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EC number: 485-320-2 | CAS number: 221667-31-8
- 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
Skin sensitisation
Administrative data
- Endpoint:
- skin sensitisation: in vitro
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 04 - 14 Feb 2020
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study with acceptable restrictions
- Remarks:
- non-GLP
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 020
- Report date:
- 2020
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 442D (In Vitro Skin Sensitisation: ARE-Nrf2 luciferase KeratinoSens™ test method)
- Version / remarks:
- June 2018
- Deviations:
- no
- GLP compliance:
- no
- Remarks:
- The study is supplementary to the main study, which is fully reliable on its own. The study is not intended to replace the already existing in vivo study for skin sensitization, only to substantiate the data package.
- Type of study:
- ARE-Nrf2 luciferase LuSens test method
Test material
- Reference substance name:
- -
- EC Number:
- 485-320-2
- EC Name:
- -
- Cas Number:
- 221667-31-8
- Molecular formula:
- C18H18N205S
- IUPAC Name:
- N-[4-(cyclopropylcarbamoyl)benzenesulfonyl]-2-methoxybenzamide
Constituent 1
In vitro test system
- Details of test system:
- Lusens transgenic cell line [442D]
- Details on the study design:
- CELL LINE
- The LuSens cell-line is an immortalised adherent cell line derived from HaCaT human keratinocytes stably transfected with a selectable plasmid. The plasmid contains a luciferase gene (reporter gene) which is under transcriptional control of an antioxidant response element (ARE) of the rat NQO1 gene. Genes dependent on the ARE such as NQO1 are known to be upregulated by contact sensitisers.
- The HaCaT human keratinocytes (provided by RWTH, Aachen, Germany) were genetically modified at the Institute of Anatomy and Cell Biology of the RWTH, Aachen (laboratory of Wruck). The LuSens cells were obtained by BASF and in 2019 a contract services agreement was established between ICCR-Roßdorf GmbH (Licensee) and Promega.
- The passage numbers of the used LuSens cells were 9 in the cytotoxicity test and 11 in the LuSens test for the main experiments 1 and 2, respectively.
PREPARATION OF TEST SOLUTIONS
- Preparation of the test chemical stock solution: on the day of the experiment (immediately before treatment) the test-item was dissolved or stably dispersed/suspended in DMSO to prepare a stock solution with a concentration of 200 mM in accordance to the OECD Guideline 442D.
- Preparation of the test chemical serial dilutions: for the MTT test (dose finding assay) twelve concentrations of the test item were analysed. Therefore, dilutions were prepared by 1:2 serial dilutions from the highest soluble/dispersible concentration.
Six test item concentrations were tested in the main experiments. The highest concentration used was CV75 × 1.2. Five further test item dilutions were prepared by serial dilution with a dilution factor of 1.2.
- Preparation of the positive controls: EGDMA (final concentration 120 µM), CAS 97-90-5, purity: ≥ 97.5%. The solvent for the positive control was Treatment Medium including 1% (v/v) DMSO
- Preparation of the solvent control: DMSO (final concentration 1% (v/v) in Treatment Medium), purity: ≥ 99%.
- Preparation of the negative control: Lactic acid (final concentration 5000 µM), CAS 50-21-5, purity: ~ 90%. The solvent for the negative control was Treatment Medium including 1% (v/v) DMSO.
CULTURE MEDIUM
DMEM with supplements listed below was used to culture the cells during the assay. The culture medium was warmed to room temperature just before use.
- Cultivation Medium: DMEM culture medium with Penicillin/Streptomycin (1% v/v), FBS (10% v/v) and puromycin (0.005% v/v)
- Seeding Medium: DMEM culture medium with FBS (10% v/v)
- Treatment Medium: DMEM culture medium with FBS (1% v/v)
MTT DOSE RANGE FINDING ASSAY
- One cytotoxicity experiment (dose finding assay) was performed to obtain a CV75, if possible.
- The MTT working solution consisted of two components, the MTT stock solution (5 mg/mL MTT in Ca2+/Mg2+ free DPBS) and Treatment Medium. Both components were mixed right before application at a ratio of 1:10.
