Amines, C12-14-branched alkyl, dodecylbenzenesulfonates (1:1)

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

13 February - 06 November 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study conducted in accordance with international guidelines and in accordance with GLP. All guideline validity criteria were met.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Amines, C12-14-branched alkyl, octylbenzenesulfonates (1:1)

Molecular formula:

Amines, C12-branched alkyl, octylbenzenesulfonates: C27H51NO3S Amines, C13-branched alkyl, octylbenzenesulfonates: C28H53NO3S Amines, C14-branched alkyl, octylbenzenesulfonates: C29H55NO3S

IUPAC Name:

Amines, C12-14-branched alkyl, octylbenzenesulfonates (1:1)

Constituent 2

Reference substance name:

Amines, C12-14-branched alkyl, nonylbenzenesulfonates (1:1)

Molecular formula:

Amines, C12-branched alkyl, nonylbenzenesulfonates: C28H53NO3S Amines, C13-branched alkyl, nonylbenzenesulfonates: C29H55NO3S Amines, C14-branched alkyl, nonylbenzenesulfonates: C30H57NO3S

IUPAC Name:

Amines, C12-14-branched alkyl, nonylbenzenesulfonates (1:1)

Constituent 3

Reference substance name:

Amines, C12-14-branched alkyl, decylbenzenesulfonates (1:1)

Molecular formula:

Amines, C12-branched alkyl, decylbenzenesulfonates: C29H55NO3S Amines, C13-branched alkyl, decylbenzenesulfonates: C30H57NO3S Amines, C14-branched alkyl, decylbenzenesulfonates: C31H59NO3S

IUPAC Name:

Amines, C12-14-branched alkyl, decylbenzenesulfonates (1:1)

Constituent 4

Reference substance name:

Amines, C12-C14-branched alkyl, benzenesulfonates C12-C15 (1:1)

Molecular formula:

Amines, C12-branched alkyl, benzenesulfonates C12 (1:1) C31H59NO3S Amines, C12-branched alkyl, benzenesulfonates C13 (1:1) C32H61NO3S Amines, C12-branched alkyl, benzenesulfonates C14 (1:1) C33H63NO3S Amines, C12-branched alkyl, benzenesulfonates C15 (1:1) C34H65NO3S Amines, C13-branched alkyl, benzenesulfonates C12 (1:1) C32H61NO3S Amines, C13-branched alkyl, benzenesulfonates C13 (1:1) C33H63NO3S Amines, C13-branched alkyl, benzenesulfonates C14 (1:1) C34H65NO3S Amines, C13-branched alkyl, benzenesulfonates C15 (1:1) C35H67NO3S Amines, C14-branched alkyl, benzenesulfonates C12 (1:1) C33H63NO3S Amines, C14-branched alkyl, benzenesulfonates C13 (1:1) C34H65NO3S Amines, C14-branched alkyl, benzenesulfonates C14 (1:1) C35H67NO3S Amines, C14-branched alkyl, benzenesulfonates C15 (1:1) C36H69NO3S

IUPAC Name:

Amines, C12-C14-branched alkyl, benzenesulfonates C12-C15 (1:1)

Constituent 5

Reference substance name:

Amines, C12-14-branched alkyl, hexadecylbenzenesulfonates (1:1)

Molecular formula:

Amines, C12-branched alkyl, hexadecylbenzenesulfonates (1:1): C35H67NO3S Amines, C13-branched alkyl, hexadecylbenzenesulfonates (1:1): C36H69NO3S Amines, C14-branched alkyl, hexadecylbenzenesulfonates (1:1): C37H71NO3S

IUPAC Name:

Amines, C12-14-branched alkyl, hexadecylbenzenesulfonates (1:1)

Constituent 6

Reference substance name:

Residual Amines, C12-14-tert-alkyl

IUPAC Name:

Residual Amines, C12-14-tert-alkyl

Test material form:

liquid: viscous

Details on test material:

Storage: Room Temperature

Specific details on test material used for the study:

RADIOLABELLING INFORMATION (if applicable)
- Radiochemical purity: N/A
- Specific activity: N/A
- Locations of the label: N/A
- Expiration date of radiochemical substance: N/A

