Registration Dossier

Administrative data

Description of key information

Skin irritation (OECD 404):

DSD Xi, R38; CLP Skin Irrit. 2 (>16 a.i.)

Not classified for skin irritation (≤16% a.i.)

Eye irritation (OECD 405):

DSD Xi, R41; CLP Eye Dam. 1 (>16%)

DSD Xi, R36 (≥5 ≤16% a.i.)

CLP Eye Irrit. 2 (≥1 ≤16% a.i.)

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

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

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

 


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.

Endpoint:
skin irritation / corrosion, other
Remarks:
in vivo study available
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study.
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Chemisch pharmazeutische Fabrik Dr. Karl Thomae GmbH, 88400 Biberach; SPF-breeding
- Age at study initiation: 3-5months
- Weight at study initiation: 2.0 - 2.2 kg
- Housing:individually
- Diet (e.g. ad libitum): Altromin 2123 maintenance diet for rabbits, Altromin GmbH, Lage/LIppe, ad libitum; additionally 15 g hay/day
- Water (e.g. ad libitum): tap water from automatic devices, ad libitum


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 50 ± 20
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 ml
- Concentration (if solution): ca. 30 % active
Duration of treatment / exposure:
4 hours
Observation period:
14 days
Number of animals:
3
Details on study design:
TEST SITE
- Area of exposure: dorsal area of the body
- Type of wrap if used: adhesive patch with 2.5 x 2.5 cm cellulose padding (custom-made by Fa. Beiersdorf AG, Hamburg)


REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes, with lukewarm tap water
- Time after start of exposure: 4 hours


SCORING SYSTEM: Draize scoring
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
other: 24, 48, 72 h
Score:
2.67
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
other: 24, 48, 72 h
Score:
2.67
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
other: 24, 48, 72 h
Score:
2.67
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritation parameter:
edema score
Basis:
animal #1
Time point:
other: 24, 48, 72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: Dry, rough skin surface after 72 hours, coarse scales after 72 hours and 7 days, desquamation after 7 days, reversible by day 14
Irritation parameter:
edema score
Basis:
animal #2
Time point:
other: 24, 48, 72 h
Score:
0.67
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
other: Dry, rough skin surface after 72 hours, fine scales and desquamation after 7 days, reversible by day 14
Irritation parameter:
edema score
Basis:
animal #3
Time point:
other: 24, 48, 72 h
Score:
1.33
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: Dry, rough skin surface after 72 hours, fine scales and desquamation after 7 and 14 days
Irritant / corrosive response data:
After removal of the dressing the animals showed very slight to slight edema between 30 minutes and 72 hours, and very slight to moderate erythema up to day 7. The skin surface was dry, rough and showed fine to coarse scales, with desquamation. The irritation effects were fully reversible within 14 days.

According to the criteria of Directive 67/548/EEC and the criteria of Regulation (EC) No 1272/2008 the test substance at a concentration of 30% active has to be classified as irritating to the skin (R38, CLP Skin Irrit. 2).

Interpretation of results:
irritating
Remarks:
Migrated information
Endpoint:
skin irritation: in vivo
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study with acceptable restrictions Occlusive dressing, 48h reading is missing, only 72h observation time, no individual scores reported.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
yes
Remarks:
occlusive dressing, 48h reading is missing, only 72h observation time
GLP compliance:
not specified
Species:
rabbit
Strain:
other: albino, not further specified
Type of coverage:
occlusive
Preparation of test site:
other: shaved and abraded, respectivley
Vehicle:
not specified
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5mL
- Concentration (if solution): 16%


Duration of treatment / exposure:
24h
Observation period:
24 and 72h
Number of animals:
3, non-abraded and additionally abraded on the posterior of the clipped area
Details on study design:
TEST SITE
- Area of exposure: 2x2, unit not specified
- % coverage: 10%
- Type of wrap if used: Webril patches and the entire experimental area sealed with Blenderm Surgical Tape

SCORING SYSTEM: Draize
Irritation parameter:
erythema score
Basis:
mean
Time point:
24/48/72 h
Score:
0.5
Max. score:
4
Reversibility:
fully reversible within: 72h
Remarks on result:
other: no individual scores reported
Irritation parameter:
edema score
Basis:
mean
Time point:
24/48/72 h
Score:
0.5
Max. score:
4
Reversibility:
fully reversible within: 72h
Remarks on result:
other: no individual scores reported
Irritant / corrosive response data:
The scores of the abraded skin areas were the same than for the non-abraded sites.

