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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
7th October 2014 - 23rd October 2014
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
Substance in solvent to make test feasable. The solvent is not thought to impact the results of the test.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report date:
2016

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Version / remarks:
Adopted 22nd July 2010
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of study:
mouse local lymph node assay (LLNA)

Test material

Constituent 1
Chemical structure
Reference substance name:
Reaction products of triethanolamine esters of polyphosphoric acids with alkyl derivatives of pyridine
EC Number:
939-981-3
IUPAC Name:
Reaction products of triethanolamine esters of polyphosphoric acids with alkyl derivatives of pyridine
Constituent 2
Chemical structure
Reference substance name:
Ethanol
EC Number:
200-578-6
EC Name:
Ethanol
Cas Number:
64-17-5
Molecular formula:
C2H6O
IUPAC Name:
ethanol
Test material form:
liquid
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature in the dark
- Stability under storage conditions: Stable
- Stability under test conditions: Stable
- Solubility and stability of the test substance in the solvent/dispersant/vehicle/test medium: Stable

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing:
The test item was used undiluted and freshly prepared as a solution in ethanol/distilled water (7:3).

RADIOLABELLING INFORMATION (if applicable) - phosphate buffered saline (PBS) containing 3H-methyl thymidine
- Radiochemical purity: 3HTdR: 80 μCi/mL
- Specific activity: 2.0 Ci/mmoL

In vivo test system

Test animals

Species:
mouse
Strain:
CBA/Ca
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Females (if applicable) nulliparous and non-pregnant: Yes
- Age at study initiation: Eight to twelve weeks
- Weight at study initiation: 15 to 23g
- Housing: Individually housed in suspended solid floor polypropylene cages furnished with softwood woodflakes
- Diet (e.g. ad libitum): Ad libitum
- Water (e.g. ad libitum): Tapwater ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25°C
- Humidity (%): 30-70%
- Air changes (per hr): 15 per hour
- Photoperiod (hrs dark / hrs light): 12 hour cycles


Study design: in vivo (LLNA)

Vehicle:
other: Ethanol/Distilled water 7:3
Concentration:
Groups of four mice were treated with the undiluted test item or the test item at concentrations of 50% or 25% v/v in ethanol/distilled water 7:3.
No. of animals per dose:
Four
Details on study design:
PRE-SCREEN TESTS:
- Compound solubility:
- Irritation/Systemic toxicity: The preliminary screening test suggested that the test item would not produce systemic toxicity or excessive local skin irritation at the highest suitable concentration.
- Ear thickness measurements: The thickness of each ear was measured using a Mitutoyo 547-300S gauge (MitutoyoCorporation), pre-dose on Day 1, post dose on Day 3 and on Day 6. Any changes in the earthickness were noted. Mean ear thickness changes were calculated between time periods Days 1 and 3 and Days 1 and 6. A mean ear thickness increase of equal to or greater than 25% was considered to indicate excessive irritation and limited biological relevance to the endpoint of sensitization.

MAIN STUDY

ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: Local Lymph Node Assay
- Criteria used to consider a positive response: The test item is regarded as a sensitizer if at least one concentration of the test item results in a threefold or greater increase in 3HTdR incorporation compared to control values

TREATMENT PREPARATION AND ADMINISTRATION:

The mice were treated by daily application of 25 μL of the appropriate concentration of the test item to the dorsal surface of each ear for three consecutive days. The test item formulation was administered using an automatic micropipette and spread over the dorsal surface of the ear using the tip of the pipette. Five days following the first topical application of the test item or vehicle (Day 6) all mice were injected via the tail vein with 250 μL of phosphate buffered saline (PBS) containing 3H-methylthymidine.

Results and discussion

In vivo (LLNA)

Resultsopen allclose all
Key result
Parameter:
SI
Value:
8.56
Test group / Remarks:
25%
Key result
Parameter:
SI
Value:
14.88
Test group / Remarks:
50%
Key result
Parameter:
SI
Value:
11.3
Test group / Remarks:
100%
Cellular proliferation data / Observations:
Sticky, dark brown colored residual test item on the ears was noted in all test animals post dose on Days 1 to 3. Fur loss was noted post dose on Days 1 to 3 and on Days 4 to 6, in animals treated with the undiluted test item or the test item at a concentration of 50% v/v in ethanol/distilled water 7:3.
There were no deaths. No signs of systemic toxicity were noted in the test or control animals during the test.

Applicant's summary and conclusion

Interpretation of results:
Category 1 (skin sensitising) based on GHS criteria
Conclusions:
The results were positive for skin sensitisation in all tested concentrations.
Executive summary:

The test item was considered to be a sensitizer (category 1) under the conditions of the test.