2-Butanone, 1,1,1,3,4,4,4-heptafluoro-3-(trifluoromethyl)-

Endpoint summary

Description of key information

In vivo and in vitro skin and eye irritation studies have been conducted on CAS# 756-12-7.

The results of the studies are:

in vivo Eye Irritation (Key): Not irritating when tested according to OECD 405.

in vitro Eye Irritation: Irritating when tested according to OECD 437.

in vivo skin irritation (Key): Not irritating when tested according to OECD 404.

in vitro skin irritation: Not corrosive when tested according to OECD 431.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

2015

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 3M Company, Lot 1
- Expiration date of the lot/batch: 05 June, 2019
- Purity test date: 05 June, 2017

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature
- Stability under test conditions: Stable

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: None, dosed neat

FORM AS APPLIED IN THE TEST: Neat

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River France, L’Arbresle, France
- Age at study initiation: 21-22 weeks old
- Weight at study initiation: 3490-3727 grams
- Housing: On arrival and following assignment to the study, animals were housed individually in labeled cages with perforated floors (Ebeco, Germany, dimensions 67 x 62 x 55 cm) equipped with water bottles. The room(s) in which the animals were kept were documented in the study records. Each cage was clearly labeled.
- Diet (e.g. ad libitum): Pelleted diet for rabbits (Global Diet 2030 Teklad®, Mucedola, Milanese, Italy) was provided once daily throughout the study. In addition, hay (TecniLab-BMI BV, Someren, The Netherlands) was available during the study period. The feed was analyzed by the supplier for nutritional components and environmental contaminants. Results of the analysis were provided by the supplier and are on file at the Test Facility. It is considered that there were no known contaminants in the feed that would interfere with the objectives of the study.
- Water (e.g. ad libitum): Municipal tap-water was freely available to each animal via water bottles. Periodic analysis of the water was performed, and results of these analyses are on file at the Test Facility. It is considered that there were no known contaminants in the water that would interfere with the objectives of the study.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-24
- Humidity (%): 70-73
- Air changes (per hr): At least 10 an hour
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 10 October, 2017 To: 23 October, 2017

Type of coverage:

semiocclusive

Preparation of test site:

other:

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL

VEHICLE : None, dosed neat

NEGATIVE CONTROL : None

POSITIVE CONTROL : None

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: Flank
- % coverage: 2 x 3 cm
- Type of wrap if used: The test article was covered with a metalline patch mounted on micropore tape which was wrapped around the abdomen and secured with Coban elastic bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Yes, with tap water
- Time after start of exposure: 4 hours after application.

OBSERVATION TIME POINTS
Skin reactions were assessed 1, 24, 48 and 72 hours after exposure.

SCORING SYSTEM:
- Method of calculation: Draize

Irritation parameter:

erythema score

Basis:

mean

Time point:

24 h

Score:

0

Max. score:

0

Irritation parameter:

erythema score

Basis:

mean

Time point:

48 h

Score:

0

Max. score:

0

Irritation parameter:

erythema score

Basis:

mean

Time point:

72 h

Score:

0

Max. score:

0

Irritation parameter:

edema score

Basis:

mean

Time point:

24 h

Score:

0.67

Max. score:

1

Reversibility:

fully reversible within: 24 hours

Irritation parameter:

edema score

Basis:

mean

Time point:

48 h

Score:

0

Max. score:

0

Irritation parameter:

edema score

Basis:

mean

Time point:

72 h

Score:

0

Max. score:

0

Irritant / corrosive response data:

Four hours exposure to 0.5 mL of the test article resulted in very slight edema in the treated areas of two of the three rabbits. The skin irritation resolved within 24 hours after exposure in these animals.

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results of the study (PII= 0.22), the test article is not a skin irritant.

Executive summary:

The acute dermal irritation potential of the test article was evaluated in three female rabbits. The study was conducted according to OECD 404 (2015) in compliance with OECD GLP principles. Three female rabbits were exposed to 0.5 mL of the test article by application onto clipped skin. The test article was covered with a metalline patch mounted on micropore tape which was wrapped around the abdomen and secured with Coban elastic bandage. Following a 4 hour exposure to the test article, the site was washed with tap water and skin reactions were assessed at 1, 24, 48 and 72 hours. Four hours exposure to 0.5 mL of the test article resulted in very slight edema in the treated areas of two of the three rabbits. The skin irritation resolved within 24 hours after exposure in these animals. Based on the results of the study (PII= 0.22), the test article is not a skin irritant.

