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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
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study with restrictions. (Only 4 test strains used and only 2-Aminoanthracene is used as positive control for S9 mix. No testing for cross-linking using TA102 or WP strains.)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1992
Report date:
1992

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
adopted 21 July 1997
Deviations:
yes
Remarks:
only 4 tester strains used; no testing for crosslinking using TA102 or E.coli WP2 strains; 2-AA sole positive control used for S9 mix
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
2-[2-(3-aminopropoxy)ethoxy]ethanol
EC Number:
203-960-0
EC Name:
2-[2-(3-aminopropoxy)ethoxy]ethanol
Cas Number:
112-33-4
Molecular formula:
C7H17NO3
IUPAC Name:
2-[2-(3-aminopropoxy)ethoxy]ethan-1-ol
Details on test material:
- Name of test material (as cited in study report): 9-Amino-3,6-dioxanonan-1-ol
- Physical state: colorless liquid
- Analytical purity: 99.23 %
- Lot/batch No.: 1/9/3
- Test substance No.: 91/455
- Storage condition of test material: room temperature

Method

Target gene:
his operon
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
other: uvrB (all strains), rfa (all strains), R factor plasmid (TA98 + TA100)
Metabolic activation:
with and without
Metabolic activation system:
co-factor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats pretreated for 5 days with 500 mg/kg bw Aroclor 1254
Test concentrations with justification for top dose:
first (standard plate test) and second (preincubation test) experiment:
20, 100, 500, 2500, 5000 µg/plate
with and without metabolic activation
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: water
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
water
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-Aminoanthracene (2-AA, 2.5µg/plate; all +S9); N-methyl-N'-nitro-N-N-nitroso-guanidine (MNNG, 5µg/plate, TA100 & TA 1535, -S9), 4-nitro-o-phenylendiamine chloride monohydrate (NPD, 10µg/plate; TA98 -S9); 9-Aminoacridine (AAC, 100µg/plate; TA1537 -S9)
Remarks:
all positive controls dissolved in DMSO
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium; with and without preincubation

DURATION
- Preincubation period: 20 minutes
- Exposure duration: 48 hours

NUMBER OF REPLICATIONS: 3 replications in 1 experiment each for standard plate test and preincubation test

DETERMINATION OF CYTOTOXICITY
- Method: reduced background growth and decrease in number of his+ revertants
Evaluation criteria:
In general, a substance to be characterized as positive in the AMES Test has to fulfill the following requirements:
- doubling of the spontaneous mutation rate (control)
- dose-response relationship
- reproducibility of the results
Statistics:
mean values and standard deviations were calculated

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:
cytotoxicity
Remarks:
in all tester strains depending on strain and test condtions at >2500 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
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:
cytotoxicity
Remarks:
only with TA1537 and TA98 at 5000µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: none observed
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'. Remarks: preincubation test

Any other information on results incl. tables

table1: Test results of experiment I (standard plate test)

With or without S9 mix

Test substance concentration [µg/plate]

Mean number of revertant colonies per plate (average of 3 plates ± standard deviation)

Basepair substitution type

Frameshift type

TA100

TA1535

TA98

TA1537

-

0

84 ± 1

12 ± 3

21 ± 3

8 ± 1

-

20

103 ± 10

12 ± 3

21 ± 3

7 ± 1

-

100

95 ± 5

13 ± 3

14 ± 2

9 ± 2

-

500

68 ± 10

12 ± 4

15 ± 3

4 ± 1

-

2500

95 ± 11

11 ± 2

18 ± 5

6 ± 2

-

5000

74 ± 5

13 ± 2

14 ± 3

5 ± 4

Positive controls,

-S9

Name

 

Conc. [µg/plate]

Mean no. of col./plate (average of 3 ± SD)

MNNG*

 

5

 

880 ± 36

MNNG*

 

5

 

740 ± 31

NPD*

 

10

 

777 ± 50

AAC*

 

100

 

602 ± 58

 

 

 

TA100

TA1535

TA98

TA1537

+

0

91 ± 9

13 ± 2

24 ± 4

9 ± 3

+

20

92 ± 21

11 ± 3

24 ± 3

7 ± 2

+

100

108 ± 10

14 ±3

23 ± 8

7 ± 2

+

500

109 ± 8

10 ± 2

37 ± 8

5 ± 1

+

2500

113 ± 17

13 ± 4

25 ± 4

4 ± 0

+

5000

89 ± 8

12 ± 2

11 ± 2

5 ± 1

Positive controls, +S9

Name

 

Conc. [µg/plate]

Mean no. of col./plate (average of 3 ± SD)

2-AA*

 

2.5

 

1013 ± 21

2-AA*

 

2.5

 

93 ± 15

2-AA*

 

2.5

 

835 ± 42

2-AA*

 

2.5

 

82 ± 7

table2: Test results of experiment II (preincubation test)

With or without S9 mix

Test substance concentration [µg/plate]

Mean number of revertant colonies per plate (average of 3 plates ± standard deviation)

Basepair substitution type

Frameshift type

TA100

TA1535

TA98

TA1537

-

0

100 ± 14

16 ± 4

28 ± 5

8 ± 1

-

20

115 ± 11

15 ± 4

18 ± 4

7 ± 0

-

100

122 ± 10

12 ± 3

23 ± 8

6 ± 2

-

500

126 ± 2

17 ± 3

21 ± 7

8 ± 2

-

2500

87 ± 15

8 ± 1

19 ± 1

3 ± 1

-

5000

58 ± 17

5 ± 4

- ± -

- ± -

Positive controls,

-S9

Name

 

Conc. [µg/plate]

Mean no. of col./plate (average of 3 ± SD)

MNNG*

 

5

 

578 ± 66

MNNG*

 

5

 

571 ± 30

NPD*

 

10

 

744 ± 38

AAC*

 

100

 

412 ± 14

 

 

 

TA100

TA1535

TA98

TA1537

+

0

107 ± 5

13 ± 3

27 ± 7

12 ± 2

+

20

118 ± 10

14 ± 2

24 ± 4

7 ± 1

+

100

125 ± 16

13 ± 1

27 ± 8

9 ± 3

+

500

110 ± 11

11 ± 2

27 ± 3

10 ± 0

+

2500

105 ± 20

12 ± 3

24 ± 6

6 ± 2

+

5000

104 ± 4

13 ± 3

18 ± 3

5 ± 2

Positive controls, +S9

Name

 

Conc. [µg/plate]

Mean no. of col./plate (average of 3 ± SD)

2-AA*

 

2.5

 

561 ± 9

2-AA*

 

2.5

 

82 ± 3

2-AA*

 

2.5

 

556 ± 39

2-AA*

 

2.5

 

78 ± 9

* MNNG - N-methyl-N'-nitro-N-nitroso-guanidine; NPD - 4 -nitro-o-phenylendiamine; AAC - 9 -Aminoacridine chloride monohydrate; 2AA - 2 -Aminoanthracene

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative