Registration Dossier

Administrative data

Description of key information

- Skin irritation: OECD 404, 6 (3m/3f) New Zealand White rabbits, very slight to moderate erythema (all animals), very slight oedema (one animal), fully reversible. Conclusion: slightly irritating, no C&L.
- Eye irritation: OECD 405, 3 New Zealand White rabbits, no corneal opacity, iridial inflammation in one animal at 1-hour reading time point and conjunctival irritation in six animals (mean of gradings: 2.0) . Conclusion: irritating to eyes.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation / corrosion, other
Remarks:
in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1999-05-27 to 1999-06-09
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Version / remarks:
the European Economic Communities (EEC) Guideline in the Official Journal of the European Communities [92/69, Annex V, B4 ( 1992)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2500 (Acute Dermal Irritation)
Deviations:
no
GLP compliance:
yes
Remarks:
the United States Environmental Protection Agency (EPA) Good Laboratory Practice Regulations (40 CFR Parts 160 and 792)
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Covance Research Products, Inc., Denver, PA
- Age at study initiation: Young adult
- Weight at study initiation: 2523 to 3833 grams at initiation of dosing
- Housing: Individual in suspended wire-mesh cages
- Diet: PMI Nutrition International, Inc. Certified Rabbit LabDiet@ 5322 was offered at approximately 150 g per day during the study.
- Water (e.g. ad libitum): Reverse osmosis-treated municipal water was provided ad libitum.
- Acclimation period: for a minimum of 7 days prior to initiation of dosing.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 64.1- 67.2 °F (18 - 20 °C)
- Humidity (%): 50.1-76.7 The humidity was slightly above the protocol-specified range (30-70%) four times within the study period. A brief period of slightly increased humidity would not be expected to adversely affect the health of the rabbits. Therefore, this deviation did not affect the quality or integrity of the studv.
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): not reported
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
Prior to use, the original container of the test article was agitated to insure a homogeneous mixture. A sufficient amount of test article was dispensed and a stir bar was added. The test article was stirred continuously throughout use.
- Amount(s) applied (volume or weight with unit): 0.5 mL /site
Duration of treatment / exposure:
4 hours
Observation period:
approximately 30-60 minutes and 24, 48 and 72 hours after patch removal and daily thereafter for up to 13 days
Number of animals:
Three males and three females
Details on study design:
TEST SITE
On the day prior to dosing, the hair was removed from the backs and flanks of the rabbits using an electric clipper. Each 0.5 mL dose was applied to an area of skin approximately 2.5 x 2.5 cm under a two-ply gauze patch secured in place with Micropore@ tape. The patches were overwrapped with a gauze binder and secured with Dermiform@ tape. Plastic restraint collars were applied to the animals to prevent ingestion of the test article andor bandages.
- Area of exposure: an area of skin approximately 2.5 x 2.5 cm on backs and flanks
- Type of wrap if used: Plastic restraint collars

REMOVAL OF TEST SUBSTANCE
At the end of four hours, the collars and bandages were removed and the sites wiped with disposable paper towels moistened with deionized water.

SCORING SYSTEM (in accordance with the method of Draize):
Erthema and Eschar Formation:
- No erythema: 0
- Very slight erythema (barely perceptible, edges of area not well defined): 1
- Slight erythema (pale red color and edges definable): 2
- Moderate to severe erythema (definite red in color and area well defined): 3
- Severe erythema (beet or crimson red) to slight eschar formation (injuries in depth): 4
- Maximum possible erythema score: 4

Oedema Formation:
- No oedema: 0
- Very slight oedema (barely perceptible, edges of area not well defined): 1
- Slight oedema (edges of area well defined by definite raising): 2
- Moderate oedema (raised approximately 1 mm): 3
- Severe oedema (raised more than 1 mm and extending beyond area of exposure): 4
- Maximum possible oedema score: 4

Maximum total possible Primary Irritation Score: 8

DESCRIPTIVE RATINGS
Mean Primary Dermal Irritation Index:
- Nonirritating: 0
- Slightly Irritating: 0.1 - 2.0
- Moderately Irritating: 2.1 - 5.0
- Severely Irritating: 5.1 - 8.0

Draize, J.H., 1965. The Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics. Dermal Toxicity, pp. 46-59. Assoc. Of Food and Drug Officials of the U.S., Topeka, Kansas and the EPA-OPPTS Health Effects Test Guidelines (1998).

