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EC number: 268-211-2 | CAS number: 68037-36-5
- 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
Skin irritation / corrosion
Administrative data
- Endpoint:
- skin corrosion: in vitro / ex vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- experimental part of study performed in period from 2013-09-11 to 2013-09-16
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Guideline study from supporting substance (structural analogue)
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 013
- Report date:
- 2013
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.40 (In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER))
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
Test material
- Test material form:
- solid: particulate/powder
Constituent 1
In vitro test system
- Test system:
- human skin model
- Source species:
- human
- Cell type:
- non-transformed keratinocytes
- Vehicle:
- water
- Details on test system:
- Test system
The reconstructed human epidermal model EpiDerm (EPI-200, MatTek, Ashland, USA) which consist of normal human derived epidermal keratinocytes cultured to form a multilayered highly differentiated model of the human epidermis. the EpiDerm tissues (area 0.63 cm2) are cultured on specially prepared cell culture inserts and shipped as kits.
Test procedure
a) Test substance application
The test substance (0.25 mg) was placed directly atop to the previously moistened tissue with 25 μL water. The material was spread on the tissue surface.
Procedure
On the day of experiment, EpiDerm tissues were conditioned by incubation to release transport stress related compounds and debris. After pre-incubation, tissues were topically exposed to the test chemical for 3 and 60 minutes. In each time interval three tissues were used per test chemical, three for the positive control (PC) and three for negative control (NC). After exposition, tissues were thoroughly rinsed and blotted to remove the test substance (controls).
After that, tissues were transferred to 24-well plates containing MTT medium (1 mg/mL). After 3 hour MTT incubation, the blue formazan salt formed by cellular mitochondria was extracted with 2.0 mL/tissue of isopropylalcohol and the optical density of the extracted formazan was determined using spectrophotometer Libra S22 at 570 nm. Isopropylalcohol serves as a blank. - Control samples:
- yes, concurrent negative control
- yes, concurrent positive control
- Amount/concentration applied:
- 0.25 mg was placed directly atop to the previously moistened tissue with 25 μL water
- Duration of treatment / exposure:
- After pre-incubation, tissues were topically exposed to the test chemical for 3 and 60 minutes.
- Number of replicates:
- 2
Test animals
- Details on test animals or test system and environmental conditions:
- in vitro test on reconstructed human epidermis
Test system
- Amount / concentration applied:
- see Any other information on materials and methods incl. tables
Results and discussion
In vitro
Resultsopen allclose all
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- 3 min run
- Value:
- 109.5
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- no indication of irritation
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- 60 minute treatment
- Value:
- 76.5
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- no indication of irritation
- Other effects / acceptance of results:
- The testing of possible interference of test substance with test endpoint was performed. Although, after direct reduction in test-tube, there were suspicion, that the test substance was directly reducing, test in frozen tissues did not confirm this assumption. Then, the correction of results was not necessary.
As it is possible to see from the results given in Table 2, average viability of affected tissues was 109.5 % of negative control average value after 3 min treatment and 76.5 % after 60 min.
According to evaluation criteria given in Chapter 3.7 both the two values were higher than critical values (50 % and 15% respectively): 109.5 % ≥ 50 % after 3 min and 76.5 % ≥ 15 % after 60 min. The test substance should be regarded as non-corrosive.
Any other information on results incl. tables
1.1. Direct MTT reduction- functional check in tubes
50 mgof the test substance was added to 2 mL of MTT medium. Solution was incubated for 1 hour (37±1°C, 5±1 % CO2, moistened).
The test substance was filtered because of betterevaluation.The test substance changed colour from green to blue (see Figure 1)
Next step – test in frozen tissues - had to be done for confirmation/excluding of a direct reduction in tissues.
1.2. Direct MTT reduction - test in frozen tissues
The test substance (25 mg) was applied to two freeze-killed tissues for 3 min exposition and two freeze-killed tissues for 60 min
exposition. In addition, two freeze-killed tissues were treated with H2O(for 3 min and for 60 min exposition). After 3 min and 60 min
of incubation (37±1°C, 5±1% CO2, moistened), the test substance was rinsed and tissues were incubated with MTT solution in the same
manner as viable tissues in MTT test. Two hours extraction in isopropyl alcohol with shaking and OD measuring at 570 nm followed then.
