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EC number: 246-332-1 | CAS number: 24593-34-8
- 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
![](https://chesar.echa.europa.eu/o/diss-blank-theme/images/factsheets/A-REACH/factsheet/print_toxicological-information.png)
Endpoint summary
Administrative data
Description of key information
skin irritation: not irritating (OECD 439; GLP)
eye irritation: not irritating (OECD 437; GLP)
Key value for chemical safety assessment
Skin irritation / corrosion
Link to relevant study records
- Endpoint:
- skin irritation: in vitro / ex vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2018-01-24 to 2018-01-26
- 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:
- 2015-07-28
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: MatTek Corporation Protocol for: In Vitro EpiDermTM Skin Irritation Test (EPI-200-SIT) For use with MatTek Corporation’s Reconstructed Human Epidermal Model EpiDerm (EPI-200-SIT)
- Version / remarks:
- 2014-11-07
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- signed 2015-06-05
- Specific details on test material used for the study:
- STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature in a sealed container - Test system:
- human skin model
- Source species:
- human
- Cell type:
- other: normal, human epidermal keratinocytes
- Cell source:
- other: humans
- Source strain:
- other: not applicable
- Details on animal used as source of test system:
- not applicable
- Justification for test system used:
- This test uses the EpiDerm™ reconstructed human epidermis model (MatTek) which consists of normal human-derived epidermal keratinocytes (NHEK) and therefore represents in vitro the target organ of the species of interest and closely resembles the biochemical and physiological properties of the upper parts of the human skin, i.e. the epidermis.
- Vehicle:
- other: Dulbecco's phosphate buffered saline
- Details on test system:
- RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDermTM (EPI-200-SIT; MatTek)
- Tissue lot number: 25875
TEST FOR DIRECT MTT REDUCTION
- to check the non-specific MTT-reducing capability of the test item 25 mg of the test item were mixed per 1 mL MTT medium and incubated for 60 min at 37 ± 1 °C in the incubator (5 % CO2, 95 % RH).
- untreated MTT medium was used as control.
- if the mixture turns blue/purple, the test item is presumed to have reduced MTT and the part of absorption due to the non-specific reduction of MTT (NSMTT) was determined by using freeze-killed tissues.
TEST FOR COLOUR INTERFERENCE
- to check the colouring potential of the test item 25 mg of the test item were mixed per 300 µL aqua dest. and per 300 µL isopropanol each in a transparent recipient and incubated at 37 ± 1°C for 60 min (5 % CO2, 95 % RH).
- the mixture showed no colouring detected by unaided eye assessment. So the additional functional test with viable tissues and the quantitative corrections were not necessary.
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 ± 1 °C for 35 ± 1 minutes followed by incubation at room temperature until the 60 ± 1 minute treatment period was completed
- Temperature of post-treatment incubation: 37 ± 1 °C
REMOVAL OF TEST MATERIAL AND CONTROLS
- after the end of the treatment period the tissues were washed 15 times with DPBS.
- subsequently, the inserts were submerged three times in DPBS and shaken to remove rests of the test item.
- then inserts were rinsed once from the inside and the outside with sterile DPBS.
- inserts were placed in prepared 6-well plates containing pre-warmed fresh assay medium per well.
- plates were post-incubated at 37 ± 1 °C, 5.0% CO2, humidified to 95%, for 24 ± 2 h. Following this incubation the tissues were transferred to new wells containing fresh assay medium and incubated for additional 18 ± 2 h.
MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL (300 µL/well)
- Incubation time: 3 hours ± 5 minutes
- Extraction of formazan: after the MTT incubation period, the tissues were rinsed three times with DPBS and allowed to dry. The tissues were transferred into 12-well plates and immersed in 2 mL isopropanol, sealed to inhibit evaporation. Extraction was carried out protected from light at room temperature at least for 2 hours with shaking on a plate shaker.
Before using the extracts, the plate had been shaken for at least 15 minutes on a plate shaker and the inserts were pierced with an injection needle. The extract was pipetted up and down 3 times before 2 x 200 µL aliquots per each tissue were transferred into a 96-well plate. Optical density (OD) was measured with a filter band without reference wavelength in a plate spectrophotometer using isopropanol as a blank.
- Wavelength: 570 nm
- Filter bandwidth: maximum ± 30 nm
FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: tissues pass analysis for tissue viability
- Barrier function: tissues pass analysis for tissue functionality
- Morphology: presence of a functional stratum corneum, a viable basal cell layer, and intermediate spinous and granular layers.
