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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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

Administrative data

Endpoint:
skin sensitisation: in chemico
Type of information:
experimental study
Adequacy of study:
key study
Study period:
11.05.2021 - 18.05.2021
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2021

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 442C (In Chemico Skin Sensitisation Assays addressing the Adverse Outcome Pathway key event on covalent binding to proteins)
Version / remarks:
Adopted: 26 June 2020
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of study:
direct peptide reactivity assay (DPRA)
Justification for non-LLNA method:
approved alternative to animal testing

Test material

Constituent 1
Chemical structure
Reference substance name:
4-chlorobenzoic acid
EC Number:
200-805-9
EC Name:
4-chlorobenzoic acid
Cas Number:
74-11-3
Molecular formula:
C7H5ClO2
IUPAC Name:
4-chlorobenzoic acid
Test material form:
solid: particulate/powder
Details on test material:
white to yellowish

In chemico test system

Details of test system:
cysteine peptide, (Ac-RFAACAA-COOH)
lysine peptide (Ac-RFAAKAACOOH)
Details on the study design:
The DPRA is a chemistry-based assay. Nucleophile-containing synthetic peptides (cysteine peptide­Ac-RFAACAA-COOH; lysine peptide- Ac-RFAAKAA-COOH) are used to screen for skin sensitisation potential by measuring peptide depletion following incubation with allergens and non-allergens.
Synthetic heptapeptides containing either cysteine or lysine are incubated with the test item at 25 °C for 24 hours. Depletion of the peptide in the reaction mixture is measured by high pressure liquid chromatography (HPLC) using UV detection. Average peptide depletion data for cysteine and lysine are then calculated.
Vehicle / solvent:
mix DMSO:acetonitrile
Positive control:
cinnamic aldehyde

Results and discussion

Positive control results:
The mean percent peptide depletion value at 220 nm for the positive control Cinnamaldehyde is 71.0 % for the Cysteine peptide.
The mean percent peptide depletion value at 220 nm for the positive control Cinnamaldehyde is 58.6 % for the Lysine peptide.

In vitro / in chemico

Resultsopen allclose all
Group:
test chemical
Run / experiment:
run/experiment 1
Parameter:
cysteine depletion
Value:
1.1 %
At concentration:
100 mM
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Group:
test chemical
Run / experiment:
run/experiment 2
Parameter:
cysteine depletion
Value:
0.4 %
At concentration:
100 mM
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Group:
test chemical
Run / experiment:
run/experiment 3
Parameter:
cysteine depletion
Value:
2.1 %
At concentration:
100 mM
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Key result
Group:
test chemical
Run / experiment:
mean
Parameter:
cysteine depletion
Value:
1.2 %
At concentration:
100 mM
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Group:
test chemical
Run / experiment:
run/experiment 1
Parameter:
lysine depletion
Value:
0.8 %
At concentration:
100 mM
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Group:
test chemical
Run / experiment:
run/experiment 2
Parameter:
lysine depletion
Value:
0.6 %
At concentration:
100 mM
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Group:
test chemical
Run / experiment:
run/experiment 3
Parameter:
lysine depletion
Value:
0.4 %
At concentration:
100 mM
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Key result
Group:
test chemical
Run / experiment:
mean
Parameter:
lysine depletion
Value:
0.6 %
At concentration:
100 mM
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Outcome of the prediction model:
no or minimal reactivity [in chemico]
Other effects / acceptance of results:
All study acceptance criteria have been successfully met.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
4-Chlorobenzoic acid was classified as negative in the DPRA (OECD Test 442C: In Chemico Skin Sensitisation). It was assigned to reactivity class: No or minimal reactivity.
Executive summary:

The purpose of the test is to contribute to the evaluation of the skin sensitisation potential of 4-chlorobenzoic acid.

The test was performed according to OECD TG 442C, In Chemico Skin Sensitisation: Assays addressing the Adverse Outcome Pathway key event on covalent binding to proteins, Adopted: 26June 2020, Appendix I:The Direct Peptide Reactivity Assay (DPRA)

In the study, 4-chlorobenzoic acid was dissolved in dimethyl sulfoxide with acetonitrile, based on the results of the pre-experiments. The stock solution at concentration 100 mM was prepared based on a molecular weight of 156.56 g/mol.

The test item solutions were tested by incubating the samples with synthetic heptapeptides containing either cysteine or lysine at 25 °C for 24 ± 2 h. Depletion of the peptide in the reaction mixture was measured by HPLC with UV detection. Average peptide depletion data for cysteine and lysine were calculated. Cinnamaldehyde (100 mM) was used as a positive control and 1-butanol (100 mM) was used as a negative control. Each test chemical was prepared and analysed in triplicate for both peptides.

Samples were visually inspectedprior to the start of incubation andprior to HPLC analysis.

Immediately after addition of the Cysteine peptide to the solutions, there were any turbity or cloudiness, any colour changes in the vials. There were not any turbity or cloudiness, any colour changes in the vials after 24-hour incubation.

Immediately after addition of the Lysine peptide to the positive control, slight whitish turbidity was observed in the vial.The other vials were clear. The yellowish sediment on the bottom and on the wall of the vials with positive control was presented after 24-hour incubation. There was fine precipitate in the vials with test item.The other vials were clear with no color change, no turbidity and no precipitate in the vial. A phase separation was not observed for individual peptides.

All studyacceptance criteria have been successfully met.

The mean of Cysteine and Lysine percent depletion values is 0.9 %. Based on the obtained results, the prediction model the Cysteine 1:10/Lysine 1:50 was used for evaluation. The test item, 4-chlorobenzoic acid, has been found negative in the DPRA prediction; it was assigned to reactivity class “no or minimal reactivity”for both peptides.