Registration Dossier
Registration Dossier
Data platform availability banner - registered substances factsheets
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Diss Factsheets
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 283-415-1 | CAS number: 84625-40-1 Extractives and their physically modified derivatives such as tinctures, concretes, absolutes, essential oils, oleoresins, terpenes, terpene-free fractions, distillates, residues, etc., obtained from Trigonella foenum-graecum, Leguminosae.
- 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
- Study period:
- 03 January to 15 January 2018
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 018
- Report date:
- 2018
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- Principles of method if other than guideline:
- Not applicable
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- Fenugreek, ext.
- EC Number:
- 283-415-1
- EC Name:
- Fenugreek, ext.
- Cas Number:
- 84625-40-1
- Molecular formula:
- Not applicable (UVCB)
- IUPAC Name:
- Absolute of Trigonella foenum graecum L. (Leguminosae) obtained from seeds by organic solvent treatment and subsequent ethanol extraction
- Test material form:
- liquid: viscous
- Remarks:
- Brown-yellow viscous liquid
- Details on test material:
- Test item: Fenugreek Absolute
Batch No.: 2709379
Identity: 100 % UVCB Substance
Appearance: Brown-yellow viscous liquid
Storage conditions: Ambient temperature (10 °C to 30 °C), in original aluminium bottle, dark
Expiry date: October 24, 2018
Constituent 1
Method
- Target gene:
- Histidine and tryptophan for S. typhimurium and E. coli, respectively.
Species / strain
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
- Details on mammalian cell type (if applicable):
- No applicable
- Additional strain / cell type characteristics:
- not applicable
- Metabolic activation:
- with and without
- Metabolic activation system:
- Obtained from Molecular Toxicology Incorporated, Boone, NC, USA (MolTox™) and prepared according to the method of Ames et al from male Sprague Dawley rats induced with a single dose of Aroclor 1254.
- Test concentrations with justification for top dose:
- In order to determine the toxicity of Fenugreek Absolute to the test bacteria, an initial plate incorporation experiment was carried out using TA98 and WP2 uvrA in the presence and absence of S 9 mix. The dose levels used were 1.5, 5, 15, 50, 150, 500, 1500 and 5000 µg/plate. Based on the range finder observations, dose levels for the main experiment were chosen to include the guideline regulatory maximum dose level of 5000 µg/plate.
- Vehicle / solvent:
- Dimethylsulphoxide (DMSO), a commonly used vehicle for which extensive historical control data is available; at concentrations up to 50 mg/mL.
Controlsopen allclose all
- Untreated negative controls:
- yes
- Remarks:
- untreated
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 4-nitroquinoline-N-oxide
- 9-aminoacridine
- 2-nitrofluorene
- sodium azide
- Remarks:
- Without S9
- Untreated negative controls:
- yes
- Remarks:
- untreated
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-Aminoanthracene (2AA)
- Details on test system and experimental conditions:
- Salmonella typhimurium and Escherichia coli strains were obtained from Molecular Toxicology Incorporated, Boone, NC, USA (MolTox™) and used in the Bacterial Reverse Mutation test.
The Salmonella typhimurium and Escherichia coli strains are defective in DNA repair (Δuvr B , A- respectively); this confers extra sensitivity to DNA damage. The Salmonella strains have a defective lipopolysaccharide barrier on the cell wall (rfa-) which affords greater permeability to large molecules. The strains TA98 and TA100 also contain a plasmid (pKM101) which enhances error prone repair and confers ampicillin resistance.
The strains are tested routinely for:
1. Auxotrophic control of growth, (histidine or tryptophan dependence).
2. Deep rough mutation (rfa) of the bacterial cell wall (cell membrane permeability, crystal violet sensitivity).
3. Plasmid presence (Ampicillin resistance).
4. Absence of uvrA or uvrB repair enzyme systems (sensitivity to UV irradiation).
TA1535, TA100 and WP2 uvrA are reverted to prototrophy by base substitution mutagens. TA1537 and TA98 are reverted by frame shift mutagens; TA100 can also be affected by these on occasion.
