Registration Dossier

Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Skin irritation / corrosion.

The skin irritation / corrosion potential of the test substance, bis(2 -hydroxyethyl)oleylmethylammonium chloride, CAS Number 18448-65-27, EC Number 242-332-02, was investigated in vitro according to OECD Test Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis Test Method).

Under the conditions of this study after 3 min and 1h exposure on the surface of EpiDermTM reconstituted human epidermis, the viability of the tissues was assessed and compared to a negative control.

The percentage of viability obtained was 81.4% after 3min and 89.0% after 1h and therefore:

Bis (2-Hydroxyethyl) Oleyl Methyl Ammonium Chloride (CAS 18448-65-2, EC 242-332-0) was classified as Non Corrosive to human skin.

In addition to the study described above read-across data is presented as follows :

The skin irritation / corrosion potential of the substance, Quaternary ammonium compounds, C12-18-alkylbis(hydroxyethyl)methyl, chlorides, CAS Number 71808 -53 -2, EC Number 276 -038 -9, was investigated in a study conducted according to OECD Guideline 404 (Acute Dermal Irritation / Corrosion) / EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion) / EPA OPPTS 870.2500 (Acute Dermal Irritation).

This substance is considered to be close enough in structural integrity to the target substance, bis(2 -hydroxyethyl)oleylmethylammonium chloride, CAS Number 18448 -65 -2, EC Number 242 -332 -0, so as to justify valid read-across.

Under the conditions of the present study, the single dermal application of the test item C12-18-alkylbis(hydroxyethyl)methyl, chloride (77% in isopropyl alcohol) to one rabbit at a dose of 0.5 g for 3 min exposure showed irritant effects, for 1 hour and 4 hours exposure showed corrosive effects which were not fully reversible within the observation period.

Eye irritation

The study does not need to be conducted because the substance is classified as skin irritation, leading to classification as eye irritation (Category 2).

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP Guideline Study
Justification for type of information:
Read-across data is presented from quaternary ammonium compounds, C12-18-alkylbis(hydroxyethyl)methyl, chlorides, CAS Number 71808-53-2, EC Number 276-038-9. This substance is considered to be close enough in structural integrity to the target substance, bis(2 -hydroxyethyl)oleylmethylammonium chloride, CAS Number 18448 -65 -2, EC Number 242 -332 -0, so as to justify valid read-across.
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.2500 (Acute Dermal Irritation)
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
Name: C12-18-alkylbis(hydroxyethyl)methyl, chloride (77% in isopropyl alcohol)
CAS No.: 71808-53-2 (old CAS No. 70750-47-9)
Chemical Name: C12-18-alkylbis(hydroxyethyl)methyl, chloride
Batch No.: S001314
Physical state at RT: liquid
Colour: amber
Active Components: 77.3%
Storage: at room temperature
Expiry Date: 28.04.2018
Species:
rabbit
Strain:
New Zealand White
Details on test animals or test system and environmental conditions:
- Semi barrier in an air-conditioned room
- Temperature: 18 +/- 3 °C (recommendations of TVT, GV-SOLAS)
- Relative humidity: 55 +/- 10%
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: at least 10 x / hour
- Free access to autoclaved hay and to Altromin 2123 maintenance diet for rabbits (lot no. 0700), rich in crude fibre
- Free access to tap water (drinking water, municipal residue control, microbiological controls at regular intervals)
- Certificates of food, water and bedding are filed at BSL BIOSERVICE
- Housed in ABS - plastic rabbit cages, floor 4200 cm2
- Adequate acclimatisation period (at least 5 days) under laboratory conditions
Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
0.5 ml
Duration of treatment / exposure:
As the test item was suspected to be corrosive, three patches were applied to one animal. The exposure periods were 3 min, 1 hour and 4 hours.
At the end of the exposure period, the residual test item was removed with tap water.
Observation period:
14 days after the patch removal
Number of animals:
1
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
48 h
Score:
2
Remarks on result:
other: 3 min exposure - 24h to 48h
Irritation parameter:
edema score
Basis:
animal #1
Time point:
48 h
Score:
1
Remarks on result:
other: 3 min exposure - 24h to 48h
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
3
Remarks on result:
other: 1h exposure - 24h to 72h
Irritation parameter:
edema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2
Remarks on result:
other: 1h exposure - 24h to 72h
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
4
Remarks on result:
other: 4h exposure - 24h to 72h
Irritation parameter:
edema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2
Remarks on result:
other: 4h exposure - 24h to 72h
Irritant / corrosive response data:
Initial test in animal no. 1: The examination of the test site immediately after the patch removal revealed erythema grade 1 and oedema grade 1 for the 3 min exposure, erythema grade 1 and oedema grade 2 for the 1 hour exposure and erythema grade 2 and oedema grade 3 for the 4 hour exposure.
The subsequent clinical observations showed irritant effects for the 3 min exposure and corrosive effects for the 1 hour and 4 hours exposure, which were not reversible within the observation period of 14 days.
Irritant effects were observed on the intact skin of one female rabbit (strain NZW) after a contact time of 3 min (Tables), corrosive effects were observed on the intact skin of one female rabbit after a contact time of 1 hour and 4 hours. These signs of irritation / corrosion were not reversible within the observation period of 14 days.

