Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Skin Irritation

A Preliminary irritation tests for test chemical were performed on male albino Dunkin/Hartley guinea pigs to determine the concentration for suitable the sensitization study. In the preliminary test, the guinea pigs were treated intradermally and dermally. Since the test chemical induced cutaneous effects at these concentrations, the test chemical was considered to be slightly irritating to the skin of albino Dunkin/Hartley guinea pigs. Thus concentration of 0.5% was selected for the intradermal induction and a concentration of 50% the test chemical was selected for the topical induction application. Based on the observations and results, it was concluded that the test material was considered to be slightly irritating (Category 2) on male albino Dunkin/Hartley guinea pigs in the preliminary irritation tests performed for the sensitization study.

Eye Irritation

Based on the available results and applying the weight of evidence approach the test chemical can be considered to be not irritating to the eyes and can be classified under the category “Not Classified” as per CLP Regulation.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
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 report
Qualifier:
according to guideline
Guideline:
other: as mentioned below
Principles of method if other than guideline:
Preliminary irritation tests for test chemical were performed in male Albino Dunkin/Hartley guinea pigs to determine the concentration for suitable the sensitization study.
GLP compliance:
not specified
Species:
guinea pig
Strain:
other: Albino Dunkin/Hartley
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: No Data Available
- Age at study initiation: No Data Available
- Weight at study initiation: 400-500 grams 320 g (intradermal exposure ) and 450g (dermal exposure )
- Housing: No Data Available
- Diet (e.g. ad libitum): RGP pellets, hay cabbage ad libitum
- Water (e.g. ad libitum): water ad libitum
- Acclimation period: No Data Available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): No Data Available
- Humidity (%): No Data Available
- Air changes (per hr): No Data Available
- Photoperiod (hrs dark / hrs light): No Data Available

IN-LIFE DATES: From: To: No Data Available
Type of coverage:
occlusive
Preparation of test site:
other: Intradermal application: clipped flanks Dermal application: shaved flanks
Vehicle:
other: Intradermal application: 6% acetone/20% PEG400/0.01% Tween 80/saline Dermal application: acetone
Controls:
not specified
Amount / concentration applied:
Intradermal concentration: 0.25 %,0.5%, 1% or 2%
Topical Concentration: 10%, 25 % or 50%
Duration of treatment / exposure:
Single application
Observation period:
24 and 48 hours
Number of animals:
4 (same sex)
Details on study design:
Area of exposure: shaved flanks
- % coverage: 8mm diameter filter paper patches in 11mm aluminum patch test cups are saturated with a range of concentrations of test chemical.
- Type of wrap if used: the patches are held in place by adhesive plaster ound around the trunk.
- Time after start of exposure: 24 and 48 hours after removal of the patches.
SCORING SYSTEM: the resulting reactions are scored for irritation on a scale from 0 to +++. (mentioned in table 1)
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
24/48 h
Reversibility:
not specified
Remarks on result:
other: Very slight erythema was observed at 0.25% and 0.5% . Barely perceptible erythema was observed at 10%, 25% or 50% concentrations
Irritant / corrosive response data:
Very slight erythema was observed at 0.25% and 0.5% . Barely perceptible erythema was observed at 10%, 25% or 50% concentrations
Other effects:
No Data Available

Table 1.Skin Reaction Scoring System

SKIN REACTIONS

SCORES

No reaction

0

Barely perceptible erythema

±

Scattered, mild erythema (faint pink)

+

Moderate and diffuse erythema (pale pink )

++

Intense erythema (deep pink) and oedema

+++

Table 2: RESULTS OF PRELIMINARY IRRITATION TEST

Guinea pigs

Concentrations

No.

Sex

10%

25%

50%

24 hr

48 hr

24 hr

48 hr

24 hr

48 hr

1

M

0

0

±

±

±

±

2

M

0

0

0

0

±

±

3

M

±

0

±

±

±

4

M

0

0

±

0

0

0

Interpretation of results:
Category 2 (irritant) based on GHS criteria
Conclusions:
Based on the observations and results, it was concluded that the test material was considered to be slightly irritating (Category 2) on male albino Dunkin/Hartley guinea pigs in the preliminary irritation tests performed for the sensitization study.
Executive summary:

