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Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
24 to 30 June 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Compliant with OECD 429 and GLP guideline

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report date:
2009

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
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
Details on test material:
Name: Golden Yellow Continuous

In vivo test system

Test animals

Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Species and strain: CBA/J@Rj mice
- Source: ELEVAGE JANVIER, Route des Chènes Secs B.P. 4105, 53940 LE GENEST-ST-ISLE, France
- Age at study initiation: 10-11 weeks
- Weight at study initiation: 21.3–22.8 grams
- Housing: single
- Diet: ssniff SM R/M-Z+H ad libitum
- Water: tap water ad libitum
- Acclimation period: 27 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3°C
- Humidity (%): 30 - 70%
- Air changes (per hr): 15 - 20
- Photoperiod (hrs dark / hrs light): 12/12


IN-LIFE DATES: From: 24 to 30 June 2009

Study design: in vivo (LLNA)

Vehicle:
dimethyl sulphoxide
Concentration:
2.5%, 5%, 10%
No. of animals per dose:
4
Details on study design:
RANGE FINDING TESTS:
- Compound solubility: max 10% in DMSO
- Irritation: no
- Lymph node proliferation response: NA


MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: Lymph node proliferation assay with incorporated tritiated thymidine
- Criteria used to consider a positive response:
-- exposure to at least one concentration resulted in an incorporation of 3HTdR at least 3-fold or greater than recorded in control mice, as indicated by the stimulation index.
AND
-- 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.


TREATMENT PREPARATION AND ADMINISTRATION:
Each mouse was topically dosed with 25 µl of the appropriate formulation (1%, 2.5%, 5%) of the test item using a pipette, on the dorsal surface of each ear.
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)

Results and discussion

Positive control results:
DPM: 8422.5
DPN: 1052.8
SI: 6.6

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: 2.5%: 2.6 5. 0%: 1.7 10%: 1.6
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: 2.5%: 3232.5 5. 0%: 2190.5 10%: 2057.5

Any other information on results incl. tables

Test Group Name

Measured DPM/group

DPM

No. of LNN

DPN

Stimulation Index Values

Background 5 (w/v) % TCA

35.5

-

-

-

-

Negative control DMSO

1303.0

1267.5

8

158.4

1.0

Golden Yellow Continuous
10 (w/v) %

2093.0

2057.5

8

257.2

1.6

Golden Yellow Continuous
5 (w/v) %

2226.0

2190.5

8

273.8

1.7

Golden Yellow Continuous
2.5 (w/v) %

3268.0

3232.5

8

404.1

2.6

Positive control
25 (w/v) % HCA

8458.0

8422.5

8

1052.8

6.6

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
In conclusion, under the conditions of the present assay, Golden Yellow Continuous, tested in a suitable vehicle, proved to have no sensitizing potential (non-sensitizer) in the Local Lymph Node Assay.
Executive summary:

The aim of this study was to determine the skin sensitisation potential of Golden Yellow Continuous following dermal exposure. 

Based on results of the Preliminary Compatibility Test and on recommendations of the OECD Test Guideline 429, the test item was dissolved in Dimethyl-sulphoxide (DMSO). The maximum attainable concentration based on solubility of the test item in DMSO was 10%.

A Preliminary Irritation/Toxicity Test was performed with the test item at concentrations of 10% and 5% in the selected vehicle. According to this test, the applicability and biocompatibility of the test item to the animals’ ears (2 female CBA/J@Rj mice/dose) at the maximum concentration of 10% was acceptable.

 

In the main assay twenty female CBA/J@Rj mice were allocated to five groups of four animals each:

-      three groups received the appropriate formulation of Golden Yellow Continuous at concentrations of 10%, 5% and 2.5%,

-      the negative control group received the vehicle (DMSO),

-      the positive control group received 25% alpha-Hexylcinnamaldehyde (HCA) in DMSO.

 

The test item solutions were applied to the dorsal surface of both ears of the mice (25 µL/ear) for three consecutive days (Days 1, 2, and 3). There was no treatment on Days 4, 5, and 6. On Day 6, the cell proliferation in the local lymph nodes was measured by incorporation of tritiated methyl thymidine (3HTdR) and the values obtained were used to calculate stimulation indices (SI).

 

No mortality or systemic clinical signs were observed during the study. No treatment related effects were observed on animal body weights in any of the treated groups. No cutaneous reactions were observed at the site of the treatment in any of the treated groups.

No significant lymphoproliferative response (SI ³ 3) of Golden Yellow Continuous was observed at the concentrations applied compared to the negative control. The stimulation index values of the test item were 1.6, 1.7, and 2.6 at treatment concentrations of 10%, 5%, and 2.5%, respectively. No conventional dose-response to treatment was observed.

A significant lymphoproliferative response (stimulation index value of 6.6) was noted for the positive control chemical HCA. This result confirmed the validity of the assay.

 

In conclusion, under the conditions of the present assay Golden Yellow Continuous, tested in a suitable vehicle, proved to have no sensitization potential (non-sensitizer) in the Local Lymph Node Assay.