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Description of key information

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
04 Jul - 06 Jul 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Version / remarks:
The test item was found to have the potential to cause color interference with the MTT endpoint, therefore additional color correction tissues were incorporated into the testing procedure. However, the results obtained showed that no color interference occurred.
Deviations:
yes
Remarks:
This deviation was considered to have not affected the integrity or validity of the study.
Qualifier:
according to
Guideline:
EU Method B.40 (In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER))
Version / remarks:
The test item was found to have the potential to cause color interference with the MTT endpoint, therefore additional color correction tissues were incorporated into the testing procedure. However, the results obtained showed that no color interference occurred.
Deviations:
yes
Remarks:
This deviation was considered to have not affected the integrity or validity of the study.
GLP compliance:
yes (incl. certificate)
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
Identification: Lumière Pink S.M. 8135N
Batch: 021340
Purity: >90%
Physical state/Appearance: Red powder
Expiry Date: 01 July 2022
Storage Conditions: Room temperature in the dark

Negative Control
Identification: sterile distilled water
Batch 3012436
Purity: Not supplied
Expiry Date: 01 October 2018
Storage Conditions: Room temperature

Positive Control
Identification: 8.0N Potassium Hydroxide
Batch: SLBM9898V
Purity: 7.92N
Expiry Date: 07 April 2020
Storage Conditions: Room temperature

EpiDerm™ Reconstructed Human Epidermis Model Kit
Supplier: MatTek
Date received: 04 July 2017
EpiDermTM Tissues (0.63cm2) lot number: 25828
Assay Medium lot number: 062917TMA
Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Source strain:
other: epithelial, derived from human skin, and formed into a stratified, cornified epithelium
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
Supplier: MatTek
Date received: 04 July 2017
EpiDermTM Tissues (0.63cm2) lot number: 25828
Assay Medium lot number: 062917TMA

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37°C
- Temperature of post-treatment incubation: 37°C

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: Rinsing was achieved by filling and emptying each tissue under a constant soft stream of DPBS to gently remove any residual test item.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1.0 mg/mL
- Incubation time: 60
- Optical density – 570nm
Control samples:
yes, concurrent negative control
yes, concurrent positive control
yes, concurrent MTT non-specific colour control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit):25 mg

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 50 μL


POSITIVE CONTROL
- Amount(s) applied (volume or weight): 50 μL
- Concentration (if solution): 8.0 N

25 μL of sterile water was added for wetting of the test item to increase tissue surface contact.
Duration of treatment / exposure:
3 and 60 minutes
Duration of post-treatment incubation (if applicable):
3 h at 37 °C, 5% CO2
Number of replicates:
2
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Nean Negative Control 3 min
Value:
100
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Mean Negative Control 30 min
Value:
100
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Mean Positive Control 3 min
Value:
5.6
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Mean Positive Control 60 min
Value:
4
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Mean Test Item 3 min
Value:
120.4
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Mean Test Item 60 min
Value:
100.1
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Other effects / acceptance of results:
- OTHER EFFECTS:
- Direct-MTT reduction:
- Colour interference with MTT:
The solution containing the test item was a purple color, therefore additional color correction tissues were incorporated into the testing procedure. However, the results obtained showed that no color interference occurred.


ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
- Acceptance criteria met for variability between replicate measurements:yes

The relative mean viabilities for each treatment group :

Exposure Period

Percentage Viability

Negative Control

Positive Control

Test Item

3 minute

100

5.6

120.4

60 minute

100

4

100.1

 

Mean OD570Values and Viabilities for the Negative Control Item, Positive Control Item and Test Item

Tissue

Exposure Period

Mean OD570 of individual tissues

Mean OD570 of duplicate tissues

Standard Deviation

Coefficient of Variation (%)

Relative Mean Viability (%)

Negative Control

3 Minutes

1.524

1.568

0.062

4

100

1.612

60 Minutes

1.759

1.795

0.051

2.8

1.831

Positive Control

3 Minutes

0.097

0.088

0.013

N/A

5.6

0.079

60 Minutes

0.071

0.072

0.001

N/A

4

0.073

Test Item

3 Minutes

1.796

1.888

0.13

6.9

120.4

1.98

60 Minutes

1.777

1.796

0.026

1.5

100.1

Interpretation of results:
GHS criteria not met
Conclusions:
The test item was considered to be non-corrosive to the 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
Study period:
25 Mar 1996 - 17 Apr 1996
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: LUMIERE PINK 8135C
- Storage: room temperature
- Description: dark maroon powder
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Age at study initiation: 12 – 16 weeks old
- Weight at study initiation: 2.54 – 2.73 kg
- Housing: suspended metal cages
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 - 21
- Humidity (%): 45 - 69
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/ 12
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 (=49 mg)
Duration of post- treatment incubation (in vitro):
1, 24, 48 and 72 hours, additional observatios were made on Days 7, 14 and 21
Number of animals or in vitro replicates:
3
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): N/A
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.3
Max. score:
1
Reversibility:
fully reversible within: 72h
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
0
Reversibility:
other: no opacity noted
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1
Max. score:
3
Reversibility:
not fully reversible within: 21 days
Remarks on result:
positive indication of irritation
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
0
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
0
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1
Max. score:
1
Reversibility:
fully reversible
Remarks:
14 days
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1.3
Max. score:
2
Reversibility:
fully reversible within: 7 days
Remarks on result:
probability of weak irritation
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24/48 h
Score:
1
Max. score:
1
Reversibility:
fully reversible within: 72h
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2
Max. score:
2
Reversibility:
fully reversible within: 21 days
Remarks on result:
probability of mild irritation
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1
Max. score:
1
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48 h
Score:
1
Max. score:
1
Reversibility:
fully reversible within: 72h
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2
Max. score:
2
Reversibility:
fully reversible within: 21 days

 animal #1                    animal #2           animal #3  
 24 h  48 h  72 h  mean animal #1  24 h  48 h  72 h  mean animal #2  24 h  48 h 72 h   mean animal #3

 mean

Conjunctival Redness

2

1

1

1.3

1

1

0

0.7

2

 2

2

2

1.3

Conjunctival Chemosis

1

1

1

1

1

1

0

0.7

2

2

2

2

0.9

Iritis 0 0 0 0 0 0 0 0 1 1 1 1 0.3

Corneal opacity

1

0

0

0.3

0

0

0

0

1

1

1

1

0.6
Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Conclusions:
The test item was found to irritating to rabbit eye.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification

Test substance is not classified for irritation to the skin but is classified as a Cat 1 eye irritant.