Registration Dossier
Registration Dossier
Data platform availability banner - registered substances factsheets
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Diss Factsheets
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 821-885-7 | CAS number: 1820028-80-5
- 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
Endpoint summary
Administrative data
Description of key information
Initially, skin sensitisation was studied in vitro using the three key events, namely the DPRA, the Keratinosens and the hClat Assay.
In the DPRA assay, no indication for peptide depletion was shown. However, due to the observed precipitation of the test material, the prediction model does not apply. In the Keratinosens assay, an increase of the luciferase activity was shown in the transgenic cells, indicating potential for skin sensitisation.
Therefore, the third key event, the hClat assay, was started to verify this result. In this OECD 442E study, there was no upregulation of the expression of the relevant cell surface markers CD86 and CD54 observed, however, the logP of the test material exceeds the threshold of 3.5. Thus, according to OECD 442E, negative results must be considered as inconclusive.
Given the fact that no firm conclusion based on on vitro data for skin sensitisation could be drawn, a further in vivo test was conducted.
In this assay the OECD GLP criteria were met and the methods applied are fully compliant with OECD TG 429. In this local lymph node assay test item concentrations of 5, 10, and 25% (w/w) were applied. The highest concentration tested was the highest concentration that could technically be achieved. The analysis of the SI values showed that the test item is a skin sensitiser. An EC3 value could not be derived since all SI values obtained were above the threshold index of 3.
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:
- key study
- Study period:
- 2019-02-06 to 2019-02-14
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: GLP 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:
- other: (in chemico) reactivity against synthetic peptides with a thiol or amino group
- 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 cysteine peptide. The mean depletion of the cysteine peptide was 69.94%.
- Key result
- Run / experiment:
- other: cysteine run
- Parameter:
- other: mean peptide depletion [%]
- Value:
- 5.07
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Run / experiment:
- other: lysine run
- Parameter:
- other: mean peptide depletion [%]
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- not determinable
- 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. - Conclusions:
- The samples containing lysine peptide could not be analysed due to strong precipitation. In this study under the given conditions the test item showed minimal reactivity towards the cysteine peptide. Due to the observed precipitation in the cysteine experiment a test item concentration of 100 mM cannot be guaranteed and a prediction cannot be made.
The data generated with this test should be considered in the context of integrated approached such as IATA, combining the result with other complementary information, e.g. derived from in vitro assays addressing other key events of the skin sensitisation AOP. - Endpoint:
- skin sensitisation: in vitro
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2019-02-15 to 2019-03-15
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: GLP 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 (3.48 (experiment 1); 2.39 (experiment 2)).
- Key result
- Run / experiment:
- other: 1
- Parameter:
- other: luciferase activity
- Value:
- 4.09
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: Concentration: 3.91 µM
- Key result
- Run / experiment:
- other: 1
- Parameter:
- other: cell viability [%]
- Value:
- 110.3
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Run / experiment:
- other: 1
- Parameter:
- other: EC1.5 [µM]
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- not determinable
- Key result
- Run / experiment:
- other: 2
- Parameter:
- other: luciferase activity
- Value:
- 3.16
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: Concentration: 1.95 µM
- Key result
- Run / experiment:
- other: 2
- Parameter:
- other: cell viability [%]
- Value:
- 100.9
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Run / experiment:
- other: 2
- Parameter:
- other: EC1.5 [µM]
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- not determinable
- 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. - 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 as sensitiser.
The data generated with this test should be considered in the context of integrated approached such as IATA, combining the result with other complementary information, e.g. derived from in vitro assays addressing other key events of the skin sensitisation AOP. - Endpoint:
- skin sensitisation: in vitro
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2019-06-27 to 2019-08-01
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: GLP 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 25 June 2018
- 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 (244% experiment 1; 366% experiment 2) and
200% for CD54 (298% experiment 1; 459% experiment 2) were clearly exceeded. - Key result
- Run / experiment:
- other: 1
- Parameter:
- other: relative fluorescence intensity CD86 [%]
- Value:
- 120
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: Concentration: 9.78 µg/mL
- Key result
- Run / experiment:
- other: 1
- Parameter:
- other: relative fluorescence intensity CD54 [%]
- Value:
- 126
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: Concentration: 5.66 µg/mL
- Key result
- Run / experiment:
- other: 2
- Parameter:
- other: relative fluorescence intensity CD86 [%]
- Value:
- 93
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: Concentration: 9.78 µg/mL
- Key result
- Run / experiment:
- other: 2
- Parameter:
- other: relative fluorescence intensity CD54 [%]
- Value:
- 128
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: Concentration: 6.79 µg/mL
- 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. - Conclusions:
- In this study under the given conditions the test item did not upregulate the expression of the cell surface markers in at least two independent experiment runs.
