Registration Dossier
Registration Dossier
Data platform availability banner - registered substances factsheets
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Diss Factsheets
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 247-196-6 | CAS number: 25707-70-4
- 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
The in vitro skin corrosion test in EpiDermTM model (OECD 431) indicates that the test item is corrosive.
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:
- 2021-11-10 to 2021-11-10
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
- Version / remarks:
- 2019-06-18
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.40 (In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER))
- Version / remarks:
- amended 2019-07-31
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Test system:
- human skin model
- Source species:
- human
- Cell type:
- non-transformed keratinocytes
- Cell source:
- other: not specified
- Source strain:
- other: human adult donor
- Justification for test system used:
- The corrosivity potential of a chemical may be predicted by measurement of its cytotoxic effect, as reflected in the MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, Thiazolyl blue; CAS number 298-93-1] assay, on the EpiDerm™ reconstituted human epidermis. This method is approved by international regulatory agencies as a replacement for the identification of corrosives in the in vivo Rabbit skin assay (OECD 404) and is specifically approved as a replacement for the in vivo skin corrosivity test within OECD 431.
- Details on test system:
- RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: The reconstructed human epidermal model EpiDerm™ (EPI-200, MatTek, Ashland, MA, USA and Bratislava, Slovakia)
- Tissue batch number: 36103
- Shipping date: 08 November 2021
- Delivery date: 09 November 2021
- Date of initiation of testing: 12 November 2021
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during exposure: 37±1 °C
- Temperature of post-treatment incubation: 37±1 °C
REMOVAL OF TEST MATERIAL AND CONTROLS
- Washing steps:
After the exposure time the EpiDerm™ units were removed and rinsed thoroughly with 1x DPBS sterile solution to remove all of the test material from the epidermal surface. The constant soft stream of DPBS was used from approximately 1.5 cm distance, and filling and emptying the tissue insert was repeated 20 times. The rest of the DPBS was decanted onto the absorbent material. Decanting was most efficiently performed by rotating the insert to approximately a 45° angle and touching the upper lip to the absorbent material (to break the surface tension).
Following rinsing, the tissues were transferred and kept in the 24-well holding plates (pre-filled with 300 μL medium) until the rinsing procedure was completed. The rest of the DPBS was removed from the tissues by gently sweeping the tissues’ surface with a sterile cotton tipped swab (care was taken to avoid the damage of tissues).
- Observable damage in the tissue due to washing: no
- Modifications to validated SOP: none
MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL MTT per well
- Incubation time: 3 hours (± 5 min)
- Spectrophotometer: Varioskan™ LUX Type 3020
- Wavelength: 570 nm
- Linear OD range of spectrophotometer: 200 – 1000 nm
FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
The EpiDerm™ System is manufactured according to defined quality assurance procedures. All biological components of the epidermis and the culture medium are tested by manufacturer for viral, bacterial, fungal and mycoplasma contamination. MatTek determines the ET-50 value following exposure to Triton X-100 (1%) for each EpiDerm™ lot. The ET-50 must fall within a range established based on a historical database of results.
NUMBER OF REPLICATE TISSUES: 3
CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- Killed tissues: for the MTT evaluation, batch No. of killed epidermis: 34139
- N. of replicates: 3
- Method of calculation used: See "Any other information on materials and methods, incl. tables"
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 is < 50 % after 3 min exposure OR ≥ 50 % after 3 min exposure AND < 15 % after 60 min exposure.
- The test substance is considered to be non-corrosive to skin if the viability is ≥ 50 % after 3 min exposure AND ≥ 15 % after 60 min exposure
If the test substance is considered to be corrosive, then the following criteria apply:
- If the cell viability is < 25 % after 3 min exposure, then the the test substance is clasified as optional Sub-category 1A
- If the cell viability is ≥ 25 % after 3 min exposure, then the the test substance is clasified as a combination of optional Sub-categories 1B-and-1C - Control samples:
- yes, concurrent vehicle
- yes, concurrent positive control
- Amount/concentration applied:
- 50 µL
- Duration of treatment / exposure:
- 60 and 3 minutes
- Duration of post-treatment incubation (if applicable):
- MTT assay: 3 hours (± 5 min);
The exposure of test item was terminated by rinsing with 1xDPBS, the EpiDerm™ units were transferred into the MTT ready to use solution filled 24-well plate (300 μL of 1 mg/mL MTT per well) and then incubated at 37±1 °C in an incubator with 5±1 % CO2, ≥95 % humidified atmosphere. - Number of replicates:
- for both exposure times: 3 replicates of the test item, 3 replicates of negative control and 3 replicates of positive control were used. Furthermore, 3 killed test item-treated tissues and 3 killed negative control-treated tissues were used for the MTT evaluation for each exposure time.
