Extract from the seeds of Trigonella foenum-graecum (Fabaceae) obtained by extraction with polar solvents

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

According to the test carried out under the TG OECD 471 and in GLP conditions, the test substance does not have any mutagenic potential.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

Identification/Synonym FENUGREEK EXTRACT
Substance type UVCB substance
EC No. 950-727-0
Recommended storage Ambient (18-20°C) and protected from temperatures below 4°C; dry (< 70% rh);
conditions to be stored under dark conditions

Target gene:

Tryptophan and histidine

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:

Type and composition of metabolic activation system:
- Rat (Sprague Dawley)
- Mixing of S9 tissue fraction and cofactors
- quality controls of S9
- enzymatic activity
- sterility
- metabolic capability

Test concentrations with justification for top dose:

TOXICITY TEST WITH AND WITHOUT METABOLIC ACTIVATION:
5000; 1580; 500; 158 and 50.0 μg/plate

Main Assay I and II
5000; 2500; 1250; 625 and 313 μg/plate

Vehicle / solvent:

Sterile water and Dimethylsulfoxide (DMSO)

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

methylmethanesulfonate

other: 2-aminoanthracene

Details on test system and experimental conditions:

NUMBER OF REPLICATIONS:
- Preliminary toxicity test one plate for each concentration with and without S9
- Main assay I and II
- negative and positive controls with and without S9
- Three replicate plates per test point

METHOD OF TREATMENT/ EXPOSURE:
- Cell density at seeding: bacteria (1-5 x 10^8 cells / mL).

TREATMENT AND HARVEST SCHEDULE:
- Main Assay II: Preincubation period, 37°C for 30 min
- Exposure duration: 72 hours at 37°C

METHODS FOR MEASUREMENTS OF GENOTOXICIY

Plate count (number of colonies)

Rationale for test conditions:

Guideline study

Evaluation criteria:

For the test item to be considered mutagenic, two-fold (or more) increases in mean revertant numbers must be observed at two consecutive dose levels or at the highest practicable dose level only. In addition there must be evidence of a dose-response relationship showing increasing numbers of mutant colonies with increasing dose levels.

Statistics:

Regression lines
Correlation co-efficient (r)
Students "t" Test

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not applicable

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not applicable

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not applicable

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not applicable

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility: 50.0 mg/mL (maximum concentration of 5000 µg/plate)

RANGE-FINDING/SCREENING STUDIES:
Toxicity test: 5000; 1580; 500; 158 and 50.0 μg/plate
Main assay I and II: 5000; 2500; 1250; 625 and 313 μg/plate

Conclusions:

It is concluded that the test item does not induce reverse mutation in Salmonella typhimurium or Escherichia coli in the absence or presence of S9 metabolism, under the reported experimental conditions.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)