Bis(4-hydroxy-N-methylanilinium) sulphate

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data from peer-reviewed journal

Data source

Reference

Reference 1

Reference Type:

publication

Title:

Comparison of the local lymph node assay with the guinea pig maximization test for the detection of a range of contact allergens

Author:

D.A. Basketter and E.W. Scholes

Year:

1992

Bibliographic source:

Fd. Chem. Toxic .Vol. 30, No 1, pp.65-69,1992

Reference 2

Reference Type:

publication

Title:

Compilation of Historical local Lymph Node Data for Evaluation of skin Sensitization Alternative method

Author:

G. Frank Gerberick, Cindy A. Ryan, Petra S. Kern, Harald Schlatter, Rebecca J. Dearman, Ian Kimber, Grace Y. Patlewicz, and David A. Basketter

Year:

2005

Bibliographic source:

Dermatitis, Vol 16, No 4, pp 157-202, 2005

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

Skin sensitization study of N-methyl-p-aminophenol sulphate was performed in mouse by LLNA method

GLP compliance:

not specified

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

Bis(4-hydroxy-N-methylanilinium) sulphate

EC Number:

200-237-1

EC Name:

Bis(4-hydroxy-N-methylanilinium) sulphate

Cas Number:

55-55-0

Molecular formula:

C14H20N2O6S

IUPAC Name:

bis(4-hydroxy-N-methylanilinium) sulfate

Test material form:

solid

Details on test material:

- Name of test material: p-Methylaminophenol sulfate- IUPAC name: Bis(4-hydroxy-N-methylanilinium) sulphate- Molecular formula: C14H20N2O6S- Molecular weight: 344.386 g/mole- Smiles:CNc1ccc(cc1)O.CNc1ccc(cc1)O.OS(=O)(=O)O- Inchl: 1S/2C7H9NO.H2O4S/c2*1-8-6-2-4-7(9)5-3-6;1-5(2,3)4/h2*2-5,8-9H,1H3;(H2,1,2,3,4)- Substance type: Organic- Physical state: Solid crystalline (off white - white)

Specific details on test material used for the study:

- Name of test material: p-Methylaminophenol sulfate- IUPAC name: Bis(4-hydroxy-N-methylanilinium) sulphate- Molecular formula: C14H20N2O6S- Molecular weight: 344.386 g/mole- Smiles:CNc1ccc(cc1)O.CNc1ccc(cc1)O.OS(=O)(=O)O- Inchl: 1S/2C7H9NO.H2O4S/c2*1-8-6-2-4-7(9)5-3-6;1-5(2,3)4/h2*2-5,8-9H,1H3;(H2,1,2,3,4)- Substance type: Organic- Physical state: Solid crystalline (off white - white)

In vivo test system

Test animals

Species:

mouse

Strain:

CBA/Ca

Sex:

male/female

Details on test animals and environmental conditions:

Details on test animalTEST ANIMALS- Source: No data available- Age at study initiation:8-12 weeks - Weight at study initiation: No data available- Housing: No data available- Diet (e.g. ad libitum): No data available- Water (e.g. ad libitum): No data available- Acclimation period: No data availableENVIRONMENTAL CONDITIONS- Temperature (°C): No data available- Humidity (%):No data available- Air changes (per hr): No data available- Photoperiod (hrs dark / hrs light): No data available

Study design: in vivo (LLNA)

Vehicle:

dimethylformamide

Concentration:

0.5, 1, 2.5, %

No. of animals per dose:

Total:160.5%: 41% : 42.5%: 4Vehicle control:4

Details on study design:

Details on study designPRE-SCREEN TESTS:- Compound solubility: Test material soluble in DMF- Irritation: No data available- Systemic toxicity:- Ear thickness measurements: 4-5 days after the first application, all mice injected intravenously through the tail vein with 250 μl of phosphate buffered saline (PBS) containing 20 μCi of tritiated thymidine (³H-TdR). After approximately 5 hours, they were killed by CO2 asphyxiation and the auricular lymph nodes excised. The nodes from each group were pooled, a suspension of auricular lymph node cells prepared, and proliferation of these cells measured using β-scintillation counting. The results were used to calculate the Stimulation Index (SI) for proliferation. The EC3 value (the theoretical concentration resulting in an SI value of 3) was subsequently determined.- Erythema scores: No data available MAIN STUDYANIMAL ASSIGNMENT AND TREATMENT- Name of test method: LLNA- Criteria used to consider a positive response: A chemical was regarded as a sensitizer in the LLNA if at least one concentration of the chemical resulted in a threefold or greater increase in HTdR incorporation compared with control value.TREATMENT PREPARATION AND ADMINISTRATION: The dose volume of 25 μl was applied to the dorsal surface of both ears at concentrations of 0.5%, 1%, 2.5% once daily for 3 consecutive days. Control animals received Vehicle alone.

Positive control substance(s):

not specified

Statistics:

No data available

Results and discussion

Positive control results:

No data available

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Value:

2.5

Test group / Remarks:

0.5% test group

Remarks on result:

other: positive effect was observed

Results 2

Parameter:

SI

Value:

3.4

Test group / Remarks:

1% test group

Remarks on result:

other: positive effect was observed

Results 3

Parameter:

SI

Value:

6.7

Test group / Remarks:

2.5% test group

Remarks on result:

other: positive effect was observed

Cellular proliferation data / Observations:

The ratio of test to control lymphocyte proliferation (T/C) was determined as 2.5, 3.4 and 6.7 indcated positive effect for skin sensitization.

Applicant's summary and conclusion

Interpretation of results:

other: sensitizing

Conclusions:

The N-methyl p-aminophenol sulphate (55-55-0) was considered to be sensitizing in mice by LLNA method

Executive summary:

The skin sensitization studyN-methyl p-aminophenol sulphate(55-55-0)was performed on 16CBA/Ca male or female mice of age 8-12 weeks .The test material soluble in DMF.The dose volume of 25 μl was applied to the dorsal surface of both ears at concentrations of 0.5%, 1%, and 2.5% once daily for 3 consecutive days. Control animals received Vehicle alone.4-5 days after the first application, all mice injected intravenously through the tail vein with 250 μl of phosphate buffered saline (PBS) containing 20 μCi of tritiated thymidine (³H-TdR). After approximately 5 hours, they were killed by CO2 asphyxiation and the auricular lymph nodes excised. The nodes from each group were pooled, a suspension of auricular lymph node cells prepared, and proliferation of these cells measured using β-scintillation counting. The results were used to calculate the Stimulation Index (SI) for proliferation. The EC3 value (the theoretical concentration resulting in an SI value of 3) was subsequently determined.A chemical was regarded as a sensitizer in the LLNA if at least one concentration of the chemical resulted in a threefold or greater increase in HTdR incorporation compared with control value. The ratio of test to control lymphocyte proliferation (T/C) was determined as 2.5, 3.4 and 6.7 HenceN-methyl p-aminophenol sulphate(55-55-0) was considered to be sensitizing in mice by LLNA method.