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- 1 / 3 min
- Value:
- 64
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- positive indication of irritation
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- 1/ 60 min
- Value:
- 10
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- positive indication of irritation
- Other effects / acceptance of results:
- The mean OD value of the three negative control tissues was 2.038 and 2.140 at 60 and 3 minutes exposure respectively. The mean OD value obtained for the positive control was 0.104 at
60 minutes exposure which result corresponds to 5 % viability, when compared to the results obtained from the negative control. Each calculated Coefficient of Variation value (CV) in the range of 20-100 % viability was below 30. The mean OD value of the blank samples MTT-100-EXT were 0.0375 and 0.0376 (below 0.1).
All validity criteria were within acceptable limits and therefore the study was considered to be valid.
The test item interacted with the MTT, therefore additional controls and data calculations were necessary.
The mean non-specific MTT reduction (NSMTT) was determined to be 2 % and 0 % at the 60 min and 3 min exposure respectively. As the NSMTT were below 30 % in each case. However, the true MTT metabolic conversion was made and the correction of viability percentages were undertaken after the 60 minutes exposure, because the correction with 0% was not interpretable at the 3 minutes exposure. - Interpretation of results:
- Category 1B (corrosive) based on GHS criteria
- Conclusions:
- In conclusion, in this in vitro skin corrosion test in EpiDermTM model (OECD 431) with the test sbstance, the results indicate that the test item is corrosive. According to the UN GHS classification systems, the test sbstance has been categorized as “Corrosive: A combination of optional sub-categories 1B and 1C”.
- Executive summary:
EpiDermTM test of the test item has been performed to predict its corrosion potential by measurement of its cytotoxic effect, as reflected in the MTT assay according to the OECD Test Guideline No. 431, 18 June 2019. Disks of EpiDermTM (three units / chemical / incubation time) were treated with the test item and incubated for 60 minutes (+ 3 min) at standard culture condition (37±1 °C in an incubator with 5±1 % CO2 in a 95 % humidified atmosphere) and 3 min at room temperature. Exposure of test material was terminated by rinsing with 1x DPBS solution. The viability of each disk was assessed by incubating the tissues for 3 hours (±5 min) with MTT solution at 37±1 °C in an incubator with 5±1 % CO2 in a 95 % humidified atmosphere and protected from light. The formazan precipitated was then extracted using MTT-100-EXT and quantified spectrophotometrically. Potassium hydroxide (KOH) 8N solution and sterile deionized water treated (three units / positive and negative control) epidermis were used as positive and negative controls, respectively. For each treated tissue, viability was expressed as a percentage relative to the negative control.
The test item is a MTT-reducer, therefore additional controls (test item treated killed tissues and negative control treated killed tissues) were used to detect and correct for test item interference with the viability measurement.
For each treated tissue, viability was expressed as % relative to the negative control. The test item is considered to be non-corrosive to skin, if the mean relative viability after 60 minutes of exposure is above or equal 15 % and the mean relative viability after 3 minutes of exposure is above or equal 50 % of the negative control.
The test item showed significantly reduced cell viability in comparison to the negative control after 60 minutes of exposure. However, the test item did not show significantly reduced cell viability in comparison to the negative control after 3 minutes of exposure. The average test item treated tissue relative viability was 10 % at 60 minutes of exposure and the mean viability was 64 % at 3 minutes of exposure.
Positive and negative controls showed the expected optical density (OD) and cell viability values within acceptable limits. All assay acceptance criteria were met, the experiment was considered to be valid.
In conclusion, in this in vitro skin corrosion test in EpiDermTM model (OECD 431) with ANCAMINE 2458, the results indicate that the test item is corrosive. According to the UN GHS classification systems, the test sbstance has been categorized as “Corrosive: Optional Sub- categories 1B and 1C”.