Dermal Irritation Evaluation

Exposure Time

Application
Site

Irritation (hours after patch removal)

1 hour

24 hours

48 hours

72 hours

T

C

T

C

T

C

T

C

3 min

Erythema

1

0

2

0

2

0

2

0

Oedema

1

0

1

0

1

0

1

0

1 hour

Erythema

1

0

3

0

3

0

3

0

Oedema

2

0

2

0

2

0

2

0

4 hours

Erythema

3

0

4

0

4

0

4

0

Oedema

3

0

2

0

2

0

2

0

Exposure Time

Application
Site

Irritation (days after patch removal)

4 days

5 days

6 days

7 days

T

C

T

C

T

C

T

C

3 min

Erythema

2

0

2

0

2

0

1

0

Oedema

1

0

1

0

0

0

0

0

1 hour

Erythema

3

0

3

0

3

0

2

0

Oedema

2

0

2

0

0

0

0

0

4 hours

Erythema

4

0

4

0

4

0

4

0

Oedema

2

0

2

0

0

0

0

0

Exposure Time

Application
Site

Irritation (days after patch removal)

8 days

9 days

10 days

11 days

T

C

T

C

T

C

T

C

3 min

Erythema

1

0

1

0

0

0

0

0

Oedema

0

0

0

0

0

0

0

0

1 hour

Erythema

2

0

2

0

1

0

1

0

Oedema

0

0

0

0

0

0

0

0

4 hours

Erythema

4

0

4

0

4

0

3

0

Oedema

0

0

0

0

0

0

0

0

Exposure Time

Application
Site

Irritation (days after patch removal)

12 days

13 days

14 days

 

T

C

T

C

T

C

 

3 min

Erythema

1

0

1

0

0

0

 

Oedema

0

0

0

0

0

0

 

1 hour

Erythema

2

0

2

0

1

0

 

Oedema

0

0

0

0

0

0

 

4 hours

Erythema

4

0

4

0

4

0

 

Oedema

0

0

0

0

0

0

 

Other findings:

Individual Systemic and Local Findings – 3 min exposure

Time after
Patch Removal

Systemic Findings

Specific Local Findings (apart from erythema/oedema)

Comments

1 hour

nsf

nsf

-

24 hours

nsf

nsf

-

48 hours

nsf

nsf

-

72 hours

nsf

nsf

-

4 days

nsf

eschar, superficial cleft

-

5 days

nsf

eschar, superficial cleft

-

6 days

nsf

eschar, superficial cleft

-

7 days

nsf

eschar, desquamation

-

8 days

nsf

eschar, desquamation

-

9 days

nsf

eschar, desquamation

-

10 days

nsf

eschar, desquamation

-

11 days

nsf

eschar, desquamation

-

12 days

nsf

eschar, desquamation

-

13 days

nsf

dequamation

-

14 days

nsf

dequamation

-

Individual Systemic and Local Findings – 1 hour exposure

Time after
Patch Removal

Systemic Findings

Specific Local Findings (apart from erythema/oedema)

