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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 939-273-4 | CAS number: -
- 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 cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- From April 19, 2012 to August 01, 2012
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: GLP complaince with international guidelines
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 012
- Report date:
- 2012
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Version / remarks:
- Results obtained in a repeated experiment
- Deviations:
- yes
- Remarks:
- Results obtained in a repeated experiment
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)
- Version / remarks:
- Results obtained in a repeated experiment
- Deviations:
- yes
- Remarks:
- Results obtained in a repeated experiment
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- in vitro mammalian chromosome aberration test
Test material
- Reference substance name:
- Paraffin waxes and Hydrocarbon waxes C14-17, chloro, sulfochlorinated, low sulphonated, saponified
- EC Number:
- 939-273-4
- Molecular formula:
- CnHxCly(SO3Na)z • n= 14-17 • z < 0,5 • y = 0 ÷ 2,7
- IUPAC Name:
- Paraffin waxes and Hydrocarbon waxes C14-17, chloro, sulfochlorinated, low sulphonated, saponified
- Test material form:
- liquid: viscous
Constituent 1
Method
Species / strain
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Details on mammalian cell type (if applicable):
- - Periodically checked for karyotype stability: yes
- Periodically checked for Mycoplasma contamination: yes, regular intervals. - Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 tissue homogenate
The rat S9 liver tissue fraction used for the present study had the following characteristics:
Lot Number : 2879
Inducing Agents: Phenobarbital – 5,6-Benzoflavone
Received from: MOLTOX, Molecular Toxicology, Inc.
Storage at RTC: Approximately -80°C
Strain : Sprague Dawley
Sex of donors: Male
Protein content: 42.73 mg/mL
S9 mix
The S9 liver tissue fraction will be prepared according to RTC standard procedures or will be obtained from an appropriate supplier (MOLTOX, Molecular Toxicology, Inc., USA). Induction of drug metabolising enzyme-levels is routinely performed using phenobarbitone and betanaphtoflavone (mixed induction). Induction with Aroclor 1254 will be performed if specifically requested by the Sponsor. Records pertaining to the preparation of the S9 tissue fraction are kept on file at RTC.
The mixture of S9 tissue fraction and cofactors (S9 mix) will be prepared in the following proportions:
S9 tissue fraction 3.0 ml
NADP (0.1M) 0.4 ml
G-6-P (0.1M) 0.5 ml
KCl (0.33M) 1.0 ml
MgCl2 (0.1M) 0.4 ml
Hepes (0.2M) 1.0 ml
H.B.S.S. 3.7 ml
10.0 ml
Treatment cultures:
Treatment medium without metabolic activation will be prepared for each test point in the following proportions:
Test item or control solution 0.05 ml
Supplemented Ham's F10 medium 4.95 ml
Treatment medium with metabolic activation will be prepared containing the test item and S9 mix in the following proportions:
Test item or control solution 0.05 ml
S9 mix 0.50 ml
Supplemented Ham's F10 medium 4.45 ml - Test concentrations with justification for top dose:
- Experiment No. S9 Treatment time(hours) Harvest time (hours) Dose level(µg/mL)
1 ± 3 20 625, 313 ,156 ,78.1 and 39.1
2 - 20 20 80.0, 40.0 and 20.0
The dose levels of 0.30 and 0.10 µg/mL were selected for the scoring of cultures treated with Mitomycin-C (first and second experiment, respectively). The dose level of 15.0 µg/mL was selected for the scoring of cultures treated with Cyclophosphamide. - Vehicle / solvent:
- distilled water, culture medium, DMSO, ethanol, acetone.
Controlsopen allclose all
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- Remarks:
- with S9
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- mitomycin C
- Remarks:
- without S9
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
The medium used for the growth of the cells has the following composition:
Ham's F.10 499.0 ml
Streptomycin sulphate 50,000 IU/ml
Penicillin G 50,000 IU/ml 1.0 ml
Newborn Calf Serum 88.2 ml
DURATION
- Expression time (cells in growth medium): Each 25 sq.cm. flask will be seeded with 300,000 cells in supplemented Ham's F10. These cells should be in exponential growth phase at the time of treatment.
NUMBER OF REPLICATIONS: 2
NUMBER OF CELLS EVALUATED: 1 x 10^6 cells/ml
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index - Evaluation criteria:
- In this assay, the test item is considered to have clastogenic properties if the following criteria are all fulfilled:
(i) Statistically significant increases in the incidence of cells bearing aberrations are observed at any dose level over the concurrent control.
(ii) The increases must exceed the historical control values.
(iii) The increases are reproduced in both replicate cultures.
The evaluation is based on the set of results, which excludes gaps. A more detailed explanation of the criteria for evaluation of the results is given in the Study Protocol. - Statistics:
- For the statistical analysis, Fisher's Exact Test is used to compare the number of cells bearing aberrations (assumed to be Poisson distributed) in control and treated cultures. The analysis is performed using sets of data either including or excluding gaps. Following treatment in the absence of S9 metabolism, a statistically significant increase in the incidence of cells bearing structural aberrations including and excluding gaps was observed in the first main experiment, at the intermediate dose level selected for scoring. No other statistically significant increases were observed in any experiment, at any dose level, in the presence or absence of S9 metabolic activation. A statistically significant increase of endoreduplicated cells was observed at the highest dose level after treatment in the presence of S9 metabolic activation
Results and discussion
Test results
- 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:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: and osmolality: Following treatment with the test item, no remarkable variation of pH or osmolality was observed at any dose level, in the absence or presence of S9 metabolism. These results are not presented in this report but are retained in the study file and archived as indicated in the Study Protocol.