- Contamination: absence of bacteria, yeast, and other fungi (long term antibiotic, antimycotic free culture) as well as absence of HIV1-virus, Hepatitis B virus and Hepatitis C virus
Please also refer to the field "Attached background material" below.
PREDICTION MODEL / DECISION CRITERIA
The mean optical density (OD) of the three negative control tissues was calculated after blank correction. This value corresponded to 100 % tissue viability in the current test. For each individual tissue treated with the test item or the positive control, the individual relative tissue viability is calculated according to the following formula:
Relative viability (%) = [(mean ODtest item / positive control) / ODmean of negative control] * 100
For the test item and the positive control the mean relative viability ± relative standard deviation of the three individual tissues were calculated and used for classification according to the following prediction model:
Irritant potential of the test item was predicted from the relative mean tissue viabilities compared to the negative control tissues concurrently treated with DPBS. The test item is considered to be irritant to skin in accordance with regulation EC 1272/2008 (UN GHS “Category 2”), if the tissue viability after exposure and post-incubation is less or equal to 50%. Further testing is required to resolve between UN GHS categories 1 and 2 and decide on the final classification of the test substance. The test substance may be considered as non-irritant to skin in accordance with regulation EC 1272/2008 and UN GHS “No Category” if the tissue viability after exposure and post-treatment incubation is more than 50%. - 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 (39 mg/cm²) of the test item
Firstly, 25 µL of the vehicle were applied to the epidermal surface in order to improve the contact between the powder and the epidermis. Afterwards, 25 mg (39 mg/cm²) of the test item were applied directly atop the EpiDermTM tissue using an application spoon avoiding compression of the test item. The test item was spread to match size of the tissue by using a bulb-headed Pasteur pipette.
VEHICLE
- Amount(s) applied (volume or weight with unit): 25 µL of the vehicle
NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 30 µL DPBS
POSITIVE CONTROL
- Amount(s) applied (volume or weight): 30 µL of 5 % SDS solution - Duration of treatment / exposure:
- 60 ± 1 minute
- Duration of post-treatment incubation (if applicable):
- approx. 42 hours
- Number of replicates:
- triplicates
- Irritation / corrosion parameter:
- % tissue viability
- Remarks:
- (mean)
- Value:
- 104.3
- Vehicle controls validity:
- not examined
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Other effects / acceptance of results:
- TEST FOR MTT INTERFERENCE
The mixture of 25 mg test item per 1 mL MTT medium showed no reduction of MTT compared to the solvent. The mixture did not turn blue/purple. Therefore, NSMTT equalled 0%.
TEST FOR COLOUR INTERFERENCE
The mixture of 25 mg of the test item per 300 μl aqua dest. and/or per 300 μL isopropanol showed no colouring detectable by unaided eye-assessment. Therefore, NSC equalled 0%.
ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: mean absolute OD570 of the three negative control tissues was ≥ 0.8 and ≤ 2.8 (value: 1.845).
- Acceptance criteria met for positive control: mean relative tissue viability (% negative control) of the positive control was ≤ 20% (4.1 %)
- Acceptance criteria met for variability between replicate measurements: standard deviation of viability of replicate tissues of all dose groups was ≤ 18% (0.6 % - 6.2 %).
- The absorbance values were not below historically established boundaries.
Please also refer to the field "An other information on results incl. tables" below. - Interpretation of results:
- GHS criteria not met
- Conclusions:
- In this study under the reported conditions of the human skin model test, 2-ethylhexanoic acid, cerium salt is not irritant to the skin (No Category) according to Regulation (EC) No. 1272/2008 and UN GHS.
Reference
Table1: Result of the Test Item 2-ethylhexanoic acid, cerium salt
Name |
Negative Control |
Positive Control |
Test Item |
||||||
Tissue |
1 |
2 |
3 |
1 |
2 |
3 |
1 |
2 |
3 |
Absolute OD570 |
1.717 |
1.912 |
1.872 |
0.106 |
0.124 |
0.125 |
1.906 |
1.837 |
2.054 |
1.743 |
1.895 |
1.929 |
0.107 |
0.125 |
0.124 |
1.878 |
1.820 |
2.036 |
|
Mean Absolute OD570 |
1.845**** |
0.119 |
1.922 |
||||||
OD570(Blank Corrected) |
1.673 |
1.868 |
1.828 |
0.062 |
0.080 |
0.081 |
1.862 |
1.793 |
2.010 |
1.699 |
1.851 |
1.885 |
0.063 |
0.081 |
0.081 |
1.834 |
1.776 |
1.992 |
|
Mean OD570of the Duplicates (Blank Corrected) |
1.686 |
1.860 |
1.857 |
0.063 |
0.081 |
0.081 |
1.848 |
1.785 |
2.001 |
Total Mean OD570of 3 Replicate Tissues (Blank Corrected) |
1.801* |
0.075 |
1.878 |
||||||
SD of Mean OD570of the Replicates (Blank Corrected) |
0.099 |
0.010 |
0.111 |
||||||
Relative Tissue Viability [%] |
93.6 |
103.3 |
103.1 |
3.5 |
4.5 |
4.5 |
102.6 |
99.1 |
111.1 |
Mean Relative Tissue Viability [%] |
100.0 |
4.1** |
104.3 |
||||||
SD of Relative Tissue Viability [%] |
5.5*** |
0.6*** |
6.2*** |
||||||
CV [% Viabilities] |
5.5 |
13.9 |
5.9 |
* Blank-corrected mean OD570 nmof the negative control corresponds to 100% absolute tissue viability.