Irritation parameter:
primary dermal irritation index (PDII)
Basis:
mean
Time point:
other: 1h, 24h, 48h and 72h
Score:
1.4
Max. score:
8
Reversibility:
fully reversible within: 13 days
Remarks on result:
other: slightly irritating in accordance with the descriptive rating of Draize
Irritation parameter:
erythema score
Basis:
animal #1
Remarks:
male
Time point:
other: mean of 24, 48 and 72-h
Score:
1.67
Max. score:
2
Reversibility:
fully reversible within: 11 days
Remarks on result:
other: brown staining on dose site (24 and 48 h); desquamation on days 5-10
Irritation parameter:
erythema score
Basis:
animal #2
Remarks:
male
Time point:
other: mean of 24, 48 and 72-h
Score:
1
Max. score:
2
Reversibility:
fully reversible within: 8 days
Remarks on result:
other: brown staining on dose site (24 and 48 h); desquamation on days 6-7
Irritation parameter:
erythema score
Basis:
animal #3
Remarks:
male
Time point:
other: mean of 24, 48 and 72-h
Score:
0.67
Max. score:
1
Reversibility:
fully reversible within: 4 days
Remarks on result:
other: brown staining on dose site (24 h)
Irritation parameter:
erythema score
Basis:
animal #4
Remarks:
female
Time point:
other: mean of 24, 48 and 72-h
Score:
2.33
Max. score:
3
Reversibility:
fully reversible within: 13 days
Remarks on result:
other: brown staining on dose site (24 h); desquamation on days 4-11
Irritation parameter:
erythema score
Basis:
animal #5
Remarks:
female
Time point:
other: mean of 24, 48 and 72-h
Score:
2
Max. score:
3
Reversibility:
fully reversible within: 11 days
Remarks on result:
other: brown staining on dose site (24 and 48 h); desquamation on days 4-10
Irritation parameter:
erythema score
Basis:
animal #6
Remarks:
female
Time point:
other: mean of 24, 48 and 72-h
Score:
1.67
Max. score:
2
Reversibility:
fully reversible within: 8 days
Remarks on result:
other: brown staining on dose site (24 and 48 h); desquamation on days 6-7
Irritation parameter:
edema score
Basis:
animal #1
Remarks:
male
Time point:
other: mean of 24, 48 and 72 h
Score:
0
Max. score:
0
Irritation parameter:
edema score
Basis:
animal #2
Remarks:
male
Time point:
other: mean of 24, 48 and 72 h
Score:
0
Max. score:
0
Irritation parameter:
edema score
Basis:
animal #3
Remarks:
male
Time point:
other: mean of 24, 48 and 72 h
Score:
0
Max. score:
0
Irritation parameter:
edema score
Basis:
animal #4
Remarks:
female
Time point:
other: mean of 24, 48 and 72 h
Score:
0.67
Max. score:
1
Reversibility:
fully reversible within: 6 days
Irritation parameter:
edema score
Basis:
animal #5
Remarks:
female
Time point:
other: mean of 24, 48 and 72 h
Score:
0
Max. score:
0
Irritation parameter:
edema score
Basis:
animal #6
Remarks:
female
Time point:
other: mean of 24, 48 and 72 h
Score:
0
Max. score:
0
Irritant / corrosive response data:
The test article induced very slight to moderate erythema on all animals and very slight oedema for one rabbit. Desquamation and brown staining of the skin were noted for five and six rabbits, respectively. All dermal irritation completely subsided by day 13.
Other effects:
There were no deaths during the study.
There were no remarkable body weight changes during the study period.

Table 1. Individual irritation scores (erythema)

ANIMAL

SEX

SITE

1H

24H

48H

72H

4D

5D

6D

7D

8D

9D

10D

11D

12D

13D

28438

M

A

1s

1s

2s

2

2

2d

2d

2d

1d

1d

1d

0d

-

-

28444

M

A

0s

0s

1s

2

2

2

1d

1d

0d

-

-

-

-

-

28465

M

A

0s

1s

1

0

-

-

-

-

-

-

-

-

-

-

27714

F

A

0s

1s

3

3

3d

3d

2d

2d

2d

1d

1d

1d

1d

0d

28448

F

A

1s

1s

2s

3

3

2d

2d

2d

2d

1d

1d

1d

-

-

28458

F

A

0s

1s

2s

2

2

2

1d

1d

0d

-

-

-

-

-

Total

2

5

11

12

12

11

8

8

5

3

3

1

1

0

H = hours; D = days; F = female; M = male; - = dermal irritation previously subsided; d = desquamation; s = brown staining on dose site

Table 2. Individual irritation scores (oedema)

SITE

SEX

SITE

1H

24H

48H

72H

4D

5D

6D

7D

8D

9D

10D

11D

12D

13D

A

M

A

0

0

0

0

0

0

0

0

0

0

0

0

0

0

A

M

A

0

0

0

0

0

0

0

0

0

-

-

-

-

-

A

M

A

0

0

0

0

-

-

-

-

-

-

-

-

-

-

A

F

A

0

0

1

1

1

1

0

0

0

0

0

0

0

0

A

F

A

0

0

0

0

0

0

0

0

0

0

0

0

-

-

A

F

A

0

0

0

0

0

0

0

0

0

-

-

-

-

-

Total

 

 

0

0

1

1

1

1

0

0

0

0

0

0

-

-

Primary Irritation Index (PII) was calculated using test periods: 1h, 24h, 48h, and 72h

PII = [(Total erythema scores (2 + 5 + 11 + 12)) / 24 (amount of test sites)] + [(Total oedema scores (0 + 0 + 1 + 1)) / 24 (amount of test sites)] = 1.4 (slightly irritating)

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
According to the classification criteria outlined in the section 3.2.2.3.2.2. (Guidance on the Application of CLP criteria, 2013), if the mean value of ≥ 2.3 - ≤ 4.0 was reached for erythema/eschar or for oedema in at least 4 of 6 tested animals from gradings at 24, 48 and 72 hours after patch removal, the classification as Skin Irritant would be assigned. The evaluation of test results was performed according to the example 3 (test carried out with more than three animals; section 3.2.2.3.2.2.). Only 1/6 of the animals reached the cut-off value of 2.3 for erythema, i.e. only animal No. 27714 (female) is a positive responder. All other mean scores for erythema and oedema were under the cut-off value of 2.3. The dermal reactions were completely reversible within 13 days. Based on this knowledge, the target substance (Benzenesulfonic acid, mono-C20-24 (even)-sec-alkyl derivs., para-, sodium salts) does not meet criteria of Regulation (EC) No. 1272/2008 for classification and labelling as a skin irritant.
Executive summary:

The primary dermal irritation potential of C20-24 sodium sulfonate (TNB=448) was evaluated in this study with New Zealand White rabbits. There was one group of six albino rabbits that received a single, four-hour, semioccluded exposure. Each 0.5 mL dose of the test article was applied to the clipped, unabraded skin. At completion of exposure, the bandages were removed and the sites washed. Application sites were evaluated in accordance with the method of Draize at approximately 30-60 minutes and 24, 48 and 72 hours after patch removal and daily through day 13, if irritation persisted. There were no deaths or remarkable body weight changes during the study period. The test article induced very slight to moderate erythema on all animals and very slight oedema for one rabbit. Desquamation and brown staining of the skin were noted for five and six rabbits, respectively. All dermal irritation completely subsided by day 13. The Primary Irritation Index was calculated to be 1.4. The test article, C20 -24 sodium sulfonate (TBN=448), received a descriptive rating classification of slightly irritating.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation, other
Remarks:
in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1999-09-23 to 1999-10-18
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Version / remarks:
the European Economic Communities (EEC) Guideline in the Official Journal of the European Communities [92/69, Annex V, B5 (1992)l
Deviations:
no
Qualifier:
according to
Guideline:
other: the Japanese Agricultural Chemicals Laws and Regulations Testing Guidelines for Toxicology Studies published by the Society of Agricultural Chemicals Industry, under the auspices of the (MAFF) [59 NohSan No. 4200 (1985)
Deviations:
no
GLP compliance:
yes
Remarks:
in compliance with the United States Environmental Protection Agency (EPA) Good Laboratory Practice Regulations (40 CFR Parts 160 and 792)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Covance Research Products, Inc. Denver, PA
- Age at study initiation: Young adult
- Weight at study initiation: 3295 to 3615 grams at initiation of dosing
- Housing: Individually in suspended wire mesh cages
- Diet: PMI Nutrition International, Inc. Certified Rabbit LabDiet@ 5322 was offered at approximately 150 g per day during the study.
- Water (e.g. ad libitum): Municipal water was provided ad libitum.
- Acclimation period: for a minimum of seven days prior to initiation of dosing.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): (68.2 - 69.1°F; 20 - 21 °C)
- Humidity (%): 30.2-64.1
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
Prior to use, the original container of the test article was agitated to insure a homogeneous mixture. A sufficient amount of test article was dispensed into a labeled storage container and a stir bar was added. The test article was stirred continuously throughout use.
The test article was placed directly into the cupped lower conjunctival sac of each rabbit's right (test) eye. The eyelids were held closed for approximately one second after instillation. Left eyes were manipulated in an identical manner to simulate the dosing of the right eyes.
- Amount(s) applied (volume or weight with unit): 0.1 mL
Duration of treatment / exposure:
single, unwashed exposure.
Observation period (in vivo):
at approximately one, 24, 48 and 72 hours after dosing and on days 4, 7, 14 and 21, if irritation persisted.
Number of animals or in vitro replicates:
6 (Two males and four females)
Details on study design:
REMOVAL OF TEST SUBSTANCE: Washing: no
SCORING SYSTEM: presented in "Any other information on materials and methods incl. tables".
TOOL USED TO ASSESS SCORE: fluorescein, ophtalmoscope
Irritation parameter:
maximum mean total score (MMTS)
Basis:
mean
Time point:
other: 24, 48 and 72 hour
Score:
12.2
Max. score:
110
Reversibility:
fully reversible within: 21 days
Remarks on result:
other: at one-hour post-instillation
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 24, 48 and 72 hours
Score:
0
Max. score:
0
Reversibility:
other: not applicable
Remarks on result:
other: no findings
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 24, 48 and 72 hours
Score:
0
Max. score:
0
Reversibility:
other: not applicable
Remarks on result:
other: no findings
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #1
Time point:
other: mean of 24, 48 and 72-h readings
Score:
2
Max. score:
2
Reversibility:
fully reversible within: 14 days
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #2
Time point:
other: mean of 24, 48 and 72-h readings
Score:
2
Max. score:
2
Reversibility:
fully reversible within: 21 days
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #3
Time point:
other: mean of 24, 48 and 72-h readings
Score:
1.7
Max. score:
2
Reversibility:
fully reversible within: 4 days
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #4
Time point:
other: mean of 24, 48 and 72-h readings
Score:
2
Max. score:
2
Reversibility:
fully reversible within: 14 days
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #5
Time point:
other: mean of 24, 48 and 72-h readings
Score:
2
Max. score:
2
Reversibility:
fully reversible within: 7 days
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #6
Time point:
other: mean of 24, 48 and 72-h readings
Score:
2
Max. score:
2
Reversibility:
fully reversible within: 14 days
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: mean of 24, 48 and 72-h readings
Score:
0.7
Max. score:
1
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
other: mean of 24, 48 and 72-h readings
Score:
1
Max. score:
2
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
other: mean of 24, 48 and 72-h readings
Score:
0.7
Max. score:
1
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
chemosis score
Basis:
animal #4
Time point:
other: mean of 24, 48 and 72-h readings
Score:
0.7
Max. score:
2
Reversibility:
fully reversible within: 48 hours
Irritation parameter:
chemosis score
Basis:
animal #5
Time point:
other: mean of 24, 48 and 72-h readings
Score:
1
Max. score:
2
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
chemosis score
Basis:
animal #6
Time point:
other: mean of 24, 48 and 72-h readings
Score:
1
Max. score:
2
Reversibility:
fully reversible within: 72 hours
Irritant / corrosive response data:
No animals vocalized upon instillation of the test article. All animals were noted with conjunctival irritation. Iridal irritation was noted for one animal. There were no corneal findings. All irritation completely subsided by day 21. The left (control) eyes were free of evidence of ocular irritation and other findings for the duration of the study.
Other effects:
There were no deaths or remarkable body weight changes during the study period.