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: 15-25 ºC, protected from light
- Stability under test conditions: Assumed stable
- Solubility and stability of the test substance in the solvent/vehicle: Soluble at 573.6 mg/mL in DMSO
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: No

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Test article stock solutions were prepared by formulating the test article under subdued lighting in DMSO with the aid of vortex mixing and warming at 37 ̊C, to give the maximum required treatment concentration. Subsequent dilutions were made using DMSO.
- Preliminary purification step (if any): N/A
- Final dilution of a dissolved solid, stock liquid or gel: 50 mg/mL prepared as the solution for the highest tested concentration. Subsequent dilutions were made using DMSO.
- Final preparation of a solid: N/A

FORM AS APPLIED IN THE TEST (if different from that of starting material) : Appled as a liquid

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable) : N/A

OTHER SPECIFICS: N/A

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

A maximum concentration of 5000 μg/plate was selected for Experiment 1, to ensure that the initial treatments were performed up to this maximum recommended concentration according to current regulatory guidelines.

Experiment 1 (with and without S9 mix): 5000, 1600, 500, 160, 50, 16, 5 µg/plate
Experiment 2 (without S9 mix): 1600, 800, 400, 200, 100, 50 µg/plate
Experiment 2 (with S9 mix): 1600, 800, 400, 200, 100, 50 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: Maron, D., Katzenellenbogen, J., and Ames, B.N. (1981). Compatibility of Organic Solvents
with the Salmonella/Microsome Test. Mutation Res., 88, 343-350.

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO; (Experiment 1: batch number STBG36578V, supplied by Sigma-Aldrich Chemical Company, no expiry date specified by the supplier; Experiment 2: batch number STBG4158V, supplied by Sigma-Aldrich Chemical Company, no expiry date specified by the supplier

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

benzo(a)pyrene

mitomycin C

other: 2-aminoanthracene (AAN)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)
- Cell density at seeding (if applicable): 10E8 - 10E9 cells/mL

DURATION
- Preincubation period: 20 mins (Experiment 2, with S9 mix only)
- Exposure duration: 2-3 days
- Expression time (cells in growth medium): 2-3 days
- Selection time (if incubation with a selection agent): N/A
- Fixation time (start of exposure up to fixation or harvest of cells): N/A

SELECTION AGENT (mutation assays): N/A

SPINDLE INHIBITOR (cytogenetic assays): N/A

STAIN (for cytogenetic assays): N/A

NUMBER OF REPLICATIONS: 3

METHODS OF SLIDE PREPARATION AND STAINING TECHNIQUE USED: N/A

NUMBER OF CELLS EVALUATED: N/A

NUMBER OF METAPHASE SPREADS ANALYSED PER DOSE (if in vitro cytogenicity study in mammalian cells): N/A

CRITERIA FOR MICRONUCLEUS IDENTIFICATION: N/A

DETERMINATION OF CYTOTOXICITY
- Method: thinning/ absence of bacteria lawn

OTHER EXAMINATIONS:
- Determination of polyploidy: N/A
- Determination of endoreplication: N/A
- Methods, such as kinetochore antibody binding, to characterize whether micronuclei contain whole or fragmented chromosomes (if applicable): N/A

- OTHER: N/A

Rationale for test conditions:

The study was based on the in vitro technique described by Ames et al (1975), Maron and Ames (1983) and Mortelmans and Zeiger (2000), in which mutagenic effects are determined by exposing mutant strains of Salmonella typhimurium to various concentrations of the test item. These strains have a deleted excision repair mechanism which makes them more sensitive to various mutagens and they will not grow on media which does not contain histidine. When large numbers of these organisms are exposed to a mutagen, reverse mutation to the original histidine independent form takes place. These are readily detectable due to their ability to grow on a histidine deficient medium. Using these strains of Salmonella typhimurium revertants may be produced after exposure to a chemical mutagen, which have arisen as a result of a base-pair substitution in the genetic material (miscoding) or as a frameshift mutation in which genetic material is either added or deleted

Evaluation criteria:

For valid data, the test article was considered to be mutagenic if:

1. A concentration related increase in revertant numbers was ≥1.5-fold (in strain TA102), ≥2-fold (in strains TA98 or TA100) or ≥3-fold (in strains TA1535 or TA1537) the concurrent vehicle control values.
2. Any observed response was reproducible under the same treatment conditions.

