Reaction mass of (2E)-Tridec-2-enenitrile and (2Z)-Tridec-2-enenitrile and (3E)-Tridec-3-enenitrile and (3Z)-Tridec-3-enenitrile

Endpoint summary

Description of key information

Skin:
The test substance was shown to be not irritating to skin.
Eyes:
The test substance was shown to be not irritating to the eyes.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2015-06-03 to 2015-07-20

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Version / remarks:

2013

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Version / remarks:

2009

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Justification for test system used:

In an international prevalidation study performed by ECVAM, the in vitro skin irritation test using the human skin model EpiDerm™ and EpiSkin™ and measurement of cell viability by dehydrogenase conversion of MTT into a blue formazan salt have turned out as a sufficiently promising predictor for skin irritancy potential.

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: Epi-200- SIT Kit
- Tissue batch number: Lot number: 21680
- Arrival date: 2015-07-14
- Date of initiation of testing: 2015-07-14 (pre-incubation)

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 35 minutes at 37 ± 1.5 °C, 25 min at room temperature
- Temperature of post-treatment incubation: 37 ± 1.5 °C

REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps: at least 15 times, volume not provided
- Observable damage in the tissue due to washing: no data provided
- Modifications to validated SOP: no

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL
- Incubation time: 3 h
- Spectrophotometer: Versamax® Molecular Devices, Softmax Pro, version 4.7.1
- Wavelength: 570 nm
- Filter: 570 ± 1 nm

NUMBER OF REPLICATE TISSUES: 3

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 1

PREDICTION MODEL / DECISION CRITERIA
- The test substance is considered to be irritant to skin if the mean tissue viability is ≤ 50 %.
- The test substance is considered to be non-irritant to skin if the mean tissue viability is > 50 %

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount applied: due to purity adjustment 30.4 μL (47 μL/cm2)
- Concentration: unchanged

NEGATIVE CONTROL
- Amount applied: 30 μL DPBS

POSITIVE CONTROL
- Amount applied: 30 μL
- Concentration: 5 % SLS solution in deionised water

Duration of treatment / exposure:

60 minutes

Duration of post-treatment incubation (if applicable):

post-treatment: 42 h
MTT: 3 h treatment and 70 h extraction

Number of replicates:

3

Irritation / corrosion parameter:

% tissue viability

Value:

90.6

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

100.0 %

Positive controls validity:

valid

Remarks:

6.8 %

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: No.
- Direct-MTT reduction: Optical evaluation of the MTT-reducing capacity of the test item after 1 hour incubation with MTT-reagent did not show blue colour. The test substance is no MTT reducer.
- Colour interference with MTT: The optical pre-experiment (colour interference pre-experiment) to investigate the test item’s colour change potential in water did not led to a change in colour. The test substance does not interfer with the color.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes. After treatment with the negative control the absorbance values were well within the required acceptability criterion of mean OD ≥ 0.8 and ≤ 2.8 for the 60 minutes treatment interval thus showing the quality of the tissues.
- Acceptance criteria met for positive control: Yes. Treatment with the positive control induced a decrease in the relative absorbance as compared to the negative control to 6.8% thus ensuring the validity of the test system.
- Acceptance criteria met for variability between replicate measurements: Yes. The relative standard deviations between the % variabilities of the test item, the positive and negative controls in the main test were below 16 %, thus ensuring the validity of the study.

