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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 294-585-1 | CAS number: 91744-23-9
- 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
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
The substance was negative in a reliable in vitro bacterial Reverse Mutation Assay conducted according to OECD 471, employing Salmonella typhimurium strains TA 1535, TA 1537, TA 98 and TA 100 exposed to the substance at concentrations up to 5000 µg/plate (plate incorporation method) in the presence and absence of mammalian metabolic activation (Aroclor 1254 induced liver S9 mix). Structural analogues of the substance were also found to be negative in an in vitro bacterial gene mutation assay and in an in vitro mammalian cell point mutation assay. The in vitro cytogenetics assay has been waived on the basis that a reliable in vivo study is available on a structural analogue.
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Study performed according to internationally accepted testing guideline, well documented.
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- not specified
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Additional strain / cell type characteristics:
- other: TA 98 his mutation: D3052; other markers: rfa uvrB; plasmid: pKM101 / TA 100 his mutation: G46; other markers: rfa uvrB; plasmid: pKM101 / TA 1535 his mutation: G46; other markers: rfa uvrB; / TA 1537 his mutation: C3076; other markers: rfa uvrB.
- Metabolic activation:
- with and without
- Metabolic activation system:
- Aroclor 1254 induced liver S9 mix
- Test concentrations with justification for top dose:
- 50 / 160 / 500 / 1600 / 5000 µg/plate
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: tetrahydrofurane
- Justification for choice of solvent/vehicle: not mentioned - Untreated negative controls:
- yes
- Remarks:
- water
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO and tetrahydrofurane
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-Nitrofluorene (2.5 µg/plate) for the strains TA 98; Sodium azide (5.0 µg/plate) for TA 100; Sodium azide (2.5 µg/plate) for TA 1535; 9-Aminoacridine (5.0 µg/plate) for TA 1537.
- Remarks:
- without metabolic activation
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-Aminoanthracene (2.5 µg/plate) with all strains
- Remarks:
- with metabolic activation
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation) and preincubation, performed in two independent tests
DURATION
- Preincubation period: 30 minutes
- Exposure duration: 96 hours
NUMBER OF REPLICATIONS: 3 per concentration
DETERMINATION OF CYTOTOXICITY
- Method: not mentioned
OTHER EXAMINATIONS:
- Other: Determination of the frequency of induced or spontaneous reversion to histidine independence with negative controls (H2O), solvent controls (DMSO), test substance concentrations and positive controls; determination of the titers of overnight cultures
OTHER: none - Evaluation criteria:
- According to Ames a test article which caused no mutagenic effects at a concentration of 5000 µg/plate will be called non-mutagenic.
- Statistics:
- no statistics performed
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no 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 measured
- Effects of osmolality: not applicable
- Evaporation from medium: not applicable
- Water solubility: not mentioned
- Precipitation: no
- Other confounding effects: none
RANGE-FINDING/SCREENING STUDIES: not performed
COMPARISON WITH HISTORICAL CONTROL DATA: not performed
ADDITIONAL INFORMATION ON CYTOTOXICITY: none - Remarks on result:
- other: strain/cell type: Salmonella typhimurium
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- The test substance is negative in the Bacterial Reverse Mutation Assay with and without metabolic activation.
- Executive summary:
In a Bacterial Reverse Mutation Assay according to OECD 471, strains TA 1535, TA 1537, TA 98 and TA 100 of S. typhimurium were exposed to the test substance (CAS 91744-23-9) in solvent tetrahydrofurane at concentrations of 50 / 160 / 500 / 1600 / 5000 µg/plate (plate incorporation) in the presence and absence of mammalian metabolic activation (aroclor 1254 induced liver S9 mix).
There was no evidence of induced mutant colonies over background.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2012-06-06 until 2012-09-19
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- Principles of method if other than guideline:
- first experiment 4 hours treatment with and without metabolic activation
second experiment 24 hours treatment without metabolic activation, 4 hours treatment with metabolic activation - GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- HPRT
- Species / strain / cell type:
- Chinese hamster lung fibroblasts (V79)
- Details on mammalian cell type (if applicable):
- - Type and identity of media: MEM
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: yes
- Periodically "cleansed" against high spontaneous background: yes - Metabolic activation:
- with and without
- Metabolic activation system:
- Phenobarbital/Beta-Naphtoflavone induced Rat liver S9
- Test concentrations with justification for top dose:
- Experiment I:
without metabolic activation: 4.7; 9.4; 18.8; 37.5; 75.0; 150.0 µg/mL
with metabolic activation: 4.7; 9.4; 18.8; 37.5; 75.0; 150.0 µg/mL
Experiment II:
without metabolic activation: 4.7; 9.4; 18.8; 37.5; 75.0; 150.0 µg/mL
with metabolic activation: 4.7; 9.4; 18.8; 37.5; 75.0; 150.0 µg/mL
The cultures at the lowest concentration of 4.7 µg/mL in the presence and absence of metabolic activation (experiment I and II) were not continued since a minimum of only four analysable concentrations is required by the guidelines. - Vehicle / solvent:
- - Solvent used: Tetrahydrofuran (THF) (99.9%)
- Justification for choice of solvent/vehicle: Solubility properties - Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- ethylmethanesulphonate
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 7,12-dimethylbenzanthracene
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: Experiment I: 4 hours with and without metabolic activation, Experiment II: 24 hours without metabolic activation, 4 hours with metabolic activation
- Expression time (cells in growth medium): 72 hours
- Selection time (if incubation with a selection agent): 10 days
SELECTION AGENT (mutation assays): 6-Thioguanine
NUMBER OF REPLICATIONS: 2
NUMBER OF CELLS EVALUATED: >1,5x10exp. 6
DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency - Evaluation criteria:
- A test item producing neither a concentration-related increase of the mutant frequency nor a reproducible positive response at any of the test points is considered to be non-mutagenic in this system.
