Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2008-02-12 till 2008-03-04
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008
Report Date:
2008

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of study:
mouse local lymph node assay (LLNA)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): FAT 40840/A TE
- Substance type: colouring dye
- Physical state: solid, dark red powder
- Analytical purity: approx. 78.9% org. part (NA-salt), MC: 37.7%, Oligomers: 19.2%
- Lot/batch No.: ROE 358 BOP 01/07
- Expiration date of the lot/batch: June 30, 2012
- Storage condition of test material: At room temperature at about 20°C

In vivo test system

Test animals

Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Netherlands, B.V. Postbus 6174, NL-5960 AD Horst/The Netherlands
- Age at study initiation: 8-9 weeks
- Weight at study initiation: 19.7 - 20.9g
- Housing: single
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: At least 5 days prior to the start of dosing under test conditions after health examination. Only animals without any visible signs of illness were used for the study.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3°C
- Humidity (%): 30-70%
- Photoperiod (hrs dark / hrs light): 6.00 a..m. - 6.00 p.m.


Study design: in vivo (LLNA)

Vehicle:
other: Ethanol:deionised water, 30+70
Concentration:
30:70
No. of animals per dose:
4
Details on study design:
RANGE FINDING TESTS:
- Compound solubility: A solubility experiment was performed according to the recommendations given by OECD 429. The highest test item concentration, which can be technically used was a 20 % solution in ethanol:deionised water (30+70).

To determine the highest non-irritant test concentration, a pre-test was performed in two
animals. Two mice were treated with concentrations of 2.5, 5, 10, and 20 % on one ear
each on three consecutive days. Clinical signs were recorded 24 ± 4 hours after each
application. At the tested concentrations the animals did not show any signs of irritation or
systemic toxicity.

The test item in the main study was assayed at 5, 10, and 20%. The top dose is the
highest technically achievable concentration whilst avoiding systemic toxicity and
excessive local irritation. No severe irritant effects were tolerated choosing the test
concentrations.





MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT

- Name of test method:OECD Guidelines for Testing of Chemicals, Updated Guideline 429: Skin Sensitisation:
Local Lymph Node Assay (adopted 24 April 2002).

- Criteria used to consider a positive response:A test item is regarded as a sensitiser in the LLNA if the following criteria are fulfilled:
- First, that exposure to at least one concentration of the test item resulted in an
incorporation of 3HTdR at least 3-fold or greater than that recorded in control mice, as
indicated by the stimulation index.
- Second, that the data are compatible with a conventional dose response, although
allowance must be made (especially at high topical concentrations) for either local
toxicity or immunological suppression.

Results and discussion

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: concentration: S.I. 5 1.31 10 1.65 20 1.53
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: concentrations: Measurement DPM: control: 3715 5: 4858 10: 6101 20: 5671

Any other information on results incl. tables

Viability/Mortality:

No deaths occurred during the study period.

Clinical Signs:

The animals did not show any clinical signs during the course of the study and no cases of mortality were observed.

Body Weights:

The body weight of the animals, recorded prior to the first application and prior to treatment with 3HTdR, was within the range commonly recorded for animals of this strain and age.

Calculation and results of individual data

 Test Item concentration % (w/v)  Group  Measurement DPM   Calculation         Result
       DPM-BG a)    number of lymph nodes DPM per lymph node b)   S.I. 
---  BG I  21  --- ---  ---  --- 
--- BG II  26  ---  ---  --- --- 
---  3715  3692  8 461.4   
 5 4858  4835  604.3   1.31 
 10 6101  6078  759.7   1.65 
20  5671  5648  705.9   1.53 

BG = Background (1 ml 5% trichloroacetic acid) in duplicate

1 = Control Group

2-4 = Test Group S.I. = Stimulation Index

a) = The mean value was taken from the figures BG I and BG II

b) = Since the lymph nodes of the animals of a dose group were pooled, DPM/node was determined by dividing the measured value by the number of lymph nodes pooled

The EC3 value could not be calculated, since all S.I.´s are below 3.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
The test item FAT 40840/A TE was found to be not a skin sensitiser under the described conditions.
Executive summary:

The animals did not show any clinical signs during the course of the study and no cases of mortality were observed. A test item is regarded as a sensitiser in the LLNA if the exposure to one or more test concentration resulted in 3-fold or greater increase in incorporation of 3HTdR compared with concurrent controls, as indicated by the Stimulation Index (S.I.). The estimated concentration of test item required to produce a S.I. of 3 is referred to as the EC3 value. In this study Stimulation Indices of 1.31, 1.65 and 1.53 were determined with the test item at concentrations of 5, 10, and 20% in ethanol:deionised water (30+70). The EC3 value could not be calculated, since none of the tested concentrations induced an S.I. greater than 3.