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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 950-718-1 | 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
Skin sensitisation
Administrative data
- Endpoint:
- skin sensitisation: in chemico
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- The study was conducted between 08 January 2019 and 11 January 2019.
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 019
- 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))
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of study:
- direct peptide reactivity assay (DPRA)
- Justification for non-LLNA method:
- The DPRA test allows quantification of a chemical’s reactivity and is used to categorize a substance in one of four classes of reactivity to allow discriminating between skin sensitizing and non-sensitizing chemicals and thus assesses their sensitization potential.
Test material
- Reference substance name:
- Reaction mass of (E)-1-(3,6-dimethylcyclohex-3-en-1-yl)-2-methylpent-1-en-3-one and (E)-1-(4,6-dimethylcyclohex-3-en-1-yl)-2-methylpent-1-en-3-one
- EC Number:
- 950-718-1
- Molecular formula:
- C14H22O
- IUPAC Name:
- Reaction mass of (E)-1-(3,6-dimethylcyclohex-3-en-1-yl)-2-methylpent-1-en-3-one and (E)-1-(4,6-dimethylcyclohex-3-en-1-yl)-2-methylpent-1-en-3-one
- Test material form:
- liquid
Constituent 1
- Specific details on test material used for the study:
- Identification PG-RAW-90-032
Appearance: Clear liquid
Storage conditions: Refrigerated temperature (2°C to 8°C)
In chemico test system
- Details on the study design:
- Peptide and Positive Control
Synthetic peptide containing Cysteine
Alternative name: Ac-RFAACAA-OH
Batch number: 1857724
Stated purity: 96.2% (by HPLC)
Molecular Weight: 751 g/mol
Supplier: AnaSpec
Storage conditions: Frozen (-10°C to -30°C)
Synthetic peptide containing Lysine
Alternative name: Ac-RFAAKAA-OH
Batch number: 1658141
Stated purity: 96.8% (by HPLC).
Molecular Weight: 776 g/mol
Supplier: AnaSpec
Storage conditions: Frozen (-10°C to -30°C)
Cinnamic Aldehyde (Positive control)
Batch number: MKCB9907
Stated purity: 99.1%
Molecular Weight: 132 g/mol
Supplier: SAFC
Storage conditions: Room temperature (15°C to 25°C)
Expiry/retest date: November 2021
Apparatus
High performance liquid chromatograph (HPLC): Waters Alliance 2695 separation module and 2487 dual wavelength detector
Balances fitted with printers: Capability of weighing to 5 decimal places
General laboratory apparatus and glassware.
Analytical Procedure
Reagents
Acetonitrile (ACN): HPLC gradient grade
Trifluoroacetic acid (TFA): 99% Pure
Water: Deionised reverse osmosis
Ammonium Acetate: Analytical reagent
Sodium Phosphate, monobasic: Analytical reagent
Sodium Phosphate, dibasic: Analytical reagent
Ammonium Hydroxide: Analytical reagent
100 mM Phosphate buffer, pH 7.5: In house preparation
100 mM Ammonium Acetate buffer, pH 10.2: In house preparation
HPLC Mobile Phase A: 0.1% v/v TFA in Water, in house preparation
HPLC Mobile Phase B: 0.085% v/v TFA in ACN, in house preparation
Assessment of Test Item Solubility
The solubility of PG-RAW-90-032 was assessed in acetonitrile at a nominal concentration of 100 mM.
Preparation of Peptide Stock Solutions
Stock solutions of each peptide at concentrations of 0.667 mM were prepared by dissolution of pre-weighed aliquots of the appropriate peptide in ca 20 mL aliquots of the appropriate buffer solution (Cysteine in 100 mM phosphate buffer pH 7.5, Lysine in 100 mM Ammonium acetate buffer pH 10.2).
Preparation of Peptide Calibration Standards
Calibration standards of both peptides were prepared by diluting the requisite stock solution in the appropriate buffer and acetonitrile and contained each peptide at concentrations of 0.0167 mM, 0.0334 mM, 0.0667 mM, 0.133 mM, 0.267 mM and 0.534 mM. A buffer blank was also prepared.