A Preliminary irritation tests for test chemical were performed on male albino Dunkin/Hartley guinea pigs to determine the concentration for suitable the sensitization study. In the preliminary test, the guinea pigs were treated intradermally and dermally. The animals were intradermally injected with 0.1ml aliquots of a range of concentration of test substance (0.25%,0.5%, 1% or 2%) in 6% acetone/ 20% PEG 400 /0.01% Tween 80/ saline. During topical application, animals received a single dermal application of 10%, 25% or 50% test chemical in acetone using 8 mm saturated filter paper and 11 mm aluminum patch test cups which were applied to the shaved flank for 24 h. Reactions were assessed for irritation at 24 and 48 h after patch removal. Very slight erythema was observed at 0.25% and 0.5%. Barely perceptible erythema was observed at 10%, 25% or 50% concentrations. Since the test chemical induced cutaneous effects at these concentrations, the test chemical was considered to be slightly irritating to the skin of albino Dunkin/Hartley guinea pigs. Thus concentration of 0.5% was selected for the intradermal induction and a concentration of 50% the test chemical was selected for the topical induction application. Based on the observations and results, it was concluded that the test material was considered to be slightly irritating (Category 2) on male albino Dunkin/Hartley guinea pigs in the preliminary irritation tests performed for the sensitization study.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Referenceopen allclose all

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Remarks:
Read-across from supporting substance (surrogate)
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
data is from experimental reports
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Principles of method if other than guideline:
The objective of the study was to assess the irritant and/or corrosive effects of the test chemical on eye, when exposed by the ocular route in rabbits.
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Sex: Male
- Source: Procured from LIVEON BIOLABS PVT. LTD.
- Age at study initiation: 3.0 to 4.5 Months (Approximately)
- Weight at study initiation: Minimum: 1.822 kg and Maximum: 2.146 kg (Prior to Treatment)
- Health Status : Healthy young adult animals were used for the study.
- Housing: 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.
- Diet (e.g. ad libitum): All animals were provided conventional laboratory rabbit diet, ad libitum.
- 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 6 days (Animal No.-1) and 9 days (Animal No. 2 and 3) prior to the application of the test item.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):Minimum: 19.00 °C and Maximum: 20.80 °C
- Humidity (%):Minimum: 45.10 % and Maximum: 65.80 %
- Air changes (per hr): More than 12 changes per hour
- Photoperiod (hrs dark / hrs light): 12:12
Vehicle:
unchanged (no vehicle)
Controls:
yes
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit):0.1 ml
- Concentration (if solution):N/A

VEHICLE
- Amount(s) applied (volume or weight with unit):N/A
- Concentration (if solution):N/A
- Lot/batch no. (if required):N/A
- Purity:N/A
Duration of treatment / exposure:
24 hrs
Observation period (in vivo):
All the treated animals were observed at 1, 24, 48 and 72 hours after instillation of test item
Duration of post- treatment incubation (in vitro):
no data available
Number of animals or in vitro replicates:
3 male rabbits
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated eye of rabbit was washed with normal saline.
- Time after start of exposure: 24 hours

SCORING SYSTEM: Draize Method

TOOL USED TO ASSESS SCORE: Ophthalmoscope and fluorescent strips.
Irritation parameter:
cornea opacity score
Basis:
animal: #1,#2 and #3
Time point:
7 d
Score:
0
Max. score:
3
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
Untreated (Control Eye)
Irritation parameter:
iris score
Basis:
animal: #1,#2 and #3
Time point:
7 d
Score:
0
Max. score:
3
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
Untreated (Control Eye)
Irritation parameter:
conjunctivae score
Basis:
animal: #1,#2 and #3
Time point:
7 d
Score:
0
Max. score:
3
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
Untreated (Control Eye)
Irritation parameter:
chemosis score
Basis:
animal: #1,#2 and #3
Time point:
7 d
Score:
0
Max. score:
3
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
Untreated (Control Eye)
Irritation parameter:
cornea opacity score
Basis:
animal: #1,#2 and #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritation parameter:
iris score
Basis:
animal: #1,#2 and #3
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritation parameter:
conjunctivae score
Basis:
animal: #1,#2 and #3
Time point:
24/48/72 h
Score:
0.33
Max. score:
3
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritation parameter:
chemosis score
Basis:
animal: #1,#2 and #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
not specified
Remarks on result:
no indication of irritation
Remarks:
For treated eyes
Irritant / corrosive response data:
The following were observed in treated rabbits.
Observation at 1 hour after instillation of test item revealed: Cornea- No ulceration or opacity in all the animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) was observed in all the animals; Chemosis: Some swelling above normal (includes nictitating membranes) was observed in all the animals.