Before the experiment, 25 mL quantities of nutrient broth were inoculated with 100 µL from a freshly thawed vial of the appropriate strain, and incubated overnight at 37 °C to give a pure culture at approximately 10^9 cells per mL. - Rationale for test conditions:
- Range finder: As all the Salmonella strains and the Escherichia strain show a similar toxic response to most chemicals, only one strain of each species, typically TA98 and WP2 uvrA, was tested by plate incorporation in this experiment.
A range of concentrations up to a maximum of 5000 μg/plate was used, with duplicate plates at each concentration, both with and without S-9.
The plates examined for integrity of the background lawns after approximately 48 hours ± 1 hour incubation.
Experiment I: Mutation experiment, plate incorporation : The mutation experiment was carried out using fresh bacterial cultures for each experiment.
The test item was tested at sufficient concentration levels to provide at least 5 analysable concentrations, with triplicate Petri dishes prepared for each control and experimental point. The following was added to sterile disposable tubes containing 2 mL of L-histidine: D-biotin (Salmonella strains) or L-tryptophan (Escherichia strain) supplemented top agar. The contents of each tube was mixed and added to a Petri dish containing minimal agar. When the top agar has set, the Petri dishes will be inverted and incubated at 37 °C ± 3 °C for approximately 66 hours ± 1 hour.
Experiment II: Mutation experiment, preincubation: The mutation experiment was carried out using fresh bacterial cultures for each experiment.
The test item was tested at sufficient concentration levels to provide 5 analysable concentrations, with triplicate Petri dishes prepared for each control and experimental point. Each tube was fitted with a sterile cap and incubated at approximately 37 °C with shaking, for 20 minutes.
At the end of this period, 2 mL of top agar (L-histidine: D-biotin for Salmonella strains or L-tryptophan for Escherichia strain) was added to each tube, the contents mixed and added to Petri dishes containing minimal agar. When the top agar has set, the Petri dishes are inverted and incubated at 37 °C ± 3 °C for approximately 66 hours ± 1 hour. - Evaluation criteria:
- A test was considered to be positive if the test item induced dose related statistically significant increases in numbers of revertants over the range tested and/or a reproducible increase at one or more concentrations in the number of revertant colonies per plate in at least one strain with or without metabolic activation system, compared with negative Controls scored in 2 separate experiments. Biological relevance of the result should be considered. Statistical significance should not be the only determining factor for a positive response.
A test was considered to be negative if the test item produced no greater increases in revertants, than may be expected from normal variation in the negative Control number of revertants, for any strain, in either experiment. - Statistics:
- Mean, standard deviation (s.d.) and Dunnett's t-statistic (t) were calculated for each group and bacterial strain
Results and discussion
Test resultsopen allclose all
- Key result
- Species / strain:
- S. typhimurium TA 1535
- 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:
- valid
- 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 nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- 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 nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- 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 nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- E. coli WP2 uvr A
- 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:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- Toxicity Range finder : No toxicity presents. The only effect is a precipitation at the top dose (5000 µg/plate).
Experiment 1: All negative Controls gave mean counts of spontaneous revertant colonies within expected historical negative Control ranges obtained in this laboratory.
All positive Controls induced marked increases in the number of revertant colonies and gave mean counts of induced revertant colonies which were within expected historical positive Control ranges. The response to the positive Controls demonstrates that the bacteria were sensitive to the mutagens and that the S 9 mix was able to metabolise the pro-mutagen, 2AA, to a mutagen.
Fenugreek Absolute was tested up to the regulatory maximum dose level of 5000 µg/plate in all strains in the presence and absence of S 9 mix, under plate incorporation conditions.
Precipitation was observed at 5000 µg/plate in all strains in the presence and absence of S 9 mix.
Calculated t values were compared with reference critical values and, in the event that the calculated values exceeded the critical values, they were to be highlighted in the appropriate tables, to indicate statistically significant increases in colony counts. None of the calculated t values in this experiment exceeded the critical values.
There were no dose-related or statistically significant increases in revertant numbers observed in any strain at any dose of Fenugreek Absolute, in the presence or absence of S 9 mix, under plate incorporation conditions.
