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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 297-627-7 | CAS number: 93685-79-1
- 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
There is no in vitro genetic toxicity data available for isoeicosane. However, data is available for structural analogues isohexadecane, hydrocarbons, C12-C16, n-alkanes, isoalkanes, cyclics, <2% aromatics, and hydrodesulfurized kerosene. This data is read across to based on analogue read across and a discussion and report on the read across strategy is provided as an attachment in IUCLID Section 13.
All read across genetic toxicity tests listed below had negative results for isoeicosane.
Genetic Toxicity in vitro – Bacterial reverse mutation assay (OECD 471)
Genetic Toxicity in vitro – mammalian chromosome aberration test (OECD TG 473)
Genetic Toxicity in vitro - Mammalian Cell Gene Mutation Test (OECD TG 476)
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Study period:
- no data
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: GLP study, the protocol and the results were described with details. Substance analytical certificate not available
- Justification for type of information:
- A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
- Reason / purpose for cross-reference:
- read-across: supporting information
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- yes
- Remarks:
- only 4 bacterial strains were used instead of 5
- Principles of method if other than guideline:
- Guideline study
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- Reversion Histidine auxotrophy
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Details on mammalian cell type (if applicable):
- Not applicable
- Additional strain / cell type characteristics:
- not applicable
- Remarks:
- The strain cultures were stored as stock cultures in ampoules with nutrient broth + 5% DMSO in liquid nitrogen.
- Metabolic activation:
- with and without
- Metabolic activation system:
- The S9 liver microsomal fraction was obtained from the liver of 8-12 weeks old male wistar rats; strain WU (Savo-Ivanovas, med. Versuchstier Zuchten GmbH) which received a single i.p. injection of 500 mg/kg b.w. Aroclor 1254 in olive oil 5 days previously
- Test concentrations with justification for top dose:
- 10.0; 100.0; 333.3; 1000 and 5000 µg/plate (With and without S9 mix)
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: ethanol
- Justification for choice of solvent/vehicle: the solvent was chosen to its solubility properties and its relative non-toxicity for the bacteria. - Untreated negative controls:
- yes
- Remarks:
- concurrent untreated control
- Negative solvent / vehicle controls:
- yes
- Remarks:
- ethanol
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: See below Table 7.6.1/2
- Remarks:
- 6 plate for negative control (untreated strains), 6 plate for solvent, 3 plates for positive controls.
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation)
After range-finding test, two independent experiments were conducted in the main test by agar plate incorporation with and without S9 mix. The different controls (negative, solvent and positive controls) were tested in the same conditions.
DURATION
- Preincubation period: the bacterial culture was incubated in a shaking water bath for 6 hours at 37°C.
- Exposure duration: 3 days at 37°C in the dark
SELECTION AGENT (mutation assays):histidine
NUMBER OF REPLICATION: three scoring (3 measurements/plate), the mean number and standard deviation of revertants are calculated for all groups. the means for all treatment groups are compared with those obtained for the solvent control groups.
DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency; relative total growth
Other: the colonies were counted using the BIOTRAN III counter. If precipitation of the test article precluded automatic counting the revertant colonies were counted by hand. - Evaluation criteria:
- - A positive response was indicated by a reproducible, dose-related increase, whether it be two-fold over background or not.
- Mutation Factors (MF) (induced/spontaneous revertants) were calculated for all strains at the dose level tested. - Statistics:
- A compound is deemed to provide evidence of mutagenic potential if:
- a statistically significant dose-related increase in the number of revertant colonies is obtained in two separate experiments, and
- the increase in the number of revertant colonies is at least twice the concurrent solvent control value. - 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
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- not applicable
- Genotoxicity:
- not determined
- Cytotoxicity / choice of top concentrations:
- not determined
- Vehicle controls validity:
- not examined
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- not examined
- Additional information on results:
- See tables 7.6.1/3, 7.6.1/4, 7.6.1/5 and 7.6.1/6
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: no data
- Effects of osmolality: no data
- Evaporation from medium: no data
- Water solubility: no data
- Precipitation: no data
- Other confounding effects: no data
RANGE-FINDING/SCREENING STUDIES:
A preliminary toxicity assay was conducted with strains TA98 and TA100 at concentrations between 1 and 5000 µg/plate. The plates with the test article showed normal background growth up to 5000.0 µg/plate in strain TA98 and TA100.
