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EC number: 825-403-6 | CAS number: 2060541-51-5
- 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
![](https://chesar.echa.europa.eu/o/diss-blank-theme/images/factsheets/A-REACH/factsheet/print_toxicological-information.png)
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
The key in vitro genotoxicity studies for this endpoint addressing both bacterial mutagenicity and mammalian mutagenicity were all negative within the whole range of the amphoteric, glycinate category. In order to strengthen the read across within the substance group, the genotoxic profile of all four substances were evaluated using the ToxTracker screening tool. ToxTracker is a panel of mammalian stem cell lines that contain different fluorescent reporters for induction of DNA damage, oxidative stress and protein damage. The differential induction of the GFP reporters as well as cytotoxicity of the tested compounds is determined by flow cytometry. No genotoxic or oxidative stress-inducing properties were detected for the amphoteric glycinate substances in this screening test. Also the QSAR toolbox, ACD/ToxSuite model, TOPKAT model, Derek tool and VEGA platform give no indications on mutagenic or carcinogenic properties for the data showing results with valid reliability index.
The only available in vitro mammalian clastogenicity test (OECD 473) is performed on the smallest substance within the group (shortest alky chain, lowest number of amine residues and carboxymethylated groups). The test report on the Human Lymphocyte Cytogenetics study conclude that Coco iminodiglycinate, (Amines, N-C12-18-alkyltrimethylenedi-, reaction products with chloroacetic acid, sodium salts with CAS no 2098351-38-1) is clastogenic based on a statistically significant increase on the mean percent aberrant cells at the top dose in Experiment II, 24 hour treatment without S9 metabolic activation. There was also a larger increase seen in Experiment I at 50µg/mL after 4 hour exposure without metabolic activation. However in this case the result was not statistically significant as the control value was at the top of the historical range. Also there was marked cytotoxicity with the relative mitotic index of 47% and the report concluded that the increase was cytotoxicity related.
The statistical significance seen in Experiment II at 50µg/mL was due to the control mean percent aberrant cells being at the bottom of the historical control range at 0.3% a factor of ten lower than at 4 hours. There was also cytotoxicity, but to a lesser degree than seen at 4 hours, with the relative mitotic index 58%. The higher relative mitotic index corresponded to a lower mean percentage of aberrant cells at 4.7% which is only just above the top of the historical control range of 4.2%. This compares to the mean percentage of 5.3% after 4 hours of treatment without S9 metabolic activation. Overall the increase in mean percent of aberrant cells seen in Experiment II at 50µg/mL can be considered most likely due to cytotoxicity rather than clastogenicity. Also there were clearly no effects on the mean percent aberrant cells even in Experiment I with 4 hour exposure in the presence of S9 metabolic activation at 100µg/mL in the absence of significant cytotoxicity. The lack of effects with metabolic activation indicates that there is little possibility of a systemic clastogenic effect in experimental animals.
As part of the weight of evidence there is also data for related substance Sodium cocopropylenediamine propionate (β-Alanine, N-(2-carboxyethyl)-N-[3-[(2-carboxyethyl)amino]propyl]-, N-C12-18-alkyl derivs., trisodium salts with CAS no 2136366-30-6) which has been tested in the in-vitro micronucleus assay also in human lymphocytes. This substance is closely structurally related to Coco iminodiglycinate, (Amines, N-C12-18 -alkyltrimethylenedi-, reaction products with chloroacetic acid, sodium salts with CAS no 2098351-38-1), the difference being that it contains propyl side chain groups rather than ethyl groups, but is otherwise the same being based on the same diamine. While it has not been included in the group for other end points there is no reason to believe that this difference in structure could have any influence on possible clastogenic potential. In the in-vitro micronucleus test on(β-Alanine, N-(2-carboxyethyl)-N-[3-[(2-carboxyethyl)amino]propyl]-, N-C12-18-alkyl derivs., trisodium salts with CAS no 2136366-30-6)there was no evidence of clastogenic or aneugenic activity at concentrations of 450µg/mL after 3 hour exposure without S9 metabolic activation, 600 µg/mL after 3 hour exposure with S9 metabolic activation and 200 µg/mL after 24 hour exposure without S9 metabolic activation.
Coco iminodiglycinate, (Amines, N-C12-18-alkyltrimethylenedi-, reaction products with chloroacetic acid, sodium salts with CAS no 2098351-38-1) has also been tested in the L5178Y TK +/- Mouse Lymphoma Assay. No clastogenic effect was induced by the test item, neither with nor without S9 metabolic activation, at concentrations up to 0.025mg/mL without S9 metabolic activation and 0.1mg/mL with S9 metabolic activation. None of the concentrations, with or without S9 metabolic activation, induced a frequency of small colonies >40%. Evidently the substance did not induce any clastogenic effect). The highest concentrations resulted in reduction of the relative total growth, with values of 11.1% without S9 metabolic activation and 17.4% with S9 metabolic activation, so they were cytotoxic levels.
The lack of any clastogenicity or anugenicity for the structurally related (β-Alanine, N-(2-carboxyethyl)-N-[3-[(2-carboxyethyl)amino]propyl]-, N-C12-18-alkyl derivs., trisodium salts with CAS no 2136366-30-6) at significantly higher concentrations in the in-vitro micronucleus assay together with a lack of any evidence of clastogenicity in theL5178Y TK +/- Mouse Lymphoma Assay with Coco iminodiglycinate, (Amines, N-C12-18-alkyltrimethylenedi-, reaction products with chloroacetic acid, sodium salts with CAS no 2098351-38-1), gives significant support to a weight of evidence assessment of clastogenicity. It supports a view that the apparent clastogenicity seen with Coco iminodiglycinate, (Amines, N-C12-18-alkyltrimethylenedi-, reaction products with chloroacetic acid, sodium salts with CAS no 2098351-38-1) in the in-vitro human lymphocyte chromosome aberrations study only after 24 hours exposure in the absence of S9 metabolic activation was not true clastogenicity but rather due to the cytotoxicity present at that concentration. The genotoxic profiles of all four substances have also been evaluated using the ToxTracker screening tool. ToxTracker is a panel of mammalian stem cell lines that contain different fluorescent reporters for induction of DNA damage, oxidative stress and protein damage. The differential induction of the GFP reporters as well as cytotoxicity of the tested compounds is determined by flow cytometry. No genotoxic or oxidative stress-inducing properties were detected for any of the amphoteric glycinate substances in this screening test. Also the QSAR toolbox, ACD/ToxSuite model, TOPKAT model, Derek tool and VEGA platform give no indications on mutagenic or carcinogenic properties for the data showing results with valid reliability index.
