Registration Dossier

Administrative data

Description of key information

Skin Irritation:

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. 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.

 

Eye Irritation:

Acute Eye Irritation/Corrosion Study of Test chemical in Rabbits, was performed in an OECD GLP laboratory (Sustainability Support Services (Europe) has letter of access).The study was performed according to OECD 405 Guidelines. The individual mean score for Animal Nos. 1, 2 and 3 at 24, 48, 72 hours for Corneal opacity, iris, conjunctiva, chemosis were found  0.00, 0.00, 1.00; 0.33; 0.00, 0.00; 1.00, 0.00 and 0.00, 0.00, 1.00, 0.00,  respectively. Hence, under the experimental conditions, the test chemical is “Non Irritant” to New Zealand White male rabbit eyes.

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 reports
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
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:
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.
Other effects / acceptance of results:
no data available
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.

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.


 

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.

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

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye 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
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Principles of method if other than guideline:
Eye irritation study was performed for test chemical.
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 RABBI ROOF Animal Facility, A. P., India. (CPCSEA Reg. No. 1655/PO/b/12/CPCSEA).
- Age at study initiation:4.5 to 5.5 Months (Approximately)
- Health Status: Healthy young adult
- Weight at study initiation: Minimum: 1.696 kg and Maximum: 1.942 kg (Prior to Treatment)
- 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.
- 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 6 days (Animal No.-1) and 10 days (Animal No. 2 and 3) prior to the application of the test item
- 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: 19.50 °C and Maximum: 22.20 °C
- Humidity (%): Minimum: 41.20 % and Maximum: 59.20 %
- Air changes (per hr): More than 12 changes per hour
- Photoperiod (hrs dark / hrs light): 12:12
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
0.1 ml
Duration of treatment / exposure:
24 hours
Observation period (in vivo):
Animals were observed at 24 hours intervals for 72 hours.
Duration of post- treatment incubation (in vitro):
no data available
Number of animals or in vitro replicates:
3 males
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes
- Time after start of exposure: After 24 hours observation period, the treated eye of rabbit was washed with normal saline

SCORING SYSTEM: Draize method

TOOL USED TO ASSESS SCORE: hand-slit lamp / biomicroscope / fluorescein:The area of corneal epithelial cell damage was scored using ophthalmoscope (Ri-Mini2, 5 V-guru; Model No. 3002).
Other effects / acceptance of results:
no data available
Irritation parameter:
cornea opacity score
Basis:
animal: 1,2,3
Time point:
24/48/72 h
Score:
0
Reversibility:
not specified
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal: 1,2,3
Time point:
24/48/72 h
Score:
0
Reversibility:
not specified
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal: 1,2,3
Time point:
24/48/72 h
Score:
1
Reversibility:
not specified
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal: 1,2,3
Time point:
24/48/72 h
Score:
0.33
Reversibility:
not specified
Remarks on result:
no indication of irritation
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 3 animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals. Conjunctivae - Some blood vessels definitely hyperaemic (injected) in animal no. 1 and 2 whereas animal no.3 showed no ocular lesion; Chemosis: No swelling (Normal) were observed in all the animals.
Observation at 24 hours after instillation of test item revealed: Cornea- No ulceration or opacity in all 3 animals; Area of Opacity- Zero in all the animals; Iris: Normal in all the animals. Conjunctivae - Diffuse, crimson color; individual vessels not easily discernible was observed in all the treated animals; Chemosis: Some swelling above normal (includes nictating membranes) was observed in animal number 1 and no swelling was observed in animal number 2 and 3.
At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 40 % and 15 % damage in animal no 1 and 2 whereas no damage was seen in animal no. 3.
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 were normal in all the animals; Chemosis: No swelling was observed in all the animals.
The individual mean score for Animal Nos. 1, 2 and 3 at 24, 48, 72 hours for Corneal opacity, iris, conjunctiva, chemosis were found 0.00, 0.00, 1.00; 0.33; 0.00, 0.00; 1.00, 0.00 and 0.00, 0.00, 1.00, 0.00, 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 were weighed on test day 0 (prior to application) and at termination. The body weights were marginally increased in animal number 1, 2 and 3 when compared with pre-treatment body weights.

Table 1 : Individual Animal Eye Irritation Score

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

1.00

1.00

1.00

Chemosis

0.33

0.00

0.00

 

 

Formula :

 

Mean Eye Irritation Score =

                          Sum of the Individual Animal Score for eye reactionat24, 48 and 72 hours

Number of the Observations (3)

Table 2 : Individual AnimalClinicalSigns

 

Sex:Male

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

Sex :Male

Animal No.