- For the treatment of the cells in the dose finding assay a stock solution of the test item and the controls were prepared. The test item was dissolved in the solvent and 1:2 serially diluted in the solvent to obtain the desired test item concentrations (twelve concentrations).
- The solvent (twelve replicates), the positive (two replicates) and the negative (three replicates) controls as well as the test item concentrations (each three replicates) were subsequently diluted 1:25 in Treatment Medium. The final test-item concentrations in the test medium were as follows:
0.98, 1.95, 3.91, 7.81, 15.6, 31.3, 62.5, 125, 250, 500, 1000 and 2000 µM
- 24 hours ± 30 minutes after seeding of the cells, the Seeding Medium were removed from the wells. Thereafter, 150 µL Treatment Medium were added per well and 50 µL of the test item dilutions, the solvent, negative and positive controls (1:25 dilution in Treatment Medium) and
the medium control (six replicates) were added to the wells, respectively. At the end of the incubation period of 48 ± 1 hours under standard cell culture conditions, the cell cultures were microscopically evaluated for morphological alterations, precipitation or phase separation.
- At the end of the incubation period, Treatment Medium was removed from the wells and the cells were washed at least twice with 200 µL DPBS including Ca2+/Mg2+. Thereafter, 200 µL of the MTT working solution were added to each treatment well and the cells were incubated for 3
hours ± 30 minutes under standard cell culture conditions. After rinsing the MTT working solution, the cells of each well were treated with 100 µL MTT lysis agent (Isopropanol with 0.04 N HCl) for at least 30 minutes, while gently shaking. Thereafter the microplate was transferred to a microplate reader (Multimode Reader (TriStar2 LB 942) by Berthold Technologies GmbH Co KG, Germany) equipped with a 570 nm filter to measure the absorbance (reference wavelength 690 nm).
- Cell viability of the test-item treated cells was calculated relative to the solvent control.
- The MTT assay was considered to be acceptable if it meets the following criteria: mean absorbance of the solvent control is ≥ 0.2.
MAIN EXPERIMENT (LuSens and MTT):
- Number of replicates: test item (three replicates per concentration, solvent control (twenty-four replicates), positive control (five replicates) and negative control (six replicates).
- Number of repetitions: 2 independent experiments.
- Test chemical concentrations: 321, 385, 462, 554, 665 and 798 µM
- Preparation and Seeding of Cells conditions: between 4 and 6 x 10E+5 or 6 and 8 x 10E+5 cells were seeded in 15 mL Cultivation Medium and pre-cultured at least twice a week in culture flasks (80 – 90% confluent). The cell density between approximately 80 and 90% should not be exceeded.
After microscopic assessment of the cells, the cells were washed at least twice with 10 mL Ca2+/Mg2+ free DPBS including EDTA. Thereafter, the cells were trypsinated with 1 to 2 mL 0.05% Trypsin/EDTA for approximately 5 minutes at 37 ± 1.5 °C and 5.0 ± 0.5 % CO2. The cells were resuspended in 10 mL Cultivation Medium to neutralise the trypsin.
For seeding of the cells, the Cultivation Medium was removed and the cells were transferred into Seeding Medium. Each treatment well of a 96 well microtiter plate was seeded with 100 µL cell suspension (9000 to 11000 LuSens cells per well), except the well of the blank control. The cells were incubated for 24 hours ± 30 minutes under standard cell culture conditions.
- Incubation conditions: 37 ± 1.5 °C and 5.0 ± 0.5 % CO2
- Application procedure: after incubation of the LuSens cells, Seeding Medium was removed and 150 µL of Treatment Medium was distributed in each well. Thereafter, 50 µL of the test item and control dilutions and the medium control (twelve replicates) were added into the corresponding wells.
- Exposure time: 48 ± 1 hours
LUCIFERASE ACTIVITY MEASUREMENTS
- The Steady-Glo® Mix was be prepared by adding Steady-Glo® buffer to one bottle of SteadyGlo® Substrate and mixing by inversion until the substrate was dissolved. The Steady-Glo® working solution was prepared by mixing one part of DPBS (without Ca2+/Mg2+) with one part
of Steady-Glo®-Mix.
