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Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2004-08-24 - 2004-10-25
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Cross-reference
Reason / purpose for cross-reference:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2004
Report date:
2004

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
OECD 471 (1997)
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
-
EC Number:
477-690-9
EC Name:
-
Cas Number:
874819-71-3
Molecular formula:
Hill formula: C6H9N4O3P CAS formula: C6H9N4O3P
IUPAC Name:
N-(diaminophosphoryl)-2-nitroaniline
Test material form:
solid: crystalline

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
S9 liver microsomal fraction
Test concentrations with justification for top dose:
Concentration range in the main test (with metabolic activation): 50 ... 5000 µg/plate
Concentration range in the main test (without metabolic activation): 50 ... 5000 µg/plate
Vehicle / solvent:
Solvent: dimethyl sulfoxide
Controlsopen allclose all
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
yes
Positive controls:
yes
Positive control substance:
sodium azide
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
yes
Positive controls:
yes
Positive control substance:
9-aminoacridine
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
yes
Positive controls:
yes
Positive control substance:
2-nitrofluorene
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
yes
Positive controls:
yes
Positive control substance:
methylmethanesulfonate

Results and discussion

Test resultsopen allclose all
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
Key result
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
without
Genotoxicity:
positive
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
positive
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
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:
not valid
Positive controls validity:
not valid
Species / strain:
E. coli WP2 uvr A
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
Additional information on results:
Observations:
An increase in revertant numbers was observed at concentrations from 5000 to 500 µg/plate of the test item
in the bacterial strain Salmonella typhimurium TA 1537
in the experiments without metabolic activation as well
as in the plate incorporation test and in the preincubation
test and in the bacterial strain Salmonella typhimurium
TA 98 in all test variants.
This increase was confirmed by a repetition of all positive
experiments.
Remarks on result:
other: all strains/cell types tested

Any other information on results incl. tables

Bacteria

tester

strain

MA

Exp.

Mean numb

ers of revertants per plate

Evalua­tion

Controls

µg test item per plate

UTC

VC

PC

5000

1000

500

100

50

Salmonella typhimurium

TA1535

no

1St

17.3

16.0

1616.0

14.3

15.7

14.3

17.7

19.3

-

no

2nd

8.0

7.3

710.0

8.0

10.7

11.3

11.0

8.0

-

yes

1st

21.0

13.0

162.7

15.0

17.0

16.3

12.3

15.0

-

yes

2nd

11.3

11.7

156.3

8.0

11.0

13.7

9.0

10.0

-

TA1537

no

1st

7.7

7.3

1973.3

15.7

9.7

13.0

10.7

6.0

 

+

no

1str

16.3

20.7

2368.0

34.0

18.3

n.e.

n.e.

n.e.

+

no

2nd

12.3

11.3

2789.3

72.3

23.7

21.0

17.0

12.7

+

no

2r

11.3

13.7

1170.7

72.3

26.3

n.e.

n.e.

n.e.

+

yes

1st

14.7

9.0

478.0

16.3

9.7

10.3

11.7

11.7

-

yes

2nd

10.3

9.7

384.0

14.3

7.7

12.7

7.0

6.3

-

TA98

no

2St

16.7

19.7

473.7

80.7

40.3

27.0

20.3

20.0

+

no

1st1r

18.3

20.3

566.7

71.0

36.3

26.3

n.e.

n.e.

+

no

2"d

21.7

20.3

476.0

176.0

55.0

33.0

27.3

31.3

+

no

2ndr

17.3

9.7

504.0

166.0

53.7

19.3

n.e.

n.e.

+

vcs

1st

27.7

27.0

2245.3

66.7

38.7

26.7

29.7

22.0

t-

ves

1str

31.7

23.7

2245.3

75.7

28.7

27.3

32.7

n.e.

+

yes

2nd

12.7

11.3

2688.0

29.0

18.0

13.0

12.3

11.3

+

yes

2ndr

33.0

29.0

2634.7

52.3

41.7

29.0

22.7

n.e.

+

TA100

no

1st

192.0

192.3

1701.3

191.3

174.3

192.7

188.0

183.3

-

no

2nd

129.3

111.0

1552.0

152.7

111.3

125.3

120.0

128.3

-

yes

1st

168.7

149.3

1365.3

188.0

164.7

184.7

158.3

150.0

-

yes

2nd

130.3

122.3

2613.3

136.7

122.7

133.3

120.0

116.3

-

Escherichia coli

WP2

uvrA

no

1st

29.0

21.0

304.3

17.3

19.7

27.7

29.7

26.3

-

no

2nd

30.0

33.3

521.3

21.7

26.0

26.3

24.7

35.0

-

yes

1st

32.3

35.3

131.3

28.3

30.0

28.3

31.3

28.0

-

yes

2nd

28.0

32.7

155.3

30.5

31.3

31.7

40.7

36.0

-

Applicant's summary and conclusion

Conclusions:
Interpretation of results :
positive

N-(2-Nitrophenyl)phosphoric triamide induces gene mutation in Salmonella Typhimurium TA 1537 and TA 98 under the experimental conditions described.
Executive summary:

N-(2-Nitrophenyl)phosphoric triamide was tested for a possible potential to induce gene mutation in bacteria according to OECD guideline 471.

As test organisms theSalmonella typhimuriumstrains TA 98, TA 100, TA 1535, TA 1537 and theEscherichia colistrain WP2uvrA were used. The bacteria were exposed toN-(2-Nitrophenyl)phosphoric triamideboth, in the presence and absence of rat liver S9 metabolic activation. Two independent experiments were performed according to the standard plate incorporation method (experiment I) or the pre-incubation method (experiment II), respectively.

The concentrations 5000, 1000, 500, 100 and 50 µg/plate were chosen for the experiments according to the standard plate incorporation method and for the experiments according to the pre-incubation method.

Not any cytotoxicity or precipitation was observed in all experiments.

An untreated, a vehicle control and appropriate positive controls were included into the experimental design. Triplicate plates were scored for each experimental point.

The revertant frequencies of the negative controls were within the expected range and the positive control chemicals induced marked increases in revertant colonies.

N-(2-Nitrophenyl)phosphoric triamideinduced at some experimental points (from 5000 to 500 µg/plate) a reproducible, dose dependent, statistically significant increase of revertant numbers for the bacterial strainSalmonella typhimuriumTA 1537 in the experiments without metabolic activation as well as in the plate incorporation test and in the pre-incubation test and for the bacterial strainSalmonella typhimuriumTA 98 in the experiments with and without metabolic activation as well as in the plate incorporation test and in the pre-incubation test.

Based on the results of the reported study it is concluded that N-(2-Nitrophenyl)phosphoric

triamide induces gene mutation inSalmonella typhimurium under the experimental conditions described.