Registration Dossier

Administrative data

Description of key information

Studies are available that show the substance, C12-14 AO, is irritating to the skin and corrosive to the eyes of rabbits.

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:
2011-09-30 to 2012-01-20
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)
GLP compliance:
yes (incl. certificate)
Test system:
human skin model
Source species:
human
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EST-1000
- Tissue batch number(s): EST-111205-001

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: not reported

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: Single washing step with Dulbecco's phosphate buffered saline (D-PBS). Volume not reported.
- Observable damage in the tissue due to washing: None reported.
- Modifications to validated SOP: None reported

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL
- Incubation time: 3 hours at 37°C
- Spectrophotometer: not specified
- Wavelength: 540 nm

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability/barrier function: Each batch of the epidermal model used meets defined production release criteria, set by the supplier, among which those for viability and for barrier function are the most relevant (MTT, 2 hours Triton X-100: target > 50%). The barrier properties of the tissues were verified by the supplier.
- Morphology: No. of cornified layers - 5; No. of vital cell layers - 4.
- Contamination: The skin model was free of contamination with bacteria (including mycoplasma) or fungi.

NUMBER OF REPLICATE TISSUES: 3

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

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be corrosive to skin if the viability after 3 minutes exposure is less than 50%, or if the viability after 3 minutes exposure is greater than or equal to 50 % and the viability after 1 hour exposure is less than 15%.
- The test substance is considered to be non-corrosive to skin if the viability after 3 minutes exposure is greater than or equal to 50% and the viability after 1 hour exposure is greater than or equal to 15%.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 123.5 μL
- Concentration (if solution): 40.5 %

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 123.5 μL
- Concentration (if solution): 100% (deionised water)

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 50 μL
- Concentration (if solution): 8N
Duration of treatment / exposure:
3 minutes, 1 hour
Duration of post-treatment incubation (if applicable):
3 hours
Number of replicates:
3
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
3 minutes
Value:
78.8
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: mean of 3 replicates
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
1 hour
Value:
65.9
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: mean of 3 replicates
Other effects / acceptance of results:
In comparison to the negative controls, the mean viability of cells exposed to the test item was 78.8% after a 3-minute exposure period and 65.9% after a 1-hour exposure. The OD540 values were well above the cut-off percentage cell viability values distinguishing corrosive from non-corrosive test items of <50% or <15% for a 3-minute or 1-hour treatment, respectively. Therefore, the test item was non-corrosive in this skin model and is predicted to be non-corrosive to human skin.
The mean viability of cells treated with the positive reference item 8 N KOH were 5.0% (3-minute incubation) and 1.2% (1-hour incubation) of the negative controls and were below the cut-off values. Hence, 8 N KOH caused pronounced corrosion in this skin model and is predicted to be corrosive to human skin.
All quality criteria required were fulfilled
Interpretation of results:
GHS criteria not met
Conclusions:
In comparison to the negative controls, the mean viability of cells exposed to the test item was 78.8% after a 3-minute exposure period and 65.9% after a 1-hour exposure. The OD540 values were well above the cut-off percentage cell viability values distinguishing corrosive from non-corrosive test items of <50% or <15% for a 3-minute or 1-hour treatment, respectively. Therefore, the test item was non-corrosive in this skin model and is predicted to be non-corrosive to human skin.
Executive summary:

In an in vitro skin corrosivity study performed in accordance with OECD Guideline 431 using the EST-100 model the test item, N,N-dimethyldecylamine-N-oxide (solution), was applied to the skin surface. De-ionised water was used as the negative control. 8 N KOH was used as the positive reference item. Two exposure times of 3 minutes or 1 hour were employed.

In comparison to the negative controls, the mean viability of cells exposed to the test item was 78.8% after a 3-minute exposure period and 65.9% after a 1-hour exposure. The OD540values were well above the cut-off percentage cell viability values distinguishing corrosive from non-corrosive test items of <50% or<15% for a 3-minute or 1-hour treatment, respectively. Therefore, the test item was non-corrosive in this skin model and is predicted to be non-corrosive to human skin.

The mean viability of cells treated with the positive reference item 8 N KOH were 5.0% (3-minute incubation) and 1.2% (1-hour incubation) of the negative controls and were below the cut-off values. Hence, 8 N KOH caused pronounced corrosion in this skin model and is predicted to be corrosive to human skin.

Under the present test conditions N,N-dimethyldecylamine-N-oxide (solution) tested at two exposure times of 3 minutes or 1 hour was non-corrosive to skin in vitro.

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2011-09-28 to 2012-01-20
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
other: OECD Guideline No. 439
Qualifier:
according to
Guideline:
other: EC method B46
GLP compliance:
yes (incl. certificate)
Test system:
human skin model
Source species:
human
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EST-1000
- Tissue batch number(s): EST-111205-001

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 21°C
- Temperature of post-treatment incubation (if applicable): not reported - 42 hours incubation

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: Single washing step with Dulbecco's phosphate buffered saline (D-PBS). Volume not reported.
- Observable damage in the tissue due to washing: None reported
- Modifications to validated SOP: None reported

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL
- Incubation time: 3 hours at 37°C
- Spectrophotometer: not specified
- Wavelength: 540 nm

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability/Barrier function: Each batch of the epidermal model used meets defined production release criteria, set by the supplier, among which those for viability and for barrier function are the most relevant (MTT, 2 hours Triton X-100: target > 50%). The barrier properties of the tissues were verified by the supplier.
- Morphology: No. of cornified layers - 5; No. of vital cell layers - 4.
- Contamination: The skin model was free of contamination with bacteria (including mycoplasma) or fungi.

NUMBER OF REPLICATE TISSUES: 3

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

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be irritant to skin if the viability after exposure and post-treatment incubation is less than 50%
- The test substance is considered to have no category if the viability ater exposure and post-treatment incubation is greater than 50%
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 74.1 μL
- Concentration (if solution): 40.5%

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 74.1 μL
- Concentration (if solution): 100% (deionised water)

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 30 μL
- Concentration (if solution): 5%
Duration of treatment / exposure:
20 minutes
Duration of post-treatment incubation (if applicable):
42 hours
Number of replicates:
3
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
20 minutes exposure
Value:
74
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: mean of 3 replicates
Other effects / acceptance of results:
The mean viability of the cells exposed to the test item was 74.0% of the mean negative control value. The OD540 values were well above the cut-off percentage cell viability value that distinguishes irritant from non-irritant test items of >50% for a 20-minute exposure.
The test item was considered to be non-cytotoxic and predicted to be not irritant to skin.
The viability of cells treated with the positive reference item 5% SDS, was 8.3% of the negative controls and was below the cut-off value. Hence, 5% SDS is predicted to cause pronounced skin irritation.
All quality criteria required were fulfilled.
Interpretation of results:
GHS criteria not met
Conclusions:
The mean viability of the cells exposed to the test item was 74.0% of the mean negative control value. The OD540 values were well above the cut-off percentage cell viability value that distinguishes irritant from non-irritant test items of >50% for a 20-minute exposure.
The test item was considered to be non-cytotoxic and predicted to be not irritant to skin.
Executive summary:

In an in vitro skin irritation study performed in accordance with OECD Guideline 439 and EC method B46, using the EST-100 model the test item, N,N-dimethyldecylamine-N-oxide (solution), was applied to the model skin surface. De-ionised water was used as the negative control. 5% aqueous sodium dodecyl sulphate (SDS) was used as the positive reference item. The cell viability was measured by determining the optical density (OD) at a wavelength of 540 nm. An exposure time of 20 minutes was employed.

The mean viability of cells exposed to the test item was 74.0% of the negative controls. The mean OD540value was well above the cut-off percentage cell viability value of > 50% that distinguishes irritant from non-irritant test items after a 20-minute exposure. The test item was considered to be non-cytotoxic and is predicted to be not irritant to skin.

The viability of cells treated with the positive reference item, 5% SDS, was 8.3% of the negative controls and below the 50% cut-off value. Hence, 5% SDS is predicted to cause pronounced skin irritation.

Under the present test conditions N,N-dimethyldecylamine-N-oxide (solution), tested at an exposure time of 20 minutes, was non-cytotoxic and not irritant to skin in vitro.

 

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
24 May 2012-01 August 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
Himalayan
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: LPT Laboratory of Pharmacology and Toxicology GmbH & Co. KG, Branch Löhndorf, 24601 Löhndorf/Post Wankendorf, Germany
- Age at study initiation:4.5-6.5 months
- Weight at study initiation: 2.6-3.0 kg
- Housing: The animals were kept singly in cages measuring 380 mm x 425 mm x 600 mm (manufacturer: Dipl. Ing. W. EHRET GmbH, 16352 Schön¬walde, Germany)
- Diet: Commercial diet, ssniff® K-H V2333 (ssniff Spezialdiäten GmbH, 59494 Soest, Germany, served as food. The food was available ad libitum before and after the exposure period.
- Water: Drinking water was offered ad libitum before and after the exposure period.
- Acclimation period: At least 20 adaptation days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): No information
- Photoperiod (hrs dark / hrs light): 12h/12h

IN-LIFE DATES: From: To: 01 June - 19 June, 2012
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): 40.5 % w/w

Duration of treatment / exposure:
4 hours
Observation period:
7 days
Number of animals:
3
Details on study design:
TEST SITE
- Area of exposure: approx 6 cm2
- Type of wrap if used: Gauze

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Not applicable
- Time after start of exposure: Not applicable

SCORING SYSTEM: Draize
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 4 days
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 4 days
Irritation parameter:
edema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
edema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
edema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritant / corrosive response data:
Time after removal of patch Animal #1 Animal #2 Animal #3
Erythema/Oedema Erythema/Oedema Erythema/Oedema
60 min 1/0 1/0 1/0
24 h 1/0 1/0 1/0
48 h 1/0 1/0 1/0
72 h 1/0 1/0 1/0
4 d 0/0 1/0 0/0
5 d -/- 1/0 -/-
6 d -/- 1/0 -/-
7 d -/- -/- -/-
Interpretation of results:
GHS criteria not met
Conclusions:
Based on the results of this study, C10 AO is not corrosive or irritant to the skin of rabbits.
Executive summary:

Under the present test conditions, three rabbits exposed for 4 hours to 0.5 mL N,N-dimethyldecylamine-N-oxide/patch (semi-occlusive conditions) showed the following effect:

Erythema (grade 1) was observed in all animals 60 minutes to 72 hours after patch removal, and in animal no. 2 until 6 days after patch removal.

According to EC Regulation 1272/2008 and subsequent regulations, the test item is non-irritating and no labelling is required.

