5-nitro-2,4,6-triaminopyrimidine

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2014-2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant study that is suitable for classification and labelling

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD 439 and OECD 431

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

5-nitro-2,4,6-triaminopyrimidine

EC Number:

213-742-7

EC Name:

5-nitro-2,4,6-triaminopyrimidine

Cas Number:

1006-23-1

Molecular formula:

C4 H6 N6 O

IUPAC Name:

5-nitropyrimidine-2,4,6-triamine

Details on test material:

-Name in report: 5-Nitroso-2,4,6-triaminopyrimidine
- Physical state: Solid (pink to red)
- Analytical purity: 100% (HPLC)
- Water content: 0.2%
- Lot/batch No. 1101-1217
- Expiry date: 10 Oct 2023
- Stability under test conditions: The stability of the test item under storage conditions over the study period was guaranteed by the sponsor.
- Storage condition of test material: Room temperature
- Homogeneity: The test substance was homogeneous by visual inspection.

Test animals

Species:

human

Strain:

other: not applicable

Details on test animals or test system and environmental conditions:

EPIDERM(TM) 200 kit from MaTek Corp, Ashland MA, USA
reconstructed epidermis, surface 0.6cm2 and cultured in vessels of 1 cm diameter.

Corrosion test:
four tissues are treated with each test substance, the PC and NC, respectively. Two of these 4 tissues are required for the 3-minute exposure and 2 pieces are required for the 1-hour exposure.

Irritation test: three tissues are treated with each test substance the PC and NC 1 1 respectively.

Test system

Type of coverage:

open

Preparation of test site:

other: not applicale

Vehicle:

unchanged (no vehicle)

Controls:

other: untreated skin tissue

Amount / concentration applied:

corrosion test: 25 μL de-ionized water is applied first. Thereafter, a bulk volume of 25 μL of the test
material is applied with a sharp spoon and homogeneously distributed together with the water.

Irritation test: 25 μL sterile PBS is applied first. Thereafter, a bulk volume of 25 μL of the test material is applied with a sharp spoon and homogeneously distributed together with the fluid.

Duration of treatment / exposure:

3 min, 60 min

Observation period:

48h (recovery period, including 3h MTT incubation)

Number of animals:

not applicable

Details on study design:

To determine whether the test substance is able to reduce MTT directly, the test substance was incubated with the substrate.
Three tissues were treated with each test substance, positive and negative controls, espectively.

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

other: other: viability of reconstituted epidermis (corrostion test)

Value:

95.5

Remarks on result:

other:

Remarks:

Basis: mean. Time point: 60 min. Max. score: 100.0. Reversibility: other: not applicable. Remarks: Score given in %. (migrated information)

Results 2

Irritation / corrosion parameter:

other: other: viability of reconstituted epidermis (irritation test)

Value:

98.1

Remarks on result:

other:

Remarks:

Basis: mean. Time point: 60 min. Max. score: 100.0. Reversibility: other: not applicable. Remarks: Score given in %. (migrated information)

Any other information on results incl. tables

Table: Corrosion test - exposure period 1 h: Individual and mean OD570 values, individual and mean viability values,

standard deviations and coefficient of variation

			tissue 1	tissue 2	mean	SD	CV (%)
Negative control	viable tissues	mean OD570	2.138	2.122	2.130	0.011
		viability [% of NC]	100.4	99.6	100.0	0.5	0.5
	killed tissues	mean OD570	0.103	0.114	0.109	0.007
		viability [% of NC]	4.8	5.3	5.1	0.3
Test substance	viable tissues	mean OD570	2.122	1.980	2.051	0.100
		viability [% of NC]	99.6	93.0	96.3	4.7	4.9
	killed tissues	mean OD570KC NC corrected	0.004	0.030	0.017	0.018
		viability [% of NC]	0.2	1.4	0.8	0.9
	Final mean viability of tissues after correction of killed tissues [% of NC]:				95.5
Positive control  5% (w/v) sodium dodecyl sulfate	viable tissues	mean OD570	0.131	0.152	0.141	0.015
		viability [% of NC]	6.1	7.1	6.6	0.7	10.5

Table: Irritation test - Individual and mean OD570 values, individual and mean viability values, standard deviations and

coefficient of variation

				tissue 1		tissue 2		tissue 3		mean		SD               CV [%]
negative control	viable tissues	mean OD570		2.304		2.649		2.488		2.480		0.173
		viability [% of NC]		92.9		106.8		100.3		100.0		7.0	7.0
	killed cell tissues	mean OD570		0.062		0.066		0.055		0.061		0.005
		viability [% of NC]		2.5		2.6		2.2		2.4		0.2
Test substance	viable tissues	mean OD570		2.485		2.357		2.472		2.438		0.071
		viability [% of NC]		100.2		95.0		99.6		98.3		2.8	2.9
	killed cell tissues*	mean OD570KC NC corrected		0.010		0.002		0.000		0.004		0.005
		viability [% of NC]		0.4		0.1		0.0		0.2		0.2
		Final mean viability of tissues after killed cell correction [% of NC]:								98.1
Positive control	viable tissues	mean OD570		0.063		0.068		0.072		0.067		0.005
		viability [% of NC]		2.5		2.7		2..9		2.7		0.2	7.1

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU