1,4-bis(p-tolylamino)anthraquinone

Endpoint summary

Description of key information

Skin Irritation

Skin irritation study was conducted on New Zealand White rabbits under occlusive condition to assess the irritation potential of the test chemical. The study was performed according to OECD guideline 404. 

No destructions or irreversible alterations of the treated skin and no corrosion effects had occurred on the skin.

Also the mean primary irritation index was estimated to be 0.0.

Thus on basis of scores and observed effects, the test chemical was considered to be not irritating to the rabbits skin under tested experimental condition.

Eye Irritation

An ocular irritation study of test chemical was conducted on New Zealand White rabbits according to OECD guideline 405.  

The test chemical showed no irritation when applied on the rabbit eye mucosa. No discoloration of the cornea and conjunctivae was observed in the rabbits during the entire test period which could be related to compound effects. No corrosion effect was observed at each of the measuring intervals.

Hence, the test chemical was considered to be not irritating to eyes.

The eye irritation potential of the test chemical was also evaluated in an Eye irritation test with EpiOcular™ (MatTek ,MA) was conducted according to OECD 492.

The mean %tissue viability was 100%, since the treated tissues had viability greater than 60%, the test substance was classified as non-irritating. Hence, the test chemical was considered not to be an ocular irritant and it can be classified under the category “Not Classified”.

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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:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Principles of method if other than guideline:

Skin irritation study of test chemical was conducted on New Zealand White rabbits under occlusive condition according to OECD guideline 404.

GLP compliance:

no

Remarks:

pre-dates GLP regulation

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: KLEENTIERFARM MADOERIN AG; 4414 FUELLINSDORF; SWITZERLAND
- Age at study initiation: 12 -16 weeks
- Weight at study initiation: 2.3 to 3.0 kg
- Housing: The animals were caged individually in stainless steel cages.
- Diet (e.g. ad libitum): Standard kliba rabbit maintenance food;ad libitum.
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 4

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 2 degC
- Humidity (%): 55 +/- 10 %
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12 hours/day

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

other: Polyethylene glycol / Saline (70/30 PARTS)

Controls:

not specified

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 ml
- Concentration : 50%

VEHICLE
- Amount(s) applied (volume or weight with unit): no data
- Concentration (if solution): 70% polyethylene glycol + 30 % saline

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

3 (2 males and 1 female)

Details on study design:

TEST SITE
- Area of exposure: 6,25 cm²
- Type of wrap if used: occlusive

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no data
- Time after start of exposure: 4 hours

OBSERVATION TIME POINTS
(indicate if minutes, hours or days): Observed at 1, 24, 48 and 72 hours.

SCORING SYSTEM:
- Method of calculation: Draize method.

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Time point:

24/48/72 h

Score:

0

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

The test chemical showed no irritation,when applied to intact rabbit skin.

No destructions or irreversible alterations of the treated skin were observed. Thus it was concluded that no corrosion effect had occured on the skin.

Other effects:

In the area of application a moderate to slight greyish discoloration of the skin was observed. This effect was recorded in the rabbits during the entire observation period.

Interpretation of results:

other: not irritating

Conclusions:

In the area of application a moderate to slight greyish discoloration of the skin was observed. This effect was recorded in the rabbits during the entire observation period.
No destructions or irreversible alterations of the treated skin and no corrosion effects had occurred on the skin.
Also the mean primary irritation index was estimated to be 0.0.
Thus on basis of scores and observed effects, the test chemical was considered to be not irritating to the rabbits’ skin under tested experimental condition.

Executive summary:

Skin irritation study was conducted on New Zealand White rabbits under occlusive condition to assess the irritation potential of the test chemical. The study was performed according to OECD guideline 404. 

The test chemical was applied to the intact skin of three rabbits (2 male and 1 female) for exposure period of 4 hours under occlusive condition at a dose of 50% in Polyethylene glycol / Saline (70/30 Parts). The skin reactions were recorded at 1, 24, 48 and 72 hours according to Draize method.

