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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 281-468-5 | CAS number: 83950-14-5
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Genetic toxicity: in vitro
Administrative data
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 07 June 2017 to 24 July 2017
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 017
- Report date:
- 2017
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 490 (In Vitro Mammalian Cell Gene Mutation Tests Using the Thymidine Kinase Gene)
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- other: forward mutation assay in mammalian cells
Test material
- Reference substance name:
- 4-[2-[4-[benzylmethyl(ethyl)amino]phenyl]vinyl]-1-(2-hydroxyethyl)pyridinium acetate
- EC Number:
- 281-468-5
- EC Name:
- 4-[2-[4-[benzylmethyl(ethyl)amino]phenyl]vinyl]-1-(2-hydroxyethyl)pyridinium acetate
- Cas Number:
- 83950-14-5
- Molecular formula:
- C24H27N2O.C2H3O2
- IUPAC Name:
- 4-[2-[4-[benzylmethyl(ethyl)amino]phenyl]vinyl]-1-(2-hydroxyethyl)pyridinium acetate
- Test material form:
- liquid
Constituent 1
Method
- Target gene:
- TK
Species / strain
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- CELLS USED
- Source of cells: Eurofins Munich stock culture
- Cell cycle doubling time: 10-12 hours
- Modal number of chromosomes: near diploid karyotype (40 ± 2 chromosomes)
- Normal (negative control) cell cycle time: 10-12
- Cloning Efficiency: usually more than 50%.
MEDIA USED
- Type and identity of media : RPMI 1640 complete medium
- Properly maintained: yes, cleansed and stored over liquid nitrogen (after thawing subcultured three times per week.)
- Periodically checked for Mycoplasma contamination: yes
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 liver microsomal fraction was prepared at Eurofins Munich. Male Wistar rats were induced with phenobarbital (80 mg/kg bw) and ß-naphthoflavone (100 mg/kg bw) for three consecutive days by oral route.
- Test concentrations with justification for top dose:
- without metabolic activation: 10, 25, 55, 60, 65, 70 and 90 µg/mL
with metabolic activation: 25, 40, 60, 70, 75, 80 and 90 µg/mL
Concentrations based on toxicity pre-test showing reduction of suspension growth at 50 µg/mL (RSG <10%) without metabolic activation and 150 µg/mL (RSG < 10%) with metabolic activation - Vehicle / solvent:
- 1% DMSO v/v
Controls
- Untreated negative controls:
- yes
- Remarks:
- treatment medium
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- benzo(a)pyrene
- ethylmethanesulphonate
- methylmethanesulfonate
- Remarks:
- for the positive controls the number of mutants was > GEF (126)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
- Cell density at seeding for cloning efficiency 1.6 cell/well
- Cell density at seeding for mutant determination: 2000 cells on 200 uL/well
DURATION
- Exposure time: 4 hours with and without metabolic activation in 11 mL RPMI medium with 5% horse serum (25 cm2 flasks) (10E7 cells)
- Expression time (cells in growth medium): 2 days at 37 °C in 5% CO2/95% humidified air in RPMI medium
- Selection time: incubationwith TFT during 12 days at 37 °C in 5% CO2/95% humidified air
NUMBER OF REPLICATIONS: 2 for cloning efficiency; 4 for mutagenicity
DETERMINATION OF CYTOTOXICITY
- Method: relative suspension growth/relative total growth - Rationale for test conditions:
- accoding to OECD 490
- Evaluation criteria:
- Positive (mutagenetic) result:
- The induced mutant frequency meets or exceeds the Global Evaluation factor (GEF) of 126 mutants per 106 cells and
- dose-dependent increase in mutant frequency is detected.
Besides, combined with a positive effect in the mutant frequency, an increased occurrence of small colonies (≥40% of total colonies) is an indication for potential clastogenic effects and/or chromosomal aberrations.
According to the OECD guideline, the biological relevance is considered first for the interpretation of results. Statistical methods might be used as an aid in evaluation of the test result.
A test item is considered to be negative if the induced mutant frequency is below the GEF and the trend of the test is negative.
