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Administrative data

Description of key information

The test item was found to be not irritant in an in vitro skin irritation test according to OECD 439 and not irritant in an

in vitro eye irritation test according to OECD 437.

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
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)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Deviations:
no
Principles of method if other than guideline:
The test is performed according to OECD criteria.
GLP compliance:
yes (incl. QA statement)
Remarks:
Hessisches Minesterium für Umwelt, Klimatschutz, Landwirtschaft und Verbraucherschutz
Species:
human
Strain:
other: human reconstructed epidermis model
Details on test animals or test system and environmental conditions:
Epi-200 SIT kits and MTT-100 assays diluent from MatTek Corporation (82105 Bratislava, Slovakia).
EpiDerm tissue consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multilayered, highly differentiated model of human epidermis
Type of coverage:
other: in vitro test
Preparation of test site:
other: in vitro test
Vehicle:
unchanged (no vehicle)
Controls:
other: Negative control: 30µL DPBS (MatTek) ; Positive control: 30µL of a 5% SLS (Sodium Lauryl Sulphate) solution in deiosined water (MatTek)
Duration of treatment / exposure:
60 minutes
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Mean
Value:
96.5
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation

Results:

Dose Treatment 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 60 min 1.705 1.552 1.747 1.668 102.2 6.1 100.0
93.1
104.7
Positive Control 60 min 0.075 0.083 0.085 0.081 4.5 6.4 4.9
5.0
5.1
Test item 60 min 1.537 1.663 1.630 1.610 92.2 4.0 96.5
99.7
97.7

* Mean of three replicate wells after blank correction

** Relative absorbance per tissues (rounded value): (100*absorbance of tissue)/ (mean absorvbance negative control)

*** Relative absorbance per treatment group (rounded values): (100* 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 96.5% (threshold for irritacy ≤ 50%), consequently the test item was not irritant to skin.

Interpretation of results:
not irritating
Conclusions:
In conclusion, it can be stated that in this study and under the experimental conditions reported the test substance is not irritant to skin to UN GHS and EU CLP regulation.
Executive summary:

The study has been performed according to OECD 439 guideline. This in vitro study was performed to assess the irritation properties of the test substance. by means of the Human Skin Model test.

The test item did not reduce MTT and it did not change colour when mixed with deionised water. 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 control and the positive control for 60 minutes. 30 µL of the test substance were applied to each tissues, and spread to match the surface of the tissues. 30 µL of either the negative (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 60 minutes treatment interval, and thus assuring the validity of the test system.

After the treatment with the test substance, the mean relative absorbance value decreased irrelevantly to 96.5% 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 substance is not irritant to skin to UN GHS and EU CLP regulation

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
20-11-2015
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)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)
Deviations:
no
GLP compliance:
yes
Remarks:
Hessisches Ministerium für Umwelt, Klimaschutz, Landwirtschaft und Verbraucherschutz
Species:
cattle
Details on test animals or tissues and environmental conditions:
- Test system: Freshly isolated bovine cornea (at least 9 month old donor cattle)
- Source: Schlachthof Aschaffenburg 63739 Aschaffenburg, Germany
Vehicle:
unchanged (no vehicle)
Remarks:
undiluted
Controls:
yes
Amount / concentration applied:
Anterior compartment received the test item or negative or positive control at volume of 0.75 mL on the surface of the corneae.
TEST MATERIAL
- Amount(s) applied (volume): 0.75 mL
- Concentration (if solution): undiluted test item

Duration of treatment / exposure:
First incubation time: lasted 10 min at 32°+/-1°C in water bath in a horizontal position
Second incubation after rinsing: 2 hours at 32+/-1°C in a vertical position
Observation period (in vivo):
Measurement of opacity via opacitometer
Measurement of permeability via spectrophotometer
Number of animals or in vitro replicates:
3 corneae/group
Details on study design:
Negative control: saline solution (0.9% NaCl in deionized water)
Positive control: 2-Ethoxyethanol (purity 99%)
Irritation parameter:
in vitro irritation score
Run / experiment:
mean
Value:
2.22
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
Please see details on results under section "Any other information on results"

Results after 10 minutes 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 0 0.00 0.052 0.064 0.78 0.96 Not categorized
0 0.085 1.28
0 0.055 0.83
Positive Control 64.00* 1744* 90.16 84.29 Category 1
60.00* 1355* 80.33
64.00* 1225* 82.38
Test Item 2.00* 0.141* 4.12 2.22 Not categorized
0.00* 0.102* 1.53
1.00* 0.000* 1.00

* Corrected values

Interpretation of results:
not irritating
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
According to the current study and under the experimental conditions reported, the test substance is not cagetorized.
Executive summary:

This in vitro study was performed to determine the corneal damage potential of the test substance according to the OECD guideline 437.

After the first opacity measurement of the fresh bovine cornea (t0), the neat test item, the positive and the negative controls were applied to the corneae and incubated for 10 minutes at 32° +/- 1°C. 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°C +/- 1°C in incubation medium, and opacity was measured a second time (t130).

After the opacity measurements the permeability of the corneea was determined by measuring spectrophotometrically the transfer of sodium fluorescein after incubation in a horizontal position for 90 minutes at 32 +/- 1°C.

The test substance was tested undiluted. Relative to the negative control, the test substance did not cause an increase of the corneal opacity or permeability. The calculated mean IVIS was 2.22 (threshold for serious eye damage: IVIS ≥ 55).

According to OECD 437 the test item is considered not irritant (and not categorized accoding to UN GHS).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification

Skin Irritation / Corrosion

In a skin irritation test according to OECD 439 the test item had a mean relative absorbance value of 96.5% which is above the threshold for irritancy of ≤ 50% and therefore, the test item is considered to possess no irritant potential. As a consequence, the substance should not be classified as irritant or corrosive to the skin.

Eye Irritation / Corrosion

In a BCOP assay using fresh bovine corneae according to OECD 437 and GLP, the mean IVIS of the test item was 2.22. According to the criteria stipulated in this test, no classification is required for eye irritation or corrosion for IVIS values ≤ 3.0.