Comments

1 hour

nsf

nsf

-

24 hours

nsf

eschar

-

48 hours

nsf

eschar

-

72 hours

nsf

eschar

-

4 days

nsf

eschar, cleft

-

5 days

nsf

eschar, cleft

-

6 days

nsf

eschar, cleft

-

7 days

nsf

eschar, desquamation

-

8 days

nsf

eschar, desquamation

-

9 days

nsf

eschar, desquamation

-

10 days

nsf

eschar, desquamation

-

11 days

nsf

eschar, desquamation

-

12 days

nsf

eschar, desquamation

-

13 days

nsf

eschar, desquamation

-

14 days

nsf

eschar, desquamation

-

Individual Systemic and Local Findings – 4 hours exposure

Time after
Patch Removal

Systemic Findings

Specific Local Findings (apart from erythema/oedema)

Comments

1 hour

nsf

nsf

-

24 hours

nsf

eschar

-

48 hours

nsf

eschar

-

72 hours

nsf

eschar

-

4 days

nsf

eschar

-

5 days

nsf

eschar

-

6 days

nsf

eschar

-

7 days

nsf

eschar

-

8 days

nsf

eschar

-

9 days

nsf

eschar

-

10 days

nsf

eschar

-

11 days

nsf

eschar

-

12 days

nsf

eschar

-

13 days

nsf

eschar

-

14 days

nsf

eschar

-

Body Weight Develompent:

 

Animal No. 1

Start of Study
(weight in kg)

2.5

72 Hours after Application (weight in kg)

2.4

End of Study
(weight in kg)

2.7

Interpretation of results:
Category 1B (corrosive) based on GHS criteria
Conclusions:
Under the conditions of the present study, the single dermal application of the test item C12-18-alkylbis(hydroxyethyl)methyl, chloride (77% in isopropyl alcohol) to one rabbit at a dose of 0.5 g for 3 min exposure showed irritant effects, for 1 hour and 4 hours exposure showed corrosive effects which were not fully reversible within the observation period.
Executive summary:

The skin irritation / corrosion potential of the test substance, Quaternary ammonium compounds, C12-18-alkylbis(hydroxyethyl)methyl, chlorides, CAS Number 71808 -53 -2, EC Number 276 -038 -9, was investigated in a study conducted according to OECD Guideline 404 (Acute Dermal Irritation / Corrosion) / EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion) / EPA OPPTS 870.2500 (Acute Dermal Irritation).

This substance is considered to be close enough in structural integrity to the target substance, bis(2 -hydroxyethyl)oleylmethylammonium chloride, CAS Number 18448 -65 -2, EC Number 242 -332 -0, so as to justify valid read-across.

The study was assigned a reliability score of 1 in accordance with the criteria for assessing data quality set forth by Klimisch et al. (1997).

Under the conditions of the present study, the single dermal application of the test item C12-18-alkylbis(hydroxyethyl)methyl, chloride (77% in isopropyl alcohol) to one rabbit at a dose of 0.5 g for 3 min exposure showed irritant effects, for 1 hour and 4 hours exposure showed corrosive effects which were not fully reversible within the observation period.

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
7th February 2018 - 1st March 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Version / remarks:
XCellR8 SOP L0048 ‘EpiDermTM In Vitro Reconstructed Human Epidermis Skin Corrosion Test’, is based on the current MatTek protocol (MK-24-007-0024, 07/11/2014) using the MatTek Corporation EpidermTM reconstructed tissue model: EPI-200. This method was endorsed in 2002 by National Co-ordinators of OECD Test Guideline Programme (WNT) originally adopted in 2004 and updated in 2013 and 2015.
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Vehicle:
unchanged (no vehicle)
Details on test system:
The reconstructed human epidermal model EpidermTM (EPI-200 MatTek Corporation) consists of normal human-derived epidermal keratinocytes which have been cultured to form a multi-layered highly differential model of the human epidermis. It consists of organised 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.

MatTek’s EpiDermTM model has been extensively characterised for multiple parameters including morphology, tissue viability, skin barrier function and sterility (for information see www.mattek.com). QC results for the specific lot of models received (Lot# 25883) were checked in-house for MatTek acceptance ranges with the following outcome :
 Morphology - PASS
 Tissue viability - PASS
 Skin barrier function (ET50 value for 1% Triton X-100) where ET50 is the time taken for 1% Triton X-100 to reduce the viability of the skin model to 50% relative to the negative control)- PASS
 Sterility testing showed no contamination during long term antibiotic and antimycotic free culture- PASS

Single topical application of 25μl H20 plus nominal 25mg/50μl neat test or reference items to the surface of the EpiDermTM skin model for 3 and 60 minutes at 37°C, 5% CO2 ≥95% RH (Relative Humidity), prior to the MTT endpoint; three tissues per condition (n=3). Test item was applied using a pin applicator.

