Anthracene oil

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:

08 Aug. - 06 Sep. 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Principles of method if other than guideline:

Two replicate experiments: plate incorporation assay and preincubation test

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Anthracene oil

EC Number:

292-602-7

EC Name:

Anthracene oil

Cas Number:

90640-80-5

Molecular formula:

Not applicabel

IUPAC Name:

Anthracene oil

Test material form:

other: depending on composition (UVCB) solid or viscous liquid

Details on test material:

anthracene oil; anthracene oil ≤ 50 ppm BaP, AOL (anthracene oil low)
- substance type: organic

Specific details on test material used for the study:

- Name of test material (as cited in study report): anthracene oil (< 50 ppm BaP)
- Lot/batch No. of test material: ATE No. 10443
- Stability under test conditions: no measured data; based on chemical structure assumed to be stable
- Storage condition of test material: room temperature, exclusion of light

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:

Microsomal fraction prepared from induced livers of male Wistar rats, induced with phenobarbital (80 mg/kg bw) and ß-naphthoflavone (100 mg/kg bw) orally (3x)

Test concentrations with justification for top dose:

1st experiment: 3.16, 10, 31.6, 100, 316, and 1000 µg/plate (TA 98, TA 100, -S9)
1.0, 3.16, 10, 31.6, 100, 316, and 1000 µg/plate (TA 1535, TA 1537, TA