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EC number: 249-320-4 | CAS number: 28940-11-6
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Endpoint summary
Administrative data
Description of key information
Skin irritation/corrosion: corrosive to the skin (OECD431, GLP, K, rel.1).
Eye damage/irritation: eye damage (classification by default because the substance is corrosive to the skin)
Key value for chemical safety assessment
Skin irritation / corrosion
Link to relevant study records
- Endpoint:
- skin corrosion: in vitro / ex vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 05 - 07 October 2016
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Remarks:
- GLP study conducted according to OECD test Guideline No. 431 without any deviation.
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.40 (In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER))
- Principles of method if other than guideline:
- Not applicable
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- UK GLP Compliance Programme (inspected on 05 July 2016 / signed on 28 October 2016)
- Test system:
- human skin model
- Source species:
- human
- Cell type:
- non-transformed keratinocytes
- Justification for test system used:
- Following the REACH bottom-up strategy, the EpiDerm™ Human Skin Model method was used to assess skin corrosion as recommended in the OECD test guideline No. 431.
- Vehicle:
- unchanged (no vehicle)
- Details on test system:
- RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm™ Reconstructed Human Epidermis Model Kit, MatTek, Bratislava, Slovakia.
- Tissue batch number(s): 23361
- Production Date: 03 October 2016
- Shipping date: 03 October 2016
- Delivery date: 04 October 2016
- Date received: 04 October 2016
- Date of initiation of testing: 06 October 2016
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 3 and 60 minutes at 37 °C in a humidified atmosphere of 5% CO2
- Temperature of post-treatment incubation: 3 h at 37 °C in a humidified atmosphere of 5% CO2
REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps: Rinsing was performed using a wash bottle and was achieved by filling and emptying each tissue approximately 20 times under a constant soft stream of DPBS for approximately 40 seconds to gently remove any residual test item.
- Observable damage in the tissue due to washing: no noted damage to the test item tissue culture surfaces following the rinsing step.
- Modifications to validated SOP: none reported
MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1.0 mg/mL MTT solution
- Incubation time: 3 hours at 37 °C, 5% CO2 in air
- Spectrophotometer: Anthos 2001 microplate reader
- Wavelength: 562 nm
- Filter: not reported
- Filter bandwidth: filter band pass ± 15
- Linear OD range of spectrophotometer: not reported
FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
In order to confirm the integrity of the test system a comparison was made with historical data for negative and positive controls obtained by the laboratory in the previous six months. All values for negative and positive controls were within the historical range and this was taken to indicate the adequate functioning of the test system.
- Viability: OD (540-570 nm) = 1.739 ± 0.134 (acceptance criteria: between 1.0-3.0)
- Barrier function: ET-50 = 6.71 hours (acceptance criteria: between 4.77-8.72 hours)
- Morphology: Tissue viability and the barrier function test are within acceptable ranges and indicate appropriate formation of the epidermal barrier, the presence of a functional stratum corneum, a viable basal cell layer, and intermediate spinous and granular layers.
- Contamination: sterile (Bacteria, yeast and other fungi not detected)
- Reproducibility: For the previous 39 experiments conducted between April 2016 and September 2016 using this test method, the mean OD of the positive control was 0.087 ± 0.022 after 3 minutes of exposure and 0.078 ± 0.023 after 60 minutes of exposure. The mean percentage viability was 4.7 ± 0.9 after 3 minutes of exposure and 4.4 ± 0.8 after 60 minutes of exposure (The assay establishes the acceptance criterion for an acceptable test if the relative mean tissue viability for the positive control treated tissues was ≤15% relative to the negative control treated tissues after 1 hour exposure). In this same period the mean OD of the negative control was 1.922 ± 0.239 after 3 minutes of exposure and 1.866 ± 0.238 after 60 minutes of exposure (The assay establishes the acceptance criterion for an acceptable test if the mean OD for the negative control treated tissues was ≥0.8 and ≤2.8 for every exposure time).
