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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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

Toxicological information

Endpoint summary

Currently viewing:

Administrative data

Description of key information

The test substance is not sensitising to skin.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
2002
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
test procedure in accordance with national standard methods with acceptable restrictions
Remarks:
sufficient for assessment with restrictions (non-guideline study)
Qualifier:
no guideline followed
Principles of method if other than guideline:
The substance was tested on fragrance sensitive volunteers.
GLP compliance:
no
Type of study:
patch test
Justification for non-LLNA method:
The data is obtained from a publication on a study performed with humans. Based on this information the LLNA criteria are not seen as requirement.
Species:
human
Strain:
not specified
Sex:
not specified
Details on test animals and environmental conditions:
- Number of subjects exposed: 218
- History of allergy or casuistics for study subject or populations: All of the probands were fragrance-sensitive.
Route:
epicutaneous, occlusive
Concentration / amount:
5 %
Route:
epicutaneous, occlusive
Concentration / amount:
5 %
No. of animals per dose:
218
Details on study design:
MAIN STUDY
A total of 218 patients with proven contact dermatitis due to fragrance materials were entered into the study. Many patients reported a personal history of hay fever, asthma or atopic dermatitis. Patch test methods and reading were according to internationally accepted criteria The patch test sites were evaluated initially at 2 - 3 days. The sites were re-examined in the majority of cases, usually between 2 and 5 days after the 1st reading.
Challenge controls:
The challenge concentrations were evaluated after treatment of 20 control subjects without evidence of fragrance allergy.
Positive control substance(s):
not required
Positive control results:
Not applicable.
Key result
Reading:
1st reading
Hours after challenge:
48
Group:
test chemical
Dose level:
5 %
No. with + reactions:
20
Total no. in group:
218
Clinical observations:
positive skin reactions
Interpretation of results:
GHS criteria not met
Conclusions:
0.9 % of the human probands were tested positive in a study on the sensitising potential of the test substance at a concentration of 5%.
Executive summary:

0.9 % of the human probands were tested positive in a study on the sensitising potential of the test substance at a concentration of 5%.

Endpoint:
skin sensitisation: in vitro
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
2015-06-01
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
new study conducted according to OECD Guideline and to GLP
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:
other: DPRA (Direct Peptide Reactivity Assay)
Details on the study design:
- Species: synthetic peptides for DPRA test system (Cysteine or Lysine containing peptides)
- TEST SYSTEM: - Source: Peptides: custom material (GenScript, Piscataway, NJ, USA and/or RS synthesis, Louisville KY, USA)
- Vehicle: Acetonitrile
- Concentration in vehicle: 100 mM; The Cysteine containing peptide was incubated with the test substance in a ration of 1:10 (0.5 mM peptide, 5 mM test substance) and the Lysine containing peptide in a ration of 1:50 (0.5 mM peptide, 25 mM test substance).
- Replicates: Three samples of the test substance were incubated with each peptide. Additionally, triplicates of the concurrent vehicle control were incubated with the peptides.
- Positive control: Ethylene glycol dimethacrylate
- Study design: The test substance was incubated with synthetic peptides for ca. 24 hours at 25 °C and the remaining non-depleted peptide concentrations were determined by HPLC with gradient elution and UV-detection at 220 nm. Calibration samples of known peptide concentration, prepared from the respective peptide stock solution used for test-substance incubation, were measured in parallel with the same analytical method. Prior to the assay the solubility of the test substance was tested.
Positive control results:
The positive control substance caused a mean peptide depletion of 30.55 %.
Run / experiment:
other: Negative control
Parameter:
other: mean cysteine-peptide depletion [%]
Value:
0
Remarks on result:
no indication of skin sensitisation
Key result
Run / experiment:
other: Test group
Parameter:
other: mean cysteine-peptide depletion [%]
Value:
0.89
Remarks on result:
no indication of skin sensitisation
Run / experiment:
other: Positive control
Parameter:
other: mean cysteine-peptide depletion [%]
Value:
55
Remarks on result:
positive indication of skin sensitisation
Run / experiment:
other: Negative control
Parameter:
other: mean lysine-peptide depletion [%]
Value:
0
Remarks on result:
no indication of skin sensitisation
Key result
Run / experiment:
other: Test group
Parameter:
other: mean lysine-peptide depletion [%]
Value:
-1
Remarks on result:
no indication of skin sensitisation
Run / experiment:
other: Positive control
Parameter:
other: mean lysine-peptide depletion [%]
Value:
6.11
Remarks on result:
no indication of skin sensitisation
Run / experiment:
other: Negative control
Parameter:
other: mean of both depletions [%]
Value:
0
Remarks on result:
no indication of skin sensitisation
Key result
Run / experiment:
other: Test group
Parameter:
other: mean of both depletions [%]
Value:
0.45
Remarks on result:
no indication of skin sensitisation
Run / experiment:
other: Positive control
Parameter:
other: mean of both depletions [%]
Value:
30.55
Remarks on result:
positive indication of skin sensitisation

The mean C-peptide depletion, caused by the test substance was determined to be 0.89 %.