In the area of application a moderate to slight greyish discoloration of the skin was observed. This effect was recorded in the rabbits during the entire observation period.

No destructions or irreversible alterations of the treated skin and no corrosion effects had occurred on the skin.

Also the mean primary irritation index was estimated to be 0.0.

Thus on basis of scores and observed effects, the test chemical was considered to be not irritating to the rabbits skin under tested experimental condition.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

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 report

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Principles of method if other than guideline:

Evaluation of occular irritation produced by the test chemical introduced into the conjuntival sac of the rabbit eye.

GLP compliance:

no

Remarks:

pre-dates GLP regulation

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: KLEINTIERFARM MADOERIN AG; 4414 FUELLINSDORF / SWITZERLAND
- Age at study initiation: 12 weeks
- Weight at study initiation: 2,3 -3,0 kg
- Housing: The animals were caged individually in stainless steel cages.
- Identification: Individually by numbered ear tags and cage numbers.
- Diet (e.g. ad libitum): Pelleted standard KLIBA 341, rabbit maintenance diet,ad libitum.
- Water (e.g. ad libitum): Tap water,ad libitum.
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+/- 2 degC
- Humidity (%): 55 +/- 10 %
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12 hours/day

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 100 mg/rabbit eye.
- Concentration (if solution): pure

VEHICLE
- none

Duration of treatment / exposure:

single application

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3 (2 male and 1 female)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Eyes were not washed.

SCORING SYSTEM: Draize method

TOOL USED TO ASSESS SCORE: No data

Irritation parameter:

other: Primary irritation index

Basis:

mean

Time point:

24/48/72 h

Score:

1.6

Reversibility:

not specified

Remarks on result:

no indication of irritation

Remarks:

Unrinsed Eyes

Irritant / corrosive response data:

The test chemical showed no irritation when applied on the rabbit eye mucosa.
No discoloration of the cornea and conjunctivae was observed in the rabbits during the entire test period which could be related to compound effects.
No corrosion effect was observed at each of the measuring intervals.

Other effects:

No acute toxicological signs were observed in the animals during the test period.

Interpretation of results:

other: not irritating

Conclusions:

The test chemical showed no irritation when applied on the rabbit eye mucosa. No discoloration of the cornea and conjunctivae was observed in the rabbits during the entire test period which could be related to compound effects. No corrosion effect was observed at each of the measuring intervals.
Hence, the test chemical was considered to be not irritating to eyes.

Executive summary:

An ocular irritation study of test chemical was conducted on New Zealand White rabbits according to OECD guideline 405.  

About 100mg of undiluted chemical was instilled into the one eye of each rabbit whereas the other untreated eye served as control.

The treated eyes were not rinsed and the ocular lesions were scored after 72 hours according to Draize method.

The test chemical showed no irritation when applied on the rabbit eye mucosa. No discoloration of the cornea and conjunctivae was observed in the rabbits during the entire test period which could be related to compound effects. No corrosion effect was observed at each of the measuring intervals.

Hence, the test chemical was considered to be not irritating to eyes.

Reference 2

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

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 journals

Qualifier:

according to guideline

Guideline:

OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)

Principles of method if other than guideline:

The eye irritation potential of the test chemical was evaluated in an Eye irritation test with EpiOcular™ (MatTek , MA) was conducted according to OECD 492

GLP compliance:

not specified

Species:

other: EpiOcular™ (MatTek, USA)

Vehicle:

other: Dulbecco’s Phosphate Buffered Saline (D-PBS)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

50 mg of solid material

Duration of treatment / exposure:

360 min

Duration of post- treatment incubation (in vitro):

Tissues were then transferred to a six-well plate containing 1 mL of warm assay medium (37 ℃) and incubated for 18 hours

Number of animals or in vitro replicates:

Duplicates

Details on study design:

Pre-treatment of the tissues:
Tissues were transferred to a six-well plate containing 1.0 mL of pre-warmed assay medium. Tissues were then incubated overnight at 37 ℃ in a humidified atmosphere of 5% CO2. After incubation,
tissues were pre-treated with 20 mL of Dulbecco’s Phosphate Buffered Saline (D-PBS) and incubated for 30 min, prior to exposure to the test chemical.

