Registration Dossier

Administrative data

Description of key information

Skin irritation

As the substance is reported as being a skin irritant/corrosive to vertebrates in literature and known to be severly irritant to the eyes and listed in Annex VI REACH as "Causes serious eye damage, Cat. 1" it was decided to assess the skin irritation potential in an in-vitro study only. The purpose of this study was to determine the skin irritation potential of the test material using the SkinEthic Reconstituted Human Epidermal model (RHE, SkinEthic Laboratories, Nice, France) following exposure periods of 4 and 24 hours. The relative mean viability of the test material treated tissues was 108.1% after 4 hours exposure and 96.7% after 24 hours exposure. In conclusion. The test material was considered to be a non-irritant in this in-vitro test.

Eye irritation

The substance is reported in literature as severe eye irritant and listed in Annex VI REACH as corrosive leading to "Causes serious eye damage, Cat. 1". Therefore, it was decided to follow this classification and to conduct no further eye irritation testing.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
June-July 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Version / remarks:
Study was performed before publication of the corresponding OECD- and EU-testing methods
Deviations:
no
Principles of method if other than guideline:
The purpose of this study was to determine the skin irritation potential of the test material using the SkinEthic Reconstituted Human Epidermal model (RHE, SkinEthic Laboratories, Nice, France) following exposure periods of 4 and 24 hours. The SkinEthic RHE model incorporates several features which make it advantageous in the study of skin irritancy potential. The model consists of an airlifted, living, multi-layered epidermal tissue construct, produced in polycarbonate inserts in serum-free and chemically defined medium, featuring normal ultra-structure and functionally equivalent to human epidermis in vivo. Test materials are applied directly to the culture surface, at air interface, so that undiluted and/or end use dilutions can be tested directly. The test is based on the hypothesis that irritant chemicals are able to penetrate the stratum corneum of the Skin Ethic RHE model and are sufficiently cytotoxic to cause cell death in the underlying cell layers.
GLP compliance:
yes (incl. certificate)
Specific details on test material used for the study:
- Sponsor's identification: LZ 6119
- Description: white powder
- Batch number: CSA0901038
- Purity: >= 99.5% (w/w)
- Date received: 01 May 2009
- Storage conditions: at room temperature in the dark
Test system:
human skin model
Remarks:
SkinEthicTM RHE is an in vitro reconstructed human epidermis from normal human keratinocytes cultured on an inert polycarbonate filter at the air-liquid interface, in a chemically defined medium.
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
other: not specified (Skin Ethic Laboratories, Nice, France)
Details on animal used as source of test system:
no animals tested in this study
Justification for test system used:
as the test item is know to be a severe skin irritant (listed in Annex I), it was decided to run an in-vitro test instead of an in-vivo study
Vehicle:
unchanged (no vehicle)
Details on test system:
- Supplier: Skin Ethic Laboratories, Nice, France
- Date received: 03 June 2009
- SkinEthic Tissues (0.5cm2): Lot Number 09 022A0506
- Maintenance Medium: Lot Number 0905 011J 398

Receipt of Tissues:
On arrival, the SkinEthic RHE tissues (Day 19), were stored at room temperature prior to transferring into 6-well plates designated 'arrival plates' containing 1 ml of maintenance
medium. It was important to ensure that there were no air bubbles present under the tissue inserts. The tissues were incubated overnight at 37"C, 5% COz in air.

Preparation of Tissues:
Using sterile techniques, 800 ul of maintenance medium at room temperature, was dispensed into the appropriate number of wells of 6-well plates designated 'treatment plates'. Each well was labelled with details of the treatment and the appropriate exposure time. Separate treatment plates were used for each substance and negative control to avoid the possibility of cross contamination. Before treatment, the Day 20 tissues were transferred from the 'arrival plates' into the wells of the 'treatment plates' containing the maintenance medium.
Control samples:
other: Triton X-100
Amount/concentration applied:
The positive control material, Triton X-100, was prepared as a 0.1% w/v solution in sterile distilled water.
Duration of treatment / exposure:
tissues were treated with the test material for exposure periods of 4 and 24 hours
Duration of post-treatment incubation (if applicable):
three hours incubation at 37"C, 5% CO2 in air.
Number of replicates:
triplicate tissues were treated with the test material
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
at 4h exposure time
Value:
108.1
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
at 24h exposure time
Value:
96.7
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Other effects / acceptance of results:
Assessment of Direct Test Material Reduction of MTT:
The solution did not turn blue/purple. This was taken to indicate that the test material was not able to directly reduce MTT.

Assessment of Skin Irritation Potential:
The relative mean viability of the test material treated tissues was 108.1% after 4 hours exposure and 96.7% after 24 hours exposure. It was considered unnecessary to proceed with tissue histology or analysis of inflammatory mediator levels.

