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EC number: 921-114-5 | CAS number: -
- 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
DPRA: negative
KeratinoSens: positive
hClat: positive
Key value for chemical safety assessment
Skin sensitisation
Link to relevant study records
- Endpoint:
- skin sensitisation: in chemico
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- 2017-08-11 to 2017-09-09
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 442C (In Chemico Skin Sensitisation: Direct Peptide Reactivity Assay (DPRA))
- Qualifier:
- according to guideline
- Guideline:
- other: Direct Peptide Reactivity Assay (DPRA) for Skin Sensitization Testing, DB-ALM Protocol n°154, January 12, 2013
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany
- Type of study:
- direct peptide reactivity assay (DPRA)
- Details on the study design:
- The in chemico direct peptide reactivity assay (DPRA) enables detection of the sensitising potential of a test item by quantifying the reactivity of test chemicals towards synthetic peptides containing either lysine or cysteine.
- Positive control results:
- The 100 mM stock solution of the positive control (cinnamic aldehyde) showed high reactivity towards the synthetic peptides. The mean depletion of both peptides was 65.19%.
- Key result
- Run / experiment:
- other: cysteine run
- Parameter:
- other: mean peptide depletion [%]
- Value:
- 0.06
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Run / experiment:
- other: lysine run
- Parameter:
- other: mean peptide depletion [%]
- Value:
- 0.26
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Other effects / acceptance of results:
- Acceptance Criteria
The run meets the acceptance criteria if:
- the standard calibration curve has a r² > 0.99,
- the mean percent peptide depletion (PPD) value of the three replicates for the positive control is
between 60.8% and 100% for the cysteine peptide and the maximum standard deviation (SD) for the
positive control replicates is < 14.9%,
- the mean percent peptide depletion (PPD) value of the three replicates for the positive control is
between 40.2% and 69.0% for the lysine peptide and the maximum SD for the positive control
replicates is < 11.6%,
- the mean peptide concentration of the three reference controls A replicates is 0.50 ± 0.05 mM,
- the coefficient of variation (CV) of peptide peak areas for the six reference control B replicates and three reference control C replicates in acetonitrile is < 15.0%.
The results of the test item meet the acceptance criteria if:
- the maximum standard deviation (SD) for the test chemical replicates is < 14.9% for the cysteine
percent depletion (PPD),
- the maximum standard deviation (SD) for the test chemical replicates is < 11.6% for the lysine
percent depletion (PPD),
- the mean peptide concentration of the three reference controls C replicates in the appropriate solvent is 0.50 ± 0.05 mM.
Both peptide runs and the test item results met the acceptance criteria of the test. - Interpretation of results:
- other: The result should be considered in the context of integrated approach such as IATA.
- Conclusions:
- In this study under the given conditions the test item showed minimal reactivity towards the cysteine peptide. The test item might be considered as “non-sensitiser”.
The data generated with this method may be not sufficient to conclude on the absence of skin sensitisation potential of chemicals and should be considered in the context of integrated approach such as IATA. - Executive summary:
In the present study the test material was dissolved in acetonitrile based on the results of the pre-experiments. Based on a molecular weight of 206.29 g/mol a 100 mM stock solution was prepared. The test item solutions were tested by incubating the samples with the peptides containing either cysteine or lysine for 24 ± 2 h at 25 ± 2.5 °C. Subsequently samples were analysed by HPLC.
For the 100 mM stock solution of the test item no turbidity or precipitation was observed when diluted with the cysteine peptide solution. After the 24 h±2 h incubation period but prior to the HPLC analysis samples were inspected for precipitation, turbidity or phase separation. Precipitation was observed for the samples of the positive control (excluding the co-elution control of the positive control). Samples were not centrifuged prior to the HPLC analysis.
For the 100 mM stock solution of the test item a turbidity was observed when diluted with the lysine peptide solution. After the 24 h ± 2 h incubation period but prior to the HPLC analysis samples were inspected for precipitation, turbidity or phase separation. Phase separation (small droplets) was observed for the samples of the positive control (including co-elution of the positive control) and for the samples of the test item (including co-elution control of the test item). Samples were not centrifuged prior to the HPLC analysis.
Since the acceptance criteria for the depletion range of the positive control were fulfilled, the observed precipitation and phase separation were regarded as insignificant.
No co-elution of test item with the peptide peaks was observed. However, due to the phase separation observed in the lysine peptide samples and possible underestimation of the reactivity, the sensitising potential of the test item was predicted only from the peptide depletion of the cysteine peptide by comparing the peptide concentration of the test item treated samples to the corresponding reference control C (RC Cacetonitrile).