NUMBER OF REPLICATE TISSUES: Duplicate tissues for test item, negative and positive controls
CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- Killed tissues
- Procedure used to prepare the killed tissues : Freeze-killed tissues were prepared prior to the study by placing untreated EPIDERMTM tissues in an empty 12 well plate and storing in a freezer (-14 to -30 °C) for a minimum of 24 hours. Before use each tissue was thawed by placing in 0.9 mL of assay medium for approximately 1 hour at room temperature.
- N. of replicates : 2
- Method of calculation used: True viability = mean OD (treated viable tissues) - [OD (treated killed tissues)-OD (untreated killed tissues)]
NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 1
PREDICTION MODEL / DECISION CRITERIA
- The test substance is considered to be corrosive to skin if the viability after 3 minutes exposure is less than 50%, or if the viability after 3 minutes exposure is greater than or equal to 50 % and the viability after 1 hour exposure is less than 15%.
- The test substance is considered to be non-corrosive to skin if the viability after 3 minutes exposure is greater than or equal to 50% and the viability after 1 hour exposure is greater than or equal to 15%. - Control samples:
- yes, concurrent negative control
- yes, concurrent positive control
- Amount/concentration applied:
- TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25 mg of the unformulated test item (undiluted) wetted with 25 μL of sterile water to increase tissue surface contact.
NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 50 μL of sterile distilled water was used as supplied
POSITIVE CONTROL
- Amount(s) applied (volume or weight): 50 μL of 8.0 N Potassium Hydroxide was used as supplied - Duration of treatment / exposure:
- 3 and 60 minutes.
- Duration of post-treatment incubation (if applicable):
- not applicable
- Number of replicates:
- Duplicate tissues for test item, negative and positive controls
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- 3 minutes exposure
- Value:
- 103.7
- Vehicle controls validity:
- not applicable
- Negative controls validity:
- valid
- Remarks:
- 100%
- Positive controls validity:
- valid
- Remarks:
- 5.5%
- Remarks on result:
- other: No indication of corrosion
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- 60 minutes exposure
- Value:
- 8.2
- Vehicle controls validity:
- not applicable
- Negative controls validity:
- valid
- Remarks:
- 100%
- Positive controls validity:
- valid
- Remarks:
- 5.5%
- Remarks on result:
- other: Positive indication of corrosion
- Other effects / acceptance of results:
- TISSUE VIABILITY:
The relative mean viability of the test item treated tissues was 103.7 and 8.2% after 3 and 60 minutes exposure, respectively.
The relative mean viability of the negative control treated tissues was 100 % after 3 and 60 minutes exposure.
The relative mean viability of the positive control treated tissues was 5.5 after 3 and 60 minutes exposure.
- OTHER EFFECTS:
- Visible damage on test system: none reported
- Direct-MTT reduction: The MTT solution containing the test item turned blue which indicated that the test item directly reduced MTT. Direct reduction was <30% relative to the negative control and therefore acceptable.
- Colour interference with MTT: No
DEMONSTRATION OF TECHNICAL PROFICIENCY: yes
ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The mean OD562 for the negative control treated tissues was 1.709 for the 3 Minute exposure period and 1.742 for the 60 Minute exposure period. The negative control acceptance criteria were therefore satisfied.
- Acceptance criteria met for positive control: The relative mean tissue viability for the positive control treated tissues was 5.5% relative to the negative control following the 60 Minute exposure period. The positive control acceptance criterion was therefore satisfied.
- Acceptance criteria met for variability between replicate measurements: In the range 20 to 100% viability the Coefficient of Variation between the two tissue replicates of each treatment group did not exceed 30%. The acceptance criterion was therefore satisfied.
- Historical data: In order to confirm the integrity of the test system a comparison was made with historical data for negative and positive controls obtained by the laboratory in the previous six months. All values for negative and positive controls were within the historical range and this was taken to indicate the adequate functioning of the test system. - Interpretation of results:
- Category 1B (corrosive) based on GHS criteria
- Remarks:
- combination of category 1B-and-1C
- Conclusions:
- Under the experimental conditions of this study, the test substance is classified as H314 “Causes severe skin burns and eye damage”, combination of category 1B-and-1C, according to EU CLP Regulation (EC) No 1272/2008 and to the GHS.
- Executive summary:
An in vitro skin irritation study was performed according to the OECD Guideline 431 and in compliance with GLP, using the EpiDerm™ Human Skin Model.