Study Design:
The experimental steps after pre- treatment of solid materials, was as follows:
1) Treatment, 50 mg of solid material for 360 min;
2) Rinse, for rinsing the tissues, three clean beakers, each containing 100 mL of Dulbecco’s Phosphate Buffered Saline, were prepared; two tissues that were treated with the same chemical were washed at a time; and each beaker was rinsed three times;
3) Post-soak, after the final rinse, tissues were immersed in 5 mL of assay medium in a 12-well plate for 25 min for solid materials at room temperature;
4) Post-incubation, tissues were then transferred to a six-well plate containing 1 mL of warm assay medium (37 ℃) and incubated for 18 hrs for solid materials.
Then, through MTT assay, the viability of tissues was measured to determine the eye irritation potential of test materials.

Description of the method used to quantify MTT formazan:
Because colorants can interfere with MTT assay, additional color interference tests were conducted for EpiOcularTM according to TG 492. First, the light absorbance of colorants in water and isopropanol were examined for interference at 570 nm, which is in the same range with MTT for formazan, revealing significant color interferences. To consider the interference, an additional control, a non-specific color in living tissue (NSCliving) control, which underwent the entire testing procedure, although it was incubated with the medium instead of MTT solution during the MTT incubation step.
True tissue viability was calculated as [%Viability test] - [%NSCliving], and results showed similar tissue viabilities with those obtained without NSCliving,

Statistics:
The data are expressed as mean ± standard deviation of the mean of duplicates to analyze statistical differences from the control group. For RhCE, data are presented as the mean ± SD or mean ± 1/2 difference between duplicate wells.

Irritation parameter:

other: % tissue viability

Run / experiment:

1

Value:

100

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

The mean %tissue viability was 100%, since the treated tissues had viability greater than 60%, the test substance was classified as non-irritating.

Decisions on eye irritancy of the coal-tar dyes according to UN GHS in in vitro eye irritation tests

CAS	EpiOcular^TM (≤60%, Irritant)	MCTT HCE^TM (≤45%, Irritant)	Bottom –up approach		Reference & ECHA
			EpiOcular^TM (≤60%, Irritant)	MCTT HCE^TM (≤45%, Irritant)
128 -80 -3	NC	NC	NC	NC	NC,CAT 2

 

Interpretation of results:

other: Not irritating

Conclusions:

The mean %tissue viability was 100%, since the treated tissues had viability greater than 60%, the test substance was classified as non-irritating. Hence, the test chemical was considered not to be an ocular irritant and it can be classified under the category “Not Classified”.

Executive summary:

The eye irritation potential of the test chemical was evaluated in an Eye irritation test with EpiOcular™ (MatTek ,MA) was conducted according to OECD 492.

EpiOcular™ (MatTek, USA)tissues were transferred to a six-well plate containing 1.0 mL of pre-warmed assay medium.Tissues were then incubated overnight at 37℃in a humidified atmosphere of 5% CO2. After incubation,tissues were pre-treated with 20 mL of Dulbecco’s Phosphate Buffered Saline (D-PBS) and incubated for 30 min, prior to exposure to the test chemical. The experimental steps after pre- treatment of solid materials, was as follows:

1) Treatment, 50 mg of solid material for 360 min;

 2) Rinse, for rinsing the tissues, three clean beakers, each containing 100 mL of Dulbecco’s Phosphate Buffered Saline, were prepared; two tissues that were treated with the same chemical were washed at a time; and each beaker was rinsed threetimes;

 3) Post-soak, after the final rinse, tissues were immersed in 5 mL of assay medium in a 12-well plate for 25 min for solid materials at room temperature;

4) Post-incubation, tissues were then transferred to a six-well plate containing 1 mL of warm assay medium (37℃) and incubated for 18 hrsfor solid materials.