Reference 2

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

19 March, 2012 to 23 March, 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted under GLP conditions

Qualifier:

according to guideline

Guideline:

other: - OECD Guideline no.431:In Vitro Skin Corrosion:Human Skin Model Test (adopted 13 April 2004) - EC No. 440/2008,Part B:Methods for the Determination of Toxicity and other health effects,Guideline B.40 BIS:In Vitro Skin Corrosion:Human Skin Model Test

Deviations:

yes

Remarks:

See 'Principles of method' field.

Principles of method if other than guideline:

1. The absolute mean OD540 (optical density at 540 nm) of the negative control tissues after 1
hour treatment was above the laboratory historical control data range.
Evaluation: An OD540 value above the historical data range represents a high mitochondrial
activity and thus viability as measured with the MTT reaction above the historical control data
range. As the viability of the negative control tissues is high, the study integrity is not adversely
affected by this deviation.

2. The intertissue variability (in viability) of the two tissues after treatment with MTDID 20422 for
3 min was 45% and therefore higher than the acceptability criteria of 30%. Moreover, the
difference between the mean viability of these two tissues and one of the two tissues (%) was
respectively 22% and 29% (acceptance criteria of 15%).
Evaluation: Since the individual viability values of the two tissues treated with MTDID 20422
for 3 min were 95% and 53% respectively (not corrosive) and in addition no corrosive effects
were observed after the 1-hour treatment (individual viability values 97% and 79%), it is
concluded that this deviation does not adversely affect the study.

The study integrity was not adversely affected by the deviations.

GLP compliance:

yes

Species:

other: EpiDerm Skin Model (EPI-200, Lot no.:16288 kit C)

Strain:

other: EpiDerm Skin Model (EPI-200, Lot no.:16288 kit C)

Details on test animals or test system and environmental conditions:

TEST TISSUE
- Source: MatTek Corporation, Ashland MA, U.S.A.
- The model consists of normal, human-derived epidermal keratinocytes which have been cultured to
form a multilayered, highly differentiated model of the human epidermis. It consists of organized basal,
spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid
layers arranged in patterns analogous to those found in vivo. The EpiDerm tissues (surface 0.6 cm²)
were cultured on polycarbonate membranes of 10 mm cell culture inserts.

With exception of the 3-minute and 1-hour exposure of the tissues to the test article and the
concurrent controls, the incubations were carried out in a controlled environment, in which optimal
conditions were a humid atmosphere of 80 - 100% (actual range 89 - 95%), containing 5.0 ± 0.5% CO2
in air in the dark at 37.0 ± 1.0°C (actual range 37.4 - 37.5 °C). Temperature and humidity were
continuously monitored throughout the experiment. The CO2 percentage was monitored once on each working day.

Type of coverage:

other: In vitro system. EpiDerm tissue was treated with the test article.

Preparation of test site:

other: In vitro system.

Vehicle:

unchanged (no vehicle)

Controls:

other: In vitro method. Positive and negative control substances were utilized. Negative control: Milli-Q water (Millipore Corp., Bedford, Mass., USA) Positive control: Potassium hydroxide (KOH; Merck, Darmstadt, Germany), an 8.0 normal solution was prepared.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 50 microliters of the undiluted test substance was added (with pipette) into well plates on top of the skin tissue.

Duration of treatment / exposure:

3 minute exposure (2 of 6 wells), 1 hour exposure (2 of 6 wells).

Details on study design:

TEST SITE
-The test article was added on top of skin tissue in the well (see Figure 1 below)
REMOVAL OF TEST SUBSTANCE
- Washing (if done): Wells were washed with phosphate buffered saline (Invitrogen Corporation, Breda, The Netherlands).
- Time after start of exposure: Washing was done after 3 minute and 1 hour exposure times.
SCORING SYSTEM: Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from 3-(4,5-dimethylthiazaol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) at the end of treatment.