Results are given in table 1.
Table 1:Direct MTT reduction in frozen tissues
Treatment |
OD570 |
% NC |
|||||
tissues |
mean |
SD |
99% confidence interval |
||||
1 |
2 |
||||||
H2O 3 min |
0.155 |
0.124 |
0.140 |
0.015 |
0.109 |
0.171 |
100.0 |
C1 3 min |
0.162 |
0.144 |
0.153 |
0.009 |
0.135 |
0.171 |
109.7 |
H2O 60 min |
0.107 |
0.080 |
0.094 |
0.014 |
0.067 |
0.121 |
100.0 |
C1 60 min |
0.084 |
0.067 |
0.076 |
0.008 |
0.059 |
0.092 |
80.7 |
Mean OD570value of treated tissues after 3 min treatment (0.153) fell into confidence interval OD570of negative control (0.109-0.171) and
average OD570value of treated tissues after 60 min treatment (0.076) fell into confidence interval OD570of negative control (0.067-0.121),
so the test substance did not reduce MTT directly, therefore it was not necessary to correct the results of MTT test.
1.3. MTT test
The procedure is described in chapter 3.6.3.
OD570measuring was performed after overnight extraction. Results are given in the following table 2.
Table 2:MTT test results (viable tissues)
time |
treatment |
OD570 |
%NC |
||||||
tissues |
mean |
SD |
CV |
||||||
1 |
2 |
3 |
|||||||
|
NC |
water |
1.270 |
1.129 |
1.217 |
1.205 |
0.058 |
0.048 |
100.0 |
3 min |
C1 |
Hysperse 12 |
1.181 |
1.314 |
1.464 |
1.320 |
0.116 |
0.088 |
109.5 |
|
PC |
8N KOH |
0.265 |
0.225 |
0.338 |
0.276 |
0.047 |
0.170 |
22.9 |
|
NC |
water |
1.416 |
1.326 |
1.379 |
1.374 |
0.037 |
0.027 |
100.0 |
60 min |
C1 |
Hysperse 12 |
1.056 |
1.195 |
0.900 |
1.050 |
0.120 |
0.115 |
76.5 |
|
PC |
8N KOH |
0.187 |
0.130 |
0.152 |
0.156 |
0.023 |
0.150 |
11.4 |
Notes to tables 1 and 2:
NC |
negative control - solvent
|
|
PC |
positive control N
|
|
C1 |
test substance
|
|
mean |
arithmetic mean |
|
% NC |
viability of single tissues compared with negative control |
|
SD |
standard deviation calculated from individual % tissue viabilities
|
|
CV |
coefficient of variance
|
Applicant's summary and conclusion
- Interpretation of results:
- GHS criteria not met
- Conclusions:
- Substance is not corrosive.
- Executive summary:
The test substance, was assayed for the in vitro skin corrosion in human epidermal model EpiDermTM. The test was performed according to Method B.40. Skin corrosion (in vitro),Council Regulation (EC) No.440/2008.
The test substance (25 mg) was placed atop the previously moistened tissue. Length of exposition was 3 and 60 minutes. Nine tissues were used for the experiment in each time, three per test substance (C1), three for positive control (PC) and three for negative control (NC). After rinsing, tissues were incubated with MTT for three hours and extracted overnight subsequently at room temperature without shaking. OD570of isopropyl alcohol extracts was measured on a spectrophotometer. Relative cell viability was calculated for each tissue as % of the mean viability of the negative control tissues.
As the test substance was suspected to be direct reducing, the test with frozen tissues was performed to detect if the test substance actually reduces MTT in tissues directly. Because this test excluded initial suspicion, the correction of MTT test results has not been made.
Under the above-described experimental design, average viability of tissues treated by the test substance, was 109.5 % of negative control average value after 3 min treatment and 76.5 % after 60 min treatment.
In the experiment arrangement given above, the test substance was non-corrosive in EpiDermTMmodel.
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.
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