Registration Dossier

Administrative data

Description of key information

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records
Reference
Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2012 - 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
Qualifier:
equivalent or similar to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. QA statement)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
It is acknowledged by the registrant that the LLNA has advantages over the GPMT in terms of animal welfare, time need, objectivity of read out and quantitation of results, however, unspecific findings of substance known to be non-sensitizers are published in peer-reviewed scentific literature and cannot be excluded in the LLNA, especially for unsaturated organic substances (see reference literature). For this group of chemicals, the GPMT may give results that more adequately reflect the relevance in humans. (reference literature: Comparison of the skin sensitizing potential of unsaturated compounds asassessed by the murine local lymphnode assay(LLNA)and the guinea pig maximization test(GPMT), Food and Chemical Toxicology 46 (2008) 1896–1904).
Due to the fact that the registered substance contains unsaturated organic components, it was considered justified to conduct a GPMT.
Specific details on test material used for the study:
- Name of test material (as cited in study report): DE07_2012_001_PLSW
- Substance type: Alkylsulfate/alkylethersulfate
- Chemical name: Alcohols, C16-18 (even numbered, C18-unsatd.), ethoxylated, and alcohols C20-22 (even numbered), sulfates, ammonium salts
- CAS: n/a
- Physical state: ivory paste-like solid at 20 °C
- Batch No.: PU20740016, PU20740019
- Purity: 100 % (UVCB)
- Storage condition of test material: Room temperature, protected from light
- Stability: stable under test conditions
Species:
guinea pig
Strain:
other: Crl: HA - Guinea Pigs
Sex:
female
Details on test animals and environmental conditions:
Species/strain: healthy Crl: HA - Guinea Pigs (full barrier)
Source: Charles River, 97633 Sulzfeld, Germany
Sex: female (non-pregnant, nulliparous)
Age at the
beginning of the study: 4 weeks old
Body weight at the
beginning of the study: 324 - 369 g

Housing and Feeding Conditions

- Semi barrier in an air-conditioned room
- Temperature: 22 +/- 3 °C
- Relative humidity: 55 +/- 10 %
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: at least 10 x / hour
- Free access to autoclaved hay and to Altromin 3122 maintenance diet for guinea pigs, rich in crude fibre
- Free access to tap water (drinking water, municipal residue control, microbiological controls at regular intervals)
- The animals were kept in groups in Terluran - cages on Altromin saw fibre bedding
- Certificates of food, water and bedding are filed
- Adequate acclimatisation period (at least five days) under laboratory conditions
Route:
intradermal and epicutaneous
Vehicle:
physiological saline
Concentration / amount:
1% for the intradermal induction
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Route:
epicutaneous, occlusive
Vehicle:
other: vasiline
Concentration / amount:
6.25% for the dermal induction
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
other: vasiline
Concentration / amount:
a concentration of 3% for the challenge application
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
Number of animals in the test group: 10
Number of animals in the negative-control group: 5
Number of animals in the positive control group: 5
Number of animals in the dose range finding study: 8
Details on study design:
Based on the results of this preliminary test the following concentrations were chosen for the main test:
a concentration of 1% for the intradermal induction
a concentration of 6.25% for the dermal induction
a concentration of 3% for the challenge application
Challenge controls:
For the intradermal injection (induction - first stage), 0.2 g of the positive control were suspended in cottonseed oil to gain a final volume of 10 mL of a 2% solution (w/v).
For the topical application (induction – second stage), 2.5 g of the positive control were suspended in vaseline to gain a final volume of 10 g of a 25% suspension (w/w).
For the topical application (challenge), 3 g of the positive control were suspended in vaseline to gain a final volume of 20 g of a 15% suspension (w/w).
Positive control substance(s):
yes
Remarks:
2-Mercaptobenzothiazole
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
3 %
No. with + reactions:
4
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 3 %. No with. + reactions: 4.0. Total no. in groups: 10.0.
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
3 %
No. with + reactions:
1
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 3 %. No with. + reactions: 1.0. Total no. in groups: 10.0.
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
3 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 3 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
3 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 3 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
3 %
No. with + reactions:
5
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: positive control. Dose level: 3 %. No with. + reactions: 5.0. Total no. in groups: 5.0.
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
3 %
No. with + reactions:
5
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: positive control. Dose level: 3 %. No with. + reactions: 5.0. Total no. in groups: 5.0.

1. Induction: First Stage, Intradermal Injection

Test Group: Day 0

Injection 1: a 1:1 mixture (v/v) FCA/physiological saline 0.9% NaCl

Injection 2: a 1% concentration of the test item in physiological saline 0.9% NaCl

Injection 3: a 1% concentration of the test item formulated in a 1:1 mixture (v/v) FCA/physiological saline 0.9% NaCl

Negative Control Group: Day 0

Injection 1: a 1:1 mixture (v/v) FCA/physiological saline 0.9% NaCl

Injection 2: 100% physiological saline 0.9% NaCl

Injection 3: a 50% (v/v) formulation of physiological saline 0.9% NaCl in a 1:1 (v/v)

mixture FCA/physiological saline 0.9% NaCl

Injections 1 and 2 were given close to each other and nearest to the head, while

injection 3 was given toward the caudal part of the test area.

