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EC number: 258-887-7 | CAS number: 53956-04-0
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Genetic toxicity: in vitro
Administrative data
- 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:
- test procedure in accordance with national standard methods
Data source
Reference
- Reference Type:
- publication
- Title:
- Unnamed
- Year:
- 2 009
Materials and methods
Test guideline
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- not specified
- Principles of method if other than guideline:
- AMES test was conducted in two independent experiments, plate incorporation test and preincubation test, each carried out without and with metabolic activation (a microsomal preparation derived from
Aroclor 1254-induced rat liver).
The genotypes of the test strains were confirmed as histidine and biotin requirement, (rfa-) deep rough character, UV-sensitivity, and ampicillin-resistance.
For the plate incorporation experiments, the test components were prepared as follows : 0.1 mL of Salmonella cell suspension containing approximately 1x108 viable cells in the late exponential or early stationary phase, 0.1 mL of MGL test solution, or positive control solvent or solvent alone, and 0.5 mL of S9 mix or phosphate buffer were added into 2 mL of soft agar containing 50 mM L-histidine HCl and 50 mM biotin held at 45°C.
The test components were mixed by vortexing the soft agar for 3 seconds at low speed and then poured onto a coded minimal glucose agar plate.
In the preincubation experiments, the test solution of Monoammonium Glycyrrhizinate was preincubated with the test strain solution containing approximately 1x108 viable cells and sterile buffer or the metabolic activation system for 20 minutes at 37°C prior to mixing the overlay agar and poured onto the surface of a coded minimal agar plate.
After 48 to 72 hours incubation of inverted plates in a dark 37°C incubator, the revertant colonies on the test plates and on the control plates were counted, and the presence of the background lawn on all plates was confirmed. Triplicate plates were examined at each concentration level and strain. - GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- Ammonium glycyrrhizate
- EC Number:
- 258-887-7
- EC Name:
- Ammonium glycyrrhizate
- Cas Number:
- 53956-04-0
- Molecular formula:
- C42H65NO16
- IUPAC Name:
- ammonium glycyrrhizate
Constituent 1
- Specific details on test material used for the study:
- The appearance and properties were a white yellowish-white odorless powder with mp=209°C. MGL was dissolved in 0.9% NaCl solution or dimethyl sulfoxide(DMSO).
Method
Species / strain
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 fraction (post-mitochondrial fraction) was prepared from rats treated with Aroclor 1254, collected and stored in liquid nitrogen. S9 mix (5% S9) was freshly prepared on the day of the test.
- Test concentrations with justification for top dose:
- As no signs of cytotoxicity were noted at up to the top concentration of 5000µg/plate in the preliminary test without metabolic activation in test strain TA 100 employing a plate incorporation test, five concentrations separated by a half-log interval ranging from 100 to 5000µg/plate were employed in the independent experiments without and with metabolic activation.
- Vehicle / solvent:
- MGL was dissolved in 0.9% NaCl solution or dimethyl sulfoxide (DMSO).
Controls
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- 2-nitrofluorene
- sodium azide
- cyclophosphamide
- methylmethanesulfonate
- other: 2-amino-anthracene (with metabolic activation, for TA 98, TA 102, and TA 1537 strains)
- Details on test system and experimental conditions:
- AMES test was conducted in two independent experiments, plate incorporation test and preincubation test, each carried out without and with metabolic activation (a microsomal preparation derived from
Aroclor 1254-induced rat liver).
The genotypes of the test strains were confirmed as histidine and biotin requirement, (rfa-) deep rough character, UV-sensitivity, and ampicillin-resistance.
For the plate incorporation experiments, the test components were prepared as follows : 0.1 mL of Salmonella cell suspension containing approximately 1x108 viable cells in the late exponential or early stationary phase, 0.1 mL of MGL test solution, or positive control solvent or solvent alone, and 0.5 mL of S9 mix or phosphate buffer were added into 2 mL of soft agar containing 50 mM L-histidine HCl and 50 mM biotin held at 45°C.
The test components were mixed by vortexing the soft agar for 3 seconds at low speed and then poured onto a coded minimal glucose agar plate.