Dose Group	Exposure Interval	Absorbance 570 nm Tissue 1*	Absorbance 570 nm Tissue 2*	Absorbance 570 nm Tissue 3*	Mean Absorbance of 3 Tissues	Rel. Absorbance [%] Tissue 1, 2 + 3**	Relative Standard Deviation [%]	Mean rel. Absorbance [% of Negative Control]***
Negative control	1 h	1.751	1.579	1.844	1.724	101.5 91.5 106.9	7.8	100.0
Positive control	1 h	0.129	0.115	0.109	0.118	7.5 6.7 6.3	8.6	6.8
Test item	1 h	1.862	1.860	1.923	1.881	85.5 79.6 106.8	15.8	90.6

* Mean of three replicate wells after blank correction

** relative absorbance [rounded values]: (100x(absorbance tissue)/ mean absorbance negative control

*** relative absorbance [rounded values]: (100x(mean absorbance test item/positive control))/ mean absorbance negative control

 

The mean relative absorbance value of the test item, corresponding to the cell viability,

decreased to 90.6% (threshold for irritancy:≤50%), consequently the test item was not irritant to skin.

Interpretation of results:

GHS criteria not met

Conclusions:

The test substance was determined not to be irritating in the in vitro human skin model test with EpiDerm.

Executive summary:

An in vitro study was performed to assess the irritation potential of the test item by means of the Human Skin Model Test according to OECD 439. The test item did not reduce MTT (test for direct MTT reduction), and it did not change colour when mixed with deionised water (test for colour interference). Also its intrinsic colour was not intensive. Consequently, additional tests with freeze-killed or viable tissues were not necessary. Each three tissues of the human skin model EpiDerm were treated with the test item, the negative or the positive control for 60 minutes. 30.4 μL of the test item were applied to each tissue, and spread to match the surface of the tissue. 30 μL of either the negative control (DPBS) or the positive control (5% SLS) were applied to each tissue. After treatment with the negative control the absorbance values were well in the required range of the acceptability criterion of mean OD ≥ 0.8 and ≤ 2.8 for the 60 minutes treatment interval, thus assuring the quality of the tissues. Treatment with the positive control induced a sufficient decrease in the relative absorbance as compared to the negative control for the 60 minutes treatment interval, and thus assuring the validity of the test system. After treatment with the test item the mean relative absorbance value decreased to 90.6 % compared to the relative absorbance value of the negative control. This value is above the threshold for irritancy of ≤ 50 %. Therefore, the test item is not considered to possess an irritant potential. In conclusion, it can be stated that in this study and under the experimental conditions reported, the substance is not irritating to skin.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2015-05-29

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2013

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
- Source: abattoir
- Number of corneae: 9
- Characteristics of donor animals: 9 month old
- Storage, temperature and transport conditions of ocular tissue: The isolated eyes were transported to the laboratory in HBSS at ambient temperature. The corneae were isolated on the same day after delivery of the eyes.
- Time interval prior to initiating testing: The corneae were directly used in the BCOP test on the same day.
- indication of any existing defects or lesions in ocular tissue samples: Eyes presenting defects such as vascularization, pigmentation, opacity and scratches were discarded.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.75 mL

Duration of treatment / exposure:

10 min

Duration of post- treatment incubation (in vitro):

After the test item or control items were rinsed off from the application side with saline, the corneae were incubated at 32 ± 1 °C for further two hours in a vertical position, followed by a second opacity reading.

Number of animals or in vitro replicates:

3 corneas were used per group (test item, negative and positive control)

Details on study design:

SELECTION AND PREPARATION OF CORNEAS
All eyes were carefully examined macroscopically for defects. Those presenting defects such as vascularization, pigmentation, opacity and scratches were discarded. The cornea was carefully removed from the eye using scalpel and rounded scissors. A rim of about 2 mm of tissue (sclera) was left for stability and handling of the isolated cornea. Each isolated cornea was mounted in a specially designed cornea holder that consists of anterior and posterior compartments, which interlace with the epithelial and endothelial sides of the cornea.

QUALITY CHECK OF THE ISOLATED CORNEAS
The basal opacity of all corneae was recorded. Each corneae with a value of the basal opacity > 7 was discarded.

NUMBER OF REPLICATES
Sets of three corneae were used for treatment with the test item and the negative and positive controls.