At the end of the incubation period, the Treatment Medium was removed from the wells and the cells were washed at least twice with 200 µL DPBS including Ca2+/Mg2+. Thereafter, 200 µL of the Steady-Glo® working solution was added in each well. After slowly shaking of the microtiter
plate for at least 10 min in the dark, the plate was transferred to a microplate reader and the luminescence was measured for 2 seconds per well.
- Luminometer: Multimode Reader (TriStar2 LB 942) by Berthold Technologies GmbH Co KG, Germany.
MTT VIABILITY ASSAY
- The treatment of the cells with the MTT labelling mixture and the measurement of the cell viability for each main experiment was performed as described above in the dose range finder experiment.
PREDICTION MODEL
If the luciferase induction is ≥ 1.5 fold and statistically significant compared to the solvent control in at least 2 consecutive non-cytotoxic tested concentrations (cell viability ≥ 70%) and at least 3 tested concentrations are non-cytotoxic, the main experiment of the LuSens prediction is considered positive.
If these conditions are met in 2 of 2 or in 2 of 3 main experiments, the LuSens prediction is considered positive, otherwise the LuSens prediction is considered negative.
A negative result obtained with a test item that does not form a stable dispersion and was not tested up to 2000 μM (or 2000 μg/mL for test items with no defined MW) and for which no cytotoxicity is observed in any of the tested concentration should be considered as inconclusive (OECD 442E).
If no clear dose-response curve or a biphasic dose-response curve is observed, the experiment should be repeated to verify whether this is specific to the test item or due to an experimental artefact. If the biphasic response (i.e. when the threshold of 1.5 is crossed twice) is reproducible in an independent verification experiment, the lower concentration of a ≥ 1.5 induction should be reported (i.e. when the threshold of 1.5 is crossed the first time).
Mono constituent test items with a Log Pow > 7 may be insoluble in the culture medium. However, if the test item is soluble or can be stably dispersed/suspended, the LuSens test may be performed.
ACCEPTABILITY CRITERIA
The following acceptance criteria should be met in the LuSens test method (OECD 442D):
• The average luciferase activity induction obtained with the positive control, 120 μM EGDMA should be ≥ 2.5, and the positive control should have a relative cell viability ≥ 70% as compared to the solvent control.
• The average luciferase activity induction obtained with the negative control, 5000 μM Lactic acid, as well as the basal expression of untreated cells should be < 1.5 fold as compared to the average solvent control.
• The average coefficient of variation (CV%) of the luminescence reading for the solvent controls (DMSO) should be below 20% in each main experiment.
• At least three test item concentrations should have cell viability of at least 70% relative to the solvent controls. Moreover, in case a result is to be considered negative, at least one concentration should be cytotoxic, i.e. have a cell viability < 70%, or the maximum concentration of 2000 μM (or 2000 μg/ mL for substances with no defined MW) should have been tested. - Vehicle / solvent control:
- DMSO
- Negative control:
- DL-Lactic acid
- Positive control:
- other: EGDMA (120 µM) [442D]
Results and discussion
- Positive control results:
- The average luciferase activity induction obtained with the positive control, 120 μM EGDMA was ≥ 2.5 (Experiment 1: 5.812; Experiment 2: 5.817).
The positive control had a relative cell viability ≥ 70% as compared to the solvent control (Experiment 1: 101.842; Experiment 2: 80.545).
In vitro / in chemico
Resultsopen allclose all
- Group:
- test chemical
- Run / experiment:
- run/experiment 1
- Parameter:
- EC 1.5 [442D]
- Cell viability:
- Cell viability was < 70% at 665 and 798 µM
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: luciferase induction was < 1.5 fold compared to the solvent control at all the tested concentrations
- Group:
- test chemical
- Run / experiment:
- run/experiment 2
- Parameter:
- EC 1.5 [442D]
- Cell viability:
- Cell viability was > 70% at all tested concentrations
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: luciferase induction was < 1.5 fold compared to the solvent control at all the tested concentrations
- Outcome of the prediction model:
- negative [in vitro/in chemico]
- Other effects / acceptance of results:
- CYTOTOXICITY DOSE RANGE FINDER:
- In the cytotoxicity test, cytotoxic effects were observed following incubation with the test item starting with the concentration of 1000 µM up to the highest tested concentration of 2000 µM (threshold of cytotoxicity: < 75%). The CV75 value of the cytotoxicity test was calculated as 665.1 µM.
