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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
Study period:
1982-01-26 to 1982-02-23
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
However, no E. coli strain or S. typhimurium TA102 was included. Limited information was provided on the test substance.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1982
Report Date:
1982

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
No E. coli strain or S. typhimurium TA102 were included in the study. Limited information was provided on the test substance.
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
- Physical state: liquid
- Storage condition of test material: Room temperature
Specific details on test material used for the study:
- Name of test material (as cited in study report): Poly[oxy(methyl-1,2-ethanediyl)], alpha-(2-aminomethylethyl)-omega-(2-aminomethylethoxy)
- Substance type: active
- Physical state: clear, colorless liquid
- Stability under test conditions: There was no apparent change in the physical state of the test substance during the assay.
- Test article 4236-44-15; Order #J-127.

Method

Target gene:
histidine locus
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell type (if applicable):
N/A
Additional strain / cell type characteristics:
other: See table below.
Species / strain / cell type:
S. typhimurium TA 1538
Details on mammalian cell type (if applicable):
N/A
Additional strain / cell type characteristics:
other: See table below.
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254 induced rat liver S-9 was used as the metabolic activation. There were 0.6 ml of S-9 supernatant (47 mg protein per ml) per 1.0 ml of S-9 mix used in the rat liver microsomal activation system.
Test concentrations with justification for top dose:
100, 333, 1000, 3333 and 10,000 ug/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: distilled water
- Justification for choice of solvent/vehicle: The test substance was miscible in the solvent.
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
distilled water (test substance vehicle)
True negative controls:
no
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
without metabolic activation Migrated to IUCLID6: 1 ug/plate for TA1535 and TA100
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
distilled water (test substance vehicle)
True negative controls:
no
Positive controls:
yes
Positive control substance:
2-nitrofluorene
Remarks:
without metabolic activation Migrated to IUCLID6: 5 ug/plate for TA1538 and TA98
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
distilled water (test substance vehicle)
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
without metabolic activation Migrated to IUCLID6: 150 ug/plate for TA1537
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
distilled water (test substance vehicle)
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-anthramine-5 ug/plate for all strains
Remarks:
with metabolic activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation): Top agar, used as an overlay, was reconstituted into a molten state and supplemented with 0.5 mM histidine-0.5 mM Biotin at a volume of 0.1 ml per ml of agar, and maintained at 45 degrees C until used. Using sterile technique, the following were added to tubes in the following order: 2 ml aliquots of top agar solution, 0.1 ml of tester strain, and 0.1 ml of the appropriate concentration of the test substance. S-9 was added in addition when appropriate. The tubes were vortexed and poured onto minimal glucose plates. The samples was evenly distributed on the plate, and the top agar overlay was allowed to harden.

DURATION
- Exposure duration: 48-72 hours
- Selection time (if incubation with a selection agent): 48-72 hours (simultaneous with exposure)


SELECTION AGENT (mutation assays): histidine


NUMBER OF REPLICATIONS: All solvent and positive controls were plated in triplicate, while all test substance groups were plated in duplicate.


DETERMINATION OF CYTOTOXICITY
- Method: bacterial background lawn

Evaluation criteria:
A positive result was defined as a reproducible, dose related increase in the number of histidine-independent colonies. If the solvent control was within one standard deviation of the historical mean for control values and the test substance produced the highest increase equal to or greater than three times the solvent control value, the test substance was considered mutagenic. A negative result was defined as the absence of a reproducible increase in the number of histidine-independent colonies.
Statistics:
N/A

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:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
RANGE-FINDING/SCREENING STUDIES: a preliminary toxicity test was performed with S. typhimurium TA1538 and TA100 using dose levels of 100-10,000 ug/plate in the absence of metabolic activation using the plate incorporation assay.. All solvent and test substance groups were performed in duplicate. Plates were incubated for 48 hours following which the background lawn and spontaneous revertants were observed and scored as normal growth, inhibited growth or no growth. Inhibition was scored by the presence of pindot colonies and the absence of a confluent lawn of bacteria. The concentration giving moderate inhibition of bacterial lawn growth served as the high dose in the plate incorporation assay. If not toxicity was produced, the high dose for the test substance was prepared at 100 mg/ml. Lower doses would be at log dilutions.
No inhibition at the dose levels tested was observed and therefore, 10,000 ug/plate was chosen as the high dose for the main assay.

COMPARISON WITH HISTORICAL CONTROL DATA: All solvent and positive controls were within the acceptable range of mean historical data. The results for the test substance were negative.

Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Substance

ug/plate

TA1535

TA1537

TA1538

TA98

TA100

 

-

+

-

+

-

+

-

+

-

+

Solvent

 

12

14

19

21

23

29

31

40

123

103

Sodium Azide

1

601

 

 

 

 

 

 

 

736

 

2-Nitrofluorene

5

 

 

 

 

1136

 

931

 

 

 

9-aminoacridine

150

 

 

2295

 

 

 

 

 

 

 

2-anthramine

5

 

182

 

465

 

2088

 

2832

 

1001

Test Substance

10000

13

10

22

23

18

35

42

29

78

106

3333

15

11

19

23

21

18

46

43

113

110

1000

19

14

21

28

20

22

44

43

117

102

333

15

18

17

24

16

27

40

35

119

117

100

17

16

18

24

24

27

39

29

111

119

 

Applicant's summary and conclusion

Conclusions:
The test substance was evaluated for mutagenic activity in the Ames Salmonella typhimurium Assay with and without metabolic activation. Based on the results, the test substance was determined to be non-mutagenic.