Registration Dossier

Toxicological information

Skin sensitisation

Currently viewing:

Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
secondary literature
Justification for type of information:
Data is from secondary source.

Data source

Reference
Reference Type:
secondary source
Title:
opinion of the scientific committee on cosmetic products and non-food products intended for consumers, CAS NO: 25956-17-6 (curry red)
Author:
European Commission (EC)
Year:
2004
Bibliographic source:
Evaluation and opinion on Curry Red, 2004

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Principles of method if other than guideline:
The skin sensitization study of Disodium 6-hydroxy-5-[(2-methoxy-4-sulphonato-m-tolyl)azo]naphthalene-2-sulphonate was performed on mice by using Mice local lymph node assay (LLNA)
GLP compliance:
not specified
Type of study:
mouse local lymph node assay (LLNA)

Test material

Constituent 1
Chemical structure
Reference substance name:
Disodium 6-hydroxy-5-[(2-methoxy-4-sulphonato-m-tolyl)azo]naphthalene-2-sulphonate
EC Number:
247-368-0
EC Name:
Disodium 6-hydroxy-5-[(2-methoxy-4-sulphonato-m-tolyl)azo]naphthalene-2-sulphonate
Cas Number:
25956-17-6
Molecular formula:
C18H16N2O8S2.2Na
IUPAC Name:
disodium 6-hydroxy-5-[(2-methoxy-3-methyl-4-sulfonatophenyl)diazenyl]naphthalene-2-sulfonate
Details on test material:
Details on test material
- Name of test material (as cited in study report): Curry Red
- Molecular formula: C18H16N2O8S2.2Na
- Molecular weight:496.426 g/mole
- Substance type: organic
- Physical state: Dark red powder
- Purity: No data available
- Impurities (identity and concentrations): No data available
Specific details on test material used for the study:
Details on test material
- Name of test material (as cited in study report): Curry Red
- Molecular formula: C18H16N2O8S2.2Na
- Molecular weight:496.426 g/mole
- Substance type: organic
- Physical state: Dark red powder
- Purity: No data available
- Impurities (identity and concentrations): No data available

In vivo test system

Test animals

Species:
mouse
Strain:
CBA:J
Sex:
female

Study design: in vivo (LLNA)

Vehicle:
other: aqua/acetone (1:1) and olive oil (4:1)
Concentration:
0.5, 1, 2, 4 % aqua/acetone (1:1) mixed with olive oil (4:1)
No. of animals per dose:
Total:15
Test group :5 female
control group :5 female
positive control group :5 female
Details on study design:
Details on study design

PRE-SCREEN TESTS:
- Compound solubility: No data available
- Irritation: No data available
- Systemic toxicity: No data available
- Ear thickness measurements: No data available
- Erythema scores: No data available

MAIN STUDY

ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: Local Lymph Node Assay (LLNA)
- Criteria used to consider a positive response: measuring the cell proliferation in the draining lymph nodes after topical application onto the ears.

TREATMENT PREPARATION AND ADMINISTRATION: 25 μl of 0 (vehicle only), 0.5, 1, 2 and 4 %of Curry Red in either DMSO or a mixture of aqua/acetone (1:1) with olive oil (4:1) were applied on three consecutive days to the surface of the ear of 5 female CBA/J mice per group.
After application, the ears were dried for about 5 minutes by means of a hair dryer.
A positive control (p-phenylenediamine at 1 % in DMSO) was investigated in parallel under identical tests conditions.
Positive control substance(s):
other: p-phenylenediamine at 1 % in DMSO
Statistics:
The mean dpm per treated group and the stimulation index (test item compared to the concurrent vehicle control)

Results and discussion

Positive control results:
an increase in the stimulation index by a Factor of 7.8.

In vivo (LLNA)

Results
Parameter:
SI
Value:
0.9
Test group / Remarks:
5
Remarks on result:
other: not sensitizing
Cellular proliferation data / Observations:
CLINICAL OBSERVATIONS: Animals were observed daily before and at least once after dosing.
BODY WEIGHTS: Body weight was determined on day 1 and day 5.
mean stimulation index For
a)test substance as 0.5, 1, 2, 4 % (w/v) in DMSO mean stimulation indices of 0.6, 0.7, 0.9 and 0.9 were obtained
b) test substance as 0.5, 1, 2, 4 % in aqua/acetone (1:1) mixed with olive oil (4:1) mean stimulation indices of1.2, 1.0, 0.9 and 0.9 were obtained
c) The positive control (PPD, 1 % in DMSO) caused an increase in the stimulation index by a factor of 7.8.

Applicant's summary and conclusion

Interpretation of results:
other: Not sensitizing
Conclusions:
Local lymph node assay (LLNA) of Disodium6-hydroxy-5-[(2-methoxy-4-sulphonato-m-tolyl)azo]naphthalene-2-sulphonate (25956-17-6)was performed on female miceCBA/J was considered to be not sensitizing.
Executive summary:

Local lymph node assay (LLNA) of Disodium6-hydroxy-5-[(2-methoxy-4-sulphonato-m-tolyl)azo]naphthalene-2-sulphonate (25956-17-6) was performed on female miceCBA/J by measuring the cell proliferation in the draining lymph nodes after topical application onto the ears.

In control group only 25µl of vehicle while in test group , 0.5, 1, 2, 4 % (w/v) test substance dissolved in DMSO and 0.5, 1, 2, 4 % in aqua/acetone (1:1) mixed with olive oil (4:1).Positive control p-phenylenediamine at 1 % in DMSO was applied on three consecutive days to the surface of the ear of 5 female CBA/J mice per group. After application, the ears were dried for about 5 minutes by means of a hair dryer.

 

All the animals were observed twice daily for any clinical signs and morbidity/mortality before and after given dose.Body weight was determined on day 1 and 5.On day 5 the mice received an intravenous injection of 250 μl phosphate buffered saline containing 20.8 μCi of [H3] methyl thymidine. After 5hr animals were died by giving CO2-inhalation and the draining auricular lymph node was removed and weighed. The cell suspension for each animal prepared. The cells were predicated by TCA. The liquid scintillation counting as disintegration per minute (dpm) was used to determined radioactivity due to incorporation of [H3] methyl thymidine in the pellets. The stimulation index was calculated by comparing test material with concurrent vehicle control.

 

Result was noted using mean stimulation index ,test substance in concentration 0.5, 1, 2, 4 % (w/v) in DMSO mean stimulation indices of 0.6, 0.7, 0.9 and 0.9 were obtained while test substance as 0.5, 1, 2, 4 % in aqua/acetone (1:1) mixed with olive oil (4:1).The mean stimulation indices of1.2, 1.0, 0.9 and 0.9 were obtained. The positive control (PPD, 1 % in DMSO) caused an increase in the stimulation index by a factor of 7.8.

 

As no relevant increase in the mean stimulation indices was observed (all figures were well below the trigger value of three), Hence it is considered thatDisodium6-hydroxy-5-[(2-methoxy-4-sulphonato-m-tolyl)azo]naphthalene-2-sulphonate (25956-17-6) was not skin sensitizer in mice by usingLocal lymph node assay (LLNA)