Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

No data on acute toxicity is available for"Fatty acids C18 unsat, reaction products with triethylenetetramine" (or TO + TETA). Read-across is performed with "Fatty acids C18 unsat, reaction products with diethylenetetramine" (or TO + DETA).

TO + DETA is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay, is not clastogenic in human lymphocytes, and not mutagenic in the TK mutation test with L5178Y mouse lymphoma cells.

Link to relevant study records

Referenceopen allclose all

Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
experimental study
Adequacy of study:
key study
Study period:
16-jul-2009 to 11-sep-2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.
Qualifier:
according to
Guideline:
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of assay:
in vitro mammalian chromosome aberration test
Species / strain / cell type:
lymphocytes: human peripheral blood
Details on mammalian cell type (if applicable):
- Type and identity of media:
Blood samples
Blood samples were collected by venapuncture using the Venoject multiple sample blood collecting system with a suitable size sterile vessel containing sodium heparin. Immediately after blood collection lymphocyte cultures were started.

Culture medium
Culture medium consisted of RPMI 1640 medium, supplemented with 20% (v/v) heat-inactivated (56°C; 30 min) foetal calf serum, L-glutamine (2 mM), penicillin/streptomycin (50 U/mL and 50 µg/mL respectively) and 30 U/mL heparin.

Lymphocyte cultures
Whole blood (0.4 mL) treated with heparin was added to 5 mL or 4.8 mL culture medium (in the absence and presence of S9-mix, respectively). Per culture 0.1 ml (9 mg/mL) phytohaemagglutinin was added.
Metabolic activation:
with and without
Metabolic activation system:
Rat liver S9-mix induced by a combination of phenobarbital and ß-naphthoflavone
Test concentrations with justification for top dose:
Dose range finding test:
Without S9-mix, 3hr exposure; 24 hr fixation: 10, 20, 30, 50 and 70 µg/ml
Without S9-mix, 24/48hr exposure; 24/48 hr fixation: 1, 3, 10, 20, 30, 100 and 333 µg/ml
With S9-mix, 3hr exposure; 24 hr fixation: 110, 20, 30, 50 and 70 µg/ml
First cytogenetic test:
Without S9-mix, 3 h exposure time, 24 h fixation time:10, 20 and 22 µg/ml
With S9-mix, 3 h exposure, 24 h fixation time: 5, 30 and 35 µg/ml
Second cytogenetic test:
Without S9-mix, 24 hr exposure; 24 hr fixation: 1, 5 and 12 µg/ml
Without S9-mix, 48 hr exposure; 48 hr fixation: 1, 5 and 10 µg/ml
With S9-mix, 3 hr exposure; 48 hr fixation: 10, 35 and 50 µg/ml
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: ethanol
- Justification for choice of solvent/vehicle: Test compound was stable in ethanol and ethanol has been accepted and approved by authorities and international guidelines
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
mitomycin C
Remarks:
without S9 Migrated to IUCLID6: : in Hank's Balanced Salt Solution: 0.5 µg/ml for a 3 h exposure period, 0.2 µg/ml for a 24 h exposure period and 0.1 µg/ml for a 48 h exposure period
Positive control substance:
cyclophosphamide
Remarks:
with S9 Migrated to IUCLID6: : in Hank's Balanced Salt Solution: 10 µg/ml
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium

DURATION
- Preincubation period: 48 hr
- Exposure duration: 3 hr (with and without S9-mix), 24 and 48 hr (without S9-mix)
- Fixation time (start of exposure up to fixation or harvest of cells): 24 and 48 hr

SPINDLE INHIBITOR (cytogenetic assays): colchicine
STAIN (for cytogenetic assays): Giemsa

NUMBER OF REPLICATIONS: duplicates in two independent experiments

NUMBER OF CELLS EVALUATED: 100 metaphase chromosome spreads per culture

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index of each culture was determined by counting the number of metaphases per 1000 cells

OTHER EXAMINATIONS:
- Determination of polyploidy: yes
- Determination of endoreplication: yes
Evaluation criteria:
A test substance was considered positive (clastogenic) in the chromosome aberration test if:
a) It induced a dose-related statistically significant (Chi-square test, one-sided, p < 0.05) increase in the number of cells with chromosome aberrations.
b) A statistically significant and biologically relevant increase in the frequencies of the number of cells with chromosome aberrations was observed in the absence of a clear dose-response relationship.

