Registration Dossier

Administrative data

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
23 - 27 Feb 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP - Guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2011
Report Date:
2011

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2500 (Acute Dermal Irritation)
Qualifier:
according to
Guideline:
other: JMAFF 12-Nousan-8147 (2000)
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
Animals: New Zealand White rabbits
Source: Inhouse Bred (Outbred) New Zealand White rabbits Toxicology Department of Safety Assessment, Advinus Therapeutics Limited, Bangalore 560 058, INDIA
Number and sex: 3 Male rabbits
Age at treatment: 6-7 months
Date of birth: 30 July 2010
Body weight range at treatment: 2.77 to 2.94 kg
Accl imatization: One rabbit (RB8764) was acclimatized for five days while the other two (RB8765 and RB8766) rabbits were accl imatized for six days under laboratory conditions after physical examination.

Conditions
Animals were housed under standard laboratory conditions, air conditioned with adequate fresh air supply (12-15 air changes/hour). Environment: temperature 21 - 23°C, relative humidity 65 - 67%, with a 12 hour light and 12 hour dark cycle. The maximum and minimum temperature and relative humidity in the experimental room were recorded once daily. The relative humidity in the experimental room was calculated from dry and wet bulb temperature recordings.

Housing
The rabbits were housed in dividually in rabbit cages (approx. size: L 65 x B 65 x H 45 cm) with Noryl shallow cage body and facilities for pelleted food (Stainless steel feed hopper) and drinking water (750 mL markrolon bottle fitted with sipper tube). The litter collection tray (noryl waste tray) were washed daily with water (except on Sundays). Water was replenished daily. The water bottles and feed hoppers were changed once a week.

Diet: ad libitum
The animals were offered rabbit feed manufactured by Pranav agro Industries Ltd, Sangli.

Specific contaminants (heavy metals organochlorines and organophosphates) in the diet are analysed periodically and the reports are included in the raw data file.

Water: ad libitum
The animals were provided deep bore-well water passed through activated charcoal filter and exposed to UV rays in Aquaguard on-line water filter-cumpurifier manufactured by Eureka Forbes Ltd. , Mumbai - 400 00 I, India. The water bottles were replenished once daiIy and the water bottles changed once a week.

Test system

Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
water
Controls:
other: each rabbit also served as it's own control with untreated areas of the skin.
Amount / concentration applied:
Sample preparation
A quantity of 0.5 g of test substance as received was made into paste by adding sufficient volume of water and mixed with a glass rod.

Preparation of Test Animals
Approximately 24 hours prior to treatment, the fur on the left side of the dorsolateral trunk surface [about 14 x 19 cm] was clipped with an electric clipper -Aesculap® - Germany. Care was taken to avoid abrading the skin. After clipping and prior to application, the animal 's skin (test site) was checked for any abnormalities according to the "Primary Skin Irritation Scoring System", and the test site was found to be normal.

Duration of treatment / exposure:
4 hours
Observation period:
3 days: skin reactions were evaluated 1 hour and then 24, 48 and 72 hours after removal of the patch
Number of animals:
3
Details on study design:
Testing Option
The study was initiated in a stepwise procedure (1 + 2 Rabbits).

Test substance Application
Preparation of Dose Site
Approximately 6 cm2 intact test site was selected on each animal. The adjacent untreated skin of each animal served as the control site.

Application of Test Substance
A quantity of 0.5 g of test substance as received was made into paste by adding sufficient volume of water and mixed with a glass rod. The test substance was applied directly to the skin of the test site and covered with a gauze pad. The entire trunk of each animal was then wrapped with non-irritating semiocclusive adhesive tape to avoid dislocation of the patch.

Exposure of Animals
In a stepwise manner (1 +2 rabbits) all the three rabbits were exposed to the test substance each with one test patch only for a four hour exposure period. The rabbits were restrained using Elizabethan collar for 72 hours post application of the test patch.

Toxic Signs and Pre-Terminal Deaths
Rabbits were observed for toxic signs and pre-terminal deaths for the first four hours (at hourly intervals) on the day of test substance application and once daily thereafter till the end of the observation period.

Body Weights
Individual body weights of the rabbits were recorded at the start of acclimatization, day 1 of treatment (just before test substance application) and
at the termination of observation.

Skin Scoring
Following the exposure period, test patches were removed and the application sites were washed with water and clean towels to remove any residual test substance. The 4 hour dose sites were examined for signs of erythema and edema within 60 minutes and at intervals of approximately 24, 48 and 72 hours after test patch removal accord ing to the "Primary Skin Irritation Scoring System" developed by Draize.

Individual skin scores were recorded for each rabbit.

Grading of skin reactions
The skin reaction was assessed according to the numerical scoring system of Draize et.al (1944).
1. ERYTHEMA AND ESCHAR FORMATION
No erythema.................... . ...................................................................................................................................................................... 0
Very slight erythema (barely perceptible)........................................................................................................................................... 1
Well-defined erythema........................................................................................................................................................................... 2
Moderate to severe erythema.......... .................................................................................................................................................... 3
Severe erythema (beet redness) to slight eschar formation (injuries in depth) .......................................................................... 4
MAXIMUM POSSIBLE SCORE - 4

2. EDEMA FORMATION
No edema.................................................................................................................................................................................................. 0
Very sl ight edema (barely perceptible) .............................................................................................................................................. 1
Slight edema (edges of area well defined by definite raising) ......................................................................................................... 2
Moderate edema (raised approximately 1 millimetre).... .................................................................................................................. 3
Severe edema (raised more than 1 millimetre and extending beyond area of exposure)........................................................... 4
MAXIMUM POSSIBLE SCORE - 4

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
erythema score
Basis:
mean
Remarks:
out of all 3 animals
Time point:
other: 24, 48, 72 hours
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Irritation parameter:
edema score
Basis:
mean
Remarks:
out of all 3 animals
Time point:
other: 24, 48, 72 hours
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Irritant / corrosive response data:
There were no skin reactions observed In any of the rabbits through the observation period.
Other effects:
There were no toxic signs and pre-terminal deaths during the course of the experiment. The body weights of all the rabbits increased through the observation period.

Any other information on results incl. tables

Erythema and edema scores at 1, 24, 48 and 72 hours were 0 for each of the three animals tested.

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
CLP: not classified
DSD: not classified