Chromate(2-), [3-[4,5-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-3-methyl-5-oxo-1H-pyrazol-1-yl]benzenesulfonato(3-)][2,4-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-5-methyl-2-phenyl-3H-pyrazol-3-onato(2-)]-, sodium

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP guideline study performed on a similar substance

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1995

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

other in vivo test already available

Test material

Test material information

Constituent 1

Reference substance name:

Chromate(2-), [2,4-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-5-methyl-2-phenyl-3H-pyrazol-3-onato(2-)][3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-2-hydroxy-5-nitrobenzenesulfonato(3-)]-, disodium

EC Number:

260-394-7

EC Name:

Chromate(2-), [2,4-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-5-methyl-2-phenyl-3H-pyrazol-3-onato(2-)][3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-2-hydroxy-5-nitrobenzenesulfonato(3-)]-, disodium

Cas Number:

56819-40-0

Molecular formula:

C32H21CrN10O11S.2Na

IUPAC Name:

Chromate(2-), [2,4-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-5-methyl-2-phenyl-3H-pyrazol-3-onato(2-)][3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-2-hydroxy-5-nitrobenzenesulfonato(3-)]-, disodium

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

None

In vivo test system

Test animals

Species:

guinea pig

Strain:

Hartley

Sex:

male/female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source:.From the results obtained under the experimental conditions employed, the test article did not provoke any reaction of cutaneous sensitization in the animals examined.
- Age at study initiation:about 6 weeks old
- Weight at study initiation:250 - 550 g.
- Housing:animals housed in groups according to EEC/86/609 in stainless steel mesh cages (internal dimensions 500 x 600 x 200 mm).
- Diet (e.g. ad libitum):ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period:at least 5 days between animal arrival and start of treatment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):19 3°C (target values)
- Humidity (%): >=45 % R.H. (target values),
- Air changes (per hr):>=22 air changes per hour,
- Photoperiod (hrs dark / hrs light):12 hours light (artificial)/12 hours dark.

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal

Vehicle:

water

Induction 2

Concentration / amount:

Treated group :
-By intradermal route : 3 series of 2 x 0.1 ml injections
*Freund's complete adjuvant at 50 % (V/V) in an isotonic injectable solution
*test article in a 0.1 % (W/W) solution in water for injection ;
*mixture 50/50 (V/V) : test article in a 0.2 % (W/W) solution in water for injection + Freund's complete adjuvant at 50 % (V/V) in an isotonic injectable solution, i.e. a final 0.1 % concentration of the test article.
During the preliminary study, injection of the test article in a 0.1 % solution tinted the skin of the animals thus making observation of erythema impossible. No oedema was noted.
-By topical occlusive route for 48 hours, with 0.5 ml of the test article in a 56 % (W/W) paste in water for injection.
During the preliminary study, the test article tinted the skin of the animals thus making observation of erythema impossible. Nevertheless as no oedema was noted, a skin painting was performed during the main study on Day 8, with 0.5 ml of sodium lauryl sulphate at 10 % (W/W) in Codex paraffin to create irritation.

.During the challenge, the topical occlusive application for 24 hours was performed in the treated group and in the control group with the test article in a 1 % (WIW) solution in water for injection and at the dose level of 0.5 ml (Maximum Non-Irritant Concentration : M.N.I.C.).

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

water

Challenge 2

Concentration / amount:

Treated group :
-By intradermal route : 3 series of 2 x 0.1 ml injections
*Freund's complete adjuvant at 50 % (V/V) in an isotonic injectable solution
*test article in a 0.1 % (W/W) solution in water for injection ;
*mixture 50/50 (V/V) : test article in a 0.2 % (W/W) solution in water for injection + Freund's complete adjuvant at 50 % (V/V) in an isotonic injectable solution, i.e. a final 0.1 % concentration of the test article.
During the preliminary study, injection of the test article in a 0.1 % solution tinted the skin of the animals thus making observation of erythema impossible. No oedema was noted.
-By topical occlusive route for 48 hours, with 0.5 ml of the test article in a 56 % (W/W) paste in water for injection.
During the preliminary study, the test article tinted the skin of the animals thus making observation of erythema impossible. Nevertheless as no oedema was noted, a skin painting was performed during the main study on Day 8, with 0.5 ml of sodium lauryl sulphate at 10 % (W/W) in Codex paraffin to create irritation.

.During the challenge, the topical occlusive application for 24 hours was performed in the treated group and in the control group with the test article in a 1 % (WIW) solution in water for injection and at the dose level of 0.5 ml (Maximum Non-Irritant Concentration : M.N.I.C.).

No. of animals per dose:

30 animals of both sexes, allocated to one control group of 10 animals (induction: vehicle - challenge : test article) and one treated group of 20 animals (induction and challenge: test article).

