(Z)-N-9-octadecenylpropane-1,3-diamine

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

N-Oleyl-1,3-diaminopropane, CAS 7173-62-8, was tested in three in-vitro genotoxicty studies (bacterial mutagenicity, mammalian mutagenicity and mammalian clastogenicity) to current protocol and carried out to GLP with well-defined test substance. All three tests were negative.

Link to relevant study records

Reference

Reference 1

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:

2008-05-14 - 2008-09-23

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Target gene:

histidine operon (his)

Species / strain / cell type:

other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 uvrA

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

liver rat S9-mix (induced with ß-naphthoflavone and phenobarbital)

Test concentrations with justification for top dose:

Experiment 1:
0.00010, 0.000316, 0.00100, 0.00316, 0.0100, 0.0316 and 0.1 µL/plate
Experiment 2:
0.000158, 0.00050, 0.00158, 0.0050, 0.0158, 0.05 and 0.1 µL/plate (TA 98, TA 100, TA 1537, E. coli WP2 uvrA)
0.000050, 0.000158, 0.00050, 0.00158, 0.0050, 0.0158 and 0.05 µL/plate (TA 1535)

Vehicle / solvent:

Ethanol

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 10 µg/plate sodium azide (NaN3) (TA 100, TA 1535); 10 µg/plate (TA 98) and 40 µg/plate (TA 1537) 4-nitro-o-phenylene-diamine (4-NOPD); 1 µL/plate methylmethanesulfonate (MMS) (E. coli); 2.5 µg/plate for TA 98, 100, 1535, 1537 and 10 µg/plate for E. coli

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 hours

NUMBER OF REPLICATIONS: triplicates in 2 experiments

DETERMINATION OF CYTOTOXICITY
- Method: background lawn or reduction in the number of revertants down to a mutation factor of approximately ≤ 0.5 in relation to the solvent control

ADDITIONAL INFORMATION
- Data recording:
The colonies were counted using a ProtoCOL counter (Meintrup DWS Laborgeräte GmbH. If precipitation of the test item precluded automatic counting the revertant colonies were counted by hand. In addition, tester strains with low spontaneous mutation frequency like TA 1535 and TA 1537 were counted manually.

Evaluation criteria:

Criteria of validity:
A test is considered acceptable if for each strain:
1. the bacteria demonstrate their typical response to ampicillin (TA 98, TA 100)
2. the control plates with and without metabolic activation are within the following ranges (mean values of the spontaneous reversion frequency are within the historical control data range):
TA 98: 18 - 54 (-S9) and 18 - 71 (+S9)
TA 100: 75 - 167 (-S9) and 81 - 168 (+S9)
TA 1535: 5 - 29 (-S9) and 6 - 31 (+S9)
TA 1537: 5 - 30 (-S9) and 6 - 36 (+S9)
E. coli WP2 uvrA: 35 - 92 (-S9) and 37 - 101 (+S9)
3. corresponding background growth on both negative control and test plates is observed
4. the positive controls show a distinct enhancement of revertant rates over the control plate

Evaluation of mutagenicity:
The mutation factor is calculated by dividing the mean value of the revertant counts through the mean values of the solvent control (the exact and not the rounded values are used for calculation).
A test item is considered as mutagenic if:
1. a clear and dose-related increase in the number of revertants occurs and/or
2. a biologically relevant positive response for at least one of the dose groups occurs
in at least one tester strain with or without metabolic activation.
A biologically relevant increase is described as follows:
1. if in tester strains Ta 100 and E. coli WP2 uvrA the number of reversions is at least twice as high
2. if in tester strains TA 1535, TA 1537 and TA 98 the number of reversions is at least three times higher
as compared to the reversion rate of the solvent control.

Statistics:

no data

Species / strain:

other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 uvrA

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 0.00316 - 0.1 µL/plate (details in "Additional information on results")

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: No precipitation of the test item was observed in any of the 5 tester strains used in experiment 1 and 2 with and without metabolic activation.

RANGE-FINDING/SCREENING STUDIES:
The toxicity of the test item was determined with tester strain TA 98 and TA 100 in a pre-experiment. Eight concentrations were tested for toxicity and induction of mutations with three plates each. The experimental conditions in this pre-experiment were the same as for the main experiment.
Toxicity may be detected by a clearing or rather diminution of the background lawn or a reduetion in the number of revertants down to a mutation factor of approximately ≤ 0.5 in relation to the solvent control.
The test item was tested in the pre-experiment at the following concentrations:
0.00316, 0.0100, 0.0316, 0.100, 0.316, 1.0, 2.5 and 5.0 µL/plate
Based on the pre-experiment results the following concentrations were used for the main experiments:
Experiment 1: 0.0001 - 0.1 µL/plate and Experiment 2: 0.000158 - 0.1 µL/plate (TA 98, TA 100, TA 1537, E. coli WP2 uvrA) and 0.00005 - 0.05 µL/plate (TA 1535)

ADDITIONAL INFORMATION ON CYTOTOXICITY:
Cytotoxic effects in experiment 1:
at 0.00316 µL/plate (-S9) and 0.0316 µL/plate (+S9) (TA 98 and TA 100);
at 0.010 µL/plate (-S9) and 0.1 µL/plate (+S9) (TA 1535);
at 0.01 µL/plate (-S9) and 0.0316 µL/plate (+S9) (TA 1537);
at 0.0316 µL/plate (-S9) and 0.1 µL/plate (+S9) (E. coli WP2 uvrA)

Cytotoxic effects in experiment 2:
at 0.0050 µL/plate (-S9) and 0.05 µL/plate (+S9) (TA 98, TA 100, TA 1535, TA 1537);
at 0.0158 µL/plate (-S9) and 0.1 µL/plate (+S9) (E. coli WP2 uvrA)

Remarks on result:

other: strain/cell type: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 uvrA

Remarks:

Migrated from field 'Test system'.