Duplicate tissues were treated with 25 mg of the unformulated test item (undiluted) wetted with 25 μL of sterile water to increase tissue surface contact for exposure periods of 3 and 60 minutes. The test item was found to directly reduce MTT. Direct reduction was <30% relative to the negative control and therefore acceptable. Thus additional non-viable tissues were incorporated into the testing for correction purposes. Negative and positive control groups were treated for each exposure period. At the end of the exposure period the test item was rinsed from each tissue before each tissue was taken for MTT‑loading. After MTT loading each tissue was placed in 2 mL Isopropanol for MTT extraction. At the end of the formazan extraction period each well was mixed thoroughly and triplicate 200 mL samples were transferred to the appropriate wells of a pre-labeled 96‑well plate. The optical density (OD) was measured at 562 nm (OD562). Data are presented in the form of percentage viability (MTT reduction in the test item treated tissues relative to negative control tissues).
The relative mean viability of the test item treated tissues was 103.7 and 8.2% after 3 and 60 minutes exposure, respectively. The relative mean viability of the positive control treated tissues was 5.5% after 3 and 60 minutes exposure. The quality criteria required for acceptance of results in the test were satisfied.
Under the experimental conditions of this study, the test substance is classified as H314 “Causes severe skin burns and eye damage”, combination of category 1B-and-1C, according to EU CLP Regulation (EC) No 1272/2008 and to the GHS.
This study is considered as acceptable and satisfies the requirement for skin icorrosion endpoint.
- Endpoint:
- skin irritation: in vitro / ex vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- From 15 December 2015 to 8 February 2016
- 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)
- Version / remarks:
- adopted 28 July 2015
- Deviations:
- no
- Principles of method if other than guideline:
- Not applicable
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- UK GLP Compliance Program (inspected on November 25 to 27, 2015 / Signed on February 15, 2016)
- Test system:
- human skin model
- Source species:
- human
- Cell type:
- non-transformed keratinocytes
- Justification for test system used:
- Following the REACH bottom-up strategy, the EPISKIN™ Reconstructed Human Epidermis Model method was used to assess skin irritation as recommended in the OECD test guideline No. 439.
- Vehicle:
- unchanged (no vehicle)
- Details on test system:
- RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EPISKIN™ Reconstructed Human Epidermis Model Kit, SkinEthic Laboratories, Lyon, France
- Tissue batch number(s): 15 EKIN 050 (test 1) and 16 EKIN 005 (test 2)
- Production date: not reported
- Shipping date: 15 December 2015 (test 1) and 2 February 2016 (test 2)
- Delivery date: 15 December 2015 (test 1) and 2 February 2016 (test 2)
- Expiry date: 23 December 2015 (test 1) and 8 February 2016 (test 2)
- Date of initiation of testing: 15 December 2015 (test 1) and 2 February 2016 (test 2)
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: room temperature
- Temperature of post-treatment incubation (if applicable): 37°C
REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps: At the end of the treatment period, each tissue was rinsed with 25 mL sterile Dulbeccos Phosphate Buffered Saline (DPBS) to remove any residual test or control items and then incubated in 2mL maintenance medium for 42 h at 37 °C, 5 % CO2 in air. Number of washing steps not reported.
- Observable damage in the tissue due to washing: none reported
- Modifications to validated SOP: none reported
MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 0.3 mg/mL
- Incubation time: 3 hours
- Spectrophotometer: BMG Fluostar Optima – plate reader
- Wavelength: 540 nm
- Filter: not reported
- Filter bandwidth: not reported
- Linear OD range of spectrophotometer: not reported
FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: negative control OD values: 0.722, 0.732 and 0.702 (mean historical OD of the negative control was 0.854 ± 0.148)
- Barrier function: IC50 = 2.3 mg/ml (test 1) and 2.1 mg/ml (test 2) ( ≥ 1.5 mg/ml)
- Morphology: Well-differenciated epidermis consisting of a basal layer, several spinous and granular layers and a thinck stratum corneum
- Contamination: absence of bacteria, fungus and mycoplasma
- Reproducibility: For the previous 68 experiments conducted between October 2008 and November 2015 using this test method, the mean OD of the positive control was 0.160 ± 0.074 and the mean percentage viability was 20.8 ± 9.3 (The assay establishes the acceptance criterion for an acceptable test if the relative mean tissue viability for the positive control treated tissues was ≤40% relative to the negative control treated tissues, and the standard deviation value of the percentage viability is ≤18%). In this same period the mean OD of the negative control was 0.782 ± 0.087 (The assay establishes the acceptance criterion for an acceptable test if the mean OD for the negative control treated tissues was ≥0.6 and ≤1.5).
CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE: not applicable
NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 2. The initial test was conducted between 15 December 2015 and 21 December 2015, however, the mean absorbance of the triplicate negative control values was 0.551 which was below the minimum acceptance value of 0.6. The test was repeated between 2 February 2016 and 8 February 2016. The data and results from these two tests are included in the report.
PREDICTION MODEL / DECISION CRITERIA
- The test substance is considered to be irritating to skin if relative mean tissue viability is ≤ 50% after 15 minutes of exposure.
- The test substance is considered to be non-irritating to skin if relative mean tissue viability is > 50% after 15 minutes of exposure. - Control samples:
- yes, concurrent negative control
- yes, concurrent positive control
- Amount/concentration applied:
- TEST MATERIAL
- The test item was used as supplied (undiluted).
- Amount(s) applied (volume or weight with unit): 10 ± 2 mg of the test substance (powder) was dispensed over each tissue. The tissues were wetted with 5 μL of purified water prior to application of the test substance.
NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 10 μL
- Concentration (if solution): DPBS
POSITIVE CONTROL
- Amount(s) applied (volume or weight): 10 μL
- Concentration (if solution): Sodium Dodecyl Sulphate (SDS) at a 5% (w/v) aqueous solution - Duration of treatment / exposure:
- 15 ± 0.5 minutes
- Duration of post-treatment incubation (if applicable):
- 42 hours
- Number of replicates:
- Triplicate tissues for test substance, negative and positive controls
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- 15 minute exposure period and 42 h post-exposure incubation period
- Value:
- 28.6
- Vehicle controls validity:
- not applicable
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- positive indication of irritation
- Other effects / acceptance of results:
- - OTHER EFFECTS:
- Visible damage on test system: none reported
- Direct-MTT reduction: no
- Colour interference with MTT: no
DEMONSTRATION OF TECHNICAL PROFICIENCY: yes
ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The mean absorbance of the triplicate negative control values for the original test (test 1) was 0.551 which was below the minimum acceptance value of 0.6. The mean absorbance for the negative control in the repeat test (test 2) was 0.746 which was between the minimum and maximum values of 0.6 and 1.5. The standard deviation (SD) of the % viability for test 1 and 2 was 8.3 and 6.1 respectively. Both test values were below the maximum value of 18. The negative control acceptance criteria were therefore met for the test 2.
- Acceptance criteria met for positive control: The percentage mean viability of the positive control for test 1 was 17.3 ± 0.3 and test 2 was 10.3 ± 3.7 of the negative control. These were below the maximum acceptance values of 40% viability and SD of 18%. The positive control acceptance criteria were therefore met.
- Acceptance criteria met for variability between replicate measurements: Skin irritation is expressed as the remaining cell viability after exposure to the test substance. The relative mean tissue viability obtained after 15 minutes treatment with test item compared to the negative control tissues was 28.6% in test 2. The standard deviation calculated from individual tissue viabilities of the three identically test substance treated tissues were 18.2%, which was above the acceptance criterion of 18.0. In test 1, the standard deviation calculated from individual tissue viabilities was 25.9. The results from test 2 are consistent with the first test in that both % viabilities are less than 50% and the standard deviations are greater than 18, which may be possibly due to test material sticking to and/or penetrating the Episkin tissues. As the standard deviation for the repeat test was only 0.2 greater than 18.0 and that all other acceptance criteria were met the repeat test was considered valid.
- Range of historical values if different from the ones specified in the test guideline: For the previous 68 experiments conducted between October 2008 and November 2015 using this test method, the mean OD of the positive control was 0.160 ± 0.074 and the mean percentage viability was 20.8 ± 9.3. In this same period the mean OD of the negative control was 0.782 ± 0.087. - Other effects:
- None
- Interpretation of results:
- Category 2 (irritant) based on GHS criteria
- Conclusions:
- Under the experimental conditions of this study, the test substance is classified as H315 “Causes Skin Irritation” Category 2 according to Regulation (EC) No. 1272/2008 (CLP) and to the GHS.
- Executive summary:
An in vitro skin irritation study was performed according to the OECD Guideline 439 and in compliance with GLP, using the EPISKINTM reconstructed human epidermis model.
Triplicate tissues were treated with 10 ± 2 mg of the test substance for an exposure period of 15 ± 0.5 minutes. The tissues were wetted with 5 μL of purified water prior to application of the test substance. At the end of the exposure period each tissue was rinsed before incubating for 42 h. At the end of the post-exposure incubation period each tissue was taken for MTT-loading. After MTT-loading a biopsy of each epidermis was made and the tissues were placed into acidified isopropanol for extraction of formazan crystals out of the MTT-loaded tissues. At the end of the formazan extraction period, duplicate 200 μL samples were transferred to the wells of 96-well plate and the optical density was measured at 540 nm.
The test substance was tested on two separate occasions. In the initial test (test 1), the mean absorbance of the triplicate negative control values was 0.551 which was below the minimum acceptance value of 0.6. As test 1 was considered invalid the test was repeated. In the repeat test (test 2), negative and positive controls results were within the acceptable range and the test was considered valid and the quality criteria required for acceptance of results in the test were satisfied. The conclusion was based on the results from the repeat test (test 2).
The test substance elicited a mean tissue viability of 28.6 ± 18.2% in test 2. The standard deviation calculated from individual tissue viabilities of the three identically test substance treated tissues were 18.2%, which was above the acceptance criterion of 18.0. In test 1, the standard deviation calculated from individual tissue viabilities was 25.9. The results from test 2 are consistent with the first test in that both % viabilities are less than 50% and the standard deviations are greater than 18, which may be possibly due to test material sticking to and/or penetrating the Episkin tissues. As the standard deviation for the repeat test was only 0.2 greater than 18.0 and that all other acceptance criteria were met the repeat test was considered valid.
Under the experimental conditions of this study, the test substance is classified as H315 “Causes Skin Irritation” Category 2 according to Regulation (EC) No. 1272/2008 (CLP) and to the GHS.
This study is considered as acceptable and satisfies the requirement for skin irritation endpoint.
Referenceopen allclose all
Table 7.3.1/1: Mean OD562 Values and Viabilities for the Negative Control Item, Positive Control Item and Test Item
Tissue |
Exposure Period |
Mean OD562 of individual tissues |
Mean OD562of duplicate tissues (tvt) |
Corrected OD562 of tissues |
Standard Deviation |
Coefficient of Variation (%) |
Relative Mean Viability (%) |
Negative Control |
3 Minutes |
1.625 |
1.709 |
- |
0.119 |
7.0 |
100* |
1.793 |
|||||||
60 Minutes |
1.774 |
1.742 |
- |
0.046 |
2.6 |
||
1.709 |
|||||||
Positive Control |
3 Minutes |
0.099 |
0.094 |
- |
0.008 |
N/A |
5.5 |
0.088 |
|||||||
60 Minutes |
0.099 |
0.096 |
- |
0.005 |
N/A |
5.5 |
|
0.092 |
|||||||
Test Item |
3 Minutes |
1.733 |
1.776 |
1.772 |
0.060 |
3.4 |
103.7 |
1.818 |
|||||||
60 Minutes |
0.254 |
0.269 |
0.144 |
0.021 |
7.6 |
8.2 |
|
0.283 |
tvt = treated viable tissues
tkt = treated killed tissues
ukt = untreated killed tissues
3 minute exposure corrected mean OD562:
x̅ (0.157 + 0.156) = 0.157 (tkt) – x̅ (0.159 + 0.147) = 0.153 (ukt) = 0.004
60 minute exposure corrected mean OD562:
