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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Experimental Starting Date: 2015-03-18; Experimental Completion Date: 2015-04-14
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Version / remarks:
and EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
yes
Remarks:
The relative humidity in the animal room was between approximately 36 - 65 % instead of 45 - 65 % for several hours. This deviation to the study plan, however, did not affect the valildity of the study.
GLP compliance:
yes (incl. certificate)
Remarks:
Hess. Ministerium für Umwelt, Energie, Landwirtschaft und Verbraucherschutz, Mainzer Straße 80 D65189 Wiesbaden
Type of study:
mouse local lymph node assay (LLNA)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: viscous liqiud
Details on test material:
Stabilited EMIM BF4:
Purity: 93.36% (EMIM+, HPLC), 86.68% (BF4-, titration); 4.89% (Stabilizer, HPLC)
Appearance: lightly yellow clear viscous liquid

In vivo test system

Test animals

Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
Number of animals for pre-test: 2 females
Number of animals for the main study: 16 females
Number of animals per group: 4 females (nulliparous and non-pregnant)
Number of test groups: 3
Number of control (vehicle) groups: 1
Age: Pre-test: 10-11 weeks (beginning of treatment)
Main study: 8-9 weeks (beginning of treatment)
Indication: The animals were distributed into the test groups at random. All animals belonging to the same experimental group were kept in one cage. In the main experiment, the animals were identified by cage number.
Acclimation: At least 5 days prior to the start of dosing under test conditions after health examination. Only animals without any visible signs of
illness were used for the study.

Study design: in vivo (LLNA)

Vehicle:
dimethylformamide
Remarks:
Purity: >= 99%
Concentration:
test item concentrations of 0, 25, 50, and 100 %
No. of animals per dose:
four female mice per concentration per dose
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)

Results and discussion

Positive control results:
Experiment performed in April 2015 (Harlan study number 1690900). Positive control substance: α-Hexylcinnamaldehyde
Vehicle: acetone:olive oil (4+1,v/v)

Test item concentration: 0% Mesurement DPM: 6228, S.I.: 1.00
Test item concentration: 5% Mesurement DPM: 12016, S.I.: 1.93
Test item concentration: 10% Mesurement DPM: 16484, S.I.: 2.65
Test item concentration: 25% Mesurement DPM: 58951, S.I.: 9.48

In vivo (LLNA)

Resultsopen allclose all
Key result
Parameter:
SI
Value:
1
Variability:
test item concentration 0%
Test group / Remarks:
4 animals
Remarks on result:
no indication of skin sensitisation based on QSAR/QSPR prediction
Key result
Parameter:
SI
Value:
0.73
Variability:
test item concentration: 25%
Test group / Remarks:
4 animals
Remarks on result:
no indication of skin sensitisation based on QSAR/QSPR prediction
Key result
Parameter:
SI
Value:
1.16
Variability:
test item concentration: 50%
Test group / Remarks:
4 animals
Remarks on result:
no indication of skin sensitisation based on QSAR/QSPR prediction
Key result
Parameter:
SI
Value:
1.45
Variability:
test item concentration 100%
Test group / Remarks:
4 animals
Remarks on result:
no indication of skin sensitisation based on QSAR/QSPR prediction

Any other information on results incl. tables

 Calculation and Results of Individual Data

Vehicle: DMF

Test item concentration %

Group

Measurement DPM

Calculation

Result

DPM-BGa)

number of lymph nodes

DPM per lymph nodeb)

S.I.

---

BG I

15

---

---

---

---

---

BG II

11

---

---

---

---

0

1

3770

3757.0

8

469.6

1.00

25

2

2753

2740.0

8

342.5

0.73

50

3

4355

4342.0

8

542.8

1.16

100

4

5476

5463.0

8

682.9

1.45

1    =  Control Group

2-4=  Test Group

a)   =  The mean value was taken from the figures BG I and BG II

b)    =  Since the lymph nodes of the animals of a dose group were pooled, DPM/node was determined by dividing the measured value by the number of lymph nodes pooled

The EC3 value could not be calculated, since all S.I.´s are below the threshold value of 3.

Viability / Mortality

No deaths occurred during the study period.

Clinical Signs

No symptoms of local skin irritation at the ears of the animals and no signs of systemic toxicity were observed during the study perio

Body Weights

The body weight of the animals, recordedprior to the first application and prior to treatment with3HTdR, was within the range commonly recorded for animals of this strain and age.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Remarks:
Migrated information
Conclusions:
The test item stabilized EMIM BF4 was not a skin sentisiser under the test conditions of this study.
Executive summary:

In the study the test item stabilized EMIM BF4 formulated in DMF was assessed for its possible skin sensitising potential.

For this purpose a local lymph node assay was performed using test item concentrations of 25, 50, and 100%. The highest concentration tested eas the highest concentration that could be achieved whilst avoiding systemic toxicity and excessive local skin irritation as confirmed by a pre-experiment.

The animals did not show any signs of systemic toxicity or local skin irritation during the course of the study and no cases of mortality were observed.

In this study Stimulation Indices (S.I.) of 0.73, 1.16, and 1.45 were determined with the test item at concentrations of 25, 50 and 100% in DMF, respectively.

The test item stabilized EMIM BF4 was not a skin sensitiser under the test conditions of this study.