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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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 corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
11 February 2019 - 14 February 2019
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2019
Report date:
2019

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Deviations:
no
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
Betaines, C12-14 (even numbered)-alkyldimethyl
EC Number:
931-700-2
Cas Number:
66455-29-6
Molecular formula:
C16H33NO2 C18H37NO2
IUPAC Name:
Betaines, C12-14 (even numbered)-alkyldimethyl
Test material form:
solid
Remarks:
The test item was prepared by drying of the manufactured liquid alkyldimethyl betaine substance.
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: BLBL016270
- Expiration date of the lot/batch: 10 January 2020
- Purity test date: Not specified

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature
- Stability under test conditions: Stable
- Solubility and stability of the test substance in the solvent/vehicle: Soluble in water and ethanol
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: Not specified

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
other: Normal Human Epidermal Keratinocytes (NHEKs) are isolated from the epidermis of neonatal foreskin (NHEK-CRY-NEO) or adult (breast or abdominal) skin (NHEK-CRY-AD) and cryopreserved.
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDermTM (EPI-200) Reconstructed Human Epidermis
- Tissue batch number(s): 28683
- Delivery date: 12 February 2019
- Date of initiation of testing: 13 February 2019

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37°C

REMOVAL OF TEST MATERIAL AND CONTROLS
- Not specified

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- Incubation time: 3 minutes; 1 hour
- Spectrophotometer: BMG LabTech FluoStar Optima
- Wavelength: 570 nm
- Filter: No filter
- Linear OD range of spectrophotometer:

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: 2.047 ± 0.057 hrs (PASS)
- Barrier function: 4.79 hrs (PASS)
- Morphology: PASS
- Contamination: No contamination reported (PASS)

NUMBER OF REPLICATE TISSUES: 3
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied: 25 mg

NEGATIVE CONTROL
- Amount(s) applied: 50 µL

POSITIVE CONTROL
- Amount(s) applied: 50 µL
- Concentration (if solution): 8N
Duration of treatment / exposure:
3 minutes; 1 hour
Number of replicates:
3

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
% tissue viability
Remarks:
Average % tissue viability in triplicate tissues
Run / experiment:
3 minute endpoint
Value:
100.67
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Remarks:
Average viability 100.00%
Positive controls validity:
valid
Remarks:
Average viability 5.82%
Irritation / corrosion parameter:
% tissue viability
Remarks:
Average % tissue viability in triplicate tissues
Run / experiment:
1 hour endpoint
Value:
25.68
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Remarks:
Average viability 100.00%
Positive controls validity:
valid
Remarks:
Average viability 5.75%
Other effects / acceptance of results:
OVERALL RESULT:
The test item neither reduced the viability to below 50% after 3 mins nor below 15% after 1h and therefore should be considered as Non-Corrosive.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The mean OD570 of the negative control tissues must be ≥0.8. Actual values: 2.41 (3 min); 2.08 (1 hr). PASS.
- Acceptance criteria met for positive control: The mean of the positive control relative percentage viability, after 1hour exposure must be < 15% of the mean of the negative control. Actual value: 5.75. PASS.
- Acceptance criteria met for variability between replicate measurements: In the range between 20% and 100% viability, the coefficient of variation (CV) is an additional acceptance criterion. It should not exceed 0.3 (30%). PASS.

Any other information on results incl. tables

Cell viability measurements after 3 minutes of application.

3 Minute Endpoint

NC

PC

TA2

Tissue 1

Aliquot 1 OD

2.38

0.14

2.12

Aliquot 2 OD

2.38

0.13

2.14

Tissue 2

Aliquot 1 OD

2.50

0.22

2.45

Aliquot 2 OD

2.57

0.22

2.49

Tissue 3

Aliquot 1 OD

2.31

0.06

2.65

Aliquot 2 OD

2.30

0.07

2.69

All Tissues

Average OD

2.41

0.14

2.42

Average Viability

100.00

5.82

100.67

Average % SD

4.01

2.63

9.25

Average % CV

4.01

45.10

9.18

NC: negative control (H2O), PC: Positive control (KOH 8N), TA2: of Betaines, C12-14 (even numbered)-alkyldimethyl.

*CV value of the PC is above 30% (45.10%) however, as the percentage of viability is below 20%

(5.82%) acceptance criterion 3 is met.

 

 

Cell viability measurements after 1 hour of application.

1 Hour Endpoint

NC

PC

TA2

Tissue 1

Aliquot 1 OD

2.18

0.12

0.34

Aliquot 2 OD

2.20

0.11

0.33

Tissue 2

Aliquot 1 OD

2.08

0.21

0.54

Aliquot 2 OD

2.03

0.23

0.55

Tissue 3

Aliquot 1 OD

1.95

0.02

0.71

Aliquot 2 OD

2.04

0.03

0.73

All Tissues

Average OD

2.08

0.12

0.53

Average Viability

100.00

5.75

25.68

Average % SD

4.01

3.76

7.62

Average % CV

4.01

65.52

29.67

NC: negative control (H2O), PC: Positive control (KOH 8N), TA2: of Betaines, C12-14 (even numbered)-alkyldimethyl.

*CV value of the PC is above 30% (65.52%) however, as the percentage of viability is below 20%

(5.75%) acceptance criterion 3 is met.

 

Evaluation of the results- Results were checked against the following acceptance criteria:

 

 

Actual values

Pass/Fail

Acceptance criterion 1

The mean OD570 of the negative control tissues must be ≥0.8.

3 min = 2.41

1 hr = 2.08

Pass

Acceptance criterion 2

The mean of the positive control relative % viability after 1 hr exposure must be <15% of the mean of the negative control.

 

5.75

Pass

Acceptance criterion 3

In the range between

20% and 100% viability, the coefficient of

variation (CV) is an

additional acceptance

criterion.It should not

exceed 0.3(30%).

 

CV

3 min

1 hr

NC

4.01

4.01

PC

45.10

65.52

TA2

9.18

29.67

Pass

*Note that the CV values of the PC after 3 min and 1hr were above 30% (45.10% and 65.52% respectively). However, as the percentage of cell viability was below 20% (5.82% and 5.75% respectively), acceptance criterion 3 was met.

 

Results Summary

 

Test item

Test Item

ID

Viability

after

3 minutes

application

Viability ≥

50% after

3 minutes

(Yes/No)

Viability

after 1h

application

(% to

negative

control)

Viability ≥

15% after 1

hour (Yes/

No)

Corrosive

(C) / Non

corrosive

(NC)

Betaines, C12-14 (even numbered)-alkyldimethyl

TA2

100.67

Yes

25.68

Yes

Non-Corrosive

 


Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
In an in vitro skin corrosion study according to OECD TG 431 (RHE method), Betaines, C12-14 (even numbered)-alkyldimethyl was found to be non-corrosive to skin.
Executive summary:

The potential of Betaines, C12-14 (even numbered)-alkyldimethyl to cause corrosion to skin was assessed in an in vitro study by the reconstructed human epidermis (RHE) test method, OECD TG 431. The test material, dried to solid form, was applied neat to unchanged human-derived epidermal keratinocytes for 3 minutes or 1 hour. The test item neither reduced the viability to below 50% after 3 mins nor below 15% after 1 hour and therefore should be considered as Non-Corrosive.