COMPARISON WITH HISTORICAL CONTROL DATA: no
ADDITIONAL INFORMATION ON CYTOTOXICITY: no - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results:
negative TA1535; TA1537; TA98 and TA 100
Under the conditions of the assay, Isohexadecan did not demonstrate in vitro mutagenic activity in the Salmonella test system with and without S9 mix activation system. - Executive summary:
In a reverse gene mutation assay in bacteria (Poth, 1990) and in compliance with Good Laboratory Practice, strains TA98, TA100, TA1535 and TA1537 of S. typhimurium were exposed to Isohexadecan at concentrations of 10.0, 100.0, 333.3, 1000.0 and 5000.0 µg/plate in the presence and absence of mammalian metabolic activation. No cytotoxicity was observed with all the dose tested. Up to the highest investigated dose, no significant and reproducible dose-dependent increase in revertant colony numbers was obtained in any of the Salmonella typhimurium strains used (+/- S9). The positive controls induced the appropriate responses in the corresponding strains. Under the test conditions, Isohexadecan did not induce in vitro mutagenic activity in the bacterial test system in the presence and the absence of S9 activation system.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Study period:
- 10 August to 21 September 1990
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Few details on test material (no certificate of analysis)
- Justification for type of information:
- A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
- Reason / purpose for cross-reference:
- read-across: supporting information
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- yes
- Remarks:
- No certificate of analysis
- Principles of method if other than guideline:
- Guideline principles
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- in vitro mammalian chromosome aberration test
- Target gene:
- Not applicable
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Details on mammalian cell type (if applicable):
- - Type and identity of media: McCoy's 5A culture medium supplemented with 10% fetal calf serum, 1% L-glutamine, and 1% penicillin and streptomycin, at about 37°C, in an atmosphere of about 5% C02 in air.
- Properly maintained: yes - Additional strain / cell type characteristics:
- not applicable
- Metabolic activation:
- with and without
- Metabolic activation system:
- Rat liver S9 from male Sprague-Dawley rats treated with Aroclor 1254
- Test concentrations with justification for top dose:
- Range finding assay: half-log series of concentrations of 0.0835 to 2500 µg/mL
Main experiment:
- without metabolic activation: 3.13, 6.26, 9.35 and 12.5 µg/mL with 10-h harvest and 12.5, 25, 37.5, 50 and 75 µg/mL with 20-h harvest
- with metabolic activation: 37.5, 93.8, 188, 281, 375, 563 and 750 µg/mL for 10 and 20-h harvest - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: ethanol
- Justification for choice of solvent/vehicle: test material was insoluble in water and dimethylsulfoxide. A clear and homogeneous stock solution of 201 mg/mL with ethanol could be maintained. - Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: See below
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Preincubation period: 24 h
- Exposure duration: without metabolic activation: 7.25 and 17 h for 10 and 20 h assay, respectively; with metabolic activation: 2 h
- Expression time (cells in growth medium): with metabolic activation: 7.75 and 17.75 h for 20 and 10 h assay, respectively;
- Time in 0.1 µg/mL Colcemid: without metabolic activation: 1 and 0.5 h for 20 and 10 h assay, respectively; with metabolic activation: 2.5 h
- Fixation time (start of exposure up to fixation or harvest of cells): 10 h and 20 h without and without metabolic activation
STAIN (for cytogenetic assays): 5% Giemsa solution and BrdUrd (5-bromodeoxyuridine) at 10 µM
NUMBER OF REPLICATIONS: 2
NUMBER OF CELLS EVALUATED: 100 cells for test substance; at least 25 cells for positive controls
CYTOTOXICITY: visual observations based on confluence of monolayer and floating dead cells - Evaluation criteria:
- Cells were selected for good morphology and only cells with the number of centromeres equal to the modal number 21 ± 2 were analyzed.
The following factors were taken into account in the evaluation of the chromosomal aberrations data: the overall chromosomal aberration frequencies, the percentage of cells with any aberrations, the percentage of cells with more than one aberration, any evidence for increasing amounts of damage with increasing dose.
