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EC number: 603-333-5 | CAS number: 129423-55-8
- 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
Endpoint summary
Administrative data
Description of key information
Based upon results Dioctyl dodecyl dodecanedioate is predicted as a non sensitizer.
Key value for chemical safety assessment
Skin sensitisation
Link to relevant study records
- Endpoint:
- skin sensitisation: in vitro
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- From 11/03/2019 to 05/04/2019
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 442E (In Vitro Skin Sensitisation assays addressing the key event on activation of dendritic cells on the Adverse Outcome Pathway for skin sensitisation)
- Principles of method if other than guideline:
- The Test item was dissolved in non-standard vehicle Acetone (0.1 %in-well concentration)
- GLP compliance:
- not specified
- Remarks:
- Under a GLP-like quality system performed, where instruments and equipment are calibrated and maintained according to the supplier, a strict protocol is followed, QC criteria are met and notes and raw data is archived in paper or electronic.
- Type of study:
- other: GARDskin
- Details of test system:
- other: GARDskin
- Details on the study design:
- STANDARD GARD ASSAY PROTOCOL
Cell line maintenance and seeding of cells for stimulation
The human myeloid leukemia-derived cell line SenzaCell (available through ATCC), acting as an in vitro model of human Dendritic Cell (DC), is maintained in a-MEM (Thermo Scientific Hyclone, Logan, UT) supplemented with 20% (volume/volume) fetal calf serum (Life Technologies, Carlsbad, CA) and 40 ng/ml recombinant human Granulocyte Macrophage Colony Stimulating Factor (rhGM-CSF) (Miltenyi Biotec, Germany). A media change during expansion is performed every 3-4 days. Working stocks of cultures are grown for a maximum of 16 passages or two months after thawing. For chemical stimulation of cells, exposed cells are incubated for 24 hat 37°C, 5% CO2 and 95% humidity.
Test items handling and assessment of cytotoxicity
All Test items were stored according to instructions from the Sponsor, to ensure stability of Test items. Test items were dissolved in DMSO or water, based on physical properties. As many Test Items will have a toxic effect on the cells, cytotoxic effects of Test Items were monitored. When Test Items were poorly dissolved in cell medium the maximum soluble concentration was assessed. The assayed Test Items were titrated to concentrations ranging from 1 µM to the maximum soluble concentration in cell media. For freely soluble Test Iterns, 500 µM was set as the upper limit of the titration range. For Test Items dissolved in DMSO, the in-well concentration of DMSO was 0.1%. After incubation for 24 hat 37°C, 5% CO2 and 95% humidity, harvested cells were stained with the viability marker Propidium Iodide (PI) (BD Bioscience, USA) and analyzed by flow cytometry. PI-negative cells were defined as viable, and the relative viability of cells stimulated with each concentration in the titration range was calculated as
Relative viability = fraction of viable stimulated cells / fraction of viable unstimulated cells * 100
For toxic Test Items, the concentration yielding 90% relative viability (Rv90) was used for the GARD Assay, the reason being that this concentration demonstrates bioavailability of the test substance used for stimulation, while not impairing immunological responses. For non-toxic Test Items, a concentration of 500 µM was used if possible. For non-toxic Test Items that were insoluble at 500 µM in cell media, the highest soluble concentration was used. Whichever of these three criteria was met, only one concentration will be used for gene expression analysis. The concentration to be used for any given chemical was termed the 'GARD input concentration'.
GARD Main Stimulation
Once the GARD input concentration for the Test Items was established, the cells were stimulated again, as described above, using the GARD input concentration only. In addition to the assayed Test Items a set of positive and negativecontrols were performed as reference and quality controls. The Test Items and controls were assayed in biological replicates (n 2), performed at different time-points and using different cell cultures. After incubation for 24 hat 37°C, 5% CO2and 95% humidity, cell culture was lysed in TRizol reagent (Life Technologies) and stored at -20°C until RNA was extracted. In parallel, stimulated cells were PI stained and analysed by flow cytometry to verify the expected relative viability.
Isolation of RNA
RNA isolation from lysed cells was performed using commercially available kits (Direct-Zol RNA MiniPrep, Zymo Research, Irvine, CA). Total RNA was quantified and quality controlled using BioAnalyzer equipment (Agilent, Santa Clara, CA).
GARDskin Endpoint Measurement
Gene expression analysis using Nanostring nCounter System
A total of 100 ng of RNA was used as sample input in a hybridization assay with the GPS (GARD Prediction Signature) specific Reporter CodeSet (Nanostring Technologies, Seattle, WA). The hybridized sample was prepared on chip using nCounter Prep Station and individual transcripts of the GPS were quantified using Nanostring Digital Analyzer (Nanostring Technologies, Seattle, WA).