Main Test-overview
Day 1: On the day of receipt, EpiDermTM tissues were kept at 4°C till the next day.
Day 2: EpiDermTM tissues were pre-incubated 1h at 37°C, 5% CO2, 95% RH (Relative Humidity).
Day 2: Exposure to and removal of neat test and reference items in triplicates (n=3) for 3 and 60 minutes; MTT viability testing.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
25 mg/50μl neat test or reference items
Duration of treatment / exposure:
3 and 60 minutes at 37°C, 5% CO2 ≥95% RH (Relative Humidity),
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
1, 3 minutes
Value:
81.4
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: 81.4% viability after 3 minutes
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
2, 60 minutes
Value:
89
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: 89% viability after 60 minutes
Other effects / acceptance of results:
The test item did not reduce the viability below 50% after 3 min nor below 15% after 1h and should be considered as non-corrosive.
Interpretation of results:
GHS criteria not met
Conclusions:
Bis (2-Hydroxyethyl) Oleyl Methyl Ammonium Chloride (CAS 18448-65-2, EC 242-332-0) evaluated for skin corrosion following OECD Guideline TG 431 and using EpiDermTM tissue model was non-corrosive.
Executive summary:

The skin irritation / corrosion potential of the test substance, bis(2 -hydroxyethyl)oleylmethylammonium chloride, CAS Number 18448-65-27, EC Number 242-332-02, was investigated in vitro according to OECD Test Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis Test Method).

The study was conducted in accordance with:

  • The OECD Principles on Good Laboratory Practice [GLP) (ENV/MC/CHEM(98)17] (Jan 1998).
  • UK Statutory Instrument 1999 Number 3106: The Good Laboratory Practice Regulations.
  • UK Statutory Instrument 2004 Number 994: The Good Laboratory Practice (Codification Amendments etc.) Regulations.

The study was assigned a reliability score of 1 in accordance with the criteria for assessing data quality set forth by Klimisch et al. (1997).

Under the conditions of this study after 3 min and 1h exposure on the surface of EpiDermTM reconstituted human epidermis, the viability of the tissues was assessed and compared to a negative control.

The percentage of viability obtained was 81.4% after 3min and 89.0% after 1h and therefore:

Bis (2-Hydroxyethyl) Oleyl Methyl Ammonium Chloride (CAS 18448-65-2, EC NUMBER 242-332-0) was classified as Non Corrosive to human skin.

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
GLP compliance:
yes (incl. QA statement)
Test system:
human skin model
Irritation / corrosion parameter:
% tissue viability
Value:
ca. 3.917
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation

The skin irritation classification of Bis(hydroxyethyl)methyloleylammonium chloride was inconclusive.

Test Item

Percentage of viability

(relative to negative control)

Classification

Irritant /non irritant

Bis(hydroxyethyl)methyloleylammonium chloride

3.917 %

Irritant

 

Due to the thick and sticky nature of the test item, the tissues were dosed using an applicator. Unfortunately, at the end of the 60-min exposure time and when removed from the incubator and cooled as per the protocol, the adhesive nature of the test item caused the applicator to stick to the tissue surfaces and upon removal, this lifted the tissues away from the inserts. Additional damage could have been caused when the applicator was removed, therefore, the tissues, only underwent minimal rinsing to prevent further damage and potential tissue loss (submersion in PBS) rather than the full rinsing procedure. As a result, the tissues were potentially exposed to the test item for 42 hrs rather than the 1 hr as described in the SOP. This could therefore, lead to an over prediction of the irritancy (irritant: 3.917% compared to the NC). All acceptance criteria were passed within the scope of the test but because of the deviation caused by the incompatibility of the test item with the dosing method the result is inconclusive.  

 

Table 2:Viability measurements after 60 ± 1 minutes) of application and 42 ± 4 hours post-incubation of test and reference items and controls.