The test article was considered positive in this assay if both of the above criteria were met.
The test article was considered negative in this assay if neither of the above criteria were met.

Statistics:

The statistical significance of results was analysed using Dunnett’s test to aid in the evaluation of potential positive responses.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Any other information on results incl. tables

Table 1       Raw plate counts and calculated mutagenicity data, experiment 1 (- S9 mix)

 

Strain	Compound	Conc. level (µg/plate)	Mean revertants per plate	SD	Fold increase	Dunnett’s T-value	Statistic. sig.	Revertant numbers/plate
TA98	DMSO	-	22.0	1.7	-	-	-	21, 24, 21
	Test item	5	20.0	1.0	0.9	-0.36	NS	21, 19, 20
		16	21.7	6.0	1.0	-0.10	NS	28, 16, 21
		50	14.3	4.7	0.7	-1.55	NS	16, 9, 18
		160	19.0	7.0	0.9	-0.62	NS	27, 14, 16
		500	15.0	10.4	0.7	-1.57	NS	9, 9, 27
		1600	2.0	1.7	0.1	-5.51	NS	1S, 1MS, 4S
		5000	2.0	1.7	0.1	-5.81	NS	3SP, 3SP, 0SP
	2NF	5	419.0	97.3	19.0	-	-	527, 338, 392
TA100	DMSO	-	117.3	6.8	-	-	NS	112, 115, 125
	Test item	5	96.0	21.0	0.8	-1.62	NS	120, 87, 81
		16	95.3	19.2	0.8	-1.66	NS	116, 92, 78
		50	84.3	7.8	0.7	-2.50	NS	78, 93, 82
		160	101.0	12.5	0.9	-1.20	NS	97, 115, 91
		500	60.0	14.9	0.5	-4.72	NS	54S, 49S, 77S
		1600	2.0	1.7	0.0	-14.64	NS	3V, 0V, 3V
		5000	-	-	-	-	-	-PT, -PT, -PT
	NaN3	2	450.0	20.4	3.8	-	-	515, 554, 507
TA1535	DMSO	-	8.7	3.2	-	-	-	10, 5, 11
	Test item	5	9.7	0.6	1.1	0.39	NS	10, 10, 9
		16	14.3	7.6	1.7	1.54	NS	11, 23, 9
		50	10.3	6.8	1.2	0.38	NS	5, 18, 8
		160	11.3	3.2	1.3	0.85	NS	9, 10, 15
		500	9.3	4.0	1.1	0.19	NS	5, 10, 13
		1600	1.7	1.2	0.2	-3.20	NS	1S, 3S, 1S
		5000	3.3	0.6	0.4	-2.09	NS	4SP, 3SP, 3SP
	NaN3	2	525.3	25.1	60.6	-	-	515, 554, 507
TA1537	DMSO	-	6.0	2.6	-	-	-	5, 9, 4
	Test item	5	7.0	2.6	1.2	0.31	NS	4, 8, 9
		16	9.0	7.5	1.5	0.49	NS	1, 10, 6
		50	5.0	2.6	0.8	-0.36	NS	4, 3, 8
		160	7.0	2.6	1.2	0.31	NS	4, 9, 8
		500	4.0	0.0	0.7	-0.65	NS	4, 4, 4
		1600	-	-	-	-	-	-T, -T, -T
		5000	-	-	-	-	-	-PT, -PT, -PT
	AAC	50	218.3	109.5	36.4	-	-	326, 222, 107
TA102	DMSO	-	246.7	45.7	-	-	-	299, 226, 215
	Test item	5	229.0	10.6	0.9	-0.78	NS	237, 233, 217
		16	257.0	41.6	1.0	0.49	NS	231, 305, 235
		50	233.3	11.0	0.9	-0.57	NS	221, 237, 242
		160	169.0	5.0	0.7	-3.88	NS	164S, 174S, 169S
		500	58.3	8.5	0.2	-11.71	NS	67S, 50S, 58S
		1600	-	-	-	-	-	-T, -T, -T
		5000	-	-	-	-	-	-PT, -PT, -PT
	MMC	0.2	696.3	174.4	2.8	-	-	657, 545,887

