2-ethoxybenzoic acid

Endpoint summary

Description of key information

Skin Irritation:

The overall irritation score of the substance from in-vivo and in-vitro studies were determined to be 0 and no erythema and edema (skin irritation) were determine at the end of 14 days observation period after patch removal. Hence, it was concluded that the test chemical was Non-Irritating to the skin of female Sprague Dawley rats under the experimental conditions tested and classified as “Category- Not Classified” as per CLP Classification.

Eye Irritation:

By applying the weight of evidence approach, the test chemical can be considered to be irritating to eyes. Comparing the annotations with criteria of CLP regulation, the test chemical can be classified under the category "Category 2".

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:

comparable to guideline study

Justification for type of information:

data is from experimental reports

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 402 (Acute Dermal Toxicity)

Principles of method if other than guideline:

To determine the dermal reaction profile of the test chemical in Sprague Dawley rats

GLP compliance:

yes

Specific details on test material used for the study:

- Name of test material (as cited in study report): 2 Ethoxybenzoic acid
- Molecular formula : C9H10O3
- Molecular weight : 166.175 g/mol
- Substance type: Organic
- Physical state: Liquid
Laboratory Test Item Code : TAS/122/057
Manufacturing Date : November; 2016
Expiry Date : October; 2019
Consistency : Liquid
Activity (Clinical Indication) : Industrial chemical
Safety Precautions : Safety precautions included use of protective clothing, gloves, masks and eye protection (glasses).
Storage Condition : Ambient Temperature
Preparation of Test Item : The test item was used undiluted
Batch Number : EB-22/16-17

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: National Institute of Biosciences, Pune.
- Sex: Female. Females were nulliparous and non-pregnant
- Age: young adult, 8-10 weeks old
- Weight at study initiation: The weight range of approximately 220.2 to 240.6 grams at initiation of dosing.
Body weights at the start :
Female
Mean : 231.34 g (= 100 %)
Minimum : 220.2 g (- 4.82 %)
Maximum : 240.6 g (+ 4.00 %)
Total No. of animals : 5
- Identification: Each rat was individually identified by the cage number.
- Fasting period before study: No data available
- Housing: The rats were individually housed in polycarbonate cages with paddy husk as bedding.
- Diet (e.g. ad libitum): Rodent feed supplied by the Nutrivet Life Sciences, Pune, was provided ad libitumfrom individual feeders.
- Water (e.g. ad libitum): Water was provided ad libitum from individual bottles attached to the cages. All water was from a local source and passed through the reverse osmosis membrane before use.
- Acclimation period: 5 days.

ENVIRONMENTAL CONDITIONS

- Temperature (°C): 19.2 to 21.8 degree centigrade.
- Humidity (%): 56.0% to 59.1%
- Air changes (per hr): Ten to fifteen air changes per hour.
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room.

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

not specified

Amount / concentration applied:

Dose range finding study:200 mg/kg, 1000 mg/kg and 2000 mg/kg
Main Study: 2000 mg/kg

Duration of treatment / exposure:

24 hours

Observation period:

14 days

Number of animals:

5 female

Details on study design:

TEST SITE
- Area of exposure: Trunk (dorsal surface and sides from scapular to pelvic area)
- % coverage: Approximately 10% of the body surface area.
- Type of wrap if used: Porous gauze dressing and non-irritating tape.
REMOVAL OF TEST SUBSTANCE
- Washing (if done): Distilled water was used to remove residual test item.
OBSERVATION TIME POINTS
(indicate if minutes, hours or days) : Dermal reaction was observed daily for study period of 14 days.
SCORING SYSTEM: Draize Method.

Irritation parameter:

erythema score

Basis:

mean

Time point:

14 d

Score:

0

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

mean

Time point:

14 d

Score:

0

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Evaluation of Dermal Reaction

Sex : Female
Dose Range Finding Study:
Group I : Animal treated at the dose level of 200 mg/kg body weight did not result in any skin reaction during the study period of 14 days.
Group I : Animal treated at the dose level of 1000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.
Group I : Animal treated at the dose level of 2000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.
Main Study:
Group II : Animals treated at the dose level of 2000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.

