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EC number: 203-783-9 | CAS number: 110-61-2
Skin sensitisation
Administrative data
- Endpoint:
- skin sensitisation: in vivo (LLNA)
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2016
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 016
- Report date:
- 2016
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
- GLP compliance:
- yes (incl. QA statement)
- Type of study:
- mouse local lymph node assay (LLNA)
Test material
- Reference substance name:
- Succinonitrile
- EC Number:
- 203-783-9
- EC Name:
- Succinonitrile
- Cas Number:
- 110-61-2
- Molecular formula:
- C4H4N2
- IUPAC Name:
- butanedinitrile
- Details on test material:
- - Analytical purity: 97%
- Source: Aldrich Chemical Company, Milwaukee, WI, USA
Constituent 1
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material:DSM Engineering Plastics
- Expiration date of the lot/batch:SNO620150908
- Purity test date:100%
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: In refrigerator (2—8°C)
- Stability under test conditions: temp < 150 C
In vivo test system
Test animals
- Species:
- mouse
- Strain:
- CBA:J
- Sex:
- female
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source: Source: Janvier, Le Genest-Saint—lsle, France
- Females (if applicable) nulliparous and non-pregnant: [yes/]
- Age at study initiation:approx. 10 weeks old
- Weight at study initiation: Body weight
variation was within +/- 20% of the sex mean.
- Housing: Animals were group housed in labeled Makrolon cages (Mlll type; height 18 cm) containing sterilised sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Sbhne GmbH + CO. KG,Rosenberg, Germany). Paper (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom) and shelters (disposable paper corner home, MCORN 404, Datesand Ltd, USA) were supplied as cage-enrichment. The acclimatization period was at least 5 days before the start of treatment under laboratory conditions. On Day 6, the animals were group housed in Makrolon Mll type cages with a sheet of paper instead of sawdust and cage enrichment.
- Diet : Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiaten GmbH, Soest, Germany).
- Water : Free access to tap water.
- Acclimation period:
- Indication of any skin lesions: no
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 24°C
- Humidity (%): 40 to 70%,
- Photoperiod (hrs dark / hrs light): 12-hour light/12-hour dark cycle.
Study design: in vivo (LLNA)
- Vehicle:
- dimethylformamide
- Concentration:
- No irritation was observed in any of the pre-screen animals and variations in ear thickness during the
observation period were less than 25% from Day 1 pre-dose values.
Based on these results, the highest test item concentration selected for the main study was a 10%
5% and 10% was considered not
to have a toxicologically significant effect on the activity of the nodes. - No. of animals per dose:
- 5
- Details on study design:
- PRE-SCREEN TESTS:
- Compound solubility:
- Irritation:
- Systemic toxicity:5% and 10% was considered not to have a toxicologically significant effect on the activity of the nodes.
- Ear thickness measurements:
- Erythema scores:
MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method:
- Criteria used to consider a positive response:
TREATMENT PREPARATION AND ADMINISTRATION: - Positive control substance(s):
- hexyl cinnamic aldehyde (CAS No 101-86-0)
- other: The results of a reliability test with three concentrations of Hexylcinnamaldehyde (CAS No. 101-86-0) in Acetone/Olive oil (4:1 v/v), performed not more than 6 months previously and using the same materials, animal supplier, animal strain and essential pr
Results and discussion
- Positive control results:
- Group1
% Alpha- Hexylcinnamaldehyde, technical grade 0% (Acetone:Olive oil (4:1 v/v))
SI value: 1.0 ± 0.2
Group2
% Alpha- Hexylcinnamaldehyde, technical grade 5%
SI value: 1.0 ± 0.2
Group3
% Alpha- Hexylcinnamaldehyde, technical grade 10%
SI value: 1.5 ± 0.2
Group4
% Alpha- Hexylcinnamaldehyde, technical grade 25%
SI value: 4.4 ± 0.2
In vivo (LLNA)
Resultsopen allclose all
- Key result
- Parameter:
- SI
- Value:
- 1.1
- Variability:
- +/-0.2
- Test group / Remarks:
- Group 4
Animal 16-20
Test item concentration 10 w/w%
- Key result
- Parameter:
- SI
- Value:
- 1.1
- Variability:
- +/- 0.2
- Test group / Remarks:
- Group 3
Animal 11-15
Test item concentration 5 w/w%
- Key result
- Parameter:
- SI
- Value:
- 0.8
- Variability:
- +/- 0.2
- Test group / Remarks:
- Group 2
Animal 06-10
Test item concentration 2 w/w%
- Key result
- Parameter:
- SI
- Value:
- 1
- Variability:
- +/- 0.2
- Test group / Remarks:
- Group 1 Reference group (Vehicle N,N-Dimethyl formamide )
animals 01 - 05
Test item concentration 0%
- Key result
- Parameter:
- SI
- Value:
- 0.8
- Variability:
- +/_ 0. 2
Any other information on results incl. tables
8.1. Pre-screen Test
Both animals treated at 25% were found dead on Day 2.