Preparation of Stability Controls and Precision Control
Stability controls (Reference Control B), precision controls of both peptides were prepared at a concentration of 0.5 mM in acetonitrile/buffer.
Preparation of Positive Control Solution and Test Item Stock Solution
The positive control chemical (Cinnamic Aldehyde) was prepared at a concentration of 100 mM in acetonitrile. A 100 mM stock solution of PG-RAW-90-032 was also prepared in acetonitrile.
Preparation of Positive Control and Cysteine Peptide Depletion Samples and Co-elution Controls
Triplicate solutions each of the positive control and PG-RAW-90-032 stock solutions were diluted with the Cysteine peptide stock solution so as to prepare solutions containing 0.5 mM Cysteine and 5 mM of Cinnamic Aldehyde or 5 mM PG-RAW-90-032. For the co-elution control, buffer solution was used in place of the Cysteine stock solution.
Preparation of Positive Control and Lysine Peptide Depletion Samples and Co-elution Controls
Triplicate solutions each of the positive control and PG-RAW-90-032 stock solution were diluted with the Lysine peptide stock solution so as to prepare solutions containing 0.5 mM Lysine and 25 mM of Cinnamic Aldehyde or 25 mM PG-RAW-90-032. For the co-elution control, buffer solution was used in place of the Lysine stock solution.
Incubation
The appearance of the PG-RAW-90-032 and positive control samples in the HPLC vials was documented following preparation and then the vials placed into the autosampler of the HPLC set at 25°C for a minimum of 22 hours incubation prior to initiation of the analysis run. Prior to initiation of the run the appearance of the samples in the vials was assessed and documented again.
Analysis
The concentration of both the Cysteine and Lysine peptides in the presence of PG-RAW-90-032 and the associated positive controls was quantified by HPLC using UV detection as detailed in the chromatographic section.
Instrumentation Parameters
High performance liquid chromatograph (HPLC): Waters Alliance 2695 separation module and 2487 dual wavelength detector
Column: Agilent Zorbax SB C18, 3.5 µm, 100 x 2.1 mm
Guard column: Phenomenex AJO4286
Column temperature: 30 °C
Sample temperature: 25 °C
Mobile Phase (MP) A: 0.1% TFA in Water
Mobile Phase (MP) B: 0.085% TFA in ACN
Gradient: Time (minutes) MP A (%) MP B (%)
0 90 10
20 75 25
21 10 90
23 10 90
23.5 90 10
30 90 10
Flow rate: 0.35 mL/minute
Stroke volume 25 µL
Detector wavelength: UV, 220 nm
Injection volume: 2 µL (slow draw rate)
Run time: 30 minutes
Approximate retention time (Cysteine) 11 minutes
Approximate retention time (Lysine) 7 minutes
Results and discussion
In vitro / in chemico
Results
- Run / experiment:
- mean
- Parameter:
- other: % Overall mean peptide depletion (reactivity)
- Value:
- 0.622
- Positive controls validity:
- valid
Any other information on results incl. tables
Solubility Assessment
Solubility of PG-RAW-90-032 was achieved at a nominal concentration of 100 mM in acetonitrile.
Reactivity Assessment
All analytical acceptance criteria for each peptide run were met:
Peptide |
Standard Linearity |
Positive control depletion (%) |
Reference controls |
Test item |
|||||||
Acceptance criteria |
Cysteine |
r2>0.99 |
60.8-100 |
0.45-0.55 mM (CV <15%) |
SD <14.9% |
||||||
Lysine |
r2>0.99 |
40.2-69.0 |
0.45-0.55 mM (CV <15%) |
SD <11.6% |
|||||||
Achieved results |
Cysteine |
r2>0.999 |
71.6 |
B: 0.500 mM (CV 0.48%, n=6) |
SD 0.12% (n=3) |
||||||
Lysine |
r2>0.999 |
57.1 |
B: 0.500 mM (CV 0.66%, n=6) |
SD 0.67% (n=3) |
CV Coefficient of Variation
SD Standard deviation
The depletion of peptide in the presence of PG-RAW-90-032 was:
Peptide |
Reference Control |
Mean peak area of peptide with test item(µV.sec) |
Mean peptide depletion by |
Cysteine |
Control B: 924280 (n=6) |
912780 (n=3) |
1.24 |
Lysine |
Control B: 797070 (n=6) |
807570 (n=3) |
-1.32 |
Applying the following reactivity prediction depletion model (below), reactivity of PG-RAW-90-032 is classed as “no or minimal” and the DPRA prediction is therefore negative and is therefore predicted not to be a potential skin sensitizer.
