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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 237-301-3 | CAS number: 13732-62-2
- 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 mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- migrated information: read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: The study is scientifically valid and followed the principles of OECD Test Guideline 476.
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 979
- Report date:
- 1979
Materials and methods
Test guideline
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- not specified
- GLP compliance:
- no
- Type of assay:
- mammalian cell gene mutation assay
Test material
- Reference substance name:
- Morpholine
- EC Number:
- 203-815-1
- EC Name:
- Morpholine
- Cas Number:
- 110-91-8
- Molecular formula:
- C4H9NO
- IUPAC Name:
- morpholine
- Details on test material:
- - Name of test material: Morpholine (7H-4892/LOS-0575)
- Physical state: clear colorless liquid
Constituent 1
Method
- Target gene:
- thymidine kinase locus
Species / strain
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- - Type and identity of media: The cells were maintained in Fischer's mouse leukemia medium supplemented with L-glutamine, sodium pyruvate, and horse serum (10 % by volume). Cloning medium consisted of the preceding growth medium with the addition of agar to a final concentration of 0.35 % to achieve a semisolid state. Section medium was cloning medium containing 100 µg/mL of BrdU or 3 µg/mL of TFT.
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: no
- Periodically "cleansed" against high spontaneous background: yes - Additional strain / cell type characteristics:
- other: heterozygous at the thymidine kinase locus
- Metabolic activation:
- with and without
- Metabolic activation system:
- Aroclor 1254 induced rat liver S9 (S9 mix)
- Test concentrations with justification for top dose:
- Without metabolic activation:
Experiment 1: 0.078, 0.500, 0.625, 1.000, 1.250 µL/mL
Experiment 2: 0.500, 0.625, 0.750, 1.000, 1.250 µL/mL
Experiment 3: 0.750, 1.000, 1.200 µL/mL
Metabolic activation:
Experiment 1: 0.156, 0.500, 0.625, 1.000, 1.250 µL/mL
Experiment 2: 0.500, 0.625, 0.750, 1.000, 1.250 µL/mL
Experiment 3: 0.750, 1.000 µL/mL - Vehicle / solvent:
- - Solvent used: DMSO
- Justification for choice of solvent/vehicle: due to a technical error in the initial solubility testing, the test substance was described as insoluble at 100 uL/mL in deionized water. The substance was, in fact, soluble and water would have been the preferred solvent.
Controlsopen allclose all
- Untreated negative controls:
- yes
- Remarks:
- untreated
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- no
- Positive controls:
- yes
- Remarks:
- see positive controls below
- Remarks:
- with and without metabolic activation
- Untreated negative controls:
- yes
- Remarks:
- untreated
- Negative solvent / vehicle controls:
- not specified
- Remarks:
- no data was provided on the solvent which the positive control was dissolved in.
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- ethylmethanesulphonate
- Remarks:
- without metabolic activation
Migrated to IUCLID6: 0.5 µL/mL
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- not specified
- Remarks:
- No data was provided on the solvent which the positive control was dissolved in.
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: dimethylnitrosamine (0.3 µL/mL)
- Remarks:
- with metabolic activation
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 4 hours
- Expression time (cells in growth medium): 2-3 days
- Selection time (if incubation with a selection agent): 10 days
- Fixation time (start of exposure up to fixation or harvest of cells): cells were harvested at the end of the selection time.
SELECTION AGENT (mutation assays): 5-bromo-2'deoxyuridine or 5-trifluorothymidine
NUMBER OF REPLICATIONS: 1-2
NUMBER OF CELLS EVALUATED: Mutation frequency was based on 10 EXP 6 cells.
DETERMINATION OF CYTOTOXICITY
- Method: % relative total growth - Evaluation criteria:
- The test substance was considered mutagenic in the assay if:
- A dose response relationship was observed over 3 of the 5 dose levels employed.
- The minimum increase at the low level of the dose response curve was at least 2.5 times greater than the solvent and/or negative control values.
- The solvent and negative control data were within the normal range of the spontaneous background for the TK locus.
All evaluations of mutagenic activity were based on consideration of the concurrent solvent and negative control values run with the experiment in question. Positive control values were not used as reference points, but were included to ensure that the current cell population responded to direct and promutagens under the appropriate treatment conditions.
Occasionally, a single point within a concentration range was expected to show an increase 2.5 times greater than the spontaneous background. If the increase was at the high dose, was reproducible, and if an additional higher dose level was not feasible because of toxicity, the substance was possibly considered mutatgenic. If the increase was internal within the dose range and was not reproducible, the increase was considered aberrant. If the internal increase was reproducible, several doses clustered around the positive concentration were examined to either confirm or reject the reliability of the effect. - Statistics:
- Not indicated.