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
No data
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
according to
Guideline:
other: No data
Deviations:
not applicable
Principles of method if other than guideline:
The test material was evaluated for skin irritation to normal and abraded skin of eight New Zealand white rabbits as a 2 % aqueous solution. Observations were recorded at 24- and 72 h.
GLP compliance:
no
Remarks:
Study pre-dates GLP.
Species:
rabbit
Strain:
New Zealand White
Type of coverage:
occlusive
Preparation of test site:
other: Abraded and non-abraded.
Vehicle:
water
Controls:
not specified
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL
- Concentration (if solution): 0.6 % AO in water

Duration of treatment / exposure:
24 h
Observation period:
72 h
Number of animals:
Eight
Details on study design:
TEST SITE
- Area of exposure: 1.5x2 inches
- % coverage: no data
- Type of wrap if used: test material applied to a plastic coverlet, secured in place using plastic tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no data


SCORING SYSTEM: Draize
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: Mean of 24 & 72 hrs
Score:
0.5
Max. score:
4
Reversibility:
other: 1/8 animals had a score of 1 at 72 hrs
Remarks on result:
other: maximum score observed at 72 hrs = 1
Irritation parameter:
edema score
Basis:
mean
Time point:
other: Mean of 24 & 72 hrs
Score:
0.125
Max. score:
4
Reversibility:
fully reversible
Remarks on result:
other: Maximum score at 24 h = 1
Other effects:
Throughout the treatment and observation period all animals appeared clinically normal. No evidence of pain was observed. All animals thrived and responded normally to external stimuli.
Interpretation of results:
GHS criteria not met
Conclusions:
A 2 % solution of the test material caused slight erythema and occasional oedema on the abraded and non-abraded skin over the 72 h observation period. The 2 % solution of the test material (equivalent to 0.6 % AO) showed a primary irritation index of 0.93.
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
24-05-1982 to 26-05-1982
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
according to
Guideline:
other: U.S. Federal Register, Vol. 41 (188): 42572 (27th September 1976). US CFR 49(173).
Deviations:
not specified
GLP compliance:
no
Remarks:
Study pre-dates GLP.
Species:
rabbit
Strain:
New Zealand White
Type of coverage:
occlusive
Preparation of test site:
other: Clipped
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): 30 %w/w
Duration of treatment / exposure:
4 h
Observation period:
48 h
Number of animals:
Six
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 4 h
Score:
0.33
Max. score:
4
Reversibility:
no data
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 4 h
Score:
0
Max. score:
4
Reversibility:
other: Not applicable
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 48 h
Score:
2.33
Max. score:
4
Reversibility:
no data
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 48 h
Score:
0.66
Max. score:
4
Reversibility:
no data
Other effects:
No data
Interpretation of results:
other: Not corrosive
Remarks:
Criteria used for interpretation of results: other: US Federal Register.
Conclusions:
The study indicates that the test substance does not have a corrosive action on the skin following a 4 h exposure period.
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
10-1976
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
according to
Guideline:
other: US Federal Register Vol. 38 (187): 1500:41, 1973.
Deviations:
not specified
GLP compliance:
no
Remarks:
Study pre-dates GLP.
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: stated as 'recognised breeders'
- Age at study initiation: 12-14 weeks
- Weight at study initiation: average bw 2.15 kg
- Housing: caged singly
- Diet: Rank Hovis MacDougal rabbit diet, ad libitum
- Water: filtered water ad libitum
- Acclimation period:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 +/- 2
- Humidity (%): 50-70
- Air changes (per hr): not stated
- Photoperiod: natural lighting conditions

Type of coverage:
occlusive
Preparation of test site:
other: intact and abraded
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): 6.5 % as supplied
Duration of treatment / exposure:
24 hours
Observation period:
72 h
Number of animals:
Six
Details on study design:
TEST SITE
- Area of exposure: 2.5 x 2.5 cm
- % coverage: 10 %
- Type of wrap if used: Smith & Nephew Elastic bandage (occlusive).

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Not done
- Time after start of exposure:

SCORING SYSTEM: Draize
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
other: mean of 24 & 72 hours
Score:
0.5
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
other: intact skin
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
other: mean of 24 & 72 hours
Score:
1.5
Max. score:
4
Reversibility:
not fully reversible within: 72 hours
Remarks on result:
other: intact skin
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
other: mean of 24 & 72 hours
Score:
1.5
Max. score:
4
Reversibility:
not fully reversible within: 72 hours
Remarks on result:
other: intact skin
Irritation parameter:
erythema score
Basis:
animal #4
Time point:
other: mean of 24 & 72 hours
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
other: intact skin
Irritation parameter:
erythema score
Basis:
animal #5
Time point:
other: mean of 24 & 72 hours
Score:
1.5
Max. score:
4
Reversibility:
not fully reversible within: 72 hours
Remarks on result:
other: intact skin
Irritation parameter:
erythema score
Basis:
animal #6
Time point:
other: mean of 24 & 72 hours
Score:
1.5
Max. score:
4
Reversibility:
not fully reversible within: 72 hours
Remarks on result:
other: intact skin
Irritation parameter:
edema score
Basis:
animal #1
Time point:
other: mean of 24 & 72 hours
Score:
0.5
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
other: intact skin
Irritation parameter:
edema score
Basis:
animal #2
Time point:
other: mean of 24 & 72 hours
Score:
0.5
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
other: intact skin
Irritation parameter:
edema score
Basis:
animal #3
Time point:
other: mean of 24 & 72 hours
Score:
0.5
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
other: intact skin
Irritation parameter:
edema score
Basis:
animal #4
Time point:
other: mean of 24 & 72 hours
Score:
0.5
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
other: intact skin
Irritation parameter:
edema score
Basis:
animal #5
Time point:
other: mean of 25 & 72 hours
Score:
1
Max. score:
4
Reversibility:
not fully reversible within: 72 hours
Remarks on result:
other: intact skin
Irritation parameter:
edema score
Basis:
animal #6
Time point:
other: mean of 24 & 72 hours
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
other: intact skin
Other effects:
No data
Interpretation of results:
GHS criteria not met
Conclusions:
The test substance gave a primary irritation score of 2.07 over 72 h which is classed as a moderate irritant to the skin of the rabbit according to the author of the study report. Based on EU classification criteria, the test substance is not classified as irritating to skin.
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
01-06-1981 to 03-06-1981
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
according to
Guideline:
other: US Federal Register Vol. 41 (188): 42572 (27th September 1976). US CFR 49 (173).
Deviations:
not specified
GLP compliance:
no
Remarks:
Study pre-dates GLP.
Species:
rabbit
Strain:
New Zealand White
Type of coverage:
occlusive
Preparation of test site:
other: Clipped
Vehicle:
unchanged (no vehicle)
Controls:
not specified
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL
- Concentration (if solution): 30 %w/w

Duration of treatment / exposure:
4 h
Observation period:
48 h
Number of animals:
6
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 48 h
Score:
2
Max. score:
4
Reversibility:
no data
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 48 h
Score:
0.66
Max. score:
4
Reversibility:
no data
Other effects:
No data
Interpretation of results:
other: Not Corrosive
Remarks:
Criteria used for interpretation of results: other: US Federal Register
Conclusions:
No reactions were apparent at the 4 h observation. Well-defined erythema was apparent at all 6 treated sites at the 48 h observation and very slight oedema was apparent at 4 treated sites at this time. The test substance did not cause destruction of intact skin of the albino rabbit and based on this result would not be regarded as a corrosive agent according to the Definitions in the US Federal Register Test. The Class 8 classification of the UN Economics and Social Council was slightly corrosive.
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1978-02-22
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
yes
Remarks:
- Treatment time is 24 hours instead of 4 hours. 0.4ml of test substance dosed instead of 0.5ml. Systemic toxicity, body weight and histopathology results are not given.
Principles of method if other than guideline:
None
GLP compliance:
no
Remarks:
pre-GLP
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Kuiper's rabbit ranch, Gary, Indiana
- Age at study initiation: Not available
- Weight at study initiation: 2103 - 2781 gm
- Housing: Individually in hanging wire-mesh cages in temperature and humidity controlled quarters
- Diet (e.g. ad libitum): Purina rabbit chow, ad libitum
- Water (e.g. ad libitum): Water, ad libitum
- Acclimation period: 16 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): As per S.O.P.
- Humidity (%): As per S.O.P.
- Air changes (per hr): As per S.O.P.
- Photoperiod (hrs dark / hrs light): As per S.O.P.


IN-LIFE DATES: 1978-02-22
Type of coverage:
occlusive
Preparation of test site:
abraded
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.4 ml of test substance
- Concentration (if solution): Undiluted


VEHICLE
- Amount(s) applied (volume or weight with unit): Not applicable
- Concentration (if solution): Not applicable
- Lot/batch no. (if required): Not applicable
- Purity: Not applicable
Duration of treatment / exposure:
0.4 ml of undiluted P7270 was applied to one abraded and one intact test site on each of the three New Zealand white rabbits (2 male and 1 female)
using 38x38 mmParke Davis Readi-bandage with a 20x20 mm felt pad secured with wrappings of 75 mm elastoplast tape for 24 hours. After 24 hours the test sites were wiped with a damp cloth. The sites were examined for erythema and edema using Draize method of scoring at 24 and 72 hours.
Observation period:
24 and 72 hours
Number of animals:
3 animals (2 male and 1 female)
Details on study design:
TEST SITE
- Area of exposure: Back of animals (intact and abraded skin site)
- % coverage: Test substance applied using 38x38 mmParke Davis Readi-bandage with a 20x20 mm felt pad
- Type of wrap if used: 75 mm elastoplast tape


REMOVAL OF TEST SUBSTANCE
- Washing (if done): Test substance removed with a damp cloth
- Time after start of exposure: 24 hours


SCORING SYSTEM:
Erythema and eschar scoring from 0-4 with 0 = no erythema, 1 = very slight erythema (barely perceptible), 2 = well defined erythema, 3 = moderate to severe erythema, 4 = severe erythema (beet redness) to slight eschar formation (injuries in depth).
Edema scoring from 0-4 with 0 = no edema, 1 = very slight edema (barely perceptible), 2 = slight edema (edges of area well defined by definite raising), 3 = moderate edema (raised approximately 1mm), 4 = severe edema (raised more than 1 mm nd extending beyond the area of exposure).
Primary irritation index: 0-2 = mild, 2-5 = moderately irritating, 5-6 = moderately to severely irritating, 6-8 = severely irritating
Irritation parameter:
primary dermal irritation index (PDII)
Basis:
mean
Time point:
other: 24 hours
Score:
2
Max. score:
8
Reversibility:
not reversible
Remarks:
in 72 hrs
Remarks on result:
other: Intact and abraded skin site
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 24 hours
Score:
1
Max. score:
4
Reversibility:
not reversible
Remarks:
in 72 hours
Remarks on result:
other: Intact and abraded skin site
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 24 hours
Score:
1
Max. score:
4
Reversibility:
not reversible
Remarks:
in 72 hours
Remarks on result:
other: Intact and abraded skin site
Irritation parameter:
primary dermal irritation index (PDII)
Basis:
mean
Time point:
other: 72 hours
Score:
8
Max. score:
8
Reversibility:
not reversible
Remarks:
in 72 hours
Remarks on result:
other: Intact and abraded skin site
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 72 hours
Score:
4
Max. score:
4
Reversibility:
not reversible
Remarks:
in 72 hours
Remarks on result:
other: Intact and abraded skin site
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 72 hours
Score:
4
Max. score:
4
Reversibility:
not reversible
Remarks:
in 72 hours
Remarks on result:
other: Intact and abraded skin site
Irritant / corrosive response data:
For Intact and Abraded skin site- PII at 24 hours: 2.0, PII at 72 hours: 8.0
Other effects:
One animal showed blanching at 24 hours at abraded skin site.

 Animal No.

 Skin site

Degree of erythema after---- [Observation time]

 Degree of edema after---- [Observation time]  

 Ø 24/72 h >= 2.3 but < 4.0   

 

 

 24 h

 72 h

 24 h

 72 h

 Erythema

 Edema

 

 Intact

 

 

 

 

 

 

 30970

 

 1

 4

 1

 4

Yes 

 Yes  

 

 

Ø 24/ 72 h = 2.5

 Ø 24/ 72 h = 2.5

  => Positive responder

 30979

 

 1

 4

 1

 4

  Yes

  Yes

 

 

 Ø 24/ 72 h = 2.5

 Ø 24/ 72 h = 2.5

  => Positive responder

 31017

 

 1

 4

 1

 4

  Yes

  Yes

 

 

Ø 24/ 72 h = 2.5

Ø 24/ 72 h = 2.5     

  => Positive responder

 

 Abraded

 

 

 

 

 

 

 30970

 

 1

 4

 1

 4

  Yes

  Yes

 

 

  Ø 24/ 72 h = 2.5

  Ø 24/ 72 h = 2.5

  => Positive responder

 30979

 

 1

 4

 1

 4

  Yes

  Yes

 

 

  Ø 24/ 72 h = 2.5

  Ø 24/ 72 h = 2.5

  => Positive responder

31017

 

 1

 4

 1

 4

  Yes

  Yes

 

 

  Ø 24/ 72 h = 2.5

  Ø 24/ 72 h = 2.5  

=> Positive responder    

Interpretation of results:
Category 2 (irritant) based on GHS criteria
Conclusions:
P7270 (28% DDAO) is classified as a Category 2 skin irritant according to EU GHS classification.
Executive summary:

0.4 ml of undiluted P7270 (28% DDAO) was applied to one abraded and one intact test site on each of the three New Zealand white rabbits (2 male and 1 female) using 38x38 mmParke Davis Readi-bandage with a 20x20 mm felt pad secured with wrappings of 75 mm elastoplast tape for 24 hours. After 24 hours the test sites were wiped with a damp cloth. The sites were examined for erythema and edema using Draize method of scoring at 24 and 72 hours.

Occlusive 24 hour exposure to 0.4 ml of undiluted DDAO produced very slight erythema and very slight edema at the intact and abraded sites of all the test animals at 24 hours. At 72 hours severe erythema (beet redness) to slight eschar formation (injuries in depth) and severe edema (raised more than 1 mm extending beyond the area of exposure) was served at the intact and abraded of all the test animals.