Experiment 2: All negative Controls gave mean counts of spontaneous revertants within expected ranges obtained in this laboratory.
All positive Controls gave mean counts of induced revertants within expected ranges obtained in this laboratory. The response to the positive Controls indicated that the bacteria were sensitive to the mutagens and that the S 9 mix was able to metabolise the pro-mutagen, 2AA, to a mutagen.
Fenugreek Absolute was tested up to the regulatory maximum dose level of 5000 µg/plate in all strains in the presence and absence of S 9 mix, under pre-incubation conditions.
Precipitation was observed at 5000 µg/plate in all strains in the presence and absence of S 9 mix.
Calculated t values were compared with reference critical values and, where the calculated values exceeded the critical values, were highlighted in the appropriate tables, indicating statistically significant increases in colony counts. None of the calculated t values in this experiment exceeded the critical values.
There were no dose-related statistically significant increases in revertant numbers observed in any strain at any dose of Fenugreek Absolute, in the presence or absence of S 9 mix, under pre incubation conditions.
Historical control data: The negative and positive Controls from the mutation experiment were compared with previously established ranges of these parameters for this laboratory. Control data were accepted for this study as they were within the minimum and maximum values recorded within the previous 2 year period.
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: Not reported
- Effects of osmolality: Not reported
- Evaporation from medium: Not reported
- Water solubility: Not reported
- Precipitation: The test item precipitated in the overlay agar at the highest investigated concentration.
- Other confounding effects: Not reported
MUTAGENICITY RESULTS
See 'Any other information on results incl. tables'
Any other information on results incl. tables
Table 1 : Mean Number of Revertants Per Plate – Experiment 1 – Plate Incorporation
Strain |
% S-9 |
Dose level of test item (µg/plate) |
PC |
|||||
0 |
50 |
150 |
500 |
1500 |
5000 |
|||
TA1535 |
0 |
19.7 |
12.7 |
15.3 |
13.7 |
17.0 |
13.0 |
659.7 |
TA1537 |
0 |
9.3 |
5.3 |
7.7 |
6.7 |
2.3 |
5.3 |
681.0 |
TA98 |
0 |
20.0 |
19.0 |
25.0 |
21.3 |
19.7 |
23.7 |
94.7 |
TA100 |
0 |
82.7 |
89.3 |
95.3 |
97.0 |
96.0 |
91.3 |
588.7 |
WP2uvrA |
0 |
18.0 |
19.0 |
21.7 |
22.3 |
22.7 |
22.7 |
552.3 |
Strain |
% S-9 |
Dose level of test item (µg/plate) |
PC |
|||||
0 |
50 |
150 |
500 |
1500 |
5000 |
|||
TA1535 |
10 |
14.7 |
12.0 |
12.7 |
15.0 |
14.3 |
14.3 |
165.0 |
TA1537 |
10 |
10.7 |
9.7 |
10.0 |
6.0 |
8.0 |
8.3 |
183.7 |
TA98 |
10 |
25.3 |
24.7 |
32.7 |
27.7 |
32.0 |
32.7 |
654.7 |
TA100 |
10 |
100.3 |
101.7 |
95.0 |
97.3 |
59.7 |
83.7 |
1697.7 |
WP2uvrA |
10 |
31.3 |
37.3 |
25.3 |
28.7 |
24.3 |
30.7 |
150.3 |
PC: |
positive control |
|||||||
ppt: |
compound precipitation |
Table 2 :Mean Number of Revertants Per Plate – Experiment 2 – Pre-incubation
Strain |
% S-9 |
Dose level of test item (µg/plate) |
PC |
|||||
0 |
50 |
150 |
500 |
1500 |
5000 |
|||
TA1535 |
0 |
13.3 |
14.3 |
12.0 |
14.0 |
14.0 |
13.0 |
527.0 |
TA1537 |
0 |
7.3 |
7.3 |
5.7 |
3.3 |
5.5 |
6.0 |
527.7 |
TA98 |
0 |
16.3 |
18.0 |
17.0 |
19.0 |
17.3 |
21.0 ppt |
123.0 |
TA100 |
0 |
70.7 |
90.3 |
82.0 |
90.7 |
86.0 |
82.0 |
522.3 |
WP2uvrA |
0 |
28.3 |
25.3 |
25.0 |
30.7 |
28.3 |
32.7 |
1444.3 |
Strain |
% S-9 |
Dose level of test item (µg/plate) |
PC |
|||||
0 |
50 |
150 |
500 |
1500 |
5000 |
|||
TA1535 |
10 |
10.0 |
14.0 |
10.3 |