** Mean relative tissue viability of the three positive control tissues is ≤ 20%.
*** Standard deviation (SD)of relative tissue viabilityobtained from the three concurrently tested tissuesfor test item, positive and negative controlis ≤ 18%
****Mean absolute OD570 of the negative control tissues is ≥0.8 and ≤2.8.
Table2: Historical Data
|
Mean Absolute OD570±30nmNC |
MeanAbsoluteOD570±30nmPC |
Mean Relative Viability [%] PC |
SD Viability [%] |
Mean |
1.861 |
0.114 |
3.7 |
4.4 |
SD |
0.247 |
0.033 |
1.5 |
4.1 |
Range of |
1.367 – 2.355 |
0.048 – 0.181 |
0.7 – 6.8 |
0.0 – 12.5 |
n |
25 |
25 |
25 |
117 |
LCL: Lower control limit (95%, mean – 2*SD)
UCL: Upper control limit (95%, mean + 2*SD)
n: number of control values
Historical data were generated in 2017.
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (not irritating)
Eye irritation
Link to relevant study records
- Endpoint:
- eye irritation: in vitro / ex vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2018-02-14
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)
- Version / remarks:
- 2017-10-09
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- signed 2015-06-05
- Specific details on test material used for the study:
- STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature in a sealed container
TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: test item was crushed to a fine powder using a mortar and a pestle. - Details on test animals or tissues and environmental conditions:
- SOURCE OF COLLECTED EYES
- Source: abattoir A. Moksel AG, Buchloe, Germany
- Storage, temperature and transport conditions of ocular tissue: fresh eyes were collected from the slaughterhouse and were transported in HBSS containing Pen/Strep on ice to the laboratories.
- Time interval prior to initiating testing: immediately after arrival of the eyes, cornea preparation was initiated and was used for BCOP testing on the same day. - Vehicle:
- physiological saline
- Controls:
- yes, concurrent positive control
- yes, concurrent negative control
- Amount / concentration applied:
- TEST MATERIAL
- Amount(s) applied (volume or weight with unit): about 750 mg of the test item
Since it was not possible to get a solution with the test item, it was administered directly and moistened with a drop of physiological saline (0.9% NaCl). - Duration of treatment / exposure:
- 4 hours
- Observation period (in vivo):
- not applicable
- Duration of post- treatment incubation (in vitro):
- not required
- Number of animals or in vitro replicates:
- Number of bovine corneae per dose:
Test item: triplicates
Negative control: triplicates
Positive control: triplicates - Details on study design:
- SELECTION AND PREPARATION OF CORNEAS
- eyes were examined for defects and any defective eyes were discarded. Eyes with scratches or any kind of opacity were not used.
- tissue surrounding the eyeball was pulled away and the cornea was excised.
- isolated corneas were stored in a petri dish containing HBSS. Before the corneas were mounted in corneal holders (Duratec GmbH) with the endothelial side against the O-ring of the posterior chamber, they had been visually examined for defects and any defective cornea had been discarded. The anterior chamber was then positioned on top of the cornea and tightened with screws. The chambers of the corneal holder were then filled with pre-warmed MEMI (without phenol red) containing 1% FBS and 2 mM L-glutamine (complete MEMI).
- corneas were incubated for one hour at 32 ± 1 °C for equilibration in an air incubator.
QUALITY CHECK OF THE ISOLATED CORNEAS
- after the equilibration period, the medium was removed from both chambers and replaced with fresh complete MEM.
- an initial measurement was performed on each of the corneas using the opacitometer.
- three corneas with illuminance readings approximately equivalent to the median illuminance of all corneas were selected as negative-control corneas.
- the illuminance of each cornea was read and recorded.