Results and discussion
Test results
- Key result
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- RTG 11.5% at 90 ug/mL without metabolic activation and 11% at 90 ug/mL with metabolic activation
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks:
- MF [mutants / 106 cells] > 600; % small colonies > 40%
Any other information on results incl. tables
Main Experiment without metabolic activation
|
Test Group |
Conc. |
RCEa[%] |
RTGb[%] |
MFc[mutants/ 106cells] |
IMFd[mutants/ 106cells] |
GEFeexceeded |
Statistical Significant Increasef |
Precipitate |
[µg/mL] |
|||||||||
Exp without S9
|
C1 |
0 |
90.0 |
99.1 |
69.7 |
/ |
/ |
/ |
- |
C2 |
95.8 |
110.4 |
/ |
/ |
/ |
- |
|||
S1 |
0 |
100.0 |
100.0 |
57.2 |
/ |
/ |
/ |
- |
|
S2 |
/ |
/ |
/ |
- |
|||||
2 |
10 |
95.8 |
96.9 |
50.4 |
-6.8 |
- |
- |
- |
|
3 |
25 |
109.4 |
90.2 |
45.4 |
-11.8 |
- |
- |
- |
|
6 |
55 |
98.9 |
55.0 |
67.7 |
10.6 |
- |
- |
- |
|
7 |
60 |
87.3 |
36.7 |
52.0 |
-5.2 |
- |
- |
- |
|
8 |
65 |
102.2 |
29.2 |
55.6 |
-1.5 |
- |
- |
- |
|
9 |
70 |
91.4 |
16.4 |
43.7 |
-13.5 |
- |
- |
- |
|
13 |
90 |
95.8 |
11.5 |
44.6 |
-12.6 |
- |
- |
- |
|
EMS |
300 |
82.2 |
82.2 |
634.7 |
577.6 |
+ |
+ |
- |
|
MMS |
10 |
61.6 |
60.1 |
676.6 |
619.4 |
+ |
+ |
- |
|
Main Experiment with metabolic activation |
|||||||||
Exp with S9
|
C1 |
0 |
88.7 |
93.0 |
64.7 |
/ |
/ |
/ |
- |
C2 |
97.4 |
100.4 |
/ |
/ |
/ |
- |
|||
S1 |
0 |
100.0 |
100.0 |
65.0 |
/ |
/ |
/ |
- |
|
S2 |
/ |
/ |
/ |
- |
|||||
3 |
25 |
104.0 |
89.4 |
37.6 |
-27.4 |
- |
- |
- |
|
4 |
40 |
104.0 |
66.2 |
52.7 |
-12.3 |
- |
- |
- |
|
7 |
60 |
81.1 |
38.6 |
91.5 |
26.5 |
- |
- |
- |
|
9 |
70 |
87.4 |
29.8 |
79.3 |
14.3 |
- |
+ |
- |
|
10 |
75 |
79.9 |
17.2 |
81.3 |
16.4 |
- |
- |
- |
|
11 |
80 |
100.6 |
19.6 |
56.2 |
-8.8 |
- |
- |
- |
|
13 |
90 |
76.5 |
11.0 |
87.3 |
22.3 |
- |
- |
- |
|
B[a]P |
2.5 |
72.2 |
48.5 |
955.9 |
890.9 |
+ |
+ |
- |
C: Negative Controls
S: Solvent Controls
a: Relative Cloning Efficiency, RCE = [(CEdose group/ CEof corresponding controls) x 100]
Cloning Efficiency, CE = ((-LN (((96 - (mean P1,P2)) / 96)) / 1.6) x 100)
b: Relative Total Growth, RTG = (RSG x RCE)/100
c: Mutant Frequency, MF = {-ln [negative cultures/total wells (selective medium)] / -ln [negative cultures/total wells (non selective medium)]}x800
d: Induced Mutant Frequency, IMF = mutant frequency sample – mean value mutant frequency corresponding controls
e: Global Evaluation Factor, GEF (126); +: GEF exceeded, -: GEF not exceeded
f: statistical
significant increase in mutant frequency compared to solvent controls
(Mann Whitney test , p<0.05).
+: significant; -not significant
EMS Ethylmethanesulfonate; MMS:Methylmethanesulfonate; B[a]P:Benzo[a]pyrene
Applicant's summary and conclusion
- Conclusions:
- The substance is considered to be non-mutagenic in the in vitro mammalian cell gene mutation assay (thymidine kinase locus) in mouse lymphoma L5178Y cells.
- Executive summary:
In an vitro mammalian cell gene mutation assay (thymidine kinase locus) in mouse lymphoma L5178Y cells according to OECD 490, cells were exposed to the substance at concentrations between 1 and 90 ug/mL for 4 hours (with and without metabolic activation). The concentrations were based on cytotoxicity (as RGF). No increase in mutant colonies compared to vehicle and medium controls was observed. Therefore it can be concluded that the substance is non-mutagenic in this assay.
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.