Chromatid and isochromatid gaps were not considered as they may be due to toxicity. - Statistics:
- Fisher's exact test with an adjustment of multiple comparisons
- Key result
- Species / strain:
- Chinese hamster Ovary (CHO)
- 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:
- Range-finding without metabolic activation:
A very unhealthy cell monolayer, -70% reduction in the cell monolayer confluence, floating dead cells, and severe reduction in the number of visible mitotic cells were observed in the culture dosed with 25.0 µg/mL. Slight reductions in the number of visible mitotic cells and -15% reduction in the cell monolayer confluence were observed in the cultures dosed with 2.50 and 8.35 µg/mL.
Range-finding with metabolic activation:
An unhealthy cell monolayer, -85% reduction in the cell monolayer confluence, floating dead cells and debris, and severe reduction in the number of visible mitotic cells were observed in the culture dosed with 835 µg/mL. Reductions of -15% in the cell monolayer confluence were observed in the cultures dosed with 25.0 and 83.5 µg/mL.
Chromosomal aberrations assay without metabolic activation (Table 1):
In the 10 h assay, no toxicity was observed in any of the test cultures. These cultures were not analyzed for chromosomal aberrations as four dose levels were available for analysis from the 20 h assay. In the 20 h assay, an unhealthy cell monolayer, -70% and -45 % reduction in the cell monolayer confluence, floating dead cells and debris, and a severe reduction in visible mitotic cells were observed at 75.0 and 50.0 µg/mL, respectively. Toxicity was evident on the slides prepared from these cultures by the very sparse numbers of metaphases available for analysis.
Chromosomal aberration assay with metabolic activation (Tables 2 and 3):
In the 10 h assay, slight reductions in the numbers of visible mitotic cells were observed in the cultures dosed at 563 and 751 µg/mL.
In the 20 h assay, severe toxicity was exhibited on the slides prepared from the cultures dosed with 562 and 750 µg/mL by the presence of many dead cells and the sparse numbers of metaphases available for analysis. Reductions of -15% in the cell monolayer confluence were observed in the cultures dosed with 99.7, 187, 281, 375, 562, and 750 µg/mL. - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results:
negative
MRD-90-843 was found not to increase chromosome aberrations in CHO cells with and without metabolic activation. - Executive summary:
In an in vitro chromosome aberration test, Chinese Hamster Ovary cells were exposed to MRD-90-843 at concentrations of 3.13, 6.26, 9.35 and 12.5 µg/mL for 10-h harvest and 12.5, 25, 37.5, 50 and 75 µg/mL for 20-h harvest, for 7 and 17 h, without metabolic activation and 37.5, 93.8, 188, 281, 375, 563 and 750 µg/mL for 10 and 20-h harvest, for 2 h, with metabolic activation.
Positive controls (mitomycin C without metabolic activation and cyclophosphamide with metabolic activation) induced the appropriate response. As there was no evidence of chromosome aberration induced over background, MRD-90-843 is not classified according to the criteria of Annex VI to Directive 67/548/EEC and the CLP Regulation (1272/2008).
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Study period:
- 1984
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Equivalent or similar to OECD Guideline 476.
- Justification for type of information:
- A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
- Reason / purpose for cross-reference:
- read-across: supporting information
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- GLP compliance:
- not specified
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- TK locus
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- na
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 homogenate
- Test concentrations with justification for top dose:
- Without activation: 6.25 nl/ml to 37.5 nl/ml
With activation: 3.91 nl/ml to 62.5 nl/ml - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: [ethanol]
- Justification for choice of solvent/vehicle:The test material was miscible with ethanol. - Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- ethylmethanesulphonate
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 4 hours
- Expression time (cells in growth medium): 24 hours
SELECTION AGENT (mutation assays): BrdU
NUMBER OF REPLICATIONS: Variable with or without activation
NUMBER OF CELLS EVALUATED: 3x10^6
DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency - Key result
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- None of the assayed treatments induced a mutant frequency that exceeded the minimum criterion of 40.8 x 10^-6
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- The test material induced a good range of toxicities for evaluation of the test material (percent relative growths, 65.3% to 2.8%). The toxicities did show some variability between replicate samples. In the presence of metabolic activation, no indication of mutagenic activity was observed. The average cloning efficencies for the solvent and untreated negative controls varied from 119.1% without activation to 82.7% with activation which demonstrated very good cloning conditions for the assays. The negative control mutant frequencies were all in the normal range and the positive compounds yielded normal mutant frequencies that were greatly in excess of the background.