P: precipitation of test item observed

S: slight thinning of background bacterial lawn

T: toxic, no revertant colonies

V: very thin background bacterial lawn

Table 2       Raw plate counts and calculated mutagenicity data, experiment 1 (+ S9 mix)

 

Strain	Compound	Conc. level (µg/plate)	Mean revertants per plate	SD	Fold increase	Dunnett’s T-value	Statistic. sig.	Revertant numbers/plate
TA98	DMSO	-	26.7	2.5	-	-	-	24, 29, 27
	Test item	5	26.7	6.4	1.0	-0.04	NS	34, 23, 23
		16	17.7	3.2	0.7	-1.78	NS	20, 19, 14
		50	20.3	4.0	0.8	-1.22	NS	25, 18, 18
		160	27.0	10.1	1.0	-0.04	NS	25, 18, 38
		500	12.7	3.2	0.5	-2.98	NS	9, 14, 15
		1600	8.0	2.0	0.3	-4.31	NS	8S, 10S, 6S
		5000	3.0	2.6	0.1	-6.88	NS	0SP, 4SP, 5SP
	B[a]P	10	296.0	81.7	11.1	-	-	324, 360, 204
TA100	DMSO	-	111.0	21.8	-	-	-	136, 96, 101
	Test item	5	113.0	11.1	1.0	0.21	NS	103, 125, 111
		16	118.0	18.4	1.1	0.59	NS	126, 97, 131
		50	102.0	19.3	0.9	-0.76	NS	80, 116, 110
		160	110.3	8.4	1.0	-0.01	NS	120, 106, 105
		500	65.7	7.2	0.6	-4.20	NS	74S, 61S, 62S
		1600	4.3	1.2	0.0	-14.73	NS	5V, 5V, 3V
		5000	-	-	-	-	-	-PT, -PT, -PT
	AAN	5	1045.7	47.5	9.4	-	-	1030, 1008, 1099
TA1535	DMSO	-	18.0	5.3	-	-	-	14, 16, 24
	Test item	5	12.3	3.2	0.7	-1.32	NS	16, 10, 11
		16	10.7	8.3	0.6	-2.04	NS	20, 4, 8
		50	12.7	6.8	0.7	-1.39	NS	5, 18, 15
		160	9.7	1.5	0.5	-2.03	NS	10, 11, 8
		500	13.7	2.5	0.8	-0.96	NS	16, 14, 11
		1600	5.0	1.0	0.3	-3.63	NS	6S, 5S, 4S
		5000	4.7	1.2	0.3	-3.77	NS	6SP, 4SP, 4MSP
	AAN	5	183.3	46.2	10.2	-	-	130, 212, 208
TA1537	DMSO	-	8.0	2.6	-	-	-	5, 10, 9
	Test item	5	10.7	0.6	1.3	1.02	NS	11, 11, 10
		16	9.0	2.6	1.1	0.39	NS	11, 6, 10
		50	9.0	4.6	1.1	0.30	NS	8, 5, 14
		160	8.0	2.0	1.0	0.03	NS	8, 10, 6
		500	4.7	0.6	0.6	-1.41	NS	4, 5, 5
		1600	2.0	1.7	0.3	-3.60	NS	0S, 3S, 3S
		5000	-	-	-	-	-	-PT, -PT, -PT
	AAN		245.7	45.7	30.7	-	-	196, 286, 255
TA102	DMSO	-	205.3	23.4	-	-	-	188, 196, 232
	Test item	5	222.3	22.5	1.1	0.85	NS	230, 240, 197
		16	218.3	14.6	1.1	0.67	NS	223, 202, 230
		50	245.7	1.5	1.2	1.98	NS	243, 244, 247
		160	228.3	44.2	1.1	1.09	NS	187,223, 275
		500	132.0	18.1	0.6	-4.15	NS	151S, 130S, 115S
		1600	-	-	-	-	-	-T, -T, -T
		5000	-	-	-	-	-	-PT, -PT, -PT
	AAN	20	1697.3	111.4	8.3	-	-	1686, 1589, 1817

