Registration Dossier

Diss Factsheets

Toxicological information

Skin irritation / corrosion

Currently viewing:

Administrative data

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
12 July 2017 - 21 July 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Not applicable

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Version / remarks:
29 July 2016
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: EC Guideline No. 440/2008. Part B: Methods for the Determination of Toxicity and other health effects, Guideline B.40 BIS: "In Vitro Skin Corrosion: Human Skin Model Test". Official Journal of the European Union No. L142
Version / remarks:
31 May 2008
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes

Test material

Constituent 1
Reference substance name:
Reference substance 001
Cas Number:
210420-85-2
Constituent 2
Chemical structure
Reference substance name:
N-methyldidecylamine
EC Number:
230-990-1
EC Name:
N-methyldidecylamine
Cas Number:
7396-58-9
Molecular formula:
C21H45N
IUPAC Name:
N-decyl-N-methyldecan-1-amine
impurity 1
Chemical structure
Reference substance name:
Water
EC Number:
231-791-2
EC Name:
Water
Cas Number:
7732-18-5
Molecular formula:
H2O
IUPAC Name:
Dihydrogen oxide
impurity 2
Reference substance name:
Unknown impurities
IUPAC Name:
Unknown impurities
impurity 3
Chemical structure
Reference substance name:
Adipic acid
EC Number:
204-673-3
EC Name:
Adipic acid
Cas Number:
124-04-9
Molecular formula:
C6H10O4
IUPAC Name:
adipic acid
Test material form:
solid
Details on test material:
- Appearance: Slightly yellow gel
- Storage conditions: At room temperature; container flushed with nitrogen
Specific details on test material used for the study:
not applicable

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
skin obtained from plastic surgery from multiple donors
Details on animal used as source of test system:
not applicable
Justification for test system used:
Recommended test system in international guidelines (OECD and EC).
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm Skin Model
- Tissue batch number(s): 26490 (kit K and L) and 26708 (kit L and M)
- Surface: 0.6 cm^2
- On the day of receipt the tissues were kept on agarose and stored in the refrigerator.

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 3 minute exposure: room temperature, 1 hour exposure: 36.8 - 37.5°C.
- Temperature of post-treatment incubation: 37°C

REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps: the tissues were washed with phosphate buffered saline (one washing step)
- Observable damage in the tissue due to washing: no

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- Amount of MTT medium used: 300 µL
- MTT concentration: 1 mg/mL
- Incubation time: 3 hours
- Spectrophotometer: TECAN Infinite® M200 Pro Plate Reader
- Wavelength: 570 nm

NUMBER OF REPLICATE TISSUES: 4 (2 tissues for the 3 minute exposure period, 2 tissues for the 1 hour exposure period) + 2 tissue for the negative and the positive control for each exposure period

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 1

PREDICTION MODEL / DECISION CRITERIA (see table 1 in ''any other informatin on methods'')
A test item is considered corrosive in the in vitro skin corrosion test if:
a) The relative mean tissue viability obtained after 3-minute treatment compared to the negative control tissues is decreased below 50%.
b) In addition, a test item considered non-corrosive (viability = 50%) after the 3-minute treatment is considered corrosive if the relative tissue viability after 1-hour treatment with the test item is decreased below 15%.

A test item is considered non corrosive in the in vitro skin corrosion test if:
a) The relative mean tissue viability obtained after the 3-minute treatment compared to the negative control tissues is not decreased below 50%.
b) In addition, the relative tissue viability after the 1-hour treatment is not decreased below 15%.

ACCEPTABILITY CRITERIA:
a) The absolute mean OD570 of the two tissues of the negative control should reasonably be within the acceptance limits of OECD 431 (lower acceptance limit =0.8 and upper acceptance limit =2.8)
b) The mean relative tissue viability following 1-hour exposure to the positive control should be <15 %.
c) In the range 20 - 100% viability, the Coefficient of Variation (CV) between tissue replicates should be = 30%.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount applied: an excess amount (undiluted)

NEGATIVE CONTROL
- Amount applied: 50 µL

POSITIVE CONTROL
- Amount applied: 50 µL (8N KOH)
Duration of treatment / exposure:
3 minutes and 1 hour
Duration of post-treatment incubation (if applicable):
3 hours
Number of replicates:
4 in total; 2 per exposure period

Test animals

Details on test animals or test system and environmental conditions:
not applicable

