Reaction products of 4-amino-5-hydroxynaphthalene-2,7-disulfonic acid, coupled twice with diazotized 2-[(4-aminophenyl)sulfonyl]ethyl hydrogen sulfate, sodium salts

Endpoint summary

Description of key information

In a study which was performed to determine the skin sensitisation potential of test material in guinea pigs according to OECD Guideline 406 (Skin Sensitisation), animals did not show any reactions when tested at 5% concentration intradermal induction and at 100% concentration dermal challenge exposure.

In 1^stand 2^ndreading at 48 and 72 h respectively at 100 % no reactions were observed other than blue staining of the skin. Based on the results, the test item is concluded as not sensitizing in Pirbright white guinea pigs.

According to EU Guidelines, the test substance was not sensitizing in the guinea pig maximization test. However, literature data report on urticaria and eczema observed in workers with a high occupational exposure to reactive dyes

Consequently, it was agreed between members of the ETAD to classify the test item as skin sensitizer.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17. Nov to 18. Dec 1987

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

Data from a reliable in vivo test (non-LLNA method) conducted before the enforcement of Commission Commission Regulation (EU) 2017/706 of 19 April 2017 amending Annex VII to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) as regards skin sensitisation are available.

Species:

guinea pig

Strain:

Pirbright-Hartley

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Hoechst AG
- Weight at study initiation: 240 to 334 g
- Housing: group-housing (5/cage)
- Diet: ERKA Nr 8300 ad libitum
- Water: tap water ad libitum
- Acclimation period: ca. 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25°C
- Humidity (%): 30 to 70%
- Photoperiod (hrs dark / hrs light): 12/12


IN-LIFE DATES: From: 17. Nov To: 18. Dec 1987

Route:

intradermal and epicutaneous

Vehicle:

physiological saline

Concentration / amount:

Intradermal: 5 %
dermal induction: 100 %
dermal challenge: 100 %

Route:

epicutaneous, occlusive

Vehicle:

physiological saline

Concentration / amount:

Intradermal: 5 %
dermal induction: 100 %
dermal challenge: 100 %

No. of animals per dose:

Determination of primary not irritating concentration: 6
Determination of intradermal tolerability: 3
Sentinel group: 5
Control group: 5
Treatment group: 10

Positive control substance(s):

yes

Reading:

1st reading

Hours after challenge:

48

Group:

test chemical

Dose level:

100 %

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

blue staining of skin

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 100% challenge. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: blue staining of skin.

Reading:

2nd reading

Hours after challenge:

72

Group:

test chemical

Dose level:

100 %

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

blue staining of skin

Remarks on result:

other: Reading: 2nd reading. Hours after challenge: 72.0. Group: test group. Dose level: 100 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: blue staining of skin.

Reading:

1st reading

Hours after challenge:

48

Group:

negative control

Dose level:

100 %

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

blue staining

Remarks on result:

no indication of skin sensitisation

Reading:

2nd reading

Hours after challenge:

72

Group:

negative control

Dose level:

100 %

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

blue staining

Remarks on result:

no indication of skin sensitisation

Reading:

1st reading

Group:

positive control

Remarks on result:

not measured/tested

Interpretation of results:

not sensitising

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

The test item is not sensitizing in pirbright white guinea pigs. Classification: not sensitizing

Executive summary:

A key study was performed to determine the skin sensitisation potential of test material in rabbits according to OECD Guideline 406 (Skin Sensitisation).

The test was conducted with the following concentrations

Intradermal: 5 %

Dermal induction: 100 %

Dermal challenge: 100 %

In 1^st and 2^nd reading at 48 and 72 h respectively at 100 % no reactions were observed other than blue staining of the skin.

Based on the results, the test item is concluded as not sensitizing in pirbright white guinea pigs.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Respiratory sensitisation

Link to relevant study records

Reference

Endpoint:

respiratory sensitisation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11. Aug to 21. Oct. 1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Principles of method if other than guideline:

According to the European Discussion Group of Inhalation Toxicologists (EDIT) referring to Botham P.A., Rattraya N.J., Woodcocka D.R., Walsha S.T. and Hexta P.M. "The induction of respiratory allergy in guinea-pigs following intradermal injection of trimellitic anhydride: a comparison with the response to 2,4-dinitrochlorobenzene ", Toxicology Letters, Volume 47, Issue 1, April 1989, Pages 25-39

GLP compliance:

yes

Species:

guinea pig

Strain:

other: Pirbright-White (HOE DHPK (SPFLac))

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Hoechst AG
- Age at study initiation: 3 to 5 weeks
- Weight at study initiation (mean): males: 250 g; females: 243 g
- Housing: groups of 4 animals
- Diet: Altromin 3112 ad libitum
- Water: tap ad libitum
- Acclimation period: at least 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 24
- Humidity (%): 30 to 70
- Air changes (per hr): -
- Photoperiod (hrs dark / hrs light): 12/12


