2-amino-5-(2-hydroxyethyl)-6-methyl-4(3H)-pyrimidinone

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Nov 2017 - Mar 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

2-amino-5-(2-hydroxyethyl)-6-methyl-4(3H)-pyrimidinone

Cas Number:

6940-45-0

Molecular formula:

C7H11N3O2

IUPAC Name:

2-amino-5-(2-hydroxyethyl)-6-methyl-4(3H)-pyrimidinone

impurity 1

Reference substance name:

Sodium acetate

EC Number:

204-823-8

EC Name:

Sodium acetate

Cas Number:

127-09-3

Molecular formula:

C2H4O2.Na

IUPAC Name:

sodium acetate

additive 1

Reference substance name:

Silicon dioxide

EC Number:

231-545-4

EC Name:

Silicon dioxide

Cas Number:

7631-86-9

Molecular formula:

SiO2

IUPAC Name:

Silicon dioxide

Test material form:

solid: granular

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 103901
- Expiration date of the lot/batch: 01. Feb. 2019

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room Temperature (20 ± 5°C)
- Stability under test conditions: Stable

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: none

Test animals / tissue source

Species:

human

Details on test animals or tissues and environmental conditions:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 37 ± 1 °C,
- Humidity (%): 80 - 100
-CO2: 5 ± 1 %

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 52.8 an 53.1 mg

Duration of treatment / exposure:

6 hours

Duration of post- treatment incubation (in vitro):

18 hours

Number of animals or in vitro replicates:

2

Details on study design:

- Details of the test procedure used
- RhCE tissue construct used, including batch number : EpiOcularTM tissues were procured from MatTek In Vitro Life Science Laboratories, Mylnské Nivy 73, 82105 Bratislava, Slovakia. Designation of the kit: OCL-200-EIT. Day of delivery: 12. Dec. 2017. Batch no.: 27017
- Doses of test chemical and control substances used : 52.8 an 53.1 mg of test chemical , 50 μL of the controls

Results and discussion

In vitro

Results

Irritation parameter:

other: % cell viability as measured by dehydrogenase conversion of MTT.

Value:

>= 96.4 - <= 102.5

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

other: no eye irritation

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: None

DEMONSTRATION OF TECHNICAL PROFICIENCY: Technical proficiency is demonstrated in Annex 4

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
- Range of historical values if different from the ones specified in the test guideline: no

Any other information on results incl. tables

For the test item and the positive control, the following percentage values of tissue viability were calculated in comparison to the negative control:

Designation	Positive Control	SP02 (trade name: 5OHMlC)
% Viability (Tissue 1)	37.8%	96.4%
% Viability (Tissue 2)	39.8%	102.5%
% Viability Mean	38.8%	99.5%

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of the test, SP02 (trade name: 5OHMlC) is considered non- eye irritant in the EpiOcularTM Eye Irritation Test.

Executive summary:

The test item SP02 (trade name: 5OHMlC) was applied to a three-dimensional human cornea tissue model in duplicate for an exposure time of 6 hours. The solid test item was applied to two tissue replicates. After treatment, the respective substance was rinsed from the tissue; then, cell viability of the tissues was evaluated by addition of MTT, which can be reduced to formazan. The formazan production was evaluated by measuring the optical density (OD) of the resulting solution.

Demineralised water was used as negative control and methyl acetate was used as positive control. The controls showed the following results: After treatment with the negative control, the absorbance values were within the required acceptability criterion of mean OD > 0.8 and <2.5, OD was 1.7. The positive control showed clear eye irritating effects, mean value of the relative tissue viability was 38.8 % (< 50%). Variation within tissue replicates was acceptable (< 20%). After treatment with the test item, the mean value of relative tissue viability was 99.5 %. This value is above the threshold for eye irritation potential (≤ 60%).

Under the conditions of the test, SP02 (trade name: 5OHMlC) is considered non- eye irritant in the EpiOcularTM Eye Irritation Test.