Benzene, di-C10-14-alkyl derivs., sulfonated, sodium salts

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Remarks:

Guinea Pigs Maximisation Test

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From August 04, 2016 to September 23, 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2600 (Skin Sensitisation)

Deviations:

no

Principles of method if other than guideline:

The study was based on the method described by Magnusson and Kligman, "Allergic Contact Dermatitis in the Guinea Pig - Identification of Contact Allergens" (1970).

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

The maximization test was selected since the test substance contains constituents known to cause false positives in the Local Lymph Node Assay.

Test material

Test material information

Constituent 1

Reference substance name:

Benzene, di-C10-14-alkyl derivs., sulfonated, sodium salts

EC Number:

285-586-8

EC Name:

Benzene, di-C10-14-alkyl derivs., sulfonated, sodium salts

Cas Number:

85117-37-9

Molecular formula:

Molecular formula not applicable for this UVCB.

IUPAC Name:

Benzene, di-C10-14-alkyl derivs., sulfonated, sodium salts

Test material form:

solid: particulate/powder

Details on test material:

- Name of test material (as cited in study report): Benzene, di-C1 0-14-alkyl derivs., sulfonated, sodium salts
- ZS name: ALKYLBENZOLNACHLAUF, SULFONIERT, NA-SALZ
- ZS number: 1344
- Batch: SEALS 2011-104-5-8
- Appearance, colour: brown clear liquid

In vivo test system

Test animals

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

female

Details on test animals and environmental conditions:

Source: Charles River Deutschland, Kisslegg, Germany
Age: young adult animals (approx. 4 weeks old) were selected
Acclimatization period: at least 5 days before the start of treatment
Temperature: 18 to 24°C; Relative humidity: 40 to 70%; a 12-hour light/12-hour dark cycle
Diet: complete maintenance diet for guinea pigs (SSNIFF® Spezialdiäten GmbH, Soest, Germany). In addition, hay (TecniLab-BMI BV, Someren, The Netherlands) was provided at least twice a week Water: free access to tap water

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal

Vehicle:

other: Freunds'Complete Adjuvant with water for injection

Concentration / amount:

0.1 mL/site.
a. 1:1 w/w mixture of Freunds' Complete Adjuvant with water for injection.
b. test substance: 1%
c. 1:1 w/w mixture of the test substance, at twice the concentration used in (b) and Freunds' Complete Adjuvant.

Day(s)/duration:

2

Adequacy of induction:

highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically

Induction 2

Route:

epicutaneous, occlusive

Vehicle:

corn oil

Concentration / amount:

0.5 mL of a 2% test substance

Day(s)/duration:

2

Adequacy of induction:

highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically

Challenge

Route:

epicutaneous, occlusive

Vehicle:

corn oil

Concentration / amount:

2% test substance concentration and the vehicle (0.1 mL each)

Day(s)/duration:

1

Adequacy of challenge:

highest non-irritant concentration

No. of animals per dose:

Experimental group: 10 females
Control group: 5 females
(Females were nulliparous and non-pregnant)

Details on study design:

Based on the results of the preliminary irritation study (cfr "Any additional information on materials and methods incl. tables")
NB. Induction - control animals. The control animals were treated as described for the experimental animals except that, instead of the test substance, the vehicle was administered.
The dressing was removed after 24 hours exposure and the skin cleaned of residual test substance and vehicle using water. The treated sites were assessed for challenge reactions 24 and 48 hours after removal of the dressing.
- Observations:
Mortality/viability: twice daily
Toxicity: at least once daily
Body weights: prior to start and at termination of the study
Necropsy: no necropsy for gross macroscopic examination was performed according to study plan
Irritation: skin reactions were graded according to the following numerical scoring systems (0-4). Furthermore, a description of all other (local) effects was recorded for the epidermal treated skin sites

Challenge controls:

Vehicle alone (0.1 mL)

Positive control substance(s):

no

Results and discussion

Positive control results:

-

In vivo (non-LLNA)

Results

Results 1

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

2%

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

no effects

Remarks on result:

no indication of skin sensitisation

Results 2

Key result

Reading:

1st reading

Hours after challenge:

48

Group:

test chemical

Dose level:

2%

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

no effects

Remarks on result:

no indication of skin sensitisation

Any other information on results incl. tables

Interpretation:

The results for the experimental animals at the challenge application(s) were compared with the results for the control animals. Positive skin reactions (grade 1 or more) were considered signs of sensitization, provided that such reactions are not observed or are less persistent in the control group. Other local effects were also taken into account. A sensitization rate (%) was calculated for each concentration as follows: the number of sensitized animals at one concentration as a proportion of the total number of animals of the experimental group. The results were evaluated according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (including all amendments) and the Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008 on classification, labelling and packaging of items and mixtures (including all amendments).

Observations:

Toxicity / mortality: no mortality occurred and no symptoms of systemic toxicity were observed in the animals of the main study. Body weights: body weights and body weight gain of experimental animals remained in the same range as controls over the study period.

No skin irritation were observed in the control group.

Applicant's summary and conclusion

Conclusions:

Under the study conditions, the test substance did not induce skin sensitisation in Guinea pigs.

Executive summary:

A study was conducted to determine the skin sensitisation potential of the test substance according to OECD Guideline 406, EU Method B6 and EPA OPPTS 870.2600 (Guinea pig maximisation test), in compliance with GLP. The study design was based on the preliminary test results. In the main study, 10 female Guinea pigs were intradermally injected with a 1% concentration and epidermally (occlusive) exposed to a 2% concentration. Five control animals were similarly treated, but with vehicle alone (corn oil). Two weeks after the epidermal application all animals were epidermally challenged with a 2% test substance concentration and the vehicle. In the challenge phase, no skin reactions were evident after the challenge exposure in the experimental and control animals. There was no evidence that the test substance had caused skin hypersensitivity in the guinea pig, since no responses were observed in the experimental animals in response to a 2% test substance concentration in the challenge phase. This result indicated a sensitization rate of 0 per cent. No skin irritation were observed in the control group. No mortality occurred and no symptoms of systemic toxicity were observed in the animals of the main study. Body weights and body weight gain of experimental animals remained in the same range as controls over the study period. Under the study conditions, the test substance did not induce skin sensitisation in Guinea pigs (Latour, 2016). Based on the results, the test substance does not have to be classified and has no obligatory labelling requirement for sensitization by skin contact according to the: Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments), and Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).