Coco alkyldimethyl betaines when applied as 16% solids solution to rabbit skin for 24h show only slight irritating potential after 24h that was fully reversible within 72h and has therefore not to be classified according to the criteria of EU Directive 67/548/EEC and the criteria of Regulation (EC) No 1272/2008.

Interpretation of results:
not irritating
Remarks:
Migrated information
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Referenceopen allclose all

Endpoint:
eye irritation: in vivo
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline study with acceptable restrictions. Only 10% (v/v) solution tested.
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
Only 10% (v/v) solution tested
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: David Percival Ltd., Moston, Sandbach, Cheshire, U.K., and Nottingham University, School of Agriculture, Sutton Bonington, Leicestershire, U.K.
- Age at study initiation: 12-16 weeks
- Weight at study initiation: 2.68 - 2.76 kg
- Housing: individually housed in suspended metal cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-21°C
- Humidity (%): 60-68
- Air changes (per hr): 15/h
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:
water
Controls:
other: untreated left eye served as control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1mL
- Concentration (if solution): 10%

Duration of treatment / exposure:
single instillation without washout
Observation period (in vivo):
1, 24, 48, 72h and at after 7 days
Number of animals or in vitro replicates:
3
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): No

SCORING SYSTEM: Draize

TOOL USED TO ASSESS SCORE:
light source from a standard ophthalmoscope
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1.66
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: no effects
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1.66
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1
Max. score:
2
Reversibility:
fully reversible within: 7 days
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0.33
Max. score:
2
Reversibility:
fully reversible within: 48h
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1
Max. score:
2
Reversibility:
fully reversible within: 7 days
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2.66
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24/48/72 h
Score:
1.66
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2.33
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 72h
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2.33
Max. score:
4
Reversibility:
fully reversible within: 7 days

The test material (10% v/v aqueous solution) has to be classified as irritating to the eyes according to the criteria of the EU Directive 67/548/EEC (R36) and the criteria of Regulation (EC) No 1272/2008 (Eye Irrit. 2).

Interpretation of results:
irritating
Remarks:
Migrated information
Endpoint:
eye irritation, other
Remarks:
in vivo study available
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study with acceptable restrictions. No washing, only 7 days observation period
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
No washing, only 7 days observation period
Qualifier:
according to
Guideline:
other: Leberco internal assay number 99763 (Draize method)
GLP compliance:
no
Species:
rabbit
Strain:
other: albino, not further specified
Vehicle:
unchanged (no vehicle)
Controls:
other: untreated left eyes served as controls
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
Duration of treatment / exposure:
not washed out
Observation period (in vivo):
7 days (examination every 24 hours for four days and then again on day 7)
Number of animals or in vitro replicates:
3
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): no


SCORING SYSTEM: Draize
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
1.66
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1.66
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1
Max. score:
2
Reversibility:
not reversible
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
2
Max. score:
2
Reversibility:
not reversible
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2
Max. score:
2
Reversibility:
not reversible
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2
Max. score:
3
Reversibility:
not fully reversible within: 7 days (end of observation period)
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24/48/72 h
Score:
2
Max. score:
3
Reversibility:
not reversible
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2
Max. score:
3
Reversibility:
not fully reversible within: 7 days (end of observation period)
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1.66
Max. score:
4
Reversibility:
not fully reversible within: 7 days (end of observation period)
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
3
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
not fully reversible within: 7 days (end of observation period)
Irritant / corrosive response data:
The test material caused corneal involvement and iris involvement and conjunctival irritation not clearing on the seventh day of observation.

According to the criteria of the EU Directive 67/548/EEC and the criteria of Regulation (EC) No 1272/2008 the test substance causes serious eye damage (R41 and CLP Eye Dam. 1, respectively).