Irritation / corrosion parameter:

other: other: Corr: * relat. mean tissue viab.after 3min. treatm. < 50% * non-corr. after 3min. treatm. is corr. if relat. tissue viab. after 1h treatm.<15%.Non corr: *relat. mean tissue viab. after 3min. treatm. not <50% * relat. tissue viab.after 1h treatm. no

Value:

ca. 0.74 - ca. 0.88

Remarks on result:

other:

Remarks:

Basis: mean. Time point: 3 minute, 1 hour exposure. Reversibility: other: In vitro method.. Remarks: The mean tissue viability of after treatment with the test article was 74% of the negative control after a 3 minute exposure and 88% after 1 hour exposure.. (migrated information)

Other effects / acceptance of results:

Based on the results of the study, the test article is not corrosive in the in vitro skin corrosion test model and met the requirements of the test

Table 3 Mean tissue viability in the in vitro skin corrosion test with MTDID 20422

	3 -minute applicaton viability (percentage of control	1 -hour application viability (percentage of control)
Negative control	100	100
MTDID 20422	74	88
Positive control	13	6

Interpretation of results:

other: not corrosive

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

The mean tissue viability of after treatment with the test article was 74% of the negative control after a 3 minute exposure and 88% after 1 hour exposure. Based on the results of the study, the test article is not corrosive in the in vitro skin corrosion test model and met the requirements of the test

Executive summary:

The skin corrosion potential of the test article (clear colorless liquid) was evaluated in an in vitro human skin model test. This study was performed in compliance with OECD GLP (1997). The study design was based on OECD Guideline No. 431 (04/13/2004) and EC 440/2008 (2008). Human-derived keratinocytes were cultured to form a multilayered, highly differentiated model of the human epidermis. The tissues were kept refrigerated until the day of use, transferred to 6 well plates containing 0.9 mL DMEM medium, and then incubated for 2 hours at 37 C in 5% CO2. The test article (50 uL) was applied to 2 tissues for a 3 minute exposure and to 2 tissues for 1 hour exposure. The negative control (Milli-Q water) and the positive control (Potassium hydroxide) were applied (50 uL of each) in the same manner. Following the exposure periods, the tissues were washed with phosphate buffered saline (PBS). The DMEM medium was replaced with 300 uL MTT medium and then all tissues were again incubated for 3 hours at 37 C in 5% CO2. The tissues were then washed with PBS and formazan was extracted with 2 mL isopropanol. The amount of extracted formazan was determined spectrophotometrically at 540 nm in triplicate. Cell viability was then calculated as a percentage of the mean of the control tissues, and skin corrosion potential calculated from the optical density at 540 nm (OD540) measurements. The mean tissue viability of the tissue after treatment with the test article was 74% of the negative control after a 3 minute exposure and 88% after 1 hour exposure. The controls performed as expected indicating that the test was valid. Based on the results of the study, the test article is not corrosive in the in vitro skin corrosion test model and met the requirements of the test.

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

Study period:

2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

2012

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 3M Company, Lot 1
- Expiration date of the lot/batch: 05 June, 2019
- Purity test date: 05 June, 2017

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature
- Stability under test conditions: Stable

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: None, dosed neat

FORM AS APPLIED IN THE TEST: Neat

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River France, L’Arbresle, France
- Age at study initiation: 12-13 weeks
- Weight at study initiation: 2859-3053 grams
- Housing: On arrival and following assignment to the study, animals were housed individually in labeled cages with perforated floors (Ebeco, Germany, dimensions 67 x 62 x 55 cm) equipped with water bottles. The room(s) in which the animals were kept were documented in the study records. Each cage was clearly labeled.
- Diet (e.g. ad libitum): Pelleted diet for rabbits (Global Diet 2030 Teklad®, Mucedola, Milanese, Italy) was provided once daily throughout the study. In addition, hay (TecniLab-BMI BV, Someren, The Netherlands) was available during the study period. The feed was analyzed by the supplier for nutritional components and environmental contaminants. Results of the analysis were provided by the supplier and are on file at the Test Facility. It is considered that there were no known contaminants in the feed that would interfere with the objectives of the study.
- Water (e.g. ad libitum): Municipal tap-water was freely available to each animal via water bottles. Periodic analysis of the water was performed, and results of these analyses are on file at the Test Facility. It is considered that there were no known contaminants in the water that would interfere with
the objectives of the study.
- Acclimation period: At least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-24 C
- Humidity (%): 40-70
- Air changes (per hr): At least 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 20 November, 2017 To: 06 December, 2017

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL

VEHICLE: None, dosed neat.

Duration of treatment / exposure:

Each animal was treated by instillation of 0.1 mL of the test item, in the conjunctival sac of one of the eyes after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test item.

Observation period (in vivo):

Observations were made 1, 24, 48 and 72 hours after instillation.

