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Description of key information

Skin sensitisation: negative

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

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Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
The similarity of the substance with the representative samples of sulfated oils is detailed in section 13.2.
The substance was investigated within a screening testing programme using the LLNA method, together with the other representative samples of sulfated oils, and an ambiguous weakly positive result was found for the substances, providing further evidence that their behaviour is similar.
Given the concerns of the applicability of the LLNA method for these substances, a confirmatory test was undertaken of the representative sample with the highest SI in the LLNA studies (CSO3), using the OECD method described in guideline no. 406. Given the above, the result of this further study is considered to be valid for all the sulfated oils considered, including the substance.
Reason / purpose:
read-across source
Reason / purpose:
read-across: supporting information
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Qualifier:
according to
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
A screening testing programme was carried out on seven sulfated oils representative samples using the Local Lymph Node Assay (LLNA: BrdU-ELISA method). Ambiguous results were found, suggesting weakly "positive" outcomes in the absence of QSAR flags for protein-binding (OECD toolbox) implying a false positive response. Thus, a confirmatory assay was undertaken using the GPMT on the substance for which the highest SI value was fouind in the LLNA testing programme (CSO3, SI = 6.27).
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River France
- Age at study initiation: 6 to 7 weeks
- Weight at study initiation: 403-480 g
- Housing: Up to 5 animals/cage noryl cages measuring 74.3 x 54.3 x 25 cm
- Diet (e.g. ad libitum): 8GP17 (Mucedola Srl) diet available ad libitum
- Water (e.g. ad libitum): tap water available ad libitum
- Acclimation period: at least 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2
- Humidity (%): 55 ± 15
- Air changes (per hr): 15 - 20
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 2012-08-27 To: 2012-10-06
Route:
intradermal and epicutaneous
Vehicle:
corn oil
Concentration / amount:
- Concentrations used for induction: intradermal treatment: 0.5% in water; dermal treatment: 5 % in water
- Concentration in Freunds Complete Adjuvant (FCA): 0.5% test substance in a mixture of FCA and water (1:1)
- Concentrations used for challenge: 1 %
Route:
epicutaneous, occlusive
Vehicle:
corn oil
Concentration / amount:
- Concentrations used for induction: intradermal treatment: 0.5% in water; dermal treatment: 5 % in water
- Concentration in Freunds Complete Adjuvant (FCA): 0.5% test substance in a mixture of FCA and water (1:1)
- Concentrations used for challenge: 1 %
No. of animals per dose:
test group: 20 animals
control group: 10 animals
Details on study design:
RANGE FINDING TESTS:
A dose range finding was performed in a preliminary study. 0.1 ml of test substance in water at various concentrations were injected intracutaneously into the skin of the scapula region. 24 hours and 7 days after application skin reaction was assessed.
For dermal applications various concentrations of the substance in water were applied on patches of 2x2 cm to the clipped skin. The patches were removed after 24 hours and dermal reactions were read immediately, 24 and 48 hours after patch removal.


MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2
- Exposure period: 1st exposure: intracutaneous injections (0.1 cm³); 2nd exposure: epicutaneous for 48 hours; evaluation after 24 hours each
- Concentration in Freunds Complete Adjuvants (FCA): test substance in vehicle at 0.5% concentration and in a mixture of FCA and vehicle (1:1)
- Test group: dermal: 5 % test substance in water (2 x 4 cm patch)
- Control group: vehicle (2 x 4 cm patch)
- Frequency of applications: day 1: intradermal treatment, day 8: dermal treatment
- Duration: 3 weeks


B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: on day 22
- Exposure period: dermal application for 24 hours
- Test groups: test substance (2 x 2 cm patch)
- Control group: test substance (2 x 2 cm patch)
- Site: left flanks
- Concentrations: 1 %
- Evaluation: 24 and 48 hours after removal of dressings
Challenge controls:
see above B. Challenge exposure
Positive control substance(s):
yes
Remarks:
alfa-HEXYLCINNAMALDEHYDE
Positive control results:
REFERENCE SUBSTANCE: alfa-HEXYLCINNAMALDEHYDE

CONCENTRATION: INDUCTION (INJECTION) - 20% in corn oil
(TOPICAL) - 50% in corn oil
CHALLENGE - 10% in acetone
CRITICAL DATES: INDUCTION (INJECTION) - 21 August 2012
(TOPICAL) - 28 August 2012
CHALLENGE - 11 September 2012

RESULTS: 70% response in test group and 0% response in control group at challenge.