Other effects:

Body Weight
Sex : Female
Dose Range Finding Study:
Group I (200 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 6.09% and 11.30% respectively.
Group I (1000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 6.99% and 12.81% respectively.
Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 5.64% and 10.46% respectively.
Main Study:
Group II (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 3.21% and 7.29% respectively
Clinical Signs of Toxicity and Mortality
Sex : Female
Dose Range Finding Study:
Group I : Animal treated at the dose level of 200 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days. The animal survived through the study period of 14 days.
Group I : Animal treated at the dose level of 1000 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days. The animal survived through the study period of 14 days.
Group I : Animal treated at the dose level of 2000 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days. The animal survived through the study period of 14 days.
Main Study:
Group II : Animals treated at the dose level of 2000 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days. All animals survived through the study period of 14 days.
Gross Pathological Findings
Gross pathological examination did not reveal any abnormalities in animals from 200 mg/kg, 1000 mg/kg and 2000 mg/kg dose groups from dose range finding study and main study sacrificed terminally.

Individual Animal -Clinical Signs of Toxicity and Mortality

 

Laboratory Test Item Code :TAS/122/057

Test System : Sprague Dawley Rat

Sex : Female

Finding Study:

Group No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days From - to	Mortality
I	200	No clinical signs observed	1	1	Day 0 - Day 14	0

 

Group No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days From - to	Mortality
I	1000	No clinical signs observed	1	2	Day 0 - Day 14	0

 

Group No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days From - to	Mortality
I	2000	No clinical signs observed	1	3	Day 0 - Day 14	0

 

Main Study:

Group No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days From - to	Mortality
II	2000	No clinical signs observed	2	4	Day 0 - Day 14	0
				5	Day 0 - Day 14	0

 

Individual Animal - Evaluation of Dermal Reaction

 

Laboratory Test Item Code :TAS/122/057

Test System : Sprague Dawley Rat

Sex : Female  

Finding Study:

Group : I                                                                      Dose  : 200 mg/kg body weight

Animal	Dermal	D A Y S
No.	Reaction	0	1	2	3	4	5	6	7	8	9	10	11	12	13	14
1	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0

 

Group : I                                                                    Dose  : 1000 mg/kg body weight

Animal	Dermal	D A Y S
No.	Reaction	0	1	2	3	4	5	6	7	8	9	10	11	12	13	14
2	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0

 

Group : I                                                                    Dose  : 2000 mg/kg body weight

Animal	Dermal	D A Y S
No.	Reaction	0	1	2	3	4	5	6	7	8	9	10	11	12	13	14
3	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0

 

Main Study:

Group : II                                                                   Dose  : 2000 mg/kg body weight

Animal	Dermal	D A Y S
No.	Reaction	0	1	2	3	4	5	6	7	8	9	10	11	12	13	14
4	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
5	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0

 

 

Summary of Evaluation of Dermal Reaction

 

Laboratory Test Item Code :TAS/122/057

Test System : Sprague Dawley Rat

Sex : Female

Finding Study:

Group  No.	Dose mg/kg	Dermal Reaction	Total Number of Animals	Animal Nos.	Period of signs in days  From - to
I	200	No dermal reaction observed	1	1	Day 0 - Day 14

 

Group  No.	Dose mg/kg	Dermal Reaction	Total Number of Animals	Animal Nos.	Period of signs in days  From - to
I	1000	No dermal reaction observed	1	2	Day 0 - Day 14

 

Group  No.	Dose mg/kg	Dermal Reaction	Total Number of Animals	Animal Nos.	Period of signs in days  From - to
I	2000	No dermal reaction observed	1	3	Day 0 - Day 14

 

Main Study:

Group  No.	Dose mg/kg	Dermal Reaction	Total Number of Animals	Animal Nos.	Period of signs in days  From - to
II	2000	No dermal reaction observed	2	4, 5	Day 0 - Day 14

