Hexadecylamine

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Based on Ames test it can be stated that the test item is not mutagenic in the strains TA 100, TA 1535, TA 97 and TA 98 of Salmonella typhimurium with and without metabolic activation (reference 7.6.1 -2].

Additionaly, the read-across item number 2 is not mutagenic in the strains TA 100, TA 1535, TA 1537, TA 1538, TA 98 of Salmonella typhimurium and Escherichia coli WP2uvrA (reference 7.6.1 -2).

Overall no mutagenic effects were described.

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

Data from summary document with only 4 strains; acceptable because data consistent with other data in subcategory. The endpoint has been adequately characterized. (American Chemistry Council, Fatty Nitrogen Derivatives Panel, Amines Task Group).

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Target gene:

Histidine (S. typhimurium)

Species / strain / cell type:

S. typhimurium TA 97

Species / strain / cell type:

S. typhimurium TA 98

Species / strain / cell type:

S. typhimurium TA 100

Species / strain / cell type:

S. typhimurium TA 1535

Metabolic activation:

with and without

Metabolic activation system:

The S-9 (9,000g supernatant) fractions of Aroclor 1254-induced, male Sprague-Dawley rat and male Syrian hamster livers were prepared as described previously [Haworth et al, 1983]. The S-9 mixes were prepared immediately prior to use and contained either 10% or 30% S-9. The test substance was tested in the absence of metabolic activation and with rat and hamster S-9 fractions.

Test concentrations with justification for top dose:

0.3, 1.0, 3, 10, 33, 100 µg/plate based on toxicity pretest

Vehicle / solvent:

DSMO

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

sodium azide

other: 4-nitro-o-phenylenediamine: TA98 (without S9); 2-aminoan-thracene: all strains (with S9)

Details on test system and experimental conditions:

The preincubation assay was performed as described previously [Haworth et al., 1983], with some differences, as described below. The test chemical (0.05 ml), Salmonella culture (0.10 ml), and S-9 mix or buffer (0.50 ml) were incubated at 37°C, without shaking, for 20 min. Chemicals known or suspected to be volatile were incubated in capped tubes. The top agar was added and the contents of the tubes were mixed and poured onto the surface of petri dishes containing Vogel-Bonner medium [Vogel and Bonner, 1956]. The histidine-independent (his+) colonies arising on these plates were counted following two days incubation at 37°C. Plates were machine counted (New Brunswick, Edison, NJ; Artek, Farmingdale, NY) unless precipitate was present which interfered with the count, or the color of the test chemical on the plate reduced the contrast between the colonies and the background agar. At the discretion of the investigators, plates with low numbers of colonies were counted by hand.
Variations in the protocol among the tested chemicals reflect the evolution of the protocol originally described by Haworth et al. [1983].
The test substance was tested initially in a toxicity assay to determine the appropriate dose range for the mutagenicity assay. The toxicity assay was performed using TA100 or the sytem developed by Waleh et al. [1982]. Toxic concentrations were those that produced a decrease in the number of his+ colonies, or a clearing in the density of the background lawn, or both. The test substance was tested initially at half-log dose intervals up to a dose that elicited toxicity, or to a dose immediately below one which was toxic in the preliminary toxicity test. Subsequent trials occasionally used narrower dose increments and may not have included doses in the toxic range. Concurrent solvent and positive controls were run. The positive controls in the absence of metabolic activation were sodium azide (TA1535 and TA100), 9-aminoacridine (TA97 and TA1537), and 4-nitro-o-phenylenediamine (TA98). The positive control for metabolic activation with all strains was 2-aminoan-thracene.

Evaluation criteria:

The data were evaluated as described previously [Zeiger et al., 1987]. Evaluations were made at both the individual trial and overall chemical levels. Individual trials were judged mutagenic (+), weakly mutagenic (+W), questionable (?), or nonmutagenic (-), depending on the magnitude of the increase of his+ revertants, and the shape of the dose-response. A trial was considered questionable (?) if the dose-response was judged insufficiently high to support a call of " +W," if only a single dose was elevated over the control, or if the increase seen was not dose related. The distinctions between a questionable mutagenic response and a nonmutagenic or weak mutagenic response and between a weak mutagenic response and mutagenic response are highly subjective. It was not necessary for a response to reach twofold over background for a chemical to be judged mutagenic. A chemical was judged mutagenic (+) or weakly mutagenic (+W) if it produced a reproducible dose-related response over the solvent control in replicate trials. A chemical was judged questionable (?) if the results of individual trials were not reproducible, if increases in his4" revertants did not meet the criteria for a "+W" response, or if only single doses produced increases in his +revertants in repeat trials. Chemicals were judged nonmutagenic (-) if they did not meet the criteria for a mutagenic or questionable response. The chemicals were decoded by the chemical repository only after a determination had been made regarding their mutagenicity or nonmutagenicity.

