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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: GLP-Guideline study.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1997
Report Date:
1997

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
; OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)
Deviations:
yes
Remarks:
(2-aminoanthracene was uased as sole indicator of the efficacy of S9-mix)
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): N-Butylethanolamin
- Physical state: colorless to yellowish liquid
- Analytical purity: 99.8%
- Batch No.: 89-9620
- Storage condition of test material: room temperature

Method

Target gene:
His- and Trp-Operon
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Metabolic activation:
with and without
Metabolic activation system:
Aroclor-induced rat liver S9-mix
Test concentrations with justification for top dose:
20, 100, 500, 2500, 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: water
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
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

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 - 72 h at 37°C

NUMBER OF REPLICATIONS: 3

METHOD OF APPLICATION: standard plate test

DURATION
- Exposure duration: 48 - 72 h at 37°C

NUMBER OF REPLICATIONS: 3
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.
A test substance is generally considered nonmutagenic in this test if: The number of revertants for all tester strains were within the historical negative control range under all experimental conditions in two experiments carried out independently of each other.

Results and discussion

Test results
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
other: A bacteriotoxic effect was observed in the plate incorporation test depending on the test conditions from about 2500 µg/plate onward.
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
SOLUBILITY: No precipitation of the test substance was found.
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±1

21±2

136±4

153±7

14±1

13±2

37±4

46±1

36±1

41±4

 20

27±1

21±1

135±18

169±12

12±3

11±2

25±1

42±2

32±1

42±3

 100

22±1

20±3

147±22

136±15

10±2

14±2

23±1

44±4

33±3

45±3

 500

27±4

19±4

168±15

183±11

18±1

14±2

26±6

41±3

35±3

37±1

 2500

28±3

23±6

169±13

189±4

12±1

13±1

28±1

36±6

33±7

42±5

 5000

25±6

26±6

165±9

162±10

13±3

13±4

24±3

21±2

26±6

40±1

 2-AA

-

219±16

-

1276±53

-

220±21

-

1284±216

-

352±67

 MNNG

1093±35

-

1063±59

-

-

-

-

-

-

-

 AAC

-

-

-

-

648±53

-

-

-

-

-

 NPD

-

-

-

-

-

-

809±76

-

-

-

 ENNG

-

-

-

-

-

-

-

-

668±30

-

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

17±1

15±1

106±11

109±9

10±1

10±2

30±2

41±3

31±2

38±3

 20

17±2

18±2

109±4

111±20

10±2

9±1

28±5

36±5

32±6

33±3

 100

17±1

17±1

115±2

102±11

9±1

9±2

28±1

40±2

30±3

39±7

 500

17±3

19±2

106±5

97±4

6±1

8±1

28±3

45±3

31±2

35±3

 2500

7±2

17±1

15±3

97±17

4±2

7±1

4±5

40±3

21±4

26±5

 5000

-

9±1

-

68±13

-

5±2

-

28±3

-

14±2

 2-AA

-

229±8

-

753±39

-

136±8

-

919±79

-

211±6

 MNNG

1067±61

-

1137±21

-

-

-

-

-

-

-

 AAC

-

-

-

-

694±27

-

-

-

-

-

 NPD

-

-

-

-

-

-

1013±25

-

-

-

 ENNG

-

-

-

-

-

-

-

-

504±5

-

Mean ± SD

2-AA: 2-aminoanthracene (2.5 µg/p1ate, strains: TA 1535, TA 100, TA 1537, TA 98; 60 µg/p1ate, strain: Escherichia coli WP2 uvrA)

MNNG; N-methyl-N-nitro-N-nitrosoguanidine (5 µg/p1ate, strains: TA 1535, TA 100)

ENNG; N-ethyl-N-nitro-N-nitrosoguanidine (10 µg/p1ate, strain: E. coli WP2 uvrA)

NPD: 4-nitro-o-phenylendiamine (10 µg/p1ate, strain: TA 98)

AAC: 9-aminoacridine chloride monohydrate (100 µg/plate, strain: TA 1537)

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

Under the conditions tested butylethanolamine is non mutagenic in the bacterial AMES-test with or without metabolic activation.
Executive summary:

In a reverse gene mutation assay in bacteria (BASF, 1997), strains [TA 1535, TA 1537, TA 98, TA 100] of S. typhimurium and E. coli [E. coli WP2] were exposed to the test substance dissolved in water at concentrations of 20, 100, 500, 2500, 5000 µg/plate in the absence of mammalian metabolic activation and in the presence of Aroclor 1254-induced rat liver S9 mix, respectively (the preincubation method and the standard plate test was used). A bacteriotoxic effect was observed in the plate incorporation test depending on the test conditions from about 2500 µg/plate onward.No significant increase in mutant frequency was observed, either with or without metabolic activation. The positive controls induced the appropriate responses in the corresponding strains.