Registration Dossier

Administrative data

Description of key information

The substance was tested for skin irritation in the EpiDermTM model according to OECD 439. 30 uL of the substance, positive control (5% SDS) and negative control (PBS) was applied to the tissue for 60 minutes (25 minutes at room temperature and 35 minutes at 37°C, 5% CO2) and thereafter incubated for 42 hour post-dose at 37°C, 5% CO2. The substance did not interfere by colour or by direct MTT reduction. Tissue viability was 106.9% of negative controls, while validity criteria were evaluated to be met. The substance is concluded to be non-irritant.

The substance was tested for eye irritation in the EpiOcular TM  Eye Irritation model according to OECD 492. 50 uL of the substance, positive control (methyl acetate) and negative control (H2O) was applied to the tissue for 30 minutes, followed by a 12 minutes post-treatment immersion at 37°C, 5% CO2 and thereafter incubated for 2 hour post-dose at 37°C, 5% CO2. 

The substance did not interfere by colour or by direct MTT reduction. Tissue viability was 100.1% of negative controls, while validity criteria were met. The substance is concluded to be non-irritant.

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:
07 December 2017 to 15 December 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
GLP compliance:
yes (incl. certificate)
Test system:
human skin model
Source species:
human
Cell type:
other: reconstructed human skin
Cell source:
other: EpiDerm TM (EPI-200-MatTek Corporation)
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm TM (EPI-200-MatTek Corporation)
- Tissue batch number(s): (Lot 25864)
- Delivery date: 12 December 2017
- Date of initiation of testing: no data

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 60 minutes (25 minutes at room temperature and 35 minutes at 37°C, 5% CO2)
- Temperature of post-treatment incubation: 42 hour post-dose incubation at 37°C, 5% CO2

REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps: not indicated (according to MatTEek SOP)

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE: no data (according to MatTEek SOP)
- MTT concentration: not specified
- Incubation time: no data
- Spectrophotometer: not identified measured at 570 nm
- Wavelength: 570 nm
- Filter: none
- Linear OD range of spectrophotometer: no data

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: yes (1.601 ± 0.007 (mean and SD of MTT value of 3 tissues exposed to H2O) )
- Barrier function: yes (ET 50% of tissues exposed to 100 µL Triton X-100 1% (n=3): 5.84 h)
- Morphology: yes ( Histological examination demonstrates human epidermis-like structure: including multiple layers (at least 4) of viable epithelial cells (basal layer, stratum spinosum, stratum granulosum) which are present under multilayered stratum corneum ; Tissue thickness: 128 µm )
- Contamination: No contamination reported
- Reproducibility: yes (The mean OD of the 6 wells containing extraction solvent alone (blanks) should be ≤ 0.1., but was 0.149 evaluated not to have affected the measurements, based upon historical system data for blanks; < 0.244)
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 30 µl

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 30 µl PBS

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 30 µl SDS
- Concentration (if solution): 5% in water
Duration of treatment / exposure:
60 minutes (25 minutes at room temperature and 35 minutes at 37°C, 5% CO2)
Duration of post-treatment incubation (if applicable):
42 hour post-dose incubation at 37°C, 5% CO2
Number of replicates:
3/treatment
Irritation / corrosion parameter:
% tissue viability
Value:
106.9
Negative controls validity:
valid
Positive controls validity:
valid
Remarks:
mean viability 4.5%
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
No colour interference or interference with MTT reduction as tested in a pre-test

Name

Code

Mean of OD

SD of OD

Mean of viability (%)

SD of viability (%)

CV %

Classification

DPBS

NC

1.768

0.210

100.000

11.904

11.904

Non-Irritant

SDS 5%

PC

0.080

0.016

4.535

0.909

20.046

Irritant

Substance

TA2

1.889

0.142

106.873

8.008

7.493

Non-Irritant

 

 