- only corneas that had an initial illuminance reading I > I0/1.1651 lux (an equivalent to the opacity threshold of 7 as listed in OECD 437) were used for the assay.
APPLICATION DOSE AND EXPOSURE TIME
- medium was removed from the anterior chamber and replaced with the test item or control.
- 750 µL of the control substances (closed-chamber method) and enough test item to completely cover the cornea (about 750 mg, open-chamber method) were introduced into the anterior chamber.
- after 4 hours ± 5 minutes incubation at 32 ± 1 °C either the test substance or the control substance was removed.
REMOVAL OF TEST SUBSTANCE/CONTROL SUBSTANCES
- epithelium was washed more than three times with MEM (containing phenol red), since the corneas showed small test item residues, which could not be removed.
- the cornea was finally rinsed with complete MEM (without phenol red).
METHODS FOR MEASURED ENDPOINTS:
- anterior chamber was refilled with complete MEM and an illuminance measurement was performed.
- corneas were visually examined for tissue peeling, residual test chemical and non-uniform opacity patterns and observation were recorded.
- after the illuminance measurement was performed, the medium was removed from both chambers of the holder.
- posterior chamber was refilled with fresh complete MEMI.
- 1 mL of a 5 mg/mL sodium fluorescein solution was added to the anterior chamber and the corneas were incubated for 90 minutes at 32 ± 1 °C in horizontal position.
- then the medium from the posterior chamber was removed and its optical density at 490 nm (OD490) was determined, using a spectrophotometer (Jenway 6405 UV/VIS).
Evaluation of the opacity:
- the following formula was used to calculate the opacity, whereas the values a and b are equipment-specific variables empirically determined by the manufacturer:
Opacity = ((I0/I) - b)/ a
with a = 0.025 and b = 0.9894
- value I0 is the illuminance through a holder without cornea, but with windows and liquid. This value is determined by taking the mean for a set of cornea holders and is reevaluated periodically. This I0 value was than calculated to the respective data of the opacitometer and the data according to guideline (opacity < 7). So the initial illuminance could be calculated and corneas below this value were discarded.
- change in opacity for each cornea (test item, positive and negative control) was calculated by subtracting the initial opacity reading from the final opacity reading. These values of test item treated cornea or positive control were corrected by subtracting from each the average change in opacity observed for the negative-control corneas to obtain a corrected opacity. The mean corrected opacity value for each treatment was calculated by averaging the corrected opacity values of each cornea for a given treatment.
Evaluation of the permeability:
- mean OD490 for the blank cuvettes was calculated.
- mean blank OD490 was subtracted from the OD490 of each cuvette (corrected OD490).
- any dilutions that were made to bring the OD490 values into the linear range of the spectrophotometer (OD490 should be less than 1.500), were taken into account by multiplying the OD490 value of the dilution by the dilution factor.
- final-corrected OD490 of the test article and the positive control were calculated by subtracting the average-corrected OD490 of the negative-control corneas from the corrected OD490 value of each treated cornea:
Final-corrected OD490 = (OD490 – mean blank OD490) – average-corrected negative control OD490
- mean OD490 value of each treatment group was calculated by averaging the final corrected OD490 values of the treated corneas for that treatment condition.
SCORING SYSTEM: In Vitro Irritancy Score (IVIS)
The following formula was used to determine the in vitro irritation score (IVIS):
IVIS = mean opacity value + (15 x mean permeability OD490 value)
To determine the IVIS of the positive control and the test item, the corrected opacity and OD490 values are used.
ACCEPTABILITY CRITERIA
- the BCOP assay is considered to be valid if the in vitro irritation score obtained with the positive control falls within the two standard deviations of the current historical mean.
- the negative control responses should result in opacity and permeability values that are less than the established upper limits for background bovine corneas treated with the respective negative control. - Irritation parameter:
- in vitro irritation score
- Remarks:
- (mean)
- Run / experiment:
- Experiment 1
- Value:
- 2.13
- Vehicle controls validity:
- not examined
- Negative controls validity:
- valid
- Positive controls validity:
- not valid
- Remarks on result:
- other: The in vitro irritation score obtained with the positive control did not fall within the two standard deviations of the current historical mean. Therefore, the assy was not considered to be valid and the experiment was repeated (Experiment 2).
- Irritation parameter:
- in vitro irritation score
- Remarks:
- (mean)
- Run / experiment:
- Experiment 2
- Value:
- 2.58
- Vehicle controls validity:
- not examined
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Other effects / acceptance of results:
- EXPERIMENT 1:
- Measurement after treatment:
All 3 corneas treated with 2-ethylhexanoic acid, cerium salt showed very slight opacity of the tissue.
