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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 282-004-4 | CAS number: 84082-58-6 Extractives and their physically modified derivatives such as tinctures, concretes, absolutes, essential oils, oleoresins, terpenes, terpene-free fractions, distillates, residues, etc., obtained from Origanum majorana, Labiatae.
- 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:
- 27 September to 16 October, 2017
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Remarks:
- GLP study conducted according to OECD test Guideline No. 471 without any deviation.
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 017
- Report date:
- 2017
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
- Version / remarks:
- 2011
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other:
- Version / remarks:
- ENV/EPOC(97)4 OECD Guideline for the Testing of Chemicals, Proposal for Replacement of Guidelines 471 and 472 Bacterial Reverse Mutation Test
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other:
- Version / remarks:
- EEC Directive 2000/32, L1362000, Annex 4D, dated May 19, 2000
- Deviations:
- no
- Principles of method if other than guideline:
- Not applicable
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Chemikaliengesetz (Chemicals Act) of the Federal Republic of Germany (inspected on July 13-16, 2015 / Signed on September 14, 2015)
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- Marjoram, sweet, ext.
- EC Number:
- 282-004-4
- EC Name:
- Marjoram, sweet, ext.
- Cas Number:
- 84082-58-6
- Molecular formula:
- Not relevant for a UVCB substance.
- IUPAC Name:
- 1-methyl-4-(propan-2-yl)cyclohexa-1,4-diene; 4-methyl-1-(propan-2-yl)cyclohex-3-en-1-ol
- Test material form:
- liquid
- Details on test material:
- - appearance: faint yellow liquid
- odor: spicy aromatic
- Batch No.of test material: 36024
- manufacturing date: 06/2016
- Expiration date of the lot/batch: June 2019
- N° CAS TSCA: 8015-01-8
- N° CAS EINECS: 84082-58-6
- Storage condition of test material: In the refrigerator (at +2 - +8° C), under nitrogen
Constituent 1
- Specific details on test material used for the study:
- STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Stability under test conditions: All formulations were prepared freshly before treatment and used within two hours of preparation.
- Solubility and stability of the test substance in the solvent/vehicle: Miscible in alcohol and other oils. The formulation was assumed to be stable for this period unless specified otherwise by the Sponsor.
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: Not reported
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):
- Not applicable
- Additional strain / cell type characteristics:
- not applicable
- Metabolic activation:
- with and without
- Metabolic activation system:
- 10% S9: S9-mix from the livers of male Wistar rats treated with Phenobarbital/ß-naphthoflavone (80 mg/kg bw/day by oral route).
- Test concentrations with justification for top dose:
- Experiment I (plate incorporation test):
In the pre-experiment, eight concentrations of the test item between 3 and 5000 μg/plate were tested as followed:
- All strains: 3; 10; 33; 100; 333; 1000; 2500; and 5000 μg/plate.
The pre-experiment is reported as experiment I.
Since toxic effects were observed a minimum of seven concentrations were tested in experiment II. 5000 μg/plate were chosen as maximal concentration.
Experiment II (pre-incubation test):
- Strain TA 100: 3; 10; 33; 100; 333; 1000; 2500; and 5000 μg/plate
- The remaining strains: 10; 33; 100; 333; 1000; 2500; and 5000 μg/plate - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used for test item preparation: Ethanol (purity 99.99%).
- Justification for choice of solvent: The solvent was chosen because of its solubility properties and its relative nontoxicity to the bacteria.
- Preparation of test formulation: All formulations were prepared freshly before treatment and used within two hours of preparation. The formulation was assumed to be stable for this period unless specified otherwise by the Sponsor.
Controlsopen allclose all
- Untreated negative controls:
- yes
- Remarks:
- untreated
- Negative solvent / vehicle controls:
- yes
- Remarks:
- ethanol
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- methylmethanesulfonate
- other: 4-nitro-o-phenylene-diamine
- Remarks:
- Without S9-mix
- Untreated negative controls:
- yes
- Remarks:
- untreated
- Negative solvent / vehicle controls:
- yes
- Remarks:
- ethanol
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene
- Remarks:
- With S9-mix
- Details on test system and experimental conditions:
- TEST SYSTEM:
The bacterial strains TA 1535, TA 1537, TA 98, TA 100, and WP2 uvrA were obtained from Trinova Biochem GmbH (35394 Gießen, Germany).
The histidine dependent strains are derived from Salmonella typhimurium strain LT2 through mutations in the histidine locus. Additionally due to the "deep rough" (rfa-minus) mutation they possess a faulty lipopolysaccharide envelope which enables substances to penetrate the cell wall more easily. A further mutation causes a reduction in the activity of an excision repair system. The latter alteration includes mutational processes in the nitrate reductase and biotin genes produced in a UV-sensitive area of the gene named "uvrB-minus". In the strains TA 98 and TA 100 the R-factor plasmid pKM 101 carries the ampicillin resistance marker.