Animal Body Weight (kg)

Prior to application

At termination

1

1.696

1.812

2

1.942

1.982

3

1.800

1.826

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 for Corneal opacity, iris, conjunctiva, chemosis were found  0.00, 0.00, 1.00; 0.33; 0.00, 0.00; 1.00, 0.00 and 0.00, 0.00, 1.00, 0.00,  respectively.
Hence, under the experimental conditions, the test chemical is “Non Irritant” to New Zealand White male rabbit eyes
Executive summary:

Acute Eye Irritation/Corrosion Study of Test chemical in Rabbits, was performed in an OECD GLP laboratory (Sustainability Support Services (Europe) has letter of access).The study was performed according to OECD 405 Guidelines. 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 item (as such) was instilled in the other (treated) eye of rabbits. The eye was observed at 1, 24, 48, 72 hours after test item instillation. Ophthalmoscope was used for scoring of eye lesions. In the initial test,0.1 ml of test item was applied into the conjunctival sac of the right eye of Animal No.1. The left eye of the rabbit served as the control. Animal No. 1 presented ocular lesions at 1 hour observation period. Hence the confirmatory test was conducted on additional two rabbits (Animal No. 2 and 3);0.1 ml of test item was instilled into the conjunctival sac of right eye and left eye served as the control. Ocular lesions were observed at 1, 24 and 48 hour in Animal Number 2 whereas in Animal Number 3 ocular lesions presented only at 1 hour observation period. Untreated eye of the treated 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 item revealed: Cornea-No ulceration or opacity in all 3 animals; Area of Opacity-Zero in all the animals;Iris:Normal in all the animals. Conjunctivae -Some blood vessels definitely hyperaemic (injected) in animal no. 1 and 2 whereas animal no.3 showed no ocular lesion;Chemosis:No swelling (Normal) were observed in all the animals. Observation at 24 hours after instillation of test item revealed: Cornea-No ulceration or opacity in all 3 animals; Area of Opacity-Zero in all the animals;Iris:Normal in all the animals.Conjunctivae -Diffuse, crimson color; individual vessels not easily discernible was observed in all the treated animals;Chemosis:Some swelling above normal (includes nictating membranes) was observed in animal number 1 and no swelling was observed in animal number 2 and 3. At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 40 % and 15 % damage in animal no 1 and 2 whereas no damage was seen in animal no. 3. 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 were normal in all animals; Chemosis: No swelling was observed in all the animals. Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbits was observed at 72 hours. The individual mean score for Animal Nos. 1, 2 and 3 at 24, 48, 72 hours for Corneal opacity, iris, conjunctiva, chemosis were found  0.00, 0.00, 1.00; 0.33; 0.00, 0.00; 1.00, 0.00 and 0.00, 0.00, 1.00, 0.00,  respectively. Hence, under the experimental conditions, Test chemical is “Non Irritant” to New Zealand White male rabbit eyes.

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin Irritation:

 

Various studies were performed on humans, rabbits to assess the dermal irritation potential of test chemical which have been summarized as follows:

 

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.

 

 

This result is supported by the experimental study designed to investigate the irritation effects of test chemical. The test chemical was applied once to intact skin under a semi-occlusive dressing which enhances penetration and interaction of the test materials and the skin. Diethyl Phthalate, Cyclamen Aldehyde and Geraniol are standard perfumery controls used in the study.0.5 ml of test chemical and standard solution (0.2 gm for solid samples) are applied to the clipped dorsum of 8 New Zealand White rabbits for 4 hours under a semi-occlusive patch.  The irritation reactions were assessed immediately after removal of the patch and after 24, 48 and 72 hours. A single treatment of the perfumery material produced a slight/moderate effect 24 hours after treatment. The level of reaction subsided to a generally slight reaction 72 hours after treatment. The response was significantly greater than that produced by Diethyl Phthalate which showed no response after 72 hours and consistently less than that produced by Cyclamen Aldehyde and Geraniol which produced a generally moderate reaction. Under the conditions of this test the test chemical produced slight/ moderate skin irritation. It was somewhat less effective than two widely used perfumery ingredients which are routinely used as standard markers in this type of test. The test substance would not be expected to cause undue skin irritation during normal factory handling.

 

These results are further supported by the experimental study conducted for test chemical in humans. 11 adult male and female volunteers were chosen for the test. Undiluted (full strength) test substance was applied to an area of approximately 1cm in diameter on the dermis of the inner portion of lower arm. After application, the test site was covered with adhesive bandage. The readings were recorded at 24 hour intervals for 5 days. Undiluted chemical did not produce any signs of irritation. Hence, the test substance can be considered to be not irritating to skin.

 

The above results are also supported by the experimental study conducted in humans. The study was conducted to obtain practically non irritating perfumes for soaps, the primary irritation tests were performed on male and female human volunteers by closed patch tests. The test chemical was also tested in this study along with various other fragrance chemicals. 3 patch tests were performed at different concentrations of test chemical on human volunteers. The vehicles used for the patch tests were- Test A: in vaselin or unguentum hydrophilicum; Test B: in unguentum simplex or unguentum hydrophilicum; Test C: in 99% ethanol or non-irritative cream base. The concentration of the test material applied to the test sites (test 1: Small of back, Test B,C: Upper inside of arm)were20%, 2% and 0.2% respectively in 29,30 and 42 humans (male and female). The volunteers were exposed to the test chemical for 48 hours, 24 -72 hours, 24 -48 hours respectively. The volunteers were assessed for signs of erythema and assessed according to the scale: -= No reaction;±=Slight Erythema; += Erythema and dropsical swelling; ++= Erythema and dropsical swelling; +++= Erythema, papules and vesicles. In all of the tests conducted, the chemical did not cause any irritation on the skin of the human volunteers. Based on the results obtained in the 3 closed patch tests on human volunteers, it was considered that the test substance was not irritating to skin and can be used as a perfume for soaps.