One animal showed blanching at 24 hours at abraded skin site.

Thus, 28% DDAO/ C10-C16 alkyldimethyl, N-Oxide is classified as a Category 2 skin irritant according to EU GHS classification.

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1978-09-21
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
yes
Remarks:
- Treatment time is 24 hours instead of 4 hours. 0.4ml of test substance dosed instead of 0.5ml. Systemic toxicity, body weight and histopathology results are not given.
Principles of method if other than guideline:
None
GLP compliance:
no
Remarks:
pre-GLP
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Kings wheel
- Age at study initiation: Not available
- Weight at study initiation: 2058.9 - 3285.9 gm
- Housing: Individually in stainless steel cages with stainless steel slatted flooring
- Diet (e.g. ad libitum): Purina lab rabbit chow
- Water (e.g. ad libitum): Tap water; ad libitum
- Acclimation period: Minimum 7 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.4°C - 21.1°C
- Humidity (%): 72%
- Air changes (per hr): Not available
- Photoperiod (hrs dark / hrs light): 12 hours on/off fluorescent lighting


IN-LIFE DATES: 1978-09-21
Type of coverage:
occlusive
Preparation of test site:
abraded
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.4 ml of P0434
- Concentration (if solution): Undiluted


VEHICLE
- Amount(s) applied (volume or weight with unit): Not applicable
- Concentration (if solution): Not applicable
- Lot/batch no. (if required): Not applicable
- Purity: Not applicable
Duration of treatment / exposure:
0.4 ml of undiluted P0434 was applied to one abraded and one intact test site on each of the three female New Zealand white rabbits
using Parke Davis Readi-bandage secured with hypo-allergenic tape. Test sites were wrapped with gauze and elastoplast tape for 24 hours. After 24 hours the test sites were wiped with a wet paper towel. The sites were examined for erythema and edema using Draize method of scoring at 24 and
72 hours.
Observation period:
24 and 72 hours
Number of animals:
3 female rabbits
Details on study design:
TEST SITE
- Area of exposure: Back of animals (right side - abraded, left side - intact)
- % coverage: Test substance applied using Parke-Davis Readi-Bandage
- Type of wrap if used: Gauze and elastoplast tape


REMOVAL OF TEST SUBSTANCE
- Washing (if done): Test sites wiped with wet paper towel
- Time after start of exposure: 24 hours


SCORING SYSTEM: Draize method
Erythema and eschar scoring from 0-4 with 0 = no erythema, 1 = very slight erythema (barely perceptible), 2 = well defined erythema, 3 = moderate to severe erythema, 4 = severe erythema (beet redness) to slight eschar formation (injuries in depth).
Edema scoring from 0-4 with 0 = no edema, 1 = very slight edema (barely perceptible), 2 = slight edema (edges of area well defined by definite raising), 3 = moderate edema (raised approximately 1mm), 4 = severe edema (raised more than 1 mm nd extending beyond the area of exposure).
Primary irritation index: 0-2 = mild, 2-5 = moderately irritating, 5-6 = moderately to severely irritating, 6-8 = severely irritating
Irritation parameter:
edema score
Basis:
animal #1
Time point:
24 h
Score:
0
Max. score:
4
Reversibility:
other: no edema was observed
Remarks on result:
other: intact skin site
Irritation parameter:
edema score
Basis:
animal #1
Time point:
48 h
Remarks on result:
not measured/tested
Remarks:
intact skin site
Irritation parameter:
edema score
Basis:
animal #1
Time point:
72 h
Score:
0
Max. score:
4
Reversibility:
other: no edema was observed
Remarks on result:
other: intact skin site
Irritation parameter:
edema score
Basis:
animal #2
Time point:
24 h
Score:
0
Max. score:
4
Reversibility:
other: no edema was observed
Remarks on result:
other: intact skin site
Irritation parameter:
edema score
Basis:
animal #2
Time point:
48 h
Remarks on result:
not measured/tested
Remarks:
intact skin site
Irritation parameter:
edema score
Basis:
animal #2
Time point:
72 h
Score:
0
Max. score:
4
Reversibility:
other: no edema was observed
Remarks on result:
other: intact skin site
Irritation parameter:
edema score
Basis:
animal #3
Time point:
24 h
Score:
0
Max. score:
4
Reversibility:
other: no edema was observed
Remarks on result:
other: intact skin site
Irritation parameter:
edema score
Basis:
animal #3
Time point:
48 h
Remarks on result:
not measured/tested
Remarks:
intact skin site
Irritation parameter:
edema score
Basis:
animal #3
Time point:
72 h
Score:
0
Max. score:
4
Reversibility:
other: no edema was observed
Remarks on result:
other: intact skin site
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24 h
Score:
1
Max. score:
4
Reversibility:
not reversible
Remarks:
in 72 hours
Remarks on result:
other: intact skin site
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
48 h
Remarks on result:
not measured/tested
Remarks:
intact skin site
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
72 h
Score:
4
Max. score:
4
Reversibility:
not reversible
Remarks:
in 72 hours
Remarks on result:
other:
Remarks:
intact skin site
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
24 h
Score:
2
Max. score:
4
Reversibility:
not reversible
Remarks:
in 72 hours
Remarks on result:
other: intact skin site
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
48 h
Remarks on result:
not measured/tested
Remarks:
intact skin site
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
72 h
Score:
4
Max. score:
4
Reversibility:
not reversible
Remarks:
in 72 hours
Remarks on result:
other: intact skin site
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
24 h
Score:
2
Max. score:
4
Reversibility:
not reversible
Remarks:
in 72 hours
Remarks on result:
other: intact skin site
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
48 h
Remarks on result:
not measured/tested
Remarks:
intact skin site
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
72 h
Score:
4
Max. score:
4
Reversibility:
not reversible
Remarks:
in 72 hours
Remarks on result:
other: intact skin site
Irritant / corrosive response data:
For intact and abraded skin site at 24 hours PII is 1.67 and at 72 hours PII is 4.0.
Other effects:
None reported

 Animal No.

 Skin site

Degree of erythema after---- [Observation time]

 Degree of edema after---- [Observation time]  

 Ø 24/72 h >= 2.3 but < 4.0   

 

 

 24 h

 72 h

 24 h

 72 h

 Erythema

 Edema

 

 Intact

 

 

 

 

 

 

 RB 608 78

 

 1

 4

 0

 0

Yes 

 No  

 

 

Ø 24/ 72 h = 2.5

 Ø 24/ 72 h = 0

  => Positive responder

 RB 524 78

 

 2

 4

 0

 0

  Yes

  No

 

 

 Ø 24/ 72 h = 3.0

 Ø 24/ 72 h = 0

  => Positive responder

 RB 525 78

 

 2

 4

 0

 0

  Yes

  No

 

 

Ø 24/ 72 h = 3.0

Ø 24/ 72 h = 0     

  => Positive responder

 

 Abraded

 

 

 

 

 

 

 RB 608 78

 

 1

 4

 0

 0

  Yes

  No

 

 

  Ø 24/ 72 h = 2.5

  Ø 24/ 72 h = 0

  => Positive responder

 RB 524 78

 

 2

 4

 0

 0

  Yes

  No

 

 

  Ø 24/ 72 h = 3.0

  Ø 24/ 72 h = 0

  => Positive responder

RB 525 78

 

 2

 4

 0

 0

  Yes

  No

 

 

  Ø 24/ 72 h = 3.0

  Ø 24/ 72 h = 0  

=> Positive responder    

Interpretation of results:
Category 2 (irritant) based on GHS criteria
Conclusions:
P0434 (27.8% DDAO) is classified as a Category 2 skin irritant according to EU GHS classification.
Executive summary:

0.4 ml of undiluted P0434 (27.8% DDAO) was applied to one abraded and one intact test site on each of the three female New Zealand white rabbits using Parke Davis Readi-bandage secured with hypo-allergenic tape. Test sites were wrapped with gauze and elastoplast tape for 24 hours. After 24 hours the test sites were wiped with a wet paper towel. The sites were examined for erythema and edema using Draize method of scoring at 24 and 72 hours.

Very slight to well-defined erythema at the intact and abraded sites of all the test animals at 24 hours. At 72 hours severe erythema (beet redness) to slight eschar formation (injuries in depth) were observed at the intact and abraded of all the test animals. No edema was observed at any time point.

Thus, 27.8% DDAO/ C10-C16 alkyldimethyl, N-Oxide is classified as a Category 2 skin irritant according to EU GHS classification.

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to
Guideline:
other: FHSA/CPSC Design, 16 CFR 1500
GLP compliance:
no
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Kuiper Rabbitry, Gary, Indiana, USA
- Age at study initiation: 8-10 weeks old
- Weight at study initiation: no data
- Housing: individually housed in stainless steel cages
- Diet: Purina rabbit chow ad libitum
- Water: ad libitum
- Acclimation period: minimum 4 days

ENVIRONMENTAL CONDITIONS
Temperature, humidy and light were controlled according to the receommendations contained in DHEW Publication No. 86-23 (NIH) Guide for the Care and Use of Laboratory Animals
Type of coverage:
occlusive
Preparation of test site:
other: shaved and shaved + abraded
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied: 0.5 mL
- Concentration (if solution): 5 % active or 2.5 % active in aqueous solution as supplied

Duration of treatment / exposure:
4 hours
Observation period:
24 and 72 hours
Number of animals:
3 animals per dose level
Details on study design:
TEST SITE
- Area of exposure: 2.5 cm2
- % coverage: not stated
- Type of wrap if used:Site was covered with a two layer gauze patch secured with non-irritating porous adhesive tape and the entire site was wrapped with a 4 mil plastic wrap and secured with more tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Excess material was removed from the site (method not specified)
- Time after start of exposure: 4 hours

SCORING SYSTEM: Draize
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
other: average of 24 & 72 hours scores
Score:
0.5
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
other: 5 % AO solution
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
other: average of 24 & 72 hour scores
Score:
0
Max. score:
4
Remarks on result:
other: 5 % AO solution
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
other: average of 24 & 72 hour scores
Score:
1.5
Max. score:
4
Reversibility:
not fully reversible within: 72 hours
Remarks on result:
other: 5 % AO solution
Irritation parameter:
edema score
Basis:
animal #1
Time point:
other: average of 24 & 72 hour scores
Score:
0
Max. score:
4
Remarks on result:
other: 5 % AO solution
Irritation parameter:
edema score
Basis:
animal #2
Time point:
other: average of 24 & 72 hour scores
Score:
0
Max. score:
4
Remarks on result:
other: 5 % AO solution
Irritation parameter:
edema score
Basis:
animal #3
Time point:
other: average of 24 & 72 hour scores
Score:
0
Max. score:
4
Remarks on result:
other: 5 % AO solution
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
other: average of 24 & 27 hour scores
Score:
0.5
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
other: 2.5 % AO Solution
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
other: average of 24 & 72 hour scores
Score:
0
Max. score:
4
Remarks on result:
other: 2.5 % AO Solution
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
other: average of 24 & 72 hour scores
Score:
0
Max. score:
4
Remarks on result:
other: 2.5 % AO Solution
Irritation parameter:
edema score
Basis:
animal #1
Time point:
other: average of 24 & 72 hour scores
Score:
0
Max. score:
4
Remarks on result:
other: 2.5 % AO Solution
Irritation parameter:
edema score
Basis:
animal #2
Time point:
other: average of 24 & 72 hour scores
Score:
0
Max. score:
4
Remarks on result:
other: 2.5 % AO Solution
Irritation parameter:
edema score
Basis:
animal #3
Time point:
other: average of 24 & 72 hour scores
Score:
0
Max. score:
4
Remarks on result:
other: 2.5 % AO Solution

Only the results from intact skin are used in the scoring

Interpretation of results:
GHS criteria not met
Conclusions:
In this study solutions containing C12-14 amine oxide at 2.5 or 5 %w/w were not irritating to the skin of rabbits.
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Hoechst AG, Kastengrund, conventional breeding
- Age at study initiation: no data
- Weight at study initiation: 2.6-3.4 kg
- Housing:no data
- Diet (e.g. ad libitum): Altromin 2123 Haltungsdiat ad libitum
- Water (e.g. ad libitum): deionised, chlorinated water ad libitum
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/-2
- Humidity (%): 55 +/- 10
- Air changes (per hr): air conditioned room, no data
- Photoperiod (hrs dark / hrs light): 12 hours light/dark

Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
unchanged (no vehicle)
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL
- Concentration (if solution): 30 % w/v

Duration of treatment / exposure:
4 hours
Observation period:
14 days
Number of animals:
3
Details on study design:
TEST SITE
- Area of exposure: 2.5x2.5 cm
- % coverage:
- Type of wrap if used: semi-occlusive binding, no further details

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no description of method
- Time after start of exposure: 4 hours

SCORING SYSTEM: Draize
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2.7
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritation parameter:
edema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Irritation parameter:
edema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0.6
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
edema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Interpretation of results:
Category 2 (irritant) based on GHS criteria
Conclusions:
The C12-18 AO is a skin irritant in this study.
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
24-05-1982 to 26-05-1982
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
according to
Guideline:
other: US Federal Register Vol. 41 (188): 24572 (27th September 1976). US CFR 49 (173).
Deviations:
not specified
GLP compliance:
no
Remarks:
Study pre-dates GLP.
Species:
rabbit
Strain:
New Zealand White
Type of coverage:
occlusive
Preparation of test site:
other: Clipped
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): 30 %w/w
Duration of treatment / exposure:
4 h
Observation period:
48 h
Number of animals:
6 animals
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 4 h
Score:
0.66
Max. score:
4
Reversibility:
not fully reversible within: the 48 h observation period.
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 4 h
Score:
0
Max. score:
4
Reversibility:
other: Not applicable
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 48 h
Score:
1.83
Max. score:
4
Reversibility:
not fully reversible within: the 48 h observation period
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 48 h
Score:
0
Max. score:
4
Reversibility:
other: Not applicable
Other effects:
No data
Interpretation of results:
other: Not corrosive
Remarks:
Criteria used for interpretation of results: other: US Federal Register
Conclusions:
Slight erythema was apparent at 4 of the 6 treated sites following a 4 h exposure to the test material. Slight increase in the observable reactions was apparent during the course of the study. No necrosis or other evidence of corrosive action was apparent during the study. The results indicate that the test material does not have a corrisive action on the skin following a 4 h exposure period.
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP study to US EPA guideline. Method deviates from current guideline but provides sufficient detail.
Qualifier:
according to
Guideline:
other: US Federal Register, Vol 41, part 188 p42572 1976
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Recognised breeders. Premises inspected by testing facility
- Age at study initiation: Not specified
- Weight at study initiation: Not specified
- Housing: Grid bottomed galvanised metal cages.
- Diet (e.g. ad libitum): 90 - 95g pelleted commercially available rabbit food per day
- Water (e.g. ad libitum): ad libitum
- Acclimation period: Not specified. Rabbits placed in restrains and shaved at least one hour before treatment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): air-conditioned
- Humidity (%): Not specified
- Air changes (per hr): Not specified
- Photoperiod (hrs dark / hrs light): 14h light/10h dark

IN-LIFE DATES: From: 1982-05-24 To: 1982-05-26
Type of coverage:
occlusive
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


Duration of treatment / exposure:
4h
Observation period:
48 h
Number of animals:
6 female
Details on study design:
TEST SITE
- Area of exposure: 2.5 cm
- % coverage: Not specified
- Type of wrap if used: "Sleek" plastic adhesive wrapping over lint pad. Elastic adhesive bandage (7.5 cm wide) used to secure lint pad.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): tissue soaked in sterile distilled water
- Time after start of exposure: 4 h

SCORING SYSTEM: Draize
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 4 h
Score:
0.66
Max. score:
4
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 48 h
Score:
2.83
Max. score:
4
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 4 h
Score:
0
Max. score:
4
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 48 h
Score:
1.17
Max. score:
4

Rabbit number

4h observation

48h observation

Erythema

Oedema

Erythema

Oedema

1

1

0

3

2

2

1

0

2

1

3

0

0

3

0

4

1

0

3

1

5

0

0

3

1

6

1

0

3

2

Total

4

0

17

7

Interpretation of results:
other: Not corrosive
Remarks:
Criteria used for interpretation of results: other: UN Classification (TRANS/GE.15/R.275, Annex 3, page 1, Revision of Class 8 of RID and ADR, October 1977)
Conclusions:
The substance is not classified as corrosive under the conditions of the study.
Executive summary:

In a GLP skin corrosivity study performed to US Federal Guidelines the substance tested as provided was not considered to be corrosive to the skin of rabbits in accordance with UN classification criteria for corrosivity.

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
according to
Guideline:
other: CFR 16.1500.41
GLP compliance:
not specified
Species:
rabbit
Type of coverage:
not specified
Preparation of test site:
other: abraded and intact
Vehicle:
water
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL
- Concentration (if solution): 5% active
Duration of treatment / exposure:
72 hours
Number of animals:
6
Irritation parameter:
erythema score
Basis:
mean
Remarks:
of six animals
Time point:
other: mean of 24 & 72 hrs
Score:
0.167
Max. score:
4
Reversibility:
fully reversible within: 72 hrs
Remarks on result:
other: 2/6 animals with score of 1 after 24 hrs
Irritation parameter:
edema score
Basis:
mean
Remarks:
of six animals
Time point:
other: mean of 24 & 72 hrs
Score:
0.167
Max. score:
4
Reversibility:
fully reversible within: 72 hrs
Remarks on result:
other: 2/6 animals with score of 1 after 24 hrs

Erythema and eschar formation

Exposure time (h)

Rabbit 1

Rabbit 2

Rabbit 3

Rabbit 4

Rabbit 5

Rabbit 6

Intact skin

24

0

1

0

0

1

0

do

72

0

0

0

0

0

0

Abraded skin

24

0

1

0

0

1

0

do

72

0

0

0

0

0

0

subtotal

 

0

2

0

0

2

0

Oedma formation

 

 

 

 

 

 

 

Intact skin

24

0

1

0

0

1

0

do

72

0

0

0

0

0

0

Abraded skin

24

0

1

0

0

1

0

do

72

0

0

0

0

0

0

subtotal

 

0

2

0

0

2

0

Total

 

0

4

0

0

4

0

Average

 

0

1.0

0

0

1.0

0

Interpretation of results:
GHS criteria not met
Conclusions:
The substance is not irritating to the skin of rabbbits under the conditions of the study.
Executive summary:

In a study performed to US Ferderal Guidelines the substance tested as a 5% active aqueous dilution was not irritating to the skin of rabbits.

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1997-10-22 to 1997-11-07
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Kulper Rabbitry, Gary, Indiana USA
- Age at study initiation: 8 - 10 weeks
- Weight at study initiation: 2120 - 2370 g
- Housing: stainless steel cages
- Diet (e.g. ad libitum): Purina rabbit Chow ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 4 days

ENVIRONMENTAL CONDITIONS
Environmental conditions were not specified but cages were maintained in a temperature, humidity and light controlled room.

IN-LIFE DATES: From: 1997_10_22 To: 1997_11_6
Type of coverage:
semiocclusive
Preparation of test site:
other: clipped
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL


Duration of treatment / exposure:
4 hours
Observation period:
14 days
Number of animals:
6 male
Details on study design:
TEST SITE
- Area of exposure: 6 cm square
- % coverage: no data
- Type of wrap if used: 2 layer gauze patch held in place with non-irritating Kendall Curity Standard Porous Tape and then covered with a semi-occlusive plastic overwrap secured in place with Kendall Curity Standarf Porous Tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Yes
- Time after start of exposure: 4 hours

SCORING SYSTEM: Draize
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
other: mean of 24, 48 & 72 hrs
Score:
3
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
other: mean of 24, 48 & 72 hrs
Score:
3
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
other: mean of 24, 48 & 72 hrs
Score:
3
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritation parameter:
erythema score
Basis:
animal #4
Time point:
other: mean of 24, 48 & 72 hrs
Score:
3
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritation parameter:
erythema score
Basis:
animal #5
Time point:
other: mean of 24, 48 &72 hrs
Score:
3
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritation parameter:
erythema score
Basis:
animal #6
Time point:
other: mean of 24, 48 & 72 hrs
Score:
3
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritation parameter:
edema score
Basis:
animal #1
Time point:
other: mean of 24, 48 &72 hrs
Score:
2.33
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
edema score
Basis:
animal #2
Time point:
other: mean of 24, 48 & 72 hrs
Score:
1.33
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
edema score
Basis:
animal #3
Time point:
other: mean of 24, 48 & 72 hrs
Score:
1.66
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
edema score
Basis:
animal #4
Time point:
other: mean of 24, 48 & 72 hrs
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
edema score
Basis:
animal #5
Time point:
other: mean of 24, 48 & 72 hrs
Score:
1.66
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
edema score
Basis:
animal #6
Time point:
other: mean of 24, 48 & 72 hrs
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Other effects:
Coriaceousness and slight fissuring on all at 48 hours
Coriaceousness and slight fissuring on all at 72 hours
Some scaling of epidermis on all at 168 hours.

Rabbit

#

Initial bwt (g)

Final bwt (g)

Sex

ER

4.5

ED 4.5

ER

24

ED 24

ER

48

ED 48

ER

72

ED 72

ER

168

ED 168

ER

336

ED 336

403

2040

2260

M

2

1

2

3

3

3

4

1

1

0

0

0

404

2120

2375

M

2

0

2

1

3

2

4

1

2

0

0

0

405

2200

2490

M

1

0

2

2

3

2

4

1

1

0

0

0

406

2070

2250

M

1

1

2

2

3

2

4

2

1

0

0

0

407

2140

2400

M

2

1

2

2

3

2

4

1

2

0

0

0

408

2155

2480

M

1

1

2

1

3

1

4

1

2

0

0

0

 

 

Ave:

1.5

0.67

2.0

1.83

3.0

2.0

4.0

1.17

1.5

0

0

0

 

 

Total

2.17

3.83

5.0

5.17

1.50

0

ER: Erythema

ED: Oedema

Interpretation of results:
Category 2 (irritant) based on GHS criteria
Conclusions:
The substance is highly irritating to the skin of rabbits under the conditions of the study.
Executive summary:

In an OECD 404 Guideline study for acute skin irritation, the substance tested as provided was highly irritating to the skin of rabbits.

Endpoint:
skin irritation: in vitro / ex vivo
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
an in vitro skin irritation study does not need to be conducted because adequate data from an in vivo skin irritation study are available
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Referenceopen allclose all

Endpoint:
eye irritation: in vitro / ex vivo
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
an in vitro eye irritation study does not need to be conducted because adequate data from an in vivo eye irritation study are available
Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
31 August - 22 November, 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
GLP compliance:
yes (incl. certificate)
Species:
cattle
Details on test animals or tissues and environmental conditions:
Bovine eyes from cattle in the age range of 6 to 12 months were obtained from a slaughterhouse . To minimize deterioration and bacterial contamination, on collection the eyes were completely submerged in Hanks’ Balanced Salt Solution (HBSS) containing 1% Penicillin/Streptomycin . Upon arrival at the laboratory the eyes were examined for defects such as but not limited to increased opacity, scratches and neovascularization. Only corneas from eyes free of defects were used.
The corneas were dissected from the eyes so as to leave a 2 to 3 mm rim of sclera and mounted in corneal holders with anterior (epithelium) and posterior (endothelium) chambers. Beginning with the posterior chambers, the chambers were filled to excess with pre-warmed Eagle’s Minimum Essential Medium (EMEM), while preventing bubble formation. The corneal holder was equilibrated at 32±1°C for at least one hour.
After the equilibration period, fresh pre-warmed EMEM was added to both chambers and baseline opacity readings were taken for each cornea. Corneas exhibiting macroscopic tissue damage (e.g. scratches, pigmentation, neovascularisation) or an opacity >7 opacity units were discarded. The mean opacity of all equilibrated corneas was calculated. A minimum of three corneas with opacity values close to the median value for all corneas were selected as negative (or solvent) control corneas. The remaining corneas were then distributed into treatment and positive control groups.
Vehicle:
physiological saline
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750 uL
- Concentration (if solution): 10%

VEHICLE
- Concentration (if solution): 0.9% saline
Duration of treatment / exposure:
10 minutes
Observation period (in vivo):
2 hours
Number of animals or in vitro replicates:
Three corneas were used for each treatment group (test item, negative and positive controls).
Details on study design:
Test groups and treatment:
Three corneas were used for each treatment group (test item, negative and positive controls).

