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Toxicological information

Skin irritation / corrosion

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Administrative data

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
11-MAY-2004 to 19-AUG-2004
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
data is from experimental reports

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2004
Report Date:
2004

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Qualifier:
according to
Guideline:
other: Directive 92/69 EEC, B.4. "Acute Toxicity - Skin Irritation", July 31, 1992.
Principles of method if other than guideline:
The purpose of this primary skin irritation study was to assess the possible irritation potential when a single dose of C 010 was placed on the skin of rabbits for approximately four hours.
The test item was administered at 0.5 g/animal, the dose specified in the test guidelines for a solid test item.
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Details on test material:
- Name of test material (as cited in study report): Basic yellow 57
- Molecular formula (if other than submission substance): C19-H22-N5-O.Cl
- Molecular weight (if other than submission substance): 371.87 g/mol
- Smiles notation (if other than submission substance): N1(C([C@@H](\N=N\c2cc([N+](C)(C)C)ccc2)C(=N1)C)=O)c1ccccc1.[ClH-]
- InChl (if other than submission substance): 1S/C19H22N5O.ClH/c1-14-18(19(25)23(22-14)16-10-6-5-7-11-16)21-20-15-9-8-12-17(13-15)24(2,3)4;/h5-13,18H,1-4H3;1H/q+1;/p-1/b21-20+;
- Structural formula attached as image file (if other than submission substance): No data available
- Substance type: Organic
- Physical state: Solid orange powder
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Identification: C 010 (3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-N,N,N-trimethylanilinium chloride)
- Lot/batch No.of test material: 15
- Expiration date of the lot/batch: 30-DEC-2023
- Purity: 99.3% HPLC

RADIOLABELLING INFORMATION (Not applicable)
- Radiochemical purity:N/A
- Specific activity:N/A
- Locations of the label:N/A
- Expiration date of radiochemical substance:N/A

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: In the refrigerator (range of 5 ± 3°C), light protected.
- Stability under test conditions: Stable under storage conditions.
- Solubility and stability of the test substance in the solvent/vehicle: No data
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium:No data

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: 0.5 g (per animal) of C 010 was weighed as delivered by the Sponsor and then moistened with approximately 0.1 mL of purified water before application.
- Preliminary purification step (if any): No data
- Final dilution of a dissolved solid, stock liquid or gel: No data
- Final preparation of a solid: No data

FORM AS APPLIED IN THE TEST (if different from that of starting material) : No data

OTHER SPECIFICS:
Safety precautions: Routine hygienic procedures were used to ensure the health and safety of the personnel.

TEST ITEM PREPARATION
The pH of the test item was measured before the study initiation date. A formulation of a 1 % (w/w) solution was prepared. The pH was found to be 6.53.

Test animals

Species:
rabbit
Strain:
New Zealand White
Remarks:
SPF
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Elevage Scientifique des Dombes F-01400 Chatillon sur Chalaronne / France
- Age at study initiation: 12 weeks (male), 12 weeks (females)
- Weight at study initiation: Given below
- Identification: By unique cage number and corresponding ear number.
- Housing: Individually in stainless steel cages equipped with feed hoppers and drinking water bowls. Wood blocks (RCC Ltd,Füllinsdorf) and haysticks 4642 (batch no. 80/03, Provimi Kliba AG) were provided for gnawing.
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch no.24/04) provided by Provimi Kliba AG, CH-4303 Kaiseraugst.
- Water (e.g. ad libitum): Community tap water from Füllinsdorf, ad libitum.
- Acclimation period: 5 days. Under laboratory conditions after health examination. Only animals without any visual signs of illness were used for the study.
- Allocation: Male No. 82 , Female Nos. 83 and 84

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17-23°C
- Humidity (%): 30-70 %
- Air changes (per hr): Approximately 10-15 air changes per hour.
- Photoperiod (hrs dark / hrs light): The animals were provided with an automatically controlled light cycie of 12 hours light and 12 hours dark. Music was played during the daytime light period.