Table 1. Individual ocular irritation scores

 

 

 

Examination intervals

 

Animal

Sex

Tissue

1h

24h

48h

72h*

4 days

7 days*

14 days*

21 days*

Mean (24-48-72 h)

29139

M

Cornea (O-A)

0-0

0-0

0-0

0-0

0-0

0-0

0-0

-

0.0

 

 

Iris

1

0

0

0

0

0

0

-

0.0

 

 

Conjunctiva (R-C-D)

2-2- 2

2-1-2

2-1-0

2-0-0

2-0-0

2-0-0

0-0-0

-

2.0 (R) - 0.7 (C)

29143

M

Cornea (O-A)

0-0

0-0

0-0

0-0

0-0

0-0

0-0

0-0

0.0

 

 

Iris

0

0

0

0

0

0

0

0

0.0

 

 

Conjunctiva (R-C-D)

2-2-2

2-2-0

2-1-0

2-0-0

2-0-0

2-0-0

1-0-0

0-0-0

2.0 (R) – 1.0 (C)

29158

M

Cornea (O-A)

0-0

0-0

0-0

0-0

0-0

-

-

-

0.0

 

 

Iris

0

0

0

0

0

-

-

-

0.0

 

 

Conjunctiva (R-C-D)

2-1-1

2-1-0

2-1-0

1-0-0

0-0-0

-

-

-

1.7 (R) - 0.7 (C)

29148

F

Cornea (O-A)

0-0

0-0

0-0

0-0

0-0

0-0

0-0

-

0.0

 

 

Iris

0

0

0

0

0

0

0

-

0.0

 

 

Conjunctiva (R-C-D)

2-2-2

2-2-0

2-0-0

2-0-0

2-0-0

1-0-0

0-0-0

-

2.0 (R) - 0.7 (C)

29155

F

Cornea (O-A)

0-0

0-0

0-0

0-0

0-0

0-0

-

-

0.0

 

 

Iris

0

0

0

0

0

0

-

-

0.0

 

 

Conjunctiva (R-C-D)

2-2-2

2-2-0

2-1-0

2-0-0

2-0-0

0-0-0

-

-

2.0 (R) – 1.0 (C)

29156

F

Cornea (O-A)

0-0

0-0

0-0

0-0

0-0

0-0

0-0

-

0.0

 

 

Iris

0

0

0

0

0

0

0

-

0.0

 

 

Conjunctiva (R-C-D)

2-2-2

2-2-0

2-1-0

2-0-0

2-0-0

1-0-0

0-0-0

-

2.0 (R) – 1.0 (C)

Total

 

 

73

44

34

22

20

12

2.0

0.0

 

Mean

 

 

12.2

7.3

5.7

3.7

3.3

2.0

0.3

0.0

 

Mean group scores

 

Cornea

 

 

 

 

 

 

 

 

0.0

 

 

Iris

 

 

 

 

 

 

 

 

0.0

 

 

Conjunctiva

 

 

 

 

 

 

 

 

2.0 (R) – 0.9 (C)

*Sodium fluorescein in solution applied;

O = opacity;
A = area;

R = redness;

C = chemosis;

D = discharge;

h = hours;

M = male;

F = female

- = no observation made anymore 

Interpretation of results:
irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
According to the classification criteria outlined in the section 3.3.2.3.2.2. (Guidance on the Application of CLP criteria, 2013), if a substance cause positive but reversible reactions such as corneal opacity (score ≥ 1), iritis (score ≥ 1), conjunctival redness (score ≥ 2) or conjunctival oedema (chemosis) (score ≥ 2) in at least 4 of 6 tested animals from gradings at 24, 48 and 72 hours after application, the classification as Eye Irritant (Category 2) is assigned. The average irritation scores over 24, 48, and 72 hours for both corneal opacity and iritis were 0.0 in each of six animals. The mean conjunctival chemosis scores (swelling) for each animal were all under the cut-off value of 2.0 for this time period. However, the positive responses (score of 2.0) were observed in 5 of 6 animals regarding conjunctival erythema (redness). The irritation reactions were completely reversible within 21 days. Based on this observation, the target substance (Benzenesulfonic acid, mono-C20-24 (even)-sec-alkyl derivs., para-, sodium salts) is classified as Eye Irritant (Category 2, H319) according to Regulation (EC) No. 1272/2008.
Executive summary:

The primary ocular irritation potential of C20 -24 sodium sulfonate (TBN=448) was evaluated in this study with New Zealand White rabbits. There was one group of six albino rabbits that received a single, unwashed exposure. Each 0.1 mL dose of the test article was instilled into the lower conjunctival sac of the right eye. The eyelid was held closed for approximately one second and released. The left eye was manipulated in a similar manner as the right eye and served as a contralateral control. The eyes were examined for ocular reactions in accordance with the method of Draize at approximately one, 24, 48 and 72 hours after dosing and on days 4, 7, 14 and 21, if irritation persisted. Sodium fluorescein was used to aid in revealing possible corneal damage at 72 hours and on days 7, 14 and 21. There were no deaths or remarkable body weight changes during the study period. All animals were noted with positive (grade 2) conjunctival irritation. Iridal irritation was noted for one animal. There were no corneal findings. All irritation completely subsided by day 21. The Maximum Average Score for C20 -24 sodium sulfonate (TBN=448) was 12.2 at one-hour post-instillation.