Reference
OD values and viability percentages of the controls:
Controls | Optical Density (OD) | Viability (%) | |||
Negative Control: | 1 | 2.139 | 105 | ||
2 | 1.996 | 98 | |||
3 | 1.980 | 97 | |||
mean | 2.038 | 100 | |||
SD |
| 4.303 | |||
CV |
| 4.303 | |||
Negative Control: | 1 | 2.100 | 98 | ||
2 | 2.319 | 108 | |||
3 | 2.000 | 93 | |||
mean | 2.140 | 100 | |||
SD |
| 7.630 | |||
CV |
| 7.630 | |||
Positive Control: | 1 | 0.129 | 6 | ||
2 | 0.068 | 3 | |||
3 | 0.114 | 6 | |||
mean | 0.104 | 5 | |||
SD |
| 1.560 | |||
CV |
| 30.607 | |||
Positive Control: | 1 | 0.282 | 13 | ||
2 | 0.546 | 26 | |||
3 | 0.325 | 15 | |||
mean | 0.384 | 18 | |||
SD |
| 6.627 | |||
CV |
| 36.902 |
OD values and viability percentages of the test item (including corrected values):
Substance | Optical Density (OD) | TODTT | Viability (%) | Relative Viability (%) | |
Test Item: 60 min exposure | 1 | 0.216 | 0.185 | 11 | 9 |
2 | 0.191 | 0.160 | 9 | 8 | |
3 | 0.322 | 0.291 | 16 | 14 | |
mean | 0.243 | 0.212 | 12 | 10 | |
SD |
| 3.413 | 3.413 | ||
CV |
| 28.654 | 32.843 |
OD values and viability percentages of the test item:
Test Item | Optical Density (OD) | Viability (%) | |
Test Item: 3 min exposure | 1 | 1.486 | 69 |
2 | 1.239 | 58 | |
3 | 1.383 | 65 | |
mean | 1.369 | 64 | |
SD |
| 5.805 | |
CV |
| 9.072 |
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed (corrosive)
Eye irritation
Endpoint conclusion
- Endpoint conclusion:
- no study available
Respiratory irritation
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
in vitro corrosion
EpiDermTM test of the test item has been performed to predict its corrosion potential by measurement of its cytotoxic effect, as reflected in the MTT assay according to the OECD Test Guideline No. 431, 18 June 2019. Disks of EpiDermTM (three units / chemical / incubation time) were treated with the test item and incubated for 60 minutes (+ 3 min) at standard culture condition (37±1 °C in an incubator with 5±1 % CO2 in a 95 % humidified atmosphere) and 3 min at room temperature. Exposure of test material was terminated by rinsing with 1x DPBS solution. The viability of each disk was assessed by incubating the tissues for 3 hours (±5 min) with MTT solution at 37±1 °C in an incubator with 5±1 % CO2 in a 95 % humidified atmosphere and protected from light. The formazan precipitated was then extracted using MTT-100-EXT and quantified spectrophotometrically. Potassium hydroxide (KOH) 8N solution and sterile deionized water treated (three units / positive and negative control) epidermis were used as positive and negative controls, respectively. For each treated tissue, viability was expressed as a percentage relative to the negative control.
The test item is a MTT-reducer, therefore additional controls (test item treated killed tissues and negative control treated killed tissues) were used to detect and correct for test item interference with the viability measurement.
For each treated tissue, viability was expressed as % relative to the negative control. The test item is considered to be non-corrosive to skin, if the mean relative viability after 60 minutes of exposure is above or equal 15 % and the mean relative viability after 3 minutes of exposure is above or equal 50 % of the negative control.
The test item showed significantly reduced cell viability in comparison to the negative control after 60 minutes of exposure. However, the test item did not show significantly reduced cell viability in comparison to the negative control after 3 minutes of exposure. The average test item treated tissue relative viability was 10 % at 60 minutes of exposure and the mean viability was 64 % at 3 minutes of exposure.
Positive and negative controls showed the expected optical density (OD) and cell viability values within acceptable limits. All assay acceptance criteria were met, the experiment was considered to be valid.
In conclusion, in this in vitro skin corrosion test in EpiDermTM model (OECD 431), the results indicate that the test item is corrosive. According to the UN GHS classification systems, the test item has been categorized as “Corrosive: Optional Sub- categories 1B and 1C”.
Justification for classification or non-classification
Classification, Labelling, and Packaging Regulation (EC) No 1272/2008
The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008.
Skin:
Based on available data the test item is corrosive to skin. Based on the conducted test, the test substance requires classification as “Corrosive: A combination of optional sub-categories 1B and 1C” according to Regulation (EC) No 1272/2008 (CLP), as amended for the seventeenth time in Regulation (EU) No 2021/849. In consideration of the worst-case, the test substance is subcategorizd to Category 1B.
Eye:
Additionaly, the test item requires classification as serious eye damage (Category 1) according to Regulation (EC) No 1272/2008 (CLP), as amended for the seventeenth time in Regulation (EU) No 2021/849.
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.