- Acceptance of results:
- Acceptance criteria met for negative control: the negative control responses resulted in opacity and permeability values that are less than the established upper limits for background bovine corneas treated with the respective negative control.
- Acceptance criteria met for positive control: the in vitro irritation score obtained with the positive control did not fall within the two standard deviations of the current historical mean. Therefore, the assy was not considered to be valid and the experiment was repeated.
EXPERIMENT 2:
- Visual Observation after treatment:
None of the 3 corneas treated with 2-ethylhexanoic acid, cerium salt showed any opacity of the tissue. 2 corneas showed small test item residues that could not be removed by non-invasive methods, such as additional rinsing. 2 test item treated corneas showed small test item residues, that could not be removed by non-invasive methods, such as additional rinsing. An interference of residual test material with the opacity measurement is not assumed, since the residues were very small and the qualitative visual appraisal of the test item treated corneas showed no opacity of the tissue, so that a possible impact on the opacity measurement can be disregarded. Furthermore, relative to the negative control, the test item caused no increase of corneal opacity and permeability in all 3 corneas. In conclusion, the residual material has no effect on the result.
- Measurement after treatment:
Relative to the negative control, the test item caused no increase of corneal opacity and permeability in all 3 corneas.
- Acceptance of results:
The test is valid since the IVIS of the positive control falls within two standard deviations of the current historical mean and opacity and permeability of the negative control are less than the respective established upper limits for background opacity and permeability.
Please also refer for results to the field "Any other information on results incl. tables" below - Interpretation of results:
- GHS criteria not met
- Conclusions:
- The test substance is not classified as serious eye damaging or eye irritant according to the Regulation (EC) No 1272/2008 and its subsequent amendments and corrections.
Reference
EXPERIMENT 2 valid:
Table 1: Opacity
Cornea |
Test Item |
Initial |
Final |
Change of |
Corrected |
|
1 |
Negative |
0.19 |
0.97 |
0.79 |
|
|
2 |
0.19 |
0.97 |
0.79 |
|
||
3 |
0.12 |
0.66 |
0.54 |
|
||
MV |
0.16 |
0.87 |
0.71 |
|
||
4 |
Positive |
1.40 |
99.43 |
98.03 |
97.33 |
|
5 |
1.43 |
79.29 |
77.86 |
77.15 |
||
6 |
1.01 |
105.47 |
104.46 |
103.75 |
||
MV |
1.28 |
94.73 |
93.45 |
92.74 |
||
7 |
Test Item |
-0.18 |
1.04 |
1.23 |
0.52 |
|
8 |
-1.24 |
5.61 |
6.85 |
6.15 |
||
9 |
0.73 |
2.09 |
1.36 |
0.65 |
||
MV |
-0.23 |
2.92 |
3.15 |
2.44 |
||
MV = mean value |
Table 2: Permeability
Cornea |
Test Item |
OD490 |
Corrected |
|
1 |
Negative |
0.006 |
|
|
2 |
0.005 |
|
||
3 |
0.005 |
|
||
MV |
0.005 |
|
||
4 |
Positive |
2.715 |
2.710 |
|
5 |
3.240 |
3.235 |
||
6 |
5.280 |
5.275 |
||
MV |
3.745 |
3.740 |
||
7 |
Test Item |
0.013 |
0.008 |
|
8 |
0.009 |
0.004 |
||
9 |
0.022 |
0.017 |
||
MV |
0.015 |
0.009 |
||
MV = mean value |
Table 3: In Vitro Irritation Score
Cornea |
Test Item |
Corrected |
Corrected |
IVIS |
|
1 |
Negative |
0.79 |
0.006 |
|
|
2 |
0.79 |
0.005 |
|
||
3 |
0.54 |
0.005 |
|
||
MV |
0.71 |
0.005 |
0.79 |
||
4 |
Positive |
97.33 |
2.710 |
|
|
5 |
77.15 |
3.235 |
|
||
6 |
103.75 |
5.275 |
|
||
MV |
92.74 |
3.740 |
148.84 |
||
7 |
Test Item |
0.52 |
0.008 |
|
|
8 |
6.15 |
0.004 |
|
||
9 |
0.65 |
0.017 |
|
||
MV |
2.44 |
0.009 |
2.58 |
||
MV = mean value |
Table 4: Historical mean In Vitro Irritation Score of the Positive Control (Imidazole 20%) from February 2015 until May 2018
|
IVIS Positive Control - Imidazole 20 % |
|
Mean Value (MV) |
123.98 |
|
Standard Deviation (SD) |
17.69 |
|
MV- 2xSD |
88.60 |
|
MV+2xSD |
159.37 |
|
Number of Replicates providing Historical Mean: 36 |
||
Positive controls are updated after every single experiment or at least every 3 months.