The overall conclusion is that Sodium cocoamphopolycarboxyglycinate (Amines, N-(3-aminopropyl)-N’-C12-18-alkyltrimethylenedi-, N-(carboxymethyl)derivs., sodium salts with CAS no 2060541-51-5)is not mutagenic, clastogenic or genotoxic.
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
- Study period:
- 3 - 20 September 1993.
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- This study was performed under GLP and according to OECD guideline from 1983. With this test it is not possible to identify certain oxidising mutagens, cross-linking agents and hydrazines. Such substances may be detected by E.coli WP2 strains or S. typhimurium TA102 which have an AT base pair at the primary reversion site in stead of GC base pairs which the strains tested in this study have.
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.5265 (The Salmonella typhimurium Bacterial Reverse Mutation Test)
- Principles of method if other than guideline:
- This study was performed according to OECD guideline from 1983. With this test it is not possible to identify certain oxidising mutagens, cross-linking agents and hydrazines. Such substances may be detected by E.coli WP2 strains or S. typhimurium TA102 which have an AT base pair at the primary reversion site in stead of GC base pairs which the strains tested in this study have.
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- mutation in the gene coding for histidine
- Species / strain / cell type:
- other: see below
- Details on mammalian cell type (if applicable):
- S. typhimurium TA 1535 hisG46 rfa uvrB
S. typhimurium TA 1537 hisC3076 rfa uvrB
S. typhimurium TA 1538 hisD3052 rfa uvrB
S. typhimurium TA 98 hisD3052 rfa uvrB pKM 10 1
S. typhimun'um TA 100 hisG46 rfa uvrB pKM101 - Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix
- Test concentrations with justification for top dose:
- 0, 50, 150, 500, 1500 and 5000 μg/plate
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: water
- Justification for choice of solvent/vehicle: As the test substance was a mixture of solids dissolved in water the solvent chosen for this assay was
water which was confirmed in a solubility test. - 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:
- Positive control substances:
without S9 mix:
Identity: N-Ethyl-N'-nitro-N-nitrosoguanidine
Supplier: Sigma
Batch: 67F-3700
Solvent: DMSO
Concentration: 5 microgram/plate for strain TA 1535
3 microgram/plate for strain TA 100
Identity: 9-Aminoacridine
Supplier: Sigma
Batch: 96F-05641
Solvent: DMSO
Concentration: 80 microgram/plate for strain TA 1537
Identity: 2-Aminoanthracene
Supplier: Aldrich
Batch: 61896
Solvent: DMSO
Concentration: 2 microgram/plate for strain TA 1538
1 microgram/plate for strain TA 98
With S9 mix:
dentity: 2-Aminoanthracene
Supplier: Aldrich
Batch: 0013406
Solvent: DMSO
Concentration: 2 microgram/plate for strain TA 1535 and TA 1537
0.5 microgram/plate for strain TA 1538 and TA 98
1 microgram/plate for strain TA 100
METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Preincubation period: none
- Exposure duration: 3 days
SELECTION AGENT (mutation assays): histidine defiencient agar
NUMBER OF REPLICATIONS: three petri dishes per dose level and an independent repeat was performed
NUMBER OF CELLS EVALUATED: all colonies were counted
DETERMINATION OF CYTOTOXICITY
- Method: assessment a background lawn
- Evaluation criteria:
- The mean number of revertant colonies for all treatment groups is compared with those obtained for solvent control groups. The mutagenic activity of a test substance is assessed by applying the following criteria:
(a) If treatment with a test substance produces an increase in revertant colony numbers of at least twice the concurrent solvent controls, with some evidence of a positive dose-relationship, in two separate experiments, with any bacterial strain either in the presence or absence of S-9 mix, it is considered to show evidence of mutagenic activity in this test system. No statistical analysis is performed.
(b) If treatment with a test substance does not produce reproducible increases of at least 1.5 times the concurrent solvent controls, at my dose level with any bacterial strain, it is considered to show no evidence of mutagenic activity in this test system. No statistical analysis is performed.
(c) If the results obtained fail to satisfy the criteria for a clear "positive" or "negative" response given in paragraphs (a) and (b), the following approach is taken in order to resolve the issue of the substance's mutagenic activity in this test system.
(i) Repeat tests may be performed using modifications of the experimental method. These modifications include (but are not restricted to), the use of a narrower dose range than that already tested; the use of different levels of liver homogenate S-9 fraction in the S-9 mix. Should an increase in revertant colony numbers be observed which satisfies paragraph (a) the substance is considered to show evidence of mutagenic activity in this .
test system. No statistical analysis is performed.
(ii) If no clear "positive" response can be obtained the test data may be subjected to analysis to determine the statistical significance of any observed increases in revertant colony numbers. The statisticd procedures used will be those described by Mahon et at. (1989). - Statistics:
- Not performed
- Species / strain:
- other: all strains tested
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- 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 examined
- Effects of osmolality: not examined
- Evaporation from medium: not examined
- Water solubility: sufficient for the test
- Precipitation: not observed
- Other confounding effects: not examined
RANGE-FINDING/SCREENING STUDIES: No cytotoxicty was observed in the preliminary toxicity test up to 5000 microgram/plate. this dose was therefore also selcted as the highest dose in the final study.
COMPARISON WITH HISTORICAL CONTROL DATA: not performed
No substantial increases in revertant colony numbers of any of the tester strains were observed following treatment with Ampholak 7CX at any dose level, in the presence or absence of S-9 mix, in either mutation test.
