N,N'-methylenediacrylamide

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

July 19-27, 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:

EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

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):
- Substance type:
- Physical state: solid
- Analytical purity: No 'Certificate of Analysis' was available
- 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.
- Storage condition of test material: At +10°C to +25°C kept in a cool, dry and well-ventilated place.
- Other:

Specific details on test material used for the study:

Designation N,N‘-Methylenediacrylamide
CAS no. 110-26-9
EC no. 203-750-9
Batch no. LQ27A1677
Receipt no. 60694
Date of receipt May 31, 2016
Characteristics Solid
Stability / Expiry date No data on stability were available to LPT.
Content No 'Certificate of Analysis' was available to LPT.
Storage conditions At +10°C to +25°C kept in a cool, dry and well-ventilated place.
Retention sample of
the test item Stored in LPT’s archives.

Test animals / tissue source

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 32±1°C
- Humidity (%): test was conducted in a bath
- Air changes (per hr): n.a.
- Photoperiod (hrs dark / hrs light): n.a.
EXPERIMENTAL DATE: July 27, 2016

Test system

Vehicle:

physiological saline

Controls:

other: three control corneas

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750 µL
- Concentration (if solution): 20% suspension in 0.9% sodium chloride solution (w/v)

VEHICLE
- Amount(s) applied (volume or weight with unit): 750 µL
- Concentration (if solution): 0.9% sodium chloride solution
- Lot/batch no. (if required): 161418002; B. Braun Melsungen AG, 34212 Melsungen, Germany
- Purity:

Duration of treatment / exposure:

1 day

Observation period (in vivo):

None

Duration of post- treatment incubation (in vitro):

None

Number of animals or in vitro replicates:

3 corneas were used for each treatment group

Details on study design:

Handling and preparation of the bovine eyes
Bovine eyes from cattle in the age range of 6 to 12 months were obtained from a slaughterhouse . To minimize deterioration and bacterial contamination, on collection the eyes were completely submerged in Hanks’ Balanced Salt Solution (HBSS) containing penicillin at 100 IU/mL and streptomycin at 100 µg/mL . Upon arrival at the laboratory, the eyes were examined for defects such as but not limited to increased opacity, scratches, and neovascularisation. Only corneas from eyes free of defects were used.
The quality of each cornea was also evaluated at later steps in the assay. Corneas that had opacity greater than seven opacity units or equivalent for the opacitometer and cornea holders used after an initial one hour equilibration period had to be discarded.
The corneas were dissected with a 2 to 3 mm rim of sclera and mounted in corneal holders with anterior (epithelium) and posterior (endothelium) chambers. Beginning with the posterior chambers, the chambers were filled to excess with pre-warmed Eagle’s Minimum Essential Medium (EMEM) , while preventing bubble formation. The corneal holder was equilibrated at 32±1°C for at least one hour.
After the equilibration period, fresh pre-warmed EMEM was added to both chambers and baseline opacity readings were taken for each cornea. Corneas exhibiting macroscopic tissue damage (e.g. scratches, pigmentation, neovascularisation) or an opacity >7 opacity units were discarded. The mean opacity of all equilibrated corneas was calculated by use of an opacitometer. A minimum of three corneas with opacity values close to the median value for all corneas were selected as negative control corneas. The remaining corneas were then distributed into treatment, solvent and positive control groups.

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

in vitro irritation score

Run / experiment:

1

Value:

-3.88

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation parameter:

in vitro irritation score

Run / experiment:

2

Value:

-5.332

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Results 3

Irritation parameter:

in vitro irritation score

Run / experiment:

3

Value:

-0.582

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Results 4

Irritation parameter:

in vitro irritation score

Run / experiment:

Mean

Value:

-3.265

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

In vivo

Irritant / corrosive response data:

The corneas treated with the negative control item 0.9% sodium chloride solution revealed a mean opacity value of -0.385 ± 0.852 and a mean permeability value of 0.030 ± 0.006. The calculated IVIS value was 0.070 ± 0.899.
The corneas treated with the positive control item 20% Imidazole in 0.9% NaCl solution revealed a mean opacity value of 64.395 ± 6.850 and a mean permeability value of 1.483 ± 0.223 compared to the solvent control. The calculated IVIS value of 86.645 ± 8.731 was within two standard deviations of the current historical mean and well above the cut-off value of 55. Hence, the acceptance criteria for the test were fulfilled.
Following treatment with N,N‘-Methylenediacrylamide a mean opacity of -5.180 ± 2.509 and a mean permeability value of 0.128 ± 0.072 compared to the negative control were determined. The calculated IVIS of -3.265 ± 2.434 is below the cut-off value of 3 (UN GHS no category).

Applicant's summary and conclusion

Interpretation of results:

not irritating

Conclusions:

The test item did not show severely irritant or corrosive properties and consequently

Executive summary:

Under the conditions of thein vitroBCOP test, N,N‘-Methylenediacrylamide, had an IVIS value of <0.01 (-3.265), which is below the cut-off value of 3andconsequently it is not classified as a severe irritant and is not corrosive according to UN GHS classification.