Table 5: Historical Data on Opacity and Permeability of the Positive Control (Imidazole 20%) from August 2017 until May 2018
Replicates Providing Historical Mean |
Cornea No. |
Opacity |
Permeability |
IVIS |
||
Change of |
Corrected |
OD490 Value |
Corrected |
|||
1 |
4 |
122.785 |
121.861 |
0.662 |
0.624 |
133.420 |
5 |
117.173 |
116.249 |
1.220 |
1.182 |
||
6 |
102.655 |
101.731 |
2.260 |
2.222 |
||
2 |
4 |
108.553 |
106.381 |
1.473 |
1.450 |
123.050 |
5 |
79.491 |
77.319 |
2.465 |
2.442 |
||
6 |
83.618 |
81.446 |
3.065 |
3.042 |
||
3 |
4 |
55.644 |
56.308 |
2.200 |
2.189 |
92.540 |
5 |
71.511 |
72.175 |
1.348 |
1.337 |
||
6 |
65.148 |
65.812 |
2.040 |
2.029 |
||
4 |
4 |
68.39 |
67.72 |
2.400 |
2.383 |
112.690 |
5 |
70.88 |
70.21 |
2.810 |
2.793 |
||
6 |
94.23 |
93.56 |
1.945 |
1.928 |
||
5 |
4 |
70.53 |
70.35 |
1.296 |
1.284 |
102.440 |
5 |
78.68 |
78.50 |
1.363 |
1.351 |
||
6 |
91.69 |
91.51 |
1.840 |
1.828 |
||
6 |
4 |
95.54 |
94.92 |
1.478 |
1.467 |
114.650 |
5 |
83.58 |
82.96 |
1.500 |
1.489 |
||
6 |
91.11 |
90.49 |
2.095 |
2.084 |
||
7 |
4 |
89.35 |
88.86 |
2.855 |
2.838 |
149.420 |
5 |
117.36 |
116.87 |
2.215 |
2.198 |
||
6 |
130.15 |
129.66 |
2.505 |
2.488 |
||
8 |
4 |
80.99 |
80.24 |
2.140 |
2.127 |
120.110 |
5 |
78.40 |
77.65 |
3.070 |
3.057 |
||
6 |
76.27 |
75.51 |
3.290 |
3.277 |
||
9 |
4 |
98.03 |
97.33 |
2.715 |
2.710 |
148.838 |
5 |
77.86 |
77.15 |
3.240 |
3.235 |
||
6 |
104.46 |
103.75 |
5.280 |
5.275 |
||
Mean Value (MV) |
89.040 |
88.390 |
2.251 |
2.234 |
121.906 |
|
Standard Deviation (SD) |
18.814 |
18.573 |
0.916 |
0.919 |
19.360 |
|
MV- 2xSD |
51.412 |
51.244 |
0.419 |
0.396 |
83.187 |
|
MV+2xSD |
126.667 |
125.536 |
4.082 |
4.073 |
160.626 |
Table 6: Historical meanIn Vitro Irritation Score of the Negative Control (NaCl 0.9 %) from February 2015 until May 2018
|
IVIS Negative Control - NaCl 0.9 % |
|
Mean Value (MV) |
1.09 |
|
Standard Deviation (SD) |
0.76 |
|
MV- 2xSD |
-0.44 |
|
MV+2xSD |
2.62 |
|
Number of Replicates providing Historical Mean: 36 |
Negative controls are updated after every single experiment or at least every 3 months.