Therefore, 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. - Endpoint:
- skin sensitisation: in vivo (LLNA)
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- Nov 15, 2019 - Jan 23, 2020
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of study:
- mouse local lymph node assay (LLNA)
- Species:
- mouse
- Strain:
- CBA
- Sex:
- female
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source: Envigo RMS B.V., Inc Postbus 6174 5960 AD Horst / The Netherlands
- Age at study initiation: Pre-test test: 12 - 13 weeks
Main Test: 9-9 weeks
- Weight at study initiation: Pre-test and Main test: 16.9 - 21.2 g
- Housing: grouped per dose
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 + 2°C
- Humidity (%): 22-65%
- Photoperiod (hrs dark / hrs light): 12/12
IN-LIFE DATES: From: day 1 To: day 6 - Vehicle:
- propylene glycol
- Concentration:
- 5, 10, and 25 (w/w)
- No. of animals per dose:
- 5
- Details on study design:
- RANGE FINDING TESTS:
- Compound solubility: 25% in MEK
- Irritation: No
- Lymph node proliferation response: -
MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: OECD 429
- Criteria used to consider a positive response: Stimulation index > 3 - Positive control substance(s):
- hexyl cinnamic aldehyde (CAS No 101-86-0)
- Statistics:
- Standard statistical methods have been applied for data processing.
- Positive control results:
- Conc. SI
0%: 1.0
5% 7.7
10% 7.5
25% 11.7 - Key result
- Parameter:
- SI
- Value:
- 7.7
- Test group / Remarks:
- Test Group: 5% in MEK
- Key result
- Parameter:
- SI
- Value:
- 7.5
- Test group / Remarks:
- Test Group: 10% in MEK
- Key result
- Parameter:
- SI
- Value:
- 11.7
- Test group / Remarks:
- Test Group: 25% in MEK
- Cellular proliferation data / Observations:
-
EC3 CALCULATION : EC3 = (a-c) [(3-d)/(b-d)] + c; a,b,c,d = co-ordinates of the two pairs of data lying immediately above and below the S.I. value of 3 on the LLNA dose response plot
CLINICAL OBSERVATIONS: No deaths occurred during the study period. No symptoms of local skin irritation at the ears of the animals and no signs of systemic toxicity were observed during the study period.
BODY WEIGHTS: The body weight of the animals, recorded prior to the first application and prior to treatment with 3HTdR, was within the range commonly recorded for animals of this strain and age. - Interpretation of results:
- Category 1B (indication of skin sensitising potential) based on GHS criteria
- Conclusions:
- The test item was found to be a skin sensitiser. An EC3 value could not be derived since all S.I. values obtained were above the threshold index of 3.
- Executive summary:
The information for this endpoint study record was obtained from an experimental study. The OECD GLP criteria were met and the methods applied are fully compliant with OECD TG 429.
For this purpose a local lymph node assay was performed using test item concentrations of 5, 10, and 25% (w/w). The highest concentration tested was the highest concentration that could technically be achieved.
The test item was found to be a skin sensitiser. An EC3 value could not be derived since all S.I. values obtained were above the threshold index of 3.