Mean Body Weight and Percent Body Weight Gain (g)

 

Laboratory Test Item Code :TAS/122/057

Test System : Sprague Dawley Rat

Sex : Female

Finding Study:

Group No.	Dose (mg/kg body weight)		Body weight Day 0	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
I	200	Mean	224.8	238.5	6.09	250.2	4.91	11.30
		± SD	-	-	-	-	-	-

 

Group No.	Dose (mg/kg body weight)		Body weight Day 0	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
I	1000	Mean	220.2	235.6	6.99	248.4	5.43	12.81
		± SD	-	-	-	-	-	-

 

Group No.	Dose (mg/kg body weight)		Body weight Day 0	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
I	2000	Mean	232.4	245.5	5.64	256.7	4.56	10.46
		± SD	-	-	-	-	-	-

 

Main Study:

Group No.	Dose (mg/kg body weight)		Body weight Day 0	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
II	2000	Mean	239.65	247.35	3.21	257.15	3.93	7.29
		± SD	1.34	6.86	2.28	12.94	2.35	4.80

 

Summary of Gross Pathological Findings

 

Laboratory Test Item Code :TAS/122/057

Test System : Sprague Dawley Rat

Sex : Female

Finding Study:

Group No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
I	200	1	TS	No abnormality detected

 

Group No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
I	1000	2	TS	No abnormality detected

 

Group No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
I	2000	3	TS	No abnormality detected

 

                    Main Study:

Group No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
II	2000	4, 5	TS	No abnormality detected

 

                     TS = Terminal Sacrifice

 

Interpretation of results:

other: not irritating

Conclusions:

The overall irritation score of the substance was determined to be 0 and no erythema and edema (skin irritation) were found at the end of 14 days observation period after patch removal.
Hence, it was concluded that the test chemical was Non-Irritating to the skin of Sprague Dawley rats under the experimental conditions tested and classified as “Category-Not Classified” as per CLP Classification.

Executive summary:

A study was designed and conducted to determine the dermal reaction profile of the test chemical in Sprague Dawley rats. The study was performed as per OECD Guidelines 402 and complying to the GLP procedures. 5 female young adult, non-pregnant rats were used for the study. The animals were kept in their cages for at least 5 days prior to administration for acclimatization to the laboratory condition and after acclimatization period, animals were randomly selected.

Approximately 24 hours before application, the hair of each rat was closely clipped from the trunk (dorsal surface and sides from scapular to pelvic area) with an electric clipper, so as to expose at least 10% of the body surface area. The test item was applied directly onto the exposed skin of the animal, taking care to spread the test item evenly over the entire area of approximately 10% of the total body surface area or as much of the area as can reasonably be covered. The test item was held in contact with the skin using a porous gauze dressing and non irritating tape around the animal to cover the exposure site for first 24 hours exposure period. Elizabethan collar was placed on each animal for first 24 hours after application of the test item. These collars prevent ingestion of test item. Following 24 hours of exposure, the wrapping was removed and the test site wiped free of excess test item. Distilled water was used to remove residual test item.

A single dose of 200 mg/kg body weight of the test item was administered to 1 female animal. No death or clinical signs of toxicity was observed during first 48 hours, hence, additional 1 female animal was administered at the dose of 1000 mg/kg body weight. Administration of 1000 mg/kg body weight did not reveal any clinical signs of toxicity or death during first 48 hours, hence, additional 1 female animal was administered at the dose of 2000 mg/kg body weight. Administration of 2000 mg/kg body weight did not reveal any clinical signs of toxicity or death during first 48 hours.

As the dose range finding study revealed no mortality or clinical signs at the maximum dose of 2000 mg/kg, the main study was initiated with two additional animals. The animals were administered with a dose of 2000 mg/kg body weight in sequential manner at 48 hours intervals. Animals from dose range finding study treated at the dose levels of 200 mg/kg, 1000 mg/kg and 2000 mg/kg and animals from main study treated at the dose level of 2000 mg/kg exhibited normal body weight gain and revealed no clinical signs of toxicity or mortality during the study period of 14 days. Gross pathological examination did not reveal any abnormalities attributable to the treatment.