The following table presents the mean scores calculated from individual scores:

Conjunctiva

-      Redness

-      Oedema

24 and 72 hours

24,48 and 72 hours

1.9

2.0

0.8

0.9

Iris

0.0

0.0

Cornea

0.0

0.0

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Additional information

Skin irritation

In the key study, the primary dermal irritation potential of Benzenesulfonic acid, mono-C20-24 (even)-sec-alkyl derivs., para-, sodium salts (generic name: C20-24 sodium sulfonate) containing para dodecylphenol (PDDP, CAS 210555-94-5) as impurity was evaluated in a study with New Zealand White rabbits (Kern, 1999a; Report No. WIL-168153). There was one group of six albino rabbits that received a single, four-hour, semioccluded exposure. Each 0.5-mL dose of the test article was applied to the clipped, unabraded skin. At completion of exposure, the bandages were removed and the sites washed. Application sites were evaluated in accordance with the method of Draize at approximately 30-60 minutes and 24, 48 and 72 hours after patch removal and daily through day 13, if irritation persisted. There were no deaths or remarkable body weight changes during the study period. The test article induced very slight to moderate erythema on all animals and very slight edema for one rabbit. Desquamation and brown staining of the skin were noted for five and six rabbits, respectively. All dermal irritation completely subsided by day 13. The Primary Irritation Index was calculated to be 1.4. The test article received a descriptive rating classification of slightly irritating.

In several supporting studies, the read-across substances calcium sulfonates (CAS 70024-69-0 and CAS 61789-86-4) were not irritating to rabbit' skin (Kern, 1999b, c; Swan, 1972; Hoff, 2002a, b; Buehler, 1990a, 1991b; Costello, 1986b; Ohees, 1968c; Gabriel, 1981b).

In a supporting study (Kern, 1999b, according to OECD 404) skin irritation potential of the calcium sulfonate read across substance (CAS 70024-69-0) was investigated in New Zealand White rabbits. 3 males and 3 females were treated semi-occlusively (type of cover: gauze binder) with 0.5 mL of the test material for 4 hours. After removal of the substance the animals were observed for 8 days. The test sites were then evaluated in accordance with the method of Draize at 60 min, 24, 48 and 72 hours and then daily if irritation persisted. The mean erythema score were 1, 1, 1, 1.7, 1 and 1 and were fully reversible by maximally 8 days. Mean oedema scores at 24, 48 and 72 hours were 1, 0, 0.3, 0, 0.3 and 0, and were fully reversible within 48 h or 4 days. Animals did not display any abnormal signs throughout the test period. Bodyweights did not significantly alter. Based on the results of this study this substance is not a skin irritant.

In another supporting primary dermal irritation study the calcium sulfonate read across substance (CAS 70024-69-0; Swan, 1972) was evaluated in albino rabbits. In this study, a dose of 0.5 mL of the undiluted test substance was applied to the abraded an unabraded skin of 6 young adult albino rabbits for 24 hours. Animals then were observed at the end of the 24 hour exposure and at 72 hours. All scores for erythema and edema in abraded and unabraded skin at both the 24 hour and 48 hour observations were 0 using the method of Draize. Based on the absence of effects after 24 hour exposure, which is longer (more severe) than the current prescribed 4 hour exposure guidance, this substance is not a skin irritant.

In another supporting study with the calcium sulfonate read across substance CAS 70024-69-0 (Hoff, 2002a), New Zealand White rabbits (3 animals) were treated semi-occlusively (type of cover: plastic) with 0.5 mL of the test material for 4 hours. After removal of the substance (gently washing with distilled water) the animals were observed for 14 days. The test sites were then evaluated in accordance with the method of Draize at 24, 48 and 72 hours and then daily if irritation persisted. The mean erythema scores were 0.7, 0 and 1.7, and were fully reversible by 72 hours or 14 days for 1.7. Mean oedema scores at 24, 48 and 72 hours were 0, 0 and 1, and were fully reversible within 14 days. Animals did not display any abnormal signs throughout the test period. Bodyweights did not significantly alter. Based on the results of this study this substance is not a skin irritant.

In another supporting study, the dermal irritation potential of the same calcium sulfonate read across substance (CAS 70024-69-0) was tested (Hoff, 2002b). In this study, 3 young adult New Zealand White rabbits were dermally exposed (semi-occlusively) to 0.5 mL of the test item for 4 hours. After removal of the substance (gently mashing with distilled water), animals then were observed for 7 days. The test sites were then evaluated in accordance with the method of Draize at 24, 48 and 72 hours and then daily if irritation persisted. The mean erythema score were 1, 1 and 2, and were fully reversible within 7 days. Mean oedema scores at 24, 48 and 72 hours were 0, 0 and 1.3, and were fully reversible within 7 days. Animals did not display any abnormal signs throughout the test period. Bodyweight was reduced in one animal. Mean scores for each animal at 24, 48 and 72 hours for erythema and oedema were less than 2.3 and fully reversible by maximally 7 days, respectively. Based on the results of this study this substance is not a skin irritant.

In a disregarded skin irritation screening study (Buehler, 1990a) the calcium sulfonate read across substance, CAS 70024-69-0 was investigated in 2 rabbits (1 male and 1 female). A dose of 0.5 mL of the undiluted test material was placed on gauze, which was applied to the shaved skin and then covered with a semi-occlusive wrap. After 4 hours the wrap and gauze were removed and the skin of each animal was evaluated at 24 and 72 hours in accordance with the method of Draize. The mean erythema score were 2.5 in one animal and 2.0 in the other. In the former animal erythema decreased from a score of 3 at 24 hours to a score of 2 at 72 hours indicating the effects are reversible. While the overall mean scores for erythema and oedema (2.5 and 2.25, respectively, with evidence of reversibility) suggest that the substance is not a skin irritant, this study cannot be used to determine classification and labelling because only 2 animals were used and observations were terminated at 72 hours rather than the normal 14 days.

