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Administrative data

Description of key information

Skin irritation, rabbits, OECD 404, not irritating to the skin
Skin irritation, rabbits, not irritating to the skin
Eye irritation, rabbits, OECD 405, not irritating to eyes
Eye irritation, rabbits, OECD 405, not irritating to eyes

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
21 Feb - 04 Mar 2008
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP - Guideline study with acceptable restrictions, in some cases the animals had already been used in earlier studies.
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
yes
Remarks:
in some cases the animals had already been used in earlier studies.
Qualifier:
according to guideline
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
yes
Remarks:
in some cases the animals had already been used in earlier studies.
GLP compliance:
yes (incl. QA statement)
Remarks:
Ministerium für Umwelt und Naturschutz, Landwirtschaft und Verbraucherschutz des Landes Nordrhein-Westfalen, Düsseldorf, Germany
Species:
rabbit
Strain:
other: Crl:KBL(NZW)BR
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River, Kißlegg, Germany
- Age at study initiation: young adult animals (females)
- Weight at study initiation: 3100 - 3400 g
- Housing: individual housing in Metall/Noryl (EBECO) cages, excrement trays below the cages contained low dust wood granulate bedding.
- Diet: standard diet "Ssniff K-Z" 4 mm (Spezialdiäten GmbH, Soest, Germany), approximately 100 g/animal/day. Hay or hay pellets were offered additionally (hay, irradiated, Harlan Winkelmann GmbH, Borchen Germany; hay pellets, Ssniff Spezialdiäten GmbH, Soest, Germany)
- Water: tap water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 50 ± 25
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
unchanged (no vehicle)
Controls:
other: untreated sites of the same animal served as control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL
- Concentration (if solution): 100%
Duration of treatment / exposure:
Initial test: 3 min, 1 and 4 h
Confirmatory test: 4 h
Observation period:
7 days
Reading time points: 1, 24, 48 and 72 h and 7 days
Number of animals:
3 females
Details on study design:
TEST SITE
- Area of exposure: 6.25 cm² on the dorso-lateral area of the trunk
- Type of wrap if used: The treated skin was covered with a gauze patch, held in place with non-irritating tape for the duration of the exposure period.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Residual test material was removed with water.
- Time after start of exposure: 3 min, 1 and 4 h

SCORING SYSTEM: Draize scoring system
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
other: mean over 24, 48 and 72 h
Score:
2.3
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: brownish discolouration of the treated skin was observed up to 1 h after patch removal
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
other: mean over 24, 48 and 72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: brownish discolouration of the treated skin was observed up to 1 h after patch removal
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
other: mean over 24, 48 and 72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
other: brownish discolouration of the treated skin was observed up to 24 h after patch removal
Irritation parameter:
edema score
Basis:
animal #1
Time point:
other: mean over 24, 48 and 72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
edema score
Basis:
animal #2
Time point:
other: mean over 24, 48 and 72 h
Score:
1.3
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
edema score
Basis:
animal #3
Time point:
other: mean over 24, 48 and 72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Other effects:
No other local or systemic effects were reported.

Table 1. Results of skin irritation study.  

Observation time

Rabbit no.

1

2

3

Erythema

Edema

Erythema

Edema

Erythema

Edema

1 h

2

0

1

0

1

0

24 h

3

3

2

2

2

1

48 h

2

2

2

1

2

1

72 h

2

1

2

1

2

1

7 d

0

0

0

0

0

0

Mean over 24 + 48 + 72 h

2.3

2.0

2.0

1.3

2.0

1.0

Mean over 24 + 48 + 72 h all animals

2.1

1.4

 

 

 

 

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
CLP: not classified
DSD: Xi, R38
Executive summary:

The study objective was to determine the irritant / corrosive effects on skin of female albino rabbits (strain Crl:KBL(NZW)BR). The method used complied with the OECD TG 404 and GLP, therefore the sudy is assigned as Klimish 1. 0.5 mL of the pure liquid test substance was applied to the skin of the animal under a gauze patch. The treated skin area was approximately 2.5 cm by 2.5 cm in size. The patch was placed on the dorso-lateral areas of the trunk of each animal and was held in place with non-irritating tape for the duration of the exposure period. After the exposure period the dressing and patch were removed. The exposed skin area was carefully washed with water without altering the existing response, or the integrity of the epidermis. The surrounding untreated skin served as control. In the first step only one animal was used and three test patches were applied successively to this animal. As no serious skin reactions were observed after three minutes, the second patch was applied and removed after one hour. At this stage the observations indicated that with respect to animal welfare the exposure can be allowed to extend to four hours, therefore the third patch was applied and removed after four hours and the responses were graded one hour later. The test was completed using two additional animals, exposed for four hours. The dermal irritation was scored approximately at 1, 24, 48 and 72 hours after patch removal. The dermal irritation was observed after 72 hours therefore the animals were monitored usually on day 7 and 14 after patch removal until the changes had completely subsided. The degree of erythema/eschar formation and oedema formation was recorded as specified by DRAIZE. Any serious lesions or toxic effects other than dermal irritation were also recorded and fully described. As general criteria the body weight of each animal was recorded at the beginning of the study. If clinical findings other than dermal irritations occur they were recorded daily. The mean irritation scores for erythema (redness) and eschar formation at the 24 -h, 48 -h and 72 -h reading points were 2.3, 2.0 and 2.0 for animal 1, 2 and 3, respectively. The mean oedema scores were 2.0, 1.3 and 1.0 for animal 1, 2 and 3, respectively. All skin reactions were fully reversible within 7 days. There were no systemic intolerance reactions. The irritation scores in 2 of 3 animals do not exceed the cut-off values defined in the CLP, therefore the classification as skin irritant is not warranted for this substance.

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

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
06 Mar - 18 Mar 2008
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP - Guideline study with acceptable restrictions, in some cases the animals had already been used in earlier studies.
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
in some cases the animals had already been used in earlier studies.
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
in some cases the animals had already been used in earlier studies.
GLP compliance:
yes (incl. QA statement)
Remarks:
Ministerium für Umwelt und Naturschutz, Landwirtschaft und Verbraucherschutz des Landes Nordrhein-Westfalen, Düsseldorf, Germany
Species:
rabbit
Strain:
other: Crl:KBL(NZW)BR
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Charles River, Kißlegg, Germany
- Age at study initiation: young adult animals (females)
- Weight at study initiation: 2300 - 2600 g
- Housing: individual housing in Metall/Noryl (EBECO) cages, excrement trays below the cages contained low dust wood granulate bedding.
- Diet: standard diet Ssniff (Spezialdiäten GmbH, Soest, Germany), approximately 100 g/animal/day. Hay or hay pellets were offered additionally (hay, irradiated, Harlan Winkelmann GmbH, Borchen Germany; hay pellets, Ssniff Spezialdiäten GmbH, Soest, Germany)
- Water: tap water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 50 ± 25
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
unchanged (no vehicle)
Controls:
other: the untreated eye served as control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
- Concentration (if solution): 100%
Duration of treatment / exposure:
24 h
Observation period (in vivo):
7 days
Reading time points: 1, 24, 48 and 72 h and 7 days
Number of animals or in vitro replicates:
3 females
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): the treated eye was rinsed
- Time after start of exposure: 24 h

SCORING SYSTEM: Draize scoring system

TOOL USED TO ASSESS SCORE: fluorescein
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
out of all 3 animals
Time point:
other: mean over 24, 48 and 72 h
Score:
0
Max. score:
4
Reversibility:
other: reversibility: not applicable
Irritation parameter:
iris score
Basis:
mean
Remarks:
out of all 3 animals
Time point:
other: mean over 24, 48 and 72 h
Score:
0
Max. score:
2
Reversibility:
other: reversibility: not applicable
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
other: mean over 24, 48 and 72 h
Score:
1.3
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
other: mean over 24, 48 and 72 h
Score:
1.3
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
other: mean over 24, 48 and 72 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 72 h
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: mean over 24, 48 and 72 h
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: 48 h
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
other: mean over 24, 48 and 72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
other: mean over 24, 48 and 72 h
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: 48 h
Irritant / corrosive response data:
Cornea/Iris: No effects on cornea or iris were observed during the study period in any animal.
Conjunctivae: At the 1 and 24 h reading time point all animals showed moderate conjunctivae redness. After 48 h mild redness was apparent in all animals, still persistent in 2/3 animals after 72 h. All conjunctivae redness was fully reversible within 72 h and 7 days, respectively.
Chemosis: 3/3 animals showed mild swelling after 24 h which was fully reversible in 2/3 animals after 48 h. In 1/3 animals chemosis was persistent up to 72 h and fully reversible after 7 days.
Other effects:
No further local or systemic effects were reported.

 Table 1. Results of eye irritation study. 

