Ammonium 4-acetoacetylamino-5-methoxy-2-methylbenzenesulphonate

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: well performed OECD study with GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

Ammonium 4-acetoacetylamino-5-methoxy-2-methylbenzenesulphonate

EC Number:

276-769-3

EC Name:

Ammonium 4-acetoacetylamino-5-methoxy-2-methylbenzenesulphonate

Cas Number:

72705-22-7

Molecular formula:

C12H15NO6S.H3N

IUPAC Name:

ammonium 4-acetoacetylamino-5-methoxy-2-methylbenzenesulphonate

Details on test material:

purity: 95.6 % (contains residual water)

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Strain: CBA/CaOlaHsd
- Source: Harlan Netherlands, B.V. Postbus 6174, NL - 5960 AD Horst / The Netherlands
- Age at study initiation: 7 - 8 weeks (beginning of acclimatisation)
- Weight at study initiation: 18.6 - 22.1 g
- Housing: single in Makrolon Type I cages, with wire mesh top
- Diet: pelleted standard diet, ad libitum (Harlan Winkelmann GmbH, D-33178 Borchen)
- Water: tap water, ad libitum, (Gemeindewerke, D-64380 Rossdorf)

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+-3
- Humidity (%): 30 - 86
- Photoperiod: artificial light 6.00 a.m. - 6.00 p.m.

Study design: in vivo (LLNA)

Vehicle:

dimethyl sulphoxide

Concentration:

0 %, 6.25 %, 12.5 %, 25 %

No. of animals per dose:

4

Details on study design:

Observations:
Mortality / Viability: once daily (week day) from experimental start to necropsy.
Body weights: prior to the first application and prior to treatment with 3HTdR.
Clinical signs (local / systemic): once daily (week day). Especially the treatment sites were observed carefully.

Positive control substance(s):

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

Results and discussion

Positive control results:

Positive control substance: alpha-Hexylcinnamaldehyde
Vehicle: acetone:olive oil (4+1)
concentrations: 5 %, 10 % , 25 %
S.I . 2.04 6.31 12.45

EC3-value: 6.1

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: see Remark

Remarks:

Test item Measurement Calculation concentration Group DPM DPM-BG number of DPM per Result % (w/v) lymph nodes lymph node S.I. --- BG I 15.43 --- --- --- --- --- BG II 16.47 --- --- --- --- 0 1 6052.97 6037.0 8 754.6 --- 6.25 2 7270.68 7254.7 8 906.8 1.20 12.5 3 11548.60 11532.7 8 1441.6 1.91 25 4 10917.00 10901.1 8 1362.6 1.81 BG = Background The EC3 Value could not be calculated, since all SI´s are below 3.

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: see below

Any other information on results incl. tables

No deaths occurred during the study period.

No symptoms of local toxicity at the ears of the animals and no systemic findings were observed during the study period.

The body weight of the animals was within the range commonly recorded for animals of this strain and age.

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test item was not a skin sensitiser under the described conditions.

Executive summary:

In order to study a possible contact allergenic potential a Local Lymph Node Assay (OECD 429) was performed with the substance.Three groups each of four female mice were treated daily with the test item at concentrations of 6.25, 12.5, and 25% (w/v) in DMSO by topical application to the dorsum of each ear lobe (left and right) for three consecutive days. A control group of four mice was treated with the vehicle (DMSO) only. Five days after the first topical application the mice were injected intravenously into a tail vein with radio-labelled thymidine (3H-methyl thymidine). Approximately five hours after intravenous injection, the mice were sacrificed, the draining auricular lymph nodes excised and pooled per group. Single cell suspensions of lymph node cells were prepared from pooled lymph nodes which were subsequently washed and incubated with trichloroacetic acid overnight. The proliferative capacity of pooled lymph node cells was determined by the incorporation of 3H-methyl thymidine measured in a ß-scintillation counter.

All treated animals survived the scheduled study period and no signs of toxicity were observed.

In this study Stimulation Indices of 1.20, 1.91, and 1.81 were determined with the test item at concentrations of 6.25, 12.5, and 25% in DMSO. The EC3 value could not be calculated, since none of the tested concentrations induced an S.I. greater than 3.