NEGATIVE CONTROL USED
Saline (0.9% NaCl in deionised water)

POSITIVE CONTROL USED
2-Ethoxyethanol (purity: 99%)

APPLICATION DOSE AND EXPOSURE TIME
0.75 mL of undiluted substance, 10 min exposure time

POST-INCUBATION PERIOD: yes, two hours after rinsing

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: no number determined. The test item was rinsed off.

METHODS FOR MEASURED ENDPOINTS
- Corneal opacity: Changes in the light transmission passing through the corneae were determined using an opacitometer.
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of spectrophotometry (OD490)

SCORING SYSTEM: In Vitro Irritancy Score (IVIS) was used.

DECISION CRITERIA:
IVIS ≤ 3: No Category (according to GHS)
IVIS >3 ≤ 55: No prediction can be made
IVIS > 55: Serious eye damage according to CLP/EPA/GHS (Cat 1)

Irritation parameter:

in vitro irritation score

Run / experiment:

Mean

Value:

0.5

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

0.69

Positive controls validity:

valid

Remarks:

79.37

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: Not reported.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes. With the negative control (saline) neither an increase of opacity nor permeability of the corneae could be observed (mean IVIS = 0.69).
- Acceptance criteria met for positive control: Yes. The positive control (2-Ethoxyethanol) was tested undiluted and showed clear opacity and distinctive permeability of the corneae (mean IVIS = 79.37)
- Range of historical values if different from the ones specified in the test guideline: Negative control: 0.87 – 1.42; positive control: 56.25 – 80.65

Results after 10 min incubation time

Test group	Opacity value = Difference (t130-t0) of Opacity		Permeability at 490 nm (OD490)		IVIS	Mean IVIS	Proposed in vitro Irritancy Score
		Mean		Mean
Negative control	-1	-0.33	0.082	0.068	0.23	0.69	Not categorized
	0		0.065		0.98
	0		0.058		0.87
Positive control	50.33		1.348		70.55	79.37	Category 1
	67.33		1.395		88.25
	53.33		1.731		79.29
Test item	0.33		-0.008		0.21	0.50	Not categorized
	0.33		-0.013		0.13
	1.33		-0.011		1.16

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results of this in vitro assay, the test substance is considered as not requiring classification for eye irritation or serious eye damage.

Executive summary:

The in vitro study was performed to assess the corneal damage potential of the test substance by means of the BCOP assay using fresh bovine corneae. The study was conducted according to OECD 437. After a first opacity measurement of the fresh bovine corneae (t0), the neat test item, the positive, and the negative controls were applied to corneae fixed in an incubation chamber in horizontal position for 10 minutes at 32 ± 1 °C. The posterior chamber contained incubation medium. After the incubation phase the test item, the positive, and the negative controls were each rinsed from the corneae. Further, the corneae were incubated for another 120 minutes at 32 ± 1 °C in a vertical position, while the anterior chamber contain incubation medium as well. Afterwards, opacity was measured a second time (t130). After the opacity measurements permeability of the corneae was determined by measuring spectrophotometrically the transfer of sodium fluorescein after incubation in a horizontal position for 90 minutes at 32 ± 1 °C. With the negative control (0.9 % (w/v) NaCl solution in deionised water) neither an increase of opacity nor permeability of the corneae could be observed. The positive control (2-Ethoxyethanol) showed clear opacity and distinctive permeability of the corneae corresponding to a classification as serious eye damaging (CLP/EPA/GHS (Cat 1)). Relative to the negative control, the test item did not cause an increase of the corneal opacity or permeability. The calculated mean in vitro irritancy score was 0.50. According to OECD 437 the test item is considered as not requiring classification for eye irritation or serious eye damage.