Strain Escherichia coli WP2 and its derivatives carry the same defect in one of the genes for tryptophan biosynthesis. Tryptophan-independent (Trp+) mutants (revertants) can arise either by a base change at the site of the original alteration or by a base change elsewhere in the chromosome so that the original defect is suppressed. This second possibility can occur in several different ways so that the system seems capable of detecting all types of mutagen which substitute one base for another. Additionally, the uvrA derivative is deficient in the DNA repair process (excision repair damage). Such a repair-deficient strain may be more readily mutated by agents.
METHOD OF APPLICATION:
Experiment I: in agar (plate incorporation)
Experiment II: pre-incubation
DURATION
- Preincubation period: at 37°C for 60 minutes
- Exposure duration: plates were incubated upside down for at least 48 hours at 37°C
SELECTION AGENT (mutation assays): Plates with selective agar (without histidine/tryptophan) were used.
NUMBER OF REPLICATIONS: Triplicate plates per dose level.
DETERMINATION OF CYTOTOXICITY
- Method: Toxicity of the test item can be evident as a reduction in the number of spontaneous revertants or a clearing of the bacterial background lawn.
OTHERS:
- The colonies were counted using a validated computer system (cf. 3.8, Major computerized systems), which was connected to a PC with printer to print out the individual values, the means from the plates for each concentration together with standard deviations and enhancement factors as compared to the spontaneous reversion rates. Due to precipitation of the test item the colonies were partly counted manually. - Evaluation criteria:
- A test item is considered as a mutagen if a biologically relevant increase in the number of revertants exceeding the threshold of twice (strains TA 98, TA 100, and WP2 uvrA) or thrice (strains TA 1535 and TA 1537) the colony count of the corresponding solvent control is observed.
If exposure to a test item does not produce a reproducible increase in mean revertant colony numbers, it will be considered to show no evidence of mutagenic activity in this test system. No statistical analysis will be performed.
A dose dependent increase is considered biologically relevant if the threshold is exceeded at more than one concentration.
An increase exceeding the threshold at only one concentration is judged as biologically relevant if reproduced in an independent second experiment.
A dose dependent increase in the number of revertant colonies below the threshold is regarded as an indication of a mutagenic potential if reproduced in an independent second experiment. However, whenever the colony counts remain within the historical range of negative and solvent controls such an increase is not considered biologically relevant.
If the results obtained fail to satisfy the criteria for a clear “positive” or “negative” response, even after additional testing, it is acceptable to conclude an equivocal response. - Statistics:
- No statistical analysis was performed as no evidence of mutagenic activity was shown in this test system.
Results and discussion
Test resultsopen allclose all
- Key result
- Species / strain:
- S. typhimurium, other: TA 1535, TA 1537, TA 98 and TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- 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:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- 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 in the test tubes in experiment I from 1000 to 5000 μg/plate with S9 mix and in experiment II at 5000 μg/plate without S9 mix and from 1000 to 5000 μg/plate with S9 mix. Precipitation of the test item in the overlay agar on the incubated agar plates was observed in experiment I at 5000 μg/plate without S9 mix and from 1000 to 5000 μg/plate with S9 mix and in experiment II from 2500 to 5000 μg/plate with and without S9 mix. The undissolved particles had no influence on the data recording.
- Other confounding effects: Note reported
HISTORICAL CONTROL DATA: The colony counts remain within the historical range of negative and solvent controls.
The laboratory´s historical control data from November 2014 until November 2016 represent approx. 600 experiments (WP2 uvrA the historical data are based on approx. 350 experiments).