The 100 mM stock solution of the test item showed minimal reactivity towards the synthetic peptide. The mean depletion of the cysteine peptide was ≤ 6.38% (0.06%). Based on the prediction model 2 the test item can be considered as non-sensitiser.
The 100 mM stock solution of the positive control (cinnamic aldehyde) showed high reactivity towards the synthetic peptides. The mean depletion of both peptides was 65.19%.
- Endpoint:
- skin sensitisation: in vitro
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- 2017-09-20 to 2017-10-19
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 442D (In Vitro Skin Sensitisation: ARE-Nrf2 Luciferase Test Method)
- Qualifier:
- according to guideline
- Guideline:
- other: KeratinoSens™, EURL ECVAM DB-ALM Protocol No. 155, July 1st, 2015
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany
- Type of study:
- activation of keratinocytes
- Details on the study design:
- The in vitro KeratinoSens™ assay enables detection of the sensitising potential of a test item by addressing the second molecular key event of the adverse outcome pathway (AOP), namely activation of keratinocytes, by quantifying the luciferase activity in the transgenic cell line KeratinoSens™. The luciferase activity, assessed by luminescence measurement, compared to the respective solvent controls is used to support discrimination between skin sensitisers and non-sensitisers.
- Positive control results:
- The luciferase activity induced by the positive control at a concentration of 64 µM was between 2 and 8 (4.55 in experiment 1; 6.13 in experiment 2)
- Run / experiment:
- other: 1
- Parameter:
- other: luciferase activity [fold induction]
- Value:
- 6.23
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Run / experiment:
- other: 1
- Parameter:
- other: cell viability [%]
- Value:
- 95.3
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Run / experiment:
- other: 1
- Parameter:
- other: EC1.5 [µM]
- Value:
- 6.23
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- positive indication of skin sensitisation
- Run / experiment:
- other: 2
- Parameter:
- other: luciferase activity [fold induction]
- Value:
- 4.35
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Run / experiment:
- other: 2
- Parameter:
- other: cell viability [%]
- Value:
- 91
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Run / experiment:
- other: 2
- Parameter:
- other: EC1.5 [µM]
- Value:
- 9.72
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- positive indication of skin sensitisation
- Other effects / acceptance of results:
- Acceptance Criteria
The test meets acceptance criteria if:
- the luciferase activity induction of the positive control is statistically significant above the threshold of 1.5 (using a t-test) in at least one of the tested concentrations
- the average induction in the three technical replicates for the positive control at a concentration of 64 µM is between 2 and 8
- the EC1.5 value of the positive control is within two standard deviations of the historical mean
- the average coefficient of variation (CV; consisting of 6 wells) of the luminescence reading for the negative (solvent) control DMSO is <20% in each repetition.
The controls fullfilled the validity criteria of the test. - Interpretation of results:
- other: The result should be considered in the context of integrated approach such as IATA.
- Conclusions:
- In this study under the given conditions the test item did induce the luciferase activity in the transgenic KeratinoSens™ cell line in at least two independent experiment runs. Therefore, the test item can be considered to be a sensitiser in accordance with UN GHS category 1.
- Executive summary:
In the present study the test material was dissolved in DMSO
Based on a molecular weight of 206.29 g/mol a stock solution of 200 mM was prepared.
Based on the stock solution a set of twelve master solutions in 100% solvent was preparedby serial dilution using a constant dilution factor of 1:2. These master solutions were diluted 1:100 in cell culture medium. The following concentration range was tested in the assay:
2000, 1000, 500, 250, 125, 62.5, 31.25, 15.63, 7.81, 3.91, 1.95, 0.98 µM
Cells were incubated with the test item for 48 h at 37°C. After exposurecells were lysed and luciferase activity was assessed by luminescence measurement.
In the firstexperiment, a max luciferase activity (Imax) induction of 6.23 was determined at a test item concentration of 125 µM. The corresponding cell viability was 95.3%. The lowest tested concentration with a significant luciferase induction >1.5 (1.63) was found to be 7.81 µM. The corresponding cell viability was >70% (100.3%).The calculated EC1.5was<1000 µM (6.23 µM).
In the second experiment, a max luciferase activity (Imax) induction of 4.35 was determined at a test item concentration of 125 µM. The corresponding cell viability was 91.0%. The lowest tested concentration with a significant luciferase induction >1.5 (1.76) was found to be 15.63 µM. The corresponding cell viability was >70% (97.6%).The calculated EC1.5was<1000 µM (9.72 µM).
A dose response for luciferase activity induction was observed for each individual run as well as for an overall luciferase activity induction.