A mutagenic response is described as follows:
The test item is classified as mutagenic if it induces reproducibly with one of the concen¬trations a mutation frequency that is three times higher than the spontaneous mutation fre¬quency in the experiment.
The test item is classified as mutagenic if there is a reproducible concentration-related increase of the mutation frequency. Such evaluation may be considered also in the case that a threefold increase of the mutant frequency is not observed.
In a case by case evaluation this decision depends on the level of the correspon¬ding solvent control data. - Statistics:
- A linear regression (least squares) was performed to assess a possible dose dependent increase of mutant frequencies. The number of mutant colonies obtained for the groups treated with the test item were compared to the solvent control groups. A trend is judged as significant whenever the p-value (probability value) is below 0.05. However, both, biological and statistical significance were considered together.
- Species / strain:
- Chinese hamster lung fibroblasts (V79)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: Not effected pH 7.32 in the solvent control versus pH 7.31 at 1200 µg test item/mL
- Effects of osmolality: Not increased (357 mOsm in the solvent control versus 342 mOsm at 1200 µg test item/mL
- Evaporation from medium: Not examined
- Water solubility: Not indicated
- Precipitation:
In the first experiment precipitation of the test item at the end of treatment was noted at 75 and 150 µg/mL with and without metabolic activation. In the second experiment precipitation as described above occurred at 75 and 150 µg/mL with and at 37.5 µg/mL and above without metabolic activation.
- Other confounding effects: None
RANGE-FINDING/SCREENING STUDIES:
The highest concentration used in the pre-test was 1200 µg/mL limited by the solubility of the test item in THF and aqueous medium. Test item concentrations between 9.4 µg/mL and 1200 µg/mL were used to evaluate toxicity in the presence (4 hours treatment) and absence (4 hours and 24 hours treatment) of metabolic activation. Relevant cytotoxic effects indicated by a relative suspension growth below 50 were noted at 1200 µg/mL following 24 hours treatment without metabolic activation. The test medium was checked for precipitation or phase separation at the end of each treatment period (4 or 24 hours) prior to removal to the test item.
The test medium was checked for precipitation or phase separation at the end of each treatment period (4 or 24 hours) prior to removal to the test item. Precipitation occurred at 75.0 µg/mL and above in the presence (4 hours treatment) and absence (4 and 24 hours treatment) of metabolic activation. .
COMPARISON WITH HISTORICAL CONTROL DATA: Complies
ADDITIONAL INFORMATION ON CYTOTOXICITY:
No relevant toxic effects occurred up to the maximum concentration with and without metabolic activation following 4 and 24 hours treatment. - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results (migrated information):
negative
The test item did not induce gene mutations at the HPRT locus in V79 cells. Therefore, Glycerides, C16-18 mono-, di and tri-, hydrogenated, citrates, potassium salts is considered to be non-mutagenic in this HPRT assay. - Executive summary:
The test item (Glycerides, C16-18 mono-, di and tri-, hydrogenated, citrates, potassium salts) was assessed for its potential to induce gene mutations at the HPRT locus using V79 cells of the Chinese hamster.
The study was performed in two independent experiments, using identical experimental procedures. In the first experiment the treatment period was 4 hours with and without metabolic activation. The second experiment was performed with a treatment time of 4 hours with and 24 hours without metabolic activation.
The maximum concentration was limited by the solubility of the test item in THF and aqueous medium. Precipitation of the test item at the end of treatment was noted at 75 and 150 µg/mL in the first experiment with and without metabolic activation. In the second experiment precipitation as described above occurred at 75 and 150 µg/mL with and at 37.5 µg/mL and above without metabolic activation.
No relevant toxic effects occurred up to the maximum concentration with and without metabolic activation following 4 and 24 hours treatment.
No relevant and reproducible increase in mutant colony numbers/106cells was observed in the main experiments up to the maximum concentration. The induction factor exceeded the threshold of three times the corresponding solvent control and the range of the historical solvent control data in the second culture of the first experiment without metabolic activation at 18.8 and 150.0 µg/mL. However, the increase at 18.8 µg/mL was marginal (induction factor of 3.4) and was not reproduced in the parallel culture under identical conditions. The increase at 150 µg/mL was substantial (induction factor of 6.1) but again, not reproduced in the parallel culture under identical experimental conditions. Furthermore, the concentration of 150 µg/mL was the second precipitating concentration so, the irreproducible increase of the mutation frequency was judged as irrelevant precipitation artefact. The minor increase at 18.8 µg/mL was judged as biologically irrelevant fluctuation.
A linear regression analysis (least squares) was performed to assess a possible dose dependent increase of mutant frequencies. A significant dose dependent trend of the mutation frequency indicated by a probability value of <0.05 was detected in both cultures of the first experiment without metabolic activation. The trend observed in culture I however, was judged as irrelevant as it actually was reciprocal, going down versus increasing concentrations. The trend observed in culture II was judged irrelevant as it was based on a precipitation artefact at 150 µg/mL as described above.
In both experiments of this study (with and without S9 mix) the range of the solvent controls was from 9.7 up to 35.2 mutants per 106cells; the range of the groups treated with the test item was from 2.6 up to 84.9 mutants per 106cells.
