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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:
guideline study
Remarks:
Guideline study, GLP study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1998
Report date:
1998

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
(only 2-aminoanthracene as positive control with metabolic activation)
Qualifier:
according to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
2,2'-dimorpholinyldiethyl ether
EC Number:
229-194-7
EC Name:
2,2'-dimorpholinyldiethyl ether
Cas Number:
6425-39-4
Molecular formula:
C12H24N2O3
IUPAC Name:
4-{2-[2-(morpholin-4-yl)ethoxy]ethyl}morpholine
Test material form:
liquid
Details on test material:
- State of aggregation : liquid
- Appearance : colourless to yellow brown
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: H 50541 H x B (Substance number: 95/452)
- Analytical purity: 98.4%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature (N2 conditions)
- Stability under test conditions: guaranteed throughout study period via reanalysis

OTHER SPECIFICS:
- Name of test substance (as cited in study report): 2,2' -Dimorpholinodiethylether
- Physical state: liquid

Method

Target gene:
His- and Trp-Operon
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:
Aroclor 1254-induced rat liver S9-mix
Test concentrations with justification for top dose:
0, 20, 100, 500, 2,500, 5,000 µg/plate (standard plate test and preincubation test)
Vehicle / solvent:
water
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
other: with S9: 2-aminoanthracene (all tester strains); without S9: N-methyl-N'-nitro-N-nitrosoguanidine (TA 1535, TA 100), 4-nitro-o-phenylendiamine (TA 98), 9-aminoacridine (TA 1537), N-ethyl-N'-nitro-N-nitrosoguanidine (E. coli WP2 uvrA)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation); preincubation

DURATION (preincubation test)
- Preincubation period: 20 min
- Exposure duration: 48 - 72 h

DURATION (standard plate test)
- Exposure duration: 48 - 72 h

NUMBER OF REPLICATIONS: 3 plates/dose/experiment.


DETERMINATION OF CYTOTOXICITY
- Method: reduction of the number of revertants compared to negative control; reduction of titer; reduction of backgroud growth

Evaluation criteria:
The test subtance is considered positive in this assay if the following criteria are met: A dose-related and reproducible increase in the number of revertant colonies, i.e. about doubling of the spontaneous mutation rate in at least one tester strain either without S9-mix or after adding a metabolizing system.

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:
valid
Untreated negative controls validity:
valid
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:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: no precipitation up to 5,000 µg/plate (highest dose tested)
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Standard plate test:

 Dose (µg/plate)

 TA1535   

 TA100   

 TA1537   

 TA98   

    E. coli WP2 uvrA

 

 -S9

 +S9

 -S9

 +S9

 -S9

 +S9

-S9

 +S9

-S9 

+S9 

 0

20±5

19±2

128±10

123±4

10±1

10±1

31±4

47±5

32±1

42±2

20 

19±2

18±2

115±20

125±12

10±2

11±4

32±3

46±4

33±5

41±3

100

19±5

17±3

111±7

121±14

10±1

7±1

31±5

39±8

30±2

37±6

 500

19±3

14±1

128±8

107±14

9±3

8±1

27±5

33±10

30±1

39±2

 2500

19±3

16±4

123±7

121±15

10±1

6±0

31±4

38±5

32±4

42±2

 5000

18±3

15±2

140±3

97±17

8±1

8±1

28±6

28±8

30±3

43±2

 2 -AA

-

149±7

-

1467±

133

 -

149±24

1241±

29

 -

236±

17

 MNNG

783±

37

-

1027±

49

-

-

 -

-

 -

-

-

 AAC

-

-

-

-

804±13

-

-

-

-

-

 NOPD

 -

-

-

-

-

-

1308±

135

-

 ENNG

 -

-

-

-

-

-

-

-

521±

26

-

Mean ± SD

 

Preincubation-test:

 Dose (µg/plate)

 TA1535   

 TA100   

 TA1537   

 TA98   

 E. coli WP2

uvrA   

 

 -S9

 +S9

 -S9

 +S9

 -S9

 +S9

-S9

 +S9

 -S9

+S9 

 0

18±1

19±2

133±4

130±17

11±2

11±3

29±1

40±3

32±4

38±3

20

18±2

16±2

116±9

108±9

11±1

11±2

23±2

42±1

31±6

32±5

 100

21±1

17±0

124±6

121±10

8±1

11±1

24±2

36±5

28±4

33±5

 500

17±2

14±1

111±0

125±13

8±2

11±2

24±2

38±1

30±6

25±4

 2500

19±1

16±3

117±15

132±7

8±2

9±1

25±1

40±3

28±3

28±4

 5000

16±2

15±2

102±16

138±14

7±1

7±2

20±3

38±7

26±2

30±51

2-AA

-

116±13

-

699±

2

-

119±9

-

727±

12

-

177±11

 MNNG

978±

135

-

1177± 126

-

-

-

-

-

-

-

 AAC

-

-

-

-

402±62

-

-

-

-

-

NOPD

-

-

-

-

-

-

1148±

103

-

-

-

 ENNG

-

 -

 -

 -

-

 -

-

569±49

 -

Mean ± SD

X: reduced background growth

2-AA: 2-aminoanthracene

MNNG; N-methyl-N-nitro-N-nitrosoguanidine

ENNG; N-ethyl-N-nitro-N-nitrosoguanidine

NOPD: 4-nitro-o-phenylendiamine

AAC: 9-aminoacridine chloride monohydrate

 

According to the results of the study, the test substance is not mutagenic in the Ames test and in the Escherichia coli - reverse mutation assay under the experimental conditions chosen here. The positive controls gave the expected results.

 

Applicant's summary and conclusion

Conclusions:
According to the results of the study, the test substance is not mutagenic in the Ames test and in the Escherichia coli - reverse mutation assay under the experimental conditions chosen here. The positive controls gave the expected results.