1-Propanaminium, 2-hydroxy-N,N,N-trimethyl-3-[(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)thio]-, chloride (1:1)

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

28 April, 2015 - 01 May, 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)

Version / remarks:

(2014)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.40 (In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER))

Version / remarks:

(2008)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

{2-hydroxy-3-[(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)sulfanyl]propyl}trimethylazanium chloride

EC Number:

811-523-6

Cas Number:

88992-45-4

Molecular formula:

C14 H19 F13 N O S . Cl

IUPAC Name:

{2-hydroxy-3-[(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)sulfanyl]propyl}trimethylazanium chloride

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): NS-4000
- Appearance: Off-white powder (determined by WIL Research Europe)
- Storage condition of test material: At room temperature desiccated

Test animals

Species:

human

Details on test animals or test system and environmental conditions:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 36.4 - 36.7
- Humidity (%): 75 - 87

Test system

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

TEST MATERIAL
- Amounts applied: 25.1 to 28.8 mg, 25 μL Milli-Q water to moisten skin.

NEGATIVE CONTOL:
- Amount(s) applied (volume or weight with unit): 50 µL Milli-Q water

POSITIVE CONTROL
- Amount(s) applied (volume or weight with unit): 50 µL KOH
- Concentration (if solution): 8 N

Duration of treatment / exposure:

Exposure: 3 minutes and 1 hour
Post incubation period: 42 hours

Details on study design:

TEST SITE
EpiDerm Skin Model (EPI-200, Lot no.: 22226 kit N). The model consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multilayered, highly differentiated model of the human epidermis. It consists of organized basal, spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those found in vivo. The EpiDerm tissues (surface 0.6 cm²) were cultured on polycarbonate membranes of 10 mm cell culture inserts.

REMOVAL OF TEST SUBSTANCE
- Washing: phosphate buffered saline
- Time after start of exposure: 3 minutes and 1 hour

POST INCUBATION PERIOD
- 42 hours

SCORING SYSTEM:
Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) at the end of the treatment. Cell viability was calculated for each tissue as percentage of the mean of the negative control tissues.

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

other: other: percentage viability

Value:

89

Remarks on result:

other:

Remarks:

Basis: other: percentage of control. Time point: 3 minutes. Remarks: Negative control = 100%; Positive control = 14% . (migrated information)

Results 2

Irritation / corrosion parameter:

other: other: percentage viability

Value:

97

Remarks on result:

other:

Remarks:

Basis: other: percentage of control. Time point: 1 hour. Remarks: Negative control = 100%; Positive control = 7%. (migrated information)

In vivo

Irritant / corrosive response data:

The relative mean tissue viability obtained after the 3-minute and 1-hour treatments with NS-4000 compared to the negative control tissues was 89% and 97%, respectively. Because the mean relative tissue viability for NS-4000 was not below 50% after 3 minutes treatment and not below 15% after 1 hour treatment NS-4000 is considered to be not corrosive.

Other effects:

NS-4000 was checked for colour interference in aqueous conditions and possible direct MTT reduction by adding the test substance to MTT medium. Because the solutions did not turn blue / purple and a blue / purple precipitate was not observed it was concluded that NS-4000 did not interfere with the MTT endpoint.

Any other information on results incl. tables

Acceptability of the assay:

The absolute mean OD570 (optical density at 570 nm) of the negative control tissues was within the laboratory historical control data range. The mean relative tissue viability following 3-minute exposure to the positive control was 14%. The maximum inter-tissue variability in viability between two tissues treated identically was less than 22% and the maximum difference in percentage between the mean viability of two tissues and one of the two tissues was less than 13%. It was therefore concluded that the test system functioned properly.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The in vitro skin corrosion test was conducted according to OECD 431 guideline and GLP principles.
It is concluded that this test is valid and that NS-4000 is not corrosive in the in vitro skin corrosion test.

Executive summary:

In a reliable in vitro skin corrosion test using a human skin model (EpiDerm Skin Model), the influence of NS-4000 on the viability of human skin was tested. The test substance was applied directly to 0.6 cm^2 cultured skin (25.1 to 28.8 mg, in presence of 25 μL Milli-Q water). After 3 minutes and 1 hour exposure, the substance was removed and cells were cultured for 42 hours. Reliable negative and positive controls were included. The viability of the cells was tested by reduction of MTT. Survival of unexposed skin was set at 100%, the positive control had a mean cell viability of 14 and 7% whereas the test substance showed cell viability of 89 and 97%, for the 3-minute and 1 hour exposure respectively. Since the mean relative tissue viability for NS-4000 was not below 50% after the 3 -minute treatment and not below 15% after the 1-hour treatment, NS-4000 is considered to be not corrosive in the in vitro skin corrosion test.

Based on the results of this study, the substance does not need to be classified for skin corrosion according to Regulation (EC) No 1272/2008 on classification, labelling and packaging.