Tetrasodium 4-amino-6-[[2,5-dimethoxy-4-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]azo]-5-hydroxy-3-[[4-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]azo]naphthalene-2,7-disulphonate

Respiratory sensitisation

Administrative data

Endpoint:

respiratory sensitisation: in vivo

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

11. Aug to 21. Oct. 1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1993

Report date:

1993

Materials and methods

Principles of method if other than guideline:

According to the European Discussion Group of Inhalation Toxicologists (EDIT) referring to Botham P.A., Rattray N.J., Woodcock D.R., Walsh S.T. and Hext 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

Test material

Test material information

Constituent 1

Reference substance name:

Reactive Black 5

IUPAC Name:

Reactive Black 5

Details on test material:

- Name of test material (as cited in study report): Remazol Schwarz B

Test animals

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

Test system

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 and discussion

Results:

Determination of the tolerance of intradermal injections:
Intradermal injection of 30% Remazol-Schwarz B 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% Remazol - Schwarz B 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 Remazol-Schwarz B/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 Remazol-Schwarz B 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:

-

Negative control results:

-

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Conclusions:

Based on the results of the present study Remazol-Schwarz B did not cause a significant allergic response after intradermal induction and respiratory challenge.

Executive summary:

Remazol-Schwarz B was tested for respiratory sensitization in Pirbright White guinea pigs. Induction was carried out by intradermal injection of 0.1 ml Remazol-Schwarz B (5% solution in physiological saline) at day 1. Animals were challenged at day 22 with approx. 140 - 210 mg Remazol-Schwarz B / m3 air. Evaluation of the particle size distribution of Remazol-Schwarz B 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 Remazol-Schwarz B and control animals, which received isotonic saline only on the day of induction. Based on the results of the present study Remazol-Schwarz B did not cause a significant allergic response after intradermal induction and respiratory challenge.