In another disregarded study of the calcium sulfonate read across substance, CAS 70024-69-0 that was conducted following the disregarded screening study of Buehler (1990a), six animals (3 male and 3 female animals) were used to more fully evaluate the skin irritation potential of the calcium sulfonate read across substance, CAS 70024-69-0 (Buehler, 1991a). A dose of 0.5 mL of four different concentrations (6.25 %, 12.5 %, 25 % and 50%). the test material was placed on gauze, which was applied to the shaved skin and then covered with a semi-occlusive wrap. After 4 hours the wrap and gauze were removed and the skin of each animal was evaluated at 24 and 72 hours in accordance with the method of Draize. The mean erythema score for 6.25 % were 1.5, 2, 2, 2, 2 and 1, and were not fully reversible within 72 hours. Mean oedema scores for 6.25 % at 24 and 72 hours were 0.5, 0.5, 0, 2, 0.5 and 1, and were mainly not fully reversible within 72 hours. The mean erythema score for 12.5 % were 1.5, 2, 2, 2, 2.5 and 2, and were not fully reversible within 72 hours. Mean oedema scores for 6.25 % at 24 and 72 hours were 0.5, 0.5, 0.5, 1, 1.5 and 0.5, and were mainly not fully reversible within 72 hours. The mean erythema score for 25 % were 2, 3, 3, 2.5, 2 and 2, and were not fully reversible within 72 hours. Mean oedema scores for 25 % at 24 and 72 hours were 2, 2.5, 1.5, 2.5, 0.5 and 2, and were mainly not fully reversible within 72 hours. Mean scores for each animal at 24 h, and 72 h for erythema and oedema (concentration ≤ 25%), respectively, were less than 2.3.The mean erythema score for 50 % at 24 and 72 hours were 2, 3, 3, 3, 2.5 and 1.5, and were not fully reversible within 72 hours. Mean oedema scores for 50 % at 24 and 72 hours were 2.5, 3, 2.5, 2.5, 2 and 0.5, and were mainly not fully reversible within 72 hours. However, the skin irritation response in this study, particularly at 50%, is disregarded because it is inconsistent with the skin irritation results from the key and supporting studies on the calcium sulfonate read across substances (CAS 70024-69-0 and CAS 61789-86-4.

In another supporting primary dermal irritation study, 6 albino rabbits were dermally exposed to the test article (as supplied, dose volume 0.5 mL) the calcium sulfonate read across substance (CAS 61789 -86 -4) for 4 hours (Costello, 1986b). The skin was clipped over a wide area on their backs approximately 24 hours prior to application. Animals then were observed for 72 hours. Irritation was scored by the method of Draize. The mean 24, 48, and 72 hour individual animal erythema and oedema scores were 1.33, 1.67, 1.0, 1.67, 1.67, and 1.67 and 0.33, 1.67, 1.33, 2.0, 2.33, and 2.0, respectively. The overall mean 24, 48, and 72 hour erythema and edema scores were 1.5 and 1.6, respectively. Scores of 1 for erythema and oedema were present in one or more animals at the end of the 14 day observation period. Based on the results of this study this substance is not a skin irritant.

In another supporting primary dermal irritation study, 6 albino rabbits were dermally exposed to the test article (as supplied, dose volume 0.5 mL) the calcium sulfonate read across substance (CAS 61789 -86 -4) for 24 hours (Ohees, 1968c). The test material was applied to two areas, one abraded area and one intact area. The animals were fitted with collars for a 24 hour period at which time the coverings were removed, the test material washed off and the degree of erythema and oedema were recorded. A second reading was taken at 72 hours. The mean 24 and 72 hour individual animal erythema and oedema scores for abraded skin were 0.5, 0.5, 0.5, 0.5, 0.5, and 0.5 and 0.0, 0.5, 0.0, 0.5, 0.5, and 0.0, respectively. The mean 24 and 72 hour individual animal erythema and oedema scores for unabraded skin were 0.5, 0.5, 0.5, 0.5, 0.5, and 0.5 and 1.67 and 0.0, 0.5, 0.0, 0.5, 0.5, and 0.0, respectively. The overall mean 24 and 72 hour erythema and edema scores for abraded and unabraded skin were 0.5 and 0.25, respectively. All erythema and oedema scores in abraded and unabraded skin were 0.0 at the final 72 hour observation period. Based on the results of this study this substance is not a skin irritant.

In another disregarded primary dermal irritation study, 6 albino rabbits were dermally exposed to the sulfonate read across substance (CAS 61789 -86 -4, as supplied, dose 0.5 g) for 24 hours (Gabriel, 1981b). The test material was applied to two areas, one abraded area and one intact area. Gauze patches were placed over the treated area and an impervious material was wrapped snugly around the trunks of the animals to hold the patches in place. The skin of each animal was evaluated at 24 and 72 hours in accordance with the method of Draize. The mean erythema scores were 2 (intact skin) and 2 (abraded skin) after 24 hours and 1.83 (intact skin) and 1.67 (abraded skin) after 72 hours. Mean oedema scores at 24 and 72 hours were 1.5 and 0.67 referring to intact skin. In addition, scores of 1.5 and 0.67 were determined for abraded skin. However, this study cannot be used to determine classification and labelling because the exposure period was 24 hours rather than 4 hours and the study was terminated prior to 14 days post exposure. The observed skin irritation scores following 24 hours cannot be used to predict the scores following the shorter (less severe) 4 hour exposure prescribed under current guidance. 

The primary dermal irritation potential of the calcium sulfonate read across substance (CAS 61789-86-4) was evaluated in this study with New Zealand White rabbits (Kern, 1999c). There was one group of six albino rabbits that received a single, four-hour, semioccluded exposure. Each 0.5 mL dose of the test article was applied to the clipped, unabraded skin. At completion of exposure, the bandages were removed and the sites washed. Application sites were evaluated in accordance with the method of Draize at approximately 30-60 minutes and 24, 48 and 72 hours after patch removal and daily through day 13, if irritation persisted. The mean 24, 48, and 72 hour individual animal erythema and oedema scores were 1.0, 1.33, 0.33, 0.0, 0.0, and 0.67 and 0.0, 0.33, 0.0, 0.0, 0.0, and 0.5, respectively. The overall mean 24, 48, and 72 hour erythema and oedema scores were 0.5 and 0.05, respectively. All erythema and oedema was fully reversible by day 13 and 72 hours, respectively. Based on the results of this study this substance is not a skin irritant.