A test substance was considered negative (not clastogenic) in the chromosome aberration test if none of the tested concentrations induced a statistically significant (Chi-square test, one-sided, p < 0.05) increase in the number of cells with chromosome aberrations.

Statistics:
The incidence of aberrant cells (cells with one or more chromosome aberrations, gaps included or excluded) for each exposure group outside the laboratory historical control data range was compared to that of the solvent control using Chi-square statistics.
Species / strain:
lymphocytes: human peripheral
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: No
- Effects of osmolality: No
- Precipitation: Precipitation in the exposure medium was observed at dose levels of 100 µg/ml and above

RANGE-FINDING/SCREENING STUDIES:
- Toxicity was observed at dose levels of 20 µg/ml and above in the absence and presence of S9, 3 hr treatment/24 hr fixation; at dose levels of 10 µg/ml and above in the absence of S9 for the continuous treatment of 24 hr and at dose levels of 20 µg/ml and above iin the absence of S9 for the continuous treatment of 48 hr

COMPARISON WITH HISTORICAL CONTROL DATA:
- The number of cells with chromosome aberrations found in the solvent and positive control cultures was within the laboratory historical control data range.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
- Appropriate toxicity was reached at the dose levels selected for scoring.

It was noted that Tall oil diethylenetriamine imidazoline increased the number of polyploid cells both in the absence and presence of S9-mix in the first cytogenetic assay and in the presence of S9-mix in the second cytogenetic assay. This may indicate that Tall oil diethylenetriamine imidazoline has the potential to inhibit mitotic processes.

No effects of Tall oil diethylenetriamine imidazoline on the number of cells with endoreduplicated chromosomes were observed both in the absence and presence of S9-mix in both cytogenetic assays.

Conclusions:
Interpretation of results (migrated information):
negative

Tall oil diethylenetriamine imidazoline is not clastogenic in human lymphocytes.
Executive summary:

Evaluation of the ability of Tall oil diethylenetriamine imidazoline to induce chromosome aberrations in cultured peripheral human lymphocytes (with repeat experiment).

This report describes the effect of Tall oil diethylenetriamine imidazoline on the number of chromosome aberrations in cultured peripheral human lymphocytes in the presence and absence of a metabolic activation system (phenobarbital and ß-naphthoflavone induced rat liver S9-mix). The possible clastogenicity of Tall oil diethylenetriamine imidazoline was tested in two independent experiments.

The study procedures described in this report were based on the most recent OECD and EC guidelines.

Batch S000922 of Tall oil diethylenetriamine imidazoline was a clear slightly viscous amber liquid. Tall oil diethylenetriamine imidazoline was dissolved in ethanol.

In the first cytogenetic assay, Tall oil diethylenetriamine imidazoline was tested up to 22 and 35 µg/ml for a 3 h exposure time with a 24 h fixation time in the absence and presence of 1.8% (v/v) S9-fraction, respectively. Appropriate toxicity was reached at these dose levels.

In the second cytogenetic assay, Tall oil diethylenetriamine imidazoline was tested up to 12 µg/ml for a 24 h continuous exposure time with a 24 h fixation time and up to 10 µg/ml for a 48 h continuous exposure time with a 48 h fixation time in the absence of S9-mix. In the presence of S9-mix Tall oil diethylenetriamine imidazoline was tested up to 50 µg/ml for a 3 h exposure time with a 48 h fixation time. Appropriate toxicity was reached at these dose levels.

