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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study performed according to GLP and guideline

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2001

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Qualifier:
according to
Guideline:
OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)
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): Calcium sulfate dihydrate
- Analytical purity: 99.9%
- Lot/batch No.: Sigma Aldrich Corporation, Lot No. - 06316BO

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:
Rat (Sprague-Dawley strain), male, liver homogenate - S9 mix
Test concentrations with justification for top dose:
12, 37, 111, 333, 1000 and 3000 µg/plate
Controlsopen allclose all
Untreated negative controls:
yes
Remarks:
0 µg/plate of test material
Negative solvent / vehicle controls:
no
True negative controls:
no
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
Used with tester stratins TA100 and TA1535 at 0.5 µg/plate
Untreated negative controls:
yes
Remarks:
0 µg/plate of test material
Negative solvent / vehicle controls:
no
True negative controls:
no
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
Remarks:
Used with tester strains WP2 uvrA and TA98 at 0.5 µg/plate
Untreated negative controls:
yes
Remarks:
0 µg/plate of test material
Negative solvent / vehicle controls:
no
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
Used with tester starins TA1537 at 50 µg/plate
Untreated negative controls:
yes
Remarks:
0 µg/plate of test material
Negative solvent / vehicle controls:
no
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2 aminoanthracene
Remarks:
Used with tester strains TA100 and TA98 at 0.4 µg/plate and tester strains TA1535 and TA1537 at 2 µg/plate and tester strain WP2 uvrA at 4 µg/plate
Details on test system and experimental conditions:
Description of follow up repeated study: Preliminary test had carried out to decide the appropriate starting dose level of the main study at the concentration of 1.6, 8, 40, 200, 1,000 and 3,000 μg/plate. Criteria for evaluating results: the number of revertant colonies in the plate was counted after 2 days incubation at 37 °C.

The direct incorporation method: For test without metabolic activation, the test substance and 0.1 ml of fresh bacterial culture were added to 2.0 ml of overlay agar. For tests with metabolic activation, 0.5 ml of metabolic activation mixture containing an adequate amount of postmitochondrial fraction was added to the overlay agar after the addition of the bacteria and test substance. All plates in a given test should be incubated for the same time period.





Results and discussion

Test resultsopen allclose all
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
Vehicle controls validity:
not applicable
Untreated negative controls validity:
not applicable
Positive controls validity:
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:
not applicable
Untreated negative controls validity:
valid
Positive controls validity:
not valid

Any other information on results incl. tables

Table 1: Result of bacterial reverse mutation assay with calcium sulfate, dihydrate.

Tester strain

Chemical treated

Dose (µg/plate)

Colonies/plate (mean)[Factor]

Without S-9 mix

With S-9 mix

TA100

Test item

0

12

37

111

333

1,000

3,000

132

139 [1.0]

143 [1.1]

125 [0.9]

129 [1.0]

124 [0.9]

124 [0.9]

114

115 [1.0]

122 [1.1]

115 [1.0]

125 [1.0]

106 [1.0]

115 [1.0]

TA 1535

Test item

0

12

37

111

333

1,000

3,000

13

20 [1.5]

22 [1.7]

17 [1.3]

16 [1.2]

17 [1.3]

14 [1.1]

10

11 [1.1]

12 [1.2]

13 [1.3]

14 [1.4]

13 [1.3]

9 [0.9]

TA 1537

Test item

0

12

37

111

333

1,000

3,000

15

16 [1.0]

11 [0.7]

15 [1.0]

14 [0.9]

13 [0.9]

12 [0.8]

17

17 [1.0]

22 [1.3]

19 [1.1]

17 [1.0]

16 [0.9]

16 [0.9]

E.coli

WP2uvrA

Test item

0

12

37

111

333

1,000

3,000

8

7 [0.9]

6 [0.8]

8 [1.0]

8 [1.0]

6 [0.8]

6 [0.8]

 

12

8 [0.7]

13 [1.1]

8 [0.7]

8 [0.7]

9 [0.8]

9 [0.8]

Positive controls

 

 

 

TA100

TA1535

TA98

TA1537

WP2uvrA

TA100

TA1535

TA98

TA1537

WP2uvrA

SA

SA

4NQO

9-AA

4NQO

2-AA

2-AA

2-AA

2-AA

2-AA

0.5

0.5

0.5

50

0.5

0.4

2

0.4

2

4

493 [3.7]

371 [28.5]

426 [19.4]

740 [49.3]

377 [47.1]

 

 

 

 

14 [0.6]

 

 

 

 

 

491 [4.3]

394 [39.4]

289 [8.5]

326 [19.2]

311 [25.9]

[Factor]: No. of colonies of treated plate/No. of colonies of negative control plate

SA: Sodium azide 9-AA: 9-Amino acridine

4NQQ: 4-nitroquinoline-1-oxide 2-AA: 2-aminoanthracene

Applicant's summary and conclusion

Conclusions:
Interpretation of results: negative With and without metabolic activation

Mutation in the Salmonella tryphimurium (strains TA 98, TA100, TA 1535 and TA 1537) and in the Escherichia coli WP2 uvrA did not occur with calcium sulfate, dihydrate