Observation at 24 hours after instillation of test item revealed: Cornea- No ulceration or opacity in all the animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) was observed in all the animals; Chemosis: No swelling (normal) was observed in all the animals.

At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 0% damage in all the animals.

Observation at 48 and 72 hours after instillation of test item revealed: Cornea- No ulceration or opacity in all the animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals. Conjunctivae - Blood vessels normal in all the animals; Chemosis: No swelling (normal) was observed in all the animals.

The individual mean score for animal nos. 1, 2 and 3 at 24, 48, 72 hours (at treated site) for corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 0.33, 0.00; 0.00, 0.00, 0.33, 0.00 and 0.00, 0.00, 0.33, 0.00 respectively.
Other effects:
Clinical Observation
No systemic toxicity was observed in treated rabbits during the experimental period

Mortality
No mortality was observed during the experimental period.

Body weight
Increase in body weights in all the treated animals at termination when compared to day 0 was observed.

Table 1: Individual Animal Eye Irritation Scores

In Treated area Dose: 0.1 ml of as such test item                                                 Sex: Male

Animal Numbers

1

2

3

Application Side

Right

Right

Right

Eye Reactions

At hour

At hour

At hour

*

1

24

48

72

*

1

 24

48

72

*

1

24

48

72

Corneal Opacity

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Area of Opacity

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Iris

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Conjunctiva

0

1

1

0

0

0

1

1

0

0

0

1

1

0

0

Chemosis

0

1

0

0

0

0

1

0

0

0

0

1

0

0

0

Corneal Damage%

0

./.

0

0

./.

0

0

./.

0

 

In Treated area Dose: Untreated                                                                             Sex: Male

Animal Numbers

1

2

3

Application Side

Right

Right

Right

Eye Reactions

At hour

At hour

At hour

*

1

24

48

72

*

1

 24

48

72

*

1

24

48

72

Corneal Opacity

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Area of Opacity

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Iris

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Conjunctiva

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Chemosis

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Corneal Damage%

0

./.

0

0

./.

0

0

./.

0

Key: *= Pre-exposure eye examination.

      ./.= Not Applicable

Eye Irritation Scores - Mean Values after 24, 48, 72 Hours (Treated eye)

            Animal No.

 Eye Reaction

1

2

3

Corneal Opacity

0.00

0.00

0.00

Iris

0.00

0.00

0.00

Conjunctiva

0.33

0.33

0.33

Chemosis

0.00

0.00

0.00

Eye Irritation Scores - Mean Values after 24, 48, 72 Hours (Untreated eye)

            Animal No.

 Eye Reaction

1

2

3

Corneal Opacity

0.00

0.00

0.00

Iris

0.00

0.00

0.00

Conjunctiva

0.00

0.00

0.00

Chemosis

0.00

0.00

0.00

Formula :

Mean Eye Irritation Score = Sum of the Individual Animal Score for eye reaction at 24, 48 and 72 hours

Number of the Observations (3)

Table 2 : Individual Animal Clinical Signs

Animal No.

Days (Post application observation)

0

1

2

3

1

1

1

1

1

2

1

1

1

1

3

1

1

1

1

Key: 1 = Normal

Table 3: Individual Animal Body Weight

Animal No.

Animal Body Weight (kg)

Prior to application

At termination

1

2.146

2.180

2

2.146

2.198

3

1.822

1.886

Key: kg = Kilogram

Interpretation of results:
other: not irritating
Conclusions:
The individual mean score for animal nos. 1, 2 and 3 at 24, 48, 72 hours (at treated site) for corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 0.33, 0.00; 0.00, 0.00, 0.33, 0.00 and 0.00, 0.00, 0.33, 0.00 respectively. Under the experimental conditions tested, the test chemical was considered to be Non Irritant to New Zealand White male rabbit eyes and is thus Classified as “Category- Not Classified as Eye Irritant” as per CLP criteria for Classification.
Executive summary:

