N,N'-methylenediacrylamide

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

July 19 to August 30, 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 442B (Skin Sensitization: Local Lymph Node Assay: BrdU-ELISA)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EC method B.51: Skin Sensitisation, Local Lymph Node Assay: BrdU-ELISA, Commission Regulations (EU) No. 640/2012, adopted July 06, 2012.

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:

N,N'-methylenediacrylamide

EC Number:

203-750-9

EC Name:

N,N'-methylenediacrylamide

Cas Number:

110-26-9

Molecular formula:

C7H10N2O2

IUPAC Name:

N,N'-methylenediacrylamide

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): N,N‘-Methylenediacrylamide
- Molecular formula (if other than submission substance):
- Molecular weight (if other than submission substance):
- Smiles notation (if other than submission substance):
- InChl (if other than submission substance):
- Structural formula attached as image file (if other than submission substance): see Fig.
- Substance type:
- Physical state:
- Analytical purity:
- Impurities (identity and concentrations):
- Composition of test material, percentage of components:
- Isomers composition:
- Purity test date:
- Lot/batch No.: LQ27A1677
- Expiration date of the lot/batch:
- Radiochemical purity (if radiolabelling):
- Specific activity (if radiolabelling):
- Locations of the label (if radiolabelling):
- Expiration date of radiochemical substance (if radiolabelling):
- Stability under test conditions: No data on stability were available to LPT.
- Storage condition of test material: No data on stability were available.
- Other: No 'Certificate of Analysis' was available .

In vivo test system

Test animals

Species:

mouse

Strain:

CBA:JN

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: JANVIER LABS.
Route des Chènes Secs C.S.4105
Le Genest-Saint-Isle
53941 Saint Berthevin Cedex
France

- Age at study initiation: approx. 12 weeks
- Weight at study initiation: 20 - 27 g
- Housing: Before application the animals were housed in groups of 5 animals in MAKROLON cages (type III) with a basal surface of approx. 39 cm x 23 cm and a height of approx. 15 cm. After application the animals were housed singly in order to prevent them licking off the test item from the ears of the other animals.
- Diet (e.g. ad libitum): ssniff® R/M-H V1534 (ssniff Spezialdiäten GmbH).This food was offered ad libitum.
- Water (e.g. ad libitum): Tap water was offered ad libitum.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): room temperature of 22°C ± 3°C (maximum range)
- Humidity (%): relative humidity of 55% ± 15% (maximum range)
- Air changes (per hr): 15 to 20 times per hr
- Photoperiod (hrs dark / hrs light): The rooms were alternately lit (about 150 lux at approx. 1.50 m room height) and darkened for periods of 12 hours each.

IN-LIFE DATES: From: To: First administration August 25, 2016; Termination of the experimental phase August 30, 2016

Study design: in vivo (LLNA)

Vehicle:

dimethylformamide

Concentration:

10% (w/w)
25% (w/w)
50% (w/w)

No. of animals per dose:

30 (6 groups of 5 animals each)

Details on study design:

RANGE FINDING TESTS:
- Compound solubility: 50% in N,N-dimethylformamide (w/w)
- Irritation: none
- Lymph node proliferation response: none

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: LOCAL LYMPH NODE ASSAY: BRDU-ELISA
- Criteria used to consider a positive response: An index for the lymph node cell count ≥ 1.6 is considered positive.

TREATMENT PREPARATION AND ADMINISTRATION:
An index for the lymph node cell count ≥ 1.6 is considered positive.
The test item formulations were administered to the dorsum of both animal's ears at an application volume of 25 µL/ear.

Positive control substance(s):

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

Statistics:

As no concentration-related increased values were observed, no statistical evaluation was carried out. Comparison of positive control to its negative control was carried out using Student’s t-test.

Results and discussion

Positive control results:

Treatment with the positive control item caused the expected increases in the BrdU labelling index. Therefore, the study can be regarded as valid.

In vivo (LLNA)

Results

Results 1

Key result

Parameter:

SI

Value:

0.837

Test group / Remarks:

Group 2 (10%) / BrdU labelling index

Results 2

Key result

Parameter:

SI

Value:

1.019

Test group / Remarks:

Group 2 (10%) / Ear weight

Results 3

Key result

Parameter:

SI

Value:

1.038

Test group / Remarks:

Group 2 (10%) / Difference of ear thickness (TD 3)

Results 4

Key result

Parameter:

SI

Value:

1.052

Test group / Remarks:

Group 2 (10%) / Difference of ear thickness (TD 6)

Results 5

Key result

Parameter:

SI

Value:

0.771

Test group / Remarks:

Group 3 (25%) / BrdU labelling index

Results 6

Key result

Parameter:

SI

Value:

1.081

Test group / Remarks:

Group 3 (25%) / Ear weight

Results 7

Key result

Parameter:

SI

Value:

1.043

Test group / Remarks:

Group 3 (25%) / Difference of ear thickness (TD 3)

Results 8

Key result

Parameter:

SI

Value:

1.033

Test group / Remarks:

Group 3 (25%) / Difference of ear thickness (TD 6)

Results 9

Key result

Parameter:

SI

Value:

0.743

Test group / Remarks:

Group 4 (50%) / BrdU labelling index

Results 10

Key result

Parameter:

SI

Value:

1.063

Test group / Remarks:

Group 4 (50%) / Ear weight

Results 11

Key result

Parameter:

SI

Value:

1.071

Test group / Remarks:

Group 4 (50%) / Difference of ear thickness (TD 3)

Results 12

Key result

Parameter:

SI

Value:

1.038

Test group / Remarks:

Group 4 (50%) / Difference of ear thickness (TD 6)

Any other information on results incl. tables

Study results: stimulation indices (SI):

Parameter	Group 1, negative control	Group 2, test item 10%	Group 3, test item 25%	Group 4, test item 50%	Group 5, positive control	Group 6, vehicle of positive control
BrdU labelling index	1.000	0.837	0.771	0.743	1.778	1.000
Ear weight	1.000	1.019	1.081	1.063	1.228	1.000
Difference of ear thickness (TD 3)	1.000	1.038	1.043	1.071	1.078	1.000
Difference of ear thickness (TD 6)	1.000	1.052	1.033	1.038	1.128	1.000

       ____ increases significantly compared to control at p ≤ 0.01

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, N,N‘-Methylenediacrylamide did not reveal any skin sensitising properties in the local lymph node assay: BrdU-ELISA.