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results:
negative with metabolic activation
negative without metabolic activation
It is concluded in this study that the test material is not a mutagenic agent with or without activation. Classification is not warranted under the new Regulation (EC) 1272/2008 on classification, labeling, and packaging of substances and mixtures (CLP) or under the Directive 67/518/EEC for dangerous substances and Directive 1999/45/EC for preparations. - Executive summary:
The test material was examined for mutagenic activity in the mouse lymphoma forward mutation assay in the absence and presence of a liver S9 fraction for metabolic activation. The test material did not induce significant increases in the mutant frequency at the TK locus in L5178Y mouse lymphoma cells. Treatments up to 37.5 nl/ml without activation and 62.5 nl/ml with activation were assayed and high toxicities were induced without inducing significant increases in the mutant frequency. It is concluded in this study that the test material is not a mutagenic agent with or without activation. Classification is not warranted under the new Regulation (EC) 1272/2008 on classification, labeling, and packaging of substances and mixtures (CLP) or under the Directive 67/518/EEC for dangerous substances and Directive 1999/45/EC for preparations.
Referenceopen allclose all
Table 7.6.1/3: Number of revertants per plate (mean of triplicates) in the absence of metabolic activation (First test)
Test substance concentration |
TA 1535 |
TA 1537 |
TA 98 |
TA 100 |
||||||||
Mean |
Standard deviation |
Factor + |
Mean |
Standard deviation |
Factor + |
Mean |
Standard deviation |
Factor + |
Standard deviation |
Factor + |
||
Negative* control |
12 |
1.2 |
- |
8 |
0.6 |
- |
16 |
1.0 |
- |
77 |
10.0 |
- |
Solvent control** |
10 |
0.6 |
1.0 |
7 |
0.6 |
1.0 |
19 |
5.1 |
1.0 |
83 |
7.4 |
1.0 |
10 |
10 |
2.6 |
1.0 |
8 |
1.7 |
1.0 |
14 |
6.4 |
0.7 |
78 |
8.5 |
0.9 |
100 |
11 |
3.8 |
1.1 |
8 |
1.7 |
1.1 |
16 |
2.1 |
0.8 |
73 |
12.1 |
0.9 |
333.3 |
10 |
4.4 |
1.0 |
7 |
1.5 |
1.0 |
19 |
4.2 |
1.0 |
78 |
12.4 |
0.9 |
1000.0 |
11 |
3.6 |
1.1 |
7 |
1.0 |
17 |
1.2 |
0.9 |
79 |
4.7 |
0.9 |
|
5000.0 |
13 |
4.6 |
1.3 |
5 |
0 |
0.7 |
22 |
2.9 |
1.1 |
73 |
7.4 |
0.9 |
Positive control*** |
839 |
20.2 |
86.8 |
228 |
9.8 |
31.1 |
1694 |
244.4 |
87.6 |
1065 |
48.5 |
12.8 |
Negative control*: concurrent untreated control
Solvent control**: ethanol
Positive control***: see table 7.6.1/2
Table 7.6.1/4: Number of revertants per plate (mean of triplicates) in the presence of metabolic activation (First test)
Test substance concentration |
TA 1535 |
TA 1537 |
TA 98 |
TA 100 |
||||||||
Mean |
Standard deviation |
Factor + |
Mean |
Standard deviation |
Factor + |
Mean |
Standard deviation |
Factor + |
Mean |
Standard deviation |
Factor + |
