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EC number: 228-326-0 | CAS number: 6227-14-1
- 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
Not irritating for skin
Irritating for the eye
Key value for chemical safety assessment
Skin irritation / corrosion
Link to relevant study records
- Endpoint:
- skin corrosion: in vitro / ex vivo
- Type of information:
- other: read across from similar substance
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- study well documented, meets generally accepted scientific principles, acceptable for assessment
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Test system:
- human skin model
- Remarks:
- EPISKIN Reconstructed Human Epidermis Model Kit
- Cell type:
- other: reconstructed (RhE)
- Details on animal used as source of test system:
- The procedure followed is based on the recommended EpiSkin SOP, Version 1.8 (February 2009), ECVAM Skin Irritation Validation Study
- Vehicle:
- unchanged (no vehicle)
- Details on test system:
- A 3 mg/mL MTT stock solution was prepared in DPBS. The stock solution was diluted to 0.3 mg/mL with assay medium when required
- Control samples:
- yes, concurrent negative control
- yes, concurrent positive control
- Number of replicates:
- 3
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- Tested substance - mean of replicates
- Value:
- 117.53
- Vehicle controls validity:
- not examined
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- no indication of irritation
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- Negative control group - mean of replicates
- Value:
- 100
- Vehicle controls validity:
- not specified
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- no indication of irritation
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- Positive control group - mean of replicates
- Value:
- 13.5
- Vehicle controls validity:
- not specified
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- positive indication of irritation
- Interpretation of results:
- GHS criteria not met
- Conclusions:
- Not irritant for skin
- Executive summary:
The substance was tested in the EPISKIN™ Reconstructed Human Epidermis Model using triplicate tissues during 15 minutes. Additional controls were included to account for direct MTT interference and color interference. The relative mean viability of the test substance treated tissues was 59.8 %. The test results does not show any effect on skin. The tested substance could be considered as not irritant for the skin and does not required any classification.
Reference
Cell viability determination is based on cellular mitochondrial dehydrogenase activity, measured by MTT reduction and conversion into blue formazan salt that is quantified after extraction from tissues (Mosman T., 1983). The reduction of cell viability in treated tissues is compared to negative controls and expressed as a percentage. The percentage reduction in viability is used to predict the irritation potential.
Each test substance (test material, negative and positive controls) is topically applied concurrently on three tissues replicates for 42 minutes at room temperature (RT, comprised between 18°C to 24°C). Exposure to the test substance was followed by rinsing with phosphate buffer saline (PBS) and mechanically dried. Epidermis were then transferred to fresh medium and incubated at 37°C for 42 additional hours. Cell viability is assessed by incubating the tissues for 3 hours with 0.3 mL MTT solution (1 mg/mL). The formazan crystals are extracted using 1.5 mL isopropanol for 2 hours at RT and quantified by spectrophotometry at 570 nm wavelength. Sodium Dodecyl Sulphate (SDS 5%), and PBS treated epidermis are used as positive and negative controls, respectively. For each treated tissue, the cell viability is expressed as the percentage of the mean negative control tissues. Values under 50% is qualified the test substance as irritant.
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (not irritating)
Eye irritation
Link to relevant study records
- Endpoint:
- eye irritation, other
- Remarks:
- in vitro
- Type of information:
- other: read across from similar substance
- Adequacy of study:
- key study
- Study period:
- 2017
- Reliability:
- 2 (reliable with restrictions)
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
- Principles of method if other than guideline:
- High opacity scores were obtained in the test and the corneas were stained dark purple by the substance, and thus, the opacity scores may have been due to the staining. Therefore conform the guidance additional histopathology was performed.
- GLP compliance:
- yes (incl. QA statement)
- Species:
- cattle
- Details on test animals or tissues and environmental conditions:
- SOURCE OF COLLECTED EYES
- Source: local abattoir
- Number of animals: not indicated
- Storage, temperature and transport conditions of ocular tissue: placed in Hanks’ Balanced Salt Solution (HBSS) supplemented with antibiotics (penicillin at 100 IU/mL and streptomycin at 100
μg/mL), transported over ice packs on the same day of slaughter. The corneas were prepared immediately on arrival.
- Time interval prior to initiating testing: not indicated
- indication of any existing defects or lesions in ocular tissue samples: checked and only undamaged tissue used - Vehicle:
- physiological saline
- Controls:
- yes, concurrent vehicle
- yes, concurrent positive control
- Amount / concentration applied:
- TEST MATERIAL: 20% w/v solution in sodium chloride 0.9% w/v.