The overall irritation score of the substance was determined to be 0 and no erythema and edema (skin irritation) were found at the end of 14 days observation period after patch removal.

Hence, it was concluded that the test chemical was Non-Irritating to the skin of female Sprague Dawley rats under the experimental conditions tested and classified as “Category- Not Classified” as per CLP Classification.

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:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Weight of evidence approach based on various test chemicals

Justification for type of information:

Weight of evidence approach based on various test chemicals

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: Weight of evidence approach based on various test chemicals

Principles of method if other than guideline:

Weight of evidence approach based on various test chemicals

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material (as cited in study report): 2 Ethoxybenzoic acid
- Molecular formula: C9H10O3
- Molecular weight: 166.175 g/mol
- Substance type: Organic
- Physical state: Liquid

Species:

rabbit

Strain:

New Zealand White

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

1,2. 100 mg
3. 0.005 ml

Duration of treatment / exposure:

single expsoure

Observation period (in vivo):

1,2. 1, 24, 48, 72, 96 hours and 7, 14, 21 days
3. 24 hours

Duration of post- treatment incubation (in vitro):

no data available

Number of animals or in vitro replicates:

1,2. 3
3. 5

Details on study design:

Weight of evidence approach based on test chemicals

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

21 d

Reversibility:

not specified

Remarks on result:

positive indication of irritation

Irritant / corrosive response data:

Signs of irritation observed

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

By applying the weight of evidence approach, the test chemical can be considered to be irritating to eyes. Comparing the annotations with criteria of CLP regulation, the test chemical can be classified under the category "Category 2".

Executive summary:

The eye irritation potential of the test chemical was assessed based on the available results from the various test chemicals.

The primary eye irritation potential of the test chemicals was evaluated according to Draize method.

3 female Japanese White rabbits (2.0-2.2 kg) were used for the study. The chemical (if solid= 100 mg, if liquid = 0.1ml) was instilled into the conjunctival sac of the left eye.Right eye (untreated eye) served as blank control. The eyes were examined and the grade of ocular lesions was recorded at 1, 24, 48, 72, 96 hours and 7,14,21 days after administration of test chemical. Corneal opacity, erythema, chemosis, secreta and iritis were recorded and classified under Draize method (Draize et.al, 1944).The sum of values, recorded for cornea, conjunctiva and iris was divided by the number of observation times and the average scores (Draize scores) were used as grades of ocular irritation potential of the chemical

The maximum value of Draize score are as follows: Cornea =80, Conjunctiva = 20 and Iris = 5

The average Draize scores after 21 days were as follows: CORNEA = 54.1/80, iris = could not be determined, conjunctiva = 10.3/20.

The test chemical induced severe eye irritation not recovering within 21 days of treatment.

Hence the test chemical can be assessed as irritating to rabbit eyes.             

This is supported by the results of the same study performed for the other test chemical. The average Draize scores after 21 days were as follows: CORNEA = 33.9/80, iris = 0.6, conjunctiva = 5.1.

The test chemical caused moderate eye irritation recovering within 21 days of treatment. Hence, the test chemical can be considered irritating to eyes.

 

The above Draize test results are further supported by another rabbit eye irritation test conducted in 5 healthy albino rabbits to assess the irritation potency of the test chemical. A 0.005 ml aliquot of neat test chemicals was applied to the center of the cornea while the lids were retracted. One minute later the lids were released. The eyes were examined 18–24 h later in strong diffuse daylight and then stained with fluorescein. The test chemical was assigned grade 3 – necrosis since 13–37% of the cornea was visible after fluorescein staining.

Therefore, the test chemical can be considered to be irritating to rabbit eyes.

By applying the weight of evidence approach, the test chemical can be considered to be irritating to eyes. Comparing the annotations with criteria of CLP regulation, the test chemical can be classified under the category "Category 2".