 

All these studies lead to a conclusion that Test chemical is indeed not irritating to skin. Hence, comparing the above annotations with the criteria of CLP regulation, Test chemical can be classified under the category “Not Classified”.

 

 

 

Eye Irritation:

 

Various studies were performed on rabbits to assess the ocular irritation potential of test chemical which have been summarized as follows:

 

Acute Eye Irritation/Corrosion Study of Test chemical in Rabbits, was performed in an OECD GLP laboratory (Sustainability Support Services (Europe) has letter of access).The study was performed according to OECD 405 Guidelines. 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 item (as such) was instilled in the other (treated) eye of rabbits. The eye was observed at 1, 24, 48, 72 hours after test item instillation. Ophthalmoscope was used for scoring of eye lesions. In the initial test,0.1 ml of test item was applied into the conjunctival sac of the right eye of Animal No.1. The left eye of the rabbit served as the control. Animal No. 1 presented ocular lesions at 1 hour observation period. Hence the confirmatory test was conducted on additional two rabbits (Animal No. 2 and 3);0.1 ml of test item was instilled into the conjunctival sac of right eye and left eye served as the control. Ocular lesions were observed at 1, 24 and 48 hour in Animal Number 2 whereas in Animal Number 3 ocular lesions presented only at 1 hour observation period. Untreated eye of the treated 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 item revealed: Cornea-No ulceration or opacity in all 3 animals; Area of Opacity-Zero in all the animals;Iris:Normal in all the animals. Conjunctivae -Some blood vessels definitely hyperaemic (injected) in animal no. 1 and 2 whereas animal no.3 showed no ocular lesion;Chemosis:No swelling (Normal) were observed in all the animals. Observation at 24 hours after instillation of test item revealed: Cornea-No ulceration or opacity in all 3 animals; Area of Opacity-Zero in all the animals;Iris:Normal in all the animals.Conjunctivae -Diffuse, crimson color; individual vessels not easily discernible was observed in all the treated animals;Chemosis:Some swelling above normal (includes nictating membranes) was observed in animal number 1 and no swelling was observed in animal number 2 and 3. At 24 hours observation the rabbits were examined for corneal epithelium cell damage using sodium fluorescein strips and noticed 40 % and 15 % damage in animal no 1 and 2 whereas no damage was seen in animal no. 3. 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 were normal in all animals; Chemosis: No swelling was observed in all the animals. Under the experimental conditions tested, eye irritation and reversibility of effects on eyes of rabbits was observed at 72 hours. The individual mean score for Animal Nos. 1, 2 and 3 at 24, 48, 72 hours for Corneal opacity, iris, conjunctiva, chemosis were found  0.00, 0.00, 1.00; 0.33; 0.00, 0.00; 1.00, 0.00 and 0.00, 0.00, 1.00, 0.00,  respectively. Hence, under the experimental conditions, Test chemical is “Non Irritant” to New Zealand White male rabbit eyes.

 

 

 

The above result was supported by an eye irritation test conducted in three rabbits to assess the ocular irritation potential of test chemical. A 0.1 ml aliquot of 100% and 5% of chemical in DEP was instilled into one eye of each of three rabbits with no further treatment. The untreated eye served as a control. Observations for irritation were made immediately after instillation and again at 1, 2, 4, 24, 48 and 72 h after instillation. Very slight conjunctival irritation was observed in all three rabbits at instillation, which cleared by 24 hours .The instillation of test chemical did not cause any ocular irritation in 3 rabbits after 72 hours of observation. Hence the test chemical can be considered as a non-irritant to rabbit eyes.

 

 

The above results were further supported by another ocular irritation study conducted in rabbits for test chemical. The study was performed according to Draize method. Neat test chemical (0.1 ml) was instilled in to the eyes of 3 rabbits. The untreated eyes served as controls. Animals were observed for effects after 1, 2, 4, 24, 48, and 72 h instillation of the test chemical. Fluorescein was used to check corneal damage. Neat test chemical produces very slight conjunctival irritation in all three rabbits at 0, 1, 2, and 4 hours. No other effects were observed. Hence, the test chemical was considered not irritating to rabbit eyes.

 

The results obtained from these studies lead to a conclusion that Test chemical is indeed not irritating to eye. Hence, comparing the above annotations with the criteria of CLP regulation, Test chemical can be classified under the category “Not Classified”.

 

Justification for classification or non-classification

The skin and eye irritation potential of test chemical was observed in various studies. The results obtained from these studies indicate that the chemical is not likely to cause skin and eye irritation. Hence the test chemical can be classified under the category “Not Classified” for skin and eye as per CLP.