Acute Eye Irritation/Corrosion Study of the test chemical in Rabbits, was performed as per OECD guideline no. 405. 3 male New Zealand White rabbits were used for the study. Rabbits free from injury of eye were selected for the study. The eyes of all the rabbits were examined 24 hours prior to treatment. One eye of each rabbit served as control and other as treated. Control eye was left untreated whereas; 0.1 ml of test chemical (as such) was instilled in the other (treated) eye of rabbits. The eye was observed at 1, 24, 48, 72 hours after test chemical instillation. Ophthalmoscope was used for scoring of eye lesions. In the initial test,0.1 ml of as such test chemical was instilled into the conjunctival sac of the right eye of animal no.1 whereas the left eye of the rabbit served as the control. As animal no. 1 showed no severe ocular lesions hence a confirmatory test was conducted on additional two rabbits (animal no. 2 and 3);0.1 ml of as such test chemical was instilled into the conjunctival sac of right eye of both the rabbits and left eye served as the control. Untreated eye of all the three rabbits was normal throughout the experimental period of 72 hours. The following grading scores were observed in treated eye of tested rabbits. Observation at 1 hour after instillation of test chemical revealed: Cornea- No ulceration or opacity in all the animals; Area of Opacity-Zero in all the animals; Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) was observed in all the animals; Chemosis: Some swelling above normal (includes nictitating membranes) was observed in all the animals. Observation at 24 hours after instillation of test chemical revealed: Cornea- No ulceration or opacity in all the animals; Area of Opacity-Zero in all the animals; Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) was observed in all the animals; Chemosis: No swelling (normal) was observed in all the animals. At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 0% damage in all the animals. Observation at 48 and 72 hours after instillation of test chemical revealed: Cornea- No ulceration or opacity in all the animals; Area of Opacity-Zero in all the animals; Iris: Normal in all the animals. Conjunctivae - Blood vessels normal in all the animals; Chemosis: No swelling (normal) was observed in all the animals. The individual mean score for animal nos. 1, 2 and 3 at 24, 48, 72 hours (at treated site) for corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 0.33, 0.00; 0.00, 0.00, 0.33, 0.00 and 0.00, 0.00, 0.33, 0.00 respectively. Under the experimental conditions tested, the test chemical was considered to be Non Irritant to New Zealand White male rabbit eyes and was thus Classified as "Not Classified ” as per CLP criteria for Classification.

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Remarks:
Read-across from supporting substance (surrogate)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is from peer reviewed journal.
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Principles of method if other than guideline:
An ocular irritation study was conducted for the test chemical on six New Zealand albino rabbits to assess its adverse effects on eye.
GLP compliance:
not specified
Species:
rabbit
Strain:
other: New Zealand albino
Details on test animals or tissues and environmental conditions:
Weight at study initiation: 1.9-3.1 kg
Vehicle:
not specified
Controls:
yes
Amount / concentration applied:
concentration applied :70%
Amount : 0.1 ml per dose
Duration of treatment / exposure:
7 days


Observation period (in vivo):
1,24,48, 72 hours and 7 days
Duration of post- treatment incubation (in vitro):
No Data Available
Number of animals or in vitro replicates:
6 rabbits
Details on study design:
TEST SITE
- Area of exposure: conjunctival sac of right eye of each animal.
SCORING SYSTEM: According to the grading and scoring system of Draize et al. (1944) in which a zero score indicates no irritation and the maximum score at any one scoring period is 110 (maximum irritation and damage to the cornea, iris, and conjunctiva).
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
7 d
Score:
0
Max. score:
110
Reversibility:
fully reversible within: 3 days
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
Slight ocular lesions were observed but all the effects were reversible within 3 days.
Other effects:
No Data Available

Table 1: Primary Irritation score in rabbits

Time Point

Primary Irritation Score

1 hour

12

24 hour

7.5

48 hours

4.6

72 hours

0

7 days

0

Interpretation of results:
other: Not irritating
Conclusions:
The mean irritation score of test chemical was observed to be 0.0 after 72 hours. Therefore, the test chemical was considered to be not irritating to the eyes of six New Zealand albino rabbits.
Executive summary:

An ocular irritation study was conducted for the test chemical on six New Zealand albino rabbits to assess its adverse effects on eye. The 0.1ml dose of 70% of the test chemical was instilled into the conjunctival sac of the right eye of each of six rabbit while the left eye of each animal served as the control. Grading for eye injury was made at 1 , 24, 48 and 72 hr. and at 7 days after treatment according to the grading and scoring system of Draize et al. (1944) in which a zero score indicates no irritation and the maximum score at any one scoring period is 110 (maximum irritation and damage to the cornea, iris, and conjunctiva). Slight ocular lesions were observed but all the effects were cleared within 3 days. Hence, the test chemical was considered to be not irritating to the eyes of six New Zealand albino rabbits.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin Irritation

Various studies have been reviewed to evaluate the dermal irritation potential of the test chemical. These include in vivo experimental studies performed on guinea pigs as well as humans for the test chemical.