Nanostring nCounter data acquisition and normalization
Raw data was exported from the Digital Analyzer and RCC-files were imported into the R environment for statistical computing (www.r-project.org). Raw data was normalized using a single-chip normalization algorithm based on gene specific Counts Per Total Counts (CPTC) algorithm.
Prediction model and data analysis
For assessment of skin sensitization, a Support Vector Machine (SVM) was modelled on a training data set corresponding to samples used for assay development. For a comprehensive overview of the training data set and methods, see Forreryd et al, 2016 (for further details on biomarker discovery, see Johansson et al., 2011). Batch variations between the training data set and the test data set were eliminated using the Batch Adjustment by Reference Alignment (BARA) method (Gradin et al., 2019), using unstimulated cells as a reference control. Each sample in the test set were assigned a Decision Value (DV), based on its transcriptional levels of the GPS biomarker signature. Any test substance with a mean DV o (n 2) isclassified as a skin sensitizer.
The Test item was stored and handled according to instructions provided by the Sponsor.
For Dioctyl dodecyl dodecanedioate deviations from standard GARD assay protocol was made, where the Test item was dissolved in non-standard vehicle Acetone (0.1 %in-well concentration).
TheTest item was assessed for solubility and cytotoxic effect in order to establish the GARD input concentration (concentration inducing 90% relative viability). The Test item showed neither solubility issues nor cell cytotoxicity, therefore the GARD input concentration was determined to 500 µM.
For the GARD prediction, all replicates of the Test items were assigned Decision Values, where a Positive and a Negative Decision Value predicts the Test item as a sensitizer and a non-sensitizer, respectively. To eliminate cell batch variations between the training data set and the test data, unstimulated cells (m:2) were included as reference controls as described in the Standard GARD Assay Protocol section in this report. TheGARD assay passes Quality Control when the positive and the negative controls are predicted as a Sensitizer (+) and a Non-sensitizer(-), respectively. - Vehicle / solvent control:
- other: Acetone
- Negative control:
- other: DMSO
- Positive control:
- other: p-phenulendiamine (PPD)
- Positive control results:
- Positive control of p-phenulendiamine (PPD) showed decision value (mean ±SD) of 6.59 ± 0.39
- Key result
- Group:
- test chemical
- Run / experiment:
- run/experiment 1
- Parameter:
- other: Decision Value
- Value:
- -1.01
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- no indication of skin sensitisation
- Other effects / acceptance of results:
- The objective of this study was to evaluate Dioctyl dodecyl dodecanedioate provided by the Sponsor using the GARDskin assay for a binary prediction as a sensitizer or a non sensitizer.
Based upon results of this study, Dioctyl dodecyl dodecanedioate is predicted as a non sensitizer. However, since no cytotoxic effect is seen, the bioavailability of the Test item is uncertain. Therefore, the prediction of Dioctyl dodecyl dodecanedioate as a non-sensitizer should be interpreted with caution. - Deviation from standard GARD assay protocol using non-standard vehicle
- The highest soluble concentration used in screening
- Concentration of Test item inducing 90% Relative Viability
- Concentration based on screen and Rv90
- Interpretation of results:
- GHS criteria not met
- Conclusions:
- The objective of this study was to evaluate Dioctyl dodecyl dodecanedioate provided by the Sponsor using the GARDskin assay for a binary prediction as a sensitizer or a non sensitizer.
Based upon results of this study, Dioctyl dodecyl dodecanedioate is predicted as a non sensitizer. - Executive summary:
The objective of this study was to evaluate Dioctyl dodecyl dodecanedioate provided by the Sponsor to assess the sensitizing hazard using the GARDskin assay for binary prediction as a sensitizer or a non-sensitizer.
Based upon the results of this study, Dioctyl dodecyl dodecanedioate is predicted as a non sensitizer.
Reference
Results from the cell cytotoxicity screen and the obtained GARD input concentrations for the provided Test item is presented in Table 1.
Table 1. Test item details
Test item |
Vehicle• | Max. screenII (µM) | Rv9om (µM) | GARD input concIV (µM) |
Dioctyl dodecyl dodeca.n. edioate | Acetone | 500 | No cytotox | 500 |
The non-standard vehicle (Acetone) and the Test item were predicted as GARDskin non-sensitizers.
Test item | Decision Value (Mean:tSD) | GARDskin Prediction | GARDskin assay Quality Control |
Dioctyl dodecyl dodecanedioate | -1.01 ±0.72 | Non-sensitizer | PASSED |
Acetone | -0.78 ±0.82 | Non-sensitizer | PASSED |
Positive ctrl, PPD | 6.59 ±0.39 | Sensitizer | PASSED |
Negative ctrl, DMSO | -1.51 ±0.30 | Non-sensitizer | PASSED |
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (not sensitising)
- Additional information:
No cytotoxic effects were seen
Justification for classification or non-classification
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
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