Table 1: Ames Test Results - Experiment 1

With or without S9-Mix	Test substance concentration (dose/plate)	Mean number of revertant colonies per plate (triplicates)
		Base-pair substitution type			Frameshift type
		TA 100	TA 1535	E. coli WP2 uvrA	TA 98	TA 1537
-	A. dest.	95	20	53	28	6
-	Vehicle control (EtOH)	102	23	46	24	6
-	0.000100µL	96	17	42	29	9
-	0.000316 µL	67	16	45	23	10
-	0.00100 µL	94	26	46	26	11
-	0.00316 µL	67	24	46	24	7
-	0.0100 µL	41	13	53	15	4
-	0.0316 µL	0	0	0	0	0
-	0.1 µL	0	0	0	0	0
Positive controls - S9	Name	NaN3	NaN3	MMS	4-NOPD	4-NOPD
	Concentrations (μg/plate)	10 µg	10 µg	1 µL	10 µg	40 µg
	Number of colonies/plate	1257	1145	407	395	99
		TA 100	TA 1535	E. coli WP2 uvrA	TA 98	TA 1537
+	A. dest.	99	19	56	33	10
+	Vehicle control (EtOH)	97	23	57	38	11
+	0.000100µL	108	29	56	40	9
+	0.000316 µL	103	23	46	36	7
+	0.00100 µL	112	25	63	44	8
+	0.00316 µL	114	26	62	43	11
+	0.0100 µL	129	25	69	41	13
+	0.0316 µL	62	13	66	21	6
+	0.1 µL	0	0	15	0	0
Positive controls + S9	Name	2-AA	2-AA	2-AA	2-AA	2-AA
	Concentrations (μg/plate)	2.5 µg	2.5 µg	10 µg	2.5 µg	2.5 µg
	Number of colonies/plate	2209	243	204	3114	211

Table 2: Ames Test Results - Experiment 2

With or without S9-Mix	Test substance concentration (dose/plate)	Mean number of revertant colonies per plate (triplicates)
		Base-pair substitution type			Frameshift type
		TA 100	TA 1535	E. coli WP2 uvrA	TA 98	TA 1537
-	A. dest.	134	26	54	21	7
-	Vehicle control (EtOH)	134	31	51	30	7
-	0.000050µL	-	32	-	-	-
-	0.000158µL	126	24	56	29	9
-	0.0005 µL	124	28	53	23	7
-	0.00158 µL	117	19	50	22	12
-	0.005 µL	75	15	50	26	8
-	0.0158 µL	13	0	35	3	0
-	0.05 µL	0	0	0	0	0
-	0.1 µL	0	-	0	0	0
Positive controls - S9	Name	NaN3	NaN3	MMS	4-NOPD	4-NOPD
	Concentrations (μg/plate)	10 µg	10 µg	1 µL	10 µg	40 µg
	Number of colonies/plate	1519	1537	447	528	112
		TA 100	TA 1535	E. coli WP2 uvrA	TA 98	TA 1537
+	A. dest.	105	21	56	39	13
+	Vehicle control (EtOH)	81	22	53	37	9
+	0.000050µL	-	28	-	-	-
+	0.000158µL	108	29	67	37	11
+	0.0005 µL	89	25	64	42	6
+	0.00158 µL	97	25	61	45	8
+	0.005 µL	103	17	51	40	8
+	0.0158 µL	112	22	50	41	17
+	0.05 µL	24	0	66	0	0
+	0.1 µL	0	-	2	0	0
Positive controls + S9	Name	2-AA	2-AA	2-AA	2-AA	2-AA
	Concentrations (μg/plate)	2.5 µg	2.5 µg	10 µg	2.5 µg	2.5 µg
	Number of colonies/plate	1611	181	231	2440	328

NaN3= Sodium azide

MMS = Methyl methane sulfonate

4-NOPD = 4-Nitro-o-phenylene-diamine

2-AA = 2-Aminoanthracene

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, N-Oleyl-1,3- diaminopropane did not cause gene mutations by base pair changes or frameshifts in the genome of the tester strains used.
Therefore, N-Oleyl-1,3-diaminopropane is considered to be non-mutagenic in this bacterial reverse mutation assay.

Executive summary:

The test item N-Oleyl-1,3-diaminopropane was investigated for its potential to induce gene mutations according to the OECD guideline 471.

In two independent plate incorporation tests using Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and tester strain E. coli WP2 uvrA several concentrations of the test item were used. Each assay was conducted with and without metabolic activation. The concentrations, including the controls, were tested in triplicates. The following concentrations of the test item were prepared and used in the experiments:

Experiment 1:

0.00010, 0.000316, 0.00100, 0.00316, 0.0100, 0.0316 and 0.1 µL/plate

Experiment 2:

0.000158, 0.00050, 0.00158, 0.0050, 0.0158, 0.05 and 0.1 µL/plate (TA 98, TA 100, TA 1537, E. coli WP2 uvrA) and

0.000050, 0.000158, 0.00050, 0.00158, 0.0050, 0.0158 and 0.05 µL/plate (TA 1535).

No precipitation of the test item was observed in any of the five tester strains used in experiment 1 and 2 with and without metabolic activation.

Toxic effects of the test item were noted in all tester strains evaluated in experiment 1 and 2.

In experiment 1 toxic effects of the test item were observed in tester strains TA 98 and TA 100 at doses of 0.00316 µL/plate and higher (without metabolic activation) and at doses of 0.0316 µL/plate and higher (with metabolic activation). In tester strain TA 1535 toxic effects of the test item were noted at doses of 0.01 µL/plate and higher (without metabolic activation) and at a dose of 0.1 µL/plate (with metabolic activation). In tester strain TA 1537 toxic effects of the test item were seen at doses of 0.01 µL/plate and higher (without metabolic activation) and at doses of 0.0316 µL/plate and higher (with metabolic activation). In tester strain E. coli WP2 uvrA toxic effects of the test item were noted at doses of 0.0316 µL/plate and higher (without metabolic activation) and at a dose of 0.1 µL/plate (with metabolic activation).