Then, through MTT assay, the viability of tissues was measured to determine the eye irritation potential of test materials. Because colorants can interfere with MTT assay, additional color interference tests were conducted for EpiOcularTM according to TG 492. First, the light absorbance of colorants in water and isopropanol were examined for interference at 570 nm, which is in the same range with MTT for formazan, revealing significant color interferences.To consider the interference, anadditional control, a non-specific color in living tissue (NSCliving) control, which underwent the entire testing procedure, although it was incubated with the medium instead of MTT solution during the MTT incubation step.

True tissue viability was calculated as [%Viability test] - [%NSCliving], and results showed similar tissue viabilities with those obtained without NSCliving.The treated tissues had viability greater than 60%, the test substance was classified as non-irritating, and if less than 60%, the test substance was classified as irritating.

The mean %tissue viability was 100%, since the treated tissues had viability greater than 60%, the test substance was classified as non-irritating.Hence, the test chemical was considered not to be an ocular irritant and it can be classified under the category “Not Classified”.

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 in living organisms. These include in vivo experimental studies on rabbits for the test chemical. The results are summarized below:

Skin irritation study was conducted on New Zealand White rabbits under occlusive condition to assess the irritation potential of the test chemical. The study was performed according to OECD guideline 404. 

The test chemical was applied to the intact skin of three rabbits (2 male and 1 female) for exposure period of 4 hours under occlusive condition at a dose of 50% in Polyethylene glycol / Saline (70/30 Parts). The skin reactions were recorded at 1, 24, 48 and 72 hours according to Draize method.

In the area of application a moderate to slight greyish discoloration of the skin was observed. This effect was recorded in the rabbits during the entire observation period.

No destructions or irreversible alterations of the treated skin and no corrosion effects had occurred on the skin.

Also the mean primary irritation index was estimated to be 0.0.

Thus on basis of scores and observed effects, the test chemical was considered to be not irritating to the rabbits skin under tested experimental condition.

This is supported by the results of a primary dermal irritation study conducted on intact skin of the inner surface of the ear of two adult New Zealand White rabbits under occlusive conditions to assess the irritation potential of the test chemical.

The rabbits were exposed to 500 mg of undiluted test substance at the inner side of the ear flap of one ear secured by an adhesive bandage for 24 hours. The contralateral ear served as control.

Animals were then observed for erythema and edema upto7 days.

The skin reactions were was scored by the method of (US-Departments of Agriculture, Federal Register Vol.38, No.187, p. 27019, 1973 resp. DRAIZE, J.H., Appraisal of the safety of chemicals in foods,drugs and cosmetics, Association of Food and Drug Officials of the US, p.46, 1959).

Since the chemical did not induce erythema or edema in both the rabbits, the test chemical was considered to be not irritating on intact skin of the inner surface of the ear of two adult New Zealand White rabbits under the tested experimental conditions.

The above results are further supported by another skin irritation study carried out in New Zealand white rabbit to assess the irritation potential of the test chemical.

A measurement of 0.5 g of the test material was applied to a 1 in. x 1in. gauze pad, moistened with saline, and secured with surgical tape to two abraded and two unabraded sites in 6 New Zealand White rabbits. The trunk of the rabbit was covered first with plastic wrap and then with stockinette to prevent disturbance of the test sites.

Application of 0.5 g of the test chemical to 2 abraded and 2 unabraded test sites of 6 rabbits for 24 hours produced barely perceptible erythema to one abraded test site of one male and two female rabbits, and one unabraded site of one male rabbit. This erythema resolved in all cases by 72 hours. No edema was observed at any of the sites during the 24 and 72 hour observation period. The Primary Irritation Score for this test sample was 0.08. Clinical signs indicative of systemic toxicity were not observed. Hence the test chemical was considered to be not irritating to skin.