 

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin Irritation:

Various studies have been summarized to determine the degree of dermal irritation caused by the test chemical in living organisms. These studies include in-vivo experimental results obtained in rats, rabbits as well as humans.           

A study was designed and conducted to determine the dermal reaction profile of the test chemical in Sprague Dawley rats. The study was performed as per OECD Guidelines 402 and complying to the GLP procedures. 5 female young adult, non-pregnant rats were used for the study. The animals were kept in their cages for at least 5 days prior to administration for acclimatization to the laboratory condition and after acclimatization period, animals were randomly selected.

Approximately 24 hours before application, the hair of each rat was closely clipped from the trunk (dorsal surface and sides from scapular to pelvic area) with an electric clipper, so as to expose at least 10% of the body surface area. The test item was applied directly onto the exposed skin of the animal, taking care to spread the test item evenly over the entire area of approximately 10% of the total body surface area or as much of the area as can reasonably be covered. The test item was held in contact with the skin using a porous gauze dressing and non irritating tape around the animal to cover the exposure site for first 24 hours exposure period. Elizabethan collar was placed on each animal for first 24 hours after application of the test item. These collars prevent ingestion of test item. Following 24 hours of exposure, the wrapping was removed and the test site wiped free of excess test item. Distilled water was used to remove residual test item.

A single dose of 200 mg/kg body weight of the test item was administered to 1 female animal. No death or clinical signs of toxicity was observed during first 48 hours, hence, additional 1 female animal was administered at the dose of 1000 mg/kg body weight. Administration of 1000 mg/kg body weight did not reveal any clinical signs of toxicity or death during first 48 hours, hence, additional 1 female animal was administered at the dose of 2000 mg/kg body weight. Administration of 2000 mg/kg body weight did not reveal any clinical signs of toxicity or death during first 48 hours.

As the dose range finding study revealed no mortality or clinical signs at the maximum dose of 2000 mg/kg, the main study was initiated with two additional animals. The animals were administered with a dose of 2000 mg/kg body weight in sequential manner at 48 hours intervals. Animals from dose range finding study treated at the dose levels of 200 mg/kg, 1000 mg/kg and 2000 mg/kg and animals from main study treated at the dose level of 2000 mg/kg exhibited normal body weight gain and revealed no clinical signs of toxicity or mortality during the study period of 14 days. Gross pathological examination did not reveal any abnormalities attributable to the treatment.

The overall irritation score of the substance was determined to be 0 and no erythema and edema (skin irritation) were found at the end of 14 days observation period after patch removal. Hence, it was concluded that the test chemical was Non-Irritating to the skin of female Sprague Dawley rats under the experimental conditions tested and classified as “Category- Not Classified” as per CLP Classification.

The dermal irritation potential of test article was determined according to the OECD 439 test guideline followed for this study. The MatTek EpiDerm™ model was used to assess the potential dermal irritation of the test article by determining the viability of the tissues following exposure to the test article via MTT. Tissues were exposed to test article and controls for ~one hour, followed by a 42 hour post-exposure recovery period. The viability of each tissue was determined by MTT assay. The MTT data show the assay quality controls were met as the OD of the negative control tissues was between 1.195 and 1.430. Also, the positive control, 5% sodium dodecyl sulfate (SDS), reduced tissue viability to 4.5% of negative control and the standard deviation (SD) calculated from individual percent tissue viabilities of the test article exposed replicates was 3 passing the acceptance criteria. The mean of OD  for test chemical was determined to be 0.446 and 0.097 for 3 min. enpoint and 1 hour endpoint, respectively. The Mean % tissue viability compared to negative control (n=3) of the test chemical was determined to be 15.7 % and 3.4 % for 3 min. enpoint and 1 hour endpoint, respectively. Thus, under the experimental condition test chemical was considered to be corrosive to the human skin.