EMS (150 µg/mL) and DMBA (1.1 µg/mL) were used as positive controls and showed a distinct increase in induced mutant colonies.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of 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:
- guideline study
- Remarks:
- Read-across to K1 study therefore K2 is the maximum Klimisch value that can be assigned
- Justification for type of information:
- Please see the read-across report attached in section 13 of this dossier.
- Reason / purpose for cross-reference:
- read-across source
- Target gene:
- HPRT
- Species / strain / cell type:
- Chinese hamster lung fibroblasts (V79)
- Details on mammalian cell type (if applicable):
- - Type and identity of media: MEM
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: yes
- Periodically "cleansed" against high spontaneous background: yes - Metabolic activation:
- with and without
- Metabolic activation system:
- Phenobarbital/Beta-Naphtoflavone induced Rat liver S9
- Test concentrations with justification for top dose:
- Experiment I:
without metabolic activation: 4.7; 9.4; 18.8; 37.5; 75.0; 150.0 µg/mL
with metabolic activation: 4.7; 9.4; 18.8; 37.5; 75.0; 150.0 µg/mL
Experiment II:
without metabolic activation: 4.7; 9.4; 18.8; 37.5; 75.0; 150.0 µg/mL
with metabolic activation: 4.7; 9.4; 18.8; 37.5; 75.0; 150.0 µg/mL
The cultures at the lowest concentration of 4.7 µg/mL in the presence and absence of metabolic activation (experiment I and II) were not continued since a minimum of only four analysable concentrations is required by the guidelines. - Vehicle / solvent:
- - Solvent used: Tetrahydrofuran (THF) (99.9%)
- Justification for choice of solvent/vehicle: Solubility properties - Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- ethylmethanesulphonate
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 7,12-dimethylbenzanthracene
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: Experiment I: 4 hours with and without metabolic activation, Experiment II: 24 hours without metabolic activation, 4 hours with metabolic activation
- Expression time (cells in growth medium): 72 hours
- Selection time (if incubation with a selection agent): 10 days
SELECTION AGENT (mutation assays): 6-Thioguanine
NUMBER OF REPLICATIONS: 2
NUMBER OF CELLS EVALUATED: >1,5x10exp. 6
DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency - Evaluation criteria:
- A test item producing neither a concentration-related increase of the mutant frequency nor a reproducible positive response at any of the test points is considered to be non-mutagenic in this system.
A mutagenic response is described as follows:
The test item is classified as mutagenic if it induces reproducibly with one of the concen¬trations a mutation frequency that is three times higher than the spontaneous mutation fre¬quency in the experiment.
The test item is classified as mutagenic if there is a reproducible concentration-related increase of the mutation frequency. Such evaluation may be considered also in the case that a threefold increase of the mutant frequency is not observed.
In a case by case evaluation this decision depends on the level of the correspon¬ding solvent control data. - Statistics:
- A linear regression (least squares) was performed to assess a possible dose dependent increase of mutant frequencies. The number of mutant colonies obtained for the groups treated with the test item were compared to the solvent control groups. A trend is judged as significant whenever the p-value (probability value) is below 0.05. However, both, biological and statistical significance were considered together.
- Species / strain:
- Chinese hamster lung fibroblasts (V79)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: Not effected pH 7.32 in the solvent control versus pH 7.31 at 1200 µg test item/mL
- Effects of osmolality: Not increased (357 mOsm in the solvent control versus 342 mOsm at 1200 µg test item/mL
- Evaporation from medium: Not examined
- Water solubility: Not indicated
- Precipitation:
In the first experiment precipitation of the test item at the end of treatment was noted at 75 and 150 µg/mL with and without metabolic activation. In the second experiment precipitation as described above occurred at 75 and 150 µg/mL with and at 37.5 µg/mL and above without metabolic activation.
- Other confounding effects: None
RANGE-FINDING/SCREENING STUDIES:
The highest concentration used in the pre-test was 1200 µg/mL limited by the solubility of the test item in THF and aqueous medium. Test item concentrations between 9.4 µg/mL and 1200 µg/mL were used to evaluate toxicity in the presence (4 hours treatment) and absence (4 hours and 24 hours treatment) of metabolic activation. Relevant cytotoxic effects indicated by a relative suspension growth below 50 were noted at 1200 µg/mL following 24 hours treatment without metabolic activation. The test medium was checked for precipitation or phase separation at the end of each treatment period (4 or 24 hours) prior to removal to the test item.
The test medium was checked for precipitation or phase separation at the end of each treatment period (4 or 24 hours) prior to removal to the test item. Precipitation occurred at 75.0 µg/mL and above in the presence (4 hours treatment) and absence (4 and 24 hours treatment) of metabolic activation. .
COMPARISON WITH HISTORICAL CONTROL DATA: Complies
ADDITIONAL INFORMATION ON CYTOTOXICITY:
No relevant toxic effects occurred up to the maximum concentration with and without metabolic activation following 4 and 24 hours treatment. - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results (migrated information):
negative
The test item did not induce gene mutations at the HPRT locus in V79 cells. Therefore, Glycerides, C16-18 mono-, di and tri-, hydrogenated, citrates, potassium salts is considered to be non-mutagenic in this HPRT assay. - Executive summary:
The test item (Glycerides, C16-18 mono-, di and tri-, hydrogenated, citrates, potassium salts) was assessed for its potential to induce gene mutations at the HPRT locus using V79 cells of the Chinese hamster.
The study was performed in two independent experiments, using identical experimental procedures. In the first experiment the treatment period was 4 hours with and without metabolic activation. The second experiment was performed with a treatment time of 4 hours with and 24 hours without metabolic activation.
