5-Diazo-2,4,6(1H, 3H, 5H)-pyrimidinetrione

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: according to guideline and GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD TG 431 and EC B40

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

5-Diazo-2,4,6(1H, 3H, 5H)-pyrimidinetrione

EC Number:

608-591-2

Cas Number:

31221-06-4

Molecular formula:

C4H2N4O3

IUPAC Name:

5-Diazo-2,4,6(1H, 3H, 5H)-pyrimidinetrione

Details on test material:

content 96.7 %

Test animals

Species:

other: in vitro

Strain:

other: in vitro

Details on test animals or test system and environmental conditions:

in vitro test

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: in vitro test: negative control 0.9 % NaCl

Amount / concentration applied:

25 mg

Duration of treatment / exposure:

3 min and 60 min (3 inserts per period)

Observation period:

after the exposure period of 3 min or 60 min the inserts were washed carefully in PBS and MTT reduction assay was performed

Number of animals:

in vitro test

Details on study design:

The experiment was carried out in accordance with tje OECD 431 guideline as well as with an EC guideline (amending Council Directtive 67/548/EEC B 40 skin corrosion) using commercially available reconstructed human epidermis(RHS) model EST-1000. Undiluted diazobarbitursäure was applied topically to the RHS model: 25 mg of the test item per insert ( plus 30 µl 0.9 % NaCl to moisten and ensure good contact wtih the skin; RT: three inserts per period of incubation). After the exposure period of 3 min or 60 min the inserts were washed carefully in PBS. Afterwards the MTT reduction assay was performed to measure the cell viability.

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

other: other: absorption: at 570 nm

Value:

2.27

Remarks on result:

other:

Remarks:

Basis: other: negative control. Time point: 3 min. Max. score: 2.27. Reversibility: no data. Remarks: Optical Density (OD): 2.27 = 100 % viability. (migrated information)

Results 2

Irritation / corrosion parameter:

other: other: absorption at 570 nm

Value:

2.3

Remarks on result:

other:

Remarks:

Basis: other: test item. Time point: 3 min. Max. score: 2.27. Reversibility: no data. Remarks: OD mean 2.27 = 100 % viability; OD mean 2.30 = 101 % viability. (migrated information)

Results 3

Irritation / corrosion parameter:

other: other: absorption at 570 nm

Value:

2.27

Remarks on result:

other:

Remarks:

Basis: other: negative control. Time point: 60 min . Max. score: 2.27. Reversibility: no data. Remarks: Optical Density (OD):2.27 = 100 % viability; . (migrated information)

Results 4

Irritation / corrosion parameter:

other: other: absorption at 570 nm

Value:

2.22

Remarks on result:

other:

Remarks:

Basis: other: test item. Time point: 60 min. Max. score: 2.27. Reversibility: no data. Remarks: OD mean 2.27 = 100 % viability; OD mean 2.22 = 98 % viability. (migrated information)

In vivo

Irritant / corrosive response data:

Compound Cell viability after 3 min. [%] Cell viability after 60 min. [%] Classification

Diazobarbitursäure 101.14 98.15 Non-Corrosive
Negative control 100.00 100.00 Negative Control

Any other information on results incl. tables

Sample No	Test item	Time [min.]	OD mean*	StdDev	Viability %
1 -3	control NaCl 0.9%	60	2.27	0.06	100
7 -9	Diazobarbitursäure	60	2.22	0.17	98.15
10 -12	control NaCl 0.9%	3	2.27	0.04	100
16 -18	Diazobarbitursäure	3	2.30	0.20	101.14

* 6 values

Applicant's summary and conclusion

Interpretation of results:

other: not corrosive

Executive summary:

An in-vitro study for predicting non-specific, corrosive potential of Diazobarbitursäure by using reconstructed human skin (RHS) EST-1000 was performed in accordance with OECD TG 431 and with EC guideline (amending Concil Direcrive 67/548/EEC B 40 Skin corrosion). Diazobarbitursäure was tested undissolved (25 mg per insert, plus 50 µl 0.9 % NaCl to moisten and to ensure good contact with the skin). An incubation time of 3 min or of 60 min was used to determine time related toxic effects. After incubation the MTT reduction assay was performed to measure the cell viability which was 101 % and 98 % , respectively. Thus, the results show that no corrosive property of the test item was determined by the assay used (Wingenroth 2012).