Interpretation of results:
irritating
Remarks:
Migrated information
Endpoint:
eye irritation: in vivo
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study with acceptable restrictions. Only 7 days observation period
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
only 7 days observation period
GLP compliance:
no
Species:
rabbit
Strain:
other: albino, not further specified
Vehicle:
unchanged (no vehicle)
Controls:
other: untreated left eyes served as controls
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
Duration of treatment / exposure:
not washed out
Observation period (in vivo):
7 days (examination every 24 hours for four days and then again on day 7)
Number of animals or in vitro replicates:
3
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): no


SCORING SYSTEM: Draize
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
not fully reversible within: 7 days (end of observation period)
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: no effects
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: no effects
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: no effects
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24/48/72 h
Score:
1.33
Max. score:
3
Reversibility:
not fully reversible within: 7 days (end of observation period)
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1.33
Max. score:
3
Reversibility:
not fully reversible within: 7 days (end of observation period)
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0.33
Max. score:
4
Reversibility:
fully reversible within: 48h
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0.66
Max. score:
4
Reversibility:
fully reversible within: 48h
Irritant / corrosive response data:
The test material caused corneal involvement and conjunctival irritation not clearing on the seventh day of observation. The test substance induces irritation to rabbit eyes that was remaining after 7 days as slight redness in two animals and slight corneal opacity in one animal.

Table 1: Individual eye irritation scores

Rabbit No. / sex

Scoring

Cornea

Iris

Conjunctiva

Redness

Chemosis

1

24h

1

0

1

1

 

48h

1

0

1

1

 

72h

1

0

1

1

 

4 days

1

0

1

0

 

7 days

0

0

0

0

2

24h

1

0

1

0

 

48h

1

0

1

0

 

72h

1

0

1

0

 

4 days

1

0

1

0

 

7 days

0

0

1

0

3

24h

1

0

2

1

 

48h

1

0

1

1

 

72h

1

0

1

1

 

4 days

1

0

1

1

 

7 days

1

0

1

0

Conclusion: Because of the decrease of the effects over the time a reversibility after 14 or 21 days can be assumed. According to the criteria of EU Directive 67/548/EEC the test substance has not to be classified. According to the criteria of Regulation (EC) No 1272/2008 the test substance is classified as Eye Irrit. 2; H319: Causes serious eye irritation.

Interpretation of results:
irritating
Remarks:
Migrated information
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Additional information

Skin:

The most sensitive study addressing skin irritation was conducted with a 31% (according to SDS) aqueous solution of CAS 66455-29-6 (with which Betaines, C12-14 (even numbered)-alkyldimethyl was previously referred to) for 4 hours under semiocclusive conditions (Kreiling, 1994). This study demonstrated irritating properties that cause a classification as Skin Irrit. 2 according to the criteria of Regulation (EC) No 1272/2008 (CLP). Other studies performed with CAS 66455-29-6 as supplied (not further specified) for 24 hours under occlusive conditions did only indicate slight irritant properties, but not sufficient for a classification (Haynes, 1985; Haynes, 1978a, Haynes, 1978b).

One reliable study is available for CAS 68424-94-2, with exposure for 24 hours under occlusive conditions and a test material containing 16% solids (Levenstein, 1979b). The study indicated only mild irritating potential. Although not providing individual animal data, the low mean scores for the 3 tested animals for 24 and 72 hours and the complete reversibility within the observation period of 72 hours allow considering the study by Levenstein (1979b) as appropriate for assessment. It demonstrates that there is no skin irritating effect at substance concentrations up to 16%; therefore, this study was chosen as additional key study due to its importance for the determination of specific concentration limits. There is also a reliable data source available for CAS 683-10-3 (Jones and Guest, 1987). The test was performed for 4 hours under semiocclusive conditions with 10% test material concentration. This test failed to demonstrate irritating effects on the skin, either.

In addition to the information from animal tests data from several human closed patch tests with CAS 683-10-3 demonstrate skin irritation ranging from mild to strong under occlusive conditions even with concentrations as low as 1% (Basketter, 1997; Haskell Laboratory, 1963). An in vitro skin corrosion study found that Betaines, C12-14 (even numbered)-alkyldimethyl was non-corrosive.

Based on the results from the in vitro corrosion testing and available in vivo data, which show that this substance is irritating but not corrosive and only at concentrations >16%, Betaines, C12-14 (even numbered)-alkyldimethyl has been classified as Skin Irrit. 2 , H315 – Causes skin irritation at concentrations >16% .

