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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vitro
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
04 Dec 2012 - 05 Feb 2013
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Performed according to GLP but no OECD guideline available yet.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013

Materials and methods

Principles of method if other than guideline:
In-vitro system for activation of antioxidant-response-element dependent genes in keratinocytes. The endpoint measurement is the up-regulation of the luciferase activity after 48 hours incubation with test substance. This up-regulation is an indicator for the activation of the Keap1/Nrf2/ARE signaling pathway.
Bauch C, Kolle SN, Ramirez T, Eltze T, Fabian E, Mehling A, Teubner W, van Ravenzwaay B, Landsiedel R, (2012), Putting the parts together: Combining in vitro methods to test for skin sensitizing potentials, Regul Toxicol Pharmacol, 63(3):489-504.
GLP compliance:
yes (incl. certificate)
Remarks:
BASF SE, Experimental Toxicology and Ecology, Ludwigshafen, Germany
Type of study:
other: part of combined in vitro sensization battery

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Fatty acids, C18-unsatd., dimers, ethoxylated (NIO AD1)
- Analytical purity: ~ 94%
- Lot/batch No.: 0007876143

In vivo test system

Test animals

Species:
other: keratinocyte cell line

Study design: in vivo (non-LLNA)

Positive control substance(s):
yes
Remarks:
Ethylene glycol dimethacrylate (EGDMA 18 µg/mL)

Results and discussion

Positive control results:
The positive control EGDMA activated the luciferase reporter 4.7a and 5.24 fold in 2 independent experiments.

Any other information on results incl. tables

Table 1: Summary of LuSens Results

 

2ndexperiment

3rdexperiment

Concentration [µg/mL]

Fold induction

Rel. viability

Fold induction

Rel. viability

VC

1.00

100.0

1.0

100.0

558*

n.d.

n.d.

1.42

80.

670*

n.d.

n.d.

1.14

76.2

804*

1.65

71.2

1.21

76.4

965*

1.67

69.7

1.29

83.1

1157*

1.43

66.5

1.22

84.9

1389*

1.36

69.8

1.01

87.6

1667*

1.06

70.5

n.d.

n.d.

2000*

0.87

79.5

n.d.

n.d.

EGDMA

4.71

87.6

5.24

103.5

LA

1.10

96.8

0.74

105.8

* Precipitation of test substance after 48 h incubation

n.d. = not determined

EGDMA = Ethylene glycol dimethacrylate (EGDMA 18 μg/mL)

LA = DL-Lactic acid (LA 450 µg/mL)

Applicant's summary and conclusion

Conclusions:
Based on the observed results it was concluded that Fatty acids, C18-unsatd., dimers, ethoxylated (NIO AD1) does not induce luciferase activity in LuSens cells under the test conditions chosen.
Executive summary:

The keratinocyte activating potential of test substance Fatty acids, C18-unsatd., dimers, ethoxylated (NIO AD1) was evaluated in the LuSens assay. For this purpose the test substance was incubated with a luciferase reporter cell line (LuSens cells) for ca. 48 hours at 37°C and ARE-dependent luciferase activity was measured in a luminometer. The test substance was soluble in 1% DMSO at a concentration of 2000 μg/mL. 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 (0.5 μg/mL up to 2000 μg/mL) and cytotoxicity was determined thereafter by MTT assay. No decrease in cell viability below 70% was observed. In the main test, test substance was used at eight final concentrations. After 48 hour exposure luciferase activity was measured in a luminometer. The test substance was concluded to exhibit a keratinocyte activating potential when the luciferase activity exceeded 1.5 fold induction with respect to the vehicle control and at concentrations that did not reduce a viability below 70%. In parallel a MTT assay was performed to assess cytotoxicity of the test substance. The LuSens showed the following results: The test substance was soluble in DMSO. The dilutions of the test substance were solutions in DMSO and suspensions in culture medium at a concentration of 558 μg/mL onward. However, after 48 hours precipitates were noticed in the samples. In summary, after 48 hours of exposure to test substance Fatty acids, C18-unsatd., dimers, ethoxylated (NIO AD1) luciferase activity in LuSens cells was was not induced at concentration affording at least 70% viability. From this it has to be concluded that Fatty acids, C18-unsatd., dimers, ethoxylated (NIO AD1) does not have a keratinocyte activating potential.