The results of the in vivo experimental studies are in mutual agreement with each other indicating a very strong possibility that the test chemical is indeed not irritating to skin. Comparing the above annotations with the criteria of the CLP regulation the test chemical can be classified under the category “Not Classified”.

Eye Irritation

Various studies have been reviewed to determine the extent of ocular damage caused by the test chemical to living organisms. These include in vivo experiments on rabbits as well as in vitro experimental results for the test chemical. The studies are summarized below:

An ocular irritation study of test chemical was conducted on New Zealand White rabbits according to OECD guideline 405.  

About 100mg of undiluted chemical was instilled into the one eye of each rabbit whereas the other untreated eye served as control.

The treated eyes were not rinsed and the ocular lesions were scored after 72 hours according to Draize method.

The test chemical showed no irritation when applied on the rabbit eye mucosa. No discoloration of the cornea and conjunctivae was observed in the rabbits during the entire test period which could be related to compound effects. No corrosion effect was observed at each of the measuring intervals.

Hence, the test chemical was considered to be not irritating to eyes.

The above study is supported another OECD 405 Guideline study conducted on New Zealand White rabbits to determine the ocular irritation potential of the test chemical.  

About 100mg of undiluted chemical was instilled into the one eye of each rabbit whereas the other untreated eye served as control.

The treated eyes were not rinsed and the ocular lesions were scored after 72 hours according to Draize method.

The test chemical showed no irritation when applied on the rabbit eye mucosa. No discoloration of the cornea and conjunctivae was observed in the rabbits during the entire test period which could be related to compound effects. No corrosion effect was observed at each of the measuring intervals.

Hence, the test chemical was considered to be not irritating to eyes.

These results are supported by an eye irritation study carried out on New Zealand white rabbits to assess the irritation potential of the test chemical. 100 mg of the test sample was instilled into the everted lower lid of the right eye. The left eye remained untreated and served as control. The treated eyes remained unwashed and were observed for corneal opacity, iritis, hyperemia, chemosis or discharge till 72 hours. Clinical signs indicative of systemic toxicity were not observed.

The test chemical failed to produce corneal opacity, iritis, hyperemia, chemosis or discharge throughout the 72 hour observation period. The test chemical was considered to be non-irritating to unirrigated eyes of rabbits.

These results are further supported by another study performed on rabbits to assess the ocular irritation potential of the test chemical was assessed in rabbits. 20 mg of the test chemical was instilled into the eyes of rabbits and observed for 24 hours for signs of irritation.

Moderate irritation effect was observed after 24 hours exposure to the test chemical. Hence, the test chemical was considered to be irritating to eyes.

The eye irritation potential of the test chemical was also evaluated in an Eye irritation test with EpiOcular™ (MatTek ,MA) was conducted according to OECD 492.

EpiOcular™ (MatTek, USA)tissues were transferred to a six-well plate containing 1.0 mL of pre-warmed assay medium.Tissues were then incubated overnight at 37℃in a humidified atmosphere of 5% CO2. After incubation,tissues were pre-treated with 20 mL of Dulbecco’s Phosphate Buffered Saline (D-PBS) and incubated for 30 min, prior to exposure to the test chemical. The experimental steps after pre- treatment of solid materials, was as follows:

1) Treatment, 50 mg of solid material for 360 min;

 2) Rinse, for rinsing the tissues, three clean beakers, each containing 100 mL of Dulbecco’s Phosphate Buffered Saline, were prepared; two tissues that were treated with the same chemical were washed at a time; and each beaker was rinsed threetimes;

 3) Post-soak, after the final rinse, tissues were immersed in 5 mL of assay medium in a 12-well plate for 25 min for solid materials at room temperature;

4) Post-incubation, tissues were then transferred to a six-well plate containing 1 mL of warm assay medium (37℃) and incubated for 18 hrsfor solid materials.