Negative control item: 0.9% NaCl solution
Positive control item: 10% NaOH solution
Test item: 10% solution in 0.9% NaCl solution
Exposure period: 10 minutes

A volume of 750 µL of the test or control items was added to completely cover the cornea’s epithelium in the anterior chamber.
After the exposure period of 10 minutes the exposure solution was removed from each chamber and the epithelium was washed with EMEM containing phenol red (an indicator of pH), at least three times. Washing was repeated until no test item or discolouration (yellow (acid) or purple (alkaline)) of phenol red was visible. The corneas were rinsed a final time with EMEM to remove any remaining phenol red from the chamber. The chamber was then filled with EMEM without phenol red and the corneas were then incubated at 32±1°C for two hours. After this post-exposure incubation period, the corneas were examined.

Examination of the corneas
Corneal injury was assessed by evaluating the opacity and permeability of the cornea. Corneal opacity was determined by the amount of light transmission through the cornea measured quantitatively with the aid of an opacitometer resulting in opacity values measured on a continuous scale.
To determine the corneal permeability 1 mL sodium fluorescein solution (4 mg/mL) was added to the anterior chamber (epithelial surface) and the posterior chamber (endothelial surface) was refilled with fresh EMEM. The holder was incubated in a horizontal position at 32±1°C for 90±5 minutes. The amount of sodium fluorescein that crossed from the anterior to the posterior chamber was measured quantitatively using UV/VIS spectophotometry. Measurements at 492 nm were recorded as optical density (OD492) or absorbance values. The fluorescein permeability values were determined using OD492 values based upon a visible light spectrophotometer using a standard 1 cm path length.

EVALUATION
After correcting the opacity and mean permeability (OD492) values for background opacity and the negative control permeability OD492 values, the mean opacity and permeability OD492 values for each treatment group were combined in an empirically-derived formula to calculate an in vitro irritancy score (IVIS) for each treatment group as follows:
IVIS = mean opacity value + (15 x mean permeability OD492 value)
The opacity and permeability values were also evaluated independently to determine whether the test item induced corrosivity or severe irritation through only one of the two endpoints.

Decision criteria
A test item that induces an IVIS ≥ 55.1 is defined as a corrosive or severe irritant.
As stated in the OECD guideline, if the test substance is not identified as an ocular corrosive or severe irritant, additional testing should be conducted for classification and labeling purposes. The BCOP test method has an overall accuracy of 79% (113/143) to 81% (119/147), a false positive rate of 19% (20/103) to 21% (22/103), and a false negative rate of 16% (7/43) to 25% (10/40), when compared to in vivo rabbit eye test method data classified according to the EPA, EU, or GHS classification systems. When substances within certain chemical (i.e., alcohols, ketones) or physical (i.e., solids) classes are excluded from the database, the accuracy of BCOP across the EU, EPA, and GHS classification systems ranges from 87% (72/83) to 92% (78/85), the false positive rates range from 12% (7/58) to 16% (9/56), and the false negative rates range from 0% (0/27) to 12% (3/26).
Irritation parameter:
in vitro irritation score
Run / experiment:
mean value
Value:
35.1
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: Range = 31.2-40.1

Table 1: Opacity values

 

 

Opacity [Opacity Value]

Mean NC

Corrected Opacity

Mean of group

SD

NaCl 0.9 %

0.7

0.8

-

-

-

0.6

-

1

-

NaOH 10 %

232.9

 

232.1

260.3

41.9

309.2

 

308.4

241.2

 

240.4

Test item 1:10#

25.3

 

24.5

26.6

4.3

24.6

 

23.8

32.3

 

31.5

 

NC: negative control

SD: standard deviation

# tested as 10% concentration in 0.9% NaCl   solution

 

Table 2: Permeability OD Values (492 nm)

 

 

 

Well 1

Well 2

Mean of Duplicates

Mean NC

Corrected OD

Mean of Group

SD

NaCl 0.9 %

0.049

0.052

0.051

0.045

-

-

-

0.038

0.038

0.038

-

0.046

0.046

0.046

-

NaOH 10 %

0.522

0.525

0.524

 

0.479

0.477

0.0

0.507

0.509

0.508

 

0.463

0.54

0.528

0.534

 

0.489

Test item 1:10#

0.441

0.439

0.440

 

0.395

0.511

0.1

0.669

0.661

0.665

 

0.620

0.563

0.563

0.563

 

0.518

 

NC: negative control

SD: standard deviation

# tested as 10% concentration in 0.9% NaCl   solution

 

Table 3: In vitro irritancy score (IVIS)

 

 

NaOH 10 %

Test item 1:10#

Holder 1

240.1

31.2

Holder 2

316.1

33.9

Holder 3

248.5

40.1

Mean

268.3

35.1

SD

41.7

4.5

 

 # tested as 10% concentration in 0.9% NaCl solution

Interpretation of results:
GHS criteria not met
Conclusions:
Based on the results of this study, C10AO is not corrosive to eyes or a severe eye irritant.
Executive summary:

In this study, performed according to OECD 437 under GLP, C10 AO (10% solution in 0.9% NaCl solution) was applied to the epithelial surface of three bovine corneas by addition to the anterior chamber of the corneal holder for an exposure time of ten minutes. Two further groups of three corneas were similarly treated with 0.9% NaCl solution (negative control) or 10% NaOH solution (positive control). Corneal opacity was measured quantitatively as the amount of light transmission through the cornea at 492 nm. Permeability was measured quantitatively as the amount of sodium fluorescein dye that passed across the full thickness of the cornea, as detected in the medium in the posterior chamber. The measurements were used to calculate an in vitro irritancy score (IVIS) which was used to assign an in vitro hazard classification category prediction of the in vivo occular irritation potential of the test item. An opacity value of 26.6 and a permeability value of 0.511 compared to the negative control were determined. An IVIS of 35.1 was calculated. Hence C10 AO (solution) was considered to be non-cytotoxic and not irritant according to the IVIS value, which was <55.1. The test item is not classified as a severe irritant and not corrosive, based on the results of this test.The corneas treated with the positive control (10% NaOH solution) revealed an opacity value of 260.3 and a permeability value of 0.477 compared to the negative control. The IVIS value of 268.3 was well above the cut-off value of 55.1 and hence the acceptance criteria for the test were fulfilled.

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
5 November - 28 November 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
other: ICCVAM-Recommended Test Method Protocol: Hen's Egg Test - Chorioallantoic Membrane (HET-CAM) Test Method (2010)
GLP compliance:
yes (incl. certificate)
Species:
chicken
Strain:
other: White Leghorn
Details on test animals or tissues and environmental conditions:
Fertile white Leghorn chicken eggs
Weight: 48 – 52 g
Age: ≤7 days
Vehicle:
unchanged (no vehicle)
Controls:
yes
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.3 mL/egg
- Concentration (if solution): 40.5% (aqueous solution used without further dilution)

Duration of treatment / exposure:
20 seconds
Observation period (in vivo):
5 minutes
Number of animals or in vitro replicates:
3 eggs/group
Details on study design:
Test item: 3 eggs treated with the test item
Negative control item: 3 eggs treated with physiological saline
Positive control items:3 eggs treated with NaOH (0.1 N); 3 eggs treated with SDS (1%)

Application: on the chorioallantoic membrane of fertilized hens' eggs

SCORING SYSTEM: see tables below
Irritation parameter:
in vitro irritation score
Run / experiment:
Mean of 3 tests
Value:
16.3
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: range 15-17

Table 1: HET-CAM Mean Irritation Scores (IS)

 

Effect according to scoring scheme for irritation testing

Haemorrhage

Lysis

Coagulation

Irritation Score (IS)

C10 AO

 

 

 

Mean

SD

7

3

7

17

7

3

7

17

7

3

5

15

7.0

0.0

3.0

0.0

6.3

1.2

16.3

1.2

Negative control (0.9% NaCl soln)

 

Mean

SD

0

0

0

0

0

0

0

0

0

0

0

0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

Positive control (0.1 N NaOH)

 

Mean

SD

7

3

9

19

7

3

7

17

7

3

7

17

7.0

0.0

3.0

0.0

7.7

1.2

17.7

1.2

Positive control (1% SDS)

 

Mean

SD

7

3

0

10

7

3

0

10

7

3

0

10

7.0

0.0

3.0

0.0

7.0

0.0

10.0

0.0

 

SD: standard deviation

1% SDS: 1% aqueous sodium dodecyl sulfate

Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Conclusions:
Based on the results of this test, C10 AO is considered to be strongly irritating to eyes
Executive summary:

Three eggs each were treated with 0.3 mL C10 AO solution/egg and with the control items. 0.9% NaCl solution was used as the negative control item. 0.1 N Sodium hydroxide (NaOH) or 1% aqueous sodium dodecyl sulphate (SDS) were used as the positive control items. The administration volume for the control items was 0.3 mL per egg. After administration of the test item blood vessels including the capillary system and the albumen were examined and scored for irritant effects (haemorrhage, coagulation and lysis) during 5 minutes. The eggs treated with C10 AO solution revealed an effect with an irritation score (IS) of 16.3. The test item was considered to be a strong irritant.

The positive control items 0.1 N NaOH or 1% SDS caused the expected effect with irritation scores (IS) of 17.7 or 10.0, respectively and, hence, were well within the historical data-range.No effects were observed in the negative control 0.9% NaCl solution.Hence,the HET-CAM assay is considered to be valid.

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
No data
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Principles of method if other than guideline:
Six normal healthy albino rabbits were used to determine whether the test material produces irritation when instilled into rabbits eyes. Each animal had 0.1 mL of the test sample instilled into the right eye with no further treatment. The untreated left eye served as a control Both the treated and control eyes were examined every 24 h for three days.
GLP compliance:
no
Species:
rabbit
Strain:
other: albino
Vehicle:
not specified
Controls:
other: The left eye served as a control.
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit):0.1 mL
- Concentration (if solution): 5 % w/v AO

Observation period (in vivo):
72 hours
Number of animals or in vitro replicates:
Six
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
24/48/72 h
Score:
35.3
Max. score:
110
Reversibility:
not fully reversible within: the observation period
Other effects:
No data
Interpretation of results:
irritating
Conclusions:
Under the conditions of the test, the test substance was a Category 2 eye irritant
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
10-1976
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
according to
Guideline:
other: US Federal Register Vol. 38 (187) 1500:42 (1973).
Deviations:
not specified
GLP compliance:
no
Remarks:
Study pre-dates GLP.
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: No data
- Age at study initiation: 12-14 weeks
- Weight at study initiation: 2.08 kg average
- Housing: caged singly
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17+/-2
- Humidity (%): 50-70
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): natural lighting conditions

Vehicle:
unchanged (no vehicle)
Controls:
other: The untreated eye served as control.
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied: 0.1 mL
- Concentration (if solution): 6 % AO

Duration of treatment / exposure:
Group 1: Eyes remained unwashed for the 72 h test period
Group 2: 5 sec
Observation period (in vivo):
7 days
Number of animals or in vitro replicates:
Group 1: Six
Group 2: Three
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): Group 1 - no washing; Group 2 - 20 mL sterile distilled water at 37°C
- Time after start of exposure: Group 2 - 5 seconds

SCORING SYSTEM: Draize

Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: max score for single animal = 1
Irritation parameter:
iris score
Basis:
mean
Time point:
24/48/72 h
Score:
0.6
Max. score:
4
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: max score for single animal = 1
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
24/48/72 h
Score:
2.5
Max. score:
4
Reversibility:
not fully reversible within: 72 hours
Remarks on result:
other: 4 animals score >=2.7
Irritation parameter:
chemosis score
Basis:
mean
Time point:
24/48/72 h
Score:
2.2
Max. score:
4
Reversibility:
not fully reversible within:
Remarks on result:
other: 3 animals score >=2.3
Other effects:
No data
Interpretation of results:
irritating
Conclusions:
Based on the results of this study the 6 % solution of amine oxide was irritating to the eyes of rabbits.
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1978-05-17
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: As per OECD guidelines 405. Pre-GLP
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
no
Remarks:
Pre-GLP
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Kuiper's Rabbit Ranch, Gary, Indiana
- Age at study initiation: Not available
- Weight at study initiation: 2002 To 2940 gms
- Housing:Individually in hanging wire mesh cages
- Diet (e.g. ad libitum): Purinaw rabbit Chow, ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 6 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): In accordance with recommendations contained in H.E.W. Publication No.74-23 (N.I.H.), entitled"Guide for the care and Use of Laboratory Animals"
- Humidity (%): In accordance with recommendations contained in H.E.W. Publication No.74-23 (N.I.H.),
- Air changes (per hr): In accordance with recommendations contained in H.E.W. Publication No.74-23 (N.I.H.),
- Photoperiod (hrs dark / hrs light):In accordance with recommendations contained in H.E.W. Publication No.74-23 (N.I.H.),