Test system

Type of coverage:
open
Preparation of test site:
not specified
Vehicle:
unchanged (no vehicle)
Amount / concentration applied:
Liquid test item applied directly on top of the tissue
Duration of treatment / exposure:
3 minute exposure and 1 hour exposure
Observation period:
not applicable
Number of animals:
not applicable
Details on study design:
not applicable

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
% tissue viability
Remarks:
mean of 2 replicates
Run / experiment:
3-minute exposure period; experiment 1
Value:
53
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks:
6.7%
Remarks on result:
other: CV between replicates: 16%
Irritation / corrosion parameter:
% tissue viability
Remarks:
mean of 2 replicates
Run / experiment:
3-minute exposure period; experiment 2
Value:
39
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks:
6.3%
Remarks on result:
other: CV between replicates: 10%
Irritation / corrosion parameter:
% tissue viability
Remarks:
mean of 2 replicates
Run / experiment:
1-hour exposure period
Value:
36
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks:
9.6%
Remarks on result:
other: CV between replicates: 23%
Other effects / acceptance of results:
- The individual viabilities of the tissues treated with test item were spread over 2 categories (48% and 57%) therefore the 3-minute exposure period was repeated. The repeated experiment showed individual tissue viabilities of 37% and 42% and was therefore considered acceptable and used to determine the final conclusion.

- OTHER EFFECTS:
- Visible damage on test system: no
- Direct-MTT reduction: no
- Colour interference with MTT: no

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes, the absolute mean OD570 was between the acceptance limits (i.e., 1.971 for the 3-minute exposure and 1.744 for the 1-hour exposure).
- Acceptance criteria met for positive control: yes, the mean relative tissue viability following 1 hour exposure was <15% (i.e., 9.6%).
- Acceptance criteria met for variability between replicate measurements: yes, CV between tissue replicates was =30% (i.e., 10% for the 3 minute exposure periods and 23% for the 1 hour exposure period).

In vivo

Irritant / corrosive response data:
not applicable
Other effects:
not applicable

Any other information on results incl. tables

Table 2 Individual OD measurements at 570 nm

 

3-minute application (OD570)

       A               B

1-hour application (OD570)       A               B

Negative control

OD570measurement 1

OD570measurement 2

OD570measurement 3

 

 

1.6481

1.7265

1.7199

1.8792

1.6399

1.7571

1.6803

1.8309

1.7038

1.7773

1.7232

1.8794

1-Decanaminium, N-decyl-N,N-dimethyl-, hexanedioate (2:1)

OD570measurement 1

OD570measurement 2

OD570measurement 3

 

 

0.8198

0.9787

0.5830

0.7528

0.8592

1.0003

0.5961

0.7578

0.8449

1.0043

0.6021

0.7686

Positive control

OD570measurement 1

OD570measurement 2

OD570measurement 3

 

 

0.1488

0.1510

0.2237

0.1978

0.1585

0.1482

0.2186

0.1980

0.1607

0.1536

0.2237

0.1954

 Repeat experiment

3-minute application (OD570)

       A               B

Negative control

OD570measurement 1

OD570measurement 2

OD570measurement 3

 

1.9161

2.0235

2.0228

2.0685

2.0073

2.0446

1-Decanaminium, N-decyl-N,N-dimethyl-, hexanedioate (2:1)

OD570measurement 1

OD570measurement 2

OD570measurement 3

 

0.7352

0.8733

0.7976

0.8560

0.8016

0.8579

Positive control

OD570measurement 1

OD570measurement 2

OD570measurement 3

 

0.1424

0.1780

0.1579

0.1833

0.1567

0.1826

OD = Optical density; Duplicate exposures are indicated by A and B.

Applicant's summary and conclusion

Interpretation of results:
Category 1 (corrosive) based on GHS criteria
Conclusions:
The results of a skin corrosion test, performed according to OECD guideline 431, showed that 1-Decanaminium, N-decyl-N,N-dimethyl-, hexanedioate (2:1) was corrosive to the skin (tissue viability of 39% after 3-minute exposure). The test substance is therefore classified as category 1 according to GHS and according to Regulation (EC) 1272/2008.
Executive summary:

The results of a skin corrosion test, performed according to OECD guideline 431, showed that 1-Decanaminium, N-decyl-N,N-dimethyl-, hexanedioate (2:1) was corrosive to the skin (tissue viability of 39% after 3-minute exposure). The test substance is therefore classified as category 1 according to GHS and according to Regulation (EC) 1272/2008.