The number of cells with chromosome aberrations found in the solvent control cultures was within the laboratory historical control data range. Positive control chemicals, mitomycin C and cyclophosphamide, both produced a statistically significant increase in the incidence of cells with chromosome aberrations, indicating that the test conditions were adequate and that the metabolic activation system (S9 -mix) functioned properly.

Tall oil diethylenetriamine imidazoline did not induce a statistically significant or biologically relevant increase in the number of cells with chromosome aberrations in the absence and presence of S9-mix, in either of the two independently repeated experiments.

It was noted that Tall oil diethylenetriamine imidazoline increased the number of polyploid cells both in the absence and presence of S9-mix in the first cytogenetic assay and in the presence of S9-mix in the second cytogenetic assay. This may indicate that Tall oil diethylenetriamine imidazoline has the potential to inhibit mitotic processes.

No effects of Tall oil diethylenetriamine imidazoline on the number of cells with endoreduplicated chromosomes were observed both in the absence and presence of S9-mix in both cytogenetic assays.

Finally, it is concluded that this test is valid and that Tall oil diethylenetriamine imidazoline is not clastogenic in human lymphocytes under the experimental conditions described in this report. Tall oil diethylenetriamine imidazoline may have the potential to disturb mitotic processes and to induce numerical chromosome aberrations.

As explained in the category justification, For cross-reading in general use is made with data of same or lower EA-length where available, and that of Tall oil + DETA representing the worst case.This dossier is for the substance "Fatty acids C18 unsat, reaction products with triethylenetetramine" (or TO + TETA). As for the substance itself no toxicological information is available, cross-reading has been applied to TO + DETA.

Endpoint:
in vitro gene mutation study in mammalian cells
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
18-aug-2009 to 06-oct-2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.
Qualifier:
according to
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)
Deviations:
no
Principles of method if other than guideline:
The recommendations of the “International Workshop on Genotoxicity Tests Workgroup” (the IWGT), published in the literature (Clive et al., 1995, Moore et al., 1999, 2000, 2002, 2003, 2006 and 2007).
GLP compliance:
yes (incl. certificate)
Type of assay:
mammalian cell gene mutation assay
Target gene:
- Thymidine kinase (TK) locus in L5178Y mouse lymphoma cells
Species / strain / cell type:
mouse lymphoma L5178Y cells
Details on mammalian cell type (if applicable):
- Type and identity of media:
-RPMI 1640 Hepes buffered medium (Dutch modification) containing penicillin/streptomycin (50 U/ml and 50 µg/ml, respectively), 1 mM sodium pyruvate and 2 mM L-glutamin supplemented with 10% (v/v) heat-inactivated horse serum (=R10 medium).
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: no
- Periodically "cleansed" against high spontaneous background: yes
Metabolic activation:
with and without
Metabolic activation system:
Rat liver S9-mix induced by a combination of phenobarbital and ß-naphthoflavone
Test concentrations with justification for top dose:
Dose range finding test:
Without and with S9-mix, 3 hours treatment: 1, 3, 10, 33 and 100 µg/mL
Without S9-mix, 24 hours treatment: 0.8, 2.4, 8, 26 and 80 µg/mL
Experiment 1:
Without S9-mix, 3 hours treatment: 0.01, 0.03, 0.1, 0.3, 1, 3, 5 and 6.5 µg/mL
With S9-mix, 3 hours treatment: 1, 4, 8, 10, 15, 30, 40 and 60 µg/mL
Experiment 2
Without S9-mix, 24 hours treatment: 0.24, 0.8, 2.4, 4, 5.2, 6.4, 8 and 8.8 µg/mL
With S9-mix, 3 hours treatment: 0.1, 0.3, 1, 3, 10, 50, 60 and 70 µg/mL
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: ethanol
- Justification for choice of solvent/vehicle: Test compound was stable in ethanol and ethanol has been accepted and approved by authorities and international guidelines
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
methylmethanesulfonate
Remarks:
without S9 Migrated to IUCLID6: 15 µg/mL for the 3 hours treatment period and 5 µg/mL for the 24 hours treatment period
Positive control substance:
cyclophosphamide
Remarks:
with S9 Migrated to IUCLID6: 7.5 µg/ml
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium

DURATION
- Exposure duration:
Short-term treatment
With and without S9-mix: 3 hours
Prolonged treatment period
Without S9-mix: 24 hours
- Expression time (cells in growth medium): 2 days
- Selection time (if incubation with a selection agent): 11 to 12 days

SELECTION AGENT (mutation assays): 5 µg/mL trifluorothymidine (TFT)

NUMBER OF REPLICATIONS:
- Solvent controls: Duplo cultures
- Treatment groups and positive control: Single cultures

NUMBER OF CELLS EVALUATED: 9.6 x 10E5 cells/concentration

DETERMINATION OF CYTOTOXICITY
- Method: relative suspension growth (dose range finding test) and relative total growth (mutation experiments)

RANGE-FINDING/SCREENING STUDIES:
-The suspension growth expressed as the reduction in cell growth after approximately 24 and 48 hours or only 24 hours cell growth, compared to the cell growth of the solvent control, was used to determine an appropriate dose range for the mutagenicity tests

Evaluation criteria:
The global evaluation factor (GEF) has been defined by the IWTG as the mean of the negative/solvent MF distribution plus one standard deviation. For the micro well version of the assay the GEF is 126.

A test substance is considered positive (mutagenic) in the mutation assay if:
a) It induces a MF of more then MF(controls) + 126 in a dose-dependent manner; or
b) In case a repeat experiment is performed when a positive response is observed in one of the tester strains, the positive response should be reproducible in at least one independently repeated experiment.
An observed increase should be biologically relevant and will be compared with the historical control data range.

A test substance is considered equivocal (questionable) in the mutation assay if no clear conclusion for positive or negative result can be made after an additional confirmation study.

A test substance is considered negative (not mutagenic) in the mutation assay if:
a) None of the tested concentrations reaches a mutation frequency of MF(controls) + 126.
b) The results are confirmed in an independently repeated test.

Species / strain:
mouse lymphoma L5178Y cells
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: No
- Effects of osmolality: No
- Precipitation: Precipitation in the exposure medium was observed at dose levels of 26 µg/mL and above

RANGE-FINDING/SCREENING STUDIES:
- Toxicity was observed at dose levels of 10 µg/mL in the absence of S9, 3 hours treatment; at dose levels of 100 µg/mL in the presence of S9, 3 hours treatment; at dose levels of 8 µg/mL in the absence of S9, 24 hours treatment

COMPARISON WITH HISTORICAL CONTROL DATA:
The spontaneous mutation frequencies in the solvent-treated control cultures were between the minimum and maximum value of the historical control data range and within the acceptability criteria of this assay.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
In the absence of S9-mix, the relative total growth of the highest test substance concentration was reduced by 88 and 94% compared to the total growth of the solvent controls after the 3 and 24 hours treatment period, respectively.

In the presence of S9-mix, the relative total growth of the highest test substance concentration was reduced by 92 and 85% compared to the total growth of the solvent controls after the 3 hours treatment period in the first and second experiment, respectively.

Remarks on result:
other: strain/cell type: L5178Y/TK+/-3.7.2C
Remarks:
Migrated from field 'Test system'.
Conclusions:
Interpretation of results (migrated information):
negative

Tall oil diethylenetriamine imidazoline is not mutagenic in the TK mutation test system.
Executive summary:

Evaluation of the mutagenic activity of Tall oil diethylenetriamine imidazoline in an in vitro mammalian cell gene mutation test with L5178Y mouse lymphoma cells (with independent repeat).

This report describes the effects of Tall oil diethylenetriamine imidazoline on the induction of forward mutations at the thymidine-kinase locus (TK-locus) in L5178Y mouse lymphoma cells. The test was performed in two independent experiments in the absence and presence of S9-mix (rat liver S9-mix induced by a combination of phenobarbital and ß-naphthoflavone).

