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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:
2009-02-26 - 20009-04-23
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The GLP study was conducted according to an internationally accepted guideline. All study parameters are given in detail.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2009

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: crystalline
Details on test material:
Name: Ukanol FR 70
Batch no.: 4251318
Appearance: white solid
Composition: 9,10-Dihydro-9-oxa~10-[2,3-Di-(2-hydroxyethoxy) carbonylpropyl]-10-phosphaphenanthren-10-oxid; Bis(2-hydroxyethyl)-(6H-dibenz[c,e][1,2]oxaphos-phorin-6-yl-methyl)succinat-P-oxide
CAS No.: 63562-34-5
EINECS-No.: 264-313-6
Molecular formula: C21H23O8P
Molecular weight: 434 g/mo
Purity: app. 90% (HPLC)
Homogeneity: homogeneous

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 97
Additional strain / cell type characteristics:
other: hisD6610, uvrB, pKM 101, rfa
Species / strain / cell type:
S. typhimurium TA 98
Additional strain / cell type characteristics:
other: hisD3052, uvrB, pKM 101, rfa
Species / strain / cell type:
S. typhimurium TA 100
Additional strain / cell type characteristics:
other: hisG46, uvrB, pKM 101, rfa
Species / strain / cell type:
S. typhimurium TA 102
Additional strain / cell type characteristics:
other: hisG428, pKM 101, rfa
Species / strain / cell type:
S. typhimurium TA 1535
Additional strain / cell type characteristics:
other: hisG46, uvrB, rfa.
Metabolic activation:
with and without
Metabolic activation system:
S9, produced from the livers of male Sprague-Dawley rats which were treated with 500 mg Aroclor 1254/kg body weight intraperitoneally.
Test concentrations with justification for top dose:
First Experiment:
1501 / 500 / 150 / 50 /15 µg/plate

Second Experiment:
1502 / 751 / 376 / 188 / 94 µg/plate

Vehicle / solvent:
DMSO
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
sodium azide
benzo(a)pyrene
other: 4-Nitro-1,2-phenylene diamine, 2-Amino-anthracene,
Details on test system and experimental conditions:
On the day of the start of the first experiment, a stock solution containing 50 g/L of the test item in DMSO was prepared.
In the second experiment, a stock solution containing 15 g/L of the test item in DMSO was prepared.
DMSO was chosen as solvent, because the test item was completely soluble, and this solvent doesn't have any effects on the viability of the bacteria or the number of spontaneous revertants. The stock solutions were used to prepare the geometric series of the concentrations to be tested. Each solution was membrane filtrated to accomplish sterility.
One day before the start of each experiment, one pellet per strain to be used was put into a culture vessel containing nutrient broth. For the incubation of strains TA97a, TA98, TA100 and TA102, ampicilline was added to the medium (25 mg/L). For the incubation of strain TA102, additionally, tetracycline was added (20 mg/L). TA1535 was incubated without addition of antibiotica, as this strain doesn't carry a resistance plasmid.

After incubation for 10 hours at 37 °C, the cultures were used in the experiment. During the test, the cultures were stored at room temperature as to prevent changes in the titre.
Evaluation criteria:
A test item is considered to have mutagenic potential, if a significant, reproducible increase of revertant colonies per plate (increase factor > 2) in at least one strain can be observed. A concentration-related increase over the range tested can also be taken as a sign of mutagenic activity.

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 97
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 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
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
Key result
Species / strain:
S. typhimurium TA 102
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 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

Any other information on results incl. tables

The mean revertant values of the four replicates are presented in the following table.

Strain

TA97a

TA98

TA100

TA102

TA1

535

Induction

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

H20

Mean

n.t.

n.t.

n.t.

n.t.

114

n.t.

n.t.

n.t.

10

n.t.

sd

n.t.

n.t.

n.t.

n.t.

15.3

n.t.

n.t.

n.t.

3.3

n.t.

DMSO

Mean

103

97

10

8

99

131

209

215

9

11

sd

5.8

13.0

5.4

3.9

7.4

17.7

65.4

26.7

3.1

3.3

Pos.Contr.

Mean

1001

1001

1001

1001

1001

1001

1001

1001

1001

1001

sd

0

0

0

0

0

0

0

0

0

0

m

9.72

10.32

100.1

125.1

8.78

7.64

4.79

4.66

100.1

91.00

1502|jg/pl.

Mean

86

82

7

6

105

111

152

181

11

8

sd

5

6

2

1

17

18

10

64

4

2

m

0.83

0.85

0.70

0.75

1.06

0.85

0.73

0.84

1.22

0.73

751Mg/pi.

Mean

109

88

6

6

95

114

132

222

8

14

sd

5

20

1

1

20

6

20

19

1

6

m

1.06

0.91

0.60

0.75

0.96

0.87

0.63

1.03

0.89

1.27

376ng/pi.

Mean

83

87

8

6

120

99

236

201

11

11

sd

8

15

4

1

6

15

72

22

2

1

m

0.81

0.90

0.80

0.75

1.21

0.76

1.13

0.93

1.22

1.00

188Mg/pl-

Mean

123

111

6

6

104

92

134

175

10

12

sd

11

6

2

5

22

15

5

45

3

5

m

1.19

1.14

0.60

0.75

1.05

0.70

0.64

0.81

1.11

1.09

94jig/pi.

Mean

106

100

9

5

150

99

157

213

13

10

sd

8

16

2

3

11

1

13

9

3

2

m

1.03

1.03

0.90

0.63

1.52

0.76

0.75

0.99

1.44

0.91

Applicant's summary and conclusion

Conclusions:
Interpretation of results:
negative

Under the test conditions, the test item 9,10-Dihydroxy-9-oxa-10-[2,3-Di-(2-hydroxyethoxy)carbonylpropyl]-10-phosphaphenanthren-10-oxid is not mutagenic in the Bacterial Reverse Mutation Test using Salmonella typhimurium, strains TA97a, TA98, TA100, TA102 and TA1535.
Executive summary:

In a reverse gene mutation assay in bacteria according to OECD guideline 471, strains TA 1535, TA 97, TA98,TA100 and TA 102 of S. typhimurium were exposed to 9,10-Dihydroxy-9-oxa-10-[2,3-Di-(2-hydroxyethoxy)carbonylpropyl]-10-phosphaphenanthren-10-oxid.

The test item didn't show mutagenic effects in both experiments. The number of revertant colonies was not increased in comparison with the spontaneous revertants (solvent only). Cytotoxicity of the test item was not detected: The background lawn was visible and the number of revertants was not significantly decreased. Therefore it can be stated, that under the test conditions, the test item 9,10-Dihydroxy-9-oxa-10-[2,3-Di-(2-hydroxyethoxy)carbonylpropyl]-10-phosphaphenanthren-10-oxid is not mutagenic in the Bacterial Reverse Mutation Test using Salmonella typhimurium, strains TA97a, TA98, TA100, TA102 and TA1535.