3,5,5-trimethylhexyl acetate

Endpoint summary

Description of key information

The registration item is considered as non-irritating to skin and non-irritating to eyes.

 

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2020-11-12 to 2020-12-11

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method B.40 (In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER))

Version / remarks:

30-05-2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: MatTek test protocol: In vitro EpiDerm Skin Corrosion Test (EPI-200-SCT)

Version / remarks:

2014-11-07

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)

Version / remarks:

adopted 2019-06-18

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test system:

human skin model

Source species:

other: reconstructed human epidermis

Cell type:

non-transformed keratinocytes

Cell source:

other: MatTek Corporation (Bratislava, Slovakia)

Source strain:

other: not applicable

Justification for test system used:

The epidermis model (EpiDerm™) is derived from human keratinocytes and consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multilayered, highly differentiated model of the human epidermis.

Validation studies have shown that tests employing human skin models are able to discriminate between known skin corrosives (Optional subcategory 1A, optional subcategory 1B and 1C) and non-corrosives.

Laboratory technical proficiency with the test system according to OECD Test Guideline 431 was demonstrated at ICCR - Roßdorf GmbH in December 2019.

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm™
- Tissue batch number: 34108
- Date of initiation of testing: 2020-11-11

TEMPERATURE USED FOR TEST SYSTEM
- 3 min at room temperature and 60 min at 37°C, 5% CO2, 95% room humidity

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: 20 times with a constant soft stream of 1x PBS


MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL
- Incubation time: 3 h (± 5 min)
- Wavelength: 570 nm
- Spectrophotometer: Versamax® Molecular Devices; Software: SoftMax Pro Enterprise

NUMBER OF REPLICATE TISSUES: 2

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

PREDICTION MODEL / DECISION CRITERIA
-The test substance is predicted to be corrosive to skin if the viability measured after 3 minutes is less than 50%, or the viability is less or equal than 50% after 3 minutes of exposure AND less than 15% after 60 minutes of exposure.
- The test substance is considered to be corrosive to skin (Cat. 1A) if the viability after 3 minutes exposure is less than 25%.
- The test substance is considered to be corrosive to skin (Cat 1B and 1C) if the viability after 3 minutes exposure is greater than or equal to 25%

- The test substance is considered to be non-corrosive to skin if the viability is greater than or equal to 50% after 3 minutes of exposure AND greater than or equal to 15% after 60 minutes of exposure.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

50 µL (79.4 µL/cm2 according to guideline) for test material and controls

Duration of treatment / exposure:

3 ± 0.5 minutes and 60 ± 5 minutes

Number of replicates:

2

Irritation / corrosion parameter:

% tissue viability

Remarks:

mean of 2 replicates after 60 minutes exposure

Run / experiment:

1

Value:

103.51

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: no indication of corrosion

Irritation / corrosion parameter:

% tissue viability

Remarks:

mean of 2 replicates after 3 minutes exposure

Run / experiment:

1

Value:

101.6

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: no indication to corrosion

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: No
- Direct-MTT reduction: No
- Colour interference with MTT: No


DEMONSTRATION OF TECHNICAL PROFICIENCY: Yes

ACCEPTANCE OF RESULTS:
- Acceptance criterion 1. met for negative control: Yes (mean OD: 1.906 [3 min], 1.913 [60 min])
- Acceptance criterion 2. met for positive control: Yes (mean Rel. Viability [%] after 60 min of exposure: 4.01)
- Acceptance criterion 3. met for variability between replicate measurements: Yes (Coefficient of Variation (CV): 4,0% [3 min], 3.1 [60 min])

Results after treatment with the test item and the controls after 3 minutes exposure 

Treatment Group	Tissue No.	Exposure Interval [min]	OD 570 mm			Mean OD of 3 wells blank corrected	Mean OD of 2 tissues blank corrected	Standard deviation (well 1. 2 and 3)	Mean Rel. Viability [%]*	CV [%]
			Well 1	Well 2	Well 3
Blank		3	0.039	0.040	0.044
Negative Control	1		0.049	1.967	1.950	1.984	1.906	0.053	100.00	3.1
	2		2.049	1.967	1.950	1.948		0.024
Positive Control	1		0.490	0.458	0.463	0.430	0.369	0.018	19.35	23.3
	2		0.356	0.344	0.346	0.308		0.006
Test Item	1		2.079	2.004	2.013	1.991	1.936	0.041	101.60	4.0
	2		1.980	1.898	1.887	1.881		0.051