No irritation was observed in any of the pre-screen animals and variations in ear thickness during the
observation period were less than 25% from Day 1 pre-dose values.
Based on these results, the highest test item concentration selected for the main study was a 10%
concentration.
8.2. Main Study
8.2.1. Skin Reactions I Irritation
The slight irritation of the ears as shown by some animals treated at 5% and 10% was considered not
to have a toxicologically significant effect on the activity of the nodes.
8.2.2. Systemic Toxicity
No mortality occurred and no clinical signs 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.
8.2.3. Macroscopic Examination of the Auricular Lymph Nodes and Surrounding Area
All auricular lymph nodes of the animals of the experimental and control groups were considered
normal in size. No macroscopic abnormalities of the surrounding area were noted for any of the
animals.
8.2.4. Radioactivity Measurements and SI Values
Mean DPM/animal values for the experimental groups treated with test item concentrations 2, 5 and
10% were 730, 952 and 974 DPM, respectively. The mean DPM/animal value for the vehicle control
group was 866 DPM. The SI values calculated for the test item concentrations 2, 5 and 10% were 0.8,
1.1 and 1.1, respectively.
Applicant's summary and conclusion
- Interpretation of results:
- GHS criteria not met
- Conclusions:
- Since there was no indication that Succinonitrile could elicit a SI 2 3 when tested up to 10%, it was
established that the EC3 value (the estimated test item concentration that will give a SI =3) (if any)
exceeds 10%.
The six-month reliability check with Alpha-hexylcinnamaldehyde indicates that the Local Lymph Node
Assay as performed at WIL Research Europe is an appropriate model for testing for contact
hypersensitivity (see APPENDIX 2).
Based on these results, Succinonitrile would not be regarded as a skin sensitizer according to the
recommendations made in the test guidelines. The test item 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 (2011) (including all
amendments) and the Regulation (EC) No 1272/2008 on classification, labelling and packaging of
items and mixtures (including all amendments). - Executive summary:
Assessment of skin sensitization to Succinonitrile in the Mouse (Local Lymph Node Assay).
The study was carried out based on the guidelines described in:
OECD, Section 4, Health Effects, No.429 (2010),
EC, No 440/2008; B42: "Skin Sensitization: Local Lymph Node Assay"
EPA, OPPTS 870.2600 (2003) ”Skin Sensitization”.
Test item concentrations selected for the main study were based on the results of a pre-screen test.
In the main study, three experimental groups of five female CBA/J mice were treated with test item
concentrations of 2, 5 or 10% w/w on three consecutive days, by open application on the ears. Five
vehicle control animals were similarly treated, but with the vehicle alone (N,N-Dimethyl formamide).
Three days after the last exposure, all animals were injected with 3H-methyl thymidine and after five
hours the draining (auricular) lymph nodes were excised and pooled for each animal.
After precipitating the DNA of the lymph node cells, radioactivity measurements were performed. The
activity was expressed as the number of disintegrations per minute (DPM) and a stimulation index (SI)
was subsequently calculated for each group.
The slight irritation of the ears as shown by some animals treated at 5% and 10% was considered not
to have a toxicologically significant effect on the activity of the nodes.
All auricular lymph nodes of the animals of the experimental and control groups were considered
normal in size. No macroscopic abnormalities of the surrounding area were noted for any of the
animals.
Mean DPM/animal values for the experimental groups treated with test item concentrations 2, 5 and
10% were 730, 952 and 974 DPM, respectively. The mean DPM/animal value for the vehicle control
group was 866 DPM. The SI values calculated for the test item concentrations 2, 5 and 10% were 0.8,
1.1 and 1.1, respectively.
Since there was no indication that Succinonitrile could elicit a SI 2 3 when tested up to 10%, it was
established that the EC3 value (the estimated test item concentration that will give a Si =3) (if any)
exceeds 10%.
The six-month reliability check with Alpha-hexylcinnamaldehyde indicates that the Local Lymph Node
Assay as performed at WlL Research Europe is an appropriate model for testing for contact
hypersensitivity.
Based on these results, Succinonitrile would not be regarded as a skin sensitizer according to the
recommendations made in the test guidelines. The test item 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 (2011) (including all
amendments) and the Regulation (EC) No 1272/2008 on classification, labelling and packaging of
items and mixtures (including all amendments).
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