Amphoteric Fluorinated Surfactant

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: This study was selected as the key study because the information provided for the hazard endpoint is sufficient for the purpose of classification and labelling and/or risk assessment.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Remarks:

The study was conducted according to the guideline in effect at the time of study conduct.

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2600 (Skin Sensitisation)

Deviations:

no

Remarks:

The study was conducted according to the guideline in effect at the time the study was conducted.

GLP compliance:

yes

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Details on test material:

- Purity: 28 wt%

In vivo test system

Test animals

Species:

mouse

Strain:

other: CBA/JHsd

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Age at study initiation: approximately 9 weeks
- Weight at study initiation: 21.3 - 22.5 grams
- Housing: solid bottom cages with appropriate bedding and nestlets toys as enrichment. During quarantine, animals were housed 1 or 2 per cage. After assignment to groups, and during the in-life phase of the study, animals were housed in groups of 5.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 8 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-26ºC
- Humidity (%): 30-70%
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12-hour light/dark cycle

Study design: in vivo (LLNA)

Vehicle:

dimethyl sulphoxide

Concentration:

0% (vehicle control), 5%, 25%, 50%, 100%

No. of animals per dose:

5

Details on study design:

RANGE FINDING TESTS:
- Compound solubility: Evaluated; findings not reported

MAIN STUDY
Study Parameter            Frequency
Body Weight              Test days 0 and 5
Dosing                 Test days 0-2
Days of Rest              Test days 3-4
Injection of Radioactivity          Test day 5
Removal of Lymph Nodes          At sacrifice (test day 5)
Disintegrations per minute (dpm) data    Test day 6

ANIMAL ASSIGNMENT AND TREATMENT
- Assignment to Groups: Prior to study start using a randomly generated, computer-based algorithm such that individual pretest body weights did not vary more than 20% of the group mean.
- Daily Animal Health Observations: At least once daily to detect moribundity, mortality, and abnormal behaviour and appearance.
- Clinical Observations: Prior to administration of each dose and prior to sacrifice

TREATMENT PREPARATION AND ADMINISTRATION:
Twenty-five μL of vehicle control, test substance, or positive control (25% in vehicle control) were administered topically to the dorsum of each mouse ear for 3 consecutive days (test days 0-2). Test days 3-4 were days of rest followed by intravenous injection of 20 μCi of ³H-thymidine in PBS per mouse on test day 5.
Approximately 5 hours after the injection, animals were sacrificed by isoflurane anaesthesia followed by carbon dioxide asphyxiation, draining auricular lymph nodes were removed, and single cell suspensions were prepared. The single cell suspensions were incubated at 2-8ºC overnight. On test day 6, the single cell suspensions were counted on a beta counter and reported as disintegrations per minute (dpm).

CRITERIA USED TO CONSIDER A POSITIVE RESPONSE: A stimulation index was derived for each experimental group by dividing the mean dpm of each experimental group by the mean dpm of the vehicle control group. A stimulation index of greater than or equal to 3.0 together with consideration of dose response and, where appropriate, statistical significance were used in the determination of a positive response.

Positive control substance(s):

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

Statistics:

Significance was judged at p < 0.01. Lymph node dpm data were transformed to Log to obtain normality or homogenous variances. See Table 1 below.

Results and discussion

Positive control results:

A 25% concentration of the positive control, HCA, produced a dermal sensitization response in mice.

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: see Remark

Remarks:

No statistically significant increases in cell proliferation measurements compared to the vehicle control group were observed at any test concentration. Stimulation indices (SI)s of less than 3.0 were observed at all test concentrations of the test substance. Therefore, the EC3 value (the estimated concentration required to induce a threshold positive response, i.e., SI = 3) for the test substance under the conditions of this study was not calculable. A 25% concentration of the positive control, HCA, produced a dermal sensitization response in mice.

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: See Table 2.

Any other information on results incl. tables

No test substance-related effects on body weights were observed at any test concentration. The mean body weights of the test substance groups were similar to the negative control on test day 5. No clinical signs of toxicity were observed in the study.

Table 2 Stimulation Index Data

GROUP	MATERIAL TESTED	n	MEAN (dpm)	S.D. (dpm)	SI
1	0% Vehicle Control	5	606.15	117.73	N/A
2	5%	5	925.55	103.26	1.53
3	25%	5	873.35	462.66	1.44
4	50%	5	512.75	320.80	0.85
5	100%	5	351.35	312.15	0.58
6	25% Positive Control^a	5	2154.15	1289.80	3.55
^a Data were not included in the statistical analysis of the test substance groups.

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information

Conclusions:

The study and the conclusions which are drawn from it fulfil the quality criteria (validity, reliability, repeatability).
Not a dermal sensitizer in mice.

Executive summary:

The objective of this study was to evaluate the potential of the test substance to produce a dermal sensitization response in mice using the local lymph node assay (LLNA). Five groups of 5 female CBA/JHsd mice were dosed for 3 consecutive days with 0% (vehicle control), 5%, 25%, 50%, or 100% of test substance on both ears. Dimethylsulfoxide (DMSO) was used as the diluting vehicle. One group of 5 female mice was dosed for 3 consecutive days with 25% hexylcinnamaldehyde (HCA) in DMSO as a positive control. On test day 5 of the assay, mice received ³H-thymidine by tail vein injection and were sacrificed approximately 5 hours later. The cell proliferation in the draining auricular lymph nodes of the ears from the test substance groups was then evaluated and compared to the vehicle control group. No test substance-related effects on body weights were observed at any test concentration. The mean body weights of the test substance groups were similar to the negative control on test day 5. No clinical signs of toxicity were observed in the study. No statistically significant increases in cell proliferation measurements compared to the vehicle control group were observed at any test concentration. Stimulation indices (SIs) of less than 3.0 were observed at all test concentrations of test substance. Therefore, the EC3 value (the estimated concentration required to induce a threshold positive response, i.e., SI = 3) for the test substance under the conditions of this study was not calculable. A 25% concentration of the positive control, HCA, produced a dermal sensitization response in mice. Therefore, the LLNA test system was valid for this study with the test substance. Under the conditions of this study, the test substance did not produce a dermal sensitization response in mice. Based on these data, the test substance is not a dermal sensitizer in mice.