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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 268-040-3 | CAS number: 67990-17-4
- 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 chemico
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- 16 January 2019 - 21 March 2019
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 018
- Report date:
- 2019
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 442C (In Chemico Skin Sensitisation: Direct Peptide Reactivity Assay (DPRA))
- Version / remarks:
- 2015
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of study:
- direct peptide reactivity assay (DPRA)
Test material
- Reference substance name:
- Sodium (2-butoxyethoxy)acetate
- EC Number:
- 268-040-3
- EC Name:
- Sodium (2-butoxyethoxy)acetate
- Cas Number:
- 67990-17-4
- Molecular formula:
- C8H16O4.Na
- IUPAC Name:
- sodium (2-butoxyethoxy)acetate
- Test material form:
- liquid: viscous
- Details on test material:
- Batch Number: P59564
Appearance: white-yellow, low melting point paste
Purity: 100 %
Expiration date: 31.12.2018
Storage: room temperature (ca. 20°C)
Constituent 1
In chemico test system
- Details on the study design:
- Synthetic peptides:
Sequence Cys-Peptide (Cysteine): Ac-RFAACAA-COOH (MW = 750.9 g/mol), Batch No.:150219HS-MHe
Sequence Lys-Peptide (Lysine): Ac-RFAAKAA-COOH (MW = 775.9 g/mol), Batch No.: 020517HS_MHEW0119-2
Instruments and Devices:
Analytical and precision scales
Volumetric measurement tools
pH-meter
Standard laboratory glassware
Incubation chamber
Chemicals:
Water for chromatography
H2O, HPLC grade
Demineralised water
Acetonitrile for chromatography
CH3CN, ACN, HPLC grade
Trifluoroacetic acid
TFA, for spectroscopy, Merck
Isopropanol
CH3CHOHCH3, p.a.
Buffers:
Phosphate buffer pH 7.5 ± 0.05
18 (v/v) % 0.1 M sodium phosphate monobasic (of Sodium Phosphate Monobasic Monohydrate (NaH2PO4 • H2O) in purified water)
82 (v/v) % 0.1 M sodium phosphate dibasic (of Sodium Phosphate Dibasic Heptahydrate (Na2HPO4 • 7H2O) in purified water) pH was adjusted with either the monobasic or dibasic solution
Ammonium acetate buffer pH 10.2 ± 0.05
0.1 M Ammonium Acetate (CH3CO2NH4) in purified water
pH was adjusted with Ammonium Hydroxide (NH4OH) drop by drop
Positive control:
Cinnamaldehyde (CAS 104-55-2, 99.5 %,100 mM solution in acetonitrile for the cysteine peptide
100 mM solutions of the positive control chemical in acetonitrile were prepared just before use.
Peptide stock solutions
Cysteine peptide stock solution, 0.667 mM, 0.501 mg/mL:
The needed amount of peptide was calculated based on the equation below (0.01085 g ± 10%). The previously calculated amount of the peptide (0.01153 g) was pre-weighted and 21.25 mL of pH 7.5 phosphate buffer was added right before beginning the assay in the valid run.
Lysine peptide stock solution, 0.667 mM, 0.518 mg/mL:
The needed amount of peptide was calculated based on the equation below (0.01065 g ± 10%). The previously calculated amount of the peptide (0.01003 g) was pre-weighted and 18.83 mL of pH 10.2 acetate buffer was added right before beginning of the assay in the valid run.
Test item stock solution:
100 mM solutions of the test chemical in the appropriate solvent were prepared just before use. The needed amount of test chemical was calculated (0.0991 g ± 10 %) and weighted based on the single average molecular weight. 0.0993 g test chemical was weighted for the stock solution used for the cysteine peptide depletion determination and 0.0991 g test chemical was weighted for the stock solution used for lysine peptide depletion determination in the valid runs.