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1985-03

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 2.4 - 2.6 kg
- Housing: individually
- Diet: ad libitum rabbit standard diet (Muemmel Z, Plange)
- Water: ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature: 23 +/- 1 °C
- Humidity (%): 45-55
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:

other: vaseline

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.1 g
- Concentration: 1 % in vaseline

Duration of treatment / exposure:

one single treatment

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

6

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: On three animals the treated eye was washed out with lukewarm water
- Time after start of exposure: 4 seconds after application

SCORING SYSTEM: according to the Draize scoring system

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

Animal 1-3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

mean

Remarks:

Animal 1-3

Time point:

24/48/72 h

Score:

0.11

Max. score:

2

Reversibility:

fully reversible within: 48 h

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

Animal 1-3

Time point:

24/48/72 h

Score:

0.11

Max. score:

3

Reversibility:

fully reversible within: 48 h

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: animal 1: treated eye was not rinsed

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: animal 2: treated eye was not rinsed

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: animal 3: treated eye was not rinsed

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

other: animal 1: treated eye was not rinsed

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

other: animal 2: treated eye was not rinsed

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

2

Reversibility:

fully reversible within: 48 h

Remarks on result:

other: animal 3: treated eye was not rinsed

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

3

Remarks on result:

other: animal 1: treated eye was not rinsed

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

3

Remarks on result:

other: animal 2: treated eye was not rinsed

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 h

Remarks on result:

other: animal 3: treated eye was not rinsed

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: animal 1: treated eye was not rinsed

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: animal 2: treated eye was not rinsed

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: animal 3: treated eye was not rinsed

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

Animal 4-6: treated eyes were rinsed after 4 sec

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

mean

Remarks:

Animal 4-6: treated eyes were rinsed after 4 sec

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

Animal 4-6: treated eyes were rinsed after 4 sec

Time point:

24/48/72 h

Score:

0

Max. score:

3

Interpretation of results:

GHS criteria not met

Conclusions:

The test substance at a concentration of 1 % was not irritating to the rabbits' eyes.

Executive summary:

An in vivo study for eye irritation was conducted similar to OECD 405. In this primary eye irritation test in 6 rabbits the test substance was applied at a concentration of 1 % in vaseline. 0.1 g were applied to one eye of each animal, the other eye served as control. The eyes were rinsed after 4 sec in 3 of the test animals. The eyes were observed for a period of 7 days and findings were scored according to the Draize system. No findings were made in the animals whose eyes were washed out after treatment. Mean 24/ 48/ 72 hour scores of the other three animals were: cornea: 0, 0, 0; iris: 0, 0.33, 0, conjunctival redness: 0, 0.33, 0, chemosis: 0, 0, 0. All effects observed in individual animals were fully reversible within 48 h after application. Based on these findings the substance at a concentration of 1 % was found to be non-irritating.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation:

An in vitro study was performed to assess the irritation potential of the test item by means of the Human Skin Model Test according to OECD 439. The test item did not reduce MTT (test for direct MTT reduction), and it did not change colour when mixed with deionised water (test for colour interference). Also its intrinsic colour was not intensive. Consequently, additional tests with freeze-killed or viable tissues were not necessary. Each three tissues of the human skin model EpiDerm were treated with the test item, the negative or the positive control for 60 minutes. 30.4 μL of the test item were applied to each tissue, and spread to match the surface of the tissue. 30 μL of either the negative control (DPBS) or the positive control (5% SLS) were applied to each tissue. After treatment with the negative control the absorbance values were well in the required range of the acceptability criterion of mean OD ≥ 0.8 and ≤ 2.8 for the 60 minutes treatment interval, thus assuring the quality of the tissues. Treatment with the positive control induced a sufficient decrease in the relative absorbance as compared to the negative control for the 60 minutes treatment interval, and thus assuring the validity of the test system. After treatment with the test item the mean relative absorbance value decreased to 90.6 % compared to the relative absorbance value of the negative control. This value is above the threshold for irritancy of ≤ 50 %. Therefore, the test item is not considered to possess an irritant potential. In conclusion, it can be stated that in this study and under the experimental conditions reported, the substance is not irritating to skin.