The study procedures described in this report were based on the most recent OECD and EC guidelines.

Batch S000922 of Tall oil diethylenetriamine imidazoline was a clear slightly viscous amber liquid. Tall oil diethylenetriamine imidazoline was dissolved in ethanol.

In the first experiment, Tall oil diethylenetriamine imidazoline was tested up to concentrations of 6.5 and 60 µg/ml in the absence and presence of 8% (v/v) S9-mix, respectively. The incubation time was 3 hours. Toxicity was observed at these dose levels in the absence and presence of S9-mix. Tall oil diethylenetriamine imidazoline was tested up to cytotoxic levels of 88 and 92% in the absence and presence of S9-mix, respectively.

In the second experiment, Tall oil diethylenetriamine imidazoline was tested up to concentrations of 8.8 and 70 µg/ml, in the absence and presence of 12% (v/v) S9-mix, respectively. The incubation times were 24 hours and 3 hours for incubations in the absence and presence of S9-mix, respectively. Tall oil diethylenetriamine imidazoline was tested up to cytotoxic levels of 94 and 85% in the absence and presence of S9-mix, respectively.

The spontaneous mutation frequencies in the solvent-treated control cultures were between the minimum and maximum value of the historical control data range and within the acceptability criteria of this assay.

Mutation frequencies in cultures treated with positive control chemicals were increased by 14-fold for MMS in the absence of S9-mix, and by 9.2- and 8.5-fold for CP in the presence of S9-mix. It was therefore concluded that the test conditions, both in the absence and presence of S9-mix, were appropriate and that the metabolic activation system (S9-mix) functioned properly.

In the absence of S9-mix, Tall oil diethylenetriamine imidazoline did not induce a significant increase in the mutation frequency in the first experiment. This result was confirmed in an independent repeat experiment with modifications in the duration of treatment time.

In the presence of S9-mix, Tall oil diethylenetriamine imidazoline did not induce a significant increase in the mutation frequency in the first experiment. This result was confirmed in an independent repeat experiment with modifications in the concentration of the S9 for metabolic activation.

It is concluded that Tall oil diethylenetriamine imidazoline is not mutagenic in the mouse lymphoma L5178Y test system under the experimental conditions described in this report.

As explained in the category justification, For cross-reading in general use is made with data of same or lower EA-length where available, and that of Tall oil + DETA representing the worst case.This dossier is for the substance "Fatty acids C18 unsat, reaction products with triethylenetetramine" (or TO + TETA). As for the substance itself no toxicological information is available, cross-reading has been applied to TO + DETA.

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:
22 Feb 2007 - 20 Apr 2007
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study has been performed according to OECD and/or EC guidelines and according to GLP principles
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay
Target gene:
- S. typhimurium: Histidine gene
Species / strain / cell type:
other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Metabolic activation system:
S9-fraction from Moltox ((Molecular Toxicology, INC, Boone, NC 28607, USA); liver of rats treated with Aroclor 1254 (500 mg/kg) i.p.
Test concentrations with justification for top dose:
Without S9-mix:
- 0.488, 0.977, 1.95, 3.91, 7.81 and 15.6 µg/plate, for the TA 102 strain in both experiments,
- 3.91, 7.81, 15.6, 31.3, 62.5 and 125 µg/plate, for the TA 1535, TA 1537, TA 98 and TA 100 strains in both experiments.
With S9 mix:
- 7.81, 15.6, 31.3, 62.5, 125 and 250 µg/plate, for the five strains in the first experiment and for the TA 100 and TA 102 strains in the second experiment,
- 15.6, 31.3, 62.5, 125, 250 and 500 µg/plate, for the TA 1535, TA1537 and TA 98 strains in the second experiment.
No precipitate was observed in the Petri plates when scoring the revertants at any dose-levels.
Vehicle / solvent:
Ethanol
Positive controls:
yes
Positive control substance:
other: Sodium-azide 1 µg/plate (TA100, TA1535); 9-Aminoacridine 50 µg/plate (TA1537); 2-Nitrofluorene 0.5 µg/plate (TA98); Mitomycin C 0.5 µg/plate (TA102).
Remarks:
Without metabolic activation
Positive controls:
yes
Positive control substance:
other: 2-Anthramine: 2 µg/plate (TA1535, TA1537, TA98), 10 µg/plate (TA102), 5 µg/plate of Benzo(a)pyrene (TA100)
Remarks:
With metabolic activation
Details on test system and experimental conditions:
DURATION
- Exposure duration: 48 to 72 hours at 37 °C in the dark

