideline 405 (Acute Eye Irritation/Corrosion), 0.1 ml of the undiluted test article (analytical purity unknown) was instilled into the non-irrigated eye of each of three 12 to 14 weeks old New Zeeland White rabbits (3 males) (Safepharm, 1235/005, 1998). Eyes were not washed after instillation. Irritation was scored 1, 24, 48 and 72 hours after treatment according to the Draize classification scheme. No signs of eye irritation were seen in treated eyes 24, 48 and 72 hours after treatment. No other signs of intoxication were observed.

Justification for classification or non-classification

Classification, Labeling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for the purpose of classification under Regulation (EC) No.1272/2008. No indication of skin or eye irritation was observed in experimental animals (OECD 404 and 405). Based on the data, classification for skin and eye irritation is not warranted under Regulation (EC) No.1272/2008.