Amines, tri-C8-10-alkyl

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

OECD guideline study conducted in compliance with GLP regulations, with acceptable restrictions (neither E. coli strain WP2 uvrA nor S. typhimurium strain TA102 were employed in this study for the detection of oxidising mutagens and cross-linking agents)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

(adopted 1983)

Deviations:

yes

Remarks:

(neither E. coli strain WP2 uvrA pKM101a nor S. typhimurium strain TA102 were employed in this study for the detection of oxidising mutagens and cross-linking agents; 2-aminoanthracene used as sole indicator of the efficacy of the S9 mix)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Amines, tri-C8-10-alkyl

EC Number:

272-347-8

EC Name:

Amines, tri-C8-10-alkyl

Cas Number:

68814-95-9

Molecular formula:

Unspecified

IUPAC Name:

Amine, tri(n-C8,C10-alkyl)

Test material form:

other: liquid

Details on test material:

- Name of test material (as cited in study report): Alamine 336
- Physical state: liquid, colourless
- Analytical purity: 100 % (active substance)
- Lot/batch No.: 5J167
- Expiration date of the lot/batch: February 2000
- Stability under test conditions: stable for 12 months
- Storage condition of test material: at room temperature
- SAT-No.: 990409

Method

Target gene:

his operon

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537, TA 1538

Metabolic activation:

with and without

Metabolic activation system:

Phenobarbital/B-Naphthoflavone induced rat liver S-9 mix

Test concentrations with justification for top dose:

1st and 2nd assay: 8, 40, 200, 1000 and 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Tween 80/bidest. water
- Justification for choice of solvent/vehicle: The suspension medium was chosen according to the solubility properties tested preliminary before start of the study.

Controls

Untreated negative controls:

yes

Remarks:

untreated cell suspension

Negative solvent / vehicle controls:

yes

Remarks:

Tween 80/bidest. water

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

sodium azide

other: 4-nitro-o-phenylenediamine, 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: 12 hours

NUMBER OF REPLICATIONS: triplicate plates

DETERMINATION OF CYTOTOXICITY
- Method: background lawn

DETAILS ON CONTROLS:
- Negative controls: suspension medium Tween 80/bidist. water and the untreated fresh cell suspensions
- Positive controls:
- Without metabolic activation:
sodium azide (2 µg/plate) for TA 100 and TA 1535;
9-aminoacridine (80 µg/plate) for TA 1537
4-nitro-o-phenylendiamine (40 µg/plate) for TA 98 and TA 1538
- With metabolic activation:
2-aminoanthracene in all strains (2.5 µg/plate for TA 1535, TA 1537; 5.0 µg/plate for TA 100, TA 98, TA 1538)
- Sterility controls:
The test batches containing S9-mix were each controlled for sterility by adding 0.5 mL of S9-mix fraction to untreated agar plates.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

RESULTS:

 

Experiment 1
Treatment groups	[µg/plate]	Strains
		TA 100							TA 1535							TA 1537				TA 1538				TA 98
		-	SD		+		SD		-	SD		+		SD		-	SD	+	SD	-	SD	+	SD	-	SD	+	SD
medium	-	108	6		125		15		9	3		18		12		16	5	10	0	10	4	11	6	19	8	32	2
solvent	-	129	15		117		14		10	1		13		1		17	5	11	3	8	1	12	7	22	6	25	4
test substance	8	121	18		137		17		10	2		11		3		14	3	20	2	10	3	9	6	29	8	22	3
	40	121	15		134		16		14	5		14		4		9	2	13	4	15	5	11	2	24	3	27	2
	200	115	12		113		17		7	2		11		3		14	1	4	3	13	3	15	3	23	8	31	5
	1000	113	11		98		9		9	1		11		6		13	3	12	3	8	3	11	3	20	4	29	5
	5000	106	8		87		16		11	4		7		4		5	1	5	2	11	5	14	1	14	3	22	7
positive controls	A	736	40		302		6		542	8		222		13		906	59	463	56	2421	54	1784	75	1718	100	1381	166
	B	148	13		2990		18		11	1		194		35		15	6	181	15	17	3	1992	122	19	3	2125	110
Experiment 2
Treatment groups	[µg/plate]	Strains
		TA 100							TA 1535							TA 1537				TA 1538				TA 98
		-		SD		+		SD	-		SD		+		SD	-	SD	+	SD	-	SD	+	SD	-	SD	+	SD
medium	-	103		12		116		15	8		5		10		2	14	5	20	7	13	7	18	8	23	3	20	6
solvent	-	111		15		96		16	24		2		9		3	10	2	12	1	16	7	12	2	26	4	24	2
test substance	8	115		6		117		15	14		7		16		1	10	1	15	5	10	3	17	6	23	0	23	6
	40	111		3		125		24	12		4		15		4	14	3	10	4	13	1	14	3	23	2	28	2
	200	106		11		119		23	15		2		13		5	7	1	8	3	13	4	13	3	21	2	30	5
	1000	81		11		82		3	12		5		10		1	7	2	9	5	12	4	12	3	13	2	23	2
	5000	82		11		86		9	152		2		12		6	8	3	10	5	8	2	11	1	13	1	25	2
positive controls	A	764		19		165		22	615		21		76		9	409	120	112	28	2387	235	1227	297	1587	21	872	96
	B	118		9		943		91	27		9		115		25	18	4	78	8	16	1	951	123	27	6	727	75

-: without metabolic activation

+: with metabolic activation

SD: standard deviation

Positive controls:         A:        TA 1535, TA 100:                     sodium azide

                                           TA 1537:                                9-aminoacridine

                                           TA 1538, TA 98:            4-nitro-o-phenylenediamine

                                B :       TA 1535, TA 1537:                    2-aminoanthracene

                                           TA 100, TA 1538, TA 98:            2-aminoanthracene

Applicant's summary and conclusion

Conclusions:

Interpretation of results: negative

Amines, tri-C8-10-alkyl is considered not to be mutagenic in this bacterial mutagenicity test in vitro.

Executive summary:

The test item with the batch number 5J167 was tested in the Salmonella typhimurium Reverse Mutation Assay for the induction of reverse mutations in a bacterial test system. The assay was performed with the Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and TA 1538 in two independent experiments, both with and without metabolic activation by S9-mix according to OECD 471 (BASF SE, 1999, 9901542).

Solutions of the test substance were prepared in Tween 80/bidist. water and diluted with Tween 80/bidist. water just before use. The following concentrations were tested:

1st and 2nd test: 8, 40, 200, 1000 and 5000 µg/plate

The direct plate incorporation assay was utilized. The following results were obtained:

Toxic effects were not noted. No enhanced revertant rates compared to concurrent negative controls were observed in all tested strains, neither in the presence nor in the absence of metabolic activation.

Therefore the test substance did not induce reverse mutations in the tested strains of Salmonella typhimurium in this bacterial mutagenicity test.

In conclusion, it can be stated that in the study described and under the experimental conditions reported, the test substance did not induce gene mutations in Salmonella typhimurium strains.

Subsequently, Amines, tri-C8-10-alkyl is considered not to be mutagenic in this

bacterial mutagenicity test in vitro.