Table 7: Historical Data on Opacity and Permeability of the Negative Control (NaCl 0.9 %) from August 2017 until May 2018
Replicates Providing Historical Mean |
Cornae No. |
Opacity |
Permeability |
IVIS |
Change of |
OD490 Value |
|||
1 |
1 |
0.234 |
0.008 |
1.49 |
2 |
1.738 |
0.008 |
||
3 |
0.800 |
0.098 |
||
2 |
1 |
0.978 |
0.019 |
2.52 |
2 |
3.920 |
0.022 |
||
3 |
1.617 |
0.028 |
||
3 |
1 |
-0.149 |
0.009 |
-0.50 |
2 |
-0.415 |
0.015 |
||
3 |
-1.427 |
0.009 |
||
4 |
1 |
0.776 |
0.008 |
0.92 |
2 |
0.808 |
0.022 |
||
3 |
0.418 |
0.020 |
||
5 |
1 |
0.035 |
0.010 |
0.35 |
2 |
0.036 |
0.013 |
||
3 |
0.466 |
0.012 |
||
6 |
1 |
1.030 |
0.017 |
0.79 |
2 |
0.190 |
0.008 |
||
3 |
0.640 |
0.009 |
||
7 |
1 |
0.714 |
0.024 |
0.74 |
2 |
0.373 |
0.015 |
||
3 |
0.371 |
0.012 |
||
8 |
1 |
1.034 |
0.013 |
0.94 |
2 |
0.596 |
0.01 |
||
3 |
0.623 |
0.015 |
||
9 |
1 |
0.789 |
0.006 |
0.79 |
2 |
0.789 |
0.005 |
||
3 |
0.543 |
0.005 |
||
Mean Value (MV) |
0.649 |
0.016 |
0.893 |
|
Standard Deviation (SD) |
0.894 |
0.017 |
0.813 |
|
MV- 2xSD |
-1.140 |
-0.019 |
-0.732 |
|
MV+2xSD |
2.438 |
0.051 |
2.519 |
EXPERIMENT 1 invalid:
Table 1: Opacity
Cornea |
Test Item |
Initial |
Final |
Change of |
Corrected |
|
1 |
Negative |
2.92 |
3.03 |
0.12 |
|
|
2 |
2.61 |
2.99 |
0.38 |
|
||
3 |
2.61 |
2.73 |
0.11 |
|
||
MV |
2.71 |
2.92 |
0.20 |
|
||
4 |
Positive |
3.30 |
221.22 |
217.91 |
217.71 |
|
5 |
3.62 |
72.19 |
68.57 |
68.37 |
||
6 |
3.58 |
81.13 |
77.55 |
77.35 |
||
MV |
3.50 |
124.85 |
121.35 |
121.14 |
||
7 |
Test Item |
2.27 |
4.89 |
2.61 |
2.41 |
|
8 |
1.36 |
4.72 |
3.36 |
3.15 |
||
9 |
3.07 |
3.78 |
0.71 |
0.50 |
||
MV |
2.24 |
4.46 |
2.23 |
2.02 |
||
MV = mean value
Table 2: Permeability
Cornea |
Test Item |
OD490 |
Corrected |
|
1 |
Negative |
0.015 |
|
|
2 |
0.018 |
|
||
3 |
0.026 |
|
||
MV |
0.020 |
|
||
4 |
Positive |
2.625 |
2.605 |
|
5 |
3.210 |
3.190 |
||
6 |
2.945 |
2.925 |
||
MV |
2.927 |
2.907 |
||
7 |
Test Item |
0.026 |
0.006 |
|
8 |
0.034 |
0.014 |
||
9 |
0.021 |
0.001 |
||
MV |
0.027 |
0.007 |
||
MV = mean value |
Table 3: In vitro irritation score
Cornea |
Test Item |
Corrected |
Corrected |
IVIS |
|
1 |
Negative |
0.12 |
0.015 |
|
|
2 |
0.38 |
0.018 |
|
||
3 |
0.11 |
0.026 |
|
||
MV |
0.20 |
0.020 |
0.50 |
||
4 |
Positive |
217.71 |
2.605 |
|
|
5 |
68.37 |
3.190 |
|
||
6 |
77.35 |
2.925 |
|
||
MV |
121.14 |
2.907 |
164.75 |
||
7 |
Test Item |
2.41 |
0.006 |
|
|
8 |
3.15 |
0.014 |
|
||
9 |
0.50 |
0.001 |
|
||
MV |
2.02 |
0.007 |
2.13 |
||
MV = mean value |
Table 4: Historical mean in vitro irritation score of the positive control from February 2015 until January 2018
|
IVIS Positive Control - Imidazole 20 % |
|
Mean Value (MV) |
122.57 |
|
Standard Deviation (SD) |
17.33 |
|
MV- 2xSD |
87.91 |
|
MV+2xSD |
157.23 |
|
Number of Replicates providing Historical Mean: 33 |
||
Positive controls are updated after every single experiment or at least every 3 months.