In experiment 2 toxic effects of the test item were noted in tester strains TA 98, TA 100 and TA 1537 at doses of 0.005 µL/plate and higher (without metabolic activation) and at doses of 0.05 µL/plate and higher (with metabolic activation). In tester strain TA 1535 toxic effects of the test item were observed at doses of 0.005 µL/plate and higher (without metabolic activation) and at a dose of 0.05 µL/plate (with metabolic activation). In tester strain E. coli WP2 uvrA toxic effects of the test item were seen at doses of 0.0158 µL/plate and higher (without metabolic activation) and at a dose of 0.1 µL/plate (with metabolic activation).

No biologically relevant increases in revertant colony numbers of any of the five tester strains were observed following treatment with N-Oleyl-1,3-diaminopropane at any concentration level, neither in the presence nor absence of metabolic activation in experiment 1 and 2.

The reference mutagens induced a distinct increase of revertant colonies indicating the validity of the experiments.

Reference 2

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:

2008-05-14 - 2008-10-08

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

in vitro mammalian chromosome aberration test

Target gene:

not applicable

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

Details on mammalian cell type (if applicable):

- Type and identity of media: MEM (minimum essential medium) supplemented with 10% FCS (foetal calf serum)

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

rat liver S9-mix (induced with β-naphthoflavone and phenobarbital)

Test concentrations with justification for top dose:

Experiment I:
+S9: 0.5, 1.0, 2.0, 4.0, 5.0, 6.5 and 8.0 µg/mL
-S9: 0.2, 0.4, 0.55, 0.7, 0.85, 1.0 and 1.2 µg/mL

Experiment II:
+S9: 1.5, 2.0, 2.5, 3.0, 3.5, 4.0 and 4.5 µg/mL
-S9: 0.05, 0.1, 0.2, 0.4, 0.55, 0.7, 0.85, 1.0 and 1.2 µg/mL

Vehicle / solvent:

Ethanol

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 400 and 900 µg/mL ethylmethanesulphonate (EMS) and 0.83 µg/mL cyclophosphamide (CPA)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Exposure duration: 4 and 20 hours
- Fixation time (start of exposure up to fixation or harvest of cells): 20 hours

SPINDLE INHIBITOR (cytogenetic assays): Colcemid (0.2 µg/mL)
STAIN (for cytogenetic assays): Giemsa

NUMBER OF REPLICATIONS: duplicates in 2 independent experiments

NUMBER OF CELLS EVALUATED: 200 per concentration

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index

OTHER EXAMINATIONS:
- Determination of polyploidy: yes

Evaluation criteria:

The chromosomal aberration assay is considered acceptable if it meets the following criteria:
- the number of aberration found in the negative and/or solvent controls falls within the range of historical laboratory control data: 0.0% - 4.5% (+S9) resp. 0.0% - 4.0% (-S9)
- the positive control substances should produce biologically relevant increases in the number of cells with structural chromosome aberrations

Criteria for determinig a positive result:
- a clear and dose-related increase in the number of cells with aberrations,
- a biologically relevant response for at least one of the dose groups, which is higher than the laboratory negative control range (up to 4.5% aberrant cells (+S9) resp. 4.0% aberrant cells (-S9))

Statistics:

no data

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Experiment I: at 1.0 µg/mL and higher (-S9) and 4.0 µg/mL and higher (+S9); Experiment II: at 0.4 µg/mL and higher (-S9) and 4.0 µg/mL and higher (+S9)

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES:
The concentration range for the Preliminary Toxicity Test was 0.0078 to 5000 µg/mL without metabolic activation and 1.95 to 5000 µg/mL with metabolic activation. The test item could be dissolved at a concentration of 500 µg/mL in ethanol. After dilution with cell culture medium, precipitation of the test item appeared in a concentration of 15.6 µg/mL and higher.
The selection of the concentrations used in experiment I and II based on data from the solubility and the pre-experiment which were performed according to guidelines. In experiment I (-S9) 1.2 µg/mL and (+S9) 4.0 µg/mL were selected as highest dose groups for the microscopic analysis of chromosomal aberrations. And in experiment II (-S9) 0.4 µg/mL and (+S9) 4.5 µg/mL were selected as highest dose groups for the microscopic analysis of chromosomal aberrations.

COMPARISON WITH HISTORICAL CONTROL DATA: within historical reference range

Remarks on result:

other: strain/cell type: Chinese hamster lung fibroblasts (V79)

Remarks:

Migrated from field 'Test system'.

Summary of aberration rates in experiment I

Dose Group	Concentration [µg/mL]	Treatment Time	Fixation Interval	Mean % aberrant cells
				incl. Gaps	excl. Gaps
without metabolic activation
C	0	4 h	20 h	2.0	0.0
S	0	4 h	20 h	4.5	1.5
5	0.85	4 h	20 h	3.5	1.0
6	1.0	4 h	20 h	0.5	0.0
7	1.2	4 h	20 h	0.5	0.0
EMS	900	4 h	20 h	11.5	8.5
with metabolic activation
C	0	4 h	20 h	5.0	3.5
S	0	4 h	20 h	2.0	1.5
2	1.0	4 h	20 h	4.5	1.5
3	2.0	4 h	20 h	4.5	2.0
4	4.0	4 h	20 h	3.5	1.0
CPA	0.83	4 h	20 h	10.5	9.0

Summary of aberration rates in experiment II

Dose Group	Concentration [µg/mL]	Treatment Time	Fixation Interval	Mean % aberrant cells
				incl. Gaps	excl. Gaps
without metabolic activation
C	0	20 h	20 h	3.0	1.5
S	0	20 h	20 h	3.5	2.0
5	0.1	20 h	20 h	4.0	2.5
6	0.2	20 h	20 h	2.0	0.5
7	0.4	20 h	20 h	3.0	1.0
EMS	400	20 h	20 h	12.0	8.0
with metabolic activation
C	0	4 h	20 h	4.0	1.5
S	0	4 h	20 h	4.5	2.5
2	2.5	4 h	20 h	3.5	1.5
3	4.0	4 h	20 h	3.0	1.5
4	4.5	4 h	20 h	2.5	1.0
CPA	0.83	4 h	20 h	11.5	8.5

200 cells evaluated for each concentration

C: Negative control (culture medium)

S: Solvent control (Ethanol)

EMS: Positive control (-S9: ethylmethansulfonate)

CPA: Positive control (+S9: cyclophosphamide)

Conclusions:

Interpretation of results (migrated information):
negative

In conclusion, it can be stated that during the described in vitro chromosomal aberration test and under the experimental conditions reported, the test item N-Oleyl-1,3-diaminopropane did not induce structural chromosome aberrations in the V79 Chinese hamster cell line. Therefore, the test item N-Oleyl-1,3-diaminopropane is considered to be non-clastogenic.