Interpretation of results:
GHS criteria not met
Conclusions:
The substance is non-irritant in the EpiDermTM model
Executive summary:

The substance was tested for skin irritation in the EpiDermTM model according to OECD 439. 30 uL of the substance, positive control (5% SDS) and negative control (PBS) was applied to the tissue for 60 minutes (25 minutes at room temperature and 35 minutes at 37°C, 5% CO2) and thereafter incubated for 42 hour post-dose at 37°C, 5% CO2. The substance did not interfere by colour or by direct MTT reduction. Tissue viability was 106.9% of negative controls, while validity criteria were evaluated to be met. The substance is concluded to be non-irritant.

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:
05 December 2017 to 14 December 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)
GLP compliance:
yes (incl. certificate)
Species:
human
Details on test animals or tissues and environmental conditions:
- Tissue :Reconstructed human Cornea-like Epithelium (RhCE)
- Model used: ‘EpiOcularTM irritation test ( MatTek Corporation EpiOcularTM EIT (OCL-200) Reconstituted Human Ocular Epithelium )
- Tissue batch number(s): (Lot # 27017)
- Delivery date: 12 December 2017
- Date of initiation of testing: no data

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 30 minutes ± 2 minutes, followed by a 12 ± 2 minutes’ post-treatment immersion at 37°C, 5% CO2
- Temperature of post-treatment incubation: 2 hours ± 15 minutes’ post-treatment incubation at 37°C, 5% CO2

REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps: not indicated (according to MatTEek SOP)

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE: no data (according to MatTek SOP)
- MTT concentration: not specified
- Incubation time: no data
- Spectrophotometer: not identified measured at 570 nm
Vehicle:
unchanged (no vehicle)
Controls:
yes
yes, concurrent positive control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 50 µl
Duration of treatment / exposure:
30 minutes ± 2 minutes, followed by a 12 ± 2 minutes’ post-treatment immersion at 37°C, 5% CO2
Number of animals or in vitro replicates:
3 replicates
Details on study design:
tisues were wetted with 20 uL PBS before application of the substance or controls

negative control: sterile H2O
positive control: methyl acetate (neat)

number of replicates: 3
Irritation parameter:
other: viability (%)
Value:
100.1
Negative controls validity:
valid
Remarks:
OD570 1.992
Positive controls validity:
valid
Remarks:
viability 40.03%
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
Acceptance criteria passed

Acceptance criterion 1 The mean OD570 of the negative control(treated with PBS) tissues is > 0.8 and < 2.8
 
Acceptance criterion 2 The mean of the positive control relative percentage viability must be below 50% of the mean of the negative controls.
 
Acceptance criterion 3 The standard deviationof OD values between three replicates in each experimental condition must be ≤ 18%
 

Name

Code

mean of OD

SD of OD

Mean of viability (%)

SD of viability (%)

CV %

Classification

Sterile water

NC

1.992

0.094

100.000

4.74

4.738

Non-Irritant

Methyl Acetate

PC

0.797

0.139

40.030

6.966

17.401

Irritant

Substance

TA1

1.994

0.049

100.100

2.467

2.465

No Category

Interpretation of results:
GHS criteria not met
Conclusions:
The substance is not irritant when tested in the EpiOcular TM Eye Irritation Test
Executive summary:

The substance was tested for eye irritation in the EpiOcular TM  Eye Irritation model according to OECD 492. 50 uL of the substance, positive control (methyl acetate) and negative control (H2O) was applied to the tissue for 30 minutes, followed by a 12 minutes post-treatment immersion at 37°C, 5% CO2 and thereafter incubated for 2 hour post-dose at 37°C, 5% CO2. 

The substance did not interfere by colour or by direct MTT reduction. Tissue viability was 100.1% of negative controls, while validity criteria were met. The substance is concluded to be non-irritant.

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

Based on the information available the substance does not need to be classified for skin- and eye irritation according to EC Regulation 1272/2008 (CLP).