1H-Imidazole, reaction products with epichlorohydrin

Skin sensitisation

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Administrative data

Endpoint:

skin sensitisation: in vitro

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD Guideline for the Testing of Chemicals, Part 442E

Version / remarks:

OECD Guideline for the Testing of Chemicals, Part 442E: In Vitro Skin Sensitisation: human Cell Line Activation Test (h-CLAT) (adopted 29. July 2016)

Deviations:

not specified

GLP compliance:

yes (incl. QA statement)

Type of study:

activation of dendritic cells

Justification for non-LLNA method:

Reasons for the Choice of the THP-1 Cell Line
The OECD 442E indicates that the human monocytic leukaemia cell line, THP-1 should be used for the h-CLAT. The cells were purchased by CLS (Eppelheim, Germany).
This in vitro study is performed to assess the sensitising potential of the test item by quantifying changes in the expression level of the two cell surface markers CD86 and CD54, which are associated with the process of activation of monocytes and dendritic cells. For this purpose the human monocytic leukaemia cell line THP-1 will be used and treated with the test item for 24 h before evaluation. The measured expression levels will then be used to distinguish potential skin sensitisers from non-sensitisers.

Test material

Test material information

Constituent 1

additive 1

Test material form:

liquid

Details on test material:

- State of aggregation: not applicable
- Activation: not required
water as a additive

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: The test material is representative of the registered substance, and no significant differences in the purity profile exist
- Expiration date of the lot/batch: not relevant
- Purity test date: not relevant

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature in the dark
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle: soluble
- Reactivity of the test substance with the solvent: none

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: none
- Preliminary purification step (if any): none
- Final dilution of a dissolved solid, stock liquid or gel: none
The test material is representative of the registered substance, and no significant differences in the purity profile exist.

In vitro test system

Details on the study design:

THP-1 cells are stored in liquid nitrogen in the cell bank of LAUS GmbH to allow a continuous stock of cells, which guarantees similar parameters of the experiment and repro-ducible characteristics of the cells.
At least two weeks after thawing, a reactivity check of the cells is performed. For that, the two positive controls 2,4-dinitrochlorobenzene (DNCB) (CAS n. 97-00-7, ≥ 99% purity, test concentration: 4 µg/mL) and nickel sulphate (NiSO4) (CAS n. 10101-97-0, ≥ 99% purity, test concentration: 200 µg/mL) as well as the negative control, lactic acid (LA) (CAS n. 50-21-5, ≥ 85% purity, test concentration: 1000 µg/mL) are used. These substances as well as all additional information are given by the OECD 442E. The experimental procedure is identical to the experiments in this study. The two positive controls should produce a clearly positive result of the two surface markers CD86 and CD54. The negative control should produce a clearly negative result of both surface markers. Only cells, which suc-cessfully passed the reactivity check will be used for the study. In addition, cells will not be used longer than 2 months after thawing and do not exceed passage number 30.
After thawing the cells are cultivated in RPMI 1640 complete culture medium in cell culture flasks at 37 ± 1 °C in a humidified atmosphere with 5.0 ± 0.5 % CO2 and will be reseeded routinely every 2-3 days at a density of 0.1 to 0.2 * 106 cells/mL.

Results and discussion

Positive control results:

Detailed results not available at the moment

In vitro / in chemico

Results

Run / experiment:

other: the test item was observed to be skin sensitising.

Parameter:

other:

Remarks on result:

other: Study not available

Remarks:

the test item was observed to be skin sensitising.

Other effects / acceptance of results:

Detailed results not available at the moment

Any other information on results incl. tables

Detailed results not available at the moment

Applicant's summary and conclusion

Interpretation of results:

Category 1 (skin sensitising) based on GHS criteria

Conclusions:

the test item is skin sensitising.

Executive summary:

In a test according to OECD Guideline for the Testing of Chemicals, Part 442E: In Vitro Skin Sensitisation: human Cell Line Activation Test (h-CLAT), the test item was observed to be skin sensitising.