1,1'-[methylenebis(oxy)]dibutane

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

other information

Study period:

26. May 2000 - 16. Jun 2000

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study (OECD 471)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2000

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

1,1'-[methylenebis(oxy)]dibutane

EC Number:

219-909-0

EC Name:

1,1'-[methylenebis(oxy)]dibutane

Cas Number:

2568-90-3

Molecular formula:

C9H20O2

IUPAC Name:

1,1'-[methylenebis(oxy)]dibutane

Details on test material:

- Name of test material (as cited in study report): Di-n-butylformal
- Physical state: colorless liquid
- Analytical purity: 99.8 %
- Lot/batch No.: Batch 905
- Storage condition of test material: Room temperature (N2 conditions)

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

S9-mix from rat livers (treated with 500 mg Aroclor 1254)

Test concentrations with justification for top dose:

Standarp plate test: 20, 100, 500, 2500, 5000 µg/plate
Preincubation test I.: 0, 125, 250, 500, 1000, 2000 µg/plate
Preincubation test II.: 0, 18.75, 37.5, 75, 150, 300 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: acetone
- Justification for choice of solvent/vehicle: limited solubility of the test substance in water

Controls

Untreated negative controls:

yes

Remarks:

sterility control

Negative solvent / vehicle controls:

yes

Remarks:

Acetone

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: 4-nitroquinoline-N-oxide

Details on test system and experimental conditions:

METHOD OF APPLICATION: standard plate test

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 3


METHOD OF APPLICATION: preincubation

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

NUMBER OF REPLICATIONS: 3

Evaluation criteria:

Doubling of the spontaneous mutation rate, dose-response relationship, reproducibility of the results.

Results and discussion

Test results

Test results 1

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:

Bacteriotoxic effects were observed in the SPT depending on strain and concentrations from 2500 µg/plate onward. In the PIT bacteriotoxicity was observed depending an strain and concentrations from 150 µg-300 µg/plate onward.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Bacteriotoxic effects were observed in the SPT depending on strain and concentrations from 2500 µg/plate onward. In the PIT bacteriotoxicity was observed depending an strain and concentrations from 150 µg-300 µg/plate onward.

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.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Standart plate test (SPT):

Dose (µg/plate)

TA1535

TA100

TA1537

TA98

E. coli WP2 uvrA

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

Acetone

26±2

21±1

132±10

133±19

10±1

17±2

35±2

37±3

37±2

30±2

20

23±3

18±3

130±6

134±9

13±2

10±2

30±3

33±6

30±4

27±1

100

18±3

23±3

147±31

133±29

11±6

9±4

27±5

35±3

25±5

23±5

500

21±3

23±7

119±14

137±21

11±1

8±2

30±4

32±3

20±3

24±2

2500

20±3

27±8x

74±21x

1337

5±1x

3±1x

4±4x

20±1x

22±4

23±3

5000

16±2x

21±6x

47±22x

127±8x

4±2x

-

-

12±2x

15±3

8±3

2 -AA

-

416±120

-

1479±178

-

244±43

-

1065±117

-

240±49

MNNG

1559±240

-

1298±145

-

-

-

-

-

-

-

AAC

-

-

-

-

620±16

-

-

-

-

-

NOPD

-

-

-

-

-

-

944±32

-

-

-

4NQO

-

-

-

-

-

-

-

-

1162±110

-

Mean ± SD

Preincubation-test (PIT) I:

Dose (µg/plate)

TA1535

TA100

TA1537

TA98

E. coli WP2 uvrA

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

Acetone

22±2

20±4

113±6

115±12

10±1

12±1

28±3

45±2

30±4

26±8

125

18±5

14±4

126±8

111±5

10±3

10±2

30±3

36±1

31±3

23±2

250

3±2

18±3

107±9

90±23

10±1

8±1

29±6

32±6

26±2

22±4

500

x

x

x

x

x

x

x

x

x

x

1000

x

x

x

x

x

x

x

x

x

x

2000

x

x

x

x

x

x

x

x

x

x

2-AA

-

132±19

-

647±45

-

115±13

-

549±25

-

234±26

MNNG

729±56

-

818±55

-

-

-

-

-

-

-

AAC

-

-

-

-

334±16

-

-

-

-

-

NOPD

-

-

-

-

-

-

799±46

-

-

-

4NQO

-

-

-

-

-

-

-

-

708±70

-

Preincubation-test (PIT) II:

Dose (µg/plate)

TA1535

TA100

TA1537

TA98

E. coli WP2 uvrA

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

Acetone

21±4

20±3

102±2

115±13

11±3

11±2

28±3

35±3

33±6

34±3

18.75

19±2

17±1

99±4

112±3

11±2

9±2

21±1

35±5

31±1

29±2

37.5

17±3

16±2

99±8

100±8

10±2

10±2

23±3

33±7

30±3

24±2

75

17±1

15±4

84±9

98±15

6±4

7±2

22±2

23±2

23±3

21±4

150

13±1

10±1

90±5

85±16

6±2

6±1

17±6

22±2

23±2

15±3

300

6±1

7±3x

70±2

64±15

2±1

4±2x

11±2

15±3

15±4

9±2x

2-AA

-

110±3

-

569±14

-

94±5

-

518±171

-

203±8

MNNG

652±17

-

857±77

-

-

-

-

-

-

-

AAC

-

-

-

-

404±19

-

-

-

-

-

NOPD

-

-

-

-

-

-

945±50

-

-

-

4NQO

-

-

-

-

-

-

-

-

527±24

-

Mean ± SD

X: reduced background growth

2-AA: 2-aminoanthracene

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

4NQO: 4-nitroquinoline-N-oxide

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 control gave the expected values.