* mean Viability (%) = 100 x mean OD TI/PC/NC 
mean OD NC 

Results after treatment with the test item and the controls after 60 minutes exposure 

Treatment Group	Tissue No.	Exposure Interval [min]	OD 570 mm			Mean OD of 3 wells blank corrected	Mean OD of 2 tissues blank corrected	Standard deviation (well 1. 2 and 3)	Mean Rel. Viability [%]*	CV [%]
			Well 1	Well 2	Well 3
Blank		60	0.039	0.040	0.044
Negative Control	1		2.011	1.927	1.926	1.914	1.913	0.049	100.00	0.1
	2		2.007	1.933	1.919	1.912		0.047
Positive Control	1		0.148	0.133	0.121	0.093	0.077	0.013	40.1	30.5
	2		0.103	0.101	0.099	0.060		0.002
Test Item	1		2.012	1.963	1.957	1.937	1.980	0.030	103.51	3.1
	2		2.111	2.048	2.035	2.024		0.040

* mean Viability (%) = 100 x mean OD TI/PC/NC 
mean OD NC 

 

Interpretation of results:

GHS criteria not met

Conclusions:

In conclusion, it can be stated that in this study and under the reported experimental condition the test item is non-corrosive to skin according to EU CLP and UN GHS.

Executive summary:

This in vitro study was performed to assess the corrosive potential of the test item by means of the Human Skin Model Test with EpiDerm™ tissues models.
The test item did not reduce MTT (pre-test for direct MTT reduction), and it did not change colour when mixed with deionised water and isopropanol (pre-test for colour interference). Consequently, additional tests with freeze-killed tissues, viable tissues and non-specific killed tissues to determine correction factors for calculating the true viability in the main experiment were not necessary.
Independent duplicate tissues of EpiDerm™ were exposed to either the test item, the negative control (Dulbecco's Phosphate-Buffered Saline (DPBS)) or the positive control (8.0 N KOH) for 3 minutes and 1 hour, respectively.
After exposure to the negative control the absorbance values met the required acceptability criterion of a mean OD570 ≥ 0.8 and ≤ 2.8 for both treatment intervals thereby confirming the acceptable quality of the tissues.
Exposure to the positive control induced a decrease in the relative absorbance as compared to the negative control, both for the 3 minutes exposure period and for the 1 hour exposure period. The 1 hour exposure caused a decrease of the cell viability < 15% of the negative control. The CV in the range 20 – 100% viability between the tissue replicates is ≤ 30%, thus the validity of the test system and the specific batch of the tissue models is confirmed.
After exposure of the tissues to the test item the relative absorbance value was 101.60% after 3 minutes exposure. After 1 hour exposure the relative absorbance value was 103.51%. Both values did not reach the threshold for corrosivity which is defined to be < 50% after the 3 minutes exposure or ≥ 50% after 3 minutes exposure and < 15% after the 1 hour exposure.
Therefore, the test item is considered to be non-corrosive.

Reference 2

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2021-01-12 to 2021-02-05

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: MatTek Corporation Protocol: In vitro EpiDermTM Skin Irritation Test (EPI-200-SIT) for use with MatTek Corporation’s Reconstructed Human Epidermal Model EpiDerm (EPI-200).

Version / remarks:

2019-02-10

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2012-07-06

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted 2019-06-18, corrected 2020-06-26

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: MatTek Corporation (82105 Bratislava, Slovakia)

Source strain:

other: Not applicable

Justification for test system used:

The EpiDerm™ tissue consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multilayered, highly differentiated model of
the human epidermis. The EpiDerm™ Skin Irritation Test (SIT) was validated in an international prevalidation study performed by ECVAM and have turned out as a sufficiently promising predictor for skin irritancy potential. Laboratory technical proficiency with the test system according to OECD Test Guideline 439 was demonstrated at ICCR-Roßdorf GmbH in March 2014.