Under the condition of this study the test item is therefore considered as sensitiser.
- Endpoint:
- skin sensitisation: in vitro
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- 2017-09-21 to 2018-01-09
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- other: OECD Guidelines for Testing of Chemicals, No. 442E: In Vitro Skin Sensitisation assays addressing the Key Event on activation of dendritic cells on the Adverse Outcome Pathway for Skin Sensitisation”, adopted 09 October 2017
- Qualifier:
- according to guideline
- Guideline:
- other: Human Cell Line Activation Test (h-CLAT) for Skin Sensitisation, DB-ALM Protocol n°158, July 1st, 2015
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany
- Type of study:
- activation of dendritic cells
- Details on the study design:
- The in vitro human cell line activation test (h-CLAT) enables detection of the sensitising potential of a test item by addressing the third molecular key event of the adverse outcome pathway (AOP), namely dendritic cell activation, by quantifying the expression of the cell surface markers CD54 and CD86 in the human monocytic cell line THP-1. The expression of the cell surface markers compared to the respective solvent controls is used to support discrimination between skin sensitisers and non-sensitisers.
- Positive control results:
- The positive control (DNCB) led to an upregulation of the expression of CD54 and CD86 in both experiments. The threshold of 150% for CD86 (259% experiment 1; 373% experiment 2; 254% experiment 3) and 200% for CD54 (244% experiment 1; 375% experiment 2; 335% experiment 3) were clearly exceeded.
- Key result
- Run / experiment:
- other: 1
- Parameter:
- other: relative fluorescence intensity CD86 [%]
- Value:
- 97
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Run / experiment:
- other: 1
- Parameter:
- other: relative fluorescence intensity CD54 [%]
- Value:
- 152
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Run / experiment:
- other: 2
- Parameter:
- other: relative fluorescence intensity CD86 [%]
- Value:
- 158
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Run / experiment:
- other: 2
- Parameter:
- other: relative fluorescence intensity CD54 [%]
- Value:
- 259
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Run / experiment:
- other: 3
- Parameter:
- other: relative fluorescence intensity CD86 [%]
- Value:
- 268
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Run / experiment:
- other: 3
- Parameter:
- other: relative fluorescence intensity CD54 [%]
- Value:
- 761
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Other effects / acceptance of results:
- Acceptance criteria:
The test meets acceptance criteria if:
• the cell viability of the solvent controls is >90%,
• the cell viability of at least four tested doses of the test item in each run is >50%,
• the RFI values of the positive control (DNCB) is ≥150% for CD86 and ≥200% for CD54 at a cell viability of >50%,
• the RFI values of the solvent control is not ≥150% for CD86 and not ≥200% for CD54,
• the MFI ratio of CD86 and CD54 to isotype IgG1 control for the medium and DMSO control, is >105%.
The test mets the acceptance criteria. - Interpretation of results:
- other: The result should be considered in the context of integrated approach such as IATA.
- Conclusions:
- In this study under the given conditions the test item did upregulate the cell surface marker in at least two independent experiment runs. Therefore, the test item is considered to be a skin sensitiser in accordance with UN GHS category 1. However, due to the severe cytotoxic effects at the high test item concentrations it cannot be excluded, that the upregulation of the cell surface marker was caused by the cell stress.
The data generated with this method may be not sufficient to conclude on skin sensitisation potential of chemicals and should be considered in the context of integrated approach such as IATA. - Executive summary:
In the present study the test item was dissolved in DMSO. For the dose finding assay stock solutions with concentrations ranging from 500 mg/mL to 3.91 mg/mL were prepared by a serial dilution of 1:2. Cells were incubated with the test item for 24 h at 37°C. After exposure cells were stained with propidium iodide and cell viability was measured by FACS analysis.
A CV75 of 120.05 ± 17.31 µg/mL was derived in the dose finding assay.
Based on the CV75, the main experiment was performed covering the following concentration steps:
144.07, 120.05, 100.05, 83.37, 69.48, 57.90, 48.25 and 40.21 µg/mL
Cells were incubated with the test item for 24 h at 37°C. After exposure cells were stained and cell surface markers CD54 and CD86 were measured by FACS analysis. Cell viability was assessed in parallel using propidium iodide staining.
Severe cytotoxic effects were observed for the cells treated with the test item. Relative cell viability at the highest test item concentration was reduced to 11.4% (CD86), 10.1% (CD54) and 11.6% (isotype IgG1 control) in the first experiment, to 20.5% (CD86), 20.9% (CD54) and 20.7% (isotype IgG1 control) in the second experiment and 12.0% (CD86), 12.3% (CD54) and 12.0% (isotype IgG1 control) in the second experiment. Severe cytotoxic effects (cell viability < 50%) was observed at the highest two concentrations in experiment 1 and 3 and at the highest concentration only in experiment 2.