This is supported by the study performed for the test chemical according to OECD 404 (1981), EEC test method B.4, Annex V of EEC Directive 84/449/EEC (September 1984) to evaluate the dermal irritation potential of the test chemical. A paste, (0.5 g test chemical and 0.25 ml water), was applied evenly to 6 cm2 Metalline on a permeable tape (Micropore). This was applied to the right flank of 3 females New Zealand albino rabbits. A control patch without the test substance was applied to the left flank. These were left in place for 4 hours. The test site was cleaned first with a dry tissue and then by swabbed with a dampened tissue. The skin was examined for erythema, eschar formation and oedema at 1, 24, 48 and 72 hours after removal of the patches. Two animals showed slight erythema initially. 1 of these also showed slight edema up to 24 h. These effects were resolved by 48 h. There was no indication of a systemic effect. The modified primary irritation index (PII) of 0.5 was recorded when benzoic acid was applied to the intact rabbit skin under semi-occlusive patch conditions. Based on the scores, the test chemical can be considered to be not irritating to New Zealand albino rabbit skin.

The above results are further supported by the a pre-test for a human maximization study performed to assess the irritation potential of the test chemical.

8% test chemical in petrolatum was applied under occlusion in a closed patch to the backs of 27 male volunteers for 48 hours and the effects were observed. The test chemical was not irritating to the skin of 27 male volunteers after 48 hours exposure.

Hence, test chemical can be considered as not irritating to skin.

Based on the available results for the various test chemicals, the test chemical can be considered to be not irritating to skin on the basis of in-vivo studies. Comparing the above annotations with the criteria of CLP regulation the test chemical can be classified under the category “Not Classified”.

Eye Irritation

The eye irritation potential of the test chemical was assessed based on the available results from the various test chemicals.

The primary eye irritation potential of the test chemicals was evaluated according to Draize method.

3 female Japanese White rabbits (2.0-2.2 kg) were used for the study. The chemical (if solid= 100 mg, if liquid = 0.1ml) was instilled into the conjunctival sac of the left eye.Right eye (untreated eye) served as blank control. The eyes were examined and the grade of ocular lesions was recorded at 1, 24, 48, 72, 96 hours and 7,14,21 days after administration of test chemical. Corneal opacity, erythema, chemosis, secreta and iritis were recorded and classified under Draize method (Draize et.al, 1944).The sum of values, recorded for cornea, conjunctiva and iris was divided by the number of observation times and the average scores (Draize scores) were used as grades of ocular irritation potential of the chemical

The maximum value of Draize score are as follows: Cornea =80, Conjunctiva = 20 and Iris = 5

The average Draize scores after 21 days were as follows: CORNEA = 54.1/80, iris = could not be determined, conjunctiva = 10.3/20.

The test chemical induced severe eye irritation not recovering within 21 days of treatment.

Hence the test chemical can be assessed as irritating to rabbit eyes.             

This is supported by the results of the same study performed for the other test chemical. The average Draize scores after 21 days were as follows: CORNEA = 33.9/80, iris = 0.6, conjunctiva = 5.1.

The test chemical caused moderate eye irritation recovering within 21 days of treatment. Hence, the test chemical can be considered irritating to eyes.

 

The above Draize test results are further supported by another rabbit eye irritation test conducted in 5 healthy albino rabbits to assess the irritation potency of the test chemical. A 0.005 ml aliquot of neat test chemicals was applied to the center of the cornea while the lids were retracted. One minute later the lids were released. The eyes were examined 18–24 h later in strong diffuse daylight and then stained with fluorescein. The test chemical was assigned grade 3 – necrosis since 13–37% of the cornea was visible after fluorescein staining.

Therefore, the test chemical can be considered to be irritating to rabbit eyes.

By applying the weight of evidence approach, the test chemical can be considered to be irritating to eyes. Comparing the annotations with criteria of CLP regulation, the test chemical can be classified under the category "Category 2".

Justification for classification or non-classification

Based on the available results for the various test chemicals, the test chemical can be considered to be not irritating to skin. Comparing the above annotations with the criteria of CLP regulation the test chemical can be classified under the category “Not Classified”. By applying the weight of evidence approach, the test chemical can be considered to be irritating to eyes. Comparing the annotations with criteria of CLP regulation, the test chemical can be classified under the category "Category 2".