The concurrent positive control compounds demonstrated the sensitivity of the assay and the rnehbolising activity of the liver preparations. - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results: negative
It is concluded that, when tested in water, the test material (40% active ingredient and 60% water) was not mutagenic in this bacterial system. With this test it is not possible to identify certain oxidising mutagens, cross-linking agents and hydrazines. Such substances may be detected by E.coli WP2 strains or S. typhimurium TA102 which have an AT base pair at the primary reversion site in stead of GC base pairs which the strains tested in this study have. Under the given conditions, the active ingredient Sodium cocoamphopolycarboxyglycinate (Amines, N-(3-aminopropyl)-N’-C12-18-alkyltrimethylenedi-, N-(carboxymethyl)derivs., sodium salts with CAS no 2060541-51-5) is considered to be non mutagenic. - Executive summary:
In this in vitro assessment of the mutagenic potential of the test material, histidine dependent auxotrophic mutanrs of Salmonella typhimunum (strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100) were exposed to the test substance, diluted in water which was also used as a negative control. Two independent mutation tests were performed accoriding to OECD guodlien and under GLP, in the presence and absence of liver preparations from Aroclor 1254-induced rats. In the preliminary dose range finding study with dose levels of up to 5000 μg/plate no toxicity was observed. A top dose level of 5000 μg/plate was chosen for the subsequent mutation study. Other dose levels used in the mutation assays were: 1500, 500, 150, 50 μg/plate. No evidence of mutagenic activity was seen at any dose level of the test material in either mutation test. The concurrent positive control compounds demonstrated the sensitivity of the assay and the metabolising activity of the liver preparations. It is concluded that, when tested in water, the test materal was not mutagenic in this bacterial system. Under the given conditions, the active ingredient Sodium cocoamphopolycarboxyglycinate (Amines, N-(3-aminopropyl)-N’-C12-18-alkyltrimethylenedi-, N-(carboxymethyl)derivs., sodium salts with CAS no 2060541-51-5) is therefore considered to be non mutagenic. With this test it is not possible to identify certain oxidising mutagens, cross-linking agents and hydrazines. Such substances may be detected by E.coli WP2 strains or S. typhimurium TA102 which have an AT base pair at the primary reversion site in stead of GC base pairs which the strains tested in this study have.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- read-across based on grouping of substances (category approach)
- Adequacy of study:
- key study
- Justification for type of information:
- Read-across (Category)
The key in vitro genotoxicity studies for this endpoint addressing both bacterial mutagenicity and mammalian mutagenicity were all negative within the whole range of the amphoteric, glycinate category. In order to strengthen the read across within the substance group, the genotoxic profile of all four substances were evaluated using the ToxTracker screening tool. ToxTracker is a panel of mammalian stem cell lines that contain different fluorescent reporters for induction of DNA damage, oxidative stress and protein damage. The differential induction of the GFP reporters as well as cytotoxicity of the tested compounds is determined by flow cytometry. No genotoxic or oxidative stress-inducing properties were detected for the amphoteric glycinate substances in this screening test. Also the QSAR toolbox, ACD/ToxSuite model, TOPKAT model, Derek tool and VEGA platform give no indications on mutagenic or carcinogenic properties for the data showing results with valid reliability index. - Species / strain:
- lymphocytes: Human
- Metabolic activation:
- with and without
- Genotoxicity:
- positive
- Remarks:
- Different intepretation of the data if possible. See Applicant's summary and conclusion.
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Conclusions:
- The test report on the Human Lymphocyte Cytogenetics study conclude that Coco iminodiglycinate, (Amines, N-C12-18-alkyltrimethylenedi-, reaction products with chloroacetic acid, sodium salts with CAS no 2098351-38-1) is clastogenic based on a statistically significant increase on the mean percent aberrant cells at the top dose in Experiment II, 24 hour treatment without S9 metabolic activation. There was also a larger increase seen in Experiment I at 50µg/mL after 4 hour exposure without metabolic activation. However in this case it was not statistically significant as the control value was at the top of the historical range. Also there was marked cytotoxicity with the relative mitotic index of 47% and the report concluded that the increase was cytotoxicity related.
The statistical significance seen in Experiment II at 50µg/mL was due to the control mean percent aberrant cells being at the bottom of the historical control range at 0.3% a factor of ten lower than at 4 hours. There was also cytotoxicity, but to a lesser degree than seen at 4 hours, with the relative mitotic index 58%. The higher relative mitotic index corresponded to a lower mean percentage of aberrant cells at 4.7% which is only just above the top of the historical control range of 4.2%. This compares to the mean percentage of 5.3% after 4 hours of treatment without S9 metabolic activation. Overall the increase in mean percent of aberrant cells seen in Experiment II at 50µg/mL can be considered most likely due to cytotoxicity rather than clastogenicity. Also there were clearly no effects on the mean percent aberrant cells even in Experiment I with 4 hour exposure in the presence of S9 metabolic activation at 100µg/mL in the absence of significant cytotoxicity. The lack of effects with metabolic activation indicates that there is little possibility of a systemic clastogenic effect in experimental animals.
As part of the weight of evidence there is also data for related substance Sodium cocopropylenediamine propionate (β-Alanine, N-(2-carboxyethyl)-N-[3-[(2-carboxyethyl)amino]propyl]-, N-C12-18-alkyl derivs., trisodium salts with CAS no 2136366-30-6) which has been tested in the in-vitro micronucleus assay also in human lymphocytes. This substance is closely structurally related to Coco iminodiglycinate, (Amines, N-C12-18-alkyltrimethylenedi-, reaction products with chloroacetic acid, sodium salts with CAS no 2098351-38-1), the difference being that it contains propyl side chain groups rather than ethyl groups, but is otherwise the same being based on the same diamine. While it has not been included in the group for other end points there is no reason to believe that this difference in structure could have any influence on possible clastogenic potential. In the in-vitro micronucleus test on (β-Alanine, N-(2-carboxyethyl)-N-[3-[(2-carboxyethyl)amino]propyl]-, N-C12-18-alkyl derivs., trisodium salts with CAS no 2136366-30-6) there was no evidence of clastogenic or aneugenic activity at concentrations of 450µg/mL after 3 hour exposure without S9 metabolic activation, 600 µg/mL after 3 hour exposure with S9 metabolic activation and 200 µg/mL after 24 hour exposure without S9 metabolic activation.
Coco iminodiglycinate, (Amines, N-C12-18-alkyltrimethylenedi-, reaction products with chloroacetic acid, sodium salts with CAS no 2098351-38-1)has also been tested in the L5178Y TK +/- Mouse Lymphoma Assay. No clastogenic effect was induced by the test item, neither with nor without S9 metabolic activation, at concentrations up to 0.025mg/mL without S9 metabolic activation and 0.1mg/mL with S9 metabolic activation. None of the concentrations, with or without S9 metabolic activation, induced a frequency of small colonies >40%. Evidently the substance did not induce any clastogenic effect). The highest concentrations resulted in reduction of the relative total growth, with values of 11.1% without S9 metabolic activation and 17.4% with S9 metabolic activation, so they were cytotoxic levels.
