(3-methacrylamidopropyl)trimethylammonium chloride

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2007-01-16 to 2007-02-06

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study. 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)

Version / remarks:

adopted 24 April 2002

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:

(3-methacrylamidopropyl)trimethylammonium chloride

EC Number:

257-182-1

EC Name:

(3-methacrylamidopropyl)trimethylammonium chloride

Cas Number:

51410-72-1

Molecular formula:

C10 H21 N2 O. Cl

IUPAC Name:

trimethyl-[3-(2-methylprop-2-enoylamino)propyl]azanium chloride

Test material form:

other: aqueous solution

Details on test material:

- Name of test material (as cited in study report): 1-Propanaminium, N,N,N-trimethyl-3-[(2-methyl-1-oxo-2-propenyl)amino]-, chlorid

In vivo test system

Test animals

Species:

mouse

Strain:

other: CBA/CaOlaHsd

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Age at study initiation: 7-8 weeks (beginning of acclimatisation)
- Weight at study initiation: 18.4 g ±0.7
- Housing: single, Makrolon Type I, with wire mesh top (EHRET GmbH, D-79302 Emmendingen), granulated soft wood bedding (Harlan Winkelmann GmbH, D-33178 Borchen)
- Diet (e.g. ad libitum): pelleted standard diet, ad libitum (Harlan Winkelmann GmbH, D-33178 Borchen)
- Water (e.g. ad libitum): tap water, ad libitum (Gemeindewerke, D-64380 Rossdorf)
- Acclimation period: not given

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3°C
- Humidity (%): 30-72%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES (GLP Main Experiment): From: 2007-01-31 To: 2007-02-06

Study design: in vivo (LLNA)

Vehicle:

dimethylformamide

Concentration:

- non-GLP Pretest (test substance): 12.5% (w/v), 25% (w/v), 50%(w/v) and 100% (w/v)
- Main study (test substance): 25% (w/v); 50% (w/v) and 100% (w/v)
- Positive control: α-Hexylcinnamaldehyde in acetone: olive oil (4+1) at conc. 5% (w/v), 10% (w/v) and 25% (w/v)
- Negative control: equivalent volume of vehicle (dimethylformamide)

No. of animals per dose:

4

Details on study design:

RANGE FINDING TESTS:
- Irritation: In a non-GLP conform pre-test in two mice, test item concentrations of 12.5%, 15%, 50% and 100% were tested on one ear each. No irritation effects were observed at these concentrations after a single application.

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: LLNA (OECD 429, 24 April 2002)
- Criteria used to consider a positive response: The test item is regarded as a sensitizer in the LLNA if the exposure to one or more test concentration resulted in a 3-fold or greater increase in incorporation of 3HTdR compared with concurrent controls, as indicated by the Stimulation Index (S.I.).

TREATMENT PREPARATION AND ADMINISTRATION:
Application: topical (epidermal) application to the dorsal surface of each ear lobe (left and right) once daily on three consecutive days with different test item concentrations of 25, 50 and 100% (w/v) in dimethylformamide. The application volume (25 µL) was spread over the entire dorsal surface (approx. diameter of 8 mm). The control animals were treated with an equivalent volume of vehicle alone.
Administration of 3H-Methyl Thymidine: 250 µL of 81.0 µCi/mL 3HTdR (corresponds to 20.3 µCi 3HTdR per mouse) by intravenous injection via a tail vein five days after the first topical application.
Determination of incorporated 3HTdR: All mice were euthanized approx. 5 hours after treatment with 3HTdR. The draining lymph nodes were excised and pooled per group (8 nodes per group). Single cell suspension of pooled lymph node cells were prepared. After resuspension in 5% trichloroacetic acid and mixing with scintillation liquid, the level of 3HTdR incorporation was measured on a β-scintillation counter. Background 3HTdR levels were measured in two 1 mL-aliquots of 5% trichloroacetic acid.
The proliverative response of lymph node cells is expressed as disintegrations per minute per lymph node (DPM/node) and as the ratio of 3HTdR incorporated into lymph node cells relative to that recoreded for control lymph nodes (stimulation index).

Positive control substance(s):

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

Results and discussion

Positive control results:

The positive control test item resulted in a stimulation index of 2.04 at 5% (w/v) and S.I.=6.31 at 10%, respectively. Accordingly, the EC3 was calculated to 6.1% and thus indicated the adequate function of the test system.

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: Test substance: 25% (w/v): S.I. = 0.69 50% (w/v): S.I. = 0.56 100% (w/v): S.I. = 0.59 The EC3 value could not be calculated, since all S.I.s are below 3.

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: Control group: 407.8 DPM/lymph node (3262.0 total DPM) Test substance: 25% (w/v): 280.6 DPM/lymph node (2245.0 total DPM) 50% (w/v): 229.0 DPM/lymph node (1832.2 total DPM) 100% (w/v): 239.3 DPM/lymph node (1914.6 total DPM)

Any other information on results incl. tables

MAIN STUDY

No deaths, no symptoms of local toxicity at the ears of the animals and no systemic findings were observed during the study period. Body weights of the test animals, which were recorded prior to the first application and prior to treatment with ³HTdR, were within the range commonly recorded for animals of this strain and age (only three mice lost 0.1-1.7 g of their initial weight).

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: other: CLP, EU GHS (Regulation (EC) No 1272/2008)

Conclusions:

In this Local Lymph Node Assay, MAPTAC (50% solution in water) was not identified to be a dermal sensitiser.

Executive summary:

In a dermal sensitization study with the test substance MAPTAC (49.8% solution in water) in Dimethylformamide, groups of 4 female CBA/CaOlaHsd mice were tested using the LLNA method according to the OECD Guideline 429, 24 April 2002. Proliferation response of lymph node cells was measured by DPM counting of incorporated 3HTdR.

A substance is regarded as dermally sensitising if the S.I. ≥ 3.

STIMULATION INDICES (S.I.) of 0.69, 0.56 and 0.59 were determined for the test substance at concentrations of 25%, 50% and 100%, respectively in Dimethylformamide (w/v). The positive control (α-Hexylcinnamaldehyde) gave the appropriate response (EC3=6.1% in acetone:olive oil (4+1, w/v)).

In this study, MAPTAC (49.8% solution in water) is not a dermal sensitiser.

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