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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vitro
Study period:
2019
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
the study does not need to be conducted because the substance is classified as skin corrosion (Category 1, 1A, 1B or 1C)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2019

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: OECD guideline No. 442E: “In vitro skin sensitization assays addressing the Key Event on activation of dendritic cells on the adverse outcome pathway for Skin sensitization”, 25 June 2018
GLP compliance:
yes (incl. QA statement)
Type of study:
activation of dendritic cells

Test material

Constituent 1
Reference substance name:
Ethanol, 2-(2-butoxyethoxy)-, reaction products with phosphorus oxide (P2O5), compds. with N,N-dimethylcyclohexanamine
EC Number:
948-066-8
Molecular formula:
C16H36NO6P C24H52NO8P C24H53NO11P2 C32H68NO10P C8H17N H3O4P
IUPAC Name:
Ethanol, 2-(2-butoxyethoxy)-, reaction products with phosphorus oxide (P2O5), compds. with N,N-dimethylcyclohexanamine
Test material form:
liquid: viscous
Specific details on test material used for the study:
Batch No.: 0002293298
Description: Viscous colourless to yellowish liquid
Storage conditions: At room temperature
Specific requirements (handling conditions): None
Purity: Considered as 100%
Correction factor: No correction factor

In vitro test system

Details on the study design:
TEST SYSTEM
1/ Cell line
The THP-1 is an immortalized human monocytic leukemia cell line derived from an acute monocytic leukemia patient. The THP-1 cell line is obtained from ATCC (Ref: TIB-202, American Type Culture Collection, Manassas, USA) by the intermediate of LGC Standards (Molsheim, France).
The THP-1 cells are stored in a cryoprotective medium in a liquid nitrogen container. Cells will be grown using general culture procedures. They will be cultured in cRPMI medium and maintained in a humidified incubator set at 37°C, 5% CO2 and will not be allowed to exceed a cell density of 1 x 106 cells/mL or more than 30 passages.
The culture medium (cRPMI) will be composed of RPMI 1640 with 10% foetal bovine serum (FBS), 0.05 mM 2-mercaptoethanol and with penicillin (100 units/mL) and streptomycin (100 μg/mL).
During cell culturing, cell viability will be checked using trypan blue.

2/ Cell culture for testing
For testing, THP-1 cells are seeded at a density between 0.1 x 106 cells/mL and 0.2 x 106 cells/mL, and pre-cultured in culture flasks for a maximum of 72 hours. Cell will not let to exceed density of 1 x 106 cells/mL. On the day of testing, cells harvested from culture flasks are re-suspended with fresh culture medium at 2 x 106 cells/mL. Then, 500 μL of cells suspension are distributed into a 24-well flat-bottom plate (i.e. 1 x 106 cells/well).

3/ Reactivity check
Two weeks after thawing, a reactivity check is performed to qualify the cells of each working cell bank before testing. A reactivity check assay is performed by testing cell response after contact
with Lactic Acid (≥85% purity), DNCB (≥99% purity) and NiSO4 (≥99% purity) under an internal Citoxlab reference number. Results from reactivity check tests are compiled in Citoxlab France internal data, with negative and positive control data obtained during each test.

Results and discussion

In vitro / in chemico

Results
Key result
Remarks on result:
other: the test is currently ongoing.

Applicant's summary and conclusion

Interpretation of results:
other: the test is currently ongoing.