The lack of any clastogenicity or anugenicity for the structurally related (β-Alanine, N-(2-carboxyethyl)-N-[3-[(2-carboxyethyl)amino]propyl]-, N-C12-18-alkyl derivs., trisodium salts with CAS no 2136366-30-6) at significantly higher concentrations in the in-vitro micronucleus assay together with a lack of any evidence of clastogenicity in the L5178Y TK +/- Mouse Lymphoma Assay with Coco iminodiglycinate, (Amines, N-C12-18-alkyltrimethylenedi-, reaction products with chloroacetic acid, sodium salts with CAS no 2098351-38-1), gives significant support to a weight of evidence assessment of clastogenicity. It supports a view that the apparent clastogenicity seen with Coco iminodiglycinate, (Amines, N-C12-18-alkyltrimethylenedi-, reaction products with chloroacetic acid, sodium salts with CAS no 2098351-38-1)in the in-vitro human lymphocyte chromosome aberrations study only after 24 hours exposure in the absence of S9 metabolic activation was not true clastogenicity but rather due to the cytotoxicity present at that concentration. The genotoxic profiles of all four substances have also been evaluated using the ToxTracker screening tool. ToxTracker is a panel of mammalian stem cell lines that contain different fluorescent reporters for induction of DNA damage, oxidative stress and protein damage. The differential induction of the GFP reporters as well as cytotoxicity of the tested compounds is determined by flow cytometry. No genotoxic or oxidative stress-inducing properties were detected for any of the amphoteric glycinate substances in this screening test. Also the QSAR toolbox, ACD/ToxSuite model, TOPKAT model, Derek tool and VEGA platform give no indications on mutagenic or carcinogenic properties for the data showing results with valid reliability index.
The overall conclusion is that Coco iminodiglycinate, (Amines, N-C12-18-alkyltrimethylenedi-, reaction products with chloroacetic acid, sodium salts with CAS no 2098351-38-1) is not clastogenic. - Executive summary:
A chromosome aberration assay was carried out in order to investigate a possible potential of Coco iminodiglycinate to induce structural chromosome aberrations in human lymphocytes.The metaphases were prepared 24 h after start of treatment with the test item. The treatment interval was 4 h without and with metabolic activation (experiment I) and 24 h without metabolic activation (experiment II). Duplicate cultures were set up. Per culture 150 metaphases were scored for structural chromosomal aberrations.
The test item was dissolved in cell culture medium and diluted prior to treatment.
The following concentrations were evaluated:
Experiment I
Without metabolic activation, 4 h treatment, 24 h preparation interval: 10, 25 and 50 µg/mL
With metabolic activation, 4 h treatment, 24 h preparation interval: 25, 50 and 100 µg/mL
Experiment II
Without metabolic activation, 24 h treatment, 24 h preparation interval: 10, 25 and 50 µg/mL
No precipitation of the test item was noted without and with metabolic activation in all dose groups evaluated in experiment I and II.
In experiment I without metabolic activation, toxic effects (decrease below 70% rel. mitotic index) were seen at a concentration of 50 µg/mL and higher. With metabolic activation, cytotoxicity was observed at a concentration of 250 µg/mL. In experiment II without metabolic activation, a biologically relevant decrease of the relative mitotic index was noted at the concentration of 50 µg/mL and higher.
No reduction of the proliferation index was indicated in experiment I with and without metabolic activation. In experiment II a biologically relevant decreases of the proliferation index was observed, thus indicating a cell cycle delay.
In experiment I without metabolic activation the aberration rates were within the historical negative control data, except for the highest concentration evaluated. This induction seemed to be cytotoxicity related as the mitotic index of the concentration 50 µg/mL was reduced to 47%. With metabolic activation, the aberration rates of the negative control and the concentration 25 µg/mL were slightly shifted to a higher degree of chromosomal aberrations regarding the historical control range of the negative control . Given that the higher test substance concentrations were within the control data, the marginal induction was not regarded as biologically relevant.
In experiment II without metabolic activation the aberration rates were within the historical negative control data, except for the highest concentration of 50 µg/mL.
In the experiments I and II without and withmetabolic activation no biologically relevant increase in the frequencies of polyploid cells was found after treatment with the test item as compared to the controls.
The Fisher´s exact test was performed to verify the results in the experiment. A statistical significance (p < 0.05) was only present at the concentration 50 µg/mL in experiment II.
The chi² Test for trend was performed to test whether there was a concentration-related increase in chromosomal aberrations. No statistically significant increase was observed in experiment I without and with metabolic activation. In experiment II, a dose-related increase in chromosome damage was determined.
EMS (400 and 600 µg/mL) and CPA (5.0 µg/mL) were used as positive controls and induced distinct and biologically relevant increases in cells with structural chromosomal aberrations, thus proving the efficiency of the test system to indicate potential clastogenic effects.
There was evidence of chromosome aberration induced over background.
This study is classified as acceptable. This study satisfies the requirement for Test Guideline OECD 473 for in vitrocytogenetic mutagenicity data.
- Endpoint:
- in vitro gene mutation study in bacteria
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- read-across based on grouping of substances (category approach)
- Adequacy of study:
- key study
- Justification for type of information:
- Read-across (Category)
The key in vitro genotoxicity studies for this endpoint addressing both bacterial mutagenicity and mammalian mutagenicity were all negative within the whole range of the amphoteric, glycinate category. In order to strengthen the read across within the substance group, the genotoxic profile of all four substances were evaluated using the ToxTracker screening tool. ToxTracker is a panel of mammalian stem cell lines that contain different fluorescent reporters for induction of DNA damage, oxidative stress and protein damage. The differential induction of the GFP reporters as well as cytotoxicity of the tested compounds is determined by flow cytometry. No genotoxic or oxidative stress-inducing properties were detected for the amphoteric glycinate substances in this screening test. Also the QSAR toolbox, ACD/ToxSuite model, TOPKAT model, Derek tool and VEGA platform give no indications on mutagenic or carcinogenic properties for the data showing results with valid reliability index. - Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- experiment I and II
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- experiment I and II
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- experiment I and II
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- experiment I and II
- Species / strain:
- S. typhimurium TA 102
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- experiment I and II
- Conclusions:
- Interpretation of results (migrated information): negative
In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, Coco iminodiglycinate (Amines, N-C12-18-alkyltrimethylenedi-, reaction products with chloroacetic acid, sodium salts with CAS no 2098351-38-1) did not cause gene mutations by base pair changes or frameshifts in the genome of the tester strains used. Therefore, Coco iminodiglycinate is considered to be non-mutagenic in this bacterial reverse mutation assay. - Executive summary:
In a reverse gene mutation assay in bacteria, strainsTA 98, TA 100, TA 1535, TA 1537 and TA 102ofSalmonella typhimuriumwere exposed toCoco iminodiglycinateat concentrations of0.0100, 0.0316, 0.100, 0.316, 1.0, 2.5 and 5.0 µL/plate(experiment I and II), in the presence and absence of mammalian metabolic activation according to the plate incorporation method (experiment I) and the pre-incubation method (experiment II).Coco iminodiglycinate (Amines, N-C12-18-alkyltrimethylenedi-, reaction products with chloroacetic acid, sodium salts with CAS no 2098351-38-1)was tested up to the limit concentration of 5.0 µL/plate in all tester strains used.