A Preliminary irritation tests for test chemical were performed on male albino Dunkin/Hartley guinea pigs to determine the concentration for suitable the sensitization study. In the preliminary test, the guinea pigs were treated intradermally and dermally. The animals were intradermally injected with 0.1ml aliquots of a range of concentration of test substance (0.25%,0.5%, 1% or 2%) in 6% acetone/ 20% PEG 400 /0.01% Tween 80/ saline. During topical application, animals received a single dermal application of 10%, 25% or 50% test chemical in acetone using 8 mm saturated filter paper and 11 mm aluminum patch test cups which were applied to the shaved flank for 24 h. Reactions were assessed for irritation at 24 and 48 h after patch removal. Very slight erythema was observed at 0.25% and 0.5%. Barely perceptible erythema was observed at 10%, 25% or 50% concentrations. Since the test chemical induced cutaneous effects at these concentrations, the test chemical was considered to be slightly irritating to the skin of albino Dunkin/Hartley guinea pigs. Thus concentration of 0.5% was selected for the intradermal induction and a concentration of 50% the test chemical was selected for the topical induction application. Based on the observations and results, it was concluded that the test material was considered to be slightly irritating (Category 2) on male albino Dunkin/Hartley guinea pigs in the preliminary irritation tests performed for the sensitization study.

This result is supported by another Preliminary irritation test performed in guinea pigs to determine the suitable challenge concentration for the sensitization study of the test chemical. The animals received a single dermal application of 10%, 25% or 50% of the test chemical in acetone using 8 mm saturated filter paper and 11 mm aluminum patch test cups which were applied to the shaved flank for 24 h. Reactions were assessed for irritation at 24 and 48 h after patch removal. Barely perceptible erythema was observed at all concentrations. Since the test chemical did not induce any cutaneous effects at concentration of 10%, the test chemical was considered to be not irritating to the skin of male albino Dunkin/Hartley guinea pigs. Thus 10% was selected for the challenge application in sensitization study. Thus, based on all the observations and results, the test chemical was considered to be non-irritating at concentration of 10% on skin of male albino Dunkin/Hartley guinea pigs.

The key study is supported by a similar pre-test of an OET (Open Epicutaneous Test) was conducted on Himalayan white-spotted guinea pigs for test chemical for establishing the primary irritating threshold concentration of test material. Male and female outbred Himalayan white-spotted guinea pigs bred at the Institute of Biomedical Research Fullinsdof, Switzerland. The dose concentrations tested were 100%, 30%, 10%, 3%, 1%, or 0.3% A 0.025 ml aliquot of the test chemical was applied to a 2 cm 2 area on the clipped flank. The application site was left uncovered and reactions were read after 24 h. It was observed thata concentration of 0.1% was the lowest concentration to produce mild irritation after a single application and was selected as the minimal irritating concentration. Therefore, the test chemical was considered to be irritating at concentration of 0.03–100% on Himalayan white-spotted guinea pigs.

These results are further supported by a skin irritation test of the test chemical was conducted on Himalayan white-spotted guinea pigs prior to the induction of an open epicutaneous test (OET). Male and female outbred Himalayan white-spotted guinea pigs bred at the Institute of Biomedical Research Fullinsdof, Switzerland. The dose concentrations tested were 100%, 30%, 10%, 3%, 1%, or 0.3%. A 0.1 ml aliquot of neat test chemical and progressively diluted solutions of the test chemical were applied to a 8 cm2area on the clipped flanks of 6–8 guinea pigs/group. The applications were repeated daily for 21 days, using the same skin site. The sites were left uncovered and the reactions were read 24 h after each application. Minimal irritation was observed at concentration of 0.1%. Therefore, the test chemical was considered to be irritating at concentration of 0.03–100% on Himalayan white-spotted guinea pigs. Based on all the observations and results, it was concluded that the test chemical was considered to be irritating on Himalayan white-spotted guinea pigs at concentration 0.03–100%.

These results are also supported by a skin irritation test that was conducted on 5 volunteers as a part of maximization study to conduct the skin irritation potential of test chemical. The test chemical was

applied at concentration of 8% to normal sites on the backs of each volunteer for 48 h under occlusive condition and later observed for skin lesions. Since none of the volunteer developed any skin reaction. The test chemical was considered to be not irritating on skin of five healthy volunteers.