Rabbit #

Time [h]

conjunctivae

 

iris

cornea

redness

swelling

1

1

2

0

0

0

24

2

1

0

0

48

1

0

0

0

72

1

0

0

0

168

0

0

0

0

average

1.3

0.3

0.0

0.0

2

1

2

0

0

0

24

2

1

0

0

48

1

1

0

0

72

1

1

0

0

168

0

0

0

0

average

1.3

1.0

0.0

0.0

3

1

2

0

0

0

24

2

1

0

0

48

1

0

0

0

72

0

0

0

0

average

1.0

0.3

0.0

0.0

 

 

Time [h]

conjunctivae

 

iris

cornea

 

redness

swelling

 

 

mean over

1

2.00

0.00

0.00

0.00

24

2.00

1.00

0.00

0.00

48

1.00

0.33

0.00

0.00

72

0.67

0.33

0.00

0.00

24+48+72

1.22

0.56

0.00

0.00

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
CLP: not classified
DSD: not classified
Executive summary:

The study objective was to determine the irritant effects of the target substance on eyes of albino rabbits (strain Crl:KBL(NZW)BR) (Gmelin, 2008). The study was conducted according to OECD TG 405 and GLP and is assigned as Klimish 1. 0.1 mL of the pure liquid test substance was placed into the conjunctival sac of one eye of the first animal after having gently pulled the lower lid away from the eyeball. The lids were gently held together for about one second in order to prevent loss of the test compound. The other eye, which remained untreated, served as control. The eye was rinsed approximately 24 hours following instillation. There was no severe irritation observed in the first animal, therefore two further rabbits were treated in the same manner. Eye irritations were scored and recorded approximately at 1, 24, 48 and 72 hours post application. Irritation indices were observed after 72 h, therefore the animals were monitored on day 7, 14 and 21 after application until the changes had completely subsided. The degree of ocular lesions was recorded as specified by Draize. The body weight of each animal was recorded at the beginning of the study. If clinical findings other than eye irritations occur they were recorded daily.

The mean scores at 24 -h, 48 -h, and 72 -h reading time points were 0.0 for corneal opacity and iritis for three animals. Redness sconjunctivae scores were 1.3, 1.3 and 1.0 and chemosis conjunctivae scores were 0.3, 1.0 and 0.3 for animal 1, 2, and 3, respectively. All irritation reactions were fully reversible within 7 days. There were no systemic intolerance reactions. According to the classification criteria (CLP/GHS), the substance is not irritating to eyes.

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation:

According to the sequential testing strategy, a preliminary in vitro study for predicting non-specific, corrosive potentials of Benzenesulfonic acid, di-C10-14-alkyl derivs., calcium salts by using reconstructed human skin (RHS) was performed (Vohr, 2008). The experiment was carried out on a reconstructed human epidermis EST-1000 (CellSystems, St. Katharinen, Germany) for detection of topically applied skin corrosives with the test material. Corrosive skin effects of substances were defined as irreversible damage of the skin; namely, visible necrosis through the epidermis and into the dermis, following the application of a test substance for up to four hours. In vivo corrosive reactions are typified by ulcers, bleeding, and bloody scabs. A 100% concentration was tested on the skin/epidermal equivalents in triplets. For the determination of time related cytotoxic effects the incubation periods were 3 min and 60 min, respectively. By this the study was conducted in accordance with the OECD 431 guideline as well as with an EC draft guideline (amending Council Directive 67/548/EEC, B.40 Skin corrosion) using the test item concentrations and incubation periods recommended there. These tests are also related to the revised OECD 404 guideline "Acute Dermal Irritation/Corrosion". To check the reliability of this test procedure a blinded positive/negative control study is conducted at regular intervals in the lab. This reliability check is always performed by two technicians, two laboratories in parallel using the same batches of the control test items. The test item was applied at a 100% concentration, i.e. 50 µL per insert. The MTT (Methylthiazoletetrazolium) method has determined the following values of viability after 3 min. or after 60 min. of incubation: 101.50% and 99.56%, respectively. Thus, the results show that no corrosive property of the test item was determined by the assay used.

