Registration Dossier

Toxicological information

Skin sensitisation

Currently viewing:

Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
01 November 2004-24 November 2004
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study performed to GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2005
Report Date:
2005

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Qualifier:
according to
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
GLP compliance:
yes (incl. certificate)
Type of study:
mouse local lymph node assay (LLNA)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Specific details on test material used for the study:
Sponsors ID: TCPP
Description: pale straw coloured liquid
Batch no: TCPP 09-21-04
Date received: 30 Sept 2004
Storage conditions: approx 4C in dark

In vivo test system

Test animals

Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: B&K Universal Ltd Hull, UK
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 15-23g
- Housing:individually in solid floor polypropylene cages with softwood flakes
- Diet (e.g. ad libitum): Certified Rat and Mouse Diet 5LF2 ad libitum
- Water (e.g. ad libitum): mains tap water ad libitum
- Acclimation period:minimum of 5 days


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


IN-LIFE DATES: From: 1 Nov 2004 To:24 Nov 2004

Study design: in vivo (LLNA)

Vehicle:
acetone/olive oil (4:1 v/v)
Concentration:
100, 50, 25 and 0% v/v
No. of animals per dose:
four
Details on study design:
In a preliminary test, 25 µl of undiluted TCPP was applied topically to the dorsal surface of the ears of one CBA/Ca mouse for three consecutive days and observations were made up to day 6. No clinical signs were noted. In the main test, groups of CBA/Ca mice were treated with 25 µl undiluted TCPP or concentrations of 50% or 25% v/v in acetone: olive oil 4:1 for three days. A further group of mice received the vehicle alone. Five days following the first topical application, all mice were injected via the tail vein with 250 µl of phosphate buffered saline (PBS) containing a total of 20 µCi 3H-methyl thymidine (specific activity 2.0 Ci/mmol). All mice were terminated five hours after injection.
Draining auricular lymph nodes were excised and pooled for each experimental group. A single cell suspension was prepared and washed by repeated centrifugation and resuspension in PBS. Radioactive material was precipitated by resuspending the cells in 3ml of 5% trichloroacetic acid. HTdR incorporation was determined as dpm using liquid scintillation counting following an overnight incubation.
Proliferation resposne of lymph node cells was expressed as the number of radioactive disintegrations per minute per lymph node and as the ratio of HTdR incorporation into lymph node cells of test nodes relative to that recorded for the control nodes.
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)
Statistics:
Not reported

Results and discussion

Positive control results:
10%v/v, SI =1.52;
25% v/v SI = 2.63
50% v/v SI= 5.07

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: Stimulation indices of 1.55, 1.97 and 1.56 were recorded for concentrations of 25, 50 and 100% v/v, respectively.
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: see Remark
Remarks:
concentration DPM DPM/node Stimulation Index Result vehicle 4594.49 574.31 N/A N/A 25% 7131.78 891.47 1.55 negative 50% 9040.95 1130.12 1.97 negative 100% 7179.46 897.43 1.56 negative

Any other information on results incl. tables

There were no mortalities or clinical observations and all bodyweights were comparable to those of the control animals.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Executive summary:

In a preliminary test, 25 µl of undiluted TCPP was applied topically to the dorsal surface of the ears of one CBA/Ca mouse for three consecutive days and observations were made up to day 6. No clinical signs were noted. In the main test, groups of CBA/Ca mice were treated with 25 µl undiluted TCPP or concentrations of 50% or 25% v/v in acetone: olive oil 4:1 for three days. A further group of mice received the vehicle alone. Five days following the first topical application, all mice were injected via the tail vein with 250 µl of phosphate buffered saline (PBS) containing a total of 20 µCi 3H-methyl thymidine (specific activity 2.0 Ci/mmol). All mice were terminated five hours after injection. Draining auricular lymph nodes were excised and pooled for each experimental group. A single cell suspension was prepared and washed by repeated centrifugation and resuspension in PBS. Radioactive material was precipitated by resuspending the cells in 3ml of 5% trichloroacetic acid. HTdR incorporation was determined as dpm using liquid scintillation counting following an overnight incubation. Proliferation resposne of lymph node cells was expressed as the number of radioactive disintegrations per minute per lymph node and as the ratio of HTdR incorporation into lymph node cells of test nodes relative to that recorded for the control nodes.

There were no mortalities or clinical observations and all bodyweights were comparable to those of the control animals. The substance was notfound to be sensitising.