A study is available which was conducted with human volunteers to assess the skin irritating properties of the test substance applied at a concentration of 1 % in vaseline. Among the 50 volunteers, 39 persons were known to have eczema. The test substance preparation was applied to skin sites on the shoulder blades and occluded for 24 hours. Skin reactions were evaluated immediately after removing the patches and 24 hours thereafter. As a result, no reactions were observed. Thus, the test substance at a concentration of 1 % was shown not to be irritating to skin. Please refer to IUCLID section 7.10.5 for details.

Conclusion:

An existing skin irritation study on human volunteers showed that the test substance at a concentration of 1 % was shown not to be irritating to skin. As the substance was applied in a diluted form an in vitro test was conducted using undiluted test substance. The in vitro study according to OECD 439 determined that the undiluted test substance does not induce skin irritation. As both the human and the in vitro study concluded that the test substance does not induce skin irritation or corrosion these results are considered to be sufficient for the classification of the test substance as not skin irritating. 

 

Eye irritation

An in vitro study was performed to assess the corneal damage potential of the test substance by means of the BCOP assay using fresh bovine corneae. The study was conducted according to OECD 437. After a first opacity measurement of the fresh bovine corneae (t0), the neat test item, the positive, and the negative controls were applied to corneae fixed in an incubation chamber in horizontal position for 10 minutes at 32 ± 1 °C. The posterior chamber contained incubation medium. After the incubation phase the test item, the positive, and the negative controls were each rinsed from the corneae. Further, the corneae were incubated for another 120 minutes at 32 ± 1 °C in a vertical position, while the anterior chamber contain incubation medium as well. Afterwards, opacity was measured a second time (t130). After the opacity measurements permeability of the corneae was determined by measuring spectrophotometrically the transfer of sodium fluorescein after incubation in a horizontal position for 90 minutes at 32 ± 1 °C. With the negative control (0.9 % (w/v) NaCl solution in deionised water) neither an increase of opacity nor permeability of the corneae could be observed. The positive control (2-Ethoxyethanol) showed clear opacity and distinctive permeability of the corneae corresponding to a classification as serious eye damaging (CLP/EPA/GHS (Cat 1)). Relative to the negative control, the test item did not cause an increase of the corneal opacity or permeability. The calculated mean in vitro irritancy score was 0.50. According to OECD 437 the test item is considered as not requiring classification for eye irritation or serious eye damage.

An in vivo study for eye irritation was conducted similar to OECD 405. In this primary eye irritation test in 6 rabbits the test substance was applied at a concentration of 1 % in vaseline. 0.1 g were applied to one eye of each animal, the other eye served as control. The eyes were rinsed after 4 sec in 3 of the test animals. The eyes were observed for a period of 7 days and findings were scored according to the Draize system. No findings were made in the animals whose eyes were washed out after treatment. Mean 24/ 48/ 72 hour scores of the other three animals were: cornea: 0, 0, 0; iris: 0, 0.33, 0, conjunctival redness: 0, 0.33, 0, chemosis: 0, 0, 0. All effects observed in individual animals were fully reversible within 48 h after application. Based on these findings the substance at a concentration of 1 % was found to be non-irritating.

Conclusion:

An in vivo study comparable to OECD 405 was available that determined that the test substance was not irritating or damaging to the eyes. As only a concentration of 1 % test substance was applied an in vitro study was performed to evaluate the undiluted test substance. The in vitro OECD 437 study (BCOP) determined that the undiluted test substance does not induce eye irritation or serious damage to the eyes. Considering the results of in vivo and in vitro study, in can be concluded that the test substance does not induce eye irritation or serious damage to the eyes.

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008

Based on available data on skin irritation/corrosion, the test item is not classified according to Regulation (EC) No 1272/2008 (CLP), as amended for the ninth time in Regulation (EU) No 2016/1179.

Based on available data on eye irritation/serious eye damage, the test item is not classified according to Regulation (EC) No 1272/2008 (CLP), as amended for the ninth time in Regulation (EU) No 2016/1179.