Key result

Species / strain:

S. typhimurium, other: TA 97, 98, 100, 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 100 µg/plate

Vehicle controls validity:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

No historical control data is available, but the control plates without activation were comparable to that described in literature.

Table 1: Detailed results for TA100 and TA1535

Dose	TA100										TA1535
	NA		10% HLI		30% HLI		10% RLI		30% RLI		NA		10% HLI		30% HLI		10% RLI		30% RLI
µg/PLATE	mean	SD	mean	SD	mean	SD	mean	SD	mean	SD	mean	SD	mean	SD	mean	SD	mean	SD	mean	SD
0.000	120	3.1	108	1.3	127	7.9	118	4.4	142	1.9	10	3.4	6	2.2	16	1.9	6	0.9	9	2.6
0.300	106	3.1									15	4.3
1.000	93	5.5	113	28.5	131	2.9	130	33.5	127	17.1	17	2.5	12	5.8	13	0.7	8	2.8	17	4.0
3.000	63	4.2	110	17.3	117	10.5	119	8.5	110	9.9	11	3.2	6	0.7	7	1.3	7	1.5	7	1.7
10.000	73	6.8	121	6.9	131	5.2	165	15.0	113	12.7	15	0.9	7	1.0	9	3.0	7	1.0	12	1.9
33.000	69	9.8	116	15.0	124	9.0	161	12.5	114	9.3	12	2.3	10	1.5	9	0.3	8	3.2	8	1.9
100.000			78	10.0	112	7.2	97	5.5	134	13.7			8	3.0	5	1.3	4	0.9	9	0.9
POS	274	5.2	2127	115.0	748	9.6	1380	84.1	352	4.6	186	3.0	635	8.0	534	33.8	460	24.6	110	11.1

Abbreviations:

POS: positive control

NA: not activated

HLI: Aroclor 1254-induced hamster liver S-9

RLI: Aroclor 1254-induced rat liver S-9

 

Table 2: Detailed results of TA97 and TA98

Dose	TA97										TA98
	NA		10% HLI		30% HLI		10% RLI		30% RLI		NA		10% HLI		30% HLI		10% RLI		30% RLI
µg/PLATE	mean	SD	mean	SD	mean	SD	mean	SD	mean	SD	mean	SD	mean	SD	mean	SD	mean	SD	mean	SD
0.000	153	2.6	216	5.2	173	9.2	216	15.6	224	7.6	12	3.7	45	5.7	22	2.5	34	1.7	21	0.9
0.300	155	2.6									11	1.3
1.000	140	9.8	255	19.7	199	30.0	255	11.8	234	12.2	14	3.2	39	6.9	22	2.3	35	3.5	28	0.9
3.000	126	9.0	230	3.3	183	15.9	235	19.3	191	13.3	7	0.6	35	11.2	21	1.5	28	1.5	25	2.8
10.000	114	6.1	255	11.3	191	16.3	286	11.3	207	14.3	10	1.5	36	2.0	15	2.0	31	4.7	15	2.5
33.000	101	4.0	225	9.0	182	12.3	265	4.2	204	11.3	9	2.3	32	2.0	13	3.4	26	0.9	19	4.0
100.000			195	14.2	192	5.2	212	15.0	203	9.6			21	4.1	13	3.5	22	6.0	23	3.5
POS	106	141.6	1121	53.1	1007	50.1	1767	80.4	579	7.0	740	9.8	1207	133.9	234	7.0	830	110.3	151	3.7

Abbreviations:

POS: positive control

NA: not activated

HLI: Aroclor 1254-induced hamster liver S-9

RLI: Aroclor 1254-induced rat liver S-9

Conclusions:

Based on the test results it can be stated that the test item is not mutagenic in the strains TA 100, TA 1535, TA 97 and TA 98 of Salmonella typhimurium with and without metabolic activation.

