3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooct-1-ene

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

01 Feb - 01 Mar 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

Version / remarks:

adopted Jul 2010

Deviations:

no

GLP compliance:

no

Remarks:

Since this data was to confirm the safety of the worker in the own company, it was not performed according to GLP.

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooct-1-ene

EC Number:

246-791-8

EC Name:

3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooct-1-ene

Cas Number:

25291-17-2

Molecular formula:

C8H3F13

IUPAC Name:

3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooct-1-ene

In vivo test system

Test animals

Species:

mouse

Strain:

CBA:J

Remarks:

SPF

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Charles River Japan, Inc
- Age at study initiation: 7 weeks
- Weight at study initiation: 23.2 - 24.1 g (ranges)
- Housing: 4 animals per cage were housed in polycarbonate cages (W 215 x D 320 x H 150 mm).
- Diet: Pelleted diet (MF, Oriental Yeast Co Ltd.), ad libitum
- Water: Chlorinated water, ad libitum
- Acclimation period: 13 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0 - 24.0
- Humidity (%): 50.0 - 68.0
- Air changes (per hr): 50
- Photoperiod (hrs dark / hrs light): 12/12
- IN-LIFE DATES: From: 01 Feb 2017 To: 13 Feb 2017

Study design: in vivo (LLNA)

Vehicle:

methyl ethyl ketone

Concentration:

25, 50 and 99.95%

No. of animals per dose:

4

Details on study design:

PRE-SCREEN TESTS: 2 female mice were treated by daily application of 25 μl of the test substance in concentrations of 10, 25, 50 and 99.95% to the dorsal surface of the ear, for 3 consecutive days. The body weight was recorded on Day 1 prior to dosing and on Day 6.
- Irritation: The animals were observed for local skin irritation to the application site once a day.
- Systemic toxicity: The animals were observed for signs of toxicity at least once a day.
- Ear thickness measurements: Ear thickness was measured in triplicate for each ear with the Digital micrometer on Day 1, 3 and 6.
- Erythema scores: Draize scoring system

MAIN STUDY

ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: 3H-methyl thymidine incorporation determined by β-scintillation and γ-counting
- Criteria used to consider a positive response: The mean incorporation of 3H-methyl thymidine (DPM/mouse) was calculated for the vehicle treated control group. Then, each value of incorporation of 3H-methyl thymidine in all mice was divided by the mean DPM of the vehicle treated control group to calculate stimulation index (SI). SI of vehicle treated control and test substance group were expressed as means and standard errors. The decision process regards a result as positive when SI of test substance group ≥ 3.

TREATMENT PREPARATION AND ADMINISTRATION: 25 µl of the test material was applied to the entire dorsal surface of each ear of each mouse on Days 1, 2 and 3 in concentrations of 25, 50 and 99.95% in methyl ethyl ketone. The irritation effects on the treatment site were assessed daily. On Day 6 an injection of 250 µl phosphate buffered saline (PBS) containing 20 µCi of 3H-methyl thymidine (3H-TdR) was made into the tail vein of each mouse. Approximately five hours later, the draining auricular lymph node of each ear was excised into PBS and pooled per experimental group. A single cell suspension of pooled lymph node cells was prepared by gentle mechanical disaggregation through a 200-mesh stainless steel gauze and rinsed with PBS. The precipitates were incubated for 18 h at 4°C, centrifuged, resuspended in 1 mL TCA and transferred to 10 mL scintillation fluid before β-scintillation counting.

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Results and discussion

Positive control results:

The mean SI calculated for the positive control substance (25% hexyl cinnamic aldehyde in methyl ethyl ketone) was 22.0 ± 2.2, thus meeting the reliability criteria for the LLNA (SI ≥ 3).

In vivo (LLNA)

Results

Results 1

Key result

Parameter:

SI

Value:

1.3

Variability:

0.2

Test group / Remarks:

25%

Results 2

Key result

Parameter:

SI

Value:

1.3

Variability:

0.1

Test group / Remarks:

50%

Results 3

Key result

Parameter:

SI

Value:

1.1

Variability:

0.1

Test group / Remarks:

99.95%

Cellular proliferation data / Observations:

DETAILS ON STIMULATION INDEX CALCULATION
The SI of the 25, 50 and 99.95% treatment group was 1.3, 1.3 and 1.1, respectively. None of the test substance concentrations produced as 3-fold increase in 3HTdR incorporation.

EC3 CALCULATION
None of the SI values were above 3 and it is therefore not possible to determine a EC3 concentration.

CLINICAL OBSERVATIONS
No mortality and no signs of systemic toxicity were noted in the test or control animals during the test.

BODY WEIGHTS
Body weight changes were comparable to those observed in the corresponding control group animals over the same period.

Any other information on results incl. tables

Table 1: Summary of Results

Concentration in methyl ethyl ketone (%)	Lymph node weight ± SE	Mean DPM±SE	SI ± SE
Methyl ethyl ketone	4.1±0.3	425±44	1.0±0.1
25	3.4±0.3	535±70	1.3±0.2
50	3.9±0.2	535±61	1.3±0.1
99.95	2.9±0.2	465±26	1.1±0.1
Hexyl cinnamic aldehyde (25% in methyl ethyl ketone)	9.6±0.4	9361±940	22.0±2.2

DPM = Disintegrations per minute

SE = Standard error

SI = Stimulation index

Applicant's summary and conclusion

Interpretation of results:

other: CLP/EU GHS criteria not met, no classification required according to Regulation(EC) No. 1272/2008

Conclusions:

CLP: not classified