The mean K-peptide depletion, caused by the test substance was determined to be -1.00 %.

Interpretation of results:
GHS criteria not met
Conclusions:
The test substance shows a minimal chemical reactivity in the DPRA under the test conditions chosen and the test result was, therefore, negative. No depletion of peptides was observed.
Executive summary:

The reactivity of the test substance towards synthetic cysteine or lysine-containing peptides was evaluated in the DPRA. The test substance was incubated with synthetic peptides for ca. 24 hours at 25 °C and the remaining non-depleted peptide concentrations were determined by HPLC. The test substance was dissolved at a 100 mM concentration in acetonitrile. Three samples of the test substance were incubated with each peptide in ratios of 1:10 (Cysteine) or 1:50 (Lysine). The following results were obtained: The mean Cysteine-peptide depletion caused by the test substance was 0.89 %, the mean Lysine-peptide depletion caused by the test substance was -1.00 %. No co-elution of test substance and peptides was noticed. Based on the observed results and applying the prediction model proposed in Gerberick et. al (2007) it was concluded that the test substance shows a minimal chemical reactivity under the test conditions chosen.

Endpoint:
skin sensitisation: in vitro
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
from 2015-06-15 to 2015-11-04
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Study conducted according to OECD Guideline and to GLP
Qualifier:
according to guideline
Guideline:
OECD Guideline 442D (In Vitro Skin Sensitisation: ARE-Nrf2 Luciferase Test Method)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of study:
other: ARE Reporter Assay (LuSens)
Details on the study design:
TEST SYSTEM:
- Species: Human transgenic keratinocyte cell line derived from HaCaT cells (LuSens)
- Source: LuSens cell line prepared in collaboration with RWTH Aachen, Germany
- Vehicle: 1 % DMSO in culture medium 3
- Concentration: A 4x concentration of the test substance with the chosen vehicle was prepared. Further concentrations were prepared by serial 1:1.2 dilutions according to the planned concentrations (master plate). In order to determine the concentrations suitable for the main experiment a pre-test (experimental conduct in accordance with GLP but without a GLP status) was performed. Cells were exposed to 9 concentrations of the test-substance preparation as provided by the sponsor (0.5 μg/mL up to 2000 μg/mL corresponding to final test substance ingredient concentrations of 0.5 μg/mL up to 1996 μg/mL taking the purity/contents of 99.8 % into account) and cytotoxicity was determined by MTT assay. The CV75 value (= estimated concentration that affords 75 % cell viability) of the test substance was determined by linear regression from the concentration response curve to be ca. 42 μg/mL (test substance as provided by the sponsor). The highest tested concentration in the 1st main experiment was 1.23 fold of the CV75 value. The additional concentrations were obtained by a 1:1.2 serial dilution series of the maximum concentration.
- No. of animals per dose: Three independent experiments were performed, in each experiment, three replicates of each test-substance concentration were tested.
- Test design: The test substance was incubated with a luciferase reporter cell line (LuSens cells) for ca. 48 h at 37 °C and antioxidant response element (ARE) dependent luciferase activity was measured in a luminometer. In order to determine the concentrations suitable for the main experiment a pre-test was performed. Cells were exposed to 9 concentrations of the test substance and cytotoxicity was determined by a MTT assay.
- Positive control: Ethylene glycol dimethacrylate
Positive control results:
The positive control substance led to a mean fold induction of luciferase activity of 7.5 and a relative viability of 120.6 %. Fold inductions of 1.50 and a relative viability of >70 % are considered as a positive result.
Run / experiment:
other: 1st experiment - test group - dose - 72 µg/mL
Parameter:
other: fold induction
Value:
1.37
Key result
Run / experiment:
other: 1st experiment - test group - dose - 1760 µg/mL
Parameter:
other: rel. viability [%]
Value:
101
Run / experiment:
other: 2nd experiment - test group - dose - 149 µg/mL
Parameter:
other: fold induction
Value:
1.34
Key result
Run / experiment:
other: 2nd experiment - test group - dose - 149 µg/mL
Parameter:
other: rel. viability [%]
Value:
72.4
Run / experiment:
other: 3rd experiment - test group - dose - 309 µg/mL
Parameter:
other: fold induction
Value:
1.91
Key result
Run / experiment:
other: 3rd experiment - test group - dose - 309 µg/mL
Parameter:
other: rel. viability [%]
Value:
35.4
Run / experiment:
other: 4th experiment - test group - dose - 309 µg/mL
Parameter:
other: fold induction
Value:
1.79
Key result
Run / experiment:
other: 4th experiment - test group - dose - 309 µg/mL
Parameter:
other: rel viability [%]
Value:
43.3
Run / experiment:
other: 5th experiment, plate 1 - test group - dose - 309 µg/mL
Parameter:
other: fold induction
Value:
1.34
Key result
Run / experiment:
other: 5th experiment, plate 1 - test group - dose - 309 µg/mL
Parameter:
other: rel. viability [%]
Value:
39.9
Run / experiment:
other: 5th experiment, plate 2 - test group - dose - 60 µg/mL
Parameter:
other: fold induction
Value:
1.53
Key result
Run / experiment:
other: 5th experiment, plate 2 - test group - dose - 60 µg/mL
Parameter:
other: rel viability [%]
Value:
84