MAIN EXPERIMENT (LuSens and MTT):
- MTT cytotoxicity assay: in Experiment 1, cytotoxicity (cell viability < 70%) was observed at 665 and 798 µM. In Experiment 2, no cytotoxicity was observed (cell viability was >70% at all tested concentrations).
- LuSens assay: after treatment with the test item for 48 ± 1 hours the luciferase induction was < 1.5 fold compared to the solvent control in at least 2 consecutive tested concentrations. Since these conditions are met in 2 out of 2 main experiments, the LuSens prediction was considered negative (tabulated results are presented in section "any other information on results incl. tables").
ACCEPTANCE OF RESULTS:
The acceptance criteria were met:
- The average luciferase activity induction obtained with the positive control, 120 μM EGDMA was ≥ 2.5 (ME 1: 5.812; ME 2: 5.817).
- The positive control had a relative cell viability ≥ 70% as compared to the solvent control (ME 1: 101.842; ME 2: 80.545).
- The average luciferase activity induction obtained with the negative control, 5000 μM Lactic acid, as well as the basal expression of untreated cells was < 1.5 fold as compared to the average solvent control (ME 1: 1.023; ME 2: 0.967).
- At least three test concentrations had a cell viability of at least 70% relative to the solvent/vehicle controls. Moreover, since the result is considered negative, at least one concentration was cytotoxic, i.e. had a cell viability < 70%, or the maximum concentration of 2000 μM have been tested.
- Historical control data is included in the attached background material.
Any other information on results incl. tables
Table 1: Results of the Dose Finding Assay (Cytotoxicity Test)
Treatment Group |
Concentration |
OD570 |
Mean OD570 |
SD OD570 |
Mean OD570 blank corr. |
Cell viability [%] |
||||
Well 1 |
Well 2 |
Well 3 |
||||||||
Blank |
- |
0.015 |
- |
- |
- |
- |
- |
- |
||
Solvent control |
- |
- |
- |
- |
0.598 |
0.035 |
0.583 |
100.00 |
||
Medium control |
- |
- |
- |
- |
0.811 |
0.081 |
0.796 |
136.67 |
||
Positive Control |
|
120 |
µM |
0.608 |
0.659 |
- |
0.634 |
0.036 |
0.619 |
106.15 |
Negative Control |
|
5000 |
µM |
0.630 |
0.647 |
0.574 |
0.617 |
0.038 |
0.602 |
103.32 |
Test Item |
C1 |
0.98 |
µM |
0.527 |
0.628 |
0.547 |
0.567 |
0.053 |
0.552 |
94.79 |
C2 |
1.95 |
µM |
0.602 |
0.502 |
0.498 |
0.534 |
0.059 |
0.519 |
89.07 |
|
C3 |
3.91 |
µM |
0.600 |
0.589 |
0.598 |
0.596 |
0.006 |
0.581 |
99.66 |
|
C4 |
7.81 |
µM |
0.498 |
0.523 |
0.503 |
0.508 |
0.013 |
0.493 |
84.61 |
|
C5 |
15.6 |
µM |
0.536 |
0.580 |
0.476 |
0.531 |
0.052 |
0.516 |
88.50 |
|
C6 |
31.3 |
µM |
0.491 |
0.556 |
0.532 |
0.526 |
0.033 |
0.511 |
87.76 |
|
C7 |
62.5 |
µM |
0.562 |
0.556 |
0.541 |
0.553 |
0.011 |
0.538 |
92.33 |
|
C8 |
125 |
µM |
0.529 |
0.622 |
0.604 |
0.585 |
0.049 |
0.570 |
97.83 |
|
C9 |
250 |
µM |
0.516 |
0.488 |
0.504 |
0.503 |
0.014 |
0.488 |
83.70 |
|
C10 |
500 |
µM |
0.483 |
0.460 |
0.448 |
0.464 |
0.018 |
0.449 |
77.00 |
|
C11 |
1000 |
µM |
0.441 |
0.451 |
0.393 |
0.428 |
0.031 |
0.413 |
70.94 |
|
C12 |
2000 |
µM |
0.337 |
0.297 |
0.335 |
0.323 |
0.023 |
0.308 |
52.86 |
The CV75 value of the cytotoxicity test was calculated as 665.1 µM.