IN-LIFE DATES: From: 11-Aug-1993 To: 21-Oct-1993

Route of induction exposure:

intradermal

Route of challenge exposure:

inhalation

Vehicle:

other: intradermal: NaCl

Concentration:

males: 150, 210 mg/m³
females: 140, 180 mg/m³

No. of animals per dose:

Determination of the tolerance of intradermal injections: 2 animals
Determination of the primary non-irritant concentration (inhalation): 4 animais
Material Control Group: 8 animals
Test Group: 8 animals

Details on study design:

Animals of the test groups are treated intradermally with the test substance: 0.1 mL 1 %, 5 %, 30 %
Animals of the material control groups are treated with the vehicle only.
Determination of the primary non irritant aerosol concentration: 30 to 200 mg/m³
All animals are challenged by inhalation after three weeks using a primary non irritating concentration of the test substance.
Allergic reactions in the test groups are assessed by changes of lung function parameters compared to the material control groups.

Challenge controls:

yes

Positive control substance(s):

not specified

Negative control substance(s):

not specified

Results:

Determination of the tolerance of intradermal injections:
Intradermal injection of 30 % of the test item in physiological saline caused encrustations and beginning necrosis at the application sites. Slight induration was observed after injection of the 5 % formulation. The injection sites treated with 1 % of the test item showed no signs of irritation. Based on these results a 5 % solution was chosen for intradermal induction at day 1.

Determination of the primary non-irritant aerosol concentration:
A slight increase in respiratory rate occurred during exposure to approx. 200 mg/mg air. No marked changes in respiratory rate were observed during exposure to approx. 160 mg test item/m³. Therefore this concentration was chosen for inhalation exposure at challenge day 22.

Body weight gain and clinical signs:
The intradermal injections caused encrustations and indurations of the injection sites up to day 8 at the study. Body weight gains were not impaired.

Challenge treatment
– Lung function parameters
Questionable up to slight changes in the respiratory pattern were observed in all animals after onset of exposure. No significant differences were present between the animals of the material control groups and the animals of the test groups.
Based on the results of the present study the test item did not cause a significant allergic response after intradermal induction and respiratory challenge.

- Clinical signs of intoxication
No clinical signs of intoxication were observed.

- Autopsy findings
Autopsy of the animals revealed red patches on the lungs in one animal of each group, respectively.

Positive control results:

not examined

Negative control results:

true negative control substances have not been examined

Interpretation of results:

not sensitising

Conclusions:

Based on the results of the present study the test item did not cause a significant allergic response after intradermal induction and respiratory challenge.

Executive summary:

A GLP compliant study was conducted according to the European Discussion Group of Inhalation Toxicologists (EDIT) referring to Botham P.A., Rattraya N.J., Woodcocka D.R., Walsha S.T. and Hexta P.M. "The induction of respiratory allergy in guinea-pigs following intradermal injection of trimellitic anhydride: a comparison with the response to 2,4-dinitrochlorobenzene ", Toxicology Letters, Volume 47, Issue 1, April 1989, Pages 25-39

The guinea pigs were placed individually in cylindrical plastic tubes connected with plethysmographs and exposed to specified aerosol concentrations. The plastic tubes leading into the exposure chamber were arranged in such a way that only the noses of the animals were inside the chamber.

The test item was tested for respiratory sensitization in Pirbright White guinea pigs. Induction was carried out by intradermal injection of 0.1 ml the test item (5% solution in physiological saline) at day 1. Animals were challenged at day 22 with approx. 140 - 210 mg of test item / m3 air.

Evaluation of the particle size distribution of the test item aerosol revealed a mass median aerodynamic diameter (MMAD) of 1.8 micrometers and a geometric standard deviation of 1.9. The respiratory pattern changed slightly in some animals during challenge on day 22, but there were no marked differences between the animals induced with the test item and control animals, which received isotonic saline only on the day of induction.

Based on the results of the present study the test item did not cause a significant allergic response after intradermal induction and respiratory challenge.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

According to EU Guidelines, the test substance was not sensitizing in an inhalative guinea pig sensitization test performed according to the European Discussion Group of Inhalation Toxicologists (EDIT) (referring to Botham P.A., Rattraya N.J., Woodcocka D.R., Walsha S.T. and Hexta P.M. "The induction of respiratory allergy in guinea-pigs following intradermal injection of trimellitic anhydride: a comparison with the response to 2,4-dinitrochlorobenzene ", Toxicology Letters, Volume 47, Issue 1, April 1989, Pages 25-39).

However, literature data report on symptoms of respiratory allergy observed in workers with a high occupational exposure to reactive dyes.

Consequently, it was agreed between members of the ETAD to classify the test substance as respiratory sensitizer.

Justification for classification or non-classification

Based on the finding of the skin and respiratory sensitization studies in vivo, the test substance does not need to be classified according to Directive 67/548/EEC and according to EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.

Nevertheless, based on available literature data on workers being sensitzed upon high occupational exposure to reactive dyes it was agreed between members of ETAD to classify and label Reactive Black 5 as skin and respiratory sensitizer (Guidance to ETAD Member Companies on the implementation of the Code of Ethics, published Basle 03/2001 and revised 04/2008).