The maximum concentration was limited by the solubility of the test item in THF and aqueous medium. Precipitation of the test item at the end of treatment was noted at 75 and 150 µg/mL in the first experiment with and without metabolic activation. In the second experiment precipitation as described above occurred at 75 and 150 µg/mL with and at 37.5 µg/mL and above without metabolic activation.
No relevant toxic effects occurred up to the maximum concentration with and without metabolic activation following 4 and 24 hours treatment.
No relevant and reproducible increase in mutant colony numbers/106cells was observed in the main experiments up to the maximum concentration. The induction factor exceeded the threshold of three times the corresponding solvent control and the range of the historical solvent control data in the second culture of the first experiment without metabolic activation at 18.8 and 150.0 µg/mL. However, the increase at 18.8 µg/mL was marginal (induction factor of 3.4) and was not reproduced in the parallel culture under identical conditions. The increase at 150 µg/mL was substantial (induction factor of 6.1) but again, not reproduced in the parallel culture under identical experimental conditions. Furthermore, the concentration of 150 µg/mL was the second precipitating concentration so, the irreproducible increase of the mutation frequency was judged as irrelevant precipitation artefact. The minor increase at 18.8 µg/mL was judged as biologically irrelevant fluctuation.
A linear regression analysis (least squares) was performed to assess a possible dose dependent increase of mutant frequencies. A significant dose dependent trend of the mutation frequency indicated by a probability value of <0.05 was detected in both cultures of the first experiment without metabolic activation. The trend observed in culture I however, was judged as irrelevant as it actually was reciprocal, going down versus increasing concentrations. The trend observed in culture II was judged irrelevant as it was based on a precipitation artefact at 150 µg/mL as described above.
In both experiments of this study (with and without S9 mix) the range of the solvent controls was from 9.7 up to 35.2 mutants per 106cells; the range of the groups treated with the test item was from 2.6 up to 84.9 mutants per 106cells.
EMS (150 µg/mL) and DMBA (1.1 µg/mL) were used as positive controls and showed a distinct increase in induced mutant colonies.
Referenceopen allclose all
Table #1: Plate incorporation test: Number of revertants per plate (mean of 3 plates)
|
[Strain TA 98] |
[Strain TA 100] |
[Strain TA 1535] |
||||||
Conc. |
- MA |
+ MA |
Cytotoxic |
- MA |
+ MA |
Cytotoxic |
- MA |
+ MA |
Cytotoxic |
0* |
16 ± 3 |
48 ± 10 |
no |
146 ± 16 |
168 ± 10 |
no |
8 ± 3 |
12 ± 3 |
no |
50 |
14 ± 2 |
39 ± 8 |
no |
166 ± 1 |
163 ± 8 |
no |
6 ± 1 |
9 ± 2 |
no |
160 |
18 ± 1 |
50 ± 5 |
no |
136 ± 16 |
168 ± 3 |
no |
7 ± 1 |
15 ± 4 |
no |
500 |
15 ± 7 |
49 ± 8 |
no |
182 ± 13 |
172 ± 9 |
no |
10 ± 3 |
12 ± 4 |
no |
1600 |
18 ± 7 |
59 ± 9 |
no |
175 ± 11 |
163 ± 15 |
no |
7 ± 1 |
12 ± 5 |
no |
5000 |
21 ± 4 |
55 ± 4 |
no |
166 ± 18 |
179 ± 8 |
no |
7 ± 2 |
18 ± 4 |
no |
Positive control |
8.4 |
7.3 |
no |
499 ± 38 |
2028 ± 39 |
no |
242 ± 34 |
215 ± 14 |
no |
*solvent control with THF
Table #2: Plate incorporation test: Number of revertants per plate (mean of 3 plates)
|
[Strain TA 1537] |
||
Conc. |
- MA |
+ MA |
Cytotoxic |
0* |
7 ± 1 |
13 ± 3 |
no |
50 |
10 ± 3 |
16 ± 5 |
no |
160 |
11 ± 2 |
15 ± 4 |
no |
500 |
11 ± 1 |
21 ± 2 |
no |
1600 |
15 ± 3 |
21 ± 2 |
no |
5000 |
13 ± 1 B,P |
20 ± 4 |
no |
Positive control |
49 ± 14 |
154 ± 13 |
no |
*solvent control with THF; B = background lawn reduced; Precipitation
Table #3: Preincubation test: Number of revertants per plate (mean of 3 plates)
|
[Strain TA 98] |
[Strain TA 100] |
[Strain TA 1535] |
||||||
Conc. |
- MA |
+ MA |
Cytotoxic |
- MA |
+ MA |
Cytotoxic |
- MA |
+ MA |
Cytotoxic |
0* |
21 ± 5 |
22 ± 6 |
no |
81 ± 8 |
84 ± 9 |
no |
9 ± 2 |
6 ± 3 |
no |
50 |
24 ± 4 |
23 ± 6 |
no |
101 ± 16 |
77 ± 11 |
no |
7 ± 3 |
9 ± 2 |
no |
160 |
19 ± 4 |
17 ± 6 |