In the preincubation experiments, the test solution of MGL was preincubated with the test strain solution containing approximately 1x108 viable cells and sterile buffer or the metabolic activation system for 20 minutes at 37°C prior to mixing the overlay agar and poured onto the surface of a coded minimal agar plate.
After 48 to 72 hours incubation of inverted plates in a dark 37°C incubator, the revertant colonies on the test plates and on the control plates were counted, and the presence of the background lawn on all plates was confirmed. Triplicate plates were examined at each concentration level and strain. - Evaluation criteria:
- Since the statistical evaluation of the results of the AMES test is still under discussion, and the range of spontaneous reversion frequencies of TA 98, 100, 102, 1535, and 1537 in our test circumstance are generally 20- 60, 100-200, 240-320, 10-35, and 3-20, respectively, the test item would be considered to show a positive response if the results were satisfied with the all following criteria : i) if the number of revertants would be significantly increased at p< 0.05 by U-test according to MANN and WHITNEY compared with the solvent control,
to at least 2-fold of the solvent control for TA 98, TA 100 and TA 102, and 3-
fold of the solvent control for TA 1535 and TA 1537 in both independent experiments, ii) if a significant concentration(log value) - related effect at p< 0.05 by Spearman’s rank correlation coefficient would be observed, and iii) if positive results would have to be reproducible and the histidine independence of the revertants would have to be confirmed by streaking random samples on histidine-free agar plates. Cytotoxicity was defined as a reduction in the number of colonies by more than 50% compared with the solvent control and/or a scarce background lawn. - Statistics:
- Since the statistical evaluation of the results of the AMES test is still under discussion, and the range of spontaneous reversion frequencies of TA 98, 100, 102, 1535, and 1537 in our test circumstance are generally 20- 60, 100-200, 240-320, 10-35, and 3-20, respectively, the test item would be considered to show a positive response if the results were satisfied with the all following criteria : i) if the number of revertants would be significantly increased at p< 0.05 by U-test according to MANN and WHITNEY compared with the solvent control,
to at least 2-fold of the solvent control for TA 98, TA 100 and TA 102, and 3-
fold of the solvent control for TA 1535 and TA 1537 in both independent experiments, ii) if a significant concentration(log value) - related effect at p< 0.05 by Spearman’s rank correlation coefficient would be observed, and iii) if positive results would have to be reproducible and the histidine independence of the revertants would have to be confirmed by streaking random samples on histidine-free agar plates. Cytotoxicity was defined as a reduction in the number of colonies by more than 50% compared with the solvent control and/or a scarce background lawn.
Results and discussion
Test results
- Key result
- Species / strain:
- other: All strains
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- other: Solvent control is used as negative control
- Positive controls validity:
- valid
Applicant's summary and conclusion
- Conclusions:
- The effect of MGL on the bacterial reverse mutation activity using 5 Salmonella typhimurium stains TA 98, TA 100, TA 102, TA 1535 and TA 1537 has been addressed in two independent experiments, each carried out without and with metabolic activation by a rat post-mitochondorial fraction(S9 mix) from Aroclor 1254 induced animals. The first experiment was carried out as a plate incorporation and the second as a preincubation test.
In the result of the preliminary cytotoxicity test, no signs of cytotoxicity were noted in the plate incorporation test of TA 100 without metabolic activation at up to the top concentration of 5000 µg/plate. Hence, 5000 µg/ plate was chosen as the top concentration for the main study. While there were no signs of cytotoxicity noted in the plate incorporation test without and with metabolic activation and in the preincubation test with metabolic activation, cyototoxicity was observed in the preincubation test without metabolic activation at the top concentration of 5000 µg/plate in all test strains. On the other hand, no mutagenic effect was observed for MGL tested at up to 5000 µg/plate in any of the 5 test strains in two independent experiments without and with metabolic activation, respectively. Consequently, under the present test conditions MGL tested at up to a cytotoxic concentration caused no mutagenic effect in the Salmonella typhimurium strains TA 98, TA 100, TA 102, TA 1535, and TA 1537 neither in the plate incorporation test nor in the preincubation test each carried out without and with metabolic
activation.
Under tests conditions of GLP study, results are considered scientifically valid to conclude on absence of mutagenic activity on prokaryotic cells.
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