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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: test performed according to OECD guideline and GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008
Report Date:
2008

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
GLP compliance:
yes (incl. certificate)
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): Vegeflux soy
- Substance type: UVCB
- Physical state: viscous liquid
- Stability under test conditions: data not available
- Storage condition of test material: at room temperature

Method

Species / strain
Species / strain / cell type:
S. typhimurium, other: TA 98, TA 100, TA 102, TA 1535, TA 1537
Metabolic activation:
with and without
Metabolic activation system:
from rat liver (no other information available)
Test concentrations with justification for top dose:
5, 1.58, 0.5, 0.158, and 0.05 µL per plate
Vehicle / solvent:
DMSO
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: see detail in table 1
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 hours at 37°C

NUMBER OF REPLICATES: three plates per concentration

OTHER: the revertant colonies were scored from each plate by visual counting of the colonies

Evaluation criteria:
Mutagenic response was considered to be positive, if the mean number of His+ revertant colonies double or more than double the number obtained with solvent (negative) control at any dose, and in any tester strain, with or without metabolic activation.
Statistics:
For statistical analysis the data of mean revertant colony frequencies were compared by Student 't' test.

Results and discussion

Test results
Species / strain:
S. typhimurium, other: TA 98, TA 100, TA 102, TA 1535, TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH, Effects of osmolality, Evaporation from medium, Water solubility: data not available
- Precipitation: the test item did not precipitate the top agar even at the highest dose tested

RANGE-FINDING/SCREENING STUDIES:
Prior to the commencement of the main study the test item was assessed for toxicity using TA 100 strain. Five concentrations of the test item such as 5, 1.58, 0.5, 0.158 and 0.05 µL/plate were prepared and tested for bacterial toxicity (preliminary test).
No inhibition of background law of auxotrophic, His- cells and spontaneous number of His+ cells of the tester strain was noticed even at the highest dose of 5 µL/plate ; hence 5 µL was selected as the highest dose for mutagenicity testing.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table 2: Results with Vegeflux soy – Summary

Test item

Test concentration per plate

S9 mix

Average Histidine revertant colonies/plate

Base par types

Frame shift types

TA 100

TA 102

TA 1535

TA 1537

TA 98

Negative control

100 µL

-

109.67

223.67

23.33

9.67

23.00

+

113.67

227.33

23.33

11.67

23.67

Vegeflux soy

5

 

 

-

110.33

222.33

26.67

13.67

23.67

1.58

108.33

231.00

25.00

14.00

23.00

0.5

108.00

226.00

23.33

10.00

23.67

0.158

116.00

231.00

26.33

14.67

25.33

0.05

116.00

217.00

22.33

9.33

24.67

Vegeflux soy

5

 

 

+

117.00

228.67

22.67

13.33

22.67

1.58

112.33

228.67

25.33

11.67

29.00

0.5

115.67

232.67

24.67

10.00

26.00

0.158

114.00

221.00

24.67

10.33

25.67

0.05

112.33

226.67

23.33

13.00

25.33

Mitomycin c

0.5

-

NA

602.33

NA

NA

NA

Sodium azide

5

-

713.00

NA

691.33

NA

NA

2-nitrofluorene

10

-

NA

NA

NA

NA

870.67

2-aminoacridine

50

-

NA

NA

NA

272.33

NA

2-amino anthracene

1

+

666.00

610.67

687.33

257.67

871.67

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative with and witout metabolic activation

Vegeflux soy is non-mutagenic by Ames bacterial reverse mutation assay under the condition of the test employed.
Executive summary:

In a reverse gene mutation assay in bacteria (Venkatasubramani R, 2008), strains TA 98, TA 100, TA 102, TA 1535, TA 1537 of S. typhimurium were exposed toVegeflux soy diluted in DMSO at concentrations of 5, 1.58, 0.5, 0.158, and 0.05 µL per plate in the presence and absence of mammalian metabolic activation. The direct plate incorporation method was used. The test item was tested up to limit concentration.

 

Vegeflux soy did not induce mutagenic response in any of the five tester strains up to the highest concentration. There was no evidence of induced mutant colonies over background. The positive controls induced the appropriate responses in the corresponding strains.

 

This study is classified as acceptable. This study satisfies the requirement for Test Guideline OECD 471 for in vitro mutagenicity (bacterial reverse gene mutation) data.