|
Negative control* |
10 |
3.1 |
- |
9 |
1.5 |
- |
32 |
2.0 |
- |
91 |
10.4 |
- |
Solvent control** |
11 |
2.6 |
1.0 |
7 |
3.0 |
1.0 |
38 |
6.1 |
1.0 |
85 |
8.5 |
1.0 |
10 |
12 |
4.7 |
1.1 |
11 |
4.4 |
1.6 |
26 |
5.3 |
0.7 |
77 |
5.5 |
0.9 |
100 |
12 |
2.0 |
1.1 |
8 |
3.5 |
1.2 |
20 |
4.0 |
0.5 |
72 |
5.0 |
0.8 |
333.3 |
12 |
3.8 |
1.1 |
8 |
2.0 |
1.1 |
26 |
0.6 |
0.7 |
73 |
13.6 |
0.9 |
1000.0 |
10 |
3.0 |
0.9 |
12 |
4.0 |
1.7 |
28 |
5.0 |
0.7 |
81 |
7.6 |
1.0 |
5000.0 |
12 |
3.2 |
1.1 |
11 |
1.7 |
1.6 |
36 |
6.7 |
0.9 |
80 |
5.5 |
0.9 |
Positive control*** |
284 |
11.1 |
25.8 |
252 |
28.0 |
36.0 |
1916 |
60.5 |
50.4 |
1681 |
150.2 |
19.7 |
Negative control*:concurrent untreated control
Solvent control** ethanol
Positive control***: see Table 7.6.1/2
Table 7.6.1/5: Number of revertants per plate (mean of triplicates) in the absence of metabolic activation (second test)
Test substance concentration |
TA 1535 |
TA 1537 |
TA 98 |
TA 100 |
||||||||
Mean |
Standard deviation |
Factor + |
Mean |
Standard deviation |
Factor + |
Mean |
Standard deviation |
Factor + |
Mean |
Standard deviation |
Factor + |
|
Negative control* |
6 |
1.2 |
- |
5 |
1.0 |
- |
15 |
0.6 |
- |
80 |
11.0 |
- |
Solvent control** |
9 |
2.6 |
1.0 |
5 |
0.6 |
1.0 |
16 |
2.1 |
1.0 |
86 |
9.6 |
1.0 |
10 |
9 |
2.5 |
1.1 |
4 |
1.5 |
0.9 |
11 |
1.2 |
0.7 |
73 |
8.9 |
0.8 |
100 |
7 |
2.9 |
1.1 |
4 |
1.2 |
0.9 |
12 |
4.5 |
0.5 |
74 |
4.0 |
0.9 |
333.3 |
8 |
2.6 |
1.1 |
5 |
2.1 |
1.0 |
14 |
2.6 |
0.7 |
74 |
11.1 |
0.9 |
1000.0 |
7 |
1.0 |
0.9 |
4 |
0.6 |
0.9 |
14 |
2.6 |
0.7 |
82 |
4.7 |
1.0 |
5000.0 |
7 |
1.2 |
1.1 |
4 |
1.5 |
0.8 |
18 |
2.6 |
0.9 |
72 |
6.0 |
0.8 |
Positive control*** |
675 |
33.6 |
75.0 |
162 |
27.2 |
34.6 |
1068 |
370.0 |
65.4 |
967 |
150.2 |
11.2 |
Negative control*: concurrent untreated control
Solvent control**: ethanol
Positive control***: see table 7.6.1/2
Table 7.6.1/6: Number of revertants per plate (mean of triplicates) in the presence of metabolic activation (second test)
Test substance concentration |
TA 1535 |
TA 1537 |
TA 98 |
TA 100 |
||||||||
Mean |
Standard deviation |
Factor + |
Mean |
Standard deviation |
Factor + |
Mean |
Standard deviation |
Factor + |
Mean |
Standard deviation |
Factor + |
|
Negative control* |
7 |
3.5 |
- |
5 |
1.0 |
- |
27 |
1.2 |
- |
82 |
9.6 |
- |
Solvent control** |
6 |
1.5 |
1.0 |
10 |
1.2 |
1.0 |
34 |
6.9 |
1.0 |
86 |
4.5 |
1.0 |
10 |
9 |
2.6 |
1.4 |
7 |
1.0 |
0.7 |
29 |
1.7 |
0.9 |
70 |
3.2 |
0.8 |
100 |
6 |
3.1 |
0.9 |
6 |
1.5 |
0.7 |
35 |
3.0 |
1.0 |
79 |
8.5 |
0.9 |
333.3 |
6 |
1.2 |
1.0 |
4 |
1.2 |
0.4 |
32 |
7.2 |
0.9 |
83 |
10.1 |
1.0 |
1000.0 |
7 |
3.5 |
1.2 |
6 |
2.1 |
0.6 |
31 |
6.0 |
0.9 |
81 |
9.2 |
0.9 |
5000.0 |
6 |
1.0 |
0.9 |
6 |
1.5 |
0.7 |
31 |
6.8 |
0.9 |
76 |
6.7 |
0.9 |
Positive control*** |
199 |
9.8 |
31.4 |
211 |
42.7 |
21.8 |
1250 |
102.1 |
36.8 |
