2-Propenoic acid, 2-chloro-, 3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl ester

Endpoint summary

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

8 Mar - 22 Mar 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study.

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2600 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes

Type of study:

mouse local lymph node assay (LLNA)

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Charles River France, L'Arbesle Cedex, France
- Age at study initiation: approx. 10 weeks (main study); approx. 17 weeks (preliminary study)
- Weight at study initiation: 19-23 g
- Housing: individual housing in labelled Macrolon cages (type I, height 12.5 cm) containing purified sawdust as bedding material (Woody-Clean type 3/4; Tecnilab-BMI BV, Someren, The Netherlands); during the acclimatisation period the animals were housed in polycarbonate cages (Macrolon II type, height 15 cm)
- Diet: standard animal laboratory diet (Altromin, code VRF 1, Lage, Germany), ad libitum
- Water: tap-water, ad libitum
- Acclimation period: at least 5 days before the start of the treatment

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0 +/- 3
- Humidity (%): 30-70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:

acetone/olive oil (4:1 v/v)

Concentration:

Preliminary screening test: 10, 25, 50 and 100% (v/v)
Main test: 5, 50 and 100% (v/v)

No. of animals per dose:

Preliminary screening test: 1
Main test: 5

Details on study design:

Preliminary irritation study:
A preliminary irritation study was conducted in order to select the highest test substance concentration to be used in the main study. In principle, this concentration should be well tolerated systemically by the animals and may give moderate irritation at the highest.
A series of 4 test substance concentrations were tested. The test system, procedures and techniques were identical to those used during days 1 to 3 of the main study unless otherwise specified.
4 young adult animals were selected, Each animal was treated with one concentration on two consecutive days. Approx. 4 hours after the last exposure, the skin was cleaned off residual test substance with water and the irritation was assessed. No necropsy was performed and no bodyweights were determined after termination.

Main study:
Three groups of 5 animals were treated with the test substance concentrations, one group was treated with the vehicle.
Induction (Days 1, 2 and 3):
The dorsum surface of both ears was epidermally treated (25 µL/ear) with the test substance concentration, at approx. the same time each day. The control animals were treated in the same way except that instead of the test substance the vehicle alone was administered.
Treatment (Day 6):
Each animal was injected via the tail vein with 0.25 mL of sterile phosphate buffered saline (PBS) containing 20 µCi of ³H-methyl thymidine (Amersham Pharmacia Biotech). After approx. 5 h all animals were killed by peritoneal injection with an overdose of pentobarbital. The draining (auricular) lymph node of each ear was excised. The relative size of the nodes (as compared to the normal) was estimated by visual examination and abnormalities of the nodes were recorded. The nodes were pooled for each animal in 3 mL PBS.
Tissue processing:
A single cell suspension of lymph node cells (LNC) was prepared in PBS by gentle separation through stainless steel gauze (diameter 125 µm). The LNC were washed twice and the DNA was precipitated. Precipitates were recovered by centrifugation, resuspended in 1 mL TCA and transferred to 10 mL of Ultima Gold (Packard) as the scintillation fluid.
Radioactivity measurements:
Radioactive measurements were performed using a Packard scintillation counter (1900TR). Counting time was to a statistical precision of +/- 0.2% or a maximum of 5 min whichever comes first. The scintillation counter was programmed to automatically subtract background and convert CPM to DPM.
Observation:
- Mortality/Viability: Twice daily
- Toxicity: At least once daily
- Body weights: On days 1 (pre-treatment) and 6. Body weight of the animal found dead was determined at the day of death
- Necropsy: The animal found dead was subjected to necropsy for gross macroscopic examination.
- Irritation: On day 3 (3-4 h after treatment), the skin reactions were assessed

Positive control substance(s):

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

Positive control results:

Treatment with the current positive control substance α-Hexylcinnamaldehyde, 25% in acetone:olive oil (4:1 v/v) in a reliability check resulted in a stimulation index (SI) of 16.8, thus meeting the reliability criteria for the LLNA (SI > 3).