x̅ (0.319 + 0.255) = 0.287(tkt) – x̅ (0.174 + 0.149) = 0.162 (ukt) = 0.125
3
minute exposure direct MTT reduction relative to the negative control =
0.2%
60 minute exposure direct MTT reduction relative to the negative control
= 7.2%
* = The mean % viability of the negative control tissue is set at 100%
Table 7.3.1/1: EpiSkin™ results
Test |
Sample |
Tissue viability as % of mean OD negative control |
Prediction MTT endpoint |
|||
Replicate Tissues |
Mean ± SD |
|||||
a |
b |
c |
||||
Test 1 |
Test substance |
39.0 |
9.6 |
61.3 |
36.7 ± 25.9 |
Category 2 |
Negative control |
97.6 |
93.2 |
109.20 |
100.0 ± 8.3 |
Not applicable |
|
Positive control |
17.4 |
17.4 |
16.9 |
17.3 ± 0.3 |
Category 2 |
|
Test 2 |
Test substance |
39.1 |
39.0 |
7.6 |
28.6 ± 18.2 |
Category 2 |
Negative control |
99.7 |
94.0 |
106.3 |
100.0 ± 6.1 |
Not applicable |
|
Positive control |
13.9 |
6.5 |
10.5 |
10.3 ± 3.7 |
Category 2 |
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed (corrosive)
Eye irritation
Link to relevant study records
- Endpoint:
- eye irritation: in vitro / ex vivo
- Data waiving:
- study scientifically not necessary / other information available
- Justification for data waiving:
- the study need not be conducted because the available information indicates that the criteria are met for classification as corrosive to the skin or irritating to eyes
- Justification for type of information:
- JUSTIFICATION FOR DATA WAIVING
In accordance with REACH Regulation (EC) 1907/2006 as amended: Annex VII, section 8.2: the serious eye damage / eye irritation study does not need to be conducted if the substance is classified as skin corrosion, leading to classificiation as serious eye damage (Category 1). The substance was shown to be corrosive to the skin in a new in vitro skin corrosion EpiDerm test (Envigo, 2017, Rel.1), therefore the substance was considered to be able to induce serious eye damage and no further in vitro test was needed. - Reason / purpose for cross-reference:
- data waiving: supporting information
Reference
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed (irreversible damage)
Respiratory irritation
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed (irritating)
Additional information
Skin irritation/corrosion:
Since no key study was identified on the registered substance, the testing and assessment strategy, as described in ECHA R.7a Endpoint specific guidance (December 2016), was used to evaluate the skin corrosion/irritation potential of the registered substance:
|
Element |
Information |
Conclusion |
Comments |
Existing data on physico |
1a |
Is the substance spontaneously flammable in contact with air (pyrophoric) or water at room temperature? |
NO |
|
1b |
Is the substance an organic hydroperoxide or an organic peroxide? |
NO |
|
|
1c |
Is the pH of the substance ≤ 2.0 or ≥ 11.5? |
NO |
|
|
1d |
Are there other physical or chemical properties that |
NO |
|
|
Existing human data |
2 |
Are there adequate existing human data which provide evidence that the substance is a corrosive |
NO |
|
Existing animal data from corrosion/irritation studies |
3 |
Are there data from existing studies on corrosion and irritation in laboratory animals, which provide sound conclusive evidence that the substance is a corrosive, irritant or non-irritant? |
NO |
|
Existing data from general toxicity studies via the dermal route and from sensitisation studies |
4a |
Is the substance classified as fatal in contact with skin (LD50 ≤ 50 mg/kg bw, CLP hazard statement |
NO |
|
4b |
Has the substance proven to be a corrosive, irritant or non-irritant in a suitable acute dermal toxicity test? |
NO |
|
|
4c |
Has the substance proven to be a corrosive or an irritant in sensitisation studies or after repeated |
NO |
LLNA available: tested only up to 30%. No sign of irritation or corrosion leading ot C&L. |
|
Existing/new (Q)SAR data and read |
5a |
Are there structurally related substances (suitable “read-across” or grouping), which are classified as corrosive to the skin (Skin Corrosive Cat. 1), or do suitable (Q)SAR methods indicate corrosion |
NO |
|
5b |
Are there structurally related substances (suitable “read-across” or grouping), which are classified as irritant to the skin (Skin Irritant Cat. 2), or indicating that the substance is non-irritant, or do suitable (Q)SAR methods indicate irritant or non-irritant potential of the substance? |
