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 (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
17 September 2012 to 25 March 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Fully GLP compliant and in accordance with current test guidelines
Qualifier:
according to
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
GLP compliance:
yes
Type of study:
mouse local lymph node assay (LLNA)
Species:
mouse
Strain:
other: CBA/CaCrl
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Ltd, Margate.
- Age at study initiation: 8 to 11 weeks
- Weight at study initiation: 18 to 21 g
- Housing: The animals were group housed during acclimatisation and individually housed from Day –1 in cages that conformed to the 'Code of Practice for the Housing and Care of Animals Used in Scientific Procedures' (Home Office, London, 1989).
- Diet (e.g. ad libitum): SQC(E) Rat and Mouse Maintenance Diet No 1, ad libitum
- Water (e.g. ad libitum): Mains water, ad libitum
- Acclimation period: 8 to 22 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 24°C
- Humidity (%): 45 to 65%
- Air changes (per hr): The animal rooms were designed to permit 15 to 20 air changes per hour.
- Photoperiod (hrs dark / hrs light): The rooms were illuminated by fluorescent strip-lights for twelve hours daily.

IN-LIFE DATES: From: 18 September 2012 To: 09 October 2012
Vehicle:
acetone/olive oil (4:1 v/v)
Concentration:
Group 1: Vehicle control
Group 2: Low concentration (10% w/v)
Group 3: Intermediate concentration (25% w/v)
Group 4: High concentration (50% w/v)
No. of animals per dose:
4 animals per group
Details on study design:
RANGE FINDING TESTS:
- Compound solubility: Not reported
- Irritation: No irritation noted

MAIN STUDY
The four groups of four female mice were subjected to application of the vehicle or one of the test formulations to the outer aspect of the auditory pinnae, once daily on Days 1, 2 and 3.
On Day 6 the mice were placed in a warming cabinet in order to dilate the peripheral blood vasculature and thus facilitate intravenous dosing. Each mouse was transferred to a cylindrical restrainer. A plastic syringe and fine gauge hypodermic needle were used to administer 0.25 mL phosphate buffered saline incorporating 20 μCi of 3HTdR into a tail vein of each mouse by slow bolus injection. After this treatment the mice were returned to their cages.
Approximately five hours after intravenous injection of the 3HTdR, all mice were killed by exposure to a rising concentration of carbon dioxide. Killing was organised to minimise the interval between death and the recovery of the auricular lymph nodes to no more than fifteen minutes.

TREATMENT PREPARATION AND ADMINISTRATION:
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)
mercaptobenzothiazole (CAS No 149-30-4)
Statistics:
Not applicable
Positive control results:
Positive control not included in this study but checked at least every six months.
Parameter:
SI
Remarks on result:
other: Scintillation fluid with 5% w/v trichloroacetic acid: NA Vehicle control: NA Test article, 10% w/v: 11.7 SI Test article, 25% w/v: 13.3 SI Test article, 50% w/v: 12.7 SI
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: Scintillation fluid with 5% w/v trichloroacetic acid: 65 DPM Vehicle control: 2538 DPM Test article, 10% w/v: 29668 DPM Test article, 25% w/v: 33609 DPM Test article, 50% w/v: 32310 DPM
Interpretation of results:
Category 1 (skin sensitising) based on GHS criteria
Remarks:
Migrated information
Conclusions:
The Local Lymph Node Assay demonstrated that TOFA_TETA_PAA_BADGE_CGE_Adduct has the potential to cause skin sensitisation.
The test article was classified as a Category 1 sensitiser according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS).
Executive summary:

This study was conducted to assess the potential of the test article, TOFA_TETA_PPA_BADGE_BGE_Adduct, to cause skin sensitisation in the mouse.

Following a preliminary screening test using a 50% w/v formulation, the test article was prepared for administration at 10, 25 and 50% w/v in 80% v/v acetone in olive oil. Groups of four female CBA/CaCrl mice were subjected to topical applications of vehicle or of the test formulations to the outer aspect of the auditory pinnae once daily on Days 1, 2 and 3. On Day 6 a 20 µCi dose of tritiated 3H-methyl thymidine was injected intravenously into each mouse. Approximately five hours later the auricular lymph nodes were recovered from each animal. The nodes from mice subjected to the same treatment were pooled and suspensions of the cellular components of the lymph node were prepared in 5% w/v trichloroacetic acid and processed through a scintillation counter.

Test results are expressed in terms of Stimulation Indices, the ratios of the mean scintillation count per group obtained from the test groups relative to the corresponding mean scintillation count from the controls. The threshold level for the Stimulation Index to be considered a positive indicator of the potential to cause skin sensitisation is 3.0.

 

Concentration of test article in applied formulation (% v/v)

 

10%

25%

50%

Stimulation Index

15.2

18.2

17.4

 

The Local Lymph Node Assay demonstrated that TOFA_TETA_PAA_BADGE_BGE_Adduct has the potential to cause skin sensitisation.

The test article was classified as a Category 1 sensitiser according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS).

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

Skin Sensitisation

A LLNA study was conducted to assess the potential of the test article, TOFA_TETA_PAA_BADGE_BGE_Adduct, to cause skin sensitisation in the mouse. Following a preliminary screening test using a 50% w/v formulation, the test article was prepared for administration at 10, 25 and 50% w/v in 80% v/v acetone in olive oil. Groups of four female CBA/CaCrlmice were subjected to topical applications of vehicle or of the test formulations to the outer aspect of the auditory pinnae once daily on Days 1, 2 and 3. On Day 6 a 20 µCi dose of tritiated 3H-methylthymidine was injected intravenously into each mouse. Approximately five hours later the auricular lymph nodes were recovered from each animal. The nodes from mice subjected to the same treatment were pooled and suspensions of the cellular components of the lymph node were prepared in 5% w/v trichloroacetic acid and processed through a scintillation counter.

Test results are expressed in terms of Stimulation Indices, the ratios of the mean scintillation count per group obtained from the test groups relative to the corresponding mean scintillation count from the controls. The threshold level for the Stimulation Index to be considered a positive indicator of the potential to cause skin sensitisation is 3.0.

 

Concentration of test article in applied formulation (% v/v)

 

10%

25%

50%

Stimulation Index

15.2

18.2

17.4

 

The Local Lymph Node Assay demonstrated that TOFA_TETA_PAA_BADGE_BGE_Adduct has the potential to cause skin sensitisation.



Migrated from Short description of key information:
The Local Lymph Node Assay demonstrated that TOFA_TETA_PAA_BADGE_CGE_Adduct has the potential to cause skin sensitisation.

Justification for selection of skin sensitisation endpoint:
Only study available for this endpoint

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

The Local Lymph Node Assay demonstrated that TOFA_TETA_PAA_BADGE_CGE_Adduct has the potential to cause skin sensitisation. Therefore, the substance was classified for Skin Sensitisation Category 1 according to CLP (Regulation EC No 1272/2008) respectively Xi, R43 according to DSD (Directive 67/548/EEC).

Data on respiratory sensitisation are not available and hence the substance is not classified for respiratory sensitisation.