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm™
- Tissue batch number(s):
- Date of initiation of testing: 2021-02-05

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 ± 1.5°C and 5 ± 0.5% CO2 for 35 minutes, remaining 25 minutes at room temperature
- Temperature of post-treatment incubation: 37 ± 1.5 °C and 5 ± 0.5% CO2

REMOVAL OF TEST MATERIAL AND CONTROLS
Tissues were gently rinsed with PBS several times in order to remove any residual test material.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL in DMEM
- Incubation time: 3 hour ± 5 minutes
- Spectrophotometer: Versamax® Molecular Devices
- Wavelength: 570 nm
- Software: SoftMax Pro Enterprise (version 4.7.1)

NUMBER OF REPLICATE TISSUES: 3

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

CALCULATION
True viability = Viability of treated tissue – Interference from test chemical = ODtreated viable tissue - ODkilled tissue, where ODkilled tissue = (mean ODtreated killed tissue - mean ODuntreated killed tissue)

PREDICTION MODEL / DECISION CRITERIA
- The test substance is considered to be a skin irritant if the viability after 3 hours exposure is less than or equal 50%.
- The test substance is considered to be non-irritant to skin if the viability after 3 hours exposure is greater than 50%.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

30 µL (47 µL/cm2 according to guideline) for controls and test item

TEST MATERIAL
- Amount applied: 30 µL (undiluted)

NEGATIVE CONTROL
- Amount applied: 30 µL (undiluted)

POSITIVE CONTROL
- Amount applied: 30 µL
- Concentration: 5 %

Duration of treatment / exposure:

60 minutes (35 minutes at 37°C and 25 minutes at room temperature)

Duration of post-treatment incubation (if applicable):

42 hours (incubation medium was changed after 24 hours)

Number of replicates:

3

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Mean of three replicates

Value:

90.19

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

- OTHER EFFECTS:
- Visible damage on test system: Not indicated
- Direct-MTT reduction: The test item did not prove to be a MTT reducer in the MTT interference pre-experiment.
- 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 lead to a change in colour.

DEMONSTRATION OF TECHNICAL PROFICIENCY: Yes, see "Attached background material"

ACCEPTANCE OF RESULTS:
- Acceptance criteria (1.) met for negative control: Yes (mean OD: 1.584)
- Acceptance criteria (2.) met for positive control: Yes (mean rel. viability 4.87)
- Acceptance criteria (3.) met for the standard deviation: Yes (SD[NC]: 4.5, SD[PC]: 0.4, SD[TI]:1.1)
- Acceptance criteria (4.) met for historical acceptance range

Results after treatment with the test item and the controls 

Treatment Group	Tissue No.	OD 570 nm			Mean OD of 3 Wells	Mean OD of 3 Wells blank corrected	Mean OD of 3 tissues	Rel. Viability [%] Tissue 1.2 +3	Standard Deviation	Mean Rel. Viability [%]
		Well 1	Well 2	Well 3
Blank		0.040	0.040	0.040	0.040
Negative Control	1	1.732	1.701	1.675	1.703	1.662	1.584	104.937	4.5	100.0
	2	1590	1.559	1.536	1.562	1.521		96.035
	3	1.634	1.604	1.589	1.609	1.569		99.029
Positive Control	1	0.127	0.122	0.122	0.124	0.084	0.077	5.273	0.4	4.87
	2	0.115	0.117	0.118	0.117	0.077		4.831
	3	0.111	0.111	0.112	0.111	0.071		4.498
Test Item	1	1.504	1.437	1.430	1.457	1.417	1.429	89.430	1.1	90.19
	2	1.467	1.454	1.460	1.460	1.420		89.638
	3	1.499	1.489	1.481	1.490	1.450		91.496

 

Interpretation of results:

GHS criteria not met

Conclusions:

In conclusion, it can be stated that in this study and under the experimental conditions reported, the test item is non-irritant to skin according to UN GHS and EU CLP regulation.

Executive summary:

This in vitro study was performed to assess the irritation potential of the test item by means of the Human Skin Model Test. The test item did not prove to be a MTT reducer in the MTT interference pre-experiment. Also, its intrinsic color was not intensive, and it did not change color when mixed with deionised water or isopropanol. Therefore, additional tests with freeze-killed tissues or viable tissues (without MTT addition) did not have to be performed. Three tissues each of the human skin model EpiDerm™ were treated with the test item, the negative control (DPBS) or the positive control (5% SDS) for 60 minutes. After treatment with the negative control the absorbance values were well within 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 viability as compared to the negative control for the 60 minutes treatment interval, thus assuring the validity of the test system. After treatment with the test item the mean relative viability value was 90.19% 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 test item is non-irritant to skin.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

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:

yes

Remarks:

The negative response observed in the animal treated with undiluted substance was not confirmed by testing 2 additional animals.