Key result

Parameter:

SI

Value:

0.7

Variability:

+/- 0.4

Test group / Remarks:

5 %

Key result

Parameter:

SI

Value:

1.9

Variability:

+/- 0.4

Test group / Remarks:

50 %

Key result

Parameter:

SI

Value:

0.9

Variability:

+/- 0.6

Test group / Remarks:

100 %

Parameter:

SI

Remarks on result:

other: see Remark

Remarks:

The Stimulation Index expressed as the mean radioactive incorporation for each treatment group divided by the mean radioactive incorporation of the vehicle control group were as follows: 5%: 0.70+/- 0.4 50%: 1.9 +/- 0.4 100%: 0.9 +/- 0.6 0% (vehicle control): 1.0

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: Treatment with 0 (vehicle), 5, 50 and 100% test item resulted in DPM/node values of 195 +/-44 , 130+/- 49, 375 +/- 117, and 181 +/- 105 respectively.

Macroscopy:

No macroscopic abnormalities of the nodes were noted.

Body weights:

Body weight loss was noted in all animals treated with the 100% test substance concentration. The slight body weight loss noted in some animals treated with the vehicle, 5% or 50% test substance concentration was considered not toxicologically significant. Body weights and body weight gain in the other animals remained in the same range as controls over the study period.

Toxicity/mortality:

One animal treated with 100% test substance concentration was found dead on day 5. Post mortem examination of this animal revealed no abnormalities. No symptoms of systemic toxicity were observed in the animals of the main study.

Table 1: Body weights

test substance concentration (%)	animal no.	bw (g) on day 1	bw (g) on day 6
5	1	19	19
5	2	19	18
5	3	19	20
5	4	19	19
5	5	18	19
50	6	20	20
50	7	21	21
50	8	23	23
50	9	21	20
50	10	23	23
100	11	22	21
100	12	20	18
100	13	21	19
100	14	19	14
100	15	24	22
0	16	23	23
0	17	20	19
0	18	22	23
0	19	20	21
0	20	21	21

Table 2: Calculation of the Stimulation Index (SI)

group	test substance concentration (%)	mean DPM +/- SD	SI +/- SD
1	5	130 +/- 49	0.7 +/- 0.4
2	50	375 +/- 117	1.9 +/- 0.4
3	100	181 +/- 105	0.9 +/- 0.6
4	0	195 +/- 44	1.0

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl 2-chloropropenoic acid ester (CAS 96383-55-0) was tested for its skin sensitisation potential in a local lymph node assay according to OECD 429 in female CBA mice under GLP conditions (van Otterdijk, 2004). Three groups of five experimental animals were epidermally exposed to a 5%, 50% and 100% concentration, respectively of the test material on three consecutive days. The selected test substance concentrations were based on the results of a preliminary study. Five vehicle control animals were similarly treated, but with vehicle alone (Acetone/Olive oil 4:1 (v/v)). Three days after the last exposure, all animals were injected with ³H-methyl thymidine, and after five hours the draining (auricular) lymph nodes were excised. After precipitating the DNA of the lymph node cells, radioactivity measurements were done.

Body weight loss was noted in all main animals treated with 100% test substance concentration. The slight body weight loss noted in some animals of the other groups (vehicle, 5% and 50%) was considered not toxicologically significant, as the degree and incidence of weight loss was similar to that noted of the control group. Body weights and body weight gain of the other experimental animals remained in the same range as the controls over the study period. Mean DPM/animal values for the experimental groups treated with the test substance concentrations 5%, 50% and 100% were 130, 375 and 181, respectively. The mean DPM/animal value for the vehicle control group was 195. One animal treated with 100% test substance concentration was found dead on day 5. Post mortem examination of this animal did not reveal any abnormalities. No further mortality occured and no symptoms of toxicity were observed in the animals of the main study. The SI values calculated for the substance concentrations 5, 50 and 100% were 0.7, 1.9 and 0.9, respectively. Based on the results, 3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl 2-chloropropenoic acid ester is not considered a skin sensitiser under the experimental conditions of the study chosen.

Migrated from Short description of key information:

Skin sensitisation (OECD 429): not classified (no positive reactions were observed)

Justification for selection of skin sensitisation endpoint:

The reliable GLP compliant OECD Guideline study was chosen.

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information:

Migrated from Short description of key information:

no data available

Justification for selection of respiratory sensitisation endpoint:

The study is not required according to Annex VII - X of Regulation (EC) No 1907/2006.

Justification for classification or non-classification

The available data on skin sensitisation of 3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl 2-chloropropenoic acid ester do not meet the criteria for classification according to Regulation (EC) 1272/2008 or Directive 67/548/EEC, and are therefore conclusive but not sufficient for classification.