In experiment 1 the expression of the cell surface marker CD86 and CD54 was not upregulated above the respective threshold values (150% for CD86 and 200% for CD54).
In experiment 2 the expression of the cell surface marker CD86 was upregulated to 158%. The upregulation above the threshold of 150% was observed starting from a concentration of 83.37 µg/mL, whereas the two highest concentrations showed no upregulation above the threshold. The expression of the cell surface marker CD54 was upregulated to 259%. The upregulation above the threshold of 200% was observed at a concentration of 120.06 µg/mL.
To verify the results a third experiment was performed.
In experiment 3 the expression of the cell surface marker CD54 was upregulated to 258%. The upregulation above the threshold of 200% was observed at a concentration of 100.05 µg/mL. In contrast, the expression of cell surface marker CD86 was not upregulated above the threshold of 150%. Furthermore, for both cell surface marker an upregulation above the respective threshold values was observed at the highest concentration of 144.10 µg/mL, respectively. However, at this concentration the cell viability was < 50%.
Since the cell surface marker clearly exceeded the threshold in two independent experiments the test item considered to be a skin sensitiser. However, due to the severe cytotoxic effects at the high test item concentrations it cannot be excluded, that the upregulation of the cell surface marker was caused by the cell stress.
The positive control (DNCB) led to an upregulation of the expression of CD54 and CD86 in both experiments. The threshold of 150% for CD86 (259% experiment 1; 373% experiment 2; 254% experiment 3) and 200% for CD54 (244% experiment 1; 375% experiment 2; 335% experiment 3) were clearly exceeded.