- Duration of treatment / exposure:
- 240 minutes at 32 ± 1 ºC
- Duration of post- treatment incubation (in vitro):
- post treatment opacity measurement immediately
permeality measurement after exposure to fluorescein for 90 min at 32 ± 1 ºC - Details on study design:
- QUALITY CHECK OF THE ISOLATED CORNEAS:based on visual examination and pre-treatment opacity check
NUMBER OF REPLICATES: 2/treatment
TREATMENT METHOD:closed chamber
REMOVAL OF TEST SUBSTANCE: 3 rinses with EMEM containing phenol red
MEASURED ENDPOINTS: Corneal opacity and Corneal permeability
SCORING SYSTEM: In Vitro Irritancy Score (IVIS)
DECISION CRITERIA:
IVIS Classification
≤ 3 No category. Not requiring classification to UN GHS or EU CLP
> 3; ≤55 No prediction of eye irritation can be made
> 55 Category 1. UN GHS or EU CLP Causes serious eye damage
Histologically process and microscopically evaluation of the isolated bovine eye corneas was performed after in vitro testing. - Irritation parameter:
- in vitro irritation score
- Value:
- 138.4
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- positive indication of irritation
- Remarks:
- It can not be excluded that staining of the cornea could have infl uenced the opacity readings
- Irritation parameter:
- histopathological observations
- Value:
- >= 1 - <= 2
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- not determinable
- Remarks:
- indication of irritation is not sufficient to allow the conclusion severely damaging to the eyes
- Other effects / acceptance of results:
- The positive control was slightly out of range. This was evaluated and considered not to have affected the study validity.
The purple staining of cornea by the test substance may have influenced the opacity reading. This was checked by histologically processing and microscopically evaluation of the isolated bovine eye corneas after in vitro testing.
The histopathology findings were only low grade and restricted to the upper layers of the epithelium, so were not typical of the histopathological findings that would be expected following exposure of the cornea to substances having the potential to cause serious eye damage associated with dense corneal opacity (i.e. not consistent with the corneal effects commonly induced by Category 1 substances). So, it is very likely that the high scores for opacity in the BCOP test were due primarily to staining of the cornea by the test item. - Interpretation of results:
- Category 2 (irritating to eyes) based on GHS criteria
- Conclusions:
- The substance is not considered severely damaging to the eyes but it's considered as eye irritant.
- Executive summary:
In a BCOP assay bovine corneas were exposed to the substance and assessed for opacity and permeability. Opacity was strongly increased, while permeability was slightly increased compared to the negative controls. The calculation of the In Vitro Irritancy Score showed that the substance is severely damaging to the eyes. As it could not be excluded that staining of the cornea could have influenced the opacity readings, semi-quantitative histopathological evaluation of the corneas from eyes exposed to the substance was performed. This investigation gave scores below the threshold for classification of this material as Eye Category 1. Therefore the conclusion of the study is that the substance is not considered severely damaging to the eyes.
Reference
Cornea |
Post-Treatment- Pre-Treatment |
Corrected Value
|
OD
|
Corrected Value
|
In Vitro Irritancy Score
|
Negative control |
0 |
|
0.029 |
|
|
|
1 |
|
0.022 |
|
|
|
1 |
|
0.054 |
|
|
Mean |
0.7* |
|
0.035 |
|
1.2 |
Positive Control |
83 |
82.3 |
1.239 |
1.204 |
|
|
84 |
83.3 |
1.609 |
1.574 |
|
|
82 |
81.3 |
1.482 |
1.447 |
|
Mean |
|
82.3 |
|
1.408 |
103.5 |
Test Item |
182 |
181.3 |
0.230 |
0.195 |
|
|
135 |
134.3 |
0.043 |
0.008 |
|
|
94 |
93.3 |
0.243 |
0.208 |
|
Mean |
|
136.3 |
|
0.137 |
138.4 |
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed (irritating)
Respiratory irritation
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
The outcome of the BCOP test based on the histopathology performed is that the substance is not severely irritating to the eyes. Therefore a study according to OECD 492 was initiated which shows that the substance is irritating to the eyes. In a weight of evidence this information will lead to a classification as eye irritant (category 2).
Justification for classification or non-classification
Skin irritation:
The substance was tested in the EpiDerm™ Human Skin Model using duplicate tissues during 3 and 60 minutes. Additional controls were included to account for direct MTT interference and color interference. The relative mean viability of the test substance treated tissues was 102.6 % for the 3 minutes exposure and 96.3% for the 60 minutes exposure. Therefore it can be concluded that the substance is not corrosive to the skin.
The substance was tested in the EPISKIN™
Reconstructed Human Epidermis Model using triplicate tissues during 15
minutes. Additional controls were included to account for direct MTT
interference and color interference. The relative mean viability of the
test substance treated tissues was 59.8 %. Therefore it can be concluded
that the substance is not irritant to the skin.
Eye irritation:
In a BCOP assay bovine corneas were exposed to the substance and assessed for opacity and permeability. Opacity was strongly increased, while permeability was slightly increased compared to the negative controls. The calculation of the In Vitro Irritancy Score showed that the substance is severely damaging to the eyes. As it could not be excluded that staining of the cornea could have influenced the opacity readings, semi-quantitative histopathological evaluation of the corneas from eyes exposed to the substance was performed. This investigation gave scores below the threshold for classification of this material as Eye Category 1. Therefore the conclusion of the study is that the substance is not considered severely damaging to the eyes.
The eye irritation potential of the substance was assessed by means of the Human Cornea Model Test (OECD 492).
The substance proved to be an MTT reducer and interfered with color. Therefore additional controls,viable tissues without MTT addition and freeze-killed tissues were included.The mean relative absorbance value of the test item, corresponding to the cell viability, decreased to 9.7% (threshold for irritancy:≤60%), consequently the test item was irritant to eye and needs to be classified as H319.
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