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Toxicological information

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:
12 May 1995 to 22 May 1995
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Only 4 s. typhimur. strains used with no E coli strain (as per the requirements of current guidance)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1995
Report Date:
1995

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
(1983)
Deviations:
no
Qualifier:
according to
Guideline:
other: EU Method B.14 (Salmonella typhimurium, reverse mutation test) (1992)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OTS 798.5265 (The Salmonella typhimurium Bacterial Reverse Mutation Test)
Version / remarks:
(1984)
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): MKH 3586
- Physical state: powder
- Analytical purity: 98.4%
- Purity test date: 30 Nov 1994
- Lot/batch No.: 17004/93
- Expiration date of the lot/batch: 29 May 1995

Method

Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254
Test concentrations with justification for top dose:
Plate incorporation and preincubation +/-S9: 16, 50, 158, 500, 1581, 5000 ug/plate
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
no
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
10 ug/plate (TA 1535) -S9
Positive controls:
yes
Positive control substance:
other: Nitrofurantoin
Remarks:
0.2 ug/plate (TA100) -S9
Positive controls:
yes
Positive control substance:
other: 4-nitro-1,2-phenylene diamine
Remarks:
10 ug/plate (TA1537, TA98) -S9
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene
Remarks:
3 ug/plate (all strains) +S9

Results and discussion

Test results
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

INITIAL TOXICITY-MUTATION ASSAY:

Six doses of the test material ranging from 16 to 5000 ug/plate ±S9 were evaluated in the plate incorporation test. No precipitation or toxicity was observed up to the limit dose, 5000 ug/plate.

 

CONFIRMATORY MUTAGENICITY ASSAY:

Again no precipitation or cytotoxic effects were observed in any of the strains tested.

 

Table 7.6.1 -1: Bacterial mutation assay, summary of results– Initial (plate incorporation) test

Dose

(ug/plate)

0

16

50

158

500

1581

5000

Positive control

REVERTANTS/PLATE

TA1535 –S9

12 ± 3

14 ± 2

11 ± 3

13 ± 3

10 ± 2

8 ± 3

13 ± 3

782 ± 20

TA1535 +S9

10 ± 1

11 ± 3

12 ± 3

12 ± 3

12 ± 4

12 ± 3

14 ± 3

166 ± 6

TA100 –S9

129 ± 8

95 ± 8

100 ± 13

109 ± 9

97 ± 17

102 ± 5

100 ± 12

264 ± 14

TA100 +S9

103 ± 4

107 ±10

115 ± 14

115 ± 3

109 ± 13

106 ± 7

113 ± 8

1661 ± 126

TA1537 –S9

9 ± 2

10 ± 2

9 ± 3

9 ± 3

8 ± 3

9 ± 2

9 ± 3

119 ± 6

TA1537 +S9

11 ± 2

12 ± 2

8 ± 2

12 ± 3

9 ± 3

8 ± 3

7 ± 1

304 ± 47

TA98 –S9

25 ± 7

22 ± 2

26 ± 3

23 ± 5

20 ± 8

20 ± 5

19 ± 4

176 ± 29

TA98 +S9

26 ± 4

29 ± 5

36 ± 5

28 ± 8

34 ± 4

28 ± 6

28 ± 1

1646 ± 102

 

Table 7.6.1 -2: Bacterial mutation assay, summary of results– Initial (pre-incubation) test

Dose

(ug/plate)

0

16

50

158

500

1581

5000

Positive control

REVERTANTS/PLATE

TA1535 –S9

11 ± 1

14 3

14 ± 4

13 ± 1

12 ± 3

14 ± 3

12 ± 2

717 ± 12

TA1535 +S9

12 ± 3

13 ± 3

15 ± 4

12± 4

14 ± 3

11 ± 3

12 ± 4

218 ± 19

TA100 –S9

126 ± 17

116 ± 8

116 ± 19

114 ± 13

122 ± 3

114 ± 14

89 ± 9

356 ± 17

TA100 +S9

124 ± 3

124 ± 11

113 ± 7

107 ± 17

117 ± 15

118 ± 9

121 ± 13

1562 ± 68

TA1537 –S9

11 ± 1

11 ± 2

12 ± 1

9 ± 3

12± 3

101 ± 4

7 ± 2

124 ± 8

TA1537 +S9

11 ± 2

11 ± 3

9 ± 1

9 ± 2

11 ± 2

10 ± 2

13 ± 1

227 ± 31

TA98 –S9

25 ± 5

23 ± 4

22 ± 3

20 ±4

21 ± 8

26 ±8

26 ± 3

166 ± 24

TA98 +S9

27 ± 3

33 ± 3

32 ± 3

30 ± 6

32 ± 5

30 ± 6

31 ± 4

1216 ± 50

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative with metabolic activation
negative without metabolic activation

Based on the results from this study, MKH 3586 was not mutagenic in the bacterial strains tested, either in the presence or absence of metabolic activation when tested up 5000 ug/plate (the maximum dose in accordance with regulatory guidelines).
Executive summary:

In a reverse gene mutation assay in bacteria,Salmonella typhimuriumstrains TA98, TA100, TA1535, and TA1537 were exposed to MKH 3586 formulated in DMSO. The assay was performed in two phases, using the plate incorporation snd the pre-incubation method.

 

In an initial test, the plate incorporation method was used with dose levels of 16 to 5000 ug/plate in the presence and absence of S9 activation was assessed. No precipitation, cytotoxicity or increase in revertants were observed in any strain tested. The same effects were observed in the pre-incubation, confirmatory test where the same dose range was tested.

 

Based on the results from this study, MKH 3586 was not mutagenic in the bacterial strains tested, either in the presence or absence of metabolic activation when tested up 5000 ug/plate (the maximum dose in accordance with regulatory guidelines).