Table 5: Historical data on opacity and permeability of the positive control (Imidazole 20 %) from August 2017 until January 2018
|
|
|
Incubation: 240 min |
|
|
|
|
|
Number of Replicates |
Cornea No. |
Opacity |
Permeability |
IVIS |
||
|
Change of Value |
Corrected Value |
OD490 Value |
Corrected Value |
|||
2017 |
1 |
4 |
122.785 |
121.861 |
0.662 |
0.624 |
133.420 |
5 |
117.173 |
116.249 |
1.220 |
1.182 |
|||
6 |
102.655 |
101.731 |
2.260 |
2.222 |
|||
2 |
4 |
108.553 |
106.381 |
1.473 |
1.450 |
123.050 |
|
5 |
79.491 |
77.319 |
2.465 |
2.442 |
|||
6 |
83.618 |
81.446 |
3.065 |
3.042 |
|||
3 |
4 |
55.644 |
56.308 |
2.200 |
2.189 |
92.540 |
|
5 |
71.511 |
72.175 |
1.348 |
1.337 |
|||
6 |
65.148 |
65.812 |
2.040 |
2.029 |
|||
4 |
4 |
68.39 |
67.72 |
2.400 |
2.383 |
112.690 |
|
5 |
70.88 |
70.21 |
2.810 |
2.793 |
|||
6 |
94.23 |
93.56 |
1.945 |
1.928 |
|||
5 |
4 |
70.53 |
70.35 |
1.296 |
1.284 |
102.440 |
|
5 |
78.68 |
78.50 |
1.363 |
1.351 |
|||
6 |
91.69 |
91.51 |
1.840 |
1.828 |
|||
2018 |
6 |
4 |
95.54 |
94.92 |
1.478 |
1.467 |
114.650 |
5 |
83.58 |
82.96 |
1.500 |
1.489 |
|||
6 |
91.11 |
90.49 |
2.095 |
2.084 |
|||
|
Mean Value (MV) |
86.178 |
85.528 |
1.859 |
1.840 |
113.132 |
|
|
Standard Deviation (SD) |
18.438 |
18.015 |
0.617 |
0.619 |
14.497 |
|
|
MV- 2xSD |
49.303 |
49.499 |
0.624 |
0.603 |
84.138 |
|
|
MV+2xSD |
123.053 |
121.558 |
3.094 |
3.078 |
142.126 |
Table 6: Historical mean in vitro irritation score of the negative control from February 2015 until January 2018
IVIS Negative Control – NaCl 0.9 % |
|
Mean Value (MV) |
1.11 |
Standard Deviation (SD) |
0.79 |
MV-2xSD |
-0.47 |
MV+2xSD |
2.70 |
Number of Replicates providing Historical Mean: 33 |
Negative controls are updated after every single experiment or at least every 3 months
Table 7: Historical data on opacity and permeability of the negative control (NaCl 0.9 %) from August 2017 until January 2018
Incubation: 240 min |
|||||
Number of Replicates Providing Historical Mean |
Cornae No. |
Opacity |
Permeability |
IVIS |
|
Change of |
OD490 Value |
||||
2017 |
1 |
1 |
0.234 |
0.008 |
1.49 |
2 |
1.738 |
0.008 |
|||
3 |
0.800 |
0.098 |
|||
2 |
1 |
0.978 |
0.019 |
2.52 |
|
2 |
3.920 |
0.022 |
|||
3 |
1.617 |
0.028 |
|||
3 |
1 |
-0.149 |
0.009 |
-0.50 |
|
2 |
-0.415 |
0.015 |
|||
3 |
-1.427 |
0.009 |
|||
4 |
1 |
0.776 |
0.008 |
0.92 |
|
2 |
0.808 |
0.022 |
|||
3 |
0.418 |
0.020 |
|||
5 |
1 |
0.035 |
0.010 |
0.35 |
|
2 |
0.036 |
0.013 |
|||
3 |
0.466 |
0.012 |
|||
2018 |
6 |
1 |
1.030 |
0.017 |
0.79 |
2 |
0.190 |
0.008 |
|||
3 |
0.640 |
0.009 |
|||
Mean Value (MV) |
0.650 |
0.019 |
0.928 |
||
Standard Deviation (SD) |
1.097 |
0.021 |
1.024 |
||
MV- 2xSD |
-1.543 |
-0.023 |
-1.120 |
||
MV+2xSD |
2.843 |
0.060 |
2.976 |
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (not irritating)
Respiratory irritation
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
Skin irritation:
The substance was not observed to be a skin irritant in a reliable in vitro skin irritation study according to OECD 439.
Eye irritation:
The substance was not observed to be an eye irritant in a reliable in vitro eye irritation study according to OECD 437.
Justification for classification or non-classification
Skin irritation
The substance does not possess a skin irritation potential based on an in vitro OECD 439 test and does not require classification as skin irritant according to Regulation (EC) No 1272/2008 and its subsequent adaptations.
Eye irritation
The substance does not possess an eye irritation potential based on an in vitro OECD 437 test and does not require classification as eye irritant according to Regulation (EC) No 1272/2008 and its subsequent adaptations.
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.
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