The positive controls induced the appropriate responses in the corresponding strains.There was no evidence of induced mutant colonies over background.
This study is classified as acceptable. This study satisfies the requirement for Test Guideline OPPTS 870.5100; OECD 471 for in vitro mutagenicity (bacterial reverse gene mutation) data.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- read-across based on grouping of substances (category approach)
- Adequacy of study:
- key study
- Justification for type of information:
- Read-across (Category)
The key in vitro genotoxicity studies for this endpoint addressing both bacterial mutagenicity and mammalian mutagenicity were all negative within the whole range of the amphoteric, glycinate category. In order to strengthen the read across within the substance group, the genotoxic profile of all four substances were evaluated using the ToxTracker screening tool. ToxTracker is a panel of mammalian stem cell lines that contain different fluorescent reporters for induction of DNA damage, oxidative stress and protein damage. The differential induction of the GFP reporters as well as cytotoxicity of the tested compounds is determined by flow cytometry. No genotoxic or oxidative stress-inducing properties were detected for the amphoteric glycinate substances in this screening test. Also the QSAR toolbox, ACD/ToxSuite model, TOPKAT model, Derek tool and VEGA platform give no indications on mutagenic or carcinogenic properties for the data showing results with valid reliability index. - Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Conclusions:
- Interpretation of results: negative
In conclusion, in the described mutagenicity test under the experimental conditions reported, the test item Coco iminodiglycinate, (Amines, N-C12-18-alkyltrimethylenedi-, reaction products with chloroacetic acid, sodium salts with CAS no 2098351-38-1) 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:
The test item Coco iminodiglycinate, (Amines, N-C12-18-alkyltrimethylenedi-, reaction products with chloroacetic acid, sodium salts with CAS no 2098351-38-1) was assessed for its potential to induce mutations at the mouse lymphoma thymidine kinase locus using the cell line L5178Y. The selection of the concentrations used in the main experiment was based on data from the pre-experiment. In the main experiment without metabolic activation 0.025 mg/mL and with metabolic activation 0.1 mg/mL were selected as the highest concentrations. The experiment without and with metabolic activation was performed as a 4 h short-term exposure assay.
The test item Coco iminodiglycinate, (Amines, N-C12-18-alkyltrimethylenedi-, reaction products with chloroacetic acid, sodium salts with CAS no 2098351-38-1) was investigated at the following concentrations:
without metabolic activation:
0.0005, 0.0010, 0.0025, 0.005, 0.01, 0.015, 0.02 and 0.025 mg/mL
and with metabolic activation:
0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09 and 0.1 mg/mL
No precipitation of the test item was noted in the experiment.
Growth inhibition was observed in the main experiment without and with metabolic activation.
In the main experiment without metabolic activation the relative total growth (RTG) was 11.1% for the highest concentration (0.025 mg/mL) evaluated. The highest concentration evaluated with metabolic activation was 0.1 mg/mL with a RTG of 17.4%. In the main experiment no biologically relevant increase of mutants was found after treatment with the test item (without and with metabolic activation). The Global Evaluation Factor (GEF; defined as the mean of the negative/vehicle mutant frequency plus one standard deviation; data gathered from ten laboratories) was not exceeded by the induced mutant frequency at any concentration.
No dose-response relationship was observed. Additionally, in the main experiment colony sizing showed no clastogenic effects induced by the test item under the experimental conditions (without and with metabolic activation).
EMS, MMS and B[a]P were used as positive controls and showed distinct and biologically relevant effects in mutation frequency. Additionally, MMS and B[a]P significantly increased the number of small colonies, thus proving the efficiency of the test system to indicate potential clastogenic effects.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Remarks:
- ToxTracker assay
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- Between 11/08/2016 and 17/08/2016
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- study well documented, meets generally accepted scientific principles, acceptable for assessment
- Justification for type of information:
- ToxTracker is a panel of mammalian stem cell lines that contain different fluorescent reporters for induction of DNA damage, oxidative stress and protein damage. The differential induction of the GFP reporters as well as cytotoxicity of the tested compounds is determined by flow cytometry. The selected compounds were analysed in the absence and presence of an S9 rat liver extract-based metabolising system. Exposure to control compounds is included in each test to determine technical performance and reproducibility of the ToxTracker assay. Quantitative data analysis is done using ToxPlot software.
The study was not performed under GLP, however general principles to conduct proper scientifically correct in vitro experiments were adhered to. - Qualifier:
- no guideline available
- Principles of method if other than guideline:
- The ToxTracker assay is a panel of six validated GFP-based mouse embryonic stem (mES) reporter cell lines that can be used to identify the biological reactivity and potential carcinogenic properties of newly developed compounds in a single test. ToxTracker is a mammalian stem cell-based assay that monitors activation of specific cellular signalling pathways for detection of the biological reactivity of compounds. In contrast to the cancer-derived cell lines that are currently used for in vitro genotoxicity testing, stem cells are genetically stable and proficient in all cellular pathways required for accurate detection of potentially carcinogenic properties of compounds. Extensive whole-genome transcription profiling has led to identification of a panel of biomarker genes that are preferentially activated upon exposure to different classes of carcinogens and toxicants. (1) To allow easy assessment of the activation status of these biomarker genes, the system uses generated green fluorescent (GFP) mES reporter cell lines. These reporters were created using bacterial artificial chromosomes (BAC) that contain the complete biomarker gene including promoter and regulatory elements ensuring physiological regulation of the GFP reporters following transfection into stem cells. (2)
ToxTracker consists of a panel of six different mES GFP reporter cell lines representing four distinct biological responses that are associated with carcinogenesis, i.e. general cellular stress, DNA damage, oxidative stress and the unfolded protein response. (3)
(1) Hendriks, G., Atallah, M., Raamsman, M., Morolli, B., van der Putten, H., Jaadar, H., Tijdens, I., Esveldt-van Lange, R., Mullenders, L., van de Water, B., et al. (2011). Sensitive DsRed fluorescence-based reporter cell systems for genotoxicity and oxidative stress assessment. Mutat Res 709-710, 49–59.
(2) Hendriks, G., Atallah, M., Morolli, B., Calléja, F., Ras-Verloop, N., Huijskens, I., Raamsman, M., van de Water, B., and Vrieling, H. (2012). The ToxTracker assay: novel GFP reporter systems that provide mechanistic insight into the genotoxic properties of chemicals. Toxicol. Sci. 125, 285–298.
