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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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

Skin sensitisation

Currently viewing:

Administrative data

Endpoint:
skin sensitisation: in chemico
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
19 Oct 2015 - 22 Jan 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report date:
2016

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)
Remarks:
LANDESAMT FÜR UMWELT, WASSERWIRTSCHAFT UND GEWERBEAUFSICHT, Mainz, Germany
Type of study:
direct peptide reactivity assay (DPRA)

Test material

Constituent 1
Chemical structure
Reference substance name:
Sodium N-methyl-N-(1-oxotetradecyl)aminoacetate
EC Number:
250-151-3
EC Name:
Sodium N-methyl-N-(1-oxotetradecyl)aminoacetate
Cas Number:
30364-51-3
Molecular formula:
C17H33NO3.Na
IUPAC Name:
sodium N-methyl-N-(1-oxotetradecyl)aminoacetate
Test material form:
liquid

In chemico test system

Details on the study design:
TEST SYSTEM:
- Cysteine- (C-) containing peptide: Ac-RFAACAA-COOH
- Lysine- (K-) containing peptide: Ac-RFAAKAA-COOH

TEST SAMPLE PREPARATION:
- The test substance was prepared as a 100 mM preparation (w/v) in de-ionized water (vehicle). After short stirring the test substance was soluble in the vehicle.

CONTROLS:
- Negative control (NC): = vehicle control (VC) = de-ionized water
- Positive control (PC): Ethylene glycol dimethacrylate (EGDMA) prepared as a 50 mM preparation in de-ionized water.
- Co-elution control: Sample prepared of the respective peptide buffer and the test substance but without peptide.

EXPERIMENTAL PRODECURE:
- The test substance was dissolved in de-ionized water. Three samples of the test substance were incubated with each peptide. The C-containing peptide was incubated with the test substance in a ratio of 1:10 (0.5 mM peptide, 5 mM test substance) and the K-containing peptide in a ratio of 1:50 (0.5 mM peptide, 25 mM test substance). Additionally, triplicates of the concurrent vehicle control were incubated with the peptides. The co-elution control was prepared in the same way as the test-substance samples described above but containing the respective peptide buffer instead of peptide. The samples were prepared in suitable tubes, capped tightly and incubated at 25 °C ± 2.5 °C in the dark for 24 +/- 2 hours. Prior to HPLC analysis the samples were visually investigated for any precipitate that may have formed during the exposure period. The remaining non-depleted peptide concentration was determined thereafter by HPLC with gradient elution and UV-detection at 220 nm. In addition, 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.

MEASUREMENT OF PEPTIDE CONCENTRATIONS:
- Eluent: A: 0.1% (v/v) trifluoracetic acid (≥99%) in de-ionized water; B: 0.085% (v/v) trifluoracetic acid (≥99%) in acetonitrile

- Flow rate: 0.35 mL/min
- Wavelength: 220 and 258 nm
- HPLC: Agilent HP 1100

Results and discussion

In vitro / in chemico

Resultsopen allclose all
Key result
Run / experiment:
other: Mean value of triplicate samples
Parameter:
other: Cysteine peptide depletion
Value:
-8.52
Vehicle controls validity:
valid
Negative controls validity:
not examined
Positive controls validity:
valid
Remarks on result:
other: no indication for peptide reactivity
Key result
Run / experiment:
other: Mean value of triplicate samples
Parameter:
other: Lysine peptide depletion
Value:
0.23
Vehicle controls validity:
valid
Negative controls validity:
not examined
Positive controls validity:
valid
Remarks on result:
other: no indication for peptide reactivity
Key result
Parameter:
other: Mean of both depletions
Value:
0.11
Vehicle controls validity:
valid
Negative controls validity:
not examined
Positive controls validity:
valid
Remarks on result:
other: no indication of peptide reactivity
Other effects / acceptance of results:
ACCEPTANCE CRITERIA OF THE DPRA:
- A study is considered acceptable if the positive control causes depletion of both peptides comparable to historic data.
- The standard calibration curve should have an r² >0.99.
- The negative control (vehicle control) samples of sets A and C should be 0.50 mM +/- 0.05 mM.
- The CV of the nine vehicle controls B and C should be <15%.
- Since the mean peptide depletion for each peptide is determined from the mean of three single samples, the variability between these samples should be acceptably low (SD <14.9% for % cysteine depletion and <11.6% for % lysine depletion).

Any other information on results incl. tables

Table 3: Mean peptide depletions of cysteine, lysine and both peptides

Test item

Cysteine peptide

Lysine peptide

Mean of both depletions [%]

mean depletion [%]

SD

mean depletion [%]

SD

Test substance

-8.52*

0.73

0.23

1.58

0.11

Positive control

69.60

10.41

9.47

2.77

39.54

*a negative mean depletion [%] is considered as 0 for further calculation

SD: standard deviation

Applicant's summary and conclusion

Interpretation of results:
other: negative for peptide reactivity
Conclusions:
Based on the observed results and applying the evaluation criteria described above the test material does not exhibit peptide reactive properties in the DPRA under the test conditions chosen.