Salts of 9-(2-carboxyphenyl)-3,6-bis(diethylamino)xanthenium and 4,4'-bis[(3-methyl-5-oxo-1-phenyl-4,5-dihydro-1H-pyrazol-4-yl)diazo] biphenyl- 2,2'-disulfonic acid and 5-amino-4-hydroxy-6-[(4-nitrophenyl)diazo]-3-(phenyldiazo)-3,4-dihydronaphthalene-2,7-disulfonic acid (5:3:1)

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2008-04-09 to 2008-09-04

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

Deviations:

no

Test guideline 2

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

Test material information

Constituent 1

Reference substance name:

Reaction product of Acid Yellow 42, Acid Black 2 and Basic Violet 1

IUPAC Name:

Reaction product of Acid Yellow 42, Acid Black 2 and Basic Violet 1

Constituent 2

Reference substance name:

Acid Yellow 42: 6375-55-9

IUPAC Name:

Acid Yellow 42: 6375-55-9

Constituent 3

Reference substance name:

Acid Black 2: reaction product of 8005-03-6 and 68510-98-5

IUPAC Name:

Acid Black 2: reaction product of 8005-03-6 and 68510-98-5

Constituent 4

Reference substance name:

Basic Violet 1: 8004-87-3

IUPAC Name:

Basic Violet 1: 8004-87-3

Details on test material:

- Name of test material: Reaction product of Acid Yellow 42, Acid Black 2 and Basic Violet 1
- Lot/batch No.: 2008041541
- Expiration date of the lot/batch: 2010-01-31
- Stability under test conditions: see expiry date
- Storage condition of test material: store cool and dry
- Analytical purity: 100 % (96.8 %; UVCB substance)

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Harlan Hungarian Supplier:
WOBE Kereskedelmi Kft.
H-1164, Budapest, Garmada u. 10.
- Age at study initiation: Young adult, 10-11 weeks old, age-matched within one week
- Weight at study initiation: The weight variation in animals involved in the study did not exceed  20 % of the mean weight.
- Housing: Individual caging
- Diet (e.g. ad libitum): Animals received ssniff® SM R/M-Z+H "Autoclavable complete feed for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany ad libitum
- Water (e.g. ad libitum): tap water from municipal supply, as for human consumption from 500 ml bottle ad libitum
- Acclimation period: 28 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): 8-12 air exchange/hour
- Photoperiod (hrs dark / hrs light): 12 hours light/day

Study design: in vivo (LLNA)

Vehicle:

dimethylformamide

Concentration:

An irritation/toxicity screen was performed in groups of 2 CBA/Ca mice with test item concentrations of 25 % and 10 % in DMF. This preliminary study was conducted in a similar experimental manner to the exposure phase of the main study. The test item concentrations in the main study ranged from 25-5 %.

No. of animals per dose:

4

Details on study design:

RANGE FINDING TESTS:
- Compound solubility: 25 %
- Irritation:
- Lymph node proliferation response:


MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method:
- Criteria used to consider a positive response:


TREATMENT PREPARATION AND ADMINISTRATION:

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Results and discussion

Positive control results:

The positive control substance a-Hexylcinnamaldehyde (HCA) was examined at a concentration of 25 % in the relevant vehicle.

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: The stimulation index values were 14.9, 15.7 and 9.4 at concentrations of 25 %, 10 % and 5 %, respectively. The stimulation index values were compatible with the conventional biological dose-response.

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: F 46 Black (25 % in DMF): 14709.9 (per group) F 46 Black (10 % in DMF): 15498.8 (per group) F 46 Black (5 % in DMF): 9296.2 (per group)

Applicant's summary and conclusion

Interpretation of results:

sensitising

Remarks:

Migrated information

Conclusions:

Under the conditions of the present assay F 46 Black, dissolved in an appropriate solvent up to the maximum feasible concentration (25 %), revealed sensitisation potential (sensitiser) in the local lymph node assay.

Executive summary:

Based on a local lymph node assay according to EU method B.42 and OECD guideline 429 the test item F 46 Black revealed sensitisation properties.