Mean of cysteine and lysine% depletion |
Reactivity Class |
DPRA Prediction |
0%≤ mean% depletion ≤6.38% |
No or minimal reactivity |
Negative |
6.38%< mean% depletion ≤22.62% |
Low reactivity |
Positive |
22.62%< mean% depletion ≤42.47% |
Moderate reactivity |
|
42.47%< mean% depletion ≤100% |
High reactivity |
There were no co-elution peaks in either the Cysteine or Lysine assay.
Overall Achieved Depletion Values
Test item |
Cysteine peptide depletion (%) |
Lysine peptide depletion (%) |
Overall mean depletion (%) |
Reactivity class |
DPRA prediction |
||||||
PG-RAW-90-032 |
1.24 |
0.001 |
0.622 |
Minimal |
Negative |
1 For calculation of overall mean depletion a negative value counts as zero
Individual Achieved Depletion Values
Cysteine Peptide Depletion
Sample |
Peak area (µV.sec) |
Peptide concentration1(µg/mL) |
Peptide Depletion2(%) |
Mean Depletion (%) |
SD |
Positive control |
260938 |
106 |
71.8 |
71.6 |
0.16 |
263778 |
107 |
71.5 |
|||
263076 |
107 |
71.5 |
|||
PG-RAW-90-032 |
911464 |
370 |
1.39 |
1.24 |
0.12 |
913600 |
371 |
1.16 |
|||
913262 |
371 |
1.19 |
SD Standard Deviation
1 Samples prepared at a nominal concentration of 376 µg/mL (0.5 mM)
2 Calculated against a mean Reference Control (B) area of 924280 µV.sec (n=6)
Lysine Peptide Depletion
Sample |
Peak area (µV.sec) |
Peptide concentration1(µg/mL) |
Peptide Depletion2(%) |
Mean Depletion (%) |
SD |
Positive control |
336270 |
164 |
57.8 |
57.1 |
0.68 |
341481 |
167 |
57.2 |
|||
347028 |
169 |
56.5 |
|||
PG-RAW-90-032 |
808183 |
394 |
-1.39 |
-1.32 |
0.67 |
812568 |
396 |
-1.94 |
|||
801959 |
391 |
-0.613 |
SD Standard Deviation
1 Samples prepared at a nominal concentration of 388 µg/mL (0.5 mM)
2 Calculated against a mean Reference Control (B) area of 797070 µV.sec (n=6)
Applicant's summary and conclusion
- Interpretation of results:
- other: Predicted to be negative
- Remarks:
- Criteria used: EU CLP
- Conclusions:
- Solutions of PG-RAW-90-032 were successfully analyzed by the validated DPRA analytical method (Envigo analytical method FIA/M101/15) in both the Cysteine and Lysine containing synthetic peptides. With overall mean peptide depletion (reactivity) of 0.622% in the presence of the test item, PG-RAW-90-032 is only minimally reactive and is predicted by DPRA as negative and therefore unlikely to be a potential skin sensitizer based on this assay.
- Executive summary:
The purpose of this study (based on the OECD guideline for the testing of chemicals, In chemico Skin Sensitisation: Direct Peptide Reactivity Assay (DPRA), OECD/OCDE document TG 442C) was to assess the reactivity and sensitizing potential of PG-RAW-90-032.
Solutions of PG-RAW-90-032 were successfully analyzed by the validated DPRA analytical method (Envigo analytical method FIA/M101/15) in both the Cysteine and Lysine containing synthetic peptides. There was minimal reactivity of both peptides in the presence of PG-RAW-90-032. With an overall depletion of peptide of 0.622% the reactivity of PG-RAW-90-032 is henceforth classified as “no or minimal” therefore the DPRA prediction is negative. PG-RAW-90-032 is likely not to be a potential skin sensitizer based on this assay.
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.