Details on study design:

.During induction, the applications were performed as follows:
. Treated group:
-By intradermal route: 3 series of 2 x 0.1 ml injections
*Freund's complete adjuvant at 50 % (V/V) in an isotonic injectable solution
*test article in a 0.1 % (W/W) solution in water for injection ;
*mixture 50/50 (V/V): test article in a 0.2 % (W/W) solution in water for injection + Freund's complete adjuvant at 50 % (V/V) in an isotonic injectable solution, i.e. a final 0.1 % concentration of the test article.
During the preliminary study, injection of the test article in a 0.1 % solution tinted the skin of the animals thus making observation of erythema impossible. No oedema was noted.
-By topical occlusive route for 48 hours, with 0.5 ml of the test article in a 56 % (W/W) paste in water for injection.
During the preliminary study, the test article tinted the skin of the animals thus making observation of erythema impossible. Nevertheless as no oedema was noted, a skin painting was performed during the main study on Day 8, with 0.5 ml of sodium lauryl sulphate at 10 % (W/W) in Codex paraffin to create irritation.

Control group:
The intradermal injections and the topical occlusive application for 48 hours were carried out under the same conditions as in the treated group, water for injection replacing the test article.
The rest period was 11 days without treatment.
During the challenge, the topical occlusive application for 24 hours was performed in the treated group and in the control group with the test article in a 1 % (WIW) solution in water for injection and at the dose level of 0.5 ml (Maximum Non-Irritant Concentration: M.N.I.C.).
The cutaneous macroscopic examinations were performed 24 and 48 hours after removal of the patches to the challenge application site, according to the Magnusson & Kligman scale. As the test article tinted the skin of the animals, thus making observation of erythema impossible, histopathological examinations of the skin were performed for all the animals of the treated and control groups (in half of them at 24 hours and in the other half at 48 hours).

Challenge controls:

one control group of 10 animals (induction : vehicle - challenge : test article)

Positive control substance(s):

not specified

Results and discussion

Positive control results:

None

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

48

Group:

test chemical

Dose level:

1%

No. with + reactions:

0

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 1%. No with. + reactions: 0.0.

Results 2

Reading:

rechallenge

Hours after challenge:

48

Group:

test chemical

Dose level:

1%

No. with + reactions:

0

Remarks on result:

other: Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 1%. No with. + reactions: 0.0.

Any other information on results incl. tables

Determination of a weak to moderate irritant concentration by intradermal injection

Evaluation of the reactions at different observation times
		24 hours			48 hours
After the intradermal injections
Sex/Guinea pig No	Concentrations	1%	0.10%	0.05%	1%	0.10%	0.05%
M/61675	Erthema    (+Oedema)	3	?	?	?	?	?
	Other anomaly	Y	U	U	U	U	U
M/61676	Erthema    (+Oedema)	?	?	?	?	?	?
	Other anomaly	U	U	U	U	U	U
F/61677	Erthema    (+Oedema)	3	?	?	?	?	?
	Other anomaly	Y	U	U	U	U	U
F/61678	Erthema    (+Oedema)	?	?	?	?	?	?
	Other anomaly	U	U	U	U	U	U

? - Reading impossible

U - The test article tinted the skin of the animals thus making erythema observation impossible. No oedema was noted.

Y - The test article tinted the skin making erythema observation impossible. The maximum score (score = 3) was attributed because of the presence of oedema.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Remarks:

Migrated information

Conclusions:

From the results obtained under the experimental conditions employed, the test article did not provoke any reaction of cutaneous sensitization in the animals examined.

Executive summary:

A GLP compliant OECD guideline 406 test was performed in the albino Hartley guinea-pig (supplier Charles River France), to evaluation of the delayed cutaneous hypersensitivity of the test article according to a maximized protocol using 30 animals of both sexes, allocated to one control group of 10 animals (induction : vehicle - challenge : test article) and one treated group of 20 animals (induction and challenge : test article).

Animals were exposed to test article by 3 series of 2 X 0.1 ml injections. Test solution was made as 0.1% (W/W) solution in water for injection.

During the preliminary study, the test article tinted the skin of the animals thus making observations of erythema impossible. Nevertheless as no oedema was noted, a skin painting was performed during the main study on Day 8, with 0.5 ml of sodium lauryl sulphate at 10% (W/W) in Codex paraffin to create irritation.

After a rest period of 11 days, challenge study was performed with the topical occlusive application for 24 h in treated and control group with the test article in a 1% (W/W) solution in water for injection and at the dose level of 0.5 ml (Maximum Non-irritant concentration)

The cutaneous macroscopic examination were performed 24 and 48 h after removal f the patches to the challenge application site, according to the Magnusson & Kligman scale. As the test article tinted the skin of the animals, thus making observations of erythema impossible, histopathological examination of the skin were performed for all the animals of the treated and control groups.

Signs of irritation were noted during induction after application of sodium lauryl sulphate in both groups. After challenge, the macroscopic and histopathological examinations did not reveal any lesion of delayed hypersensitivity in the 20 treated animals. No noticeable cutaneous abnormality was noted in the 10 guinea-pigs examined in the control group.

From the results obtained under the experimental conditions employed, the test article did not provoke any reaction of cutaneous sensitization in the animals examined. Hence, the test substance is regarded as non-sensitizer.