Executive summary:

The test item was tested for mutagenicity with the strains TA 100, TA 1535, TA97 and TA 98 of Salmonella typhimurium. The mutagenicity study was conducted in the absence and in the presence of a metabolizing system derived from rat and hamster liver homogenate. Control plates without mutagen showed that the number of spontaneous revertant colonies was comparable to that described in literatur. All the positive control compounds gave the expected increase in the number of revertant colonies. In the cytotoxicity trials, the test compound proved to be toxic to most of the bacterial strains at 100 microgram/plate. These tests demonstrated that in the presence or absence of the metabolic activation system the test compound did not show a dose dependent increase in the number of revertants in any of the bacterial strains. Summarizing, it can be stated that the test item is not mutagenic in these bacterial test systems either with or without exogenous metabolic activation at the dose levels investigated.

Reference 2

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1988

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Target gene:

Histidine (S. typhimurium)
Tryptophan (E. coli)

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain / cell type:

S. typhimurium TA 1538

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

4, 20, 100 500, 2500, 10000 µg/plate (first experiment)
4, 20, 100 500, 2500, 5000 µg/plate (second experiment)

Vehicle / solvent:

- Vehicle/solvent used: ethanol

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: - S9: Sodium azide (TA 100, TA 1535), 9-Aminoacridine (TA 1537), 2-Nitrofluorene (TA98, TA 1538), N-Methyl-N-nitro-N-nitrosoguanidine (WP2 uvrA); +S9: Benzo(a)pyrene, 2-Aminoanthracene (all strains)

Details on test system and experimental conditions:

NUMBER OF REPLICATIONS:
- Number of cultures per concentration: triplicate
- Number of independent experiments: 2

METHOD OF TREATMENT/ EXPOSURE:
- Test substance added in agar

TREATMENT AND HARVEST SCHEDULE:
- Duration of treatment: 72 h

FOR GENE MUTATION:
- Method used: agar
- Number of cells seeded and method to enumerate numbers of viable and mutants cells: not specified

METHODS FOR MEASUREMENT OF CYTOTOXICITY
- Method: background growth inhibition

Key result

Species / strain:

other: all strains tested

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

The test compound proved to be toxic to most of the bacterial strains at doses of 100 µg/plate. Thinning of the bacterial lawn and a reduction in the number of colonies have been observed at this dose. Visible precipitation of the test compound on the plates has been observed at 500 µg/plate. The test compound did not cause a significant increase in the number of revertant colonies with any of the tester strains either in the absence or presence of S-9 Mix. No dose dependent effect was obtained. No unforeseen circumstances were observed which have affected the quality and integrity of this study.

Table 1: Mean number of revertants without S9 mix (experiment 1)

Test substance	Dose/Plate (µg)	TA 98	TA 100	TA 1535	TA 1537	TA 1538	WP2uvrA
Test substance	0	20	149	13	10	11	60
	4	23	154	13	9	13	60
	20	20	146	13	13	13	52
	100	12	58	10	6	7	34
	500	3p	20p	8p	1p	3p	21p
	2500	1p	8p	5p	2p	2p	17p
	10000	0p	14p	4p	0p	0p	25p
Soldium-azide	1		424	274
9-Aminoacridine	50				147
2-Nitrofluorene	2.5	336				435
MNNG	2.5						253

p: visible precipitation of the test compound on the plates

Table 2: Mean number of revertants with S9 mix (experiment 1)

Test substance	Dose/Plate (µg)	TA 98	TA 100	TA 1535	TA 1537	TA 1538	WP2uvrA
Test substance	0	37	153	15	10	17	69
	4	28	173	15	8	15	63
	20	33	172	17	6	16	55
	100	27	156	15	9	15	47
	500	13p	65p	12p	5p	8p	34p
	2500	1p	53p	8p	2p	4p	35p
	10000	4p	73p	10p	5p	7p	35p
2-Aminoanthracen	0.5 - 10	507	549	90	87	428	496
Benzo[a]pyrene	10	547	902	18	100	208	98

p: visible precipitation of the test compound on the plates

 

Table 3: Mean number of revertants without S9 mix (experiment 2)

Test substance	Dose/Plate (µg)	TA 98	TA 100	TA 1535	TA 1537	TA 1538	WP2uvrA
Test substance	0	25	134	11	7	10	48
	4	26	153	13	7	12	53
	20	22	128	12	6	10	47
	100	16	38	10	6	10	40
	500	6p	9p	8p	2p	6p	26p
	2500	3p	3p	1p	1p	3p	20p
	5000	0p	0p	0p	0p	0p	22p
Soldium-azide	1		374	297
9-Aminoacridine	50				126
2-Nitrofluorene	2.5	402				376
MNNG	2.5						216

p: visible precipitation of the test compound on the plates

 

Table 4: Mean number of revertants with S9 mix (experiment 2)