no |
88 ± 10 |
81 ± 11 |
no |
6 ± 3 |
8 ± 2 |
no |
500 |
24 ± 4 |
20 ± 3 |
no |
66 ± 2 |
82 ± 8 |
no |
4 ± 2 |
9 ± 2 |
no |
1600 |
24 ±3 |
17 ± 4 |
no |
82 ± 4 |
93 ± 15 |
no |
7 ± 3 |
9 ± 3 |
no |
5000 |
23 ± 9 |
19 ± 3 |
no |
80 ± 10 |
104 ± 20 |
no |
10 ± 6 |
11 ± 1 |
no |
Positive control |
103 ± 17 |
1059 ± 330 |
no |
539 ± 17 |
1033 ± 163 |
no |
327 ± 46 |
220 ± 49 |
no |
*solvent control with THF
Table #4: Preincubation test: Number of revertants per plate (mean of 3 plates)
|
[Strain TA 1537] |
||
Conc. |
- MA |
+ MA |
Cytoxic (yes/no) |
0* |
8 ±3 |
11 ± 4 |
no |
50 |
7 ± 2 |
9 ± 3 |
no |
160 |
6 ± 2 |
11 ± 3 |
no |
500 |
10 ± 3 |
13 ± 1 |
no |
1600 |
7 ± 1 |
14 ± 2 |
no |
5000 |
6 ± 4 |
11 ± 3 |
no |
Positive control |
55 ± 22 |
143 ± 35 |
no |
*solvent control with THF
Summary Table
relative | relative | relative | mutant | relative | relative | relative | mutant | ||||||
conc. | P | S9 | cloning | cell | cloning | colonies/ | induction | cloning | cell | cloning | colonies/ | induction | |
µg/mL | mix | efficiency I | density | efficiency II | 106cells | factor | efficiency I | density | efficiency II | 106cells | factor | ||
% | % | % | % | % | % | ||||||||
Column | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 |
Experiment I / 4 h treatment | culture I | culture II | |||||||||||
Solvent control with THF | - | 100.0 | 100.0 | 100.0 | 12.3 | 1.0 | 100.0 | 100.0 | 100.0 | 13.8 | 1.0 | ||
Positive control (EMS) | 150.0 | - | 84.1 | 132.6 | 113.8 | 70.9 | 5.8 | 80.4 | 76.7 | 63.8 | 114.8 | 8.3 | |
Test item | 4.7 | - | 97.9 | culture was not continued# | 87.8 | culture was not continued# | |||||||
Test item | 9.4 | - | 84.6 | 104.9 | 118.8 | 13.4 | 1.1 | 79.6 | 106.1 | 55.7 | 24.3 | 1.8 | |
Test item | 18.8 | - | 88.6 | 142.2 | 100.7 | 9.8 | 0.8 | 98.8 | 105.7 | 31.0 | 47.0 | 3.4 | |
Test item | 37.5 | - | 80.4 | 155.1 | 139.6 | 12.3 | 1.0 | 83.9 | 76.0 | 39.0 | 25.3 | 1.8 | |
Test item | 75.0 | P | - | 68.5 | 70.8 | 113.2 | 8.3 | 0.7 | 82.0 | 96.4 | 42.7 | 25.1 | 1.8 |
Test item | 150.0 | P | - | 80.7 | 90.3 | 126.2 | 7.4 | 0.6 | 69.8 | 80.6 | 35.6 | 84.9 | 6.1 |
Experiment I / 4 h treatment | culture I | culture II | |||||||||||
Solvent control with THF | + | 100.0 | 100.0 | 100.0 | 11.9 | 1.0 | 100.0 | 100.0 | 100.0 | 10.7 | 1.0 | ||
Positive control (DMBA) | 1.1 | + | 58.3 | 70.4 | 70.2 | 802.4 | 67.3 | 85.0 | 48.2 | 119.1 | 274.5 | 25.5 | |
Test item | 4.7 | + | 81.4 | culture was not continued# | 113.4 | culture was not continued# | |||||||
Test item | 9.4 | + | 77.8 | 120.4 | 92.1 | 28.2 | 2.4 | 96.0 | 60.9 | 109.9 | 13.3 | 1.2 | |
Test item | 18.8 | + | 80.5 | 105.1 | 100.7 | 10.3 | 0.9 | 119.3 | 87.5 | 92.3 | 17.5 | 1.6 | |
Test item | 37.5 | + | 78.5 | 130.0 | 100.3 | 11.5 | 1.0 | 119.0 | 97.9 | 105.2 | 8.5 | 0.8 | |
Test item | 75.0 | P | + | 72.4 | 100.9 | 119.0 | 9.5 | 0.8 | 128.4 | 77.6 | 109.0 | 7.4 | 0.7 |
Test item | 150.0 | P | + | 72.6 | 98.2 | 107.7 | 6.7 | 0.6 | 131.9 | 76.2 | 121.7 | 15.1 | 1.4 |
Experiment II / 24 h treatment | culture I | culture II | |||||||||||
Solvent control | - | 100.0 | 100.0 | 100.0 | 20.4 | 1.0 | 100.0 | 100.0 | 100.0 | 9.7 | 1.0 | ||
Positive control (EMS) | 150.0 | - | 108.1 | 79.6 | 86.7 | 381.1 | 18.7 | 105.1 | 126.4 | 103.5 | 422.4 | 43.7 | |
Test item | 4.7 | - | 109.1 | culture was not continued# | 106.8 | culture was not continued# | |||||||
Test item | 9.4 | - | 98.6 | 108.0 | 84.6 | 26.2 | 1.3 | 105.6 | 95.6 | 117.2 | 23.1 | 2.4 | |
Test item | 18.8 | - | 98.8 | 92.3 | 92.4 | 34.1 | 1.7 | 109.3 | 75.7 | 98.4 | 20.2 | 2.1 | |
Test item | 37.5 | P | - | 102.2 | 93.8 | 85.5 | 14.7 | 0.7 | 106.1 | 93.8 | 99.1 | 9.9 | 1.0 |
Test item | 75.0 | P | - | 97.2 | 104.0 | 84.7 | 24.0 | 1.2 | 105.8 | 144.5 | 113.3 | 13.2 | 1.4 |
Test item | 150.0 | P | - | 99.0 | 115.4 | 79.0 | 30.3 | 1.5 | 103.9 | 94.1 | 95.9 | 2.6 | 0.3 |
Experiment II / 4 h treatment | |||||||||||||
Solvent control with THF | + | 100.0 | 100.0 | 100.0 | 35.2 | 1.0 | 100.0 | 100.0 | 100.0 | 21.0 | 1.0 | ||
Positive control (DMBA) | 1.1 | + | 103.0 | 120.7 | 67.8 | 732.9 | 20.8 | 100.0 | 105.0 | 93.9 | 348.7 | 16.6 | |