Principles of method if other than guideline:

The method used to assess eye irritancy is a modification of that prescribed for purposes of the United States Federal Hazardous Substances Labelling Act. In addition corneal damage is also assessed objectively by measurement of corneal swelling.
3 rabbits were treated with 100 µL of a 10 % dilution, 1 rabbit was treated with 100 µL of a 50 % dilution and 1 rabbit was treated with 100 µL undiluted substance. No additional rabbits were tested to confirm the negative results observed for the undiluted substance.

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: commercial supplier
- Age at study initiation: >= 7 weeks
- Weight at study initiation: >= 1 kg
- Housing: singly in cages from which all hay, straw or similar potentially irritant material is excluded
- Water: ad libitum
- Acclimation period: yes, duration not reported

Vehicle:

other: Tween 80

Controls:

other: Left eye served as untreated control.

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 100 µL
- Concentration: 10 % (3 rabbits), 50 % (1 rabbit), 100 % (1 rabbit)

Observation period (in vivo):

Conjunctival damage: 15 minutes after treatment
Corneal, conjuntival and iridial reaction: 1, 2, 3, 4, 7, 9, 11, 16, 18 and 21 days after treatment. The assessment is stopped earlier if the eye returned to normal.

Number of animals or in vitro replicates:

Unchanged substance: 1 rabbit
50 % in Tween 80: 1 rabbit
10 % in Tween 80: 3 rabbits

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: No

SCORING SYSTEM:
CORNEA
0: Normal
0.5: Any change form normal, including slight dulling of corneal lustre
1: Scattered or diffuse areas of opacity, or loss of corneal epithelium. Iris clearly visible
2: Easily discernible, greyish, translucent areas, details of iris slightly obscured
3: Grey-white areas, no details of iris visible, size of pupil barely discernible
4: Complete corneal opacity, iris not discernible

IRIS
0: Normal
+: Any divergence from normal, congestion of iridial vessels, deepening of folds
++: Severe congestion of iridial vessels or deepening of folds. Loss of iridial reflex to light

AREA OF CORNEA OR IRIS AFFECTED
1: Less than 1/4 of area affected
2: Between 1/4 and 1/2 of area affected
3: Between 1/2 and 3/4 of area affected
4: Between 3/4 and whole area affected

CONJUNCTIVAE
Redness (palpebral and bulbar conjunctivae and nictitating membrane)
0: Vessels normal
1: Some vessels definitely injected
2: Diffuse, crimson red, individual vessels not easily discernible
3: Diffuse, beefy red

Chemosis
0: No swelling
1: Any swelling above normal
2: Obvious swelling with partial eversion of eyelids
3: Swelling with lids about half closed
4: Swelling with lids more than half closed

Discharge
0: No discharge
1: Any discharge different from normal
2: Discharge with moistening of lids and hairs just adjacent to the lids
3: Discharge with moistening of lids and considerable area round eye

TOOL USED TO ASSESS SCORE: hand torch

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Remarks on result:

other: 100 µL undiluted test substance

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

fully reversible

Remarks on result:

other: 100 µL undiluted test substance

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible

Remarks on result:

other: 100 µL undiluted test substance

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.67

Max. score:

4

Reversibility:

fully reversible

Remarks:

5 days

Remarks on result:

other: 100 µL undiluted test substance

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.66

Max. score:

4

Reversibility:

fully reversible within: 5 days

Remarks on result:

other: 100 µL 50 % in Tween 80

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

fully reversible

Remarks on result:

other: 100 µL 50 % in Tween 80

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 5 days

Remarks on result:

other: 100 µL 50 % in Tween 80

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.66

Max. score:

4

Reversibility:

fully reversible within: 4 days

Remarks on result:

other: 100 µL 50 % in Tween 80

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 5 days

Remarks on result:

other: 100 µL 10 % Tween 80

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

fully reversible

Remarks on result:

other: 100 µL 10 % Tween 80

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

fully reversible within: 5 days

Remarks on result:

other: 100 µL 10 % Tween 80

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 4 days

Remarks on result:

other: 100 µL 10 % Tween 80

Interpretation of results:

GHS criteria not met

Conclusions:

According to the scores obtained for the undiluted substance, the substance has a low potential to be irritating to the eyes. However, it has to be considered that only one animal was dosed with undiluted substances instead of three as instructed in OECD TG 405.

