Reaction mass of bis[3-[[4-[benzylmethylamino]phenyl]azo]-1,4-dimethyl-1H-1,2,4-triazolium] tetrachlorozincate(2-) and bis[5-[[4-[benzylmethylamino]phenyl]azo]-1,4-dimethyl-1H-1,2,4-triazolium] tetrachlorozincate(2-)

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From July 16, 1979 to September 6, 1979

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1979

Report date:

1979

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: "Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959), the US Association of Food and Drug Officials (AFDO).

Deviations:

not specified

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: Maurer Optimization test in the guinea-pig

Version / remarks:

- Maurer Th., Thomann P., Weirich E.G., Hess R. - The Optimization test in the guinea-pig. Agents & Actions 5 (2), 174-179, 1975.
- Predictive evaluation in animals of the contact allergenic potential of medically important substances. Contact Dermatitis 4, 321-333, 1978. Contact Dermatitis 5, 1-10, 1979.

GLP compliance:

no

Type of study:

Maurer optimisation test

Justification for non-LLNA method:

Study predates LLNA method

Test material

Test material information

Constituent 1

Reference substance name:

Bis[5-[[4-[benzylmethylamino]phenyl]azo]-1,4-dimethyl-1H-1,2,4-triazolium] tetrachlorozincate(2-)

EC Number:

281-692-3

EC Name:

Bis[5-[[4-[benzylmethylamino]phenyl]azo]-1,4-dimethyl-1H-1,2,4-triazolium] tetrachlorozincate(2-)

Cas Number:

84012-52-2

Molecular formula:

C36H42Cl4N12Zn

IUPAC Name:

bis[5-({4-[benzyl(methyl)amino]phenyl}diazenyl)-1,4-dimethyl-4,5-dihydro-1H-1,2,4-triazol-1-ium] tetrachlorozincate(2-)

Constituent 2

Reference substance name:

Bis[3-[[4-[benzylmethylamino]phenyl]azo]-1,4-dimethyl-1H-1,2,4-triazolium] tetrachlorozincate(2-)

EC Number:

281-650-4

EC Name:

Bis[3-[[4-[benzylmethylamino]phenyl]azo]-1,4-dimethyl-1H-1,2,4-triazolium] tetrachlorozincate(2-)

Cas Number:

84000-94-2

Molecular formula:

C36H42Cl4N12Zn

IUPAC Name:

bis(3-[(E)-2-{4-[benzyl(methyl)amino]phenyl}diazen-1-yl]-1,4-dimethyl-1H-1,2,4-triazol-4-ium); tetrachlorozincdiuide

Test material form:

solid: particulate/powder

In vivo test system

Test animals

Species:

guinea pig

Strain:

other:

Remarks:

Pirbright-White

Sex:

male/female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source:Ivanovas, Kissleg (later Charles River Kissleg), Germany
- Age at study initiation: ~10 weeks old
- Weight at study initiation: 375 to 485 g
- Housing: housed individually in Macrolon cages, type 3, assigned to the different groups by means of random numbers.
- Diet : The animals received ad libitum standard guinea pig pellets - NAFAG, N°. 830 supplemented with fresh carrots.
- Water: ad libitum
- Acclimation period: 10 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 1°C
- Humidity (%): 55± 5%
- Photoperiod (hrs dark / hrs light): 10 hours light cycle day

Study design: in vivo (non-LLNA)

Induction

Route:

intradermal

Vehicle:

physiological saline

Concentration / amount:

10 * 0.1 ml of 0.1 % of test substance every other day (except weekends)
During the second and third week of the induction period the test material was incorporated in a mixture of the normal vehicle with complete Bacto Adjuvant (Freund's complete Adjuvant) (vehicle : adjuvant =1 : 1).

Day(s)/duration:

Days 1, 3, 5, 9, 11, 13, 17, 19, 21, 25

Challenge

Challenge 1

No.:

#1

Route:

intradermal

Vehicle:

physiological saline

Remarks:

complete Bacto Adjuvant

Concentration / amount:

0.1 ml of a freshly prepared 0.1 % solution

Day(s)/duration:

14 days after last induction injection

Challenge 2

No.:

#2

Route:

epicutaneous, occlusive

Vehicle:

other: vaseline

Concentration / amount:

3 % of test item in vaseline

Day(s)/duration:

10 days after intradermal challenge for 24 hours

Adequacy of challenge:

highest non-irritant concentration

No. of animals per dose:

10 per sex per dose

Details on study design:

MAIN STUDY
A. INDUCTION EXPOSURE INTRADERMAL
- No. of exposures: the animals received one injection every second day (except weekends) to a total of 10 intracutaneous injections of a freshly prepared 0.1 % suspension of test item in physiological saline. During the second and third week of the induction period the test material was incorporated in a mixture of the normal vehicle with complete Bacto Adjuvant (vehicle : adjuvant = 1 : 1)
- Test groups: 10 males and 10 females guinea pigs
- Control group: 10 males and 10 females guinea pigs, one control group was treated with the vehicle alone
- Site: shaven skin of the right flank (only day 1) and the back
- Frequency of applications: every second day
- Duration: 10 intracutaneous injections (approximately 25 days)
- Concentrations: 0.1 % solution of test substance


B. CHALLENGE EXPOSURE
INTRADERMAL
- No. of exposures: single injection of 0.1 ml
- Day(s) of challenge: Fourteen days after the last induction injection
- Exposure period: single injection
- Test groups: 10 males and 10 females guinea pigs
- Control group: 10 males and 10 females guinea pigs
- Site: skin of the left flank
- Concentrations: 0.1 % solution of test substance in physiological saline
- Evaluation (hr after challenge): Twenty-four hours after the challenge injection

EPIDERMAL
- No. of exposures: one
- Day(s) of challenge: ten days after the challenge injection
- Exposure period: 24 h
- Test groups: 10 males and 10 females guinea pigs
- Control group: 10 males and 10 females guinea pigs
- Site: skin of the left flank
- Concentrations: 3 % of test substance in vaseline
- Evaluation (hr after challenge): Twenty-four hours after patch removal

OTHER:
Bodyweights were recorded immediately before starting the experiment (control values) and at termination of the study.

Positive control substance(s):

not specified

Results and discussion

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

0.1 ml of 0.1 % test item in physiological saline

No. with + reactions:

20

Total no. in group:

20

Remarks on result:

other: intradermal challenge

Results 2

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

30% of test item in vaseline

No. with + reactions:

0

Total no. in group:

20

Remarks on result:

no indication of skin sensitisation

Remarks:

epidermal challenge

Results 3

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

vehicle

No. with + reactions:

0

Total no. in group:

20

Any other information on results incl. tables

No difference between the test and the control group was seen after epidermal challenge application.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The negative results upon epidermal challenge demonstrate that, in artificially sensitised guinea-pigs, exposure of the intact skin to the test compound does not provoke contact dermatitis. The substance is not considered to be a skin sensitiser.

Executive summary:

Under the experimental conditions employed, significant differences between the test group and the vehicle treated controls were only seen after intradermal challenge application of test substance, i.e. when the skin barrier was intentionally by-passed. No difference between the test and the control group was seen after epidermal challenge application. The negative results upon epidermal challenge demonstrate that, in artificially sensitized guinea-pigs, exposure of the intact skin to the test compound does not provoke contact dermatitis.