Test substance	Dose/Plate (µg)	TA 98	TA 100	TA 1535	TA 1537	TA 1538	WP2uvrA
Test substance	0	31	175	12	9	15	59
	4	29	178	14	9	13	56
	20	27	182	17	8	14	49
	100	18	105	15	7	13	43
	500	8p	41p	12p	5p	7p	38p
	2500	2p	23p	10p	3p	5p	26p
	10000	0p	13p	7p	2p	2p	28p
2-Aminoanthracen	0.5 - 10	487	610	102	106	348	446
Benzo[a]pyrene	10	596	918	17	92	138	93

p: visible precipitation of the test compound on the plates

Conclusions:

Based on the test results it can be stated that octadecan-1 -amine (CAS 124 -30 -1) is not mutagenic in the strains TA 100, TA 1535, TA 1537, TA 1538, TA 98 of Salmonella typhimurium and Escherichia coli WP2uvrA.

Executive summary:

The test substance was tested for mutagenicity with the strains TA 100, TA 1535, TA 1537, TA 1538, TA 98 and Escherichia coli WP2uvrA. The mutagenicity studies were conducted in the absence and in the presence of a metabolizing system derived from rat liver homogenate. A dose range of 6 different doses from 4 microgram/plate to 5000 microgram/plate was used. Control plates without mutagen showed that the number of spontaneous revertant colonies was similar to that described in literatur and comparable to historical controls. All the positive control compounds gave the expected increase in the number of revertant colonies. In the cytotoxicity trials, the test compound proved to be toxic to most of the bacterial strains at 100 microgram/plate. 5000 microgram/plate was chosen as top dose level for the mutagenicity studies. These tests demonstrated that in the absence of the metabolic activation system the test compound did not show a dose dependent increase in the number of revertants in any of the bacterial strains. Also in the presence of a metabolic activation system, treatment of the cells with the test item did not result in relevant increases in the number of revertant colonies. Summarizing, it can be stated that the test item is not mutagenic in these bacterial test systems either with or without exogenous metabolic activation at the dose levels investigated.

Reference 3

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

Please refer to Section 13.

Reason / purpose for cross-reference:

read-across source

Key result

Species / strain:

other: all strains

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

The test item was tested for mutagenicity with the strains TA 100, TA 1535, TA97 and TA 98 of Salmonella typhimurium. The mutagenicity study was conducted in the absence and in the presence of a metabolizing system derived from rat and hamster liver homogenate. Control plates without mutagen showed that the number of spontaneous revertant colonies was comparable to that described in literatur. All the positive control compounds gave the expected increase in the number of revertant colonies. In the cytotoxicity trials, the test compound proved to be toxic to most of the bacterial strains at 100 microgram/plate. These tests demonstrated that in the presence or absence of the metabolic activation system the test compound did not show a dose dependent increase in the number of revertants in any of the bacterial strains. Summarizing, it can be stated that the test item is not mutagenic in these bacterial test systems either with or without exogenous metabolic activation at the dose levels investigated.

Additionaly the read-across substance number 2 was tested for mutagenicity with the strains TA 100, TA 1535, TA 1537, TA 1538, TA 98 and Escherichia coli WP2uvrA. The mutagenicity studies were conducted in the absence and in the presence of a metabolizing system derived from rat liver homogenate. A dose range of 6 different doses from 4 microgram/plate to 5000 microgram/plate was used. Control plates without mutagen showed that the number of spontaneous revertant colonies was similar to that described in literatur and comparable to historical controls. All the positive control compounds gave the expected increase in the number of revertant colonies. In the cytotoxicity trials, the test compound proved to be toxic to most of the bacterial strains at 100 microgram/plate. 5000 microgram/plate was chosen as top dose level for the mutagenicity studies. These tests demonstrated that in the absence of the metabolic activation system the test compound did not show a dose dependent increase in the number of revertants in any of the bacterial strains. Also in the presence of a metabolic activation system, treatment of the cells with teh test item did not result in relevant increases in the number of revertant colonies. Summarizing, it can be stated that the test item is not mutagenic in these bacterial test systems either with or without exogenous metabolic activation at the dose levels investigated.

For the target substance and the second source substance the Derek Nexus Report was analyzed. For both substances mutagenicity in vitro in bacterium is inactive, no misclassification or unclassified features were detected.

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008
The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. Based on available data on genetic toxicity, the test item is not classified and labelled as genetic toxic according to Regulation (EC) No 1272/2008 (CLP), as amended for the fifteenth time in Regulation (EU) No 2020/1182.