P: precipitation of test item observed

S: slight thinning of background bacterial lawn

T: toxic, no revertant colonies

V: very thin background bacterial lawn

 

Table 3       Raw plate counts and calculated mutagenicity data, experiment 2 (- S9 mix)

 

Strain	Compound	Conc. level (µg/plate)	Mean revertants per plate	SD	Fold increase	Dunnett’s T-value	Statistic. sig.	Revertant numbers/plate
TA98	DMSO	-	22.0	7.2	-	-	-	16, 30, 20
	Test item	50	23.3	8.0	1.1	0.24	NS	24, 15, 31
		100	23.7	8.5	1.1	0.29	NS	24, 32, 15
		200	15.7	5.0	0.7	-1.34	NS	11, 15, 21
		400	16.0	3.5	0.7	-1.22	NS	14, 20, 14
		800	11.0	1.0	0.5	-2.46	NS	10S, 11S, 12S
		1600	2.7	2.1	0.1	-5.71	NS	2S, 1S, 5S
	2NF	5	323.7	60.7	14.7	-	-	310, 271, 390
TA100	DMSO	-	74.3	7.5	-	-	-	82, 74, 67
	Test item	50	82.7	5.1	1.1	1.10	NS	87, 77, 84
		100	84.3	5.0	1.1	1.31	NS	85, 89, 79
		200	76.7	10.5	1.0	0.30	NS	87, 77, 66
		400	43.7	4.0	0.6	-4.66	NS	46S, 39S, 46S
		800	13.7	7.2	0.2	-11.58	NS	22S, 9S, 10S
		1600	-	-	-	-	-	-T, -T, -T
	NaN3	2	415.0	1.7	5.6	-	-	414, 414, 417
TA1535	DMSO	-	15.3	3.5	-	-	-	19, 15, 12
	Test item	50	12.0	3.6	0.8	-1.34	NS	16, 9, 11
		100	12.7	2.1	0.8	-1.02	NS	15, 11, 12
		200	11.0	0.0	0.7	-1.70	NS	11, 11, 11
		400	15.0	4.0	1.0	-0.14	NS	15, 19, 11
		800	7.3	1.5	0.5	-3.51	NS	9S, 7S, 6S
		1600	2.0	1.7	0.1	-7.50	NS	1MSB, 1S, 4S
	NaN3	2	428.3	25.0	27.9	-	-	434, 450, 401
TA1537	DMSO	-	14.3	4.9	-	-	-	20, 11, 12
	Test item	50	10.7	5.0	0.7	-0.76	NS	16, 10, 6
		100	12.0	8.5	0.8	-0.62	NS	21, 4, 11
		200	11.0	5.0	0.8	-0.69	NS	6, 16, 11
		400	5.7	3.5	0.4	-2.02	NS	9S, 2S, 6S
		800	-	-	-	-	-	-T, -T, -T
		1600	-	-	-	-	-	-T, -T, -T
	AAC	50	86.7	32.1	6.0	-	-	100, 50, 110
TA102	DMSO	-	242.0	26.5	-	-	-	271, 236, 219
	Test item	50	229.3	28.1	0.9	-0.25	NS	256, 232, 200
		100	142.7	87.2	0.6	-2.51	NS	42, 195, 191
		200	164.7	10.8	0.7	-1.65	NS	160S, 157S, 177S
		400	105.7	9.1	0.4	-3.20	NS	102S, 99S, 116S
		800	-	-	-	-	-	-T, -T, -T
		1600	-	-	-	-	-	-T, -T, -T
	MMC	0.2	594.7	30.7	2.5	-	-	579, 575, 630

P: precipitation of test item observed

S: slight thinning of background bacterial lawn

T: toxic, no revertant colonies

V: very thin background bacterial lawn

 

Table 4       Raw plate counts and calculated mutagenicity data, experiment 2 (+ S9 mix)

 