Based on these results, the calcium sulfonates do not meet criteria for classification and labelling as skin irritants in accordance with EU CLP Regulation (EC) No. 1272/2008.

The petroleum derived sodium sulfonates (CAS 68608 -26 -4) were highly irritating and corrosive in two skin irritation studies in rabbits, respectively (Gilman, 1982; Costello, 1983). Such a result is probably due to the exposure time of 24 hours. Therefore, these studies have been disregarded.

Eye irritation

The primary ocular irritation potential of Benzenesulfonic acid, mono-C20-24 (even)-sec-alkyl derivs., para-, sodium salts (generic name: C20 -24 sodium sulfonate) containing para dodecylphenol (PDDP, CAS 2105555 -94 -5) as impurity was evaluated in a study with New Zealand White rabbits (Kern, 1999d, Report No. WIL-168171). There was one group of six albino rabbits that received a single, unwashed exposure. Each 0.1-mL dose of the test article was instilled into the lower conjunctival sac of the right eye. The eyelid was held closed for approximately one second and released. The left eye was manipulated in a similar manner as the right eye and served as a contralateral control. The eyes were examined for ocular reactions in accordance with the method of Draize at approximately one, 24, 48 and 72 hours after dosing and on days 4, 7, 14 and 21, if irritation persisted. Sodium fluorescein was used to aid in revealing possible corneal damage at 72 hours and on days 7, 14 and 21. There were no deaths or remarkable body weight changes during the study period. All animals were noted with positive (grade 2) conjunctival irritation. Iridal irritation was noted for one animal. There were no corneal findings. All irritation completely subsided by day 21. The Maximum Average Score for the target substance was 12.2 at one-hour post-instillation. The mean group scores were 0.0, 0.0, 2.0 and 0.9 for cornea, iris, conjunctival redness and chemosis, respectively.

In several disregarded studies, the read-across substances calcium sulfonates (CAS 70024-69-0 and CAS 61789-86-4) were not irritating to eyes of rabbits (Kern, 1999e; Swan, 1972; Buehler, 1991b; Ohees, 1968d; Gabriel, 1981c), although one study with CAS 70024-69-0 was positive (Buehler, 1990b; the study has deficiencies: it was terminated at 72 -hour observation time point).

In a disregarded study, the eye irritation potential of the calcium sulfonate read across substance, (CAS 70024-69-0), was evaluated in accordance withEPA OPPTS 870.2400 (Kern, 1999e). A group of six albino rabbits (3 per sex) was used for the study. Each animal received a single, unwashed exposure of 0.1 mL of the test article instilled into the lower conjunctival sac of the right eye. The eyelid was held closed for approximately one second and released. The left eye was manipulated in a similar manner as the right eye and served as a contralateral control. Observations for eye irritation were made at 1, 24, 48 and 72 hours following dosing and on day 4 in accordance with the method of Draize.One animal had clear discharge from the eye, one hour post dosing. There were no deaths or changes in bodyweight throughout the study.Mean 24, 48, and 72 hour corneal and iris scores were 0 for all animals tested. The conjunctival erythema score were 1.3, 1, 1, 1, 1, and 1,respectively andwere all reversible within 7 days. The chemosis scores were 0.3, 1.3, 0.3, 0, 0 and 1, respectively.In this study, substance is not an eye irritant.

In another disregarded study, the eye irritation potential of the calcium sulfonate read across substance, (CAS 70024-69-0), was evaluated (Swan, 1972). Adult New Zealand white rabbits (3/sex) were administered 0.1 mL of test substance into one eye. The other eye served as a control. The test material was not washed from the eyes. Eyes were scored for corneal, iritis and conjunctival effects 24, 48 and 72 hours following exposure in accordance with the method of Draize.Mean scores at 24, 48 and 72 hours for each animal for corneal opacity and iritis was 0 for all animals. Mean scores for conjunctival redness were 2, 2, 1.3, 1.3, 1 and 1, respectively. Mean scores for chemosis were 2, 1.7, 0.7, 1, 0.3 and 1, respectively. As the scores for conjunctival redness and oedema were less than 2 in 4 animals, therefore, this substance is not irritating to rabbit eyes.

Moreover, data on calcium sulfonate read-across substance, (CAS 70024-69-0), in rabbits show that this substance is not irritating to the eyes.

In another disregarded eye irritation screening study (Buehler, 1990b) the calcium sulfonate read across substance, CAS 70024-69-0 was investigated in 2 rabbits (1 male and 1 female). Each animal received a single, exposure of 0.1 mL of the test article instilled into one eye. The other eye served as a control. The test material was not washed from the eyes. Animals were observed at 1, 24, 48, 72 hours in accordance with the method of Draize. Mean scores at 24, 48 and 72 hours for each animal for corneal opacity and iritis were 0 for all animals. Mean scores for conjunctival redness was 2 in both animals. Conjunctival oedema scores were 2.0 in one animal and 2.33 for the other. In the latter animal the conjunctival oedema decreased from a score of 3 at 24 hours to a score of 2 at 72 hours indicating the effects being reversible. However, because only 2 animals were used and observations were terminated at 72 hours rather than the normal 21 days, this study cannot be used to determine classification and labelling.