NUMBER OF REPLICATIONS:
- Doses of the test substance were tested in triplicate in each strain. Two independent experiments were conducted on each strain with and without S9 mix.

DETERMINATION OF CYTOTOXICITY
Six dose-levels (one plate/dose-level) were tested in the TA 98, TAl00 and TA 102 strains, with and without 89 mix
Based on decrease in the number of revertant colonies and/or a thinning of the bacterial lawn.

OTHER EXAMINATIONS:
In each experiment, the following controls were included using triplicate plates:
- vehicle controls: each bacterial tester strain treated with the vehicle,
- positive controls: each bacterial tester strain treated with appropriate reference mutagens.
The sterility of the S9 mix was checked before the beginning and at the end of each experiment and was found to be satisfactory.
Evaluation criteria:
Acceptance criteria
This study is considered valid if the following criteria are fully met:
- the number of revertants in the vehicle controls is consistent with the historical data of the testing facility,
- the number of revertants in the positive controls is higher than that of the vehicle controls and is consistent with the historical data of the testing facility.
Evaluation criteria
A reproducible 2-fold increase (for the TA 98, TA 100 and TA 102 strains) or 3-fold increase (for the TA 1535 and TA 1537 strains) in the number of revertants compared with the vehicle controls, in any strain at any dose-level and/or evidence of a dose-relationship was considered as a positive result. Reference to historical data, or other considerations of biological relevance may also be taken into account in the evaluation of the data obtained.
Statistics:
Not indicated.
Species / strain:
other: All Salmonella strains tested: TA98, TA100, TA102, TA1535, TA1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
With S9 = 500µg/plate; without S9 = 100µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
The number of revertants for the vehicle and positive controls was as specified in the acceptance criteria. The study was therefore considered valid.

Since the test substance was toxic in the preliminary test, the choice of the highest dose-level was based on the level of toxicity, according to the criteria specified in the international guidelines.

Experiments without S9 mix:
- 0.488, 0.977, 1.95, 3.91, 7.81 and 15.6 µg/plate, for the TA 102 strain in both experiments,
- 3.91, 7.81, 15.6, 31.3, 62.5 and 125 µg/plate, for the TA 1535, TA 1537, TA 98 and TA 100 strains in both experiments.

No precipitate was observed in the Petri plates when scoring the revertants at any dose-levels.
A moderate to marked toxicity was noted at 15.6 µg/plate in the TA 102 strain, at dose-levels = 62.5 µg/plate in the TA 100 strain and at 125 µg/plate in the TA 1535, TA 1537 and TA 98 strains.
The test item did not induce any noteworthy increase in the number of revertants, in any of the five strains.

Experiments with S9 mix:
- 7.81, 15.6, 31.3, 62.5, 125 and 250 µg/plate, for the five strains in the first experiment and for the TA 100 and TA 102 strains in the second experiment,
- 15.6, 31.3, 62.5, 125, 250 and 500 µg/plate, for the TA 1535, TA1537 and TA 98 strains in the second experiment.