Table 2: Results of the main experiment 1 (Cell viability)
Treatment Group |
Concentration |
OD570 |
Mean OD570 |
SD OD570 |
Mean OD570 blank corr. |
Cell viability [%] |
|||||||
Well 1 |
Well 2 |
Well 3 |
Well 4 |
Well 5 |
Well 6 |
||||||||
Blank |
- |
0.014 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
||
Solvent control |
- |
- |
- |
- |
- |
- |
- |
0.350 |
0.056 |
0.336 |
100.00 |
||
Medium control |
- |
- |
- |
- |
- |
- |
- |
0.337 |
0.089 |
0.323 |
96.15 |
||
Positive Control |
|
120 |
µM |
0.282 |
0.372 |
0.430 |
0.298 |
0.400 |
- |
0.356 |
0.064 |
0.342 |
101.84 |
Negative Control |
|
5000 |
µM |
0.457 |
0.373 |
0.393 |
0.362 |
0.269 |
0.369 |
0.371 |
0.061 |
0.357 |
106.04 |
Test Item |
C1 |
321 |
µM |
0.430 |
0.428 |
0.440 |
- |
- |
- |
0.433 |
0.006 |
0.419 |
124.53 |
C2 |
385 |
µM |
0.399 |
0.370 |
0.377 |
- |
- |
- |
0.382 |
0.015 |
0.368 |
109.46 |
|
C3 |
462 |
µM |
0.426 |
0.385 |
0.378 |
- |
- |
- |
0.396 |
0.026 |
0.382 |
113.72 |
|
C4 |
554 |
µM |
0.311 |
0.349 |
0.360 |
- |
- |
- |
0.340 |
0.026 |
0.326 |
96.96 |
|
C5 |
665 |
µM |
0.172 |
0.177 |
0.183 |
- |
- |
- |
0.177 |
0.006 |
0.163 |
48.58 |
|
C6 |
798 |
µM |
0.130 |
0.197 |
0.175 |
- |
- |
- |
0.167 |
0.034 |
0.153 |
45.61 |
Table 3: Results of the main experiment 1 (Fold induction)
Treatment Group |
Concentration |
Luminescence |
Mean Luminescence |
SD Luminescence |
Mean Luminescence blank corr. |
Fold Induction |
|||||||
Well 1 |
Well 2 |
Well 3 |
Well 4 |
Well 5 |
Well 6 |
||||||||
Blank |
- |
177 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
||
Solvent control |
- |
- |
- |
- |
- |
- |
- |
292.792 |
22.853 |
115.792 |
1.00 |
||
Medium control |
- |
- |
- |
- |
- |
- |
- |
324.583 |
18.676 |
147.583 |
1.275 |
||
Positive Control |
|
120 |
µM |
835 |
776 |
732 |
939 |
968 |
|
850.000 |
101.821 |
673.000 |
5.812 |
Negative Control |
|
5000 |
µM |
288 |
333 |
266 |
288 |
273 |
325 |
295.500 |
27.443 |
118.500 |
1.023 |
Test Item |
C1 |
321 |
µM |
303 |
296 |
310 |
- |
- |
- |
303.000 |
7.000 |
126.000 |
1.088 |
C2 |
385 |
µM |
281 |
266 |
296 |
- |
- |
- |
281.000 |
15.000 |
104.000 |
0.898 |
|
C3 |
462 |
µM |
296 |
273 |
318 |
- |
- |
- |
295.667 |
22.502 |
118.667 |
1.025 |
|
C4 |
554 |
µM |
310 |
333 |
325 |
- |
- |
- |
322.667 |
11.676 |
145.667 |
1.258 |
|
C5 |
665 |
µM |
266 |
273 |
355 |
- |
- |
- |
298.000 |
49.487 |
121.000 |
1.045 |
|
C6 |
798 |
µM |
273 |
303 |
288 |
- |
- |
- |
288.000 |
15.000 |
111.000 |
0.959 |
Table 4: Results of the main experiment 2 (Cell viability)
Treatment Group |
Concentration |
OD570 |
Mean OD570 |
SD OD570 |
Mean OD570 blank corr. |
Cell viability [%] |
|||||||
Well 1 |
Well 2 |
Well 3 |
Well 4 |
Well 5 |
Well 6 |
||||||||
Blank |
- |