- Positive historical control data as Mean value of revertants/plate (SD):
TA 1535: 1130 (143.1) -S9-mix ; 388 (58.2) +S9-mix
TA 1537: 82 (12.7) -S9-mix ; 191 (60.8) +S9-mix
TA 98: 378 (73.7) -S9-mix ; 3949 (771.8) +S9-mix
TA 100: 1966 (293.2) -S9-mix ; 3798 (830.4) +S9-mix
WP2 uvrA: 798 (362.7) -S9-mix ; 378 (112.6) +S9-mix
- Negative (solvent) historical control data as Mean value of revertants/plate (SD):
TA 1535: 12 (2.5) -S9-mix ; 12 (2.5) +S9-mix
TA 1537: 10 (2.2) -S9-mix ; 13 (3.5) +S9-mix
TA 98: 25 (4.4) -S9-mix ; 34 (6.2) +S9-mix
TA 100: 156 (26.0) -S9-mix ; 148 (32.3) +S9-mix
WP2 uvrA: 41 (5.6) -S9-mix ; 50 (6.8) +S9-mix
- Negative (untreated) historical control data as Mean value of revertants/plate (SD):
TA 1535: 12 (3.1) -S9-mix ; 12 (2.9) +S9-mix
TA 1537: 11 (2.7) -S9-mix ; 14 (4.0) +S9-mix
TA 98: 27 (4.9) -S9-mix ; 37 (6.5) +S9-mix
TA 100: 176 (23.6) -S9-mix ; 172 (25.4) +S9-mix
WP2 uvrA: 42 (5.8) -S9-mix ; 52 (6.8) +S9-mix
ADDITIONAL INFORMATION ON CYTOTOXICITY
See 'Any other information on results incl. tables'
MUTAGENICITY RESULTS
See 'Any other information on results incl. tables'
Any other information on results incl. tables
Table 7.6.1/2: Reduced background growth observed at the following concentrations (µg/plate):
Strain |
Experiment I |
Experiment II |
||
|
Without S9-mix |
With S9-mix |
Without S9-mix |
With S9-mix |
TA 1535 |
/ |
/ |
2500 – 5000 |
2500 – 5000 |
TA 1537 |
/ |
/ |
2500 – 5000 |
1000 – 5000 |
TA 98 |
/ |
/ |
1000 – 5000 |
1000 – 5000 |
TA 100 |
/ |
/ |
1000 – 5000 |
333 – 5000 |
WP2 uvrA |
/ |
/ |
2500 – 5000 |
2500 – 5000 |
/ = normal background growth
Table 7.6.1/3: Reduction in the number of revertants (below the induction factor of 0.5) observed at the following concentrations (µg/plate):
Strain |
Experiment I |
Experiment II |
||
|
Without S9-mix |
With S9-mix |
Without S9-mix |
With S9-mix |
TA 1535 |
2500 – 5000 |
/ |
2500 – 5000 |
2500 – 5000 |
TA 1537 |
2500 – 5000 |
2500 – 5000 |
2500 – 5000 |
1000 – 5000 |
TA 98 |
5000 |
5000 |
1000 – 5000 |
2500 – 5000 |
TA 100 |
1000 – 5000 |
1000 – 5000 |
333 - 5000 |
333 – 5000 |
WP2 uvrA |
/ |
2500 – 5000 |
2500 – 5000 |
2500 – 5000 |
/ = no toxic effects, evident as a reduction in the number of revertants (below the induction factor of 0.5)
Applicant's summary and conclusion
- Conclusions:
- Under the test condition, test item is not mutagenic with and without metabolic activation in S.typhimurium (strains TA1535, TA1537, TA98 and TA100) and E.coli WP2 uvrA according to the criteria of the Annex VI of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS
- Executive summary:
In a reverse gene mutation assay performed according to the OECD test guideline No. 471 and in compliance with GLP, Salmonella typhimurium strains TA1535, TA1537, TA98 and TA100 and Escherichia coli strain WP2 uvrA were treated with the test item using both the Ames plate incorporation and pre-incubation methods at up to eight dose levels, in triplicate, both with and without the addition of a rat liver homogenate metabolizing system (10% liver S9 in standard co-factors). The dose range for the range-finding test (plate incorporation, Experiment I) was predetermined as 3 to 5000 μg/plate. Based on the results of the experiment I, the experiment II was performed using the pre-incubation method at concentrations from 3 to 5000 µg/plate, depending on the bacterial strain type and presence or absence of S9 -mix. Negative, solvent (ethanol) and positive control groups were also included in mutagenicity tests.
The test item precipitated in the overlay agar in the test tubes in experiment I from 1000 to 5000 μg/plate with S9 mix and in experiment II at 5000 μg/plate without S9 mix and from 1000 to 5000 μg/plate with S9 mix. Precipitation of the test item in the overlay agar on the incubated agar plates was observed in experiment I at 5000 μg/plate without S9 mix and from 1000 to 5000 μg/plate with S9 mix and in experiment II from 2500 to 5000 μg/plate with and without S9 mix. The undissolved particles had no influence on the data recording.
The plates incubated with the test item showed reduced background growth in experiment II in all strains with and without S9 mix. In experiment I normal background growth was observed up to 5000 μg/plate with and without S9 mix in all strains used.
Toxic effects, evident as a reduction in the number of revertants (below the indication factor of 0.5), occurred in all strains with and without metabolic activation.
No substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with test material at any dose level, neither in the presence nor absence of metabolic activation (S9 mix). There was also no tendency of higher mutation rates with increasing concentrations in the range below the generally acknowledged border of biological relevance.
Appropriate reference mutagens were used as positive controls and showed a distinct increase of induced revertant colonies.
In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used.
Under the test condition, test item is not mutagenic with and without metabolic activation in S. typhimurium (strains TA1535, TA1537, TA98 and TA100) and E. coli WP2 uvrA according to the criteria of the Annex VI of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.
This study is considered as acceptable and satisfies the requirement for reverse gene mutation endpoint.
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.