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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
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:
2014
Report Date:
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 Reaction mass of test chemical after dermal application on the intact skin in Rabbits.
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
- Name of test material: Reaction mass of 4-(2,6,6-trimethylcyclohex-2-ene-1-yl)-but-3-ene-2-one and 4-(2,6,6-trimethylcyclohex-1-ene-1-yl)-but-3-ene-2-one
- IUPAC name: Reaction mass of 4-(2,6,6-trimethylcyclohex-2-ene-1-yl)-but-3-ene-2-one and 4-(2,6,6-trimethylcyclohex-1-ene-1-yl)-but-3-ene-2-one
- Molecular formula: C13H20O
- Molecular weight: 192.3 g/mole
- Smiles : C1([C@@H](C(=CCC1)C)\C=C\C(C)=O)(C)C
- Inchl: 1S/C13H20O/c1-10-6-5-9-13(3,4)12(10)8-7-11(2)14/h6-8,12H,5,9H2,1-4H3/b8-7+
- Substance type: Organic
- Physical state: Solid (yellow)

Test animals

Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Sex: Female
- Source: Procured from RABBI ROOF, Hyderabad, 500075, (CPCSEA Reg. No. 1655/PO/b/12/CPCSEA).
- Age at study initiation: 3.5 to 4.5 Months (Approximately)
- Health Status: Healthy young adults and nulliparous and non-pregnant Rabbits were used for the study.
- Weight at study initiation: Minimum: 1.724 kg & Maximum: 1.926 kg (Prior to Treatment)
- Housing: The animals were housed individually in stainless steel cages.
- Diet (e.g. ad libitum): All animals were provided conventional laboratory rabbit diet (Nutrivet Life Sciences, Pune) ad libitum. Batch No.: 200012.
- Water (e.g. ad libitum): Aqua guard filtered tap water was provided ad libitum.
- Acclimation period: Rabbits were acclimatised to the test conditions for a period of 5 days (Animal No.-1) and 9 days (Animal No.-2 and 3) prior to the application of the test item
- Identification:During acclimatization marking was done with non toxic marker pen in the inside of left ear of rabbits and after acclimatization, animals were marked with permanent number in the inner side of right ear of rabbits. Permanent marker and cage card was used for identification.
- 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.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Minimum: 19.30 °C and Maximum: 22.50 °C
- Humidity (%): Minimum: 47.00 % and Maximum: 68.40 %
- Air changes (per hr): More than 12 changes per hour
- Photoperiod (hrs dark / hrs light): 12:12

IN-LIFE DATES: From: To:

Test system

Type of coverage:
occlusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent vehicle
Amount / concentration applied:
0.5 ml
Duration of treatment / exposure:
4 hours
Observation period:
72 hours
Number of animals:
3 female
Details on study design:
TEST SITE
- Area of exposure:dorsal lumbar region
- % 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): yes
- Time after start of exposure:At the end of the 4 hour exposure period, residual test item was removed by using cotton soaked in distilled water.

OBSERVATION TIME POINTS
(indicate if minutes, hours or days) : patches were removed after 4 hours and the rabbits were observed for a maximum period of 72 hours for grading

SCORING SYSTEM:
- Method of calculation:Grading of irritation lesions was carried out as per Draize Method.

Results and discussion

In vitro

Other effects / acceptance of results:
no data available

In vivo

Resultsopen allclose all
Irritation parameter:
erythema score
Basis:
animal: 1,2,3
Time point:
72 h
Score:
0
Reversibility:
not specified
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
animal: 1,2,3
Time point:
72 h
Score:
0
Reversibility:
not specified
Remarks on result:
no indication of irritation
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 no oedema observed at 1 hour of observation. At 24 hour, 48 hour and 72 hour observation no erythema and oedema was observed in animal No 1.
In Animals No. 2 and 3 at 1 hour observation after post patch removal, revealed no erythema and oedema. At 24 and 48 and 72 hour observation animals showed no erythema post patch removal. No oedema was observed during the observation period.
The individual mean score at 24, 48 and 72 hours for Animal Nos. 1, 2 and 3 were 0.00, 0.00, 0.00 and 0.00, 0.00, 0.00, 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.

Any other information on results incl. tables

Table 1: Skin Reaction

 

In Treated area Dose:0.5 ml of test item                                                               Sex:Female

 

Animal

No.

Test

Treated

 area*

Erythema score

Oedema score

1h

24h

48h

72h

1h

24h

48h

72h

1

Initial

Right

1

0

0

0

0

0

0

0

2

Confirmatory

Left

0

0

0

0

0

0

0

0

3

Right

0

0

0

0

0

0

0

0

  

In Control area Dose:0.5 ml of distilled water                                                Sex:Female

 

Animal

No.

Test

Treated area*

Erythema score

Oedema score

1h

24h

48h

72h

1h

24h

48h

72h

1

Initial

Left

0

0

0

0

0

0

0

0

2

Confirmatory

Right

0

0

0

0

0

0

0

0

3

Left

0

0

0

0

0

0

0

0

Key: h = Hour.

Erythema                                                                                                       Oedema

0 =No erythema                                                                                           0 =No oedema


Mean Individual Animal Score at 24, 48 and 72 hours

 

                     Animal Number                  

Observations                      

1

2

3

Erythema

0.00

0.00

0.00

Oedema

0.00

0.00

0.00

 

Table 2: Individual Animal BodyWeight

Sex:Female

Animal

No.

Body Weight (kg)

Prior to Dosing

At termination

1

1.926

2.050

2

1.724

1.808

3

1.858

1.920

 

Table 3: Individual AnimalClinicalSigns

Sex:Female

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:
other: not irritating
Conclusions:
No erythema and oedema (skin irritation) were found at the end of 72 hour observation period after patch removal. The individual mean score at 24, 48 and 72 hours for Animal Nos. 1, 2 and 3 were 0.00, 0.00, 0.00 and 0.00, 0.00, 0.00, for erythema and oedema formation, respectively.Hence, it was concluded that the test chemical was Non-Irritating to the skin of Female New Zealand White rabbits under the experimental conditions tested.
Executive summary:

Acute Dermal Irritation/corrosion Study of test chemical in Rabbits, was performed in an OECD GLP laboratory (Sustainability Support Services (Europe) AB has the letter of access) as per OECD guideline No. 404. Three healthy young adult female rabbits were used for conducting acute dermal irritation/corrosion 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 test item was applied to the skin, over an area of approximately 6 x 6 cm clipped of 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 No oedema observed at 1 hour of observation. At 24, 48 and 72 hours observation no erythema and oedema was observed in animal No 1.Hence the confirmatory test was conducted on additional two rabbits (No. 2 and 3) to confirm the non irritant nature of the test item. 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. In Animals No. 2 and 3 at 1 hour observation after post patch removal, revealed no erythema and oedema. At 24 and 48 and 72 hour observation animals showed no erythema post patch removal. No oedema was observed during the observation period. The individual mean score at 24, 48 and 72 hours for Animal Nos. 1, 2 and 3 were 0.00, 0.00, 0.00 and 0.00, 0.00, 0.00, for erythema and oedema formation, respectively. Hence, it was concluded that the test substance was Non-Irritating to the skin of Female New Zealand White rabbits under the experimental conditions tested.