IN-LIFE DATES: From: To:
Vehicle:
water
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml P7270
- Concentration (if solution): Undiluted P7270 applied to the Group I and II, while a 10% (w/w) aqueous solution P7270 was applied to Group III


VEHICLE
- Amount(s) applied (volume or weight with unit): distilled water (for Group III)
- Concentration (if solution): Not applicable
- Lot/batch no. (if required): Not applicable
- Purity: Not applicable
Duration of treatment / exposure:
Group I: 0.1 ml of undiluted P7270 was applied and eyes were not rinsed.
Group II: 0.1 ml of undiluted P7270 was applied and eyes were rinsed with 20 ml lukewarm water approximately for 4 seconds following dosing.
Group III: 0.1 ml of a 10% (w/w) solution of P7270 in distilled water was applied and eyes of animals were not rinsed.
Observation period (in vivo):
After application of test material: 1 hour, 1,2,3,4,7,14,21,28 and 35 days.
Number of animals or in vitro replicates:
3 rabbits of either sex per group.
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): eyes of animals in Group II were rinsed with 20 ml lukewarm water
- Time after start of exposure: approximately for 4 seconds following dosing


SCORING SYSTEM: Draize scoring system


TOOL USED TO ASSESS SCORE: fluorescein
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 1 hour
Score:
6.67
Max. score:
6.67
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Group I: 0.1 ml undiluted test substance, no rinse
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
One animal with grade 1.5 and two with grade 0
Time point:
other: 1 hour
Score:
> 1 - < 1.5
Max. score:
1.5
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Group I: 0.1 ml undiluted test substance, no rinse
Irritation parameter:
iris score
Basis:
mean
Remarks:
all animals with grade 1
Time point:
other: 1 hour, 1 day, 2 days
Score:
1
Max. score:
1
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Group I: 0.1 ml undiluted test substance, no rinse
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
One animal with grade 2, one with grade 1.5 and one with grade 1
Time point:
other: 1 hour
Score:
> 1 - < 2
Max. score:
2
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Group I: 0.1 ml undiluted test substance, no rinse
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
all animals with grade 3
Time point:
other: 1 hour
Score:
3
Max. score:
3
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Group I: 0.1 ml undiluted test substance, no rinse
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 1 day
Score:
7.67
Max. score:
7.67
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Group I: 0.1 ml undiluted test substance, no rinse
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
two animals with grade 2 and one with grade 1
Time point:
other: 1 day
Score:
> 1 - < 2
Max. score:
2
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Group I: 0.1 ml undiluted test substance, no rinse
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
two animals with grade 2 and one with 1.5
Time point:
other: 1 day
Score:
> 1.5 - < 2
Max. score:
2
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Group I: 0.1 ml undiluted test substance, no rinse
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
two animals with grade 3 and one with grade 3.5
Time point:
other: 1 day
Score:
> 3 - < 3.5
Max. score:
3.5
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Group I: 0.1 ml undiluted test substance, no rinse
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 2 days
Score:
7.17
Max. score:
7.17
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Group I: 0.1 ml undiluted test substance, no rinse
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
one animal with grade 2 and two with grade 1
Time point:
other: 2 days
Score:
> 1 - < 2
Max. score:
2
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Group I: 0.1 ml undiluted test substance, no rinse
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
one animal with grade 2 and two with grade 1.5
Time point:
other: 2 days
Score:
> 1.5 - < 2
Max. score:
2
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Group I: 0.1 ml undiluted test substance, no rinse
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
two animals with grade 3 and one with grade 3.5
Time point:
other: 2 days
Score:
> 3 - < 3.5
Max. score:
3.5
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Group I: 0.1 ml undiluted test substance, no rinse
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 3 days
Score:
7.83
Max. score:
7.83
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Group I: 0.1 ml undiluted test substance, no rinse
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
one animal with grade 2, one with 1.5 and one with grade 1
Time point:
other: 3 days
Score:
> 1 - < 2
Max. score:
2
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Group I: 0.1 ml undiluted test substance, no rinse
Irritation parameter:
iris score
Basis:
mean
Remarks:
two animals with grade 1 and one with grade 2
Time point:
other: 3 and 4 days
Score:
> 1 - < 2
Max. score:
2
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Group I: 0.1 ml undiluted test substance, no rinse
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
one animal with grade 2.5, one with 2 and one with grade 1.5
Time point:
other: 3 days
Score:
> 1.5 - < 2.5
Max. score:
2.5
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Group I: 0.1 ml undiluted test substance, no rinse
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
one animal with grade 2.5, one with 3 and one with grade 3.5
Time point:
other: 3 days
Score:
> 2.5 - < 3.5
Max. score:
3.5
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Group I: 0.1 ml undiluted test substance, no rinse
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 4 days
Score:
8
Max. score:
8
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Group I: 0.1 ml undiluted test substance, no rinse
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
one animal with grade 3, one with 1.5 and one animal with grade 1
Time point:
other: 4 days
Score:
> 1 - < 3
Max. score:
3
Reversibility:
not fully reversible within: 35 days
Remarks on result:
other: Group I: 0.1 ml undiluted test substance, no rinse
Irritant / corrosive response data:
Group I: Irreversible irritation upto day 35.
Group II: Irriversible irritation upto day 35.
roup III: All signs of irritation were reversed within 21 days.
Other effects:
Death of one animal of Group III. And the cause of death of animal was unable to determine.
Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Conclusions:
Undiluted P7270 (28% dodecyl dimethyl amine oxide) causes irreversible irritation to rabbit eyes.
Executive summary:

The eyes of three groups of 3 rabbits were instilled with 0.1 ml of P7270 (28% dodecyl dimethyl amine oxide). Group I and II received undiluted P7270 while group III animals were treated with a 10% aqueous solution of P7270 in distilled water. The eyes of the Group I and Group III rabbits were not rinsed and the eyes of the Goup II rabbits were rinsed, approximately 4 seconds following instillation of the test material, with 20 ml of lukewarm distilled water.

Group I: The maximum average score was 8.0 on 4th day. The observable response included irreversible corneal opacity in all three animals till 35 days, iridic response with grade 1 was observed, conjunctival redness upto grade 2 was observed and conjunctival chemosis upto grade 3(swelling with lids about half closed) was observed. The reactions were not fully reversed till 35 days.

Group II: The maximum average score was 6.0 on 3rd day. The observable response included irreversible corneal opacity in one animal till 35 days, iridic response upto grade 1 was observed,conjunctival redness upto grade 2.5 (more diffuse beefy red areas) was observed and conjunctival chemosis upto grade 2.5 was observed which was no fully reversed within 35 days.

Group III: The maximum average score was 4.8 on 2nd day. The observable response included corneal opacity which was reversed within 7 days, iridic response upto grade 1 was observed,conjunctival redness upto grade 2 (more diffuse crimson red areas) was observed and conjunctival chemosis upto grade 2 was observed. All reactions were fully reversed within 21 days in all animals. In this group one rabbit was found dead and the cause of death was unable to determine.

From these observations it can be concluded that P7270 caused irreversible irritation to the rabbit eye. Thus P7270 (28% DDAO) can be classified as GHS category 1.

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Principles of method if other than guideline:
Samples were administered into one eye each of three albino rabbits per test article. The eyes were observed and scored at 1, 24, 48, 72 and 168 hours.
GLP compliance:
no
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Kuiper Rabbitry, Gary, Indiana, USA
- Age at study initiation: 8-10 weeks old
- Weight at study initiation: no data
- Housing: individually housed in stainless steel cages
- Diet: Purina rabbit chow ad libitum
- Water: ad libitum
- Acclimation period: At least 4 days

ENVIRONMENTAL CONDITIONS
The rabbits were maintained according to the recommendations contained in the DHEW Publication No. 86.23 (NIH): 'Guide for the care and Use of Laboratory Animals'.
Vehicle:
unchanged (no vehicle)
Controls:
other: The conlateral eye served as the untreated control for each rabbit
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
- Concentration (if solution): 5% AO (Agent 2108-87-5) 2.5% AO (Agent 2108-87-6)
Duration of treatment / exposure:
Eyes of animals were not rinsed, and observations are noted directly.
Observation period (in vivo):
1, 24, 48, 72 and 168 hours
Number of animals or in vitro replicates:
3
Details on study design:
REMOVAL OF TEST SUBSTANCE
Eyes of animals were not rinsed

SCORING SYSTEM: Draize, JH (1965). Appraisal of the saftey of chemicals in foods, drugs and cosmetics

TOOL USED TO ASSESS SCORE: 2% fluorescein AND uv light
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 24, 48 & 72 hours
Score:
19.4
Max. score:
80
Remarks on result:
other: 5 % AO
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 7 days
Score:
21.6
Max. score:
80
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: 5 % AO
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 24, 48 & 72 hours
Score:
5
Max. score:
10
Remarks on result:
other: 5 % AO
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 7 days
Score:
3.3
Max. score:
10
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: 5 % AO
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
other: 24, 48 & 72 hours
Score:
13.3
Max. score:
20
Remarks on result:
other: 5 % AO
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
other: 7 days
Score:
11.3
Max. score:
20
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: 5 % AO
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 24, 48 & 72 hours
Score:
16.1
Max. score:
80
Remarks on result:
other: 2.5 % AO
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 7 days
Score:
16.7
Max. score:
80
Reversibility:
not fully reversible within: 7 days
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 24, 48 &72 hours
Score:
5
Max. score:
10
Remarks on result:
other: 2.5 % AO
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 7 days
Score:
3.3
Max. score:
10
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: 2.5 % AO
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
other: 24, 48, 72 hours
Score:
12.8
Max. score:
20
Remarks on result:
other: 2.5 % AO
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
other: 7 days
Score:
6.7
Max. score:
20
Reversibility:
not fully reversible within: 7 days
Remarks on result:
other: 2.5 % AO
Interpretation of results:
irritating
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1997-10-23 to 1997-11-13
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Kulper Rabbitry, Gary, Indiana USA
- Age at study initiation: 8 - 10 weeks
- Weight at study initiation: 2120 - 2370 g
- Housing: stainless steel cages
- Diet (e.g. ad libitum): Purina rabbit Chow ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 4 days

ENVIRONMENTAL CONDITIONS
Environmental conditions were not specified but cages were maintained in a temperature, humidity and light controlled room.

IN-LIFE DATES: From: 1997_10_23 To: 1997_11_13
Vehicle:
unchanged (no vehicle)
Controls:
other: untreated eye acted as control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
Duration of treatment / exposure:
21 days (no rinsing)
Observation period (in vivo):
21 days
Number of animals or in vitro replicates:
6
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): No

SCORING SYSTEM: Draize

TOOL USED TO ASSESS SCORE: fluorescein
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
of six animals
Time point:
24/48/72 h
Score:
1.33
Max. score:
4
Reversibility:
not fully reversible within: 21 days
Remarks on result:
other: highest individual mean score = 1.67 (3/6 animals)
Irritation parameter:
iris score
Basis:
mean
Remarks:
of six animals
Time point:
24/48/72 h
Score:
1
Max. score:
2
Reversibility:
not fully reversible within: 21 days
Remarks on result:
other: highest individual mean score =1 (all animals)
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
of six animals
Time point:
24/48/72 h
Score:
3
Max. score:
3
Reversibility:
not fully reversible within: 21 days
Remarks on result:
other: highest individual mean score =3 (all animals)
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
of six animals
Time point:
24/48/72 h
Score:
3
Max. score:
4
Reversibility:
not fully reversible within: 21 days
Remarks on result:
other: highest individual mean score = 3 (all animals)

Group Mean Eye Irritation Scores (Draize Values)

Treatment: No wash

 

Observation Time – Hours

 

1

24

48

72

168

336

504

Averages

 

 

 

 

 

 

 

Cornea

-

20.0

30.0

30.0

43.3

45.0

38.3

Iris

5.0

5.0

5.0

5.0

5.8

6.7

5.8

Conjunctiva

15.7

18.3

18.0

17.0

14.3

11.7

7.0

Total

20.7

43.3

53.0

52.0

63.4

63.4

51.1

Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Executive summary:

In an OECD 405 Guideline study for acute eye irritation, the substance tested as provided produced irreversible effects on the eye and is classified as Category 1.

