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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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: OECD guideline study (OECD 471/472) conducted in compliance with GLP regulations

Data source

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

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
and OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay) (May 1983/Revised Draft, March 1996)
Deviations:
yes
Remarks:
Compared to OECD TG 471 (1997), only one positive control substance (2AA) used as the sole indicator of the efficacy of the S9-mix, a second control substance is missing.
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): Dimethylaminodiglycol
- Physical state: Colorless to yellowish liquid
- Analytical purity: 99.1 area-% (GC)
- Storage condition of test material: Room temperature

Method

Target gene:
His +/-; Trp +/-
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:
The S9 fractions were prepared from the liver of Aroclor 1254-induced male Sprague-Dawley rats.
Test concentrations with justification for top dose:
0; 20; 100; 500; 2500 and 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: water
- Justification for choice of solvent/vehicle: good solubility
Controls
Untreated negative controls:
yes
Remarks:
sterility control
Negative solvent / vehicle controls:
yes
Remarks:
water control
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: With S-9 mix: all strains 2-aminoanthracene; without S-9 mix: strains TA100, TA1535: N-methyl-N'-nitro-N-nitrosoguanidine; TA98: 4-nitro-o-phenylendiamine; TA1537: 9-aminoacridine; E.coli WP2 uvrA: N-ethyl-N'-nitro-N-nitrosoguanidine.
Details on test system and experimental conditions:
METHOD OF APPLICATION: preincubation test

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 3


METHOD OF APPLICATION: standard plate test (plate incorporation assay)

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 3


DETERMINATION OF CYTOTOXICITY (in both assays)
- Method: Toxicity was detected by a
- decrease in the number of revertants
- clearing or diminution of the background lawn (= reduced his- or trp- background growth)
- reduction in the titer.
Evaluation criteria:
The test chemical 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 S-9 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:
cytotoxicity
Remarks:
A slight decrease in the number of revertants was occasionally observed depending on the strain and test conditions at doses > 2500 µg/plate.
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:
cytotoxicity
Remarks:
A slight decrease in the number of revertants was occasionally observed at doses of 5000 µg/plate.
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: No test substance precipitation was found.

Any other information on results incl. tables

Standard plate test (mean ± SD):

 Dose (µg/plate)  TA1535     TA100     TA1537     TA98        E. coli WP2 uvrA                                                                        
   -S9  +S9  -S9  +S9  -S9  +S9 -S9  +S9 -S9  +S9                                                                         
 0

21±2

23±1

117±6

117±3

13±1

16±2

29±2

45±4

32±2

47±3

 

 

 

 

 

 

                                                           

20 

20±1

20±1

113±7

109±7

13±1

12±1

26±2

35±1

32±4

42±4

 

 

 

 

 

 

                                                           

 100

17±2

18±3

107±11

107±3

13±3

11±1

24±3

33±1

32±5

39±2

 

 

 

 

 

 

                                                           

 500

19±3

20±1

109±15

115±19

11±2

9±2

23±2

31±3

34±4

34±1

 

 

 

 

 

 

                                                           

 2500

19±2

19±1

104±10

123±9

11±2

8±1

24±3

28±1

31±3

33±3

 

 

 

 

 

 

                                                           

 5000

6±1

17±3

100±8

115±13

12±1

11±1

22±2

29±2

28±2

36±5

 

 

 

 

 

 

                                                           

 2 -AA

-

149±25

-

1243±212

 -

110±5

1127±86

 -

206±23

 

 

 

 

 

 

                                                           

 MNNG

1048±64

-

1116±75

-

-

 -

-

 -

-

-

 

 

 

 

 

 

                                                           

 AAC

-

-

-

-

732±64

-

-

-

-

-

 

 

 

 

 

 

                                                           

 NOPD

 -

-

-

-

-

-

728±62

-

 

 

 

 

 

 

                                                           

 ENNG

 -

-

-

-

-

-

-

-

749±69

-

 

 

 

 

 

 

                                                           

Preincubation-test (mean ± SD):

 Dose (µg/plate)

 TA1535   

 TA100   

 TA1537   

 TA98   

 E. coli WP2

uvrA   

 

 

 

 

 

 

 

                                                       

 

 -S9

 +S9

 -S9

 +S9  -S9  +S9 -S9  +S9  -S9 +S9                                                                       
 0

21±2

20±2

129±9

150±17

12±1

13±2

31±1

46±6

33±2

49±5

 

 

 

 

 

 

 

                                                       

20

20±3

19±1

121±8

166±12

10±1

10±1

31±1

43±2

32±3

45±5

 

 

 

 

 

 

 

                                                       

 100

18±3

18±2

123±11

162±11

10±1

11±1

31±3

42±4

31±4

40±2

 

 

 

 

 

 

 

                                                       

 500

15±2

18±1

117±17

159±13

9±1

8±1

26±3

41±3

26±4

41±3

 

 

 

 

 

 

 

                                                       

 2500

13±1

16±3

145±16

160±13

9±2

7±2

23±3

31±1

26±7

37±2

 

 

 

 

 

 

 

                                                       

 5000

9±2

15±2

126±7

163±15

6±1

8±1

24±2

31±1

22±2

31±1

 

 

 

 

 

 

 

                                                       

2-AA

-

170±29

-

1593±105

-

135±32

-

1380±205

-

233±

13

 

 

 

 

 

 

 

                                                       

 MNNG

2029±73

-

2281±169

-

-

-

-

-

-

-

 

 

 

 

 

 

 

                                                       

 AAC

-

-

-

-

644±58

-

-

-

-

-

 

 

 

 

 

 

 

                                                       

NOPD

-

-

-

-

-

-

871±140

-

-

-

 

 

 

 

 

 

 

                                                       
 ENNG -  -  -

 - -  - - 764±5  -                                                                      

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

Under the conditions tested 2-[2-(dimethylamino)ethoxy]ethanol is non mutagenic in the bacterial AMES-test.

The positive control gave the expected response.

Applicant's summary and conclusion