Registration Dossier

Toxicological information

Skin irritation / corrosion

Currently viewing:

Administrative data

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Principles of method if other than guideline:
The objective of the study was to assess the irritant and/or corrosive effects of Solvent Blue 4 (CAS No. - 6786-83-0) after dermal application on the intact skin in Rabbits
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report):Solvent Blue 4
- Molecular formula :C33H33N3O
- Substance type:Organic
- Storage condition of test material:Store in a cool, dry place, tightly sealed containers protected from light. Keep away from possible sources of ignition.
- Other:
Handling and Disposal
Safety precautions : Avoid ingestion and inhalation of the neat material. Aprons,caps, mask, gloves and goggles were used to ensure the health and safety of the Personnel.
Disposal : The remaining unused test item was disposed as per internal SOPs of sa-FORD.

Test animals

Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:Procured from LIVEON BIOLABS PVT LTD, Karnataka, India.
- Age at study initiation:4.5 to 5.0 Months (Approximately)
- Weight (Prior to Treatment):Minimum: 2.038 kg & Maximum: 2.548 kg (Prior to Treatment)
- Health Status : Healthy young adult rabbits were used for the study
-Diet:All animals were provided conventional laboratory rabbit diet (Nutrivet Life Sciences, Pune) ad libitum. Batch No.: 200004.
-Water:Aqua guard filtered tap water was provided ad libitum.
-Husbandry: The animals were housed individually in stainless steel cages.
-Room Sanitation:The experimental room floor and work tops were swept and mopped with disinfectant solution every day.
-Cages and water bottle:All the cages and water bottles were changed minimum twice a week
-Acclimatisation:Rabbits were acclimatised to the test conditions for a period of 6 days (Animal No.-1) and 8 days (Animal No.-2 and 3) prior to the application of the test item

ENVIRONMENTAL CONDITIONS
Temperature: Minimum: 20.30 °C Maximum: 22.70 °C
Relative humidity: Minimum: 48.30 % Maximum: 68.30 %
Light-dark-rhythm: 12:12
Air Changes:More than 12 changes per hour


Test system

Type of coverage:
other: porous gauze dressing and non-irritating tape
Preparation of test site:
clipped
Vehicle:
other: distilled water
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit):0.5 ml
- Concentration (if solution):N/A

VEHICLE
- Amount(s) applied (volume or weight with unit):500 mg of test item pulverised form (moistened with 0.5 ml distilled water
- Concentration (if solution):N/A
- Lot/batch no. (if required):N/A
- Purity:N/A
Duration of treatment / exposure:
4 hours
Observation period:
72 hours
Number of animals:
3 male rabbits
Details on study design:
TEST SITE
- Area of exposure:The dorsal lumbar region at contralateral sites.
- % coverage:Approximately 6 X 6 cm.
- Type of wrap if used:A porous gauze dressing and non-irritating tape (Micropore 3”).

REMOVAL OF TEST SUBSTANCE
- Washing (if done):The residual test item was removed by using cotton soaked in distilled water.
- Time after start of exposure:4 hour

SCORING SYSTEM:Draize Method

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
other: At 24 hours
Score:
0.33
Max. score:
3
Reversibility:
no data
Remarks on result:
other: not irritating
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
other: 48hr
Score:
0.67
Max. score:
3
Reversibility:
no data
Remarks on result:
other: not irritating
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
other: 72hr
Score:
0.67
Max. score:
3
Reversibility:
no data
Remarks on result:
other: not irritating
Irritation parameter:
edema score
Basis:
animal #1
Time point:
other: At 24, hours
Score:
0
Max. score:
3
Reversibility:
no data
Remarks on result:
other: not irritating
Irritation parameter:
edema score
Basis:
animal #2
Time point:
other: 48hrs
Score:
0
Max. score:
3
Reversibility:
no data
Remarks on result:
other: not irritating
Irritation parameter:
edema score
Basis:
animal #3
Time point:
other: 72hrs
Score:
0.33
Max. score:
3
Reversibility:
no data
Remarks on result:
other: not irritating
Irritant / corrosive response data:
The following were observed in treated rabbits.
The patch was removed after 4 hours and rabbits were observed for erythema and oedema at 1, 24, 48 and 72 hours after patch removal, evaluated and graded as per draize method.
Animal No. 1, showed very slight erythema (barely perceptible) and very slight oedema (barely perceptible) at 1 hour of observation. At 24 hour very slight erythema (barely perceptible) and no oedema was observed. At 48 and 72 hour observation no erythema and oedema was observed in animal no 1.
In Animals No. 2 and 3 at 1 hour observation post patch removal, revealed very slight erythema (barely perceptible) and very slight oedema (barely perceptible). At 24 hour observation post patch removal, revealed very slight erythema (barely perceptible) and no oedema in animal no. 2 whereas very slight erythema (barely perceptible) and very slight oedema (barely perceptible) was observed in animal no. 3. At 48 hour post patch removal very slight erythema (barely perceptible) and no oedema was observed. At 72 hour post patch removal, both the animals recovered to normal.
The individual mean score at 24, 48 and 72 hours for Animal Nos. 1, 2 and 3 were 0.33, 0.67, 0.67 and 0.00, 0.00, 0.33, for erythema and oedema formation, respectively..
Other effects:
Clinical Observation
No systemic toxicity was observed at treated rabbits during the experimental period.