1515 |
116.3 |
17.7 |
Negative control*: concurrent untreated control
Solvent control**: ethanol
Positive control***: see table 7.6.1/2
Table 1: Chromosome aberrations in CHO cells fixed 20 h after exposure to MRD-90-843 without metabolic activation (results from pooled duplicate cultures)
|
Number and type of aberration |
|
||||
|
|
Not computed |
Simple |
Complex |
% cells with aberrations |
|
|
Concentration (µg/mL) |
Chromatid gap |
Chromosome gap |
|
|
|
Negative (vehicle) |
- |
7 |
1 |
|
|
0.0 |
Positive (Mitomycin C) |
0.04 |
7 |
|
4 |
7 |
28.0* |
Test article |
25.0 |
15 |
2 |
|
|
0.0 |
37.5 |
7 |
3 |
1 |
1 |
0.5 |
|
50.0 |
8 |
1 |
|
1 |
0.5 |
|
75.0 |
19 |
2 |
4 |
|
0.5 |
* Significantly greater than the pooled negative and vehicle controls, p<0.01
Table 2: Chromosome aberrations in CHO cells fixed 10 h after exposure to MRD-90-843 with metabolic activation (results from pooled duplicate cultures)
|
Concentration (µg/mL) |
Number and type of aberration |
|
|||
|
|
Not computed |
Simple |
Complex |
% cells with aberrations |
|
|
|
Chromatid gap |
Chromosome gap |
|
|
|
Negative (vehicle) |
- |
2 |
|
|
0.0 |
|
Positive (Cyclophosphamide) |
25.0 |
1 |
|
8 |
13 |
44.0* |
Test article |
282 |
7 |
1 |
|
|
0.0 |
375 |
3 |
1 |
0.5 |
|||
563 |
4 |
|
1 |
0.5 |
||
751 |
3 |
3 |
|
1.0 |
* Significantly greater than the pooled negative and vehicle controls, p<0.01
Table 3: Chromosome aberrations in CHO cells fixed 20 h after exposure to MRD-90-843 with metabolic activation (results from pooled duplicate cultures)
|
Concentration (µg/mL) |
Number and type of aberration |
|
|||
|
|
Not computed |
Simple |
Complex |
% cells with aberrations |
|
|
|
Chromatid gap |
Chromosome gap |
|
|
|
Negative (vehicle) |
- |
7 |
1 |
1 |
0.0 |
|
Positive (Cyclophosphamide) |
12.5 |
1 |
|
17 |
31 |
80.0* |
Test article |
281 |
15 |
2 |
|
1 |
1.0 |
375 |
16 |
6 |
1 |
1 |
1.0 |
|
562 |
3 |
1 |
1 |
1 |
1.0 |
|
750 |
10 |
1 |
1 |
1.0 |
* Significantly greater than the pooled negative and vehicle controls, p<0.01
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Description of key information
There is no in vivo genetic toxicity data available for isoeicosane. However, data is available for structural analogue hydrodesulfurized kerosene and isread across to based on a worst case basis. Adiscussion and report on the read across strategy is provided as an attachment in IUCLID Section 13.
Hydrodesulfurized kerosene was non-mutagenic when tested in an in vivo mammalian bone marrow chromosome aberration test.
Link to relevant study records
- Endpoint:
- in vivo mammalian somatic cell study: cytogenicity / bone marrow chromosome aberration
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Study period:
- 1984
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: According to or similar to guideline study OECD 475: GLP.