INTERPRETATION: Incidence at challenge acceptable. Test system regarded as valid.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
1 %
No. with + reactions:
0
Total no. in group:
9
Clinical observations:
no visible change
Remarks on result:
no indication of skin sensitisation
Remarks:
Responses observed 24 h after challange by 24 h topical exposure to test item at 1% concentration and vehicle alone
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
1 %
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no visible change
Remarks on result:
no indication of skin sensitisation
Remarks:
Responses observed 24 h after challange by 24 h topical exposure to test item at 1% concentration and vehicle alone
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
1 %
No. with + reactions:
0
Total no. in group:
9
Clinical observations:
no visible change
Remarks on result:
no indication of skin sensitisation
Remarks:
Responses observed 48 h after challange by 24 h topical exposure to test item at 1% concentration and vehicle alone
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
1 %
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no visible change
Remarks on result:
no indication of skin sensitisation
Remarks:
Responses observed 48 h after challange by 24 h topical exposure to test item at 1% concentration and vehicle alone
Interpretation of results:
GHS criteria not met
Remarks:
not sensitising
Conclusions:
Delayed dermal sensitisation has been investigated using the maximisation test according to OECD/EU test guideline No. 406. The substance did not induce delayed dermal sensitisation in the guinea pig.
Executive summary:

Delayed dermal sensitisation has been investigated using the maximisation test according to OECD/EU test guideline No. 406. The substance did not induce delayed dermal sensitisation in the guinea pig.

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose:
reference to other study
Reason / purpose:
reference to other study
Reason / purpose:
reference to other study
Reason / purpose:
reference to other study
Reason / purpose:
reference to other study
Reason / purpose:
reference to other study
Reason / purpose:
reference to other study
Reason / purpose:
reference to other study
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Qualifier:
according to
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
A screening testing programme was carried out on seven sulfated oils representative samples using the Local Lymph Node Assay (LLNA: BrdU-ELISA method). Ambiguous results were found, suggesting weakly "positive" outcomes in the absence of QSAR flags for protein-binding (OECD toolbox) implying a false positive response. Thus, a confirmatory assay was undertaken using the GPMT on the substance for which the highest SI value was fouind in the LLNA testing programme (CSO3, SI = 6.27).
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River France
- Age at study initiation: 6 to 7 weeks
- Weight at study initiation: 403-480 g
- Housing: Up to 5 animals/cage noryl cages measuring 74.3 x 54.3 x 25 cm
- Diet (e.g. ad libitum): 8GP17 (Mucedola Srl) diet available ad libitum
- Water (e.g. ad libitum): tap water available ad libitum
- Acclimation period: at least 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2
- Humidity (%): 55 ± 15
- Air changes (per hr): 15 - 20
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 2012-08-27 To: 2012-10-06
Route:
intradermal and epicutaneous
Vehicle:
corn oil
Concentration / amount:
- Concentrations used for induction: intradermal treatment: 0.5% in water; dermal treatment: 5 % in water
- Concentration in Freunds Complete Adjuvant (FCA): 0.5% test substance in a mixture of FCA and water (1:1)
- Concentrations used for challenge: 1 %
Route:
epicutaneous, occlusive
Vehicle:
corn oil
Concentration / amount:
- Concentrations used for induction: intradermal treatment: 0.5% in water; dermal treatment: 5 % in water
- Concentration in Freunds Complete Adjuvant (FCA): 0.5% test substance in a mixture of FCA and water (1:1)
- Concentrations used for challenge: 1 %
No. of animals per dose:
test group: 20 animals
control group: 10 animals
Details on study design:
RANGE FINDING TESTS:
A dose range finding was performed in a preliminary study. 0.1 ml of test substance in water at various concentrations were injected intracutaneously into the skin of the scapula region. 24 hours and 7 days after application skin reaction was assessed.
For dermal applications various concentrations of the substance in water were applied on patches of 2x2 cm to the clipped skin. The patches were removed after 24 hours and dermal reactions were read immediately, 24 and 48 hours after patch removal.


MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2
- Exposure period: 1st exposure: intracutaneous injections (0.1 cm³); 2nd exposure: epicutaneous for 48 hours; evaluation after 24 hours each
- Concentration in Freunds Complete Adjuvants (FCA): test substance in vehicle at 0.5% concentration and in a mixture of FCA and vehicle (1:1)
- Test group: dermal: 5 % test substance in water (2 x 4 cm patch)
- Control group: vehicle (2 x 4 cm patch)
- Frequency of applications: day 1: intradermal treatment, day 8: dermal treatment
- Duration: 3 weeks


B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: on day 22
- Exposure period: dermal application for 24 hours
- Test groups: test substance (2 x 2 cm patch)
- Control group: test substance (2 x 2 cm patch)
- Site: left flanks
- Concentrations: 1 %
- Evaluation: 24 and 48 hours after removal of dressings
Challenge controls:
see above B. Challenge exposure
Positive control substance(s):
yes
Remarks:
alfa-HEXYLCINNAMALDEHYDE
Positive control results:
REFERENCE SUBSTANCE: alfa-HEXYLCINNAMALDEHYDE

CONCENTRATION: INDUCTION (INJECTION) - 20% in corn oil
(TOPICAL) - 50% in corn oil
CHALLENGE - 10% in acetone
CRITICAL DATES: INDUCTION (INJECTION) - 21 August 2012
(TOPICAL) - 28 August 2012
CHALLENGE - 11 September 2012

RESULTS: 70% response in test group and 0% response in control group at challenge.

INTERPRETATION: Incidence at challenge acceptable. Test system regarded as valid.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
1 %
No. with + reactions:
0
Total no. in group:
9
Clinical observations:
no visible change
Remarks on result:
no indication of skin sensitisation
Remarks:
Responses observed 24 h after challange by 24 h topical exposure to test item at 1% concentration and vehicle alone
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
1 %
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no visible change
Remarks on result:
no indication of skin sensitisation
Remarks:
Responses observed 24 h after challange by 24 h topical exposure to test item at 1% concentration and vehicle alone
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
1 %
No. with + reactions:
0
Total no. in group:
9
Clinical observations:
no visible change
Remarks on result:
no indication of skin sensitisation
Remarks:
Responses observed 48 h after challange by 24 h topical exposure to test item at 1% concentration and vehicle alone
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
1 %
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no visible change
Remarks on result:
no indication of skin sensitisation
Remarks:
Responses observed 48 h after challange by 24 h topical exposure to test item at 1% concentration and vehicle alone
Interpretation of results:
GHS criteria not met
Remarks:
not sensitising
Conclusions:
Delayed dermal sensitisation has been investigated using the maximisation test according to OECD/EU test guideline No. 406. The substance did not induce delayed dermal sensitisation in the guinea pig.
Executive summary:

Delayed dermal sensitisation has been investigated using the maximisation test according to OECD/EU test guideline No. 406. The substance did not induce delayed dermal sensitisation in the guinea pig.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

The potential for dermal sensitisation of the substance and the other representative substances of sulfated oils has been evaluated using OECD test methods (OECD 442B - Skin sensitisation: Local lymph node assay: BrdU-ELISA). Treatment resulted in an increase in cell proliferation of draining lymph nodes in treated animals when compared to controls, indicating that these substances may elicit a sensitisation response in mice. The ambiguous weakly positive results, unexpected given the lack of any structural alerts for sensitisation, were considered unreliable against a confirmatory assay, using the guinea pig maximisation test, which resulted in negative findings.

It is possible that the specific LLNA method used is not suited to this class of substance (see also Kreiling et al. (2008) Food Chem Toxicol 46: 1896 -1904), QSAR (OECD Toolbox) giving no structural alerts for sensitisation arising from protein binding.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

The available information indicate that the substance does not meet the creiteria for classification as skin sensitising according to Regulation 1272/2008/EC (CLP).