Mortality
No mortality was observed during the observation period.

Body weight
All rabbits showed increased in body weight at termination when compared to day 0

Any other information on results incl. tables

Table 1

Skin Reaction

In Treated area Dose:500 mg of test item (moistened with 0.5 ml distilled water)           Sex:Male

 

Animal

No.

Test

Treated

 area*

Erythema score

Oedema score

1h

24h

48h

72h

1h

24h

48h

72h

1

Initial

Left

1

1

0

0

1

0

0

0

2

Confirmatory

Left

1

1

1

0

1

0

0

0

3

Left

1

1

1

0

1

1

0

0

 

 

 

 

 

In Control areaDose:0.5 ml of distilled water                                                          Sex:Male

 

Animal

No.

Test

Treated area*

Erythema score

Oedema score

1h

24h

48h

72h

1h

24h

48h

72h

1

Initial

Right

0

0

0

0

0

0

0

0

2

Confirmatory

Right

0

0

0

0

0

0

0

0

3

Right

0

0

0

0

0

0

0

0

Key: h = Hour.

Erythema                                                                               Oedema

0 =No erythema                                                                   0 =No oedema

1 = Very slight erythema(barely perceptible)                       1 =very slight oedema (barely perceptible)

Table 1 Continued…

Mean Individual Animal Score at 24, 48 and 72 hours

 

                     Animal Number                  

Observations                      

1

2

3

Erythema

0.33

0.67

0.67

Oedema

0.00

0.00

0.33

 

  

Table 2

Individual Animal BodyWeight

Sex:Male

Animal

No.

Body Weight (kg)

Prior to Dosing

At termination

1

2.412

2.466

2

2.548

2.614

3

2.038

2.104

 

Table 3

Individual AnimalClinicalSigns

Sex:Male

Animal

No.

Days (Post dosing Observation)

0

1

2

3

1

1

1

1

1

2

1

1

1

1

3

1

1

1

1

Key: ./. = Not Applicable. 1 = Normal

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The individual mean score at 24, 48 and 72 hours for animal nos. 1, 2 and 3 were 0.33, 0.67, 0.67 and 0.00, 0.00, 0.33, for erythema and oedema formation, respectively. Hence, it was concluded that Solvent Blue 4 (CAS No. 6786-83-0) was Non-Irritating to the skin of male New Zealand White rabbits under the experimental conditions as per CLP Regulation
.

Executive summary:

Acute Dermal Irritation/corrosion Study of Solvent Blue 4 (CAS No:6786-83-0)” was carried out in Rabbits, This study was performed as per OECD guideline No. 404.

Three healthy young adult male rabbits were used for conducting acute dermal irritation study.Body weights were recorded on day 0 (prior to application) and at termination.

Rabbits with good intact skin were selected for the study. The hairs of all the rabbits were clipped at contralateral sites, approximately 24 hours prior to treatment.A dose of 0.5 ml of test item (as such) was applied to the skin,over an area of approximately 6 x 6 cm clippedof hair on one side of rabbits.The other untreated side was kept as control area and 0.5 ml of distilled water was applied at this site. At the end of 4 hours, the gauze patch was removed and test item application site was wiped with water without altering the integrity of the epidermis. 

Initially, the test item was applied to the clipped area of skin of one rabbit. The test site was covered with gauze patch.After 4 hours of exposure in animal no. 1, there was very slight erythema (barely perceptible) and very slight oedema (barely perceptible) observed at 1 hour of observation. At 24 hour very slight erythema (barely perceptible) and no oedema was observed.Hence the confirmatory test was conducted on additional two animal (No. 2 and 3)to confirm the non irritant nature of the test item. At 48 and 72 hours observation no erythema and no oedema was observed in animal no 1.

The patch was removed after 4 hours and rabbits wereobservedfor erythema and oedemaat 1, 24, 48 and 72 hours after patch removal, evaluatedand graded as per draize method.In animals no. 2 and 3 at 1 hour observation post patch removal, revealedvery slight erythema (barely perceptible) and very slight oedema (barely perceptible). At 24 hour observation post patch removal, revealed very slight erythema (barely perceptible) and no oedema in aniaml no. 2 whereasvery slight erythema (barely perceptible) and very slight oedema (barely perceptible), in animal no. 3. At 48 hour post patch removal very slight erythema (barely perceptible) and no oedema was observed in animal no. 2 and 3. At 72 hour post patch removal, both the animals recovered to normal.The individual mean score at24, 48 and 72 hoursfor animal nos. 1, 2 and 3 were 0.33, 0.67, 0.67 and 0.00, 0.00, 0.33, for erythema and oedema formation, respectively.  

Hence, it was concluded that Solvent Blue 4 (CAS No. - 6786-83-0) was Non-Irritating to the skin of female New Zealand White rabbits under the experimental conditions tested and not Classified as a "skin irritant" as per CLP Regulation.