2-[4-[[2-(cyanoimino)hexahydro-4,6-dioxo-5-pyrimidyl]azo]phenyl]-6-methylbenzothiazole-7-sulphonic acid, compound with 2,2',2''-nitrilotris[ethanol] (1:1)

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1990-02-14 to 1990-03-12

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

5 strains tested but not including E. coli

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1990

Report date:

1990

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

1983

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Version / remarks:

1984

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals

Version / remarks:

1984

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2-[4-[[2-(cyanoimino)hexahydro-4,6-dioxo-5-pyrimidyl]azo]phenyl]-6-methylbenzothiazole-7-sulphonic acid, compound with 2,2',2''-nitrilotris[ethanol] (1:1)

EC Number:

259-470-2

EC Name:

2-[4-[[2-(cyanoimino)hexahydro-4,6-dioxo-5-pyrimidyl]azo]phenyl]-6-methylbenzothiazole-7-sulphonic acid, compound with 2,2',2''-nitrilotris[ethanol] (1:1)

Cas Number:

55067-15-7

Molecular formula:

C19 H13 N7 O5 S2 . C6 H15 N O3

IUPAC Name:

2-[4-[[2-(cyanoimino)hexahydro-4,6-dioxo-5-pyrimidyl]azo]phenyl]-6-methylbenzothiazole-7-sulphonic acid, compound with 2,2',2''-nitrilotris[ethanol] (1:1)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Op. 604/89
- Expiration date of the lot/batch: December, 1991

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: in solvent: room temperature, protected from light
- Stability under test conditions: 2 hours stable in water and DMSO

Method

Species / strain

Species / strain / cell type:

other: Salmonella typhimurium strain TA 1535, TA 1537, TA 1538, TA 98, TA 100

Metabolic activation:

with and without

Metabolic activation system:

S9 fraction from rat liver, Aroclor 1254 induced

Test concentrations with justification for top dose:

10.0; 100.0; 333.3; 1000; and 5000 µg/plate

Vehicle / solvent:

- Vehicle/solvent used: deionized water
- Justification for choice of solvent/vehicle: good solubility of the test substance in water

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 4-nitro-o-phenylene-diamine

Controls 3

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 72 hours at 37 °C
- Expression time (cells in growth medium): 72 hours at 37 °C

SELECTION AGENT (mutation assays): histidine

NUMBER OF REPLICATIONS: 3

NUMBER OF CELLS EVALUATED: all revertant cells

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth

Evaluation criteria:

A test article is considered as mutagen if in strain TA 100 the number of reversions is at least twice as high and in strains TA 1535, TA 1537, TA 1538, and TA 98 it is at least three times higher as compared to the spontaneous reversion rate.
Also, a dose-dependent increase in the number of revertants is regarded as an indication of possibly existing mutagenic potential of the test article regardless whether the highest dose induced the above described enhancement factors or not.

Statistics:

No appropriate statistical method is available.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Table 1: Number of revertants per plate (mean of three plates), experiment 1

	strain TA 1535		strain TA 1537		Strain TA 1538		strain TA 98		strain TA 100
conc. [µg]	-MA	+MA	-MA	+MA	-MA	+MA	-MA	+MA	-MA	+MA
Negative control	6	7	5	5	19	18	18	23	73*	82
Solvent control	7	8	4	5	16	19	17	22	79	75
10	8	7	5	5	14	18	19	26	77	68
100	5	5	4	6	15	17	19	26	71	74
333.3	5	8	5	3	18	20	19	21	72	57
1000	6	6	4	6	16	18	17	27	69	53
5000	5	7	2	5	17	20	19	26	73	70
Positive control sodium azide (10 µg)	738		146		1475		760		656
Positive control 2-Aminoanthracene (10 µg)		200		37		78		1440		1236

*Unexpected low revertant rate

  

Table 2: Number of revertants per plate (mean of three plates), experiment 2

	strain TA 1535		strain TA 1537		Strain TA 1538		strain TA 98		strain TA 100
conc. [µg]	-MA	+MA	-MA	+MA	-MA	+MA	-MA	+MA	-MA	+MA
Negative control	21	16	6	8	15	17	15	30	75	99
Solvent control	21	17	6	9	21	23	15	27	78	88
10	29	21	7	6	22	16	18	27	71	79
100	19	22	7	9	21	30	14	24	66	84
333.3	16	21	4	10	21	30	13	31	73	88
1000	18	20	5	10	22	32	15	32	67	94
5000	17	16	6	8	24	22	16	43	71	80
Positive control sodium azide (10 µg)	1335		217		1345		1522		1291
Positive control 2-Aminoanthracene (10 µg)		65		107		547		1667		573