2,2'-iminobis[4,6-diamino-1,3,5-triazine]

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

06-Apr-2011 to 11-Apr-2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: EU method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: OECD Guideline no. 439: In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Melam

IUPAC Name:

Melam

Constituent 2

Reference substance name:

2,2'-iminobis[4,6-diamino-1,3,5-triazine]

EC Number:

222-695-1

EC Name:

2,2'-iminobis[4,6-diamino-1,3,5-triazine]

Cas Number:

3576-88-3

Molecular formula:

C6H9N11

IUPAC Name:

N~2~-(4,6-diamino-1,3,5-triazin-2-yl)-1,3,5-triazine-2,4,6-triamine

Details on test material:

- Name of test material (as cited in study report): Melam
- CAS: 3576-88-3
- Substance type: White powder
- Physical state: Solid
- Storage condition of test material: At room temperature in the dark

Test animals

Species:

other: in vitro study

Test system

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 10 mg, moistened with 5 µl water

NEGATIVE CONTOL:
- Amount(s) applied (volume or weight with unit): 10 µl Phosphate buffered saline

POSITIVE CONTROL
- Amount(s) applied (volume or weight with unit): 10 µl
- Concentration (if solution): 5% (aq) Sodium dodecyl sulphate

Duration of treatment / exposure:

Exposure:15 minutes
Post incubation period: 42 hours

Details on study design:

TEST SITE
- Area of exposure: human epidermis model
- % coverage: 0.38 cm2

REMOVAL OF TEST SUBSTANCE
- Washing (if done): phosphate buffered saline
- Time after start of exposure: 15 minutes

POST INCUBATION PERIOD
- 42 hours

SCORING SYSTEM:
- After a 42 hour incubation period, determination of the cytotoxic (irritancy) effect was performed. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT at the end of the treatment. Cell viability was calculated for each tissue as a percentage of the mean of the negative control tissues.

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

other: other: percentage viability

Value:

104

Remarks on result:

other:

Remarks:

Basis: other: percentage of control. Time point: 15 minutes. Remarks: test substance. (migrated information)

Results 2

Irritation / corrosion parameter:

other: other: percentage viability

Value:

6

Remarks on result:

other:

Remarks:

Basis: other: percentage of control. Time point: 15 minutes. Remarks: Positive control. (migrated information)

In vivo

Other effects:

Melam was checked for possible direct MTT reduction by adding the test substance to MTT medium. Because no colour change was observed it was concluded that Melam did not interact with MTT.

Any other information on results incl. tables

Skin irritation is expressed as the remaining cell viability after exposure to the test substance. The relative mean tissue viability obtained after 15 minutes treatment with Melam compared to the negative control tissues was 104%.

Since the mean relative tissue viability for Melam was above 50% Melam is considered to be non-irritant.

The positive control had a mean cell viability after 15 minutes exposure of 6%. The absolute mean OD570 of the negative control tissues was within the laboratory historical control data range. The standard deviation value of the percentage viability of three tissues treated identically was less than 4 %, indicating that the test system functioned properly.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

It is concluded that this test is valid and that Melam, CAS 3576-88-3, is non-irritant in the in vitro skin irritation test.

Executive summary:

Skin irritation is expressed as the remaining cell viability after exposure to the test substance. The relative mean tissue viability obtained after 15 minutes treatment with Melam compared to the negative control tissues was 104 %. Since the mean relative tissue viability for Melam was above 50 % after 15 minutes treatment Melam is considered to be non-irritant.

 

The positive control had a mean cell viability of 6 % after 15 minutes exposure. The absolute mean OD₅₇₀(optical density at 570 nm) of the negative control tissues was within the laboratory historical control data range. The standard deviation value of the percentage viability of three tissues treated identically was less than 4 %, indicating that the test system functioned properly.