Alcohols, C12-18, ethoxylated

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

weight of evidence

Study period:

From 25 August 1996 to 30 September 1996

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study with acceptable restrictions.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

Neither E.coli WP2 strains nor S. typhimurium TA102 were used.

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

Neither E.coli WP2 strains nor S. typhimurium TA102 were used.

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Alcohols, C12-14, ethoxylated

EC Number:

500-213-3

EC Name:

Alcohols, C12-14, ethoxylated

Cas Number:

68439-50-9

IUPAC Name:

68439-50-9

Details on test material:

- Name of test material (as cited in study report): Lauryl polyglycol ether
- Ethoxylation degree: 2
- Analytical purity: No data
- Physical state: Clear colourless liquid
- Lot/batch No.: 962190
- Storage condition of test material: Ambient <25ºC, shielded from light

Method

Target gene:

His-operon

Species / strain

Species / strain 1

Species / strain / cell type:

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

Additional strain / cell type characteristics:

not applicable

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1538

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

S9 liver microsomal fraction

Test concentrations with justification for top dose:

without metabolic activation: 0, 1.5, 5, 15, 50, 150 and 500 µg/plate
with metabolic activation: 0, 5, 15, 50, 150, 500, 1500 and 5000 µg/plate

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: N-ethyl-N'-nitrosoguanidine (3-5 µg/plate), 9-aminoacridine (80 µg/plate), 4-Nitro-o-phenylenediamine (5 µg/plate), 4-nitroquinoline-N-oxide (0.2 µg/plate), 2-aminoanthracene (0.5-2 µg/plate)

Details on test system and experimental conditions:

METHOD OF APPLICATION:
A known aliquot (0.5 mL) of S9-mix and 2 mL of molten, trace histidine supplemented media were overlaid onto a sterile Vogel-Bonner Minimal agar plate in order to assess the sterility of the S9-mix.

Incubation and Assessment of Plates: all of the plates were incubated at 37 ºC for approximately 48 h and the frequency of revertant colonies assessed using a Domino colony counter.

NUMBER OF REPLICATIONS: Triplicate

DETERMINATION OF CYTOTOXICITY
- Method: The cytotoxicity was rated as follows:
S= sparse background lawn
V= very thin background lawn
T= toxic
NT= not tested at this dose level

Evaluation criteria:

The test item was considered positive if:
There is a dose related increase in the number of revertants or a biologically relevant increase for at least one test concentration.

Results and discussion

Test results

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

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

Number of Revertants (Number of Colonies per plate)
Base-pair Substitution Type		Frameshift Type
TA100	TA1535	TA1538	TA98	TA1537
147	33	29	29	9
124    (133)	32    (34)	21  (25)	40   (31)	7   (10)
129	36	24	33	13
			2225 (22)b19
Number of Revertants (Number of Colonies per plate)
Base-pair Substitution Type		Frameshift Type
TA100	TA1535	TA1538	TA98	TA1537
88	15	9	23	7
93    (88)	19    (16)	7   (9)	22   (22)	8   (9)
84	15	10	21	12

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

The test material was found to be non-mutagenic.

Executive summary:

Salmonella typhimurium strains TA1535, TA1537, TA1538, TA98 and TA100 were treated with the test material using the Ames plate incorporation method in triplicates, both with and without the addition of a rat liver homogenate metabolising system (10% liver S9 in standard co-factors). The dose range was determined in a preliminary toxicity assay and ranged between 1.5 and 5000 µg/plate in the first experiment. The vehicle (dimethyl sulphoxide) control plates produced counts of revertant colonies within the normal range. All the positive control chemicals used in the test induced marked increases in the frequency of revertant colonies, both with and without the metabolising system. The test material caused a visible reduction in the growth of the bacterial lawn to all of the strains of salmonella tested. The sensitivity of the bacterial strains to the toxicity of the test material varied between strains and between exposures with or without S9 -mix. The first incidence of toxicity was recorded at 150µg/plate. The test material was, therefore, tested up to its toxic limit. The test material was considered to be NON-MUTAGENIC under the conditions of this test.