 

Condition

Tissue

Raw data

Blank corrected data

Mean OD

% of Viability

Aliquot 1

Aliquot 2

Aliquot 1

Aliquot 2

NC

Tissue 1

1.792

2.911

1.651

2.770*

1.651

99.970

Tissue 2

1.427

2.418

1.286

2.277

1.781

107.874

Tissue 3

1.503

1.823

1.362

1.682

1.522

92.156

PC

Tissue 1

0.22

0.21

0.079

0.069

0.074

4.452

Tissue 2

0.185

0.189

0.044

0.048

0.046

2.756

Tissue 3

0.178

0.196

0.037

0.055

0.046

2.756

TA1

Tissue 1

0.191

0.177

0.050

0.036

0.043

2.574

Tissue 2

0.212

0.222

0.071

0.081

0.076

4.573

Tissue 3

0.21

0.225

0.069

0.084

0.076

4.603

NC: negative control (DPBS), PC: Positive control (SDS 5%), TA1: Bis(hydroxyethyl)methyloleylammonium chloride. * = removed from the analysis as an outlier and reduced the SD viability to below 18%.

 

Table 3:Mean and SD of cell viability measurements and of viability percentages after 60 min (± 1min) of application and 42h (± 4h) post-incubation.

 

Name

Code

Mean of OD

SD of OD

Mean of viability (%)

SD of viability (%)

CV %

Classification

DPBS

NC

1.651

0.130

100.000

7.859

NI

Non-Irritant

SDS 5%

PC

0.055

0.016

3.321

0.979

I

Irritant

Test Item

TA1

0.065

0.019

3.917

1.163

I

Irritant

 

Prediction model of irritancy: test items that reduce the viability to 50% or below are irritant (I), test items with a percentage viability above 50% are considered to be non-irritant (NI).The percentage of viability obtained with the test itemBis(hydroxyethyl)methyloleylammonium chloridewas 3.917%.

 

Acceptance criteria:

 

Description

Actual values

PASS/FAIL

 

Acceptance criterion 1

 

The mean OD570of the negative control (treated with DPBS) tissues must be ≥ 0.8 and ≤ 2.8

 

1.651

PASS

Acceptance criterion 2

 

The mean of the positive control relative percentage viability must be ≤ 20% of the mean of the negative controls.

 

3.321

PASS

Acceptance criterion 3

 

The standard deviation of viability percentages for triplicate skin models in each experimental condition must be < 18%

 

NC: 7.859

PC: 0.979

TA1: 1.163

PASS

Acceptance criterion 4

 

The mean OD of the 6 wells containing extraction solvent alone (blanks) must be ≤ 0.1.

 

0.142

PASS (Accepted based upon historical system data for blanks; <0.244)

Interpretation of results:
Category 2 (irritant) based on GHS criteria
Conclusions:
Bis (2-Hydroxyethyl) Oleyl Methyl Ammonium Chloride (CAS 18448-65-2, EC 242-332-0) evaluated for skin irritation following OECD Guideline 439 and using EpiDermTM tissue model was found to be irritating.
Executive summary:

The skin irritation potential of Bis(hydroxyethyl)methyloleylammonium chloride was assessed in vitro according to OECD Test Guideline 439: In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method.

 

The percentage of viability obtained with the test item Bis(hydroxyethyl)methyloleylammonium chloride was 3.917%, and therefore according to the prediction model of irritancy Bis(hydroxyethyl)methyloleylammonium chloride is classified as irritating to the skin.

 

However, the results from this study were considered to be inconclusive due to the incompatibility of Bis(hydroxyethyl)methyloleylammonium chloride with the dosing method. During this study, the thick and sticky nature of Bis(hydroxyethyl)methyloleylammonium chloride caused the dosing applicator to adhere to the tissue models and when the tissue applicator was removed from the tissues, there was potential for tissue surface damage and the tissues lifted out of the inserts. Because of this, the tissues were not subjected to the full rinse procedure in case this caused further damage and potential tissue loss. The result was that the tissues were potentially exposed to the test item for 42 hours, instead of the standard 1, which could lead to an over-prediction of the irritancy potential of Bis(hydroxyethyl)methyloleylammonium chloride.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vitro / ex vivo
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
the study does not need to be conducted because the substance is classified as skin irritation and the available information indicates that it should be classified as eye irritation (Category 2)
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification

Categories Display

EU Privacy Disclaimer

This website uses cookies to ensure you get the best experience on our websites.