Positive Control Group: Day 0

Injection 1: a 1:1 mixture (v/v) FCA/physiological saline 0.9% NaCl

Injection 2: 2% mercaptobenzothiazole in cottonseed oil

Injection 3: a 50% w/v formulation of mercaptobenzothiazole in a 1:1 (v/v)

mixture FCA/physiological saline 0.9% NaCl

2. Induction: Second Stage, Topical Application

Test Group and Control Groups: Day 6

Approximately twenty-four hours before the topical application the test area was

close-clipped.

Test Group: Day 7

The test item was suspended in vaseline at a concentration of 6.25%. A patch was fully loaded with 0.5 g of the prepared test item. Then it was applied to the test area and held in contact with the help of an occlusive dressing for 48 hours.

Negative Control Group: Day 7

A patch was fully loaded with 0.5 g of vaseline. Then it was applied to the test area and held in contact with the help of an occlusive dressing for 48 hours.

Positive Control Group: Day 7

A patch was fully loaded with 2.0 mL of 2-Mercaptobenzothiazol at a concentration of 25% (vehicle: vaseline), applied to the test area and held in contact by an occlusive dressing for 48 hours.

3. Challenge: Topical Application

The flanks of treated and control animals were cleared of hair by close-clipping.

Test Group and Negative Control Group: Day 20

The test item was suspended with vaseline at a concentration of 3%. A patch loaded with 0.5 g of the prepared test item was applied to the left flank of the animals and a patch loaded with 0.5 g of vaseline to the right flank (intraspecific control). The patches

were held in contact with the help of an occlusive dressing for 24 hours.

Positive Control Group: Day 20

A patch loaded with 2.0 mL of 2-Mercaptobenzothiazol at a concentration of 15% was applied to the left flank of the animals and, a patch loaded with 0.5 g of vaseline to the right flank (intraspecific control), respectively. The patches were held in contact by an occlusive dressing for 24 hours. The application area was not rinsed.

Approximately 24 hours after removing the patch, the challenge area was cleared of hair by the use of a depilatory cream. Approximately 24 and 48 hours after removing the patch the skin reaction was observed and recorded. Additionally all animals were observed for signs of toxicity at least once daily during the test period.

Interpretation of results:
Category 1B (indication of skin sensitising potential) based on GHS criteria
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of the present study it can be stated that the test item caused reactions identified as sensitisation at the tested concentration.
According to the EC criteria for classification and labelling requirements for dangerous substances and preparations (Guidelines in Commission Directive 2001/59/EC) labelling is necessary, as the percentage of sensitised animals was greater than 30%.
According to Commission Regulation (EU) No 286/2011 as well as GHS (Globally Harmonized Classification System) the test item has obligatory labelling requirement for skin sensitisation and is classified into Category 1B.
Executive summary:

Introduction:

The test for sensitisation is performed on the guinea pig. It has been the animal of choice for predictive sensitisation tests for surfactants, e.g. alkylsulfates, alkylethersulfates. The assessment of the test item to produce skin sensitisation is achieved through multiple exposure under standardised conditions.

Method:

For the intradermal injection (induction - first stage), 0.1 g of the test item were dissolved in physiological saline 0.9% NaCl to gain a final volume of 10 mL of a 1% solution (w/v). For the topical application (induction – second stage), 0.625 g of the test item were suspended in vaseline to gain a final volume of 10 g of a 6.25% suspension (w/w). For the topical application (challenge), 0.3 g of the test item were suspended in vaseline to gain a final volume of 10 g of a 3% suspension (w/w).

2-Mercaptobenzothiazole was used as positive control.

Conclusion:

Under the conditions of the present study it can be stated that the test item caused reactions identified as sensitisation at the tested concentration.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (sensitising)
Additional information:

Within a Guinea Pig Maximisation Test according to OECD Guideline 406, 4/10 animals at the 24 h reading and 1/10 animals upon the 48 h reading showed skin reactions after challenge with 3% of the test item. Induction was performed with 1% test substance. Therefore this study represents a borderline result for classification as Skin sensitizing Category 1B (≥30%). It should be mentioned that the study size of 10 animals is low when assuming this borderline result and the fact that the test item showed mild irritating properties within the acute dermal toxicity study as skin irritation is a confounding factor when scoring skin reactions within a skin sensitization study.


Migrated from Short description of key information:
Under the conditions of the present study (GMPT test) it can be stated that the test item caused reactions identified as sensitisation at the tested concentration.

Justification for selection of skin sensitisation endpoint:
Under the conditions of the present study it can be stated that the test item caused reactions identified as sensitisation at the tested concentration.
According to the EC criteria for classification and labelling requirements for dangerous substances and preparations (Guidelines in Commission Directive 2001/59/EC) labelling is necessary, as the percentage of sensitised animals was greater than 30%. According to Commission Regulation (EU) No 286/2011 as well as GHS (Globally Harmonized Classification System) the test item has obligatory labelling requirement for skin sensitisation and is classified into Category 1B.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Under the conditions of the present study it can be stated that the test item caused reactions identified as sensitisation at the tested concentration. According to the EC criteria for classification and labelling requirements for dangerous substances and preparations (Guidelines in Commission Directive 2001/59/EC) labelling is necessary, as the percentage of sensitised animals was greater than 30%. According to Commission Regulation (EU) No 286/2011 as well as GHS (Globally Harmonized Classification System) the test item has obligatory labelling requirement for skin sensitisation and is classified into Category 1B.