Test item | 4.7 | + | 104.4 | culture was not continued# | 101.6 | culture was not continued# | |||||||
Test item | 9.4 | + | 123.9 | 109.0 | 98.6 | 14.8 | 0.4 | 108.7 | 107.6 | 95.2 | 4.5 | 0.2 | |
Test item | 18.8 | + | 133.2 | 89.6 | 93.2 | 29.6 | 0.8 | 112.2 | 109.0 | 98.7 | 10.8 | 0.5 | |
Test item | 37.5 | + | 94.0 | 127.6 | 79.8 | 26.9 | 0.8 | 100.9 | 93.4 | 101.3 | 16.4 | 0.8 | |
Test item | 75.0 | P | + | 97.8 | 95.3 | 74.5 | 23.4 | 0.7 | 105.7 | 123.3 | 107.4 | 15.7 | 0.7 |
Test item | 150.0 | P | + | 106.3 | 104.3 | 74.7 | 15.7 | 0.4 | 105.3 | 99.5 | 94.7 | 14.2 | 0.7 |
# culture was not continued since a minimum of only four analysable concentrations is required
P precipitation observed at the end of treatment
Summary Table
relative | relative | relative | mutant | relative | relative | relative | mutant | ||||||
conc. | P | S9 | cloning | cell | cloning | colonies/ | induction | cloning | cell | cloning | colonies/ | induction | |
µg/mL | mix | efficiency I | density | efficiency II | 106cells | factor | efficiency I | density | efficiency II | 106cells | factor | ||
% | % | % | % | % | % | ||||||||
Column | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 |
Experiment I / 4 h treatment | culture I | culture II | |||||||||||
Solvent control with THF | - | 100.0 | 100.0 | 100.0 | 12.3 | 1.0 | 100.0 | 100.0 | 100.0 | 13.8 | 1.0 | ||
Positive control (EMS) | 150.0 | - | 84.1 | 132.6 | 113.8 | 70.9 | 5.8 | 80.4 | 76.7 | 63.8 | 114.8 | 8.3 | |
Test item | 4.7 | - | 97.9 | culture was not continued# | 87.8 | culture was not continued# | |||||||
Test item | 9.4 | - | 84.6 | 104.9 | 118.8 | 13.4 | 1.1 | 79.6 | 106.1 | 55.7 | 24.3 | 1.8 | |
Test item | 18.8 | - | 88.6 | 142.2 | 100.7 | 9.8 | 0.8 | 98.8 | 105.7 | 31.0 | 47.0 | 3.4 | |
Test item | 37.5 | - | 80.4 | 155.1 | 139.6 | 12.3 | 1.0 | 83.9 | 76.0 | 39.0 | 25.3 | 1.8 | |
Test item | 75.0 | P | - | 68.5 | 70.8 | 113.2 | 8.3 | 0.7 | 82.0 | 96.4 | 42.7 | 25.1 | 1.8 |
Test item | 150.0 | P | - | 80.7 | 90.3 | 126.2 | 7.4 | 0.6 | 69.8 | 80.6 | 35.6 | 84.9 | 6.1 |
Experiment I / 4 h treatment | culture I | culture II | |||||||||||
Solvent control with THF | + | 100.0 | 100.0 | 100.0 | 11.9 | 1.0 | 100.0 | 100.0 | 100.0 | 10.7 | 1.0 | ||
Positive control (DMBA) | 1.1 | + | 58.3 | 70.4 | 70.2 | 802.4 | 67.3 | 85.0 | 48.2 | 119.1 | 274.5 | 25.5 | |
Test item | 4.7 | + | 81.4 | culture was not continued# | 113.4 | culture was not continued# | |||||||
Test item | 9.4 | + | 77.8 | 120.4 | 92.1 | 28.2 | 2.4 | 96.0 | 60.9 | 109.9 | 13.3 | 1.2 | |
Test item | 18.8 | + | 80.5 | 105.1 | 100.7 | 10.3 | 0.9 | 119.3 | 87.5 | 92.3 | 17.5 | 1.6 | |
Test item | 37.5 | + | 78.5 | 130.0 | 100.3 | 11.5 | 1.0 | 119.0 | 97.9 | 105.2 | 8.5 | 0.8 | |
Test item | 75.0 | P | + | 72.4 | 100.9 | 119.0 | 9.5 | 0.8 | 128.4 | 77.6 | 109.0 | 7.4 | 0.7 |
Test item | 150.0 | P | + | 72.6 | 98.2 | 107.7 | 6.7 | 0.6 | 131.9 | 76.2 | 121.7 | 15.1 | 1.4 |
Experiment II / 24 h treatment | culture I | culture II | |||||||||||
Solvent control | - | 100.0 | 100.0 | 100.0 | 20.4 | 1.0 | 100.0 | 100.0 | 100.0 | 9.7 | 1.0 | ||
Positive control (EMS) | 150.0 | - | 108.1 | 79.6 | 86.7 | 381.1 | 18.7 | 105.1 | 126.4 | 103.5 | 422.4 | 43.7 | |
Test item | 4.7 | - | 109.1 | culture was not continued# | 106.8 | culture was not continued# | |||||||
Test item | 9.4 | - | 98.6 | 108.0 | 84.6 | 26.2 | 1.3 | 105.6 | 95.6 | 117.2 | 23.1 | 2.4 | |
Test item | 18.8 | - | 98.8 | 92.3 | 92.4 | 34.1 | 1.7 | 109.3 | 75.7 | 98.4 | 20.2 | 2.1 | |
Test item | 37.5 | P | - | 102.2 | 93.8 | 85.5 | 14.7 | 0.7 | 106.1 | 93.8 | 99.1 | 9.9 | 1.0 |
Test item | 75.0 | P | - | 97.2 | 104.0 | 84.7 | 24.0 | 1.2 | 105.8 | 144.5 | 113.3 | 13.2 | 1.4 |
Test item | 150.0 | P | - | 99.0 | 115.4 | 79.0 | 30.3 | 1.5 | 103.9 | 94.1 | 95.9 | 2.6 | 0.3 |
Experiment II / 4 h treatment | |||||||||||||
Solvent control with THF | + | 100.0 | 100.0 | 100.0 | 35.2 | 1.0 | 100.0 | 100.0 | 100.0 | 21.0 | 1.0 | ||
Positive control (DMBA) | 1.1 | + | 103.0 | 120.7 | 67.8 | 732.9 | 20.8 | 100.0 | 105.0 | 93.9 | 348.7 | 16.6 | |
Test item | 4.7 | + | 104.4 | culture was not continued# | 101.6 | culture was not continued# | |||||||