Executive summary:

The test item N-Oleyl-1‚3-diaminopropane was investigated for a possible potential to induce structural chromosomal aberrations in V79 cells of the Chinese hamster in vitro in the absence and presence of metabolic activation with S9 homogenate.
The selection of the concentrations used in experiment I and II based on data from the solubility test and the pre-experiment which were performed according to the guidelines.
In experiment I without metabolic activation 1.2 µg/mL and with metabolic activation 4.0 µg/mL were selected as highest dose groups for the microscopic analysis of chromosomal aberrations. In experiment II without metabolic activation 0.4 µg/mL and with metabolic activation 4.5 µg/mL were selected as highest dose groups for the microscopic analysis of chromosomal aberrations.
The chromosomes were prepared 20 h after start of treatment with the test item. The treatment intervals were 4 h with and without metabolic activation (experiment I) and 4 h with and 20 h without metabolic activation (experiment II). Two parallel cultures were set up. 100 metaphases per culture were scored for structural chromosomal aberrations.
The following concentrations were evaluated for microscopic analysis:
Experiment I:
with metabolic activation: 1.0, 2.0 and 4.0 µg/mL
without metabolic activation: 0.85, 1.0 and 1.2 µg/mL
Experiment II:
with metabolic activation: 2.5, 4.0 and 4.5 µg/mL
without metabolic activation: 0.1, 0.2 and 0.4 µg/mL
Precipitation:
No precipitation of the test item was noted with and without metabolic activation after the incubation at the concentrations evaluated.
Toxicity:
In experiment I without metabolic activation, a biologically relevant decrease of the relative mitose index (decrease below 70% rel. mitose index) was noted at 1.0 µg/mL and higher (48% at 1.0 µg/mL, 40% at 1.2 µg/mL). The cell density was not decreased. With metabolic activation a biologically relevant decrease of the relative mitose index (decrease below 70% rel. mitose index) was noted at a concentration of 4.0 µg/mL (47% at 4 µg/mL). The cell density was also decreased at this concentration (61%).
In experiment II without metabolic activation, a biologically relevant decrease of the relative mitose index (decrease below 70% rel. mitose index) was noted at 0.4 µg/mL (49%). The cell density was also decreased (65%). With metabolic activation, a biologically relevant decrease of the relative mitose index (decrease below 70% rel. mitose index) was noted at 4.0 µg/mL and higher (46% at 4.0 µg/mL, 30% at 4.5 g/mL). No decrease of the cell density was noted up to the highest dose evaluated.
Clastogenicity:
In the experiment without metabolic activation the aberration rates of the negative control (0.0%) and the solvent control (1.5%) were within the historical control data of the negative control (0.0% - 4.0%). The aberration rates of all dose groups evaluated were within the range of the historical control data. Mean values of 1.0% (0.85 µg/mL), 0.0% (1 and 1.2 µg/mL) aberrant cells were found.
In the experiment with metabolic activation the aberration rates of the negative control (3.5%) and the solvent control (15%) were within the historical control data (0.0% - 4.5%). The aberration rate of all dose groups evaluated were within the range of the historical control data. Mean values of 1.5% (1 µg/mL), 2.0% (2 µg/mL) and 1.0% (4 µg/mL) aberrant cells were found.
In experiment II without metabolic activation the aberration rate of the negative control (1.5%), the solvent control (2.0%) and all dose groups treated with the test item (2,5% (0.1 µg/mL, 0.5% (0.2 µg/mL) and 1.0% (0.4 µg/mL)) were within the historical control data of the testing facility (0.0% - 4.0%). With metabolic activation the aberration rates of the negative control (1.5%), the solvent control (2.5%) and all dose groups treated with the test item (1.5% (2.5 µg/mL), 1.5% (4.0 µg/mL) and 1.0% (4.5 µg/mL)) were within the historical control data of the testing facility (0.0% - 4.5%). The number of aberrant cells found in the dose groups treated with the test item did not show a biologically relevant increase compared to the corresponding negative control, In addition, no dose-response relationship was observed.
Polyploid cells
No biologically relevant increase in the frequencies of polyploid cells was found after treatment with the test item.
EMS (400 and 900 µg/mL) and CPA (0.83 µg/mL) were used as positive controls and induced distinct and biologically relevant increases in cells with structural chromosomal aberration.

Reference 3

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:

2008-05-14 - 2010-04-29

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

mammalian cell gene mutation assay

Target gene:

Hypoxanthine-guanine-phosphoribosyl-transferase (HPRT)

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

Details on mammalian cell type (if applicable):

- Type and identity of media: MEM (minimal essential medium)
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

rat liver S9-mix (indued with β-naphthoflavone and phenobarbital)

Test concentrations with justification for top dose:

Experiment I:
-S9: 0.350, 0.425, 0.500, 0.575, 0.650, 0.725, 0.800, 0.875 µg/mL
+S9: 0.05, 0.10, 0.25, 0.5, 1.0, 3.0, 4.0, 5.0 µg/mL

Experiment II:
-S9: 0.1, 0.2, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 µg/mL
+S9: 1.0, 2.0, 3.8, 4.2, 5.0, 5.5, 6.0, 7.0 µg/mL

Vehicle / solvent:

Ethanol

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 300 µg/mL ethylmethanesulphonate (EMS); 1.0 µg/mL (Experiment I) and 1.5 µg/mL (Experiment II) 7,12-dimethylbenzanthracene(DMBA)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Exposure duration: 4, 20 hours
- Expression time (cells in growth medium): 48 to 72 hours after treatment

SELECTION AGENT (mutation assays): thioguanine (TG)

NUMBER OF REPLICATIONS: two independent experiments

DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency; relative total growth

Evaluation criteria:

A mutation assay is considered acceptable if it meets the following criteria:
- negative and/or solvent controls fall within the performing laboratories historical control data range: 1 - 39 mutants/10E6 cells
- the absolute cloning efficiency: ([number of positive cultures x 100] / total number of seeded cultures) of the negative and/or solvent controls is > 50%
- the spontaneous mutant frequency in the negative and/or solvent controls is in the range of historical control data
- the positive controls (EMS and DMBA) induce significant increases (at least 3-fold increase of mutant frequencies related to the comparable negative control values and higher than the historical range of negative controls) in the mutant frequencies.

Atest is considered negative if there is no biological relevant increase in the number of mutants. There are several criteria for determining a positive result:
- a reproducible 3-times higher mutation frequency than the solvent control for at least one of the concentrations
- a concentration related increase of the mutation frequency; such an evaluation may be considered also in the case that a 3-fold increase of the mutant frequency is not observed.

Statistics:

No data

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Experiment I: at 0.875 µg/mL (-S9) and at 5.0 µg/mL (+S9); Experiment II: at 0.9 µg/mL (-S9) and at 7.0 µg/mL (+S9)

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES:
The toxicity of the test item was based on data from pre-experiment. Eight concentrations were tested: 0.0625 - 4.0 µg/mL (-S9) and 0.125 - 3.0 µg/mL (+S9).
In experiment I 0.875 µg/mL (-S9) and 5 µg/mL (+S9) were selected as the highest concentrations. In experiment II 0.9 µg/mL (-S9) and 7 µg/mL [+S9) were selected as the highest concentrations. Experiment II without metabolic activation was performed as a 20 h long-term exposure assay.

COMPARISON WITH HISTORICAL CONTROL DATA:
All values of the negative controls and test item concentrations found were within the historical control data

Remarks on result:

other: strain/cell type: Chinese hamster lung fibroblasts (V79)

Remarks:

Migrated from field 'Test system'.

Table 1: Experiment I - without metabolic activation

Dose Group	Concentration [µg/mL]	Relative Growth [%]	Factor* (survived cells / seeded cells)	Mutant colonies per 10E6 cells	Mutation factor
NC1	0 0	125.1	0.78	11.48
NC2		133.3	0.82	14.58
S1	0 0	100.0 100.0	0.78	14.08
S2			0.69	6.48
5	0.350	104.1	0.73	18.49	1.80
6	0.425	98.6	0.71	25.42	2.47
7	0.500	90.9	0.81	8.07	0.79
8	0.575	80.5	0.81	24.66	2.40
9	0.650	53.5	0.75	22.09	2.15
10	0.725	51.9	0.74	14.29	1.39
11	0.800	33.2	0.74	24.19	2.35
12	0.875	13.6	0.63	3.96	0.39
EMS	300	105.9	0.82	150.37	14.62

Table 2: Experiment I - with metabolic activation

Dose Group	Concentration [µg/mL]	Relative Growth [%]	Factor* (survived cells / seeded cells)	Mutant colonies per 10E6 cells	Mutation factor
NC1	0 0	103.0	0.97	10.80
NC2		114.6	0.69	28.34
S1	0 0	100.0 100.0	0.84	11.96
S2			0.87	7.51
1	0.05	97.8	0.83	21.79	2.24
2	0.10	82.0	0.72	13.81	1.42
3	0.25	83.9	0.84	19.69	2.02
4	0.5	79.8	0.83	7.85	0.81
5	1.0	81.3	0.71	21.86	2.25
6	3.0	68.5	0.73	17.08	1.75
7	4.0	47.2	0.72	22.85	2.35
8	5.0	10.1	0.57	14.13	1.45
DMBA	1.0	67.0	0.68	147.19	15.12

Table 3: Experiment II - without metabolic activation

Dose Group	Concentration [µg/mL]	Relative Growth [%]	Factor* (survived cells / seeded cells)	Mutant colonies per 10E6 cells	Mutation factor
NC1	0 0	115.5	0.87	5.74
NC2		105.7	0.90	11.67
S1	0 0	100.0 100.0	0.87	5.76
S2			0.75	10.65
5	0.1	80.6	0.56	8.06	0.98
6	0.2	80.6	0.56	11.71	1.43
7	0.4	67.9	0.76	1060	1.29
8	0.5	62.0	0.51	13.70	1.67
9	0.6	38.2	0.94	8.55	1.04
10	0.7	27.7	0.84	12.54	1.53
11	0.8	26.7	0.67	2.98	0.36
12	0.9	12.6	0.99	9.06	1.10
EMS	300	49.3	0.57	184.12	22.44

Table 4: Experiment II - with metabolic activation

Dose Group	Concentration [µg/mL]	Relative Growth [%]	Factor* (survived cells / seeded cells)	Mutant colonies per 10E6 cells	Mutation factor
NC1	0 0	118.5	0.75	16.02
NC2		96.0	0.68	8.77
S1	0 0	100.0 100.0	0.86	16.36
S2			0.84	1.20
2	1.0	95.0	0.70	9.34	1.06
3	2.0	95.0	0.68	4.39	0.50
5	3.8	85.0	0.72	6.23	0.71
6	4.2	80.0	0.74	6.11	0.70
7	5.0	70.0	0.78	3.19	0.36
8	5.5	65.0	0.55	2.75	0.31
9	6.0	40.0	0.75	8.70	0.99
10	7.0	16.5	0.79	5.06	0.58
DMBA	1.5	80.0	0.87	116.09	13.23

NC: negative control / medium control

SC: solvent control (ethanol)

*: cloning efficiency x cells seeded

EMS: Ethylmethansulfonate

DMBA: 7,12 -Dimethylbenz(a)anthracene

Conclusions:

Interpretation of results (migrated information):
negative

In conclusion, in the described in vitro cell gene mutagenicity test under the experimental conditions reported, the test item N-Oleyl-1,3-diaminopropane is considered to be non-mutagenic in the HPRT locus using V79 cells of the Chinese hamster.

