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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
Study period:
5 November 1982 to 18 November 1982
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guidelines and the study was conducted under GLP conditions.

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
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
9-Octadecenoic acid (Z)-, monoester with 1,2,3-propanetriol ester with boric acid (H3BO3)
EC Number:
264-092-6
EC Name:
9-Octadecenoic acid (Z)-, monoester with 1,2,3-propanetriol ester with boric acid (H3BO3)
Cas Number:
63310-16-7
Molecular formula:
C21H39O5B (based on the representative structure below)
IUPAC Name:
{2-hydroxy-3-[(9Z)-octadec-9-enoyloxy]propoxy}boronic acid 2-hydroxy-3-{[hydroxy({2-hydroxy-3-[(9Z)-octadec-9-enoyloxy]propoxy})boranyl]oxy}propyl (9Z)-octadec-9-enoate 3-{[bis({2-hydroxy-3-[(9Z)-octadec-9-enoyloxy]propoxy})boranyl]oxy}-2-hydroxypropyl (9Z)-octadec-9-enoate
Test material form:
not specified
Details on test material:
- Physical state: liquid

Method

Target gene:
Histidine synthesis
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
not applicable
Species / strain / cell type:
S. typhimurium TA 1538
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
S9 mix
Test concentrations with justification for top dose:
0.1, 0.3, 1.0, 3.0 and 10.0 mg/plate both in the presence and absence of metabolic activation
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
sodium azide
other: 2-aminoanthracene
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation). Top agar containing the test material, bacteria, and in some cases S9 mix was mixed and uniformly distributed over the surface of a minimal agar plate. Prior to testing, the molten top agar was prepared by adding 9 x 10^-5 mmoles each of L-histidine and D-biotin. 0.5 mL S9 mix (or S9 buffer) was then added to the number of aliquots of one strain required for one concentration, followed by 0.1 mL of the appropriate concentration of the test substance preparation. Finally 2 mL top agar was then added to each aliquot (2.5 mL for tests without S9 mix), and the resulting mixture poured onto the surface of a prepared pre-labelled plate.

DURATION
- Exposure duration: Plates were incubated inverted at 37 °C for two to three days

NUMBER OF REPLICATIONS: Test concentrations were performed in triplicate

- EVALUATION PROCEDURE: Following the total incubation the number of revertant colonies was counted and the presence and condition of the bacterial lawn was noted.
Evaluation criteria:
Criteria for response in the Ames test are outlined in the field "Any other information on materials and methods incl. tables" section.

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:
not applicable
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
RANGE-FINDING/SCREENING STUDIES:
The test material was checked for toxicity on strain TA100 without S9 mix at dose levels of 0.005 to 10 mg/plate. The plates were run in duplicate. No toxicity was observed at these dose levels and so it was decided to test concentrations of test material from 0.1 to 10.0 mg/plate in the definitive study.

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

Any other information on results incl. tables

Table 1: Results (Mean values of triplicate plates)

Treatment

Amount/plate

No. revertant colonies per plate

TA98

TA100

TA1535

TA1537

TA1538

without S9

with S9

without S9

with S9

without S9

with S9

without S9

with S9

without S9

with S9

Test material

10.0 mg

16

35

85

118

25

15

15

18

14

20

3.0 mg

25

32

86

102

24

11

16

19

14

23

1.0 mg

14

36

94

100

26

14

14

18

16

24

0.3 mg

22

32

113

103

25

13

15

22

21

23

0.1 mg

22

32

88

85

30

10

18

19

14

23

DMSO

0.1 mL

19

33

90

90

23

14

21

20

13

23

Positive control

2-nitrofluorene (10 µg)

2-aminoanthracene (2 µg)

sodium azide (1 µg)

2-aminoanthracene (2 µg)

sodium azide (1 µg)

2-aminoanthracene (2 µg)

9-aminoacridine (50 µg)

2-aminoanthracene (2 µg)

2-nitrofluorene (10 µg)

2-aminoanthracene (2 µg)

1964

3765

781

3605

589

254

1300

385

3928

4256

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

Under the specific conditions of this assay, the test material gave a negative (i.e. non-mutagenic), response in Salmonella typhimurium strains TA1535, TA1537, TA1538, TA98 and TA100 both in the presence and absence of metabolic activation. No toxicity was observed. The tester strains responded to the positive controls, known mutagens, as expected, confirming the validity of the test method employed. The study is considered to be reliable, relevant and adequate for risk assessment and classification and labelling purposes.
Executive summary:

The potential of the test material to cause gene mutation in bacterial strains was determined following a methodology equivalent to standardised guideline OECD 471. Five strains of Salmonella typhimurium (TA1535, TA1537, TA1538, TA98 and TA100) were treated in the presence and absence of a rat liver derived metabolic activation system (S9 mix). Under the conditions of the study, the test material did not induce any significant, reproducible increases in the observed number of revertant colonies in any of the tester strains used either in the presence or absence of metabolic activation. No toxicity was observed. The tester strains responded to the positive controls, known mutagens, as expected, confirming the validity of the test method employed.