Test item | 9.4 | + | 123.9 | 109.0 | 98.6 | 14.8 | 0.4 | 108.7 | 107.6 | 95.2 | 4.5 | 0.2 | |
Test item | 18.8 | + | 133.2 | 89.6 | 93.2 | 29.6 | 0.8 | 112.2 | 109.0 | 98.7 | 10.8 | 0.5 | |
Test item | 37.5 | + | 94.0 | 127.6 | 79.8 | 26.9 | 0.8 | 100.9 | 93.4 | 101.3 | 16.4 | 0.8 | |
Test item | 75.0 | P | + | 97.8 | 95.3 | 74.5 | 23.4 | 0.7 | 105.7 | 123.3 | 107.4 | 15.7 | 0.7 |
Test item | 150.0 | P | + | 106.3 | 104.3 | 74.7 | 15.7 | 0.4 | 105.3 | 99.5 | 94.7 | 14.2 | 0.7 |
# culture was not continued since a minimum of only four analysable concentrations is required
P precipitation observed at the end of treatment
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Description of key information
B6C3F1 mice (10/sex/dose) received diets containing 0, 0.62%, 1.25%, 2.5%, 5.0% or 10% castor oil, continuously for 13 weeks. Ten additional rats/sex were included at each dose level for evaluation of hematological and clinical chemistry parameters. At days 5 and 21, these animals were anesthetized with carbon dioxide, and blood was collected from the orbital sinus. These animals were killed following the blood collection on day 21.Smears were prepared from peripheral blood samples obtained by cardiac puncture of dosed and control animals at the termination of the 13 week study. Slides were stained with Hoechst 33258/pyronin Y. At least 2000 PCE and 10000 NCE from each animal were scored for micronuclei. Castor oil was found to be negative for cytogenicity as evidenced by a lack of micronuclei in peripheral blood erythrocytes of B6C3F1 mice exposed in dosed feeding for 13 weeks.
Link to relevant study records
- Endpoint:
- in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
- 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
- Qualifier:
- no guideline followed
- Principles of method if other than guideline:
- - Principle of test:
Mice (10/sex/dose) received diets containing 0, 0.62%, 1.25%, 2.5%, 5.0% or 10% castor oil, continuously for 13 weeks. Ten additional rats/sex were included at each dose level for evaluation of hematological and clinical chemistry parameters. At days 5 and 21, these animals were anesthetized with carbon dioxide, and blood was collected from the orbital sinus. These animals were killed following the blood collection on day 21.Smears were prepared from peripheral blood samples obtained by cardiac puncture of dosed and control animals at the termination of the 13 week study. Slides were stained with Hoechst 33258/pyronin Y. At least 2000 PCE and 10000 NCE from each animal were scored for micronuclei.
- Short description of test conditions: Mice were housed individually. Polycarbonate cages lined with heat-treated hardwood chips and covered with polyester filter sheets were used; the cages were stored on stainless steel racks equipped with an automatic watering system. Temperature in the animal room was maintained within 68-76°F; relative humidity ranged from 42% to 72%. Incoming air was filtered to remove particulates, and a flow rate was maintained to ensure complete exchange at least 10 times per hour. A controlled light cycle of 12 hours of daylight and 12 hours of darkness was maintained. Control feed or diet formulations of castor oil were available ad libitum; feeders were changed twice per week throughout the study.
- Parameters analysed / observed: Smears were prepared from peripheral blood samples obtained by cardiac puncture of dosed and control animals at the termination of the 13 week study. Slides were stained with Hoechst 33258/pyronin Y. At least 2000 PCE and 10000 NCE from each animal were scored for micronuclei. - GLP compliance:
- yes
- Type of assay:
- mammalian erythrocyte micronucleus test
- Specific details on test material used for the study:
- USP AA grade castor oil was obtained in one lot (#L-5G30-01) from Cas Chemical, Inc. (Bayonne, NJ). Purity and identity analyses were conducted by Midwest Research Institute (MRI) (Kansas City, MO). MRI reports on the analyses performed in support of the castor oil studies are on file at the National Institute of Environmental Health Sciences (Research Triangle Park, NC).