Referenceopen allclose all
Cysteine and Lysine Values of the Calibration Curve
Sample |
Cysteine Peptide |
Lysine Peptide |
||
Peak Area |
Peptide Concentration [mM] |
Peak Area |
Peptide Concentration [mM] |
|
STD1 |
4835.2368 |
0.5340 |
4196.2754 |
0.5340 |
STD2 |
2314.6648 |
0.2670 |
2135.2583 |
0.2670 |
STD3 |
782.4597 |
0.1335 |
1032.7322 |
0.1335 |
STD4 |
525.4504 |
0.0667 |
516.1820 |
0.0667 |
STD5 |
266.4071 |
0.0334 |
257.2941 |
0.0334 |
STD6 |
135.9798 |
0.0167 |
130.6166 |
0.0167 |
STD7 |
0.0000 |
0.0000 |
0.0000 |
0.0000 |
Depletion of the Cysteine Peptide
Cysteine Peptide |
||||||
Sample |
Peak Area |
Peptide Conc. [mM] |
Peptide Depletion [%] |
Mean Peptide Depletion [%] |
SD of Peptide Depletion [%] |
CV of Peptide Depletion [%] |
Positive Control |
1381.2087 |
0.1629 |
71.03 |
71.19 |
0.17 |
0.24 |
1364.7275 |
0.1611 |
71.37 |
||||
1374.1948 |
0.1621 |
71.18 |
||||
Test Item |
4794.6045 |
0.5384 |
0.00 |
0.06 |
0.09 |
154.04 |
4759.6314 |
0.5345 |
0.17 |
||||
4766.8809 |
0.5353 |
0.01 |
Depletion of the Lysine Peptide
Lysine Peptide |
||||||
Sample |
Peak Area |
Peptide Conc. [mM] |
Peptide Depletion [%] |
Mean Peptide Depletion [%] |
SD of Peptide Depletion [%] |
CV of Peptide Depletion [%] |
Positive Control |
1633.4226 |
0.2075 |
58.76 |
59.19 |
0.48 |
0.81 |
1595.8407 |
0.2028 |
59.71 |
||||
1619.8287 |
0.2058 |
59.10 |
||||
Test Item |
3953.4578 |
0.5018 |
0.18 |
0.26 |
0.15 |
58.26 |
3953.9629 |
0.5018 |
0.17 |
||||
3943.2253 |
0.5005 |
0.44 |
Prediction Model 1
Cysteine 1:10/ Lysine 1:50 Prediction Model 1
Mean Cysteine andLysine PPD |
Reactivity Class |
DPRA Prediction² |
0.00% ≤ PPD ≤ 6.38% |
No or Minimal Reactivity |
Negative |
6.38% < PPD ≤ 22.62% |
Low Reactivity |
Positive |
22.62% < PPD ≤ 42.47% |
Moderate Reactivity |
|
42.47% < PPD ≤ 100% |
High Reactivity |
1 The numbers refer to statistically generated threshold values and are not related to the precision of the measurement.
2 DPRA predictions should be considered in the framework of an IATA.
Prediction Model 2
Cysteine 1:10 Prediction Model
Cysteine PPD |
ReactivityClass |
DPRA Prediction² |
0.00% ≤ PPD ≤ 13.89% |
No or Minimal Reactivity |
Negative |
13.89% < PPD ≤ 23.09% |
Low Reactivity |
Positive |
23.09% < PPD ≤ 98.24% |
Moderate Reactivity |
|
98.24% < PPD ≤ 100% |
High Reactivity |
Categorization of the Test Item
Prediction Model |
Prediction Model 1 |
Prediction Model 2 |
||||
Test Substance |
Mean Peptide Depletion [%] |
Reactivity Category |
Prediction |
Mean Peptide Depletion [%] |
Reactivity Category |
Prediction |
Test Item |
0.16 |
Minimal Reactivity |
no sensitizer |
0.06 |
Minimal Reactivity |
no sensitizer |
Positive Control |
65.19 |
High Reactivity |
sensitizer |
71.19 |
Moderate Reactivity |
sensitizer |
Results of the Cytotoxicity Measurement
|
Concentration [µM] |
Cell Viability [%] |
|||
Experiment 1 |
Experiment 2 |
Mean |
SD |
||
Solvent Control |
- |
100 |
100 |
100 |
0.0 |
Positive Control |
4.00 |
98.9 |
100.0 |
99.5 |
0.7 |
8.00 |
102.4 |
103.0 |
102.7 |
0.4 |
|
16.00 |
104.5 |
107.8 |
106.2 |
2.3 |
|
32.00 |
112.8 |
116.2 |
114.5 |
2.4 |
|
64.00 |
119.1 |
120.9 |
120.0 |
1.3 |
|
Test Item |
0.98 |
96.2 |
99.7 |
97.9 |
2.5 |
1.95 |
93.0 |
87.2 |
90.1 |
4.1 |
|
3.91 |
105.2 |
86.2 |
95.7 |
13.4 |
|
7.81 |
100.3 |