Executive summary:

The test item N-Oleyl-1,3-diaminopropane was assessed for its potential to induce gene mutations at the HPRT locus using V79 cells of the Chinese hamster according to the OECD guideline 476.
The main experiments were carried out without and with metabolic activation. The experiments with metabolic activation were performed by including liver microsomes and NADP for efficient detection of a wide variety of carcinogens requiring metabolic activation.
The selection of the concentrations used in the main experiments was based on data from the pre-experiments according to the OECD guideline 476.
In experiment I 0.875 µg/mL (without metabolic activation) and 5.0 µg/mL (with metabolic activation) were selected as the highest concentrations. In experiment II 0.9 µg/mL (without metabolic activation) and 7.0 µg/mL (with metabolic activation) were selected as the highest concentrations. Experiment II without metabolic activation was performed as a 20 h long-term exposure assay.
The pH-value detected with the test item was within the physiological range. The test item was investigated at the following concentrations:
Experiment I
without metabolic activation:
0.350, 0.425, 0.500, 0.575, 0.650, 0.725, 0.800 and 0.875 µg/mL
and with metabolic activation:
0.05, 0.10, 0.25,0.5,1.0, 3.0, 4.0 and 5.0 µg/mL
Experiment II
without metabolic activation:
0.1, 0.2, 0.4, 0.5, 0.6, 0.7, 0.8 and 0.9 µg/mL
and with metabolic activation:
1.0, 2.0, 3.8, 4.2, 5.0, 5.5, 6.0 and 7.0 µg/mL
No precipitation oft he test item was noted in experiment I and experiment II.
Toxicity:
A biologically relevant growth inhibition (reduction of relative growth below 70%) was observed after the treatment with the test item in experiment I and II with and without metabolic activation.
In experiment I without metabolic activation the relative growth was 13.6% for the highest concentration (0.875 µg/mL) evaluated. The highest biologically relevant concentration evaluated with metabolic activation was 5.0 µg/mL with a relative growth of 10.1%.
In experiment II without metabolic activation the relative growth was 12.6% for the highest concentration (0.9 µg/mL) evaluated. The highest biologically relevant concentration evaluated with metabolic activation was 7.0 µg/mL with a relative growth of 16 .5%.

Mutagenicity:
In experiment I without metabolic activation mutant values of the negative controls and test item concentrations found were within the historical control data of the test facility BSL BIOSERVICE (about 1 - 39 mutants per 10⁶ cells). No dose-response relationship could be observed. The mutation frequencies found in the groups treated with the test item did not show a biologically relevant increase as compared to the negative controls.
Mutation frequencies with the negative control were found to be 11.48 and 14.58 mutants/10⁶ cells, 14.08 and 6.48 mutants/10⁶ cells for the solvent control and in the range of 3.96 to 25.42 mutants/10⁶ cells with the test item, respectively. The highest mutation rate (compared to the solvent control values) of 2.47 was found at a concentration of 0.425 µg/mL with a relative growth of 98.6%.
With metabolic activation all mutant values of the negative controls and test item concentrations found were within the historical control data of the test facility BSL BIOSERVICE (about 2 - 28 mutants per 10⁶ cells). No dose-response relationship could be observed. The mutation frequencies found in the groups treated with the test item did not show a biologically relevant increase as compared to the solvent controls.
Mutation frequencies of the negative control were found to be 10.80 and 28.34 mutants/10⁶ cells, 11.96 and 7.51 mutants/10⁶ cells for the solvent control and in the range of 7.85 to 22.85 mutants/10⁶ cells with the test item, respectively. The highest mutation rate (compared to the solvent control values) of 2.35 was found at a concentration of 4.0 µg/mL with a relative growth of 47.2%.
In experiment II without metabolic activation all mutant values found were within the historical control data of the test facility BSL BIOSERVICE (about 1 - 39 mutants per 10⁶ cells). No dose-response relationship could be observed. The mutation frequencies found in the groups treated with the test item did not show a biologically relevant increase as compared to the solvent controls.
Mutation frequencies with the negative control were found to be 5.47 and 11.67 mutants/10⁶ cells, 5.76 and 10.65 mutants/10⁶

cells for the solvent control and in the range of 2.98 to 13.70 mutants/10⁶ cells with the test item, respectively. The highest mutation rate (compared to the solvent controls values) of 1.67 was found at a concentration of 0.5 µg/mL with a relative growth of 62.0%.
In experiment II with metabolic activation most mutant values found were within the historical control data of the test facility BSL BIOSERVICE (about 2 - 28 mutants per 10⁶ cells). No dose-response relationship could be observed. The mutation frequencies found in the groups treated with the test item did not show a biologically relevant increase as compared to the solvent controls.
Mutation frequencies of the negative control were found to be 16.02 and 8.77 mutants/10⁶ cells, 16.36 and 1.20 mutants/10⁶ cells for the solvent control and in the range of 2.75 to 9.34 mutants/10⁶ cells with the test item, respectively. The highest mutation rate (compared to the solvent control values) of 1.06 was found at a concentration of 1.0 µg/mL with a relative growth of 95%.
DMBA (1.0 and 1.5 µg/mL) and (300 µg/mL) were used as positive controls and showed distinct and biologically relevant effects in mutation frequency.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Mode of Action Analysis / Human Relevance Framework

Based on structure and mechanism of cytotoxicity, genototoxicity by alkyl-diamines is not expected. In physiological circumstances, the diamines have a cationic surfactant structure which leads to high adsorptive properties to negatively charged surfaces as cellular membranes. The apolar tails easily dissolve in the membranes, whereas the polar head causes disruption and leakage of the membranes leading to cell damage or lysis of the cell content. As a consequence, the whole molecule will not easily pass membrane structures. Noteworthy in this respect is that recent research shows that the log distribution coefficient for cationic surfactants between water and phospholipid are possibly several orders of magnitude higher than between water and oil.