NO |
|
|
Existing in vitro data |
6a |
Has the substance demonstrated corrosive properties in an EU/OECD adopted in vitro test? |
NO |
|
6b |
Has the substance demonstrated irritant or non-irritant properties in an EU/OECD adopted |
NO |
(at the initiation of the dossier, no test was available) |
|
6c |
Are there data from a non-validated suitable in vitro test(s), which provide sound conclusive evidence that the substance is corrosive/ irritant? |
NO |
|
|
Weight-of- Evidence analysis |
7 |
The “elements” described above may be arranged as appropriate. Taking all available existing and |
NO |
|
New in vitro test for irritation |
8 |
Does the substance demonstrate irritating or non-irritating properties in (an) EU/OECD adopted in vitro test(s) for skin irritation? |
YES |
=> an Episkin test for irritation was initiated (Bottom-up strategy - substance expected to be non corrosive). The conclusion of this Episkin test is not sufficient to conclude on C&L, a skin corrosion test was performed to assess the potential corrosivity of the substance to the skin. |
New in vitro test for corrosivity |
9 |
Does the substance demonstrate corrosive properties in (an) EU/OECD adopted in vitro test(s) for skin corrosion? |
YES |
=> an EpiDerm test for corrosion was initiated (Bottom-up strategy - substance expected to be non corrosive). The conclusion of this EpiDerm test is sufficient to conclude on C&L (3 minutes = 103.7%; 60 minutes = 8.2% => corrosive category 1B/1C) |
New in vivo test for corrosion/irritation |
10 |
To be used only as a last resort |
NO |
In vivo testing should not be conducted in this case since the substance falls under the scope of the specific in vitro tests performed, and there are no substance-specific limitations on use of those tests. An adaptation according to Annex XI to the REACH Regulation is included in this dossier. |
Following the REACH bottom-up strategy, an in vitro skin irritation study (Envigo, 2016, Rel.1) was performed according to the OECD Guideline 439 and in compliance with GLP, using the EPISKIN reconstructed human epidermis model. The test substance was tested on two separate occasions. In the initial test (test 1), the mean absorbance of the triplicate negative control values was 0.551 which was below the minimum acceptance value of 0.6. As test 1 was considered invalid the test was repeated. In the repeat test (test 2), negative and positive controls results were within the acceptable range and the test was considered valid. The standard deviation calculated from individual tissue viabilities of the three identically test substance treated tissues were 18.2%, which was above the acceptance criterion of 18.0. In test 1, the standard deviation calculated from individual tissue viabilities was 25.9. The results from test 2 are consistent with the first test in that both % viabilities are less than 50% and the standard deviations are greater than 18, which may be possibly due to test material sticking to and/or penetrating the Episkin tissues. As the standard deviation for the repeat test was only 0.2 greater than 18.0 and that all other acceptance criteria were met the repeat test was considered valid. The conclusion was based on the results from the repeat test (test 2). The relative mean viability of the test item treated tissues was 28.6 %, after the 15‑minute exposure period. With a tissue viability < 50%, the test material was considered to be irritant to skin.
In compliance with bottom-up strategy and to determine the potential of the substance to induce skin corrosion, an in vitro skin corrosion study (Envigo, 2017, Rel.1) was performed according to the OECD Guideline 431 and in compliance with GLP, using the EpiDerm™ Human Skin Model. The relative mean viability of the test item treated tissues was 103.7 and 8.2% after 3 and 60 minutes exposure, respectively. The relative mean viability of the positive control treated tissues was 5.5% after 3 and 60 minutes exposure. The quality criteria required for acceptance of results in the test were satisfied. With a tissue viability < 15% after 60 minutes of exposure, the test material was considered to be corrosive to skin.