No precipitate was observed in the Petri plates when scoring the revertants at any dose-levels.
A moderate to marked toxicity was noted at 250 µg/plate in the TA 98, TA 100 and TA 102 strains and at 500 µg/plate in the TA 1535 strain.
The test item did not induce any noteworthy increase in the number of revertants, in any of the five strains.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Conclusions:
Interpretation of results (migrated information):
negative

Under the experimental conditions of this study, the test substance does not show mutagenic activity in the bacterial reverse mutation test with Salmonella typhimurium.
Executive summary:

The objective of this study was to evaluate the potential of the test item IMIDAZOLINE 4900 (batch No. pilote du 27/07/06, purity: 100%) to induce reverse mutation in Salmonella typhimurium.

The study was performed according to the international guidelines (OECD 471 and Commission Directive No. B13/14) and in compliance with the Principles of Good Laboratory Practice Regulations.

 

Methods

A preliminary toxicity test was performed to define the dose-levels of IMIDAZOLINE 4900 to be used for the mutagenicity study. The test item was then tested in two independent experiments, with and without a metabolic activation system, the S9 mix, prepared from a liver post-mitochondrial fraction (S9 fraction) of rats induced with Aroclor 1254.

Both experiments were performed according to the direct plate incorporation method except for the second test with S9 mix, which was performed according to the preincubation method (60 minutes, 37°C).

Five strains of bacteria Salmonella typhimurium: TA 1535, TA 1537, TA 98, TA 100 and TA 102 were used. Each strain was exposed to at least five dose-levels of the test item (three plates/dose-level). After 48 to 72 hours of incubation at 37°C, the revertant colonies were scored.

The evaluation of the toxicity was performed on the basis of the observation of the decrease in the number of revertant colonies and/or a thinning of the bacterial lawn.

The test item IMIDAZOLINE 4900 was dissolved in ethanol.

 

The dose-levels of the positive controls were as follows:

 

without S9 mix:

- 1 µg/plate of sodium azide (NaN3): TA 1535 and TA 100 strains,

- 50 µg/plate of 9-Aminoacridine (9AA): TA 1537 strain,

- 0.5 µg/plate of 2-Nitrofluorene (2NF): TA 98 strain,

- 0.5 µg/plate of Mitomycin C (MMC): TA 102 strain.

with S9 mix:

- 2 µg/plate of 2-Anthramine (2AM): TA 1535, TA 1537 and TA 98 strains,

- 5 µg/plate of Benzo(a)pyrene (BaP):TA 100 strain,

- 10 µg/plate of 2-Anthramine (2AM): TA 102 strain.

 

Results

The number of revertants for the vehicle and positive controls was as specified in the acceptance criteria. The study was therefore considered valid.

Since the test item was toxic in the preliminary test, the choice of the highest dose-level was based on the level of toxicity, according to the criteria specified in the international guidelines.

 

Experiments without S9 mix:

- 0.488, 0.977, 1.95, 3.91, 7.81 and 15.6 µg/plate, for the TA 102 strain in both experiments,

- 3.91, 7.81, 15.6, 31.3, 62.5 and 125 µg/plate, for the TA 1535, TA 1537, TA 98 and TA 100 strains in both experiments.

No precipitate was observed in the Petri plates when scoring the revertants at any dose-levels.

A moderate to marked toxicity was noted at 15.6 µg/plate in the TA 102 strain, at dose-levels = 62.5 µg/plate in the TA 100 strain and at 125 µg/plate in the TA 1535, TA 1537 and TA 98 strains.

The test item did not induce any noteworthy increase in the number of revertants, in any of the five strains.

 

Experiments with S9 mix:

- 7.81, 15.6, 31.3, 62.5, 125 and 250 µg/plate, for the five strains in the first experiment and for the TA 100 and TA 102 strains in the second experiment,

- 15.6, 31.3, 62.5, 125, 250 and 500 µg/plate, for the TA 1535, TA1537 and TA 98 strains in the second experiment.

No precipitate was observed in the Petri plates when scoring the revertants at any dose-levels.

A moderate to marked toxicity was noted at 250 µg/plate in the TA 98, TA 100 and TA 102 strains and at 500 µg/plate in the TA 1535 strain.

The test item did not induce any noteworthy increase in the number of revertants, in any of the five strains.