Strain	Compound	Conc. level (µg/plate)	Mean revertants per plate	SD	Fold increase	Dunnett’s T-value	Statistic. sig.	Revertant numbers/plate
TA98	DMSO	-	27.7	6.7	-	-	-	20, 32, 31
	Test item	50	23.3	5.5	0.8	-0.92	NS	27, 26, 17
		100	35.7	3.5	1.3	1.59	NS	32, 36, 39
		200	30.0	10.5	1.1	0.41	NS	29, 20, 41
		400	27.3	2.9	1.0	-0.02	NS	29S, 24S, 29S
		800	14.0	3.0	0.5	-3.25	NS	14S, 11S, 17S
		1600	12.3	3.2	0.4	-3.76	NS	10S, 16S, 11S
	B[a]P	10	304.3	44.3	11.0	-	-	354, 290, 269
TA100	DMSO	-	115.7	21.8	-	-	-	92, 120, 135
	Test item	50	113.7	4.2	1.0	-0.09	NS	117, 109, 115
		100	104.0	5.3	0.9	-0.80	NS	102, 100, 110
		200	107.0	24.9	0.9	-0.65	NS	134, 102, 85
		400	53.0	10.4	0.5	-5.30	NS	59S, 41S, 59S
		800	24.0	8.7	0.2	-8.97	NS	34S, 19S, 19S
		1600	-	-	-	-	-	-T, -T, -T
	AAN	5	688.0	77.4	5.9	-	-	774, 666, 624
TA1535	DMSO	-	15.3	0.6	-	-	-	15, 16, 15
	Test item	50	20.0	5.2	1.3	1.83	NS	17, 17, 26
		100	17.0	3.0	1.1	0.68	NS	20, 14, 17
		200	14.7	3.2	1.0	-0.35	NS	16, 17, 11
		400	15.0	1.0	1.0	-0.15	NS	14S, 15S, 16S
		800	8.3	2.3	0.5	-3.59	NS	7S, 7S, 11S
		1600	7.3	1.5	0.5	-4.18	NS	6V, 9V, 7V
	AAN	5	167.7	11.0	10.9	-	-	179, 157, 167
TA1537	DMSO	-	11.7	2.1	-	-	-	11, 10, 14
	Test item	50	11.7	4.5	1.0	-0.09	NS	12, 7, 16
		100	14.0	5.2	1.2	0.70	NS	11, 11, 20
		200	11.3	2.5	1.0	-0.13	NS	11, 14, 9
		400	12.7	3.1	1.1	0.32	NS	12S, 16S, 10S
		800	-	-	-	-	-	-T, -T, -T
		1600	-	-	-	-	-	-T, -T, -T
	AAN	5	219.3	82.9	18.8	-	-	169, 174, 315
TA102	DMSO	-	277.3	19.1	-	-	-	257, 280, 295
	Test item	50	262.0	2.0	0.9	-0.97	NS	262, 264, 260
		100	266.7	10.7	1.0	-0.67	NS	260, 279, 261
		200	253.0	25.6	0.9	-1.58	NS	226, 277, 256
		400	199.3	19.4	0.7	-5.33	NS	209S, 212S, 177S
		800	107.0	14.1	0.4	-13.25	NS	105S, 122S, 94S
		1600	-	-	-	-	-	-T, -T, -T
	AAN	20	1826.3	118.6	6.6	-	-	1711, 1820, 1948

P: precipitation of test item observed

S: slight thinning of background bacterial lawn

T: toxic, no revertant colonies

V: very thin background bacterial lawn

 

 

Applicant's summary and conclusion

Conclusions:

The test item was considered to be non-mutagenic under the conditions of the test.

Executive summary:

OECD 471 (2017) - In a reverse gene mutation assay in bacteria, strains TA98, TA100, TA1535, TA1537 and TA102 of S. typhimuriumwere exposed to Amines, C12-14-branched alkyl dodecylbenzenesulfonates (1:1) in DMSO at concentrations of 5, 16, 50, 160, 500, 1600 and 5000 µg/plate a screening experiment followed by concentrations of 50, 100, 200, 400, 800 and 1600 µg/plate in a secondary, pre-incubation, experiment. Both tests were conducted in the presence and absence of S9 mix.

 

The test item was tested up to cytotoxic and precipitating concentrations. The positive controls induced the appropriate responses in the corresponding strains.

 

There was no evidence of induced mutant colonies over background in the test item treated colonies.

 

This study is classified as acceptable. This study satisfies the requirement for Test Guideline OECD 471 for in vitro mutagenicity (bacterial reverse gene mutation) data.