Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Toxicological information

Skin irritation / corrosion

Currently viewing:

Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2020
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2020
Report date:
2020

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Version / remarks:
2015
Qualifier:
according to guideline
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
GLP compliance:
yes (incl. QA statement)

Test material

1
Chemical structure
Reference substance name:
2,2,3,3,4,4,5,5,6,6,7,7-dodecafluoroheptan-1-ol
EC Number:
206-406-6
EC Name:
2,2,3,3,4,4,5,5,6,6,7,7-dodecafluoroheptan-1-ol
Cas Number:
335-99-9
Molecular formula:
C7H4F12O
IUPAC Name:
2,2,3,3,4,4,5,5,6,6,7,7-dodecafluoroheptan-1-ol
Test material form:
liquid

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Justification for test system used:
commercially available test system
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: epiCS® (CellSystems, Troisdorf, Germany). The model used for this study has a functional stratum comeum with an underlying layer of living cells as recommended by the test guidelines. The barrier function of the stratum comeum is adequate, as has been shown by the supplier.
- Tissue batch number(s): Cat.-No.CS-1001
- Date of initiation of testing: 26 March 2020

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: incubator (37 ± 2° C) for 20 min- exposure
- Temperature of post-treatment incubation (if applicable): 37 ± 2° C

PRE-CHECK FOR POTENTIAL OPTICAL INTERFERENCE OF THE TEST ITEM
Optical properties of the test item or its chemical action on MTT may interfere with the measurement of MTT formazan leading to a false estimate of tissue viability. The test item was therefore tested in advance for a potential direct influence on the test results not related to cytotoxic effects on tissue cells. For this pre-check the following parameters were tested:
1. Assessment of potential direct MTT-reduction of the test item
In case of a direct MTT-reduction of the test item a killed tissue control (inserts, which were killed by freezing) was used in the main assay.
2. Assessment of potential interference of colored or staining test items, which become colored after application to the tissues, with OD read out
In case of an influence of test item color on OD measurement, a color control was used in the main assay.
2.1. Assessment of the color reaction with water
2.2. Assessment of the color reaction with isopropanol

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1mg/ml
- Incubation time: 3 hours
- Spectrophotometer: EL808, Bio-Tek
- Wavelength: 570 nm

NUMBER OF INDEPENDENT TESTING RUNS / EXPERIMENTS TO DERIVE FINAL PREDICTION:
the optical density of the isopropanol-extracts of 3 insert was determined by duplicate per insert = 6 OD values.

ASSAY ACCEPTANCE CRITERIA:
The following acceptance criteria determined the validity of an assay:
- mean OD negative control is equal or greater 1.0 and equal or lower 2.8
- mean relative viability of the positive control is equal or lower 20 %
- If the mean viability of the 3 replicates is > 20% the coefficient of variation (CV) should not exceed 0.3.

PREDICTION MODEL / DECISION CRITERIA:
- The mean optical density (OD) value obtained with the test item was used to calculate the percentage of viability relative to the negative control, which is set at 100 %.
- Depending on the controls performed (see 4.4.6) the% Viability Final is calculated as follows (the values always refer to% viability)
% Viability Final = PG - (PG KC - NC KC)
- Thus the test item is identified as irritant and requiring classification according to UN GHS (Category 2 or Category 1) ifthe mean percent tissue viability after exposure and post-treatment incubation is less than or equal to 50 %. In case the test item was found to be non-corrosive (e.g. based on TG 431) and shows tissue viability after exposure and post-treatment incubation which is less than or equal to 50 %, the test item is considered to be irritant to skin in accordance with UN GHS Category 2.

The irritating potential of the test item is assessed by determination of its cytotoxic effect on an in vitro reconstructed human epidermis. The test principle is based on the MTT assay reflecting the cell viability after exposure to the topically applied test item.
All tests were performed in triplets for each time point. The test item was applied for 20 min. Cell viability was measured by the amount of MTT reduction (calculated on the basis of optical density of the negative control).
Control samples:
yes, concurrent negative control
yes, concurrent positive control
other:
Amount/concentration applied:
TEST MATERIAL APPLICATION
- Amount(s) applied: 30 µL of the test item, positive and negative control; a piece of nylon mesh was used as spreading aid

PROCEDURE FOR KILLED CONTROL
The test item was applied to two killed control tissue inserts and treated as described for the main assay (PG KC). In addition two killed control inserts were treated with 0.9 % NaCl (Negative control killed control, NC KC)

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 30 μl
- Concentration (if solution): 0.9% NaCl

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 30 µl
- Concentration (if solution): 5 % Sodium Dodecyl Sulfate (SDS)
Duration of treatment / exposure:
20 min
Duration of post-treatment incubation (if applicable):
42 hours
Number of replicates:
3

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
result for test material
Value:
2.13
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Other effects / acceptance of results:
DEMONSTRATION OF TECHNICAL PROFICIENCY:
- Reliability of the test was previously confirmed by interlaboratory validation

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative controls: yes
- Acceptance criteria met for positive control: yes
- Acceptance criteria met for variability between replicate measurements: yes

Any other information on results incl. tables

 Sample No.  test item  cell viability (%)
 1 -3  negative control (0.9% NaCl solution)  100
 6 -8  positive control (5% SDS)  1.87
 18 -20  test item  2.13

Applicant's summary and conclusion

Interpretation of results:
Category 2 (irritant) based on GHS criteria
Executive summary:

An in vitro study for predicting non-specific irritating properties of the test item was conducted according to OECD TG 439. The liquid test item was applied topically to a reconstructed human epidermis model (RhE; epiCS) in an amount of 30µL. The cell viability was determined with 2.13 % for the test item as measured by MTT conversion. It is therefore concluded that according to the criteria of this assay the test item shows irritating properties to the skin and thus requires a classification according to UN GHS (Category 2).