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): None
- Time after start of exposure: NA

SCORING SYSTEM: Draize

TOOL USED TO ASSESS SCORE: Fluorescein

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 1 hour

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 24 hours

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24 h

Score:

0

Max. score:

0

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

48 h

Score:

0

Max. score:

0

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

72 h

Score:

0

Max. score:

0

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24 h

Score:

0

Max. score:

0

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

48 h

Score:

0

Max. score:

0

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

72 h

Score:

0

Max. score:

0

Irritation parameter:

iris score

Basis:

mean

Time point:

24 h

Score:

0

Max. score:

0

Irritation parameter:

iris score

Basis:

mean

Time point:

48 h

Score:

0

Max. score:

0

Irritation parameter:

iris score

Basis:

mean

Time point:

72 h

Score:

0

Max. score:

0

Irritation parameter:

other: Conjunctivae Discharge

Basis:

mean

Time point:

other: 1 hour

Score:

0.67

Max. score:

1

Reversibility:

fully reversible within: 24 hours

Irritation parameter:

other: Conjunctivae Discharge

Basis:

mean

Time point:

24 h

Score:

0

Max. score:

0

Irritation parameter:

other: Conjunctivae Discharge

Basis:

mean

Time point:

48 h

Score:

0

Max. score:

0

Irritation parameter:

other: Conjunctivae Discharge

Basis:

mean

Time point:

72 h

Score:

0

Max. score:

0

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24 h

Score:

0

Max. score:

0

Irritation parameter:

chemosis score

Basis:

mean

Time point:

48 h

Score:

0

Max. score:

0

Irritation parameter:

chemosis score

Basis:

mean

Time point:

72 h

Score:

0

Max. score:

0

Irritant / corrosive response data:

Instillation of the test item resulted in irritation of the conjunctivae at the 1 hour examination, which consisted of redness (3/3 animals, scores of 1) and discharge (2/3 animals, scores of 1). The irritation had completely resolved by the 24 hour observation in all animals. No iridial irritation or corneal opacity were observed, and treatment of the eyes with 2% fluorescein 24 hours after test item instillation revealed no corneal epithelial damage. No staining of (peri) ocular tissues by the test item was observed and no test item remnants were seen.

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results of the study, the test article is not an ocular irritant.

Executive summary:

The acute ocular irritation potential of the test article was evaluated in three male rabbits. The study was conducted according to OECD 405 (2012) in compliance with OECD GLP principles. Each animal was treated by instillation of 0.1 mL of the test article, in the conjunctival sac of one of the eyes after gently pulling the lower lid away from the eyeball while the other eye remained untreated to serve as a reference control. The lids were then gently held together for about one second to prevent loss of the test item. Observations were made 1, 24, 48 and 72 hours after instillation with fluorescein being utilized at the 24 hour observation to quantitatively determine corneal epithelial damage. Instillation of the test item resulted in irritation of the conjunctivae at the 1 hour examination, which consisted of redness (3/3 animals, scores of 1) and discharge (2/3 animals, scores of 1). The irritation had completely resolved by the 24 hour observation in all animals. No iridial irritation or corneal opacity were observed, and treatment of the eyes with 2% fluorescein 24 hours after test item instillation revealed no corneal epithelial damage. No staining of (peri) ocular tissues by the test item was observed and no test item remnants were seen. Based on the results of the study, the test article is not an ocular irritant.

Reference 2

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29 March 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted under GLP conditions.

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Deviations:

yes

Remarks:

See 'Principles of method' field.

Qualifier:

according to guideline

Guideline:

EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)

Deviations:

yes

Remarks:

See 'Principles of method' field.

Qualifier:

according to guideline

Guideline:

other: The OTWG of the ICCVAM and the NICEATM, Background Review Document (BRD): current status of in vitro test methods for identifying ocular corrosives and severe irritants: The Bovine Corneal Opacity and Permeability (BCOP) Test Method, March 2006.

Deviations:

yes

Remarks:

See 'Principles of method' field.

Qualifier:

according to guideline

Guideline:

other: In Vitro Techniques in Toxicology Database (INVITTOX) protocol 127. Bovine Opacity and Permeability (BCOP) Assay, 2006.

Deviations:

yes

Remarks:

See 'Principles of method' field.

Qualifier:

according to guideline

Guideline:

other: Gautheron P., Dukic M., Alix D. and Sina J.F., Bovine corneal opacity and permeability test: An in vitro assay of ocular irritancy. Fundam Appl Toxicol 18:442-449, 1992.

Deviations:

yes

Remarks:

See 'Principles of method' field.