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Qualifier:
according to
Guideline:
other: CFR 16:1500.42
GLP compliance:
not specified
Species:
rabbit
Strain:
other: albino
Details on test animals or tissues and environmental conditions:
no data
Controls:
other: untreated left eye used as control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
- Concentration (if solution): 2.5 %w/w

Duration of treatment / exposure:
72 hours
Observation period (in vivo):
72 hours
Number of animals or in vitro replicates:
6
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): Not applicable
- Time after start of exposure: Not applicale

SCORING SYSTEM: Draize
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
(of 6 animals)
Time point:
other: mean of 24, 48 & 72 hrs
Score:
0.11
Max. score:
4
Reversibility:
fully reversible within: 72 hrs
Remarks on result:
other: highest individual mean score = 0.67 (1 animal)
Irritation parameter:
iris score
Basis:
mean
Remarks:
(of 6 animals)
Time point:
other: mean of 24, 48 & 72 hrs
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
(of 6 animals)
Time point:
other: mean of 24, 48 & 72 hrs
Score:
0.89
Max. score:
3
Reversibility:
not fully reversible within: 72 hrs
Remarks on result:
other: highest individual mean score = 1.0 (4/6 animals)
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
(of 6 animals)
Time point:
other: mean of 24, 48 & 72 hrs
Score:
0.44
Max. score:
4
Reversibility:
not fully reversible within: 72 hrs
Remarks on result:
other: highest individual mean score = 1.67 (1/6 animals)

 

Observation Time – Hours

 

24

48

72

Averages

 

 

 

Cornea

0.16

0.16

0

Iris

0

0

0

Conjunctiva

1

1

0.67

Chemosis

0.67

0.5

0.16

Interpretation of results:
GHS criteria not met
Conclusions:
A 2.5 % w/w solution of C14 AO was not irritating to the eyes of rabbits in a study performed according to CFR 16:1500.42.
Executive summary:

In a study performed according to CFR 16:1500.42, a 2.5 % w/w solution of C14 AO in water was not irritating to the eyes of rabbits.

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Meets generally accepted scientific standards with acceptable restrictions.
Qualifier:
according to
Guideline:
other: CFR 16 1500.42
GLP compliance:
not specified
Species:
rabbit
Vehicle:
water
Controls:
other: untreated left eye used as control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
- Concentration (if solution): diluted to 2.5% active

Duration of treatment / exposure:
7 days
Observation period (in vivo):
7 days
Number of animals or in vitro replicates:
6
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): not applicable
- Time after start of exposure: not applicable

SCORING SYSTEM: Draize
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
(of 6 animals)
Time point:
other: mean of 24, 48 & 72 hrs
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
mean
Remarks:
of 6 animals
Time point:
other: mean of 24, 48 & 72 hrs
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
of 6 animals
Time point:
other: mean of 24, 48 & 72 hrs
Score:
0.167
Max. score:
3
Reversibility:
fully reversible within: 48 hrs
Remarks on result:
other: highest individual mean score = 0.33 (3/6 animals)
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
(of 6 animals)
Time point:
other: mean of 24, 48 & 72 hrs
Score:
0.167
Max. score:
4
Reversibility:
fully reversible within: 48 hrs
Remarks on result:
other: highest individual mean score = 0.33 (3/6 animals)

Group test summary

Group 1 (3 animals)

 

Observation Time – Hours

 

24

48

72

96

7-days

Cornea

0

0

0

0

0

Iris

0

0

0

0

0

Conjunctiva

6

0

0

0

0

Average Daily score

2.0

0

0

0

0

Group 2 (3 animals)

 

Observation Time – Hours

 

24

48

72

96

7-days

Cornea

0

0

0

0

0

Iris

0

0

0

0

0

Conjunctiva

12

0

0

0

0

Average Daily score

4.0

0

0

0

0

Interpretation of results:
GHS criteria not met
Conclusions:
The substance (a 2.5 % solution of C14 AO) is not irritating to the eye of rabbbits under the conditions of the study.
Executive summary:

In a study performed to US Federal Guidelines the substance tested as a 2.5% active aqueous dilution was not irritating to the eyes of rabbits.

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1978-09-12 To 1978-10-18
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
no
Remarks:
Pre-GLP
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Kings Wheel
- Age at study initiation: Young adult
- Weight at study initiation: Males: 1608.6 gm, Females: 2031.5 gm
- Housing:Individually in stainless steel cages with stainless steel slatted flooring
- Diet (e.g. ad libitum): Purins Lab Rabbit Chow (By Ralston Purina Company)
- Water (e.g. ad libitum): Tap water, from bottles with tubules made of refined steel, ad libitum
- Acclimation period: minimum 7 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-22
- Humidity (%): 46-72
- Air changes (per hr): Not available
- Photoperiod (hrs dark / hrs light): 12/12 (Fluorescent lighting)


IN-LIFE DATES: From: 1978-09-12 To: 1978-10-18
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml P0434
- Concentration (if solution): Undiluted


VEHICLE
- Amount(s) applied (volume or weight with unit): Not applicable
- Concentration (if solution): Not applicable
- Lot/batch no. (if required): Not applicable
- Purity: Not applicable
Duration of treatment / exposure:
Group I: Eyes of animals were not rinsed, and observations are noted directly.
Group II: Eyes of animals were rinsed after 4 seconds by spraying 20 ml of lukewarm tap water after application.
Observation period (in vivo):
At 1 hour, 1,2,3,4,7,14,21,28 and 35 days.
Number of animals or in vitro replicates:
3 animals of either sex per group.
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): Eyes of Group II animals were rinsed by spraying 20 ml of lukewarm tap water from a hypodermic syringe fitted with a snubbed 18 gauge needle into the eye under moderate pressure.
- Time after start of exposure: 4 seconds after application.


SCORING SYSTEM: Draize scoring system


TOOL USED TO ASSESS SCORE: A Pen light
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
animal #1
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
not reversible
Remarks:
till day 35. Vascularization was observed from day 21 onwards till the end of study (day 35)
Remarks on result:
other: No rinse
Irritation parameter:
iris score
Basis:
mean
Remarks:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: Iris was normal throughout the study.
Remarks on result:
other: No rinse
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
animal #1
Time point:
24/48/72 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: No rinse
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
animal #1
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: No rinse
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
animal #2
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
not reversible
Remarks:
till day 35. Vascularization was observed from day 21 onwards till the end of study (day 35)
Remarks on result:
other: No rinse
Irritation parameter:
iris score
Basis:
mean
Remarks:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: Iris was normal throughout the study.
Remarks on result:
other: No rinse
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
animal #2
Time point:
24/48/72 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: No rinse
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
animal #2
Time point:
24/48/72 h
Score:
1.33
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: No rinse
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
animal #3
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
not reversible
Remarks:
till day 35. Vascularization was observed from day 21 onwards till the end of study (day 35)
Remarks on result:
other: No rinse
Irritation parameter:
iris score
Basis:
mean
Remarks:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: Iris was normal throughout the study.
Remarks on result:
other: No rinse
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
animal #3
Time point:
24/48/72 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: No rinse
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
animal #3
Time point:
24/48/72 h
Score:
1.33
Max. score:
3
Reversibility:
fully reversible within: 14 days
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
animal #4
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: Cornea was normal throughout the study
Remarks on result:
other: Rinse
Irritation parameter:
iris score
Basis:
mean
Remarks:
animal #4
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: Iris was normal throughout the study.
Remarks on result:
other: Rinse
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
animal #4
Time point:
24/48/72 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: Rinse
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
animal #5
Time point:
24/48/72 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 14 days
Remarks on result:
other: Rinse
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
animal #4
Time point:
24/48/72 h
Score:
0.67
Max. score:
4
Reversibility:
fully reversible within: 3 days
Remarks on result:
other: Rinse
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
animal #5
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: Rinse
Irritation parameter:
iris score
Basis:
mean
Remarks:
animal #5
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: Iris was normal throughout the study.
Remarks on result:
other: Rinse
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
animal #5
Time point:
24/48/72 h
Score:
0.67
Max. score:
4
Reversibility:
fully reversible within: 3 days
Remarks on result:
other: Rinse
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
animal #6
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
not reversible
Remarks:
till day 35. Vascularization was observed from day 21 onwards till the end of study (day 35)
Remarks on result:
other: Rinse
Irritation parameter:
iris score
Basis:
mean
Remarks:
animal #6
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: Iris was normal throughout the study.
Remarks on result:
other: Rinse
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
animal #6
Time point:
24/48/72 h
Score:
1
Max. score:
20
Reversibility:
fully reversible within: 14 days
Remarks:
Vascularization was observed from day 21 onwards till the end of study (day 35)
Remarks on result:
other: Rinse
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
animal #6
Time point:
24/48/72 h
Score:
0.67
Max. score:
4
Reversibility:
fully reversible within: 3 days
Remarks on result:
other: Rinse
Other effects:
None

Table 1: The eye irritation response of animals (Group I - No rinse) exposed to undiluted P0434 (27.8% DDAO) (study# 21599)

Animal number Observation time Cornea Cornea Score Iris Iris Score Conjunctivae Conjunctivae Score
A (Degree of density) B (Area of cornea involved) AxBx5 A Ax5 A (Redness) B (Chemosis) C (Discharge) A+B+Cx2
Animal #1 1 hour 0 0 0 0 0 1 2 3 12
Day 1 1 4 20 0 0 1 2 3 12
Day 2 1 4 20 0 0 1 2 2 10
Day 3 1 4 20 0 0 1 2 2 10
Day 4 1 4 20 0 0 1 2 1 8
Day 7 1 4 20 0 0 1 1 1 6
Day 14 1 4 20 0 0 0 0 0 0
Day 21* 1 4 20 0 0 0 0 0 0
Day 28* 2 4 40 0 0 0 0 0 0
Day 35* 2 4 40 0 0 0 0 0 0
                     
Animal #2 1 hour 0 0 0 0 0 1 2 3 12
Day 1 1 4 20 0 0 1 2 2 12
Day 2 1 4 20 0 0 1 1 1 6
Day 3 1 4 20 0 0 1 1 1 6
Day 4 1 4 20 0 0 1 0 0 2
Day 7 1 4 20 0 0 1 0 0 2
Day 14 1 4 20 0 0 0 0 0 0
Day 21* 1 4 20 0 0 0 0 0 0
Day 28* 2 2 20 0 0 0 0 0 0
Day 35* 2 2 20 0 0 0 0 0 0
                     
Animal #3 1 hour 0 0 0 0 0 1 1 2 8
Day 1 1 4 20 0 0 1 2 1 8
Day 2 1 4 20 0 0 1 1 1 6
Day 3 1 4 20 0 0 1 1 1 6
Day 4 1 4 20 0 0 1 1 1 6
Day 7 1 4 20 0 0 1 1 1 6
Day 14 1 4 20 0 0 0 0 0 0
Day 21* 1 4 20 0 0 0 0 0 0
Day 28* 1 4 20 0 0 0 0 0 0
Day 35* 1 4 20 0 0 0 0 0 0

* Vascularization was observed. Vascularization refers to the invasion of the cornea by blood vessels.

Table 2: The eye irritation response of animals (Group II- Rinse) exposed toundiluted P0434 (27.8% DDAO) (study# 21599)

Animal number Observation time Cornea Cornea Score Iris Iris Score Conjunctivae Conjunctivae Score
A (Degree of density) B (Area of cornea involved) AxBx5 A Ax5 A (Redness) B (Chemosis) C (Discharge) A+B+Cx2
Animal #4 1 hour 0 0 0 0 0 1 2 3 12
Day 1 0 0 0 0 0 1 2 1 8
Day 2 0 0 0 0 0 1 0 1 4
Day 3 0 0 0 0 0 1 0 1 4
Day 4 0 0 0 0 0 1 0 0 2
Day 7 0 0 0 0 0 1 0 0 2
Day 14 0 0 0 0 0 0 0 0 0
                     
Animal #5 1 hour 0 0 0 0 0 1 2 2 10
Day 1 1 1 5 0 0 1 2 1 8
Day 2 1 1 5 0 0 1 0 1 4
Day 3 1 1 5 0 0 1 0 1 4
Day 4 1 1 5 0 0 1 0 0 2
Day 7 0 0 0 0 0 1 0 0 2
Day 14 0 0 0 0 0 0 0 0 0
                     
Animal #6 1 hour 0 0 0 0 0 1 2 3 12
Day 1 1 2 10 0 0 1 2 1 8
Day 2 1 2 10 0 0 1 0 1 4
Day 3 1 2 10 0 0 1 0 2 6
Day 4 1 2 10 0 0 1 0 2 6
Day 7 1 3 15 0 0 1 0 2 6
Day 14 1 2 10 0 0 0 0 0 0
Day 21* 1 2 10 0 0 0 0 1 2
Day 28* 1 2 10 0 0 0 0 1 2
Day 35* 1 2 10 0 0 0 0 0 0

* Vascularization was observed. Vascularization refers to the invasion of the cornea by blood vessels.