Test item samples:
Samples are prepared in triplicate for each peptide. The Cys-peptide samples are prepared in 1:10 molar ratio (0.5 mM peptide: 5 mM test item), the Lys-peptide samples in 1:50 molar ratio (0.5 mM peptide and 25 mM test item) using the stock solutions described. A final volume of 1 mL per sample is prepared for each sample
Incubation:
The positive control, solvent control and test item samples are incubated in closed amber glass HPLC vials in an incubation chamber at 25.0 ± 2.5 °C for 24 ± 2 h. When the test item stock solution was combined with the cysteine and lysine stock solutions (reaction samples) or the phosphate and ammonium acetate buffer (co-elution controls), the samples appeared to be clear and homogenous.
Measurements
HPLC system with UV/VIS-Detector
Evaluation of results:
Evaluation criteria of results according to the cysteine 1:10 / lysine 1:50 prediction model.
Mean peptide depletion [%] Reactivity Evaluation
> 42.47 high reactivity positive
> 22.62 ≤ 42.47 moderate reactivity positive
> 6.38 ≤ 22.62 low reactivity positive
0- ≤ 6.385 minimal or no reactivity negative
Acceptance criteria
-the standard calibration curve should have an r2 > 0.99
-the mean peptide concentration of reference controls A should be 0.50 ± 0.05 mM and the coefficient of variation (CV) of peptide peak areas for the nine reference controls B and C in acetonitrile should be ≤ 15.0%.
-the mean percent peptide depletion value of the three replicates for the positive control cinnamaldehyde should be between 60.8% and 100% for the cysteine peptide and between 40.2% and 69.0% for the lysine peptide and the maximum standard deviation (SD) for the positive control replicates should be < 14.9% for the percent cysteine depletion and < 11.6% for the percent lysine depletion
The following criteria should be met for a test chemical’s results to be considered valid:
-the maximum standard deviation for the test chemical replicates should be less than 14.9 % for the percent cysteine depletion and less than 11.6% for the percent lysine depletion
-the mean peptide concentration of the three reference controls C in the appropriate solvent should be 0.50 ± 0.05 mM.
Results and discussion
- Positive control results:
- The acceptance criteria were met for the positive control with a cysteine peptide depletion value of 70.60 % and a mean lysine peptide depletion value of 53.00 %. The standard deviation of the percent peptide depletions of the positive control was 1.05 % and 0.49 % for the cysteine and lysine depletion, respectively. Thus, this validity criterion was also met.
In vitro / in chemico
Results
- Key result
- Run / experiment:
- other: 1st experiment
- Parameter:
- other: mean peptide depletion of both peptides (%)
- Value:
- 2.44
- Vehicle controls validity:
- valid
- Negative controls validity:
- not examined
- Positive controls validity:
- valid
- Other effects / acceptance of results:
- OTHER EFFECTS:
Co-elution:
The test chemical did not absorb at 220 nm significantly (<10 % compared to the respective reference control) when tested with the cysteine and lysine peptides. The range of retention time for cysteine peptide was between 8.383 and 8.480 and the range of retention time for lysine peptide was between 6.118 and 6.317.
DEMONSTRATION OF TECHNICAL PROFICIENCY: Prior to routine use of the method, TOXI-COOP ZRT. demonstrated technical proficiency in a separate study by correctly obtaining the expected DPRA prediction for 10 proficiency substances as recommended in the OECD TG 442C guideline.
ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
- Acceptance criteria met for variability between replicate measurements: yes
Any other information on results incl. tables
Table 1: Cysteine peptide depletion values for the positive control and the test item
Sample |
Peptide conc. calculated |
Mean peptide conc. |
Peptide depletion |
Standard deviation (SD) for peptide depletion (%) |
ref C upw, rep I |
0.50 |
0.49 |
- |
- |
ref C upw, rep II |
0.49 |
- |
||
ref C upw, rep III |
0.49 |
- |
||
CINNAMALDEHYDE, rep I |
0.14 |
0.14 |
71.81% |
1.05% |
CINNAMALDEHYDE, rep II |
0.15 |
70.09% |
||
CINNAMALDEHYDE, rep III |
0.15 |
69.91% |
||
sodium (2-butoxyethoxy) acetate, rep I |
0.48 |
0.47 |
4.41% |
1.72% |
sodium (2-butoxyethoxy) acetate, rep II |
0.48 |
1.57% |
||
sodium (2-butoxyethoxy) acetate, rep III |
0.47 |
4.69% |
(upw = ultrapure water)
Table 2: Lysine peptide depletion values for the positive control and the test item
Sample |
Peptide conc. calculated |
Mean peptide conc. |
Peptide depletion |
Standard deviation (SD) for peptide depletion (%) |
ref C upw, rep I |
0.49 |
0.49 |
- |
- |
ref C upw, rep II |
0.49 |
- |
||
ref C upw, rep III |
0.49 |
- |
||
CINNAMALDEHYDE, rep I |
0.23 |
0.23 |
53.35% |
0.49% |
CINNAMALDEHYDE, rep II |
0.24 |
52.44% |
||
CINNAMALDEHYDE, rep III |
0.23 |
53.22% |
||
sodium (2-butoxyethoxy) acetate, rep I |
0.49 |
0.49 |
1.17% |
0.19% |
sodium (2-butoxyethoxy) acetate, rep II |
0.49 |
1.53% |
||
sodium (2-butoxyethoxy) acetate, rep III |
0.49 |
1.27% |
Table 3: Mean peptide depletion values for the positive control and the test chemical
Name, replicate number |
Obtained mean % cysteine peptide depletion |
Obtained mean % lysine peptide depletion |
Mean % obtained peptide depletion |
sodium (2-butoxyethoxy) acetate |
3.56 |
1.32 |
2.44 |
CINNAMALDEHYDE |
70.60 |
53.00 |
61.80 |
Applicant's summary and conclusion
- Interpretation of results:
- other: no peptide depletion
- Remarks:
- 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.
- Conclusions:
- Based on these results and the cysteine 1:10 / lysine 1:50 prediction model, the test item was concluded to have no or minimal reactivity towards the synthetic peptides and thus is not a potential skin sensitizer under the experimental conditions of the in chemico Direct Peptide Reactivity Assay (DPRA) method.
- Executive summary:
A study according OECD TG 442 C was conducted for detection of the sensitising potential of the test item by quantifying its reactivity towards synthetic peptides containing either lysine or cysteine.
For the test chemical and positive control substance, in order to derive a prediction five independent tests were conducted, since the first run with cysteine peptide and the first two runs with lysine peptide did not meet acceptance criteria those results were rejected. All in all, the results of the two valid runs were used for the classification of the test item.
Peptide depletion resulted from the positive control cinnamaldehyde was 70.60 % with cysteine peptide and 53.00 % with the lysine peptide while the standard deviation (SD) of the percent peptide depletions were 1.05 % and 0.49 % for the cysteine and lysine peptides respectively in the valid runs. The back-calculated values of the reference control replicates were within the expected molarity concentration range for the cysteine (0.50 – 0.49 mM) and lysine peptides (0.50 – 0.49 mM) and the CV % for the nine reference controls B and C in acetonitrile were 2.6 % and 0.3 % percentages for the cysteine and lysine peptides. In the last runs for each peptide all validity criteria were also met, confirming the validity of the assay.
The percent cysteine peptide depletion value of the test item was 3.56 % while the percent lysine peptide depletion was 1.32 %. The mean depletion value of the peptides was used to categorize the test chemical in one of the four classes of reactivity. No co-elution was observed with either cysteine or lysine peptides; therefore the cysteine 1:10 / lysine 1:50 prediction model was used for the discrimination between sensitisers and non-sensitisers. The mean peptide depletion of the test item was 2.44 %, which did not exceed the 6.38 % threshold of the applicable prediction model.
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.