The key study was conducted with the target substance (Benzenesulfonic acid, di-C10-14-alkyl derivs., calcium salts) to determine the irritant effects on skin of female albino rabbits (strain Crl:KBL(NZW)BR) (Gmelin, 2008). The method used complied with the OECD TG 404 and GLP, therefore the study is assigned as Klimish 1. 0.5 mL of the pure liquid test substance was applied to the skin of the animal under a gauze patch. The treated skin area was approximately 2.5 cm by 2.5 cm in size. The patch was placed on the dorso-lateral areas of the trunk of each animal and was held in place with non-irritating tape for the duration of the exposure period. After the exposure period the dressing and patch were removed. The exposed skin area was carefully washed with water without altering the existing response, or the integrity of the epidermis. The surrounding untreated skin served as control. In the first step only one animal was used and three test patches were applied successively to this animal. As no serious skin reactions were observed after three minutes, the second patch was applied and removed after one hour. At this stage the observations indicated that with respect to animal welfare the exposure can be allowed to extend to four hours, therefore the third patch was applied and removed after four hours and the responses were graded one hour later. The test was completed using two additional animals, exposed for four hours. The dermal irritation was scored approximately at 1, 24, 48 and 72 hours after patch removal. The dermal irritation was observed after 72 hours therefore the animals were monitored usually on day 7 and 14 after patch removal until the changes had completely subsided. The degree of erythema/eschar formation and oedema formation was recorded as specified by Draize. Any serious lesions or toxic effects other than dermal irritation were also recorded and fully described. As general criteria the body weight of each animal was recorded at the beginning of the study. If clinical findings other than dermal irritations occur they were recorded daily. The mean irritation scores for erythema (redness) and eschar formation at the 24 -h, 48 -h and 72 -h reading points were 2.3, 2.0 and 2.0 for animal 1, 2 and 3, respectively. The mean oedema scores were 2.0, 1.3 and 1.0 for animal 1, 2 and 3, respectively. All skin reactions were fully reversible within 7 days. There were no systemic intolerance reactions. The irritation scores in 2 of 3 animals do not exceed the cut-off values defined in the CLP, therefore the classification as skin irritant is not warranted for this substance.

The supporting study was undertaken in 1999 (Kern, 1999a, according to OECD 404) to investigate the skin irritation potential of the calcium sulfonate read across substance (CAS 70024-69-0) 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.

Furthermore, another key study also was conducted to evaluate the skin irritation potential of the calcium sulfonate read across substance (CAS 70024-69-0; Swan, 1972). In this primary dermal irritation 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 a 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 washing 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%) of 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 a 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, 1981). 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, TG, 1999). 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, Benzenesulfonic acid, di-C10 -14 -alkyl derivs., calcium salts is not classified as a skin irritant in accordance with EU CLP Regulation (EC) No. 1272/2008.

Eye irritation:

In the key study, the irritant effects of the target substance (Benzenesulfonic acid, di-C10-14-alkyl derivs., calcium salts) on eyes of albino rabbits (strain Crl:KBL(NZW)BR) were studied (Gmelin, 2008). The study was conducted according to OECD TG 405 and GLP and is assigned as Klimish 1. 0.1 mL of the pure liquid test substance was placed into the conjunctival sac of one eye of the first animal after having gently pulled the lower lid away from the eyeball. The lids were gently held together for about one second in order to prevent loss of the test compound. The other eye, which remained untreated, served as control. The eye was rinsed approximately 24 hours following instillation. There was no severe irritation observed in the first animal, therefore two further rabbits were treated in the same manner. Eye irritations were scored and recorded approximately at 1, 24, 48 and 72 hours post application. Irritation indices were observed after 72 h, therefore the animals were monitored on day 7, 14 and 21 after application until the changes had completely subsided. The degree of ocular lesions was recorded as specified by Draize. The body weight of each animal was recorded at the beginning of the study. If clinical findings other than eye irritations occur they were recorded daily.

The mean scores at 24 -h, 48 -h, and 72 -h reading time points were 0.0 for corneal opacity and iritis for three animals. Redness conjunctivae scores were 1.3, 1.3 and 1.0 and chemosis conjunctivae scores were 0.3, 1.0 and 0.3 for animal 1, 2, and 3, respectively. All irritation reactions were fully reversible within 7 days. There were no systemic intolerance reactions. According to the classification criteria (CLP/GHS), the substance is not irritating to eyes.

In the supporting study, the eye irritation potential of the calcium sulfonate read across substance, (CAS 70024-69-0), was evaluated in accordance with EPA OPPTS 870.2400 (Kern, 1999b). 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 and were 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 the other supporting 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.

In a supporting 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 the definitive supporting study of the calcium sulfonate read across substance, CAS 70024-69-0, that was conducted following the screening study of Buehler (1990b), 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 (Buehler, 1991b). 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 supporting 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 a supporting 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 benzensulfonic acid, di-C10 -14 -alkyl derivs., calcium salts is not classified as an eye irritating in accordance with EU CLP Regulation (EC) No. 1272/2008.

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

Justification for selection of eye irritation endpoint:
best study available

Justification for classification or non-classification

In vivo studies with the target substance Benzenesulfonic acid, di-C10-14-alkyl derivs., calcium salts and calcium sulfonate read across substances (CAS 70024 -69 -0 and CAS 61789 -86 -4) showed the substances not to be irritating to the skin or the eyes. The skin and eye irritation scores are all under the cut-off values triggering classification and labelling. Therefore, benzensulfonic acid, di-C10 -14 -alkyl dervis., calcium salts does not meet the criteria for classification and labelling for skin or eye irritation in accordance with European regulation (EC) No. 1272/2008.