85.8 |
93.1 |
10.2 |
|
15.63 |
110.7 |
97.6 |
104.1 |
9.3 |
|
31.25 |
125.4 |
109.4 |
117.4 |
11.3 |
|
62.50 |
129.1 |
130.6 |
129.8 |
1.0 |
|
125.00 |
95.3 |
91.0 |
93.2 |
3.0 |
|
250.00 |
0.1 |
0.1 |
0.1 |
0.0 |
|
500.00 |
0.3 |
0.1 |
0.2 |
0.1 |
|
1000.00 |
0.1 |
0.1 |
0.1 |
0.0 |
|
2000.00 |
0.4 |
0.4 |
0.4 |
0.0 |
Induction of Luciferase Activity Experiment 1
Experiment 1 |
Concentration [µM] |
Fold Induction |
Significance |
||||
Rep. 1 |
Rep. 2 |
Rep. 3 |
Mean |
SD |
|||
Solvent Control |
- |
1.00 |
1.00 |
1.00 |
1.00 |
0.00 |
|
Positive Control |
4.00 |
1.03 |
1.21 |
1.13 |
1.12 |
0.09 |
|
8.00 |
1.28 |
1.38 |
1.30 |
1.32 |
0.05 |
|
|
16.00 |
1.57 |
1.75 |
1.65 |
1.66 |
0.09 |
* |
|
32.00 |
2.43 |
2.71 |
2.42 |
2.52 |
0.16 |
* |
|
64.00 |
3.82 |
5.53 |
4.29 |
4.55 |
0.88 |
* |
|
Test Item |
0.98 |
1.00 |
1.02 |
1.03 |
1.02 |
0.02 |
|
1.95 |
1.27 |
1.26 |
1.13 |
1.22 |
0.08 |
|
|
3.91 |
1.23 |
1.56 |
1.14 |
1.31 |
0.22 |
|
|
7.81 |
1.65 |
1.74 |
1.50 |
1.63 |
0.12 |
* |
|
15.63 |
1.90 |
2.40 |
1.72 |
2.01 |
0.35 |
* |
|
31.25 |
1.87 |
2.25 |
1.84 |
1.99 |
0.23 |
* |
|
62.50 |
3.04 |
3.24 |
2.37 |
2.88 |
0.45 |
* |
|
125.00 |
5.61 |
8.14 |
4.92 |
6.23 |
1.70 |
* |
|
250.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
|
|
500.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
|
|
1000.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
|
|
2000.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
|
* = significant induction according to Student’s t-test, p<0.05
Induction of Luciferase Activity Experiment 2
Experiment 2 |
Concentration [µM] |
Fold Induction |
Significance |
||||
Rep. 1 |
Rep. 2 |
Rep. 3 |
Mean |
SD |
|||
Solvent Control |
- |
1.00 |
1.00 |
1.00 |
1.00 |
0.00 |
|
Positive Control |
4.00 |
1.27 |
1.22 |
1.00 |
1.16 |
0.15 |
|
8.00 |
1.33 |
1.45 |
1.25 |
1.34 |
0.10 |
|
|
16.00 |
1.76 |
1.66 |
1.37 |
1.60 |
0.20 |
* |
|
32.00 |
2.50 |
2.69 |
1.87 |
2.35 |
0.43 |
* |
|
64.00 |
6.26 |
6.46 |
5.66 |
6.13 |
0.42 |
* |
|
Test Item |
0.98 |
1.04 |
1.14 |
0.79 |
0.99 |
0.18 |
|
1.95 |
1.01 |
1.36 |
0.94 |
1.10 |
0.23 |
|
|
3.91 |
1.19 |
1.28 |
1.21 |
1.23 |
0.05 |
|
|
7.81 |
1.38 |
1.55 |
1.32 |
1.42 |
0.12 |
|
|
15.63 |
1.84 |
2.01 |
1.43 |
1.76 |
0.30 |
* |
|
31.25 |
2.08 |
2.32 |
2.04 |
2.15 |
0.15 |
* |
|
62.50 |
2.58 |
2.90 |
2.14 |
2.54 |
0.38 |
* |
|
125.00 |
4.33 |
4.65 |
4.07 |
4.35 |
0.29 |
* |
|
250.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
|
|
500.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
|
|
1000.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
|
|
2000.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
|
* = significant induction according to Student’s t-test, p<0.05
Induction of Luciferase Activity – Overall Induction
Overall Induction |
Concentration [µM] |
Fold Induction |
Significance |
|||
Experiment 1 |
Experiment 2 |
Mean |
SD |
|||
Solvent Control |
- |
1.00 |
1.00 |
1.00 |
0.00 |
|
Positive Control |
4.00 |
1.12 |
1.16 |
1.14 |
0.03 |
|
8.00 |
1.32 |
1.34 |
1.33 |
0.02 |
|
|
16.00 |
1.66 |
1.60 |
1.63 |
0.04 |
* |
|
32.00 |
2.52 |
2.35 |
2.44 |
0.12 |
* |
|
64.00 |
4.55 |
6.13 |
5.34 |
1.12 |
* |
|
Test Item |
0.98 |
1.02 |
0.99 |
1.00 |
0.02 |
|
1.95 |
1.22 |
1.10 |
1.16 |
0.08 |
|
|
3.91 |
1.31 |
1.23 |
1.27 |
0.06 |
|
|
7.81 |
1.63 |
1.42 |