Any other information on results incl. tables
Deviations from Study Protocol
Results presented for the first experiment in the presence of S9 metabolic activation were obtained in a repeated experiment. In the original one, a low cellular growth was observed, probably due to the toxicity of S9 metabolic fraction. Data concerning the previous experiment are not presented in the report but will be archived as indicated in the Study Protocol.
On the basis of the above mentioned results and in accordance with the criteria for outcome of the study, the test item was not considered to induce structural chromosomal aberrations in Chinese hamster ovary cells in vitro.However, treatment with the test item in the presence of S9 metabolic activation induced a biologically relevant increase in the number of cells with endoreduplicated chromosomes, indicating that the test item may have the potential to inhibit cell cycle progression. Statistically significant increases in aberrant cells, compared with the relevant control values, were seen in cultures treated with the positive controls Mitomycin-C and Cyclophosphamide, indicating the correct functioning of the assay system.
Applicant's summary and conclusion
- Conclusions:
- On the basis of these results, it is concluded that PARAFFIN WAXES AND HYDROCARBON WAXES, CHLORO, SULFOCHLORINATED SAPONIFIED (C14-C17) SSP-SAMPLE 1 does not induce chromosomal aberrations in Chinese hamster ovary cells after in vitro treatment, under the reported experimental conditions.
It is also concluded that the test item induces endoreduplication in the presence of S9 metabolic activation. - Executive summary:
The test item PARAFFIN WAXES AND HYDROCARBON WAXES, CHLORO, SULFOCHLORINATED SAPONIFIED (C14-C17) SSP-SAMPLE 1 was assayed for the ability to cause chromosomal damage in Chinese hamster ovary cells, following in vitro treatment in the absence and presence of S9 metabolic activation. Two main experiments for chromosomal damage were performed.
In the first experiment, the cells were treated for 3 hours in the presence and absence of S9 metabolism, respectively. The harvest time of 20 hours corresponding to approximately 1.5 cell cycle length was used. As negative results were obtained, a second experiment was performed in the absence of S9 metabolism using the same harvest time. A continuous treatment until harvest at 20 hours was used.
Solutions of the test item were prepared in dimethylsulfoxide (DMSO). For the first main experiment, the maximum dose level for treatment was selected in agreement with the Study Protocol and on the basis of the solubility of the test item. Dose levels of 1250, 625, 313, 156, 78.1, 39.1,19.5 and 9.77 μg/mL were used both in the absence and presence of S9 metabolism.
The dose range used in the second experiment was modified to take into account the results observed in the previous experiment.
Dose levels of 160, 80.0, 40.0, 20.0, 10.0, 5.00, 2.50 and 1.25 μg/mL were used. Each experiment included appropriate negative and positive controls. Two cell cultures were prepared at each test point.
Dose levels were selected for the scoring of chromosomal aberrations on the basis of the cytotoxicity of the test item treatments as determined by the reduction of population doubling or relative cell count. Based on the obtained results, the following dose levels were selected for scoring:
Experiment No.
S9
Treatment time
(hours)
Harvest time (hours)
Dose level
(ug/mL)
1
+
-
3
20
625,313 and 156
156, 78.1 and 39.1
2
-
20
20
80.0, 40.0 and 20.0
One hundred metaphase spreads were scored for chromosomal aberrations from each culture, with the exception of one culture treated with Mitomycin-C from the first main experiment, where, due to the high incidence of aberrant cells (excluding gaps), scoring was terminated at 50 metaphases. Following treatment with test item, no statistically significant increase in the number of cells bearing structural aberrations, including or excluding gaps, was observed at any dose level or treatment time in the absence or presence of S9 metabolic activation. For the first main experiment, a statistically significant increase of endoreduplicated cells over the concurrent negative control was observed at the highest dose level selected for scoring (625 μg/mL) in the presence of S9 metabolism. Few endoreplicated cells were also observed at the intermediate dose level selected for scoring (313 μg/mL).
Statistically significant increases in the number of cells bearing aberrations (including and excluding gaps) were observed following treatment with the positive controls Cyclophosphamide and Mitomycin-C, indicating the correct functioning of the test system. It is concluded that PARAFFIN WAXES AND HYDROCARBON WAXES, CHLORO, SULFOCHLORINATED SAPONIFIED (C14-C17) SSP-SAMPLE 1 does not induce structural chromosomal aberrations in Chinese hamster ovary cells after in vitro treatment, under the reported experimental conditions. However, treatment with the test item induces an increase in the number of cells with endoreduplicated chromosomes only in the presence of metabolic activation and at the highest dose level selected for scoring (625 μg/mL), indicating that the test item may have the potential to inhibit cell cycle progression.
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.