 

Conclusion

Under our experimental conditions, the test item IMIDAZOLINE 4900 (batch No. pilote du 27/07/06, purity: 100%) did not show any mutagenic activity in the bacterial reverse mutation test withSalmonella typhimurium. As explained in the category justification, For cross-reading in general use is made with data of same or lower EA-length where available, and that of Tall oil + DETA representing the worst case.

This dossier is for the substance "Fatty acids C18 unsat, reaction products with triethylenetetramine" (or TO + TETA). As for the substance itself no toxicological information is available, cross-reading has been applied to TO + DETA.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

As explained in the category justification, For cross-reading in general use is made with data of same or lower EA-length where available, and that of Tall oil + DETA representing the worst case. This dossier is for the substance "Fatty acids C18 unsat, reaction products with triethylenetetramine" (or TO + TETA). As for the substance itself no toxicological information is available, cross-reading has been applied to TO + DETA.

Tall oil diethylenetriamine imidazoline was tested in theSalmonella typhimuriumreverse mutation assay with five histidine-requiring strains ofSalmonella typhimurium(TA1535, TA1537, TA98, TA100 and TA102). The test was performed in two independent experiments in the presence and absence of S9-mix (rat liver S9-mix induced with Aroclor 1254). The study followed the most recent OECD and EU protocols and was performed under GLP.

There was no significant or dose-related increase in the number of revertant colonies in any of the applied strains, both with and without S9-mix. This was confirmed in an independently repeated experiment.

It is concluded that Tall oil diethylenetriamine imidazoline is not mutagenic in theSalmonella typhimuriumreverse mutation assay.

 

Tall oil diethylenetriamine imidazolinewas studied for its effect on the number of chromosome aberrations in cultured peripheral human lymphocytes in the presence and absence of a metabolic activation system (phenobarbital and ß-naphthoflavone induced rat liver S9-mix), in two independent experiments.

The study was performed under GLP and according to the most recent OECD and EU guidelines.

Tall oil diethylenetriamine imidazolinedid not induce a statistically significant or biologically relevant increase in the number of cells with chromosome aberrations in the absence and presence of S9-mix, in either of the two independently repeated experiments.

It was noted that Tall oil diethylenetriamine imidazoline increased the number of polyploid cells both in the absence and presence of S9-mix in the first cytogenetic assay and in the presence of S9-mix in the second cytogenetic assay. This may indicate that Tall oil diethylenetriamine imidazoline has the potential to inhibit mitotic processes.

No effects of Tall oil diethylenetriamine imidazoline on the number of cells with endoreduplicated chromosomes were observed both in the absence and presence of S9-mix in both cytogenetic assays.

Therefore, it is concluded thatTall oil diethylenetriamine imidazolineis not clastogenic in human lymphocytes.

 

Tall oil diethylenetriamine imidazoline was evaluated for its possible induction of forward mutations at the thymidine-kinase locus (TK-locus) in L5178Y mouse lymphoma cells. The test was performed in two independent experiments in the absence and presence of S9-mix. The study was performed under GLP and according to the most recent OECD and EU guidelines.

In both the presence and absence of S9-mix, Tall oil diethylenetriamine imidazoline did not induce a significant increase in the mutation frequency in the first experiments. This result was confirmed in a repeat experiment with modifications in the duration of treatment time (without S9-mix) or S9 concentration (with S9-mix). Therefore, Tall oil diethylenetriamine imidazoline is not mutagenic in the TK mutation test.

Also other AAI (TEPA, and PolyEA based AAI, including a substance consisting of only Amidoamine without imidazoline) have similarly been tested, with the same results. AAI substances in general therefore are considered to be not genotoxic.

Justification for classification or non-classification

Tall oil diethylenetriamine is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay, is not clastogenic in human lymphocytes, and not mutagenic in the TK mutation test with L5178Y mouse lymphoma cells.

It can therefore be concluded thatTall oil diethylenetriamineis not genotoxic.