(3) Hendriks G, Derr RS, Misovic B, Morolli B, Calléja FMGR, Vrieling H. (2016). The Extended ToxTracker Assay Discriminates Between Induction of DNA Damage, Oxidative Stress, and Protein Misfolding. Toxicol Sci. 150, 190–203. - GLP compliance:
- no
- Remarks:
- This screening study was not performed under GLP, however general principles to conduct proper scientifically correct in vitro experiments were adhered to.
- Type of assay:
- other: ToxTracker assay
- Target gene:
- Biological damage: Cellular pathway: Biomarker gene:
DNA damage ATR/Chk1 DNA damage signaling Bscl2
NF-kB signaling Rtkn
Oxidative stress Nrf2 antioxidant response Srxn1
Nrf2-independent Blvrb
Protein damage Unfolded protein response Ddit3
Cellular stress p53 signaling Btg2 - Species / strain / cell type:
- mammalian cell line, other:
- Remarks:
- GFP-based mouse embryonic stem (mES) reporter cell line
- Details on mammalian cell type (if applicable):
- Extensive whole-genome transcription profiling has led to identification of a panel of biomarker genes that are preferentially activated upon exposure to different
classes of carcinogens and toxicants. (1) To allow easy assessment of the activation status of these biomarker genes, green fluorescent (GFP) mES reporter cell lines have been generated. These reporters were created using bacterial artificial chromosomes (BAC) that contain the complete biomarker gene including promoter and regulatory elements ensuring physiological regulation of the GFP reporters following transfection into stem cells. (2)
(1) Hendriks, G., Atallah, M., Raamsman, M., Morolli, B., van der Putten, H., Jaadar, H., Tijdens, I., Esveldt-van Lange, R., Mullenders, L., van de Water, B., et al. (2011). Sensitive DsRed fluorescence-based reporter cell systems for genotoxicity and oxidative stress assessment. Mutat Res 709-710, 49–59.
(2) Hendriks, G., Atallah, M., Morolli, B., Calléja, F., Ras-Verloop, N., Huijskens, I., Raamsman, M., van de Water, B., and Vrieling, H. (2012). The ToxTracker assay: novel GFP reporter systems that provide mechanistic insight into the genotoxic properties of chemicals. Toxicol. Sci. 125, 285–298. - Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 liver extract from alachlor-induced rats (Moltox)
- Test concentrations with justification for top dose:
- The highest compound concentration will induce significant cytotoxicity (50-70%). The appropriate dose range for the ToxTracker analysis is determined in a extensive dose range finding. For the dose range finding, as well as the ToxTracker analysis a maximum concentration of 10 mM or 2 mg/ml is used, in line with the advised maximum concentration in the current guidelines for in vitro genotoxicity testing.
Doses used for testing were 0, 0.004, 0.008, 0.016, 0.031, 0.063% of Amines, N-(3-aminopropyl)-N’-C12-18-alkyltrimethylenedi-, N-(carboxymethyl)derivs., sodium salts (CAS no 2060541-51-5) without S9.
Doses used for testing were 0, 0.008, 0.016, 0.031, 0.063, 0.125% of Amines, N-(3-aminopropyl)-N’-C12-18-alkyltrimethylenedi-, N-(carboxymethyl)derivs., sodium salts (CAS no 2060541-51-5) with S9. - Vehicle / solvent:
- DMSO
- Untreated negative controls:
- yes
- Remarks:
- DMSO as solvent
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- Positive controls:
- yes
- Positive control substance:
- other:
- Remarks:
- Without S9: Cisplatin = positive control DNA damage. Dimethyl maleate = positive control oxidative stress. Tunicamycin = positive control protein damage. With S9: Cisplatin = positive control DNA damage Aflatoxin = positive control metabolic activation
- Details on test system and experimental conditions:
- The ToxTracker assay requires only standard cell culture facilities and a low-end flow cytometer. The ToxTracker reporter cells are maintained by culturing them in gelatin-coated dishes in the presence of irradiated primary mouse embryonic fibroblasts (MEFs). During chemical exposures and reporter analysis the ToxTracker cells are cultured in the absence of fibroblasts in Buffalo rat liver (BRL)-conditioned mES cell culture medium.
For chemical testing, the six independent mES reporter cell lines are seeded in gelatin-coated 96-well cell culture plates in 200 μl BRL-conditioned ES cell medium (40.000 cells per well). 24 h after seeding the cells in the 96-well plates, medium is aspirated and fresh BRL-conditioned ES cell medium containing 10% foetal calf serum and the diluted chemicals is added to the cells. For each tested compound, five concentrations are tested in 2-fold dilutions. The highest compound concentration will induce significant cytotoxicity (50-70%). The appropriate dose range for the ToxTracker analysis is determined in a extensive dose range finding. For the dose range finding, as well as the ToxTracker analysis a maximum concentration of 10 mM or 2 mg/ml is used, in line with the advised maximum concentration in the current guidelines for in vitro genotoxicity testing. In case concentrations are limited by solubility or the occurrence of precipitation in the culture medium, the maximum soluble concentration will be used in the assay.
For the dose range finding, wild type mES cells are exposed to 20 different concentrations of the test substances. Cytotoxicity is estimated by cell count after 24 h exposure using a flow cytometer and is expressed as percentage of viable cells after 24 h. exposure compared to unexposed controls. Compounds are diluted in DMSO, water or PBS (see Table 3). For cytotoxicity assessment in the ToxTracker assay, the relative cell survival for the six different reporter cell lines is averaged.
GFP reporter induction is always compared to a vehicle control treatment. DMSO concentration is similar in all wells for a particular compound and never exceeds 1%. All compounds have been tested in at least three completely independent experiments. Positive reference treatments with cisplatin (DNA damage), diethyl maleate (oxidative stress) and tunicamycin (unfolded protein response) are included in all experiments. Metabolic activation is included in the ToxTracker assay by addition of S9 liver extract from alachlor-induced rats (Moltox). Cells are exposed to five concentrations of the test compounds in the presence of S9 and required co-factors (RegenSysA+B, Moltox) for 3 h. After washing, cells are incubated for 24 h in fresh BRLconditioned ES cell medium.
Induction of the GFP reporters is determined after 24 h exposure using a Guava easyCyte 8HT flow cytometer (Millipore). Only GFP expression in intact single cells is determined. Mean GFP fluorescence and cell concentrations in each well is measured, which is used for cytotoxicity assessment. In case auto-fluorescence of the test substances was observed in the dose range finding, wild type mES cells were exposed to the samples at the same concentrations as used in the ToxTracker. The mean fluorescence caused by the compound was then subtracted from the ToxTracker results of the respective compound. - Evaluation criteria:
- Induction of the six reporter genes after treatment with test substance up to cytotoxic concentrations.
