α-d-Glucopyranosiduronic acid, (3β,20β)-20-carboxy-11-oxo-30-norolean-12-en-3-yl 2-O-β-d-glucopyranuronosyl-, dipotassium salt

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16/01/2002-18/01/2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

α-d-Glucopyranosiduronic acid, (3β,20β)-20-carboxy-11-oxo-30-norolean-12-en-3-yl 2-O-β-d-glucopyranuronosyl-, dipotassium salt

EC Number:

272-296-1

EC Name:

α-d-Glucopyranosiduronic acid, (3β,20β)-20-carboxy-11-oxo-30-norolean-12-en-3-yl 2-O-β-d-glucopyranuronosyl-, dipotassium salt

Cas Number:

68797-35-3

Molecular formula:

C42H62O16.2K

IUPAC Name:

α-d-Glucopyranosiduronic acid, (3β,20β)-20-carboxy-11-oxo-30-norolean-12-en-3-yl 2-O-β-d-glucopyranuronosyl-, dipotassium salt

Specific details on test material used for the study:

Preparation : By mixing 300 mg of the test substance with 6 mL of distilled water, the highest concentration (5000 µg / 0.1 µL / plate) was prepared. This solution was serially diluted to half its concentration to prepare the others concentrations.

Method

Target gene:

Not specified

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:

S9mix (from Sprague dawley rats + 9 mL of distilled water contaiing S9 cofactors)

Test concentrations with justification for top dose:

312.5 ; 625 ; 1250 ; 2500 ; 5000 µg/0.1 µL/plate

Vehicle / solvent:

distilled water

Controls

Untreated negative controls:

yes

Remarks:

Distilled water

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

9-aminoacridine

sodium azide

other: 2-(2-furyk)-3-(5-nitro-2furyl) acrylamide (AF2) and 2-aminoanthracène (2-AA)

Details on test system and experimental conditions:

Culture media :
- Nutrient broth
Nutrient broth No.2 was dissolved in distilled water to prepare a 2.5% culture medium and 20mL of the medium added in each Erlenmeyer flask was sterilized for 15 min. at 121°C.
- Minimum glucose agar medium
TESMEDIA AN (Wako Pure Chemical Industries) was used. Each plate was marked by writing an ID number on the top of the rid.
- Top agar
For four Salmonella typhimurium strains, 10-part of 0.6 % agar containing 0.5 % Na i, 1-part of 0.5 mM D-biotin and. 1-part of 0.5 mM L-histidine were mixed.
For Escherichia coli; 10-part of agar and 1-part of 0.5mM L-tryptophan were mixed

Evaluation criteria:

Assessment
Positive reaction (+) : The number of revertant colonied per plate was more than twice that for the negative control group and this increased number of revertant colonies was reproducible and dose-dependent.
Negative reaction (—) : Other reactions
Growth inhibition : The background lawn for the test substance group was clearly rougher or clearer compared to that of the negative control group.

Statistics:

Not specified

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not applicable

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not applicable

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not applicable

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not applicable

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Not specified

Applicant's summary and conclusion

Executive summary:

Based on these findings, it was concluded that under the conditions established for the present study, DIPOTASSIUM GLYCYRRHIZINATE was judged as non-mutagen.