Referenceopen allclose all
Cysteine Values of the Calibration Curve
Sample |
Cysteine Peptide |
|
Peak Area at 220 nm |
Peptide Concentration [mM] |
|
STD1 |
17.4940 |
0.5340 |
STD2 |
8.7090 |
0.2670 |
STD3 |
4.3730 |
0.1335 |
STD4 |
2.1750 |
0.0667 |
STD5 |
1.0820 |
0.0334 |
STD6 |
0.5670 |
0.0167 |
STD7 |
0.0000 |
0.0000 |
Depletion of the Cysteine Peptide
Cysteine Peptide |
||||||
Sample |
Peak Area at 220 nm |
Peptide Concentration [mM] |
Peptide Depletion [%] |
Mean Peptide Depletion [%] |
SD of Peptide Depletion [%] |
CV of Peptide Depletion [%] |
Positive Control |
4.8780 |
0.1491 |
70.09 |
69.94 |
0.43 |
0.62 |
4.8480 |
0.1482 |
70.27 |
||||
4.9820 |
0.1523 |
69.45 |
||||
Test Item |
15.6390 |
0.4778 |
4.26 |
5.07 |
0.74 |
14.53 |
15.4770 |
0.4728 |
5.25 |
||||
15.4040 |
0.4706 |
5.70 |
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 |
- |
- |
- |
5.07* |
Minimal reactivity* |
negative* |
Positive Control |
- |
- |
- |
69.94 |
Moderate Reactivity |
positive |
* only for the sake of completeness
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 |
95.7 |
99.3 |
97.5 |
2.5 |
8.00 |
93.9 |
97.3 |
95.6 |
2.4 |
|
16.00 |
95.5 |
99.9 |
97.7 |
3.1 |
|
32.00 |
85.0 |
87.4 |
86.2 |
1.7 |
|
64.00 |
73.7 |
100.1 |
86.9 |
18.7 |
|
Test Item |
0.98 |
91.4 |
116.8 |
104.1 |
17.9 |
1.95 |
107.4 |
100.9 |
104.2 |
4.6 |
|
3.91 |
110.3 |
101.8 |
106.0 |
6.0 |
|
7.81 |
93.9 |
109.5 |
101.7 |
11.1 |
|
15.63 |
85.5 |
27.0 |
56.3 |
41.4 |
|
31.25 |
1.2 |
0.2 |
0.7 |
0.6 |
|
62.50 |
0.5 |
0.9 |
0.7 |
0.3 |
|
125.00 |
0.2 |
0.6 |
0.4 |
0.3 |
|
250.00 |
0.2 |
1.4 |
0.8 |
0.9 |
|
500.00 |
2.3 |
2.7 |
2.5 |
0.3 |
|
1000.00 |
2.6 |
4.1 |
3.3 |
1.1 |
|
2000.00 |
0.3 |
0.2 |
0.3 |
0.1 |
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 |
1.13 |
1.19 |
1.22 |
1.18 |
0.05 |
|
8.00 |
1.29 |
1.57 |
1.40 |
1.42 |
0.14 |
|
|
16.00 |
1.42 |
1.56 |
1.40 |
1.46 |
0.09 |
|
|
32.00 |
1.32 |
1.97 |
2.12 |
1.81 |
0.43 |
* |
|
64.00 |
3.01 |
3.42 |
4.01 |
3.48 |
0.51 |
* |
|
Test Item |
0.98 |
2.38 |
2.28 |
2.32 |
2.33 |
0.05 |
* |
1.95 |
2.93 |
3.21 |
2.84 |
2.99 |
0.19 |
* |
|
3.91 |
3.98 |
4.48 |
3.80 |
4.09 |
0.35 |
* |
|
7.81 |
3.75 |
4.15 |
3.77 |
3.89 |
0.23 |
* |
|
15.63 |
2.04 |
2.32 |
2.05 |
2.14 |
0.16 |
* |
|
31.25 |
11.99 |
11.96 |
15.08 |
13.01 |
1.79 |
* |
|
62.50 |
0.00 |
0.00 |
0.01 |
0.00 |
0.00 |
|
|
125.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
|
|
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 |
1.03 |
0.98 |
0.97 |
0.99 |
0.04 |
|
8.00 |
1.15 |
1.08 |
1.01 |
1.08 |
0.07 |
|
|
16.00 |
1.24 |
1.16 |
1.20 |
1.20 |
0.04 |
|
|
32.00 |
1.64 |
1.67 |
1.45 |
1.59 |
0.12 |
* |
|
64.00 |
2.38 |
2.47 |
2.32 |
2.39 |
0.08 |
* |
|
Test Item |
0.98 |
2.46 |
2.27 |
2.49 |
2.41 |
0.12 |
* |
1.95 |
3.18 |
3.14 |
3.17 |
3.16 |
0.02 |
* |
|
3.91 |
3.32 |
3.22 |
2.82 |
3.12 |