IN-LIFE DATES: From: 11-MA Y -2004 To: 19-AUG-2004

Test system

Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
water
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g

VEHICLE
- Amount(s) applied (volume or weight with unit): 0.1 mL of purified water

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): No data
- Concentration (if solution):No data

POSITIVE CONTROL
- Amount(s) applied (volume or weight): No data
- Concentration (if solution): No data
Duration of treatment / exposure:
4 hours
Observation period:
1, 24, 48 and 72 hours, as well as 7 and 10 days.
Number of animals:
3 (Animals of both sexes were used)
Details on study design:
TEST SITE
- Area of exposure: 100 cm2 (10 cm x 10 cm).
- % coverage: No data
- Type of wrap if used: The patch was covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and anchored with tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The skin was flushed with lukewarm tap water to clean the application site.
- Time after start of exposure: 4 hours

OBSERVATION TIME POINTS
(indicate if minutes, hours or days) : 1, 24, 48 and 72 hours, as well as 7 and 10 days.

SCORING SYSTEM:
- Method of calculation: The skin reaction was assessed according to the numerical scoring system listed in the EEC Commission Directive 92169/EEC, July 31, 1992 .

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
erythema score
Basis:
animal: 82,83,84
Time point:
24/48/72 h
Score:
0
Max. score:
3
Reversibility:
not specified
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
animal: 82,83,84
Time point:
24/48/72 h
Score:
0
Max. score:
3
Reversibility:
not specified
Remarks on result:
no indication of irritation
Irritation parameter:
erythema score
Basis:
animal: 82,83,84
Time point:
7 d
Score:
0
Max. score:
3
Reversibility:
not specified
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
animal: 82,83,84
Time point:
7 d
Score:
0
Max. score:
3
Reversibility:
not specified
Remarks on result:
no indication of irritation
Irritation parameter:
erythema score
Basis:
animal: 82,83,84
Time point:
10 d
Score:
0
Max. score:
3
Reversibility:
not specified
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
animal: 82,83,84
Time point:
10 d
Score:
0
Max. score:
3
Reversibility:
not specified
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
The animals were not sacrificed. No signs of irritation were observed on the treated skin following the application of the test item.
The mean score was calculated across 3 scoring times (24, 48 and 72 hours after patch removal) for each animal for erythemaJeschar grades and for oedema grades, separately.
The mean erythema/eschar score and the mean oedema score was 0.00 for all three animals.
No erythema and no swelling (oedema) was noted in any animal at any time.

CORROSION
Neither alterations of the treated skin were observed nor were corrosive effects evident on the skin.
Other effects:
- Other adverse local effects:
COLORATION
Slight yellow staining produced by the test item of the treated skin was observed in two animals at the 1-and 24-hour reading and persisted in one animal up to 7-days after
treatment.

BODY WEIGHTS
The body weights of all rabbits were considered to be within the normal range of variability.
Body weight in grams
Animal No. Sex First Day of Acclimatization Day of Treatment Last Day of Observation
82 male 2011 2109 2416
83 female 2113 2357 2754
84 female 2094 2247 2533


- Other adverse systemic effects:
VIABILITY/MORTALITY/CLINICAL SIGNS
No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.

Applicant's summary and conclusion

Interpretation of results:
other: Not irritating
Conclusions:
Based on these results and according to the CLP classification criteria: the test item C 010 (3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-N,N,N-trimethylanilinium chloride) is considered to be "not irritating" to rabbit skin.
Executive summary:

The primary skin irritation potential of C 010 (3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-N,N,N-trimethylanilinium chloride) was investigat ed according to OECD test guideline no. 404. The test item was applied by topical semi-occlusive application of 0.5 g to the intact left flank of each of three young adult New Zealand White rabbits. The duration of treatment was four hours. The scoring of skin reactions was performed 1, 24, 48 and 72 hours, as weil as 7 and 10 days after removal of the dressing.

The mean score was calculated across 3 scoring times (24, 48 and 72 hours after patch removal) for each animal for erythema/eschar grades and for oedema grades, separately.

The mean erythema/eschar score and the mean oedema score was 0.00 for all three animals.

The application of C 010 (3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-N,N,N-trimethylanilinium chloride) to the skin resulted in no signs of irritation. However slight yellow staining of the treated skin was observed in two animals at the 1- and 24-hour reading and persisted in one animal up to the 7-day examination. No corrosive effects were noted on the treated skin of any animal at any of the measuring intervals and no clinical signs were observed.

Thus, the test item did not induce significant or irreversible damage to the skin.

Based on these results and according to the CLP classification criteria: the test item C 010 (3-[(4,5-dihydro-3-methyl-5-oxo-1 -phenyl-1H- pyrazol- 4-yl)azo] -N,N, N-trimethylanilinium chloride) is considered to be "not irritating" to rabbit skin.