Eventhough there is evidence that the test chemical lacks the potential to cause dermal irritation to rabbit/guinea pig skin. But the remaining results indicate otherwise. Hence, considering the majority of the results, the test chemical can be concluded to be irritating to skin and classified under the category “Category 2” as per CLP Regulation.

 

Eye Irritation:

Various studies have been reviewed to evaluate the ocular irritation potential of the test chemical. These include experimental studies performed on rabbits for the test chemical. The results are as follows:

An acute Eye Irritation/Corrosion Study of the test chemical in Rabbits, was performed as per OECD guideline no. 405. 3 male New Zealand White rabbits were used for the study. Rabbits free from injury of eye were selected for the study. The eyes of all the rabbits were examined 24 hours prior to treatment. One eye of each rabbit served as control and other as treated. Control eye was left untreated whereas; 0.1 ml of test chemical (as such) was instilled in the other (treated) eye of rabbits. The eye was observed at 1, 24, 48, 72 hours after test chemical instillation. Ophthalmoscope was used for scoring of eye lesions. In the initial test,0.1 ml of as such test chemical was instilled into the conjunctival sac of the right eye of animal no.1 whereas the left eye of the rabbit served as the control. As animal no. 1 showed no severe ocular lesions hence a confirmatory test was conducted on additional two rabbits (animal no. 2 and 3);0.1 ml of as such test chemical was instilled into the conjunctival sac of right eye of both the rabbits and left eye served as the control. Untreated eye of all the three rabbits was normal throughout the experimental period of 72 hours. The following grading scores were observed in treated eye of tested rabbits. Observation at 1 hour after instillation of test chemical revealed: Cornea- No ulceration or opacity in all the animals; Area of Opacity-Zero in all the animals; Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) was observed in all the animals; Chemosis: Some swelling above normal (includes nictitating membranes) was observed in all the animals. Observation at 24 hours after instillation of test chemical revealed: Cornea- No ulceration or opacity in all the animals; Area of Opacity-Zero in all the animals; Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) was observed in all the animals; Chemosis: No swelling (normal) was observed in all the animals. At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 0% damage in all the animals. Observation at 48 and 72 hours after instillation of test chemical revealed: Cornea- No ulceration or opacity in all the animals; Area of Opacity-Zero in all the animals; Iris: Normal in all the animals. Conjunctivae - Blood vessels normal in all the animals; Chemosis: No swelling (normal) was observed in all the animals. The individual mean score for animal nos. 1, 2 and 3 at 24, 48, 72 hours (at treated site) for corneal opacity, iris, conjunctiva and chemosis were found 0.00, 0.00, 0.33, 0.00; 0.00, 0.00, 0.33, 0.00 and 0.00, 0.00, 0.33, 0.00 respectively. Under the experimental conditions tested, the test chemical was considered to be Non Irritant to New Zealand White male rabbit eyes and was thus Classified as "Not Classified ” as per CLP criteria for Classification.

This result is supported by ocular irritation study, was conducted for the test chemical on six New Zealand albino rabbits to assess its adverse effects on eye. The 0.1ml dose of 70% of the test chemical was instilled into the conjunctival sac of the right eye of each of six rabbit while the left eye of each animal served as the control. Grading for eye injury was made at 1 , 24, 48 and 72 hr. and at 7 days after treatment according to the grading and scoring system of Draize et al. (1944) in which a zero score indicates no irritation and the maximum score at any one scoring period is 110 (maximum irritation and damage to the cornea, iris, and conjunctiva). Slight ocular lesions were observed but all the effects were cleared within 3 days. Hence, the test chemical was considered to be not irritating to the eyes of six New Zealand albino rabbits.

Based on the available results and applying the weight of evidence approach the test chemical can be considered to be not irritating to the eyes and can be classified under the category “Not Classified” as per CLP Regulation.

Justification for classification or non-classification

Eventhough there is evidence that the test chemical lacks the potential to cause dermal irritation to rabbit/guinea pig skin. But the remaining results indicate otherwise. Hence, considering the majority of the results, the test chemical can be concluded to be irritating to skin and classified under the category “Category 2” as per CLP Regulation.

Based on the available results and applying the weight of evidence approach the test chemical can be considered to be not irritating to the eyes and can be classified under the category “Not Classified” as per CLP Regulation.