Overall irritation/ Corrosion results: For Rinse Group

Irritation parameter  Basis  Time point    Max.score  Reversibility  Remarks 
Overall irritation score  mean 1 hour   3.0 3.0  not fully reversible within:35 days  Rinse 
Cornea score  mean: all animals with grade 0  1 hour  not fully reversible within:35 days  Rinse 
Iris score    1 hour, 1,2,3,4,7,14,21,28 and 35 days  No iridal response was observed.  Rinse 
Conjunctival redness score  mean: all animals with grade 1 1 hour, 1,2,3,4 and 7 days  fully reversible within:14 days  Rinse 
Conjunctival chemosis score  mean  1 hour and day 1  fully reversible within: day 2  Rinse 
Overall irritation score  mean  day 1  3.67  3.67  not fully reversible within:35 days  Rinse 
Cornea score  mean: Two animals with grade 1 and one with grade 0 day 1,2,3,4  >0 -<1  not fully reversible within:35 days  Rinse 
Overall irritation score  mean  days 2,3,4  1.67  1.67  not fully reversible within:35 days  Rinse 
Conjunctival chemosis score  mean: all animals with grade 0  days 2,3,4,7,14,21,28 and 35 days   0 fully reversible within:2 days  Rinse 
Overall irritation score mean  day 7  1.33  1.33  not fully reversible within:35 days  Rinse 
Cornea score mean:one animal with grade 1 and two eith grade 0  day 7, 14  >0 - <1   not fully reversible within:35 days Rinse 
Conjunctival redness score  mean:all animals with grade 0  day 14,21,28 and 35  0  0  fully reversible within:14 days Rinse 
Overall irritation score  mean  day 14, 21,28 and 35  0.33   0.33  not fully reversible within:35 days Rinse 
Cornea score  mean:one animals with grade 1 and two with grade 0  day 14, 21, 28 and 35  >0 - <1   1 not fully reversible within:35 days  Rinse 

GHS Classification:

Animal #  GHS Classification  What it means? Cornea  Iris  Conjunctival redness Conjunctival chemosis  Days to clear by individual animal 
 No rinse (Group I)              
 1 Irreversible eye irritation  1.0   1.0 2.0  Cornea not reversible by 35 days 
 2 Irreversible eye irritation  1.0  0  1.0 1.33  Cornea not reveersible by 35 days 
Irreversible eye irritation  1.0   1.0 1.33 Cornea not reversible by 35 days 
Rinse (Group II)               
 1  1 Irreversible eye irritation  0.0   0  1.0  0.67 Fully reversible within 14 days 
 2  1 Irreversible eye irritation   1.0  0 1.0   0.67 Fully reveersible within 14 days 
 3  1 Irreversible eye irritation   1.0  0  1.0 0.67  Cornea not fully reversible by 35 days 
Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Remarks:
Migrated information
Conclusions:
P0434 (27.8% Dodecyl dimethyl amine oxide) causes irreversible irritation to the rabbit eye.
Executive summary:

The eyes of two groups of 3 rabbits were instilled with 0.1 ml of undiluted P0434 (27.8% DDAO). The eyes of the first group of 3 rabbits were not rinsed and the eyes of the second group of 3 rabbits were rinsed, approximately 4 seconds following instillation of the test material, with 20 ml of lukewarm distilled water.

Group I: The maximum average score was 4.0 at 1day. The observable response included irreversible corneal opacity in all three animals till 35 days, no iridic response was observed, conjunctival redness with grade 1 was observed which was fully reversed within 14 days and conjunctival chemosis upto grade 2 was observed which was fully reversed within 14 days.

Group II: The maximum average score was 3.67 at one day. The observable response included irreversible corneal opacity in one animal till 35 days, no iridic response was observed,conjunctival redness with grade 1 was observed which was fully reversed within 14 days and conjunctival chemosis up to grade 2 was observed which was fully reversed within 2 days.

From these observations it can be concluded that the test material P0434 (27.8% dodecyl dimethyl amine oxide) is causing irreversible irritation to the rabbit eye. Thus 27.8% DDAO can be classified as GHS category 1.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation:

In the key study, performed according to OECD TG 404 [Fulfs JC (1978)] using C12-14 AO, the substance (0.4 mL, 27.8 %AO) was applied to the intact and abraded skin of 3 female rabbits (New Zealand White) and covered with an occlusive dressing for 24 hours. At the end of the exposure period the treatment sites were wiped with a wet paper towel and reactions noted over a 72 hour period. Very slight to well defined erythema was observed at the intact and abraded sites of all the test animals at 24 hours. At 72 hours severe erythema (beet redness) to slight eschar formation were observed at the intact and abraded sites of all test animals. No oedema was observed at any time point. Similar results were obtained in a supporting study [Dean WP (1978)].

In three further supporting studies available for C12-14 AO, performed according to US Federal Register, vol 41(188): 42572 (27th September 1976), the substance (0.5 mL, 30 % AO) was applied to the intact skin of 6 rabbits (albino) and covered with an occlusive dressing for 4 hours. At the end of the exposure period the test substance was removed by wiping with a paper towel moistened with sterile distilled water. The sites were scored immediately and after 48 hours. Erythema and oedema scores increased during the 48 hour observation period. No destruction of intact skin was observed after the four-hour exposure.

Three further studies are available performed using lower concentrations of C12-14 AO. In the first of these [Nicholas P & Jones JR (1976)] performed according to US Federal Register Vol 38 (187): 1500:41 (1973) the substance (0.5 mL, 6 %w/w AO) was applied to the intact and abraded skin of 6 rabbits (New Zealand White) and covered with an occlusive dressing for 24 hours. Very slight to moderate to severe erythema and very slight to slight oedema was observed at the intact and abraded sites of all test animals at 24 hours. At 72 hours no to slight erythema and no to very slight oedema was observed at the intact and abraded sites of all test animals. In the second study the substance (0.5 mL, 0.6 % AO) was applied to the intact and abraded skin of 8 rabbits (New Zealand White) and covered with an occlusive dressing for 24 hours. No to very slight erythema and oedema was observed at the intact and abraded sites of all test animals at 24 and 72 hours. In the third study [Kukulinski M (1997)] performed according to FHSA/CPSC Design, 16 CFR1500 the substance (0.5 mL, 2.5 or 5 %AO) was applied to the intact and abraded skin of 3 rabbits (New Zealand White). At 5 % AO one rabbit had no erythema at 24 hours on had very slight and the third had well defined erythema at the intact sites. At 72 hours two rabbits had no erythema and the third very slight erythema. No oedema was observed at either 24 or 72 hours. At 2.5 % AO very slight erythema was observed in one rabbit at 24 hours. All other scores were zero at both 24 and 72 hours.

In addition, supporting studies are available for the following category members:

C10 AO: in vitro skin corrosivity (OECD TG 431) – not corrosive [Flugge C (2012)]; in vitro skin irritation (OECD TG 439) – not irritant [Flugge C (2012)]; in vivo skin irritation in rabbits (OECD TG 404) – not irritant [Leuschner J (2012)].

C14 AO: in vivo skin irritation in rabbits (OECD TG 404] – irritant (Category 2) [Kukulinski M (1997)]; in vivo skin corrosivity in rabbits (US FR, vol 41(188): 42572) – not corrosive [Haynes G (1982)] in vivo skin irritation in rabbits (CFR 16.1500.41) – 5% solution not irritating [Lacap & Frances (1986)].

C12-18 AO: in vivo skin irritation in rabbits (OECD TG 404) – irritant (Category 2) [Hollander & Rupprich (1983)].

Eye irritation

In the key study, performed according to OECD TG 405 [Fulfs JC (1978)] using C12-14 AO, the test substance (0.1 mL, 27.8 % AO) was instilled into the eyes of two groups of three rabbits (New Zealand White). The eyes of the first group of 3 rabbits were not rinsed and the eyes of the second group of 3 rabbits were rinsed, approximately 4 seconds following instillation of the test substance, with 20 ml of lukewarm distilled water. In the first group the maximum average score was 4.0 at one day. The observable response included irreversible corneal opacity in all three animals up to 35 days. No iridic response was observed. Conjunctival redness with grade 1 and conjunctival chemosis up to grade 2 were observed and fully reversed within 14 days. In the second group the maximum average score was 3.67 at one day. The observable response included irreversible corneal opacity in one animal up to 35 days. No iridic response was observed. Conjunctival redness with grade 1 and conjunctival chemosis up to grade 2 were observed and fully reversed within 2 days. From these observations it is concluded that the test substance caused irreversible damage to the rabbit eye.

In a supporting study performed according to OECD TG 405 [Dean WP (1978)] using C12-14 AO, the test substance was instilled into the eyes of two groups of three rabbits (New Zealand White). Group I and Group II received 0.1 mL of 28 % AO. Group III received 0.1 mL of a 10 % dilution, i.e. 2.8 % AO. The eyes of the Group I and Group III rabbits were not rinsed and the eyes of the Group II rabbits were rinsed, approximately 4 seconds following instillation of the test material, with 20 mL of lukewarm distilled water. For Group I the maximum average score was 8.0 on 4th day. The observable response included irreversible corneal opacity in all three animals till 35 days. Iridic response with grade 1 was observed. Conjunctival redness up to grade 2 was observed and conjunctival chemosis up to grade 3 (swelling with lids about half closed) was observed. The reactions were not fully reversed up to 35 days. For Group II the maximum average score was 6.0 on the third day. The observable response included irreversible corneal opacity in one animal up to 35 days. Iridic response up to grade 1 was observed. Conjunctival redness up to grade 2.5 (more diffuse beefy red areas) and conjunctival chemosis up to grade 2.5 were observed which were not fully reversed within 35 days. For Group III the maximum average score was 4.8 on the second day. The observable response included corneal opacity which was reversed within 7 days. Iridic response up to grade 1 was observed. Conjunctival redness up to grade 2 (more diffuse crimson red areas) and conjunctival chemosis up to grade 2 were observed. All reactions were fully reversed within 21 days in all animals. In this group one rabbit was found dead and the cause of death was unable to be determined. From these observations it can be concluded that the test substance caused irreversible damage to the rabbit eye. A 10 % dilution of the test substance (i.e. 2.8 % AO) caused irritation that was reversible within 21 days.

Two further studies are available performed on lower concentrations of C12-14 AO. In both of these studies, which used 6 % AO [Nicholas P & Jones JR (1976)] and 5 % or 2.5 % AO [Kukulinski M (1997)] the test substance induced eye irritation effects that were not fully reversible within a 7 -day period. It is not possible to conclude for certain from the 7 -day scores whether the effects would be fully reversed if the observation period had been extended to 21 days, however based on the study performed with 2.8 % AO this seems likely.

In addition, supporting studies are available for the following category members:

C10 AO: in vitro eye irritation (OECD 437) – not irritant [Leuschner J (2012)]; in vitro eye irritation (HET-CAM) – Category 1 [Haferkorn J (2012)].

C14 AO: in vivo eye irritation in rabbits (OECD TG 405) – Category 1 [Kukulinski M (1997)]; in vitro eye irritation in rabbits (CFR 16 1500.42) – 2.5% solution not irritating [Lacap & Frances (1986)]; in vitro eye irritation in rabbits (CFR 16 1500.42) – 2.5% solution not irritating [Rothstein & Platt (1985)].

C12-18 AO: in vivo eye irritation in rabbits (similar to OECD TG 405) – 5% solution Category 2 irritant [Bullens P (1984)].

Justification for classification or non-classification

The substance, C12 -14 AO, is classified as Skin Irritant Category 2 on the basis of the reactions seen in the skin irritation studies.

The substance, C12 -14 AO, caused damage to the eyes of rabbits that did not fully reverse during the 21 day observation period and is therefore classified as Eye Damage Category 1.