 

Cytotoxicity through disruption of cell membrane will occur rather than absorption over the cell membrane into the cell and transfer to the nucleus to interact with DNA.

Additional information

N-Oleyl-1,3-diaminopropane, CAS 7173-62-8,was tested for genetic toxicity in three in vitro tests. In a bacterial reverse mutation assay the substance was not mutagenic with or without S9 mix. It was also tested for mammalian cell gene mutations at the HPRT locus using V79 cells of the Chinese hamster and was not mutagenic. In addition, the substance was tested in a chromosomal aberration test in human lymphocytes and was found to be not clastogenic. All tests were to current OECD/EU protocols carried out to GLP and with a clearly defined and described test substance. Based on these results it can be concluded that N-Oleyl-1,3-diaminopropane is not to be expected to be a genotoxic hazard to human health.

 

 

Based on structure and mechanism of cytotoxicity, genototoxicity by alkyl-diamines is not expected. In physiological circumstances, the diamines have a cationic surfactant structure which leads to high adsorptive properties to negatively charged surfaces as cellular membranes. The apolar tails easily dissolve in the membranes, whereas the polar head causes disruption and leakage of the membranes leading to cell damage or lysis of the cell content. As a consequence, the whole molecule will not easily pass membrane structures. Cytotoxicity through disruption of cell membrane will occur rather than absorption over the cell membrane into the cell and transfer to the nucleus to interact with DNA.

 

Further information supporting the lack of genotoxic properties comes from the profiling of alkyl-diamines with varying chain length from C10 to C18, and including C18-unsaturated (Oleyl-diamine). (QSAR Toolbox v.4.1). There are no alerts are found for DNA interaction, protein interactions and no DNA alerts for Ames, MN and CA (OASIS v1.1).

Selecting in QSAR Toolbox all primary amines (from OECD HPV profile - total 1750 selected), and removing all compounds that are not discrete chemical and having other atoms besides carbon and nitrogen results to 306 relevant primary amines. From these there are 764 genotoxicity data points reported belonging to 68 of these subcategorized substances. Evaluation of all mutagenicity related data (608 data points of the 764), there was only one positive mutagenic result present, belonging to naphthylethylenediamine. This indicates a lack of mutagenic properties for the primary amines category of chemicals, to which the diamines also belong.

 

Information from QSARs on alkyl-diamine structures also showed no indication for mutagenicity:

- VEGA (Mutagenicity models CAESAR version 2.1.10; SarPy model, version 1.0.5-BETA): Predicts non-mutagenic, both with high reliability, but with the indication that compound could be out of the Applicability Domain of the model.

- DEREK (Derek Nexus: 3.0.1, Nexus: 1.5.0): Nothing to report on mutagenicity

- TOPKAT (Accelrys ADMET Toxicity Prediction (Extensible)) predicts non-mutagen, with high validity for oleyl-diamine, C18-diamine and C10-diamine (probability for mutagenicy of 0, 0.007 and 0.047 resp.).

- QSAR Toolbox v.3.0 contains series of QSAR for nodes under Human Health Hazards Genetic Toxicity that are all from Danish EPA DB. All eightIn vitroestimations and threein vivoestimations predicted negative genotoxicity for alkyl-diamine structures (in vitro:Ames test (Salmonella); UDS; DNA react. (Ashby fragments); Chrom. abber. (CHO); Mouse, COMET assay; HGPRT; Syrian hamster embryo cells; SCE;In vivo:Rodent, Dominant lethal; Drosophila sex-linked recessive lethal; Mouse micronucleus.).

 

Also the combined dataset of performed studies evaluating the genotoxicity hazards of alkyl-diamines substances indicate that alkyl-diamines do not have genotoxic properties. All of the available studies showed negative responses.

 

Available data on alkyl-diamines:

Test System or Species, Strain, Age, Number, and Sex of Animals	Biological Endpoint	S9	Chemical Form and Purity, vehicle	Dose	Results/Comments	Reference
Prokaryotic Systems
OECD 471, GLP S. typhimurium strainsTA 1535, TA 1537, TA 98 and TA 100	Increase in revertants due to mutations	+/-	Duomeen C Coco-diamine, 98.7% in DMSO	+S9: 0.33, 1.0, 3.3, 10.0, 33.3 µg/plate -S9: 3.3, 10.0, 33.3, 100 µg/plate	No mutagenic effects observed. Highest dose 1000 µg/plate was based on toxicity pre-test, Highest dose is selected to show slight toxicity.	Proprietary RCC Notox, 1990 031444
OECD 471, GLP S. typhimurium strainsTA98, TA100, TA1535, TA1537;Escherichia coliWP2 uvrA	Increase in revertants due to mutations	+/-	Genamin LAP 100D C12-diamine purity 100% in ethanol;	(0.16), 0.5, 1.6, 5, 16, 50, 160 µg/plate with and without S9	Precipitation at 5000 µg/plate No mutagenic effects observed under the test conditions. Toxic concentration observed for bacteria from 160 μg/ with and from 50 µg/plate without activation.	Proprietary Aventis, 2003 PT03-0028
OECD 471, GLP S. typhimurium strainsTA98, TA100, TA1535, TA1537;Escherichia coliWP2 uvrA	Increase in revertants due to mutations	+/-	Duomeen HT HT-diamine Purity np In ethanol	Exp.1: 0.316, 0.1, 3.16, 10.0, 31.6, 100, 316, 1000 and 2500 µL/plate Exp.2: 0.158, 0.50, 1.58, 5.0, 15.8, 50 , 158, 500 and 1580 µL/plate	Precipitation from 31.6 µg/plate without S9 and from 500 µg/plate with S9. No mutagenic effects observed under the test conditions. Toxic concentration observed for bacteria from 10 μg/plate without and from 100 µg/plate with activation.	Proprietary BSL, 2008 081561
OECD 471, GLP S. typhimurium strainsTA 1535, TA 1537, TA 1538, TA 98 and TA 100	Increase in revertants due to mutations	+/-	Dinoram SH HT-diamine 90% In ethanol	1, 5, 10, 25 and 50 mg /plate.	No mutagenic effects observed under the test conditions. Toxicity was observed from 50 µg/plate and higher	Proprietary CIT, 1986 2097 MMO
OECD 471, GLP S. typhimurium strainsTA98, TA100, TA1535, TA1537;Escherichia coliWP2 uvrA	Increase in revertants due to mutations	+/-	Duomeen OV Oleyl-diamine 92.3% In Ethanol	Exp.1: 0.00010, 0.000316, 0.00100, 0.00316, 0.0100, 0.0316 and 0.1 µL/plate Exp.2: 0.000050, 0.000158, 0.00050, 0.00158, 0.0050, 0.0158, 0.05 and 0.1 µL/plate	No precipitation of the test item was observed. No mutagenic effects observed under the test conditions. Toxic concentration observed for bacteria from 0.00316 µL/plate without and from 0.0316 µL/plate with activation.	Proprietary BSL, 2008 081576
Mammalian SystemsIn Vitro
OECD 473, GLP CHL (V79)	Chromosomal aberration	+/-	Genamin LAP 100D C12-diamine purity 100% in ethanol;	+S9: 0.16, 0.32, 0.64, 1.28 µg/ml -S9 3h: 0.16, 0.32, 0.64 µg/ml -S9 20h: 0.1, 0.2, 0.3 µg/ml	The test material was classified as “negative” for chromosomal aberrations, under the test conditions. Cell survival was reduced below 50 % in the highest evaluated concentrations. Higher dose levels were not evaluable because of an insufficient number of metaphases.	Proprietary Aventis, 2003 PT03-0029
OECD 473, GLP CHL (V79)	Chromosomal aberration	+/-	Duomeen OV Oleyl-diamine 92.3% In Ethanol	+S9: 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 6.5 and 8.0 µg/mL -S9 4h: 0.2, 0.4, 0.55, 0.7, 0.85, 1.0 and 1.2 µg/mL -S9 20h: 0.05, 0.1, 0.2, 0.4, 0.55, 0.7, 0.85, 1.0 and 1.2 µg/mL	The test material was classified as “negative” for chromosomal aberrations, under the test conditions. The lowest concentration producing cell toxicity was 4.0 mg/mL with metabolic activation and 1.0 (4h) and 0.4 (20h) mg/mL without metabolic activation.	Proprietary BSL, 2008 081575
OECD 476, GLP CHL (V79)	forward mutations (HPRT locus)	+/-	Duomeen OV Oleyl-diamine 92.3% In Ethanol	+S9: 0.05, 0.10, 0.25, 0.5, 1.0, 2.0, 3.0, 3.8, 4.0, 4.2, 5.0, 5.5, 6.0, 7.0 µg/mL -S9 4h: 0.350, 0.425, 0.500, 0.575, 0.650, 0.725, 0.800, 0.875 µg/mL -S9 20h: 0.1, 0.2, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 µg/mL	The test material was non-mutagenic in the HPRT locus using V79 cells of the Chinese hamster. Cytotoxicity was observed from 5.0 mg/mL with metabolic activation and 0.875 (4h) and 0.9 (20h) mg/mL without metabolic activation.	Proprietary BSL, 2010 092013
Mammalian SystemsIn Vivo
OECD474, GLP MouseSwiss Webster, adult, M and F	MN induction	na	Duomeen C Coco-diamine, in corn oil	Oral main study: 0, 31.3, 62.5 or 125 mg/kg/day; 5 animals/dose/sex + 125 mg additional 9 animals/sex	Does not increase frequency in micronuclei in mouse bone marrow PCE. Toxicity: 65% suppression PCE/RBC ratio at 125 mg	Proprietary. SRI, 1991 1924-C01-90

np = not provided; na = not applicable

 

References:

·        Aventis, 2003: Aventis Pharma, PT03-0028, 27-06-2003, GENAMIN LAP 100 D bacterial reverse mutation test.

·        Aventis, 2003: Aventis Pharma, PT03-0029, 16-07-2003, GENAMIN LAP 100 Din vitromammalian chromosome aberration test in V79 Chinese Hamster Cells.

·        RCC Notox, 1990: RCC Notox, 031444,, Evaluation of the mutagenic activity of Duomeen C in the Ames Salmonella/microsome test (with independent repeat).

·        SRI, 1991: SRI International, 1924-C01-90, 28-03-1991, Measurement of micronuclei in bone marrow erythrocytes of Swiss-Webster mice treated with Duomeen C.

·        BSL, 2008: BSL Bioservice, 081561, 22-09-2008, Reverse mutation assay using bacteria Salmonella typhimurium and Escherichia coli with N-(Hydrogenated tallow)-1,3-diaminopropane.

·        CIT, 1986: CIT, CIT, 2097 MMO, 21-05-1986, Dinoram SH - Study on the mutagenic properties in vitro in the Ames test.

·        BSL, 2008: BSL Bioservice, 081576, 23-09-2008, Reverse mutation assay using bacteria Salmonella typhimurium and Escherichia coli with N-Oleyl-1,3-diaminopropane

·        BSL, 2008: BSL Bioservice, 081575,, In vitro mammalian chromosome aberration test in Chinese hamster V79 cells with N-Oleyl.1,3.diaminopropane

·        BSL, 2010: BSL Bioservice, 092013, 29-04-2010, In vitro mammalian cell gene mutation test (HPRT-locus) in Chinese hamster V79 cells with N-Oleyl.1,3-diaminopropane

Justification for classification or non-classification

Based on the three negative in-vitro tests for genotoxicity, N-Oleyl-1,3-diaminopropane, CAS 7173-62-8, does not require classification as a mutagen according to the European Union CLP/GHS criteria.

Also further property data for Oleyl-diamine indicate that genotoxic properties are rather unlikely.