In a follow-up disregarded study of the calcium sulfonate read across substance, CAS 70024-69-0, that was conducted following the screening study of Buehler (1991b), six animals (3 male and 3 female animals) were used to more fully evaluate the eye irritation potential of the calcium sulfonate read across substance, CAS 70024-69-0. Each animal received a single, exposure of 0.1 mL of the test article instilled into one eye. The other eye served as a control. The test material was not washed from the eyes. Animals were observed at 1, 24, 48, 72 hours in accordance with the method of Draize. Mean scores at 24, 48 and 72 hours for each animal for corneal opacity and iritis was 0 for all animals. Mean scores for conjunctival redness were 1, 0.7, 0.7, 0.3, 0.3 and 1, respectively. Mean scores for chemosis were 1.3, 0.7, 0, 0.7, 0.7 and 1.7, respectively. With the exception of the 1.7 value for chemosis in animal 6, all other signs were fully reversible within 72 hours. The mean scores for all animals for conjunctival redness and oedema were less than 2. Therefore, this substance is not irritating to rabbit eyes.

In another disregarded study the calcium sulfonate read across substance (CAS 61789-86-4) was investigated for its eye irritating potential (Ohees, 1968d).Adult albino rabbits of the New Zealand White variety (6) were placed in a collar such that the animals could not rub their eyes. One tenth of a millilitre of test substance was instilled in one eye, the other untreated eye served as a control. The reaction to the test material was read according to the scale of scoring for damage to the cornea, iris and the bulbar and palpebral conjunctivae at 24, 48 and 72 hours after eye instillation. Any residue of the test material and accumulated discharge are flushed from the eye each time they are scored.

Under the conditions specified the product has an eye irritating score of 0 and 0 and 0 at 24, 48 and 72 hours respectively. Therefore the substance is not irritating to the eye.

In another disregarded study with the same calcium sulfonate read across substance, CAS 61789-86-4 (Gabriel, 1981c) was investigated for its eye irritating potential in rabbits. Six healthy young adult albino rabbits were used in this study. 0.1 gram of the experimental material was instilled into the right eyes of the test animals, while the other eye remained untreated to serve as a control. The test material was not washed from the eyes. The treated eyes were examined at one, two, three, five and seven days following instillation of the test material into the eyes. Interpretation of the results was made in accordance with the Draize Scale of Scoring Ocular Lesions. The test material is not a primary irritant and requires no cautionary labelling with respect to an eye irritating potential.

Based on these results the calcium sulfonate read-across substances do not meet criteria for classification and labelling as eye irritating in accordance with EU CLP Regulation (EC) No. 1272/2008.

Two eye irritation studies were conducted with a petroleum derived sodium salt (CAS 68608 -26 -4) (Costello, 1982, 1983). These studies also determined that sodium sulfonate is an eye irritant. However, these studies are disregarded for purposes of determining the classification and labelling of the C20-24 sodium sulfonate [TBN = 448] because the test materials were low TBN substance [TBN = 0]. Furthermore, the test material used in the study by Costello, 1982 is not typical of current production.

Results of the eye irritation studies with calcium sulfonates (CAS 70024-69-0 and CAS 61789 -86-4: Kern, 1999; Swan, 1972; Buehler, 1991; Ohees, 1968; Gabriel, 1981; Buehler, 1990) also are disregarded because they are not consistent with the results of eye irritation with sodium sulfonate or have deficiencies


Justification for selection of skin irritation / corrosion endpoint:
Only one study available

Justification for selection of eye irritation endpoint:
Only one study available

Effects on eye irritation: slightly irritating

Justification for classification or non-classification

The classification and labelling of the target substance Benzenesulfonic acid, mono-C20-24 (even)-sec-alkyl derivs., para-, sodium salts (generic name: C20 -24 sodium sulfonate) is based on the results of the key studies for this substance, but the results of the sodium and calcium sulfonate read-across substances have also been taken into account.

Skin irritation

According to the classification criteria outlined in the section 3.2.2.3.2.2. (Guidance on the Application of CLP criteria, 2013), if the mean value of2.3 -4.0 was reached for erythema/eschar or for oedema in at least 4 of 6 tested animals from gradings at 24, 48 and 72 hours after patch removal, the classification as Skin Irritant would be assigned. The evaluation of test results was performed according to the example 3 (test carried out with more than three animals; section 3.2.2.3.2.2.). Only 1/6 of the animals reached the cut-off value of 2.3 for erythema, i.e. only animal No. 27714 (female) is a positive responder. All other mean scores for erythema and oedema were under the cut-off value of 2.3. The dermal reactions were completely reversible within 13 days. Based on this knowledge, the C20-24 sodium sulfonate (Benzenesulfonic acid, mono-C20-24 (even)-sec-alkyl derivs., para-, sodium salts) does not meet criteria of Regulation (EC) No. 1272/2008 for classification and labelling as a skin irritant.

Eye irritation

According to the classification criteria outlined in the section 3.3.2.3.2.2. (Guidance on the Application of CLP criteria, 2013), if a substance cause positive but reversible reactions such as corneal opacity (score ≥ 1), iritis (score ≥ 1), conjunctival redness (score ≥ 2) or conjunctival oedema (chemosis) (score ≥ 2) in at least 4 of 6 tested animals from gradings at 24, 48 and 72 hours after application, the classification as Eye Irritant (Category 2) is assigned. The average irritation scores over 24, 48, and 72 hours for both corneal opacity and iritis were 0.0 in each of six animals. The mean conjunctival chemosis scores (swelling) for each animal were all under the cut-off value of 2.0 for this time period. However, the positive responses (score of 2.0) were observed in 5 of 6 animals regarding conjunctival erythema (redness). The irritation reactions were completely reversible within 21 days. Based on this observation, the C20-24 sodium sulfonate ( Benzenesulfonic acid, mono-C20-24 (even)-sec-alkyl derivs., para-, sodium salts) is classified as Eye Irritant (Category 2, H319) according to Regulation (EC) No. 1272/2008.