Validity of the assay was confirmed using exposure to the reference compounds specific for the pathways evaluated. Diethyl maleate (DEM) induced primarily the oxidative stress related reporters Srxn1 and Blvrb, tunicamycin induced the unfolded/misfolded protein stress response (Ddit3), and cisplatin showed induction of the DNA damage response (Bscl2, Rtkn) and p53-mediated cellular stress (Btg2).
Activation of the different GFP reporters by the control compounds is fully compliant with historical data thereby confirming the technical validity of the performed tests. - Statistics:
- A statistical evaluation of the results is not regarded as necessary.
- Species / strain:
- other: mammalian cell line
- Remarks:
- GFP-based mouse embryonic stem (mES) reporter cell line
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- experiemnt I and II (with and without S9)
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- All four tested substances within the ampoteric, glycinate substance group induced significant levels of cytotoxicity (>50%) at a concentrations of 0.02% or lower when tested in the absence of S9 liver extract. None of the four tested substances showed increased cytotoxicity when cells were exposed in presence of S9 rat liver extract, indication that the substances did not require metabolic activation to become biological reactive. The 50% cytotoxicity level for the tested substances in presence of S9 varied between 0.04 and 0.07%. The reduced cytotoxicity when tested in the presence of S9 which is likely caused by a reduced exposure time of 3 h compared to 24 h for exposures in absence of S9. The six ToxTracker reporter cell lines showed a comparable cytotoxic response to the test compounds. For this reason, the cell survival graphs in the GFP induction figures show the average cytotoxicity of the six different cell lines.
None of the four tested ampoteric, glycinate substances induced the Bscl2-GFP or Rtkn-GFP DNA damage reporters when tested in the absence of a metabolising system. Activation of the Bscl2-GFP reporter is associated with induction of promutagenic DNA lesions that interfered with DNA replication. Activation of the Rtkn-GFP genotoxicity reporters correlates with induction of DNA strand breaks. Also when tested in the presence of a metabolising system, non of the ampoteric, glycinate substances activated the genotoxicity reporters in the ToxTracker assay. Together, no indication for genotoxic properties was found for the four tested ampoteric, glycinate substances.
None of the four tested substances induced significant levels of cellular oxidative stress when exposed in absence of a metabolising system, even at high cytotoxic concentrations. Oxidative stress is established by the Srxn1- GFP and Blvrb-GFP reporters. When tested in the presence of S9, only the biggest substance (longest alkyl-unsaturated, highest number amine and carboxyl groups) with CAS no 2060541-49-1 slightly activated the Srxn1-GFP oxidative stress reporter. Srxn1 activation is directly controlled by the Nrf2 anti-oxidant response pathway. However, the induction levels were >10-fold lower compared to the positive control Diethyl maleate. No oxidative stress by the other three substances was detected when exposed in the presence of S9.
In the absence of a metabolising system, all the amphoteic, glysincate substances but the biggest one with CAS no 2060541-49-1 significantly induced the Ddit3-GFP reporter, that is associated with protein damage and the unfolded protein response. Activation of the unfolded protein response was already observed at low cytotoxic concentrations. When tested in the presence of a metabolising system, induction of protein damage was reduced, which is generally correlated to the S9 exposure Protocol. After the 3 h exposure of cells in presence of compound and S9, cells were washed and allowed to recover up to 24 h. During this recovery time, cells are able to repair the cellular damage and will down-regulate the unfolded protein response. Only the smallest amphoteric, glycinate substance with CAS no 2098351-38-1 also activated the protein damage response in the presence of S9.
All criteria of validity were met. - Remarks on result:
- other:
- Remarks:
- No genotoxicity nor indiction of oxidative stress was observed in the ToxTracker assay in any of the tested concentration in the absence or presence of a metabolising system.
- Conclusions:
- Interpretation of results: negative
In conclusion, it can be stated that in the described genotoxicity test and under the experimental conditions reported, the following can be concluded for Sodium cocoamphopolycarboxyglycinate (CAS nr 2060541-51-5):
• Strong cytotoxicity (>50%) was observed at a concentration of 0.01%
• No genotoxicity was observed in the ToxTracker assay at the highest tested concentration in the absence or presence of a metabolising system.
• No induction of oxidative stress was observed in the ToxTracker assay at the highest tested concentration in the absence or presence of a metabolising system.
• Strong induction of the unfolded protein response was observed, to comparable levels as the positive control.
• No increased reactivity was observed in the presence of a metabolising system.
Therefore, Sodium cocoamphopolycarboxyglycinate (CAS nr 2060541-51-5) is considered to be non-mutagenic in this ToxTracker screening test. - Executive summary:
ToxTracker is a screening study using a panel of mammalian stem cell lines that contain different fluorescent reporters for induction of DNA damage, oxidative stress and protein damage. All four substances within the amphoteric, glycinate substance group were tested in this assay up to cytotoxic levels with and without S9 rat liver extract-based metabolising system. The positive controls induced the appropriate responses. No genotoxic or oxidative stress-inducing properties were detected for any of the amphoteric, glycinate substances. All but the biggest substance (longest alkyl-unsaturated, highest number amine and carboxyl groups) with CAS no 2060541-49-1 induced significant levels of protein damage, already a low cytotoxic concentrations. There was no indication that the toxicity of the tested substances was increased due to metabolic activation in the liver.
This study is classified as acceptable weight of evidence to support the negative genotoxicity profile of the amphoteric, glycinate substances.
- Endpoint:
- in vitro cytogenicity / micronucleus study
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- As part of the weight of evidence there is also data on the related substance Sodium cocopropylenediamine propionate (β-Alanine, N-(2-carboxyethyl)-N-[3-[(2-carboxyethyl)amino]propyl]-, N-C12-18-alkyl derivs., trisodium salts with CAS no 2136366-30-6) which has been tested in the in-vitro micronucleus assay also in human lymphocytes. This substance is closely structurally related to Coco iminodiglycinate, (Amines, N-C12-18 -alkyltrimethylenedi-, reaction products with chloroacetic acid, sodium salts with CAS no 2098351-38-1), the difference being that it contains propyl side chain groups rather than ethyl groups, but is otherwise the same being based on the same diamine.
- Reason / purpose for cross-reference:
- read-across source
- Species / strain:
- lymphocytes: peripheral human lymphocytes
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Conclusions:
- The outcome of an in-vitro micronucleus test, performed according to OECD guideline 487 and GLP principles, showed that Sodium cocopropylenediamine propionate (β-Alanine, N-(2-carboxyethyl)-N-[3-[(2-carboxyethyl)amino]propyl]-, N-C12-18-alkyl derivs., trisodium salts with CAS no 2136366-30-6) is not clastogenic or aneugenic in human lymphocytes under the experimental conditions described in the report.