1.52 |
0.15 |
* |
|
15.63 |
2.01 |
1.76 |
1.88 |
0.17 |
* |
|
31.25 |
1.99 |
2.15 |
2.07 |
0.12 |
* |
|
62.50 |
2.88 |
2.54 |
2.71 |
0.24 |
* |
|
125.00 |
6.23 |
4.35 |
5.29 |
1.32 |
* |
|
250.00 |
0.00 |
0.00 |
0.00 |
0.00 |
|
|
500.00 |
0.00 |
0.00 |
0.00 |
0.00 |
|
|
1000.00 |
0.00 |
0.00 |
0.00 |
0.00 |
|
|
2000.00 |
0.00 |
0.00 |
0.00 |
0.00 |
|
* = significant induction according to Student’s t-test, p<0.05
Additional Parameters
Parameter |
Experiment 1 |
Experiment 2 |
Mean |
SD |
EC1.5[µM] |
6.23 |
9.72 |
7.98 |
2.47 |
Imax |
6.23 |
4.35 |
5.29 |
1.32 |
IC30[µM] |
158.25 |
153.90 |
156.07 |
3.07 |
IC50[µM] |
184.49 |
181.39 |
182.94 |
2.20 |
Acceptance Criteria
Criterion |
Range |
Experiment 1 |
pass/fail |
Experiment 2 |
pass/fail |
CV Solvent Control |
< 20% |
12.7 |
pass |
10.5 |
pass |
No. of positive control concentration steps with significant luciferase activity induction >1.5 |
≥ 1 |
3.0 |
pass |
3.0 |
pass |
EC1.5 PC |
7 < x < 34 µM |
12.29 |
pass |
12.96 |
pass |
Induction PC at 64 µM |
2.00 < x < 8.00 |
4.55 |
pass |
6.13 |
pass |
Historical Data
Acceptance Criterion |
Range |
Mean |
SD |
N |
CV Solvent Control |
< 20% |
11.3 |
3.3 |
41 |
No. of positive control concentration steps with significant luciferase activity induction >1.5 |
≥ 1 |
2.3 |
0.6 |
41 |
EC1.5 PC |
7 < x < 34 µM |
20.4 |
6.7 |
41 |
Induction PC at 64 µM |
2.00 < x < 8.00 |
3.3 |
1.1 |
41 |
Results of the Dose Finding Assay
Sample |
Experiment 1 |
Experiment 2 |
||
Concentration applied [µg/ml] |
Cell Viability [%] |
Concentration applied [µg/ml] |
Cell Viability [%] |
|
Medium Control |
0.00 |
97.30 |
0.00 |
96.80 |
Solvent Control |
0.00 |
96.70 |
0.00 |
97.10 |
Test item |
7.81 |
95.90 |
7.81 |
95.30 |
15.63 |
95.50 |
15.63 |
94.30 |
|
31.25 |
94.90 |
31.25 |
93.20 |
|
62.50 |
91.60 |
62.50 |
92.70 |
|
125.00 |
81.20 |
125.00 |
70.20 |
|
250.00 |
5.40 |
250.00 |
6.50 |
|
500.00 |
4.40 |
500.00 |
2.90 |
|
1000.00 |
9.30 |
1000.00 |
5.40 |
|
Calculated CV75 [µg/mL] |
132.29 |
107.82 |
||
Mean CV75 [µg/mL] |
120.05 |
|||
SD CV 75 [µg/mL] |
17.31 |
CD54 and CD86 Expression Experiment 1
Sample |
Conc. |
Cell Viability [%] |
Mean Fluorescence Intensity |
corrected Mean Fluorescence Intensity |
Relative Flourescence Intensity (RFI) |
Ratio Isotype IgG1 to [%] |
|||||||
CD86 |
CD54 |
Isotype IgG1 |
CD86 |
CD54 |
Isotype IgG1 |
CD86 |
CD54 |
CD86 |
CD54 |
CD86 |
CD54 |
||
Medium Control |
- |
94.4 |
94.6 |
93.7 |
1617 |
1349 |
724 |
893 |
625 |
74 |
90 |
223 |
186 |
Solvent Control |
0.20% |
94.6 |
94.1 |
94.4 |
1865 |
1360 |
663 |
1202 |
697 |
100 |
100 |
281 |
205 |
DNCB |
4.00 |
83.5 |
82.3 |
80.8 |
3736 |
2325 |
627 |
3109 |
1698 |
259 |
244 |
596 |
371 |
Test item |
144.07 |
11.4 |
10.1 |
11.6 |
1448 |
1446 |
918 |
530 |
528 |
44 |
76 |
158 |
158 |
120.06 |
46.4 |
47.6 |
47.3 |
1264 |
1668 |
610 |
654 |
1058 |
54 |
152 |
207 |
273 |
|
100.05 |
66.0 |
66.2 |
66.1 |
1664 |
1710 |
653 |
1011 |
1057 |
84 |
152 |
255 |
262 |
|
83.37 |
77.8 |
79.4 |
79.5 |
1843 |
1588 |
725 |
1118 |
863 |
93 |
124 |
254 |
219 |
|
69.48 |
84.6 |
84.3 |
83.3 |
1911 |
1528 |
769 |
1142 |
759 |
95 |
109 |
249 |
199 |
|
57.90 |
90.6 |
90.1 |
90.6 |
1894 |
1356 |
724 |
1170 |
632 |
97 |
91 |
262 |
187 |
|
48.25 |
90.8 |
91.5 |
90.8 |
1758 |
1279 |
714 |
1044 |
565 |
87 |