Executive summary:

The potential of 3,5,5 -trimethylhexyl acetate to induce eye irritation was tested in a screening test performed according to the modified Federal Hazardous Substances Labelling Act method. Three rabbits were treated with 100 µL of a 10% dilution of the substance in Tween 80, one rabbit was treated with 100 µL of a 50% dilution in Tween 80 and one rabbit with 100 µL undiluted substance. In the eyes treated with 10% dilution, only slight superficial corneal imperfections were caused that persisted for three to five days. Conjunctivitis was also slight and persisted for two to four days. In the eye treated with 50% dilution, a loss of 1/4 or corneal epithelium was seen, with slight corneal swelling. Corneal repair was complete after 5 days and the moderate conjuntivitis with oedema also healed after 5 days. In the eye treated with undiluted substance, the cornea remained unaffected. Only slight conjunctivitis was seen which persisted for four days.

The numerical grading of the ocular lesions was equivalent to the grading suggested in OECD TG 405. The mean of the cornea, iris and chemosis scores at 24, 48 and 72 h was below 1 in all animals and the mean of the conjunctivae scores at 24, 48 and 72 h was below or equal to 1.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation/ corrosion

Skin irritation/ corrosion potential of the registration item were evaluated in a weight of evidence approach using reconstructed human epidermis (RHE) based in vitro methods according to OECD guidelines no. 431 and 439.

Skin corrosion potential was assessed according to OECD guideline no. 431. After exposure of the tissues to the test item the relative absorbance value was 101.60% after 3 minutes exposure. After 1 hour exposure the relative absorbance value was 103.51%. Both values did not reach the threshold for corrosivity which is defined to be < 50% after the 3 minutes exposure or ≥ 50% after 3 minutes exposure and < 15% after the 1 hour exposure. Therefore, the test item is considered to be non-corrosive.

Skin irritation potential was assessed according to OECD guideline no. 439. After treatment with the test item the mean relative cell viability value was 90.19% 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.

 

The stated results justificate the consideration of the test item as non-irritating.

 

 

Eye irritation / corrosion

Two in vivo studies on eye irritation are available. In one of the studies, eight rabbits were treated with the test article (0.1 mL) diluted to 50% in olive oil. The eyes of 4 rabbits were rinsed immediately after dosing; the other 4 rabbits were left without rinsing. No mucosal reactions in the area of the eyes or the mucosa of the eye were noted. The spontaneous defence actions were insignificant.

In the second study, three rabbits were treated with 100 µL of a 10% dilution in Tween 80, one rabbit was treated with 100 µL of a 50% dilution in Tween 80 and one rabbit was treated with 100 µL undiluted substance. The grading of the ocular lesions was equivalent to the grading suggested in OECD TG 405. The mean of the cornea, iris and chemosis scores at 24, 48 and 72 h was below 1 in all animals and the mean of the conjunctivae scores at 24, 48 and 72 h was below or equal to 1 in all animals.

Thus, the available data set has certain limitations with regard to study quality and description. As all studies revealed low irritation potential and in order to respect the animal testing ban for cosmetics, it is still proposed to waive further in vivo testing and to conduct the hazard assessment on the basis of the available data.

Justification for classification or non-classification

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

 

Skin irritation / corrosion

The available experimental test data are reliable and suitable for non-classification purposes under Regulation (EC) No 1272/2008. Based on available data on skin irritation/corrosion, the test item is classified and labelled as non-irritating to skin according to EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No 1272/2008, as amended for fifteenth time in Regulation (EU) No 2020/217.

 

Eye irritation / corrosion

The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. Based on available weight of evidence data on eye irritation/corrosion, the test item is not classified according to Regulation (EC) No 1272/2008 (CLP), as amended for the fifteenth time in Regulation (EU) No 2020/217.