0.014 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
||
Solvent control |
- |
- |
- |
- |
- |
- |
- |
0.534 |
0.128 |
0.520 |
100.00 |
||
Medium control |
- |
- |
- |
- |
- |
- |
- |
0.605 |
0.115 |
0.591 |
113.70 |
||
Positive Control |
|
120 |
µM |
0.486 |
0.528 |
0.413 |
0.390 |
0.346 |
- |
0.433 |
0.074 |
0.419 |
80.55 |
Negative Control |
|
5000 |
µM |
0.541 |
0.611 |
0.576 |
0.554 |
0.495 |
0.496 |
0.546 |
0.045 |
0.532 |
102.27 |
Test Item |
C1 |
321 |
µM |
0.496 |
0.524 |
0.463 |
- |
- |
- |
0.494 |
0.031 |
0.480 |
92.42 |
C2 |
385 |
µM |
0.504 |
0.501 |
0.476 |
- |
- |
- |
0.494 |
0.015 |
0.480 |
92.30 |
|
C3 |
462 |
µM |
0.500 |
0.594 |
0.558 |
- |
- |
- |
0.551 |
0.047 |
0.537 |
103.26 |
|
C4 |
554 |
µM |
0.479 |
0.484 |
0.489 |
- |
- |
- |
0.484 |
0.005 |
0.470 |
90.44 |
|
C5 |
665 |
µM |
0.367 |
0.484 |
0.438 |
- |
- |
- |
0.430 |
0.059 |
0.416 |
79.98 |
|
C6 |
798 |
µM |
0.395 |
0.471 |
0.446 |
- |
- |
- |
0.437 |
0.039 |
0.423 |
81.46 |
Table 5: Results of the main experiment 2 (Fold induction)
Treatment Group |
Concentration |
Luminescence |
Mean Luminescence |
SD Luminescence |
Mean Luminescence blank corr. |
Fold Induction |
|||||||
Well 1 |
Well 2 |
Well 3 |
Well 4 |
Well 5 |
Well 6 |
||||||||
Blank |
- |
155 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
||
Solvent control |
- |
- |
- |
- |
- |
- |
- |
263.542 |
25.044 |
108.542 |
1.00 |
||
Medium control |
- |
- |
- |
- |
- |
- |
- |
278.500 |
25.508 |
123.500 |
1.138 |
||
Positive Control |
|
120 |
µM |
732 |
909 |
761 |
791 |
739 |
- |
786.400 |
72.290 |
631.400 |
5.817 |
Negative Control |
|
5000 |
µM |
229 |
259 |
266 |
244 |
281 |
281 |
260.000 |
20.669 |
105.000 |
0.967 |
Test Item |
C1 |
321 |
µM |
347 |
244 |
273 |
- |
- |
- |
288.000 |
53.113 |
133.000 |
1.225 |
C2 |
385 |
µM |
303 |
288 |
222 |
- |
- |
- |
271.000 |
43.093 |
116.000 |
1.069 |
|
C3 |
462 |
µM |
303 |
296 |
266 |
- |
- |
- |
288.333 |
19.655 |
133.333 |
1.228 |
|
C4 |
554 |
µM |
251 |
236 |
236 |
- |
- |
- |
241.000 |
8.660 |
86.000 |
0.792 |
|
C5 |
665 |
µM |
244 |
251 |
251 |
- |
- |
- |
248.667 |
4.041 |
93.667 |
0.863 |
|
C6 |
798 |
µM |
266 |
266 |
273 |
- |
- |
- |
268.333 |
4.041 |
113.333 |
1.044 |
Applicant's summary and conclusion
- Interpretation of results:
- other: no skin sensitising potential based on the key event “inflammatory response in keratinocytes”
- Conclusions:
- The study was performed according to OECD guideline 442D. The test item did not activate the LuSens cells up to a concentration of 798 µg/mL under the test conditions of this study. Therefore, the test item is considered negative for the second key event of the skin sensitisation Adverse Outcome Pathway (AOP).
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