Then, through MTT assay, the viability of tissues was measured to determine the eye irritation potential of test materials. Because colorants can interfere with MTT assay, additional color interference tests were conducted for EpiOcularTM according to TG 492. First, the light absorbance of colorants in water and isopropanol were examined for interference at 570 nm, which is in the same range with MTT for formazan, revealing significant color interferences.To consider the interference, anadditional control, a non-specific color in living tissue (NSCliving) control, which underwent the entire testing procedure, although it was incubated with the medium instead of MTT solution during the MTT incubation step.

True tissue viability was calculated as [%Viability test] - [%NSCliving], and results showed similar tissue viabilities with those obtained without NSCliving.The treated tissues had viability greater than 60%, the test substance was classified as non-irritating, and if less than 60%, the test substance was classified as irritating.

The mean %tissue viability was 100%, since the treated tissues had viability greater than 60%, the test substance was classified as non-irritating. Hence, the test chemical was considered not to be an ocular irritant and it can be classified under the category “Not Classified”.

The above in vitro result is supported by an Eye irritation test with MCTT HCE™ assay to determine the irritation parameter of the test chemical. MCTT HCETMwas prepared from primary human limbal cells. After receiving the tissues, 900 mL of fresh maintenance medium was replenished and incubated at 37°C, 5% CO2 for 24 h. The experimental steps after pre-

treatment of solid materials, was as follows:

Test material solutions at various concentrations were prepared by serial dilution in PBS and were treated at50 mg for solids (100% coal-tar dyes) per well, after which MCTT HCETMwas incubated for 60 minutes. After the exposure, the models were washed four times with PBS and further incubated with 900 ml of a new maintenance medium for 16 hrs (post-incubation period) at 37°C, 5% CO2.For the measurement of tissue viability, tissues were incubated with 300 μL of WST-1 (4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate) and optical density was measured at 450 nm. For the measurement of tissue viability, tissues were incubated with 300 μL of WST-1 (4-[3-(4-iodophenyl)-2-(4 -nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate) and optical density was measured at 450 nm.

Negative control, PBS, and coal-tar dyes showing cell viability over 45%, were determined as non-irritant materials. Positive control (2% of SLS) and coal-tar dyes that reduced cell viability below 45% were determined to be irritant materials.

For histological evaluation, the MCTT HCETM was fixed in 10% neutral buffered formalin to process it for embedding in paraffin. Five-mm vertical sections were cut and stained with hematoxylin and eosin for examination by Olympus BX43 microscope.

The mean %tissue viability was 100%, since the treated tissues showed cell viability over 45%, the test chemical was determined to be non-irritant materials.

Hence, the test chemical was considered to be not an ocular irritant and it can be classified under the category “Not Classified”.

Eventhough the results of one in vivo study indicates a possibility that the test chemical can cause moderate irritation to eyes, but the remaining in vivo and in vitro studies suggest other wise. Also the results of the Invivo and in vitro Guideline studies are in mutual agreement with each other, indicating a very strong possibility of the test chemical being not irritating to eyes. Comparing the above annotations with the criteria of the CLP regulation the test chemical can be classified under the category “Not Classified”.

Justification for classification or non-classification

The results of the in vivo experimental studies are in mutual agreement with each other indicating a very strong possibility that the test chemical is indeed not irritating to skin.

This is also supported by the weakly acidic nature of the substance (pH= 5.71). Comparing the above annotations with the criteria of the CLP regulation the test chemical can be classified under the category “Not Classified”.

Eventhough the results of one in vivo study indicates a possibility that the test chemical can cause moderate irritation to eyes, but the remaining in vivo and in vitro studies suggest other wise. Also the results of the Invivo and in vitro Guideline studies are in mutual agreement with each other, indicating a very strong possibility of the test chemical being not irritating to eyes. This is also supported by the weakly acidic nature of the substance (pH= 5.71). Comparing the above annotations with the criteria of the CLP regulation the test chemical can be classified under the category “Not Classified”.