Eye irritation:
Since no key study was identified on the registered substance, the testing and assessment strategy, as described in ECHA R.7a Endpoint specific guidance (December 2016), was used to evaluate the eye damage/irritation potential of the registered substance:
|
Element |
Information |
Conclusion |
Comments |
Conclusion of the information strategy on skin corrosion/irritation |
0 |
Is the substance classified as a skin corrosive? |
NO |
|
Existing data on physico |
1a |
Is the substance spontaneously flammable in contact with air (pyrophoric) or water at room temperature? |
NO |
|
1b |
Is the substance an organic hydroperoxide or an organic peroxide? |
NO |
|
|
1c |
Is the pH of the substance ≤ 2.0 or ≥ 11.5? |
NO |
|
|
1d |
Are there other physical or chemical properties that indicate that the substance causes serious eye damage or eye irritation? |
NO |
|
|
Existing human data |
2 |
Are there adequate existing human data which provide evidence that the substance has the potential to cause serious eye damage or eye irritation? |
NO |
|
Existing animal data from corrosion/irritation studies |
3 |
Are there data from existing studies on corrosion and irritation in laboratory animals, which provide sound conclusive evidence that the substance is a corrosive, irritant or non-irritant? |
NO |
|
Existing/new (Q)SAR data and read-across |
4 |
Are there structurally related substances (suitable “read-across” or grouping), which are classified as causing serious eye damage/eye irritation, or indicating that the substance is non-irritant, or do valid (Q)SAR methods indicate serious eye damage/eye irritation or non-irritation of the substance? |
NO |
|
Existing in vitro data |
5a |
Has the substance demonstrated serious eye damage, eye irritation or non-irritating properties in an EU/OECD adopted in vitro test? |
NO |
(at the initiation of the dossier, no test was available) |
5b |
Are there acceptable data from (a) non-validated suitable in vitro test(s), which provide sound evidence that the substance causes serious eye damage/eye irritation? |
NO |
|
|
Weight-of- Evidence analysis |
6 |
The “elements” described above may be arranged as appropriate. Taking all available existing and relevant data mentioned above (Elements 0 – 5) into account, is there sufficient information to make a decision on whether classification/labelling is necessary, and – if so – how to classify and label? |
NO |
|
New in vitro tests for serious eye damage/eye irritation (Annex VII to the REACH Regulation) |
7a |
Does the substance demonstrate serious eye damage, eye irritation or non-irritant properties in (an) EU/OECD adopted in vitro test(s) for the eye hazard charaterisation? |
YES |
The substance was shown to be corrosive to the skin in a new in vitro skin corrosion EpiDerm test |
8b |
Does the substance demonstrate serious eye damage or eye irritant properties in (a) non-validated suitable in vitro test(s) for serious eye damage/eye irritation? |
NO |
|
|
New in vivo test for serious eye damage/eye irritation as a last resort (Annex VIII to the REACH Regulation) |
8b |
Does the substance demonstrate serious eye damage or eye irritation in an OECD adopted in vivo test? |
NO |
In vivo testing should not be conducted in this case since the substance falls under the scope of the specific in vitro tests performed, and there are no substance-specific limitations on use of those tests. An adaptation according to Annex XI to the REACH Regulation is included in this dossier. |
Since the substance was shown to be corrosive to the skin in the new in vitro skin corrosion EpiDerm test, therefore the substance was considered to be able to induce serious eye damage and no further in vitro test was needed as the conclusion of this skin corrosion test is sufficient to conclude on C&L.
Justification for classification or non-classification
Harmonized classification:
The substance has no harmonized classification according to the Regulation (EC) No. 1272/2008.
Self-classification:
Based on the available information, the substance should be classified as skin corrosive sub-category 1B (H314 “Causes severe skin burns and eye damage) according to the Annex VI of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.
Therefore, the substance is also classified for serious eye damage Category 1 according to the Annex VI of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.
No data was available regarding respiratory irritation. In general, a classification for corrosivity is considered to implicitly cover the potential to cause respiratory tract irritation and so the additional Category 3 is considered to be superfluous. However, such substance has to be supplementary labelled with EUH071 if there is a possibility of exposure via inhalation taking into consideration the saturated vapour concentration and the possibility of exposure to particles or droplets of inhalable size. It is also strongly recommended to apply the precautionary statement P260: "Do not breathe dust/fume/gas/mist/vapours/spray."
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.