 

Eye:

Several studies performed with CAS 66455-29-6 demonstrated only slight irritation effects on the eye, but not sufficient for a classification according to the criteria of Regulation (EC) No 1272/2008. Two studies were available in which C14 and C12 betaine were tested separately (Haynes, 1988a; Haynes, 1988b). An additional study performed at the same time on C10 betaine is also available (Haynes, 1988c). In these studies, a decreasing trend of irritation effects was observed with decreasing C-chain length. Furthermore, a number of supporting studies are available, in which no information on C-chain length was provided, all demonstrating only slight effects which were mostly reversible within the observation period of 72 hours. Even if not fully reversible within that period, the slight characteristics of the effects made it reasonable to assume full reversibility after a slightly longer observation period. However, all of these studies lacked a sufficiently detailed description of the active content of the applied substances; therefore, they are not sufficient for classification purposes and can only support the overall assessment.

Two studies for eye irritation are available for the structural analogue CAS 68424-94-2 which are both considered as key studies because they served as basis for hazard assessment and the derivation of specific concentration limits. The first one was performed with the commercial product (Levenstein, 1979c), the second one with a test material containing 16% solids (Levenstein, 1979d). In the first study, performed with the commercial product (i.e. 30% active, according to the manufacturer), iritis was observed in 2 of 3 animals exceeding the threshold values for a classification as "causing serious eye damage". This is the most sensitive study available; therefore, it will be the basis for classification and was chosen as key study. The result is a classification as Eye Dam. 1 according to Regulation (EC) No 1272/2008.

The second study for CAS 68424-94-2 was conducted with a test material containing 16% solids. It was also flagged as a key study as it is important for the setting of specific concentration limits according to Regulation (EC) No 1272/2008. In this study corneal opacity was observed which was still sufficient for a classification as Eye Irrit. 2 according to the CLP criteria. In both studies the observed effects were not or at least not fully reversible within the observation period of 7 days.

Another reliable data source is available for the structural analogue CAS 683-10-3 (actually one of the components of Betaines, C12-14 (even numbered)-alkyldimethyl). The study was performed with 10% (v/v) test material concentration (Jones and Guest, 1987), which induced irritation effects sufficient for a classification as Eye Irrit. 2 according to the criteria of Regulation (EC) No 1272/2008. The observed effects were characterised by iritis and corneal opacity. Therefore, this study was chosen as third key study as it allows to invent specific concentration limits for classification if considered together with the other two key studies for CAS 68424-94-2 (30% and 16%, Levenstein, 1979c and d, respectively).

In a worst case approach Betaines, C12-14 (even numbered)-alkyldimethyl is classified as Eye Dam. 1 and classification as irritating to eyes is assumed for concentrations between 1% and 16% as studies at concentrations 16% and 10% lead to a classification as Eye Irrit. 2.

Additionally, industrial medical surveillance data was provided for 25 workers of a production plant for CAS 683-10-3 (InfraServ GmbH & Co. KG, 2009). This routine medical check up is performed every 3 years, and the last one had been carried out in 2009. The workers were examined for disorders like irritation of the respiratory tract, the eyes, the skin, and allergic disorders or sensitisation related to contact with the test substance. In contrast to the experimental data no corresponding observations have been made in these examinations that were related to the test substance.

 

Justification for selection of skin irritation / corrosion endpoint:

Hazard assessment is based on information from experimental and read-across studies, the information provided by the different studies was used for the derivation of specific concentration limits.

Justification for selection of eye irritation endpoint:

Hazard assessment is based on read-across from three different studies for structural analogues, although studies for the substance itself are available; all were considered as key studies as the information provided by the different studies was used for the derivation of specific concentration limits.

 

Effects on skin irritation/corrosion: Irritating

 

Effects on eye irritation: corrosive

 

Justification for classification or non-classification

The available data on substances structurally closely related to Betaines, C12-14 (even numbered)-alkyldimethyl according to the criteria of Regulation (EC) No 1907/2006, Annex XI, section 1.5 showed irritating effects on rabbit and human skin and on rabbit eyes. Based on the available data the alkyl dimethyl betaines have to be classified for:

Skin irritation according to EU Directive 67/548/EEC (Xi, R38: Irritating to the skin) and Regulation (EC) No 1272/2008 (Skin Irrit. 2, H315: Causes skin irritation).

Eye irritation according to EU Directive 67/548/EEC (Xi, R41: Risk of serious damage to the eyes) and Regulation (EC) No 1272/2008 (Eye Dam. 1, H318: Causes serious eye damage).