Amines, C12-14 (even numbered)-alkyldimethyl, N-oxides

Endpoint summary

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

Reference

Reference 1

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

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)

GLP compliance:

yes (incl. QA statement)

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.

Reference 2

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

Guideline:

other: OECD Guideline No. 439

Qualifier:

according to guideline

Guideline:

other: EC method B46

GLP compliance:

yes (incl. QA statement)

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.

 

Reference 3

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

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

Himalayan

Details on test animals or test system 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.

Reference 4

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

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.

Reference 5

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

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.

Reference 6

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

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 or test system 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.

Reference 7

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

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.

Reference 8

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

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 or test system 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.

Reference 9

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

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 or test system 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.

Reference 10

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

Guideline:

other: FHSA/CPSC Design, 16 CFR 1500

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system 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.

Reference 11

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

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system 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.

Reference 12

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

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.

Reference 13

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

Guideline:

other: US Federal Register, Vol 41, part 188 p42572 1976

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system 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.

Reference 14

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

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.

Reference 15

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

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system 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.

Reference 16

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

Reference

Reference 1

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

Reference 2

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

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

GLP compliance:

yes (incl. QA statement)

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.

Reference 3

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

Guideline:

other: ICCVAM-Recommended Test Method Protocol: Hen's Egg Test - Chorioallantoic Membrane (HET-CAM) Test Method (2010)

GLP compliance:

yes (incl. QA statement)

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.

Reference 4

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

Reference 5

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

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.

Reference 6

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

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.

Reference 7

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

Reference 8

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

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

GLP compliance:

yes (incl. QA statement)

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.

Reference 9

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

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.

Reference 10

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

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.

Reference 11

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

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	0	0	not fully reversible within:35 days	Rinse
Iris score		1 hour, 1,2,3,4,7,14,21,28 and 35 days	0	0	No iridal response was observed.	Rinse
Conjunctival redness score	mean: all animals with grade 1	1 hour, 1,2,3,4 and 7 days	1	1	fully reversible within:14 days	Rinse
Conjunctival chemosis score	mean	1 hour and day 1	2	2	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	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	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	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	1	Irreversible eye irritation	1.0	0	1.0	2.0	Cornea not reversible by 35 days
2	1	Irreversible eye irritation	1.0	0	1.0	1.33	Cornea not reveersible by 35 days
3	1	Irreversible eye irritation	1.0	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.