81 |
246 |
179 |
|
40.21 |
90.4 |
91.3 |
89.9 |
1745 |
1283 |
705 |
1040 |
578 |
87 |
83 |
248 |
182 |
CD54 and CD86 Expression Experiment 2
Sample |
Conc. |
Cell Viability [%] |
Mean Fluorescence Intensity |
corrected Mean Fluorescence Intensity |
Relative Flourescence Intensity (RFI) |
Ratio Isotype IgG1 to [%] |
|||||||
CD86 |
CD54 |
Isotype IgG1 |
CD86 |
CD54 |
Isotype IgG1 |
CD86 |
CD54 |
CD86 |
CD54 |
C86 |
CD54 |
||
Medium Control |
- |
96.4 |
95.9 |
96.1 |
1594 |
1277 |
637 |
957 |
640 |
94 |
101 |
250 |
200 |
Solvent Control |
0.20% |
96.7 |
96.7 |
96.6 |
1693 |
1311 |
679 |
1014 |
632 |
100 |
100 |
249 |
193 |
DNCB |
4.0 |
81.2 |
81.4 |
80.6 |
4482 |
3075 |
704 |
3778 |
2371 |
373 |
375 |
637 |
437 |
Test item |
144.07 |
20.5 |
20.9 |
20.7 |
1419 |
1427 |
710 |
709 |
717 |
70 |
113 |
200 |
201 |
120.06 |
64.9 |
65.2 |
66.6 |
2312 |
2447 |
812 |
1500 |
1635 |
148 |
259 |
285 |
301 |
|
100.05 |
78.5 |
78.9 |
78.8 |
2399 |
1890 |
799 |
1600 |
1091 |
158 |
173 |
300 |
237 |
|
83.37 |
79.6 |
81.2 |
81.0 |
2373 |
1815 |
767 |
1606 |
1048 |
158 |
166 |
309 |
237 |
|
69.48 |
86.5 |
86.0 |
86.1 |
2192 |
1722 |
811 |
1381 |
911 |
136 |
144 |
270 |
212 |
|
57.90 |
88.0 |
88.3 |
87.6 |
2113 |
1684 |
852 |
1261 |
832 |
124 |
132 |
248 |
198 |
|
48.25 |
89.3 |
90.0 |
88.9 |
2060 |
1610 |
835 |
1225 |
775 |
121 |
123 |
247 |
193 |
|
40.21 |
91.6 |
91.1 |
91.4 |
1984 |
1647 |
907 |
1077 |
740 |
106 |
117 |
219 |
182 |
CD54 and CD86 Expression Experiment 3
Sample |
Conc. |
Cell Viability [%] |
Mean Fluorescence Intensity |
corrected Mean Fluorescence Intensity |
Relative Flourescence Intensity (RFI) |
Ratio Isotype IgG1 to [%] |
|||||||
CD86 |
CD54 |
Isotype IgG1 |
CD86 |
CD54 |
Isotype IgG1 |
CD86 |
CD54 |
CD86 |
CD54 |
C86 |
CD54 |
||
Medium Control |
- |
95.0 |
95.3 |
95.0 |
2630 |
1252 |
784 |
1846 |
468 |
70 |
93 |
335 |
160 |
Solvent Control |
0.20% |
95.0 |
94.8 |
94.9 |
3417 |
1275 |
774 |
2643 |
501 |
100 |
100 |
441 |
165 |
DNCB |
4.0 |
83.0 |
83.9 |
82.7 |
7420 |
2393 |
714 |
6706 |
1679 |
254 |
335 |
1039 |
335 |
Test item |
144.1 |
12.0 |
12.3 |
12.0 |
9305 |
6023 |
2211 |
7094 |
3812 |
268 |
761 |
421 |
272 |
120.06 |
30.2 |
31.2 |
29.6 |
2927 |
1586 |
640 |
2287 |
946 |
87 |
189 |
457 |
248 |
|
100.05 |
61.2 |
62.4 |
62.6 |
3158 |
1959 |
668 |
2490 |
1291 |
94 |
258 |
473 |
293 |
|
83.37 |
74.7 |
76.0 |
75.9 |
3371 |
1616 |
753 |
2618 |
863 |
99 |
172 |
448 |
215 |
|
69.48 |
83.8 |
83.4 |
82.8 |
3340 |
1476 |
764 |
2576 |
712 |
97 |
142 |
437 |
193 |
|
57.90 |
86.2 |
86.8 |
86.0 |
2952 |
1429 |
780 |
2172 |
649 |
82 |
130 |
378 |
183 |
|
48.25 |
87.4 |
87.6 |
87.8 |
2918 |
1374 |
873 |
2045 |
501 |
77 |
100 |
334 |
157 |
|
40.21 |
89.0 |
89.7 |
88.9 |
3056 |
1332 |
803 |
2253 |
529 |
85 |
106 |
381 |
166 |
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed (sensitising)
Respiratory sensitisation
Endpoint conclusion
- Endpoint conclusion:
- no study available
Justification for classification or non-classification
Based on the provided information there is sufficient evidence that the substance needs to classified as skin sensitiser class 1 according to the EU Regulation (EC) No 1272/2008 on Classification, Labelling and Packaging of Substances and Mixtures.
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.