Reaction products of diazotised 2-aminophenol-4-sulfonamide coupled with 7-amino-4-hydroxynaphthalene-2-sulfonic acid, subsequently diazotised and coupled with 3-oxo-N-phenylbutanamide and 3 aminophenol, chelated with chromium (3+), potassium salts

Endpoint summary

Description of key information

skin irritation, rabbit, semiocclusive, not irritating

eye irritation, rabbits, irritating

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

other: read across from analogue substance

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Kleintierfarm Madörin AG, 4414 Füllingsdorf, CH
- Age at study initiation: 14 to 15 weeks
- Weight at study initiation: 2.6 to 2.7 kg
- Housing: individually in stainless steel cages equipped with an automatic cleaning and drinking system (Dipl. Ing. W. Ehret Gmbh, Versuchstiertechnik, 7830 Emmendingen, Germany)
- Diet: ad libitum; pelleted standard Kliba 341, batch 30/87 rabbit maintenance diet ("Kliba", Klingentalmühle AG, 4303 Kaiseraugst, CH) (analytical test report demonstrates suitability)
- Water: ad libitum; community tap water from Itingen (bacteriological assay and chemical water analysis demonstrated suitability)
- Acclimation period: 4 days

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

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

water

Controls:

no

Amount / concentration applied:

- Amount applied: 0.5 g solid

Duration of treatment / exposure:

4h

Observation period:

24, 48 and 72h

Number of animals:

3

Details on study design:

TEST SITE
- Shaved area: 100 cm² (10 cm x 10 cm)
- Area of exposure: 9 cm² (3 cm x 3 cm)
- Location: dorsal
- Type of wrap if used: surgical gauze

REMOVAL OF TEST SUBSTANCE
- Semi-occlusive dressing
- Test item removed and washed with lukewarm tap water 4 hours after application

OBSERVATION TIME POINTS
- 1, 24, 48 and 72 hours after application

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

ca. 0

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

ca. 0

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

ca. 0

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

ca. 0

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

ca. 0

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

ca. 0

Irritant / corrosive response data:

No irritative response was detected based on erythem and edema score
No corrosion was observed

Other effects:

In the area of application slight black staining of the treated skin by pigment or colouring of the test item was observed after 1 hour in 2 rabbits. No rabbits demonstrated discolouration after 24 hours.

BODY WEIGHTS
- The body weight gain of all rabbits was similar.

TOXIC SYMPTOMS / MORTALITY
- No acute toxic symptoms were observed in the animals during the test period, and no mortality occurred.

NECROPSY
- Due to the results obtained, no macroscopic organ examination was indicated.

Interpretation of results:

other: not classified under Regulation 1272/2008

Conclusions:

The substance was tested for skin irritation/corrosion following OECD 404. Under the experimental conditions the substance did not show any skin irritant or corrosive properties.

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:

other: read across from analogue substance

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

performed on analogue substance

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Kleintierfarm Madörin AG, 4414 Füllinsdorf, CH
- Age at study initiation: 14 to 15 weeks
- Weight at study initiation: 2.5 to 2.6 kg
- Housing: individually in stainless steel cages equipped with an automatic cleaning and drinking system (Dipl. Ing. W. Ehret Gmbh, Versuchstiertechnik, 7830 Emmendingen, Germany)
- Diet: ad libitum; pelleted standard Kliba 341, batch 30/87 rabbit maintenance diet ("Kliba", Klingentalmühle AG, 4303 Kaiseraugst, CH) (analytical test report demonstrates suitability)
- Water: ad libitum; community tap water from Itingen (bacteriological assay and chemical water analysis demonstrated suitability)
- Acclimation period: 4 days under laboratory conditions after veterinary examination.

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

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

100 mg

Duration of treatment / exposure:

single application

Observation period (in vivo):

14 days

Number of animals or in vitro replicates:

3

Details on study design:

TREATMENT
- The test item was placed in the conjunctival sacs of the left eye of each animal after gently pulling the lower lid away from the eye ball. The lids were then gently held together for about 1 second to prevent loss of the test item.

REMOVAL OF TEST SUBSTANCE :
- Washing: no washing

TOOL USED TO ASSESS SCORE:
- Eye examinations were made with a slit-lamp 30 SL (C. Zelss AG, Zürich, CH) and a "Varta Cliptrix" diagnostic-lamp (A. Riegger, Basel, CH)

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Time point:

24/48/72 h

Score:

ca. 1

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

24/48/72 h

Score:

ca. 1.7

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Time point:

24/48/72 h

Score:

ca. 0

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

ca. 0.7

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

ca. 1.3

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

ca. 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

ca. 1

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

ca. 1

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

ca. 1

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

ca. 0

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

ca. 0

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

ca. 0

Irritant / corrosive response data:

only slight cornea opacity was observed within 1h to 7 days

Other effects:

TOXICITY
- No acute toxic symptoms were observed in the animals during the test period.

MORTALITY
- No mortality occurred.

COLOURATION
- In the area of applicatlon a slight black to black staining of the cornea, lidhairs and conjunctivae by pigment or colouring of the test item was observed.

BODY WEIGHTS
- The body weight gains of all rabbits was similar.

NECROPSY
- No macroscopic organ examination was performed.

Interpretation of results:

other: classified as H319 Eye Irritation 2

Conclusions:

The substance was tested for eye irriation properties following OECD 405. Under the experimental conditions the cornal opacity resulted equal to 1 for 3/3 animals which classified the substance as H319: Causes serious eye irriation.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

The analogue substance was tested in two in-vivo studies for skin and eye irritation folloiwng EU Method B4 and B5 (1989).

Justification for classification or non-classification

According to the CLP Regulation (EC n. 1272/2008) the following classifications apply:

 

Skin irritant cat2:

1) 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

2)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; o

3) 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.

 

 

Eye irritant Cat 2:

at least in 2 of 3 tested animals, a positive response of:

1)     corneal opacity ≥ 1 and/or

2)     iritis ≥ 1, and/or

3)      conjunctival redness ≥ 2 and/or

4)      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 the results of the in vivo eye irritation, the substance is classified as H319, Eye Irritation 2.