The in-vitro micronucleus data for Sodium cocopropylenediamine propionate (β-Alanine, N-(2-carboxyethyl)-N-[3-[(2-carboxyethyl)amino]propyl]-, N-C12-18-alkyl derivs., trisodium salts with CAS no 2136366-30-6) is used as supporting data on the genetic toxicity endpoint for the Coco iminodiglycinate, (Amines, N-C12-18-alkyltrimethylenedi-, reaction products with chloroacetic acid, sodium salts with CAS no 2098351-38-1).
This substances are closely structurally related, the difference being that Sodium cocopropylenediamine propionate (β-Alanine, N-(2-carboxyethyl)-N-[3-[(2-carboxyethyl)amino]propyl]-, N-C12-18-alkyl derivs., trisodium salts with CAS no 2136366-30-6) contains propyl side chain groups rather than ethyl groups, but is otherwise the same being based on the same diamine. While it has not been included in the group for other end points there is no reason to believe that this difference in structure could have any influence on possible clastogenic potential.
Referenceopen allclose all
Mutation test 1
strain |
Dose level (µg/plate) |
Metabolic activation |
Mean revertant colony counts |
SD |
TA1535 |
5000 |
- |
16 |
3.2 |
1500 |
- |
10 |
2.6 |
|
500 |
- |
10 |
4.2 |
|
150 |
- |
16 |
2.9 |
|
50 |
- |
17 |
2.5 |
|
0 |
- |
13 |
3.2 |
|
solvent |
- |
18 |
2.1 |
|
5000 |
+ |
18 |
2.5 |
|
1500 |
+ |
15 |
2.6 |
|
500 |
+ |
11 |
6.1 |
|
150 |
+ |
16 |
5.7 |
|
50 |
+ |
13 |
2.5 |
|
0 |
+ |
15 |
1.5 |
|
solvent |
+ |
13 |
4.2 |
|
TA1537 |
5000 |
- |
8 |
3.5 |
1500 |
- |
5 |
2.6 |
|
500 |
- |
8 |
4.7 |
|
150 |
- |
11 |
4.0 |
|
50 |
- |
9 |
2.3 |
|
0 |
- |
10 |
2.6 |
|
solvent |
- |
9 |
2.1 |
|
5000 |
+ |
8 |
1.7 |
|
1500 |
+ |
6 |
3.1 |
|
500 |
+ |
7 |
4.2 |
|
150 |
+ |
10 |
5.9 |
|
50 |
+ |
11 |
1.5 |
|
0 |
+ |
14 |
3.5 |
|
solvent |
+ |
10 |
6.1 |
|
TA1538 |
5000 |
- |
9 |
1.5 |
1500 |
- |
7 |
1.2 |
|
500 |
- |
14 |
2.6 |
|
150 |
- |
12 |
4.0 |
|
50 |
- |
13 |
2.1 |
|
0 |
- |
9 |
6.8 |
|
solvent |
- |
12 |
2.3 |
|
5000 |
+ |
14 |
1.2 |
|
1500 |
+ |
12 |
6.7 |
|
500 |
+ |
11 |
1.7 |
|
150 |
+ |
14 |
3.1 |
|
50 |
+ |
11 |
3.1 |
|
0 |
+ |
19 |
3.2 |
|
solvent |
+ |
20 |
2.1 |
|
TA98 |
5000 |
- |
18 |
3.6 |
1500 |
- |
19 |
3.2 |
|
500 |
- |
20 |
1.5 |
|
150 |
- |
23 |
4.4 |
|
50 |
- |
23 |
3.6 |
|
0 |
- |
28 |
2.6 |
|
solvent |
- |
25 |
5.5 |
|
5000 |
+ |
22 |
1.5 |
|
1500 |
+ |
23 |
2.0 |
|
500 |
+ |
28 |
3.8 |
|
150 |
+ |
27 |
2.6 |
|
50 |
+ |
26 |
2.0 |
|
0 |
+ |
29 |
2.5 |
|
solvent |
+ |
26 |
5.6 |
|
TA100 |
5000 |
- |
121 |
7.4 |
1500 |
- |
122 |
17.3 |
|
500 |
- |
103 |
9.3 |
|
150 |
- |
119 |
12.9 |
|
50 |
- |
113 |
5.3 |
|
0 |
- |
113 |
7.4 |
|
solvent |
- |
108 |
4.6 |
|
5000 |
+ |
112 |
13.6 |
|
1500 |
+ |
118 |
9.8 |
|
500 |
+ |
119 |
17.8 |
|
150 |
+ |
106 |
9.6 |
|
50 |
+ |
108 |
6.1 |
|
0 |
+ |
120 |
4.0 |
|
solvent |
+ |
101 |
7.6 |
- Absence
+ Presence
SD Standard deviation
Positive control Mutation test 1:
Strain |
Compound |
Dose level (µg/plate) |
Metabolic activation |
Mean revertant colony counts |
SD |
TA1535 |
ENNG |
5.0 |
- |
223 |
30.9 |
TA1537 |
9 AC |
80.0 |
- |
X |
- |
TA1538 |
NF |
2.0 |
- |
173 |
28.4 |
TA98 |
NF |
1.0 |
- |
260 |
8.9 |
TA100 |
ENNG |
3.0 |
- |
286 |
6.0 |
TA1535 |
AA |
2.0 |
+ |
139 |
20.6 |
TA1537 |
AA |
2.0 |
+ |
77 |
5.9 |
TA1538 |
NF |
0.5 |
+ |
187 |
15.6 |
TA98 |
AA |
0.5 |
+ |
221 |
19.5 |
TA100 |
AA |
1.0 |
+ |
473 |
72.4 |
- Absence
+ Presence
X Too many colonies to count accurately
SD Standard deviation
ENNG N-ethyl-N’-nitro-N-nitrosoguanidine
9AC 9-aminoacridine
NF 2-nitrofluorene
AA 2-aminoanthracene
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
Justification for classification or non-classification
Based on the general negative in vitro genotoxicity profile for the whole amphoteric, glycinate substance groupSodium cocoamphopolycarboxyglycinate (Amines, N-(3-aminopropyl)-N’-C12-18-alkyltrimethylenedi-, N-(carboxymethyl)derivs., sodium salts with CAS no 2060541-51-5), does not require classification as a mutagen according toRegulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008.
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