Cumulative data indicated a purity consistent with the USP specifications and the reported composition for castor oil.
The stability of the study material during the toxicology studies was monitored by determination of peroxide content and by high performance liquid chromatography. No deterioration of the castor oil study material was observed over the course of the studies. - Species:
- mouse
- Strain:
- B6C3F1
- Details on species / strain selection:
- Obtained from Simonsen Laboratories (Gilroy, CA, USA).
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- Mice were housed individually. Polycarbonate cages lined with heat-treated hardwood chips and covered with polyester filter sheets were used; the cages were stored on stainless steel racks equipped with an automatic watering system. Temperature in the animal room was maintained within 68-76°F; relative humidity ranged from 42% to 72%. Incoming air was filtered to remove particulates, and a flow rate was maintained to ensure complete exchange at least 10 times per hour. A controlled light cycle of 12 hours of daylight and 12 hours of darkness was maintained. Control feed or diet formulations of castor oil were available ad libitum; feeders were changed twice per week throughout the study.
- Route of administration:
- oral: feed
- Details on exposure:
- Ad libitum.
- Duration of treatment / exposure:
- 13 continuous weeks.
- Frequency of treatment:
- Daily
- Dose / conc.:
- 0 other: %
- Dose / conc.:
- 0.62 other: %
- Dose / conc.:
- 1.25 other: %
- Dose / conc.:
- 2.5 other: %
- Dose / conc.:
- 5 other: %
- Dose / conc.:
- 10 other: %
- No. of animals per sex per dose:
- 10/sex/concentration.
- Control animals:
- yes, plain diet
- Positive control(s):
- Male mice treated for 4 weeks with urethane in the drinking water (0.2%). These animals were not part of the 13-week study, but were added as a measure of quality control for the assay.
- Tissues and cell types examined:
- Complete histopathology examinations were conducted on all mice from the control and 10% dose groups.
- Details of tissue and slide preparation:
- Smears were prepared from peripheral blood samples obtained by cardiac puncture of dosed and control animals at the termination of the 13 week study. Slides were stained with Hoechst 33258/pyronin Y. At least 2000 PCE and 10000 NCE from each animal were scored for micronuclei.
- Statistics:
- Shirley's test was used to assess any significance that were different from control groups by , p <0.05.
- Key result
- Sex:
- male/female
- Genotoxicity:
- negative
- Toxicity:
- no effects
- Vehicle controls validity:
- not applicable
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Conclusions:
- Castor oil was found to be negative for cytogenicity as evidenced by a lack of micronuclei in peripheral blood erythrocytes of B6C3F1 mice exposed in dosed feeding for 13 weeks.
- Executive summary:
B6C3F1 mice (10/sex/dose) received diets containing 0, 0.62%, 1.25%, 2.5%, 5.0% or 10% castor oil, continuously for 13 weeks. Ten additional rats/sex were included at each dose level for evaluation of hematological and clinical chemistry parameters. At days 5 and 21, these animals were anesthetized with carbon dioxide, and blood was collected from the orbital sinus. These animals were killed following the blood collection on day 21.Smears were prepared from peripheral blood samples obtained by cardiac puncture of dosed and control animals at the termination of the 13 week study. Slides were stained with Hoechst 33258/pyronin Y. At least 2000 PCE and 10000 NCE from each animal were scored for micronuclei. Castor oil was found to be negative for cytogenicity as evidenced by a lack of micronuclei in peripheral blood erythrocytes of B6C3F1 mice exposed in dosed feeding for 13 weeks.
Reference
Frequency of Micronuclei in Peripheral Blood Erythrocytes of B6C3F1 Mice Exposed to Castor Oil in Dosed Feed for 13 Weeks
Percent in Feed | % Normachromatic erythrocytes with micronuclei | % Polychromatic erythrocytes with micronuclei | Number of mice |
Male mice: | |||
0 | 0.11 ± 0.02 | 1.20 ± 0.08 | 10 |
0.6 | 0.13 ± 0.02 | 1.18 ± 0.10 | 10 |
1.3 | 0.11 ± 0.01 | 1.16 ± 0.09 | 10 |
2.5 | 0.13 ± 0.01 | 1.24 ± 0.10 | 10 |
5.0 | 0.09 ± 0.02 | 1.40 ± 0.11 | 9 |
10 | 0.09 ± 0.01 | 1.21 ± 0.08 |
10 |
Female mice | |||
0 | 0.10 ± 0.01 | 1.18 ± 0.07 | 10 |
0.6 | 0.09 ± 0.01 | 1.21 ± 0.10 | 10 |
1.3 | 0.07 ± 0.01 | 1.11 ± 0.08 | 9 |
2.5 | 0.09 ± 0.02 | 1.11 ± 0.08 | 10 |
5.0 | 0.09 ± 0.01 | 1.49 ± 0.18 | 10 |
10.0 | 0.06 ± 0.01 | 1.00 ± 0.10 | 10 |
Urethane | |||
0.2% | 1.68 ± 0.25 | 1.710 ± 0.25 | 3 |
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
Justification for classification or non-classification
Based on the findings of a number of different reliable in vitro and in vivo mutagenicity studies conducted on the substance and structural analogues, classification of the substance is not justified.
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.