In the 1st and 2nd experiment no keratinocyte activation was noticed. However, as no relative viability below 70 % was noticed also, the concentrations selected were too low and the experiments are not useful for evaluation. The 4th experiment is also not used for evaluation as two out of the eight concentrations tested showed relative viability above 70 %, only.

The positive and negative and vehicle control data is comparable to historic data.

Interpretation of results:
GHS criteria not met
Conclusions:
The test substance does not have a keratinocyte activating potential.
Executive summary:

The keratinocyte activating potential of the test item was evaluated in the LuSens assay. The test substance was incubated with a luciferase reporter cell line (LuSens cells) for ca. 48 h at 37 °C and antioxidant response element (ARE) dependent luciferase activity was measured in a luminometer. In order to determine the concentrations suitable for the main experiment a pre-test was performed. Cells were exposed to 9 concentrations of the test substance and cytotoxicity was determined by a MTT assay. A total of 3 valid experiments (main test) were performed. The following results were observed: After 48 hours of exposure to the test substance luciferase activity in LuSens cells was not induced in at least two consecutive concentrations with statistical significance affording at least 70 % viability in two independent experiments. Therefore it was concluded that the test item does not have a keratinocyte activating potential.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

Several studies are available which were used to assess the skin sensitizing property in a weight of evidence approach.

 

The reactivity of the test substance towards synthetic cysteine or lysine-containing peptides was evaluated in the DPRA. The test substance was incubated with synthetic peptides for ca. 24 hours at 25 °C and the remaining non-depleted peptide concentrations were determined by HPLC. The test substance was dissolved at a 100 mM concentration in acetonitrile. Three samples of the test substance were incubated with each peptide in ratios of 1:10 (Cysteine) or 1:50 (Lysine). The following results were obtained: The mean Cysteine-peptide depletion caused by the test substance was 0.89 %, the mean Lysine-peptide depletion caused by the test substance was -1.00 %. No co-elution of test substance and peptides was noticed. Based on the observed results and applying the prediction model proposed in Gerberick et. al (2007) it was concluded that the test substance shows a minimal chemical reactivity under the test conditions chosen.

 

The keratinocyte activating potential of the test item was evaluated in the LuSens assay. The test substance was incubated with a luciferase reporter cell line (LuSens cells) for ca. 48 h at 37 °C and antioxidant response element (ARE) dependent luciferase activity was measured in a luminometer. In order to determine the concentrations suitable for the main experiment a pre-test was performed. Cells were exposed to 9 concentrations of the test substance and cytotoxicity was determined by a MTT assay. A total of 3 valid experiments (main test) were performed. The following results were observed: After 48 hours of exposure to the test substance luciferase activity in LuSens cells was not induced in at least two consecutive concentrations with statistical significance affording at least 70 % viability in two independent experiments. Therefore it was concluded that the test item does not have a keratinocyte activating potential.

 

A publication (Larsen et al. 2002) is available in which a study is described that was conducted on 218 human volunteers with known allergies to fragrance materials. The test substance was applied at a concentration of 5 %. As a result, 0.9 % of the probands exhibited a positive skin reaction.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. As a result the substance is not considered to be classified as a skin sensitizer under Regulation (EC) No 1272/2008, as amended for the tenth time in Regulation (EU) No 2017/776.