Results and discussion
Test resultsopen allclose all
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- without
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with
- 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:
- RANGE-FINDING/SCREENING STUDIES: The test substance was tested for cytotoxicity at an applied dose range of 0.01-5 uL/mL. After an exposure time of four hours, the cells were washed and a viable cell count was obtained the next day. Relative cytotoxicities expressed as the reduction in growth compared to the growth of untreated cells were used to select seven to ten doses that covered the range from 0 to 50-90 % reduction in 24 hour growth. These selected doses were subsequently applied to cell cultures prepared for mutagenicity testing, but only four or five of the doses would be carried through the mutant selection process. The test substance remained soluble in the growth medium, but an alkaline pH was obtained at concentrations above about 0.3 µL/mL. Twenty four hours after treatment, the cell count was greatly reduced at 1.25 µL/mL and doses of 2.5 µL/mL and higher were completely lethal.
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
Any other information on results incl. tables
Four trials of the mutation assay were initiated, but the first trial became contaminated before any mutagenesis data could be collected. Under the nonactivation conditions, the test substance was lethal at 2.0 µL/mL and five treatments from 1.25 -0.078 µL/mL were assayed for mutant induction. The 1.25 µL/mL treatment was highly toxic (9.4 % relative growth) and did not induce a significant increase in mutant frequency. A 2.5 fold increase over the background was considered the minimum criterion for demonstrating mutagenesis at any given dose level. The other four treatments were moderately toxic and also yielded mutant frequencies similar to the background, except for the 1 µL/mL treatment. In this case, a 2.8 fold increase was observed. An increase of this small magnitude that is not followed by a similar or larger increase for more toxic treatments is normally regarded as spurious. Nevertheless, a repeat assay was performed to test the repeatability of the response. In the second nonactivation assay, the toxicity of the test substance was more variable as a function of the applied concentration. Three of the assayed treatments were highly toxic and all of these induced mutant frequency increases near the 2.5 fold criterion used for evaluation as mutagenic. Thus, 2.2 fold, 2.4 fold and 2.7 fold increases were obtained for 0.625, 1 and 1.25 µL/mL, respectively. These results suggested that the test substance may be weakly mutagenic at the limit of detectability for this assay. A third nonactivation assay was performed which yielded similar behavior. One treatment with 0.75 µL/mL and another at 1.2 µL/mL induced 2.5 and 2.7 fold increases in the mutant frequency. However, duplicate treatments at these two doses, both of which were highly toxic, caused no significant change from the background frequency. The results from all three trials therefore indicated that the test substance can be weakly mutagenic for highly toxic treatments in the 0.625 to 1.25 µL/mL dose range. The lack of mutagenic activity for some toxic treatments in this dose range suggested competition between toxic and mutagenic modes of action. In the presence of metabolic activation, the test substance was not as toxic, indicating a reaction with the activating system. Only moderately toxic treatments were achieved in the first trial for applied concentrations up to 1.25 µL/mL (35.5 % relative growth). The mutant frequencies in the treated cultures remained similar to the background. The next highest applied concentration of 2 µL/mL was completely lethal to the cells. A repeat assay gave similar results and confirmed the lack of detectable mutagenesis for moderately toxic treatments obtained with concentrations up to 1.25 µL/mL. In this assay, the next highest dose was 1.5 µL/mL, which was completely lethal. A third activation assay was performed in which most of the applied concentrations (from 1.2 to 2 µL/mL) were lethal and only two dose levels (in duplicate) were available for analysis. One of the 0.75 µL/mL treatments was moderately toxic (37.3 % relative growth), whereas the other treatments had no apparent effect on the cells. The mutant frequencies remained similar to the one solvent negative control that was not contaminated. Thus, the test substance had little to moderate toxicity in the presence of activation until a threshold concentration was reached where lethality suddenly became excessive. This threshold occurred between 1.25 and 1.5 µL/mL in trial 2 and between 1.0 and 1.2 µL/mL in trial 3. Since the test substance was alkaline in the culture medium at these concentrations, the sharp increase in lethality may be pH related rather than test substance induced. Any mutagenic activity that might be associated with highly toxic treatments would therefore occur over a very narrow concentration range just preceding complete lethality. Such information was probably not biologically meaningful, so further attempts to achieve this toxicity range did not seem warranted. The average cloning efficiencies for the solvent and untreated negative control varied from 105 to 108% without activation and from 106 to 124 % with activation, which demonstrated excellent culturing conditions for the assays. The negative control mutant frequencies were all in the normal range, and the positive control substance yielded normal mutant frequencies that were greatly in excess of the background.
This study is classified as acceptable. This study satisfies the requirement for Test Guideline OECD .
Applicant's summary and conclusion
- Conclusions:
- Interpretation of results (migrated information):
negative with metabolic activation
positive without metabolic activation
Morpholine was evaluated for mutagenic potential in the mouse lymphoma forward mutation assay using L5178Y TK +/- cells in the presence and absence of metabolic activation (S9 mix). Under the conditions of this study, the test substance was considered to be weakly mutagenic in the assay without metabolic activation.
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.