Qualitative Evaluation of Tissue Viability (MTT Uptake Visual Assessment):
Following the 4 and 24 hour exposures the test material treated tissues appeared blue which was considered to be indicative of viable tissue.
Interpretation of results:
GHS criteria not met
Conclusions:
The test material was considered to be a non-irritant under the conditions of this test.
Executive summary:

The purpose of this study was to determine the skin irritation potential of the test material using the SkinEthic Reconstituted Human Epidermal model (RHE, SkinEthic Laboratories, Nice, France) following exposure periods of 4 and 24 hours. The experimental design of the study consists of a test for Direct Reduction of MTT by the test material, followed by the main test. For the main test, triplicate SkinEthic tissues were treated with approximately 25 mg of test material and exposed for 4 hours and 24 hours. The tissues were incubated at 37°C in a humidified atmosphere of 5% C02 in air for the appropriate exposure times. Duplicate untreated tissues were used for each exposure period to serve as negative controls. Duplicate tissues treated with 50 ul of Triton X-100 0.1% w/v were used for the 24 hour exposure period to serve as a positive control. The relative mean viability of the test material treated tissues was 108.1% after 4 hours exposure and 96.7% after 24 hours exposure. It was considered unnecessary to proceed with tissue histology or analysis of inflammatory mediators. In conclusion, the test material was considered to be a non-irritant in this in-vitro study.

Endpoint:
skin irritation: in vivo
Data waiving:
study technically not feasible
Justification for data waiving:
other:
Justification for type of information:
The substance is highly reactive and hydrolyzes in contact with water or body fluids; measured ph-value in water is 2. In literature, the substance is reported to be severly irritant to eyes and is listed listed in Annex VI REACH, therefore it was decided to waive the in-vivo skin irritation study and to conduct an in-vitro test instead.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Referenceopen allclose all

Endpoint:
eye irritation: in vitro / ex vivo
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
the study does not need to be conducted because the substance is classified as skin corrosion, leading to classification as serious eye damage (Category 1)
the study does not need to be conducted because the substance is a strong acid (pH <= 2.0) or base (pH => 11.5) and the available information indicates that it should be classified as serious eye damage (Category 1)
an in vitro eye irritation study does not need to be conducted because adequate data from an in vivo eye irritation study are available
Justification for type of information:
Substance is listed in the consolidated version of EU Regulation 1272/2008 as corrosive, leading to a classification as serious eye damage (Cat. 1). The substance is highly reactive and hydrolyzes in contact with water or body fluids to the corresponding pyromellitic acid; measured ph-value of hydrolyzed material in water is 2. In literature, the substance is reported to be severly irritant to eyes, therefore it was decided to waive the in-vitro eye irritation study
Deviations:
not specified
GLP compliance:
not specified
Specific details on test material used for the study:
not specified
Species:
rabbit
Strain:
not specified
Vehicle:
not specified
Controls:
not specified
Amount / concentration applied:
50 mg
Duration of treatment / exposure:
not specified
Observation period (in vivo):
not specified
Duration of post- treatment incubation (in vitro):
not specified
Number of animals or in vitro replicates:
not specified
Details on study design:
not specified
Endpoint:
eye irritation: in vivo
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
the study does not need to be conducted because the substance is classified as skin corrosion, leading to classification as serious eye damage (Category 1)
Justification for type of information:
Substance is listed in the consolidated version of EU Regulation 1272/2008 as corrosive, leading to a classification as serious eye damage (Cat. 1). The substance is highly reactive and hydrolyzes in contact with water or body fluids to the corresponding pyromellitic acid; measured ph-value of hydrolyzed material in water is 2. In literature, the substance is reported to be severly irritant to eyes, therefore it was decided to waive the in-vivo eye irritation study
Endpoint:
eye irritation: in vivo
Type of information:
other: cross reference to literature data
Adequacy of study:
weight of evidence
Study period:
not specified
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
Standard Draize Test
Deviations:
no
GLP compliance:
not specified
Species:
rabbit
Strain:
not specified
Details on test animals or tissues and environmental conditions:
no data
Vehicle:
unchanged (no vehicle)
Controls:
not specified
Amount / concentration applied:
50 mg
Duration of treatment / exposure:
24 hours in standard draize test
Observation period (in vivo):
24, 48 and 72 hours (up to 21 days) in strandard draize test
Duration of post- treatment incubation (in vitro):
no data
Number of animals or in vitro replicates:
no data
Details on study design:
no data
Irritation parameter:
other: not specified
Basis:
other: not specified
Time point:
other: not specified
Reversibility:
not specified
Remarks on result:
positive indication of irritation
Irritant / corrosive response data:
The substance was observed to cause severe eye irritation, leading to a classification as serious eye damage (Cat. 1).
Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Conclusions:
The substance needs to be considered as severe eye irritant, cat. 1.
Executive summary:

Substance is listed in the consolidated version of EU Regulation 1272/2008 as corrosive, leading to a classification as serious eye damage (Cat. 1).  The substance is highly reactive and hydrolyzes in contact with water or body fluids to the corresponding pyromellitic acid; measured ph-value of hydrolyzed material in water is 2. In literature, the substance is reported to be severly irritant to eyes in a standard Draize test, therefore it was decided to take over this classification and to conduct no further studies.

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

The substance is known as severe eye irritant and listed in Annex VI REACH as corrosive leading to "Causes serious eye damage, Cat. 1" (H 318 - Causes serious eye damage).