9.3 |
13.7 |
14.7 |
126.0 |
TA1537 |
10 |
11.7 |
13.3 |
10.7 |
8.7 |
5.0 |
5.0 |
176.7 |
TA98 |
10 |
23.7 |
29.7 |
31.7 |
25.7 |
26.0 |
29.3 |
1063.0 |
TA100 |
10 |
81.3 |
94.7 |
103.3 |
75.7 |
66.3 |
68.0 |
1536.0 |
WP2uvrA |
10 |
31.0 |
35.7 |
29.7 |
30.7 |
36.0 |
39.0 |
180.7 |
PC: |
positive control |
|||||||
ppt: |
compound precipitation |
Applicant's summary and conclusion
- Conclusions:
- There were no biologically relevant or statistically significant increases in revertant numbers observed in any strain at any dose of Fenugreek Absolute, in the presence or absence of S 9 mix, under plate incorporation or pre-incubation conditions.
Fenugreek Absolute showed no mutagenic potential in the Bacterial Reverse Mutation Assay under the conditions of this test. - Executive summary:
Study Objective: The purpose of the bacterial reverse mutation test was to assess the mutagenic potential of the test item by its ability to induce reverse mutation in the specified bacterial strains from auxotrophic growth to prototrophy. Fenugreek Absolute was tested in vitro, for its ability to induce mutations in 4 histidine dependent auxotrophic mutants of Salmonella typhimurium, strains TA1535, TA1537, TA98 and TA100 and 1 tryptophan dependent auxotrophic mutant of Escherichia coli, WP2 uvrA.
Study Design: A range-finder experiment was performed to determine the cytotoxic range of the test item under plate incorporation conditions in TA98 and WP2 uvrA, in the presence and absence of S 9 mix (a liver post mitochondrial fraction derived from the livers of Aroclor 1254 treated rats). Mutation tests were then performed, using the plate incorporation method and pre-incubation method, in both the presence and absence of S 9 mix. The bacteria were exposed to the test item dissolved in dimethylsulphoxide (DMSO); the latter was also the negative Control. The positive Control chemicals were sodium azide (TA1535 and TA100), 9 aminoacridine (TA1537), 2 nitrofluorene (TA98) and 4 nitroquinoline-N-oxide (WP2 uvrA) in the absence of S 9 mix and 2-aminoanthracene (2AA) in the presence of S 9 mix (all strains).
The doses of Fenugreek Absolute used in Experiment 1 under plate incorporation conditions and Experiment 2 under pre-incubation conditions were 50, 150, 500, 1500 and 5000 µg/plate in all strains in the presence and absence of S 9 mix.
Results: All negative Controls gave mean counts of spontaneous revertant colonies within expected historical negative Control ranges obtained in this laboratory. All positive Controls induced marked increases in the number of revertant colonies and gave mean counts of induced revertant colonies which were within historical positive Control ranges. The responses to the positive Controls demonstrates that the bacteria were sensitive to the mutagens and that the S 9 mix was able to metabolise the pro-mutagen, 2AA, to a mutagen.
In Experiments 1 and 2, Fenugreek Absolute was tested up to the regulatory maximum dose level (5000 µg/plate) in all strains in the presence and absence of S 9 mix.
Precipitation was noted at 5000 µg/plate in all strains in the presence and absence of S-9 mix in both Experiments.
There were no biologically relevant or statistically significant increases in revertant numbers observed in any strain at any dose of Fenugreek Absolute, in the presence or absence of S 9 mix, under plate incorporation or pre-incubation conditions.
Conclusion: Fenugreek Absolute showed no mutagenic potential in the Bacterial Reverse Mutation Assay under the conditions of this test.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.