- Justification for type of information:
- A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
- Reason / purpose for cross-reference:
- read-across: supporting information
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 475 (Mammalian Bone Marrow Chromosome Aberration Test)
- GLP compliance:
- yes
- Type of assay:
- chromosome aberration assay
- Species:
- rat
- Strain:
- Sprague-Dawley
- Sex:
- male/female
- Route of administration:
- intraperitoneal
- Details on exposure:
- A pilot study was carried out in 4 male and 4 female young adult Sprague Dawley rats. These animals were given a single intraperitoneal (i.p.) dose (3 g/kg) of API 81-07. During the following 48 hours observation, no animals died. The doses selected for the cytogenetics study were therefore 0.3, 1 and 3 g/kg. Three groups of 15 male and 15 female rats were given a single i.p. dose of either 0.3, 1 or 3 g API 81-07/kg. At six, 24 and 48 hours after dosing 5 males and 5 females were killed at each dose level. An additional 15 males and 15 females were untreated and served as negative controls. These animals were otherwise treated the same as the test animals. A positive control group of 5 males and 5 females was administered 0.8 mg/kg Triethylenemelamine (TEM) as a single i.p. dose. These positive control animals were killed 24 hours after administration of the positive control substance. Three hours prior to being killed with CO2, animals were injected i.p. with 4 mg/kg of colchicine. After the animal was killed, the adhering soft tissue and epiphyses of both tibiae were removed and the marrow was flushed from the bone and transferred to Hank's balanced salt solution. The marrow button was collected by centrifugation and was then re suspended in 0.075M KCl. The centrifugation was repeated and the pellet resuspended in fixative (methanol:acetic acid, 3:1). The fixative was changed once and left overnight. Cells in fixative were dropped onto glass slides which were then air dried and stained with Giemsa. Slides were coded and scored for chromosomal aberrations. 50 spreads were read for each animal where feasible. A mitotic index based on at least 500 counted cells was also recorded. The index was calculated by scoring the number of cells in mitosis per 500 cells on each read slide.
- Duration of treatment / exposure:
- Three groups of 15 male and 15 female rats were given a single i.p. dose of either 0.3, 1 or 3 g API 81-07/kg. At six, 24 and 48 hours after dosing 5 males and 5 females were killed at each dose level. An additional 15 males and 15 females were untreated and served as negative controls.
- Frequency of treatment:
- Single i.p. dose of either 0.3, 1 or 3 g API 81-07/kg
- Remarks:
- Doses / Concentrations:
0, 0.3, 1 or 3 g/kg.
Basis:
analytical conc.
i.p. - No. of animals per sex per dose:
- 15 male and 15 female rats
- Control animals:
- yes, concurrent no treatment
- Positive control(s):
- These animals were otherwise treated the same as the test animals. A positive control group of 5 males and 5 females was administered 0.8 mg/kg Triethylenemelamine (TEM) as a single i.p. dose. These positive control animals were killed 24 hours after administration of the positive control substance.
- Details of tissue and slide preparation:
- Three hours prior to being killed with CO 2 , animals were injected i.p. with 4 mg/kg of colchicine. After the animal was killed, the adhering soft tissue and epiphyses of both tibiae were removed and the marrow was flushed from the bone and transferred to Hank's balanced salt solution. The marrow button was collected by centrifugation and was then resuspended in 0.075M KCl. The centrifugation was repeated and the pellet re suspended in fixative (methanol:acetic acid, 3:1). The fixative was changed once and left overnight. Cells in fixative were dropped onto glass slides which were then air dried and stained with Giemsa. Slides were coded and scored for chromosomal aberrations. 50 spreads were read for each animal where feasible. A mitotic index based on at least 500 counted cells was also recorded. The index was calculated by scoring the number of cells in mitosis per 500 cells on each read slide.
- Evaluation criteria:
- Data interpretation and evaluation Gaps were not counted as significant aberrations. Open breaks were considered as indicators of genetic damage as were configurations resulting from the repair of breaks. The latter included translocations, multiradials, rings, multicentrics, etc. Reunion figures such as these were weighed slightly higher than breaks since they usually resulted from more than one break. Cells with more than one aberration were considered to indicate more genetic damage than those with evidence of single events. Consistent variations from the euploid number were also considered in the evaluation of mutagenic potential.