Principles of method if other than guideline:

Description of deviations :
1. Additional corneas of the negative and positive control substances were incubated in a horizontal
position for 10  1 minutes at 32  1C.
Evaluation: The historical control data range has been build up by corneas treated at 32  1C.
The comparision of the control values of corneas treated at room temperature and at 32C had no
effect on the results of the study.

2. The corneas treated with the control substances were incubated in a horizontal position for
12 minutes at 32C and for 8 minutes at room temperature.
Evaluation: However since the treatment period was only 1 minute different from the treatment
period mentioned in the protocol and the negative (two out of three corneas at 32C) and positive
control data were within the laboratory historical range, this short deviation of the treatment
period had no effect on the results of the study

3. In the solvent control measurement treated at 32C, the opacity score of one cornea (3) was out
of range after the 10 minutes treatment and this measurement was excluded from the IVIS
determination.
Evaluation: Since the other two measurements (opacity score of 0 for both corneas) were within
the historical control data base (-1 to 1), this deviation in the IVIS determination had no influence
on the study results.

The study integrity was not adversely affected by the deviations.

GLP compliance:

yes

Species:

other: Bovine eyes

Strain:

other: Bovine eyes

Details on test animals or tissues and environmental conditions:

BOVINE CORNEA
- Source: Vitelco's (Hertogenbosch, The Netherlands)
Bovine eyes from young cattle were obtained from the slaughterhouse where the eyes were excised by a slaughterhouse employee as soon as possible after slaughter. Eyes were collected and transported in physiological saline in a suitable container.

Vehicle:

unchanged (no vehicle)

Controls:

other: Control corneas treated with a positive and negative control substances were used.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): Bovine Cornea was immersed in 750 microliters of the test article.

Duration of treatment / exposure:

10 minutes (+/- 1 minute) at room temperature.

Observation period (in vivo):

Opacity and permiability determination occured 120 +/- 10 minutes after treatment. Corneas were incubated in Eagle's Minumum Essentail Medium between treatment and analysis.

Details on study design:

Negative control substance: Physiological saline (Merck, Darmstadt, Germany)
Positive control: 10% (w/v) Benzalkonium Chloride (Sigma-Aldrich Chemie GmbH, Germany) solution prepared in physiological saline.

Irritation parameter:

in vitro irritation score

Run / experiment:

Mean

Value:

29.4

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

An IVIS score of 29.4 indicated that the test article is not a severe irritant or corrosive in the Bovine Corneal Opacity and Permeability test.

Interpretation of results:

moderately irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Based on the results of the test, the test article is moderate irritant to the cornea (IVIS = 29.4).

Executive summary:

The corneal irritation and damage potential of the test article (clear and colorless liquid, purity 99.99%) was tested in the Bovine Corneal Opacity and Permeability test (BCOP test). The study was performed in compliance with OECD GLP (98)17 (1997). The test method was based on OECD no. 437 (2009), EC No. 440/2008 B. 47 (2010), OTWG- ICCVAM-NICEATM (2006), INVITTOX 127 (2006), and Gautheron, P. et al. 18: 442-449 (1992). Corneas were prepared in cell culture medium and incubated at 32 C for at least 1 hour prior to exposure. Corneas (3/group) were treated with 0.75 mL of the undiluted test material and incubated in a horizontal position for 10 minutes at room temperature. A positive control (10% w/v Benzalkonium Chloride) and a negative control (physiological saline) were tested in parallel with the test material. At the end of the of the exposure period, the corneas were rinsed and incubated in fresh cell culture medium for 120 +/- 10 minutes at 32 +/- 1 C. Opacity was evaluated using an opacitometer after the 2 hour post-exposure incubation. Following opacity readings, the cell culture medium was replaced with Na-fluorescein medium and incubated for approximately 90 minutes. Following the 90 minute exposure, permeability was measured. The mean in vitro irritation score (IVIS) was calculated and the scores were classified according to protocol defined categories. For the test article, the IVIS = 29.4 after 10 minutes. The mean opacity score was 4 (range 3-4) and the mean permeability score was 1.694 (range1.009 – 2.466). Controls performed as expected. A pH effect of the test substance was observed on the rinsing medium, and the corneas were rinsed until no color change of the medium was observed. Based on the results of the test, the test article is moderate irritant to the cornea (IVIS = 29.4).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Justification for classification or non-classification

CAS# 756-12-7 is not irritating to Eye and Skin according to CLP classification criteria.