5-bromo-4-chloro-6-cyclopropylpyrimidine

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Nov. 2016 - Mar. 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

The study was conducted to meet the known requirements of Method B46 of Council Regulation (EC) No 761/2009 and OECD Guidelines for Testing of Chemicals Method 439 (adopted 28 July 2015).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Remarks:

given by compliance statement with signature

Test material

Test material information

Constituent 1

Reference substance name:

5-bromo-4-chloro-6-cyclopropylpyrimidine

EC Number:

823-376-5

Cas Number:

27192-21-8

Molecular formula:

C7H6BrClN2

IUPAC Name:

5-bromo-4-chloro-6-cyclopropylpyrimidine

Test material form:

solid: particulate/powder

In vitro test system

Test system:

human skin model

Remarks:

skin model EpiDermTM SIT (EPI-200)

Cell type:

non-transformed keratinocytes

Vehicle:

other: deionised water and isopropanol

Details on test system:

EpiDermTM SIT (EPI-200) tissues were kept in their packaging until the next step. The tissues were set up the day prior to treatment by placing each tissue onto 0.9 mL maintenance medium (supplied with the EpiDermTM SIT (EPI-200) tissues) in 6-well plates and incubating at 37°C.

Experiment 1:On a total of three tissues per test article, negative and positive
control. A further test was conducted on a total of 6 tissues per test article, and three per
negative and positive control.
25 mg of the test article was added topically to the tissues. In the first experiment the tissues
were moistened with 25 μL of PBS, prior to application. In the second experiment an extra
treatment condition was included, omitting the 25 μL of PBS prior to application. A volume
of 30 μL of the positive and negative control solutions was used.
In the first experiment the treated tissues were paced into an incubator at 37±1ºC, 5±1% CO2
for 35 minutes. The plates were removed from the incubator and placed into a sterile hood
until the 60 minute treatment period was complete for each tissue. Following treatment,
substances were removed by washing the tissues. The tissues were then placed on the
appropriate medium and incubated for 41 hours and 50 minutes.

Experiment 2:
The treated tissues were paced into an incubator at 37±1ºC, 5±1%
CO2 for 35 minutes. The plates were removed from the incubator and placed into a sterile
hood until the 60 minute treatment period was complete for each tissue. Following treatment,
substances were removed by washing the tissues. The tissues were then placed on the
appropriate medium and incubated for 40 hours and 15 minutes

Control samples:

yes, concurrent negative control

yes, concurrent positive control

yes, concurrent MTT non-specific colour control

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

1

Value:

62.6

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

100 %

Positive controls validity:

valid

Remarks:

3.3 %

Remarks on result:

other:

Remarks:

The standard deviation for test article treated tissues was >18%. Therefore this assay acceptance criterion was not met.

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

2

Value:

8.8

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

100 %

Positive controls validity:

valid

Remarks:

2.8 %

Remarks on result:

other: Result is for not moistened tissues

Remarks:

The standard deviation for pre-moistened, test article treated tissues was >18%. Therefore this assay acceptance criterion was not met. The assay acceptance criterion was met for those tissues that were not moistened.

Applicant's summary and conclusion

Interpretation of results:

other: considered to be irritant

Conclusions:

The test article, IN 79479, was considered to be irritant in the in vitro skin model EpiDermTM
As no study is available for corrosion and the EpiDermTM can't differ between corrosion/irritation, IN 79479 was classified with H314.