The type of aberration, its frequency and its correlation to dose in a given time was considered in evaluating the test material as being positive or negative. - Statistics:
- Statistical evaluation Performed by Student's t-tests on four parameters:
1. Number of structural aberrations per animal
2. Number of numerical aberrations per animal
3. % cells with one or more structural aberrations per animal
4. % cells with 2 or more structural aberrations per animal. - Key result
- Sex:
- male/female
- Genotoxicity:
- negative
- Toxicity:
- not examined
- Vehicle controls validity:
- not applicable
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- The data are given in the report for males, females and as male and female pooled data. When the results for males were compared with those for controls and the females were compared to controls, no statistically significant differences were found. The data summarized below, are the pooled data for males and females. The structural aberration frequency did not differ significantly from the negative control at any tested dose. The percentage of cells showing one or more structural aberrations or 2 or more structural aberrations were also similar to the negative controls. A concurrent positive control group induced significant increases in aberrations.
- Conclusions:
- Interpretation of results: negative
The test material did not cause chromosome aberration in the test model. - Executive summary:
The test material did not cause chromosome aberration in the test model.
Reference
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
In Vitro
In vitro gene mutation study in bacteria
In a reverse gene mutation assay in bacteria (EC Erdolchemie, 1990) strains TA98, TA100, TA1535 and TA1537 of S. typhimurium were exposed to isohexadecane at concentrations of 10.0, 100.0, 333.3, 1000.0 and 5000.0 µg/plate in the presence and absence of mammalian metabolic activation. No cytotoxicity was observed with all the dose tested. Up to the highest investigated dose, no significant and reproducible dose-dependent increase in revertant colony numbers was obtained in any of the Salmonella typhimurium strains used (+/- S9). The positive controls induced the appropriate responses in the corresponding strains. Under the test conditions, isohexadecane did not induce in vitro mutagenic activity in the bacterial test system in the presence and the absence of S9 activation system.
In Vitro Chromosome Aberration in Mammalian Cells
In an in vitro chromosome aberration test (ExxonMobil, 1991), Chinese Hamster Ovary cells were exposed to the test material ( Hydrocarbons, C12-C16, n-alkanes, isoalkanes, cyclics, <2% aromatics) at concentrations of 3.13, 6.26, 9.35 and 12.5 µg/mL for 10-h harvest and 12.5, 25, 37.5, 50 and 75 µg/mL for 20-h harvest, for 7 and 17 h, without metabolic activation and 37.5, 93.8, 188, 281, 375, 563 and 750 µg/mL for 10 and 20-h harvest, for 2 h, with metabolic activation.
Positive controls (mitomycin C without metabolic activation and cyclophosphamide with metabolic activation) induced the appropriate response. As there was no evidence of chromosome aberration induced over background, the test material was not classified according to the criteria of CLP Regulation (1272/2008).
In vitro Gene Mutation study in Mammalian Cells
In a key study (American Petroleum Institute, 1984), the test material (hydrodesulfurized kerosene) was examined for mutagenic activity in the mouse lymphoma forward mutation assay in the absence and presence of a liver S9 fraction for metabolic activation. The test material did not induce significant increases in the mutant frequency at the TK locus in L5178Y mouse lymphoma cells. Treatments up to 37.5 nl/ml without activation and 62.5 nl/ml with activation were assayed and high toxicities were induced without inducing significant increases in the mutant frequency. It is concluded in this study that the test material is not a mutagenic agent with or without activation. Classification is not warranted under the new Regulation (EC) 1272/2008 on classification, labeling, and packaging of substances and mixtures (CLP).
In Vivo
In a key mouse lymphoma forward mutation assay, the test material (hydrodesulfurised kerosene) showed no mutagenic properties (American Petroleum Institute, 1984).
Justification for classification or non-classification
The negative results observed in read across in vitro and in vivo genotoxicity assays do not warrant the classification of isoeicosane as a genotoxin under the new Regulation (EC) 1272/2008 on classification, labeling and packaging of substances and mixtures (CLP).
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.