0.26 |
* |
|
7.81 |
1.74 |
1.86 |
1.73 |
1.78 |
0.08 |
* |
|
15.63 |
6.81 |
10.06 |
7.39 |
8.09 |
1.73 |
* |
|
31.25 |
0.07 |
0.08 |
0.05 |
0.06 |
0.01 |
|
|
62.50 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
|
|
125.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
|
|
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
|
Concentration [µM] |
Fold Induction |
|||
Experiment 1 |
Experiment 2 |
Mean |
SD |
||
Solvent Control |
- |
1.00 |
1.00 |
1.00 |
0.00 |
Positive Control |
4.00 |
1.18 |
0.99 |
1.09 |
0.13 |
8.00 |
1.42 |
1.08 |
1.25 |
0.24 |
|
16.00 |
1.46 |
1.20 |
1.33 |
0.18 |
|
32.00 |
1.81 |
1.59 |
1.70 |
0.15 |
|
64.00 |
3.48 |
2.39 |
2.93 |
0.77 |
|
Test Item |
0.98 |
2.33 |
2.41 |
2.37 |
0.06 |
1.95 |
2.99 |
3.16 |
3.08 |
0.12 |
|
3.91 |
4.09 |
3.12 |
3.60 |
0.69 |
|
7.81 |
3.89 |
1.78 |
2.83 |
1.49 |
|
15.63 |
2.14 |
8.09 |
5.11 |
4.21 |
|
31.25 |
13.01 |
0.06 |
6.54 |
9.15 |
|
62.50 |
0.00 |
0.00 |
0.00 |
0.00 |
|
125.00 |
0.00 |
0.00 |
0.00 |
0.00 |
|
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] |
n.a. |
n.a. |
n.a. |
n.a. |
Imax |
13.01 |
8.09 |
10.55 |
3.48 |
IC30[µM] |
18.50 |
11.55 |
15.03 |
4.91 |
IC50[µM] |
22.20 |
13.45 |
17.83 |
6.19 |
n.a.: not applicable
Acceptance Criteria
Criterion |
Range |
Experiment 1 |
pass/fail |
Experiment 2 |
pass/fail |
CV Solvent Control |
< 20% |
7.9 |
pass |
12.3 |
pass |
No. of positive control concentration steps with significant luciferase activity induction >1.5 |
≥ 1 |
2.0 |
pass |
2.0 |
pass |
EC1.5 PC |
± 2 x SD of historical mean |
17.73 |
pass |
28.31 |
pass |
Induction PC at 64 µM |
2 .00 < x < 8.00 |
3.48 |
pass |
2.39 |
pass |
Historical Data
Acceptance Criterion |
Range |
Mean |
SD |
N |
CV Solvent Control |
< 20% |
11.6 |
3.5 |
96 |
No. of positive control concentration steps with significant luciferase activity induction >1.5 |
≥ 1 |
2.4 |
0.6 |
96 |
EC1.5 PC |
7 < x < 34 µM |
18.5 |
6.0 |
96 |
Induction PC at 64 µM |
2.00 < x < 8.00 |
3.8 |
1.5 |
96 |
Experiment 1:
Sample |
Conc. |
Cell Viability [%] |
Mean Fluorescence Intensity |
corrected Mean Fluorescence Intensity |
Relative Fluorescence Intensity (RFI) |
Ratio Isotype IgG1 to [%] |
|||||||
CD86 |
CD54 |
Isotype IgG1 |
CD86 |
CD54 |
Isotype IgG1 |
CD86 |
CD54 |
CD86 |
CD54 |
CD86 |
CD54 |
||
Medium Control |
- |
95.1 |
94.3 |
94.6 |
1982 |
1209 |
752 |
1230 |
457 |
86 |
94 |
264 |
161 |
Solvent Control 2 (THF) |
0.20% |
94.9 |
95.3 |
95.3 |
1786 |
1281 |
808 |
978 |
473 |
100 |
100 |
221 |
159 |
Solvent Control 1 (DMSO) |
0.20% |
93.6 |
94 |
92.8 |
2206 |
1268 |
780 |
1426 |
488 |
100 |
100 |
283 |
163 |
DNCB |
4.00 |
85.7 |
86 |
84.3 |
4288 |
2264 |
808 |
3480 |
1456 |
244 |
298 |
531 |
280 |
Test item |
11.74 |
33.6 |
30.7 |
31.9 |
2102 |
1238 |
941 |
1161 |
297 |
119 |
63 |
223 |
132 |
9.78 |
38.1 |
36.9 |
38.5 |
2200 |
