Dodecan-1-ol, ethoxylated

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:

data from handbook or collection of data

Justification for type of information:

data from handbook or collection of data

Data source

Reference

Reference Type:

publication

Title:

SALMONELLA MUTAGENICITY TESTS

Author:

ZEIGER,E et al.

Year:

1987

Bibliographic source:

ENVIRON. MOL. MUTAGEN.

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other:

Principles of method if other than guideline:

Preincubation Assay
The preincubation assay was performed . The test chemical, Salmonella culture, and S-9 mix or buffer 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 that contained Vogel-Bonner medium [Vogel and Bonner, 19561. The histidine-revertant (his') colonies arising on these plates were counted following 2 days incubation at 37°C. The plates were hand-counted when a precipitate was present; otherwise automatic colony counters were used.
All chemicals were tested initially in a toxicity assay to determine the appropriate dose range. The toxicity assay was performed by using TA100 or the system.
Toxic concentrations were those at which a decrease in the number of hisf colonies was seen or at which there was a clearing in the density of the background lawn.At least five doses of the chemical were tested in triplicate. Experiments were repeated at least 1 wk following the initial trial. Each chemical was tested initially at half-log doses up to a dose that elicited toxicity; subsequent trials occasionally used narrower dose increments. Chemicals that were not toxic were tested to a maximum dose of 10 mglplate. Chemicals that were poorly soluble were tested up to a dose defined by their solubility. A maximum of 0.05 ml solvent was added to each plate.Concurrent solvent and positive controls were run with each trial. The positive controls in the absence of metabolic activation were sodium azide (TA1535 and TA loo), 9-aminoacridine (TA97 and TA 1S37), and 4-nitro-o-phenylenediamine (TA98). The positive control for metabolic activation was 2-aminoanthracene for all strains. Although there were no specific response ranges established for the solvent and positive controls, each laboratory rejected experiments in which the positive control chemical did not produce a mutagenic response or in which the solvent controlvalues were higher (or lower in the case of TAlOO and TA97) than their expected values.During the initial stages of the testing program, chemicals were tested with 10% S-9. Other levels of S-9 were used when an equivocal result was obtained with 10%. The protocol evolved to one that used 30% S-9 when a negative response was obtained with 10% S-9.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Dodecan-1-ol, ethoxylated

EC Number:

500-002-6

EC Name:

Dodecan-1-ol, ethoxylated

Cas Number:

9002-92-0

Molecular formula:

C58H118024

IUPAC Name:

Dodecan-1-ol, ethoxylated

Test material form:

liquid

Details on test material:

Name: Dodecyl alcohol, ethoxylated
CAS No.: 9002-92-0
Molecular Formula: (C2-H4-O)mult-C12-H26-O
Molecular Weight: 230.389 g/mol

Method

Target gene:

histidine-manufacturing gene.

Species / strain

Species / strain / cell type:

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

Additional strain / cell type characteristics:

other: All the bacterial strains used in the Ames test carry a defective (mutant) gene that prevents them from synthesizing the essential amino acid histidine from the ingredients in standard bacterial culture medium.

Metabolic activation:

with and without

Metabolic activation system:

10% HLI =induced male Syrian hamster liver S9 ; 10% RLI = induced male Sprague Dawley rat liver S9

Test concentrations with justification for top dose:

0,1,3,10,33,100,333 and 1000 µg/Plate

Vehicle / solvent:

Vehicle Control : Dimethyl Sulfoxide

Controls

Controls 1

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

water

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

Positive control for TA 100 and TA 1535 tested in the absence of S9 Migrated to IUCLID6: without S9

Controls 2

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

Water

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

Positive control for TA 98 tested in the absence of S9

Controls 3

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

Water

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

Positive control for TA 1537 tested in the absence of S9

Controls 4

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

Water

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene (or occasionally, sterigmatocystin)

Remarks:

For all strains(TA 100,TA 1535, TA1537, TA98) tested with S9

Details on test system and experimental conditions:

All chemicals were tested in Salmonella typhimurium strains TA98, TA100,TA1535, and TA1537 and/or TA97. The majority of chemicals were tested in TA1537,and a few were tested in TA97. The testing in both strains is the result of an evolution of the protocol described by Haworth et a1 [1983]. In this original protocol, TA1537 was used. In a later protocol, TA97 replaced TA1537, but the option to retest a chemical in TA1537 was retained for chemicals that produced a positive or questionable response in TA97 and negative responses in the other strains.All strains were obtained from Dr. Bruce Ames and were stored as recommended [Maron and Ames, 19831. Prior to their use for mutagenicity assays, all cultures were grown overnight with shaking at 37°C in Oxoid broth, and their phenotypes were analyzed.

Evaluation criteria:

The histidine-revertant (his') colonies were observed for mutagenicity.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

with 10% HLI and 10% RLI

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

with 10% HLI and 10% RLI

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

with 10% HLI and 10% RLI

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

with 10% HLI and 10% RLI

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

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

TA 100

Dose	No Activation (Negative)		No Activation (Weakly Positive)		10% RLI (Negative)		10% RLI (Negative)		10% HLI (Negative)		10% HLI (Negative)
Protocol	Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation
ug/Plate	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM
0	99	2	98	3.8	137	1.7	94	5.5	143	5.5	111	7
1			119	7
3	95	3.2	132	3.8			115	6.5			122	8.7
10	98	6	137	5	164	7.4	107	3.7	139	9.7	123	11.8
33	97	8.6	120	8.8	151	8.2	127	1.8	135	4.6	120	5.2
100	61	5.3	105	4.6	157	3	111	8.1	146	7.7	116	11
333	0	0			139	8	60	6.2	136	19.7	85	12.5
1000					23	4.4			11	2.2
Positive Control	322	6	405	7.1	885	103.7	419	26.3	1065	143.4	1204	108.6

TA 1535

Dose	No Activation (Negative)		No Activation (Negative)		10% RLI (Negative)		10% RLI (Negative)		10% HLI (Negative)		10% HLI (Negative)
Protocol	Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation
ug/Plate	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM
0	25	3.8	20	.9	11	1.5	7	.9	6	1.3	7	2.3
1	29	1.5
3	29	1.5	26	4.2			7	.6			10	1.5
10	29	4	33	2.9	11	1.5	10	1.5	10	1.5	5	2
33	23	4.3	27	4.9	13	.7	9	2.3	9	3.5	11	1.7
100	32	3.2	19	4.8	9	2.7	7	1.2	13	.7	12	1.8
333			14	.9	7	.9	7	.7	12	1.7	7	2
1000					8	.9			6	2
Positive Control	452	12.8	352	41.8	162	11.8	183	11.5	437	22.8	502	41

TA 1537

Dose	No Activation (Negative)		No Activation (Negative)		10% RLI (Negative)		10% RLI (Negative)		10% HLI (Negative)		10% HLI (Negative)
Protocol	Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation
ug/Plate	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM
0	3	1.3	5	1	9	2.2	4	.7	7	1.9	4	.9
1	7	2.5
3	7	.9	3	.6			8	2.1			10	2
10	7	1.3	5	2.3	6	.9	8	2.4	8	2.3	6	.3
33	6	1.9	3	.6	8	.6	9	1.5	7	2.8	8	.7
100	9	1.3	4	.7	10	1.2	6	.9	7	.3	8	1.2
333			2	.9	4	1.2	6	.6	9	2.1	6	.3
1000					4	.3			4	.6
Positive Control	149	27.8	110	9.7	116	23.4	122	17.1	409	14.9	246	27.1

TA 98

Dose	No Activation (Negative)		No Activation (Negative)		10% RLI (Negative)		10% RLI (Negative)		10% HLI (Negative)		10% HLI (Negative)
Protocol	Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation
ug/Plate	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM
0	16	.7	16	2.1	24	3.9	31	3.6	31	0	30	3.2
1	14	2.6
3	16	1.2	13	.6			32	4.1			32	3.5
10	20	2.3	15	.7	25	5.7	29	1.5	36	2	35	4.3
33	22	3.9	11	2.3	27	3.8	26	2.3	32	1.3	35	1.8
100	17	4.7	13	2.5	29	2	31	3.5	29	.6	35	2.2
333			8	2.5	24	6.3	34	2.2	25	3	39	.3
1000					29	2.5			28	1.8
Positive Control	830	6	596	22.2	414	16.7	409	8.5	827	37.4	1072	104.4

RLI = induced male Sprague Dawley rat liver S9
HLI = induced male Syrian hamster liver S9

s = Slight Toxicity; p = Precipitate; x = Slight Toxicity and Precipitate; T = Toxic; c = Contamination

Applicant's summary and conclusion

Conclusions:

In an Ames test , test chemical dissolved in dimethyl sulfoxide from doses 0 - 1000 micrograms per plate was not mutagenic in Salmonella typhimurium strain TA 100, TA1535, TA1537 and TA98 with and without addition of S9 liver fractions from Aroclor induced hamsters and rats.

Executive summary:

Genetic toxicity in vitro study was assessed for test chemical. For this purpose AMES test was performed .The test material was exposed to Salmonella typhimurium TA100, TA1535, TA98 and TA1537in the presence and absence of metabolic activation S9. The concentration of test material used in the presence and absence of metabolic activation were 0 ,1,3,10,33,100,333and 1000 µg/plate. No mutagenic effects were observed in all strains, in the presence and absence of metabolic activation. Therefore test chemical was considered to be non-mutagenic in Salmonella typhimurium TA100, TA1535, TA98 and TA1537by AMES test. Hence the substance cannot be classified as gene mutant in vitro.