1407 |
1031 |
1169 |
376 |
120 |
79 |
213 |
136 |
|
8.15 |
66.4 |
66 |
64.1 |
1586 |
1495 |
977 |
609 |
518 |
62 |
110 |
162 |
153 |
|
6.79 |
84.6 |
85.3 |
85.1 |
1473 |
1517 |
947 |
526 |
570 |
54 |
121 |
156 |
160 |
|
5.66 |
90.6 |
90 |
90.1 |
1594 |
1446 |
849 |
745 |
597 |
76 |
126 |
188 |
170 |
|
4.72 |
92.2 |
91.8 |
91.2 |
1572 |
1365 |
890 |
682 |
475 |
70 |
100 |
177 |
153 |
|
3.93 |
92.6 |
92.6 |
92.6 |
1466 |
1305 |
878 |
588 |
427 |
60 |
90 |
167 |
149 |
|
3.28 |
93.6 |
93.4 |
94.3 |
1552 |
1341 |
892 |
660 |
449 |
67 |
95 |
174 |
150 |
Experiment 2:
Sample |
Conc. |
Cell Viability [%] |
Mean Fluorescence Intensity |
corrected Mean Fluorescence Intensity |
Relative Fluorescence Intensity (RFI) |
Ratio Isotype IgG1 to [%] |
|||||||
CD86 |
CD54 |
Isotype IgG1 |
CD86 |
CD54 |
Isotype IgG1 |
CD86 |
CD54 |
CD86 |
CD54 |
CD86 |
CD54 |
||
Medium Control |
- |
96.8 |
96.2 |
96 |
1140 |
882 |
682 |
458 |
200 |
112 |
102 |
167 |
129 |
Solvent Control 2 (THF) |
0.20% |
96 |
95.9 |
95.9 |
1153 |
905 |
704 |
449 |
201 |
100 |
100 |
164 |
129 |
Solvent Control 1 (DMSO) |
0.20% |
95.5 |
95.7 |
95.9 |
1105 |
892 |
696 |
409 |
196 |
100 |
100 |
159 |
128 |
DNCB |
4.00 |
85.7 |
85.3 |
83.8 |
2287 |
1691 |
792 |
1495 |
899 |
366 |
459 |
289 |
214 |
Test item |
11.74 |
55.1 |
54.9 |
55.8 |
1241 |
1057 |
869 |
372 |
188 |
83 |
94 |
143 |
122 |
9.78 |
47.5 |
46.1 |
47.1 |
1239 |
946 |
821 |
418 |
125 |
93 |
62 |
151 |
115 |
|
8.15 |
75.7 |
75.5 |
75.8 |
1095 |
1107 |
861 |
234 |
246 |
52 |
122 |
127 |
129 |
|
6.79 |
91.7 |
91.4 |
90 |
1010 |
1122 |
865 |
145 |
257 |
32 |
128 |
117 |
130 |
|
5.66 |
92.6 |
93.9 |
93.9 |
848 |
947 |
781 |
67 |
166 |
15 |
83 |
109 |
121 |
|
4.72 |
92.2 |
93.7 |
90.5 |
943 |
989 |
821 |
122 |
168 |
27 |
84 |
115 |
120 |
|
3.93 |
93.3 |
93.2 |
93.5 |
926 |
935 |
730 |
196 |
205 |
44 |
102 |
127 |
128 |
|
3.28 |
95.3 |
94.2 |
94.4 |
949 |
967 |
763 |
186 |
204 |
41 |
101 |
124 |
127 |
Calculation of Stimulation Indices per Dose Group:
Test item concentration |
Group Calculation |
||
Mean DPM per |
SD |
S.I. |
|
Vehicle Control Group (MEK) |
1214.4 |
351.3 |
1.0 |
5% Test Item |
9299.6 |
4525.9 |
7.7S |
10% Test Item |
9048.8 |
2631.7 |
7.5S |
25% Test Item |
14237.6 |
6932.5 |
11.7 S |
a) Mean DPM/animal was determined by dividing the sum of the measured values from lymph nodes of all animals within a group by the number of animals in that group (5 animals)
S statistically significant (p<0.05) vs. concurrent vehicle control
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 need to classified according to the EU Regulation (EC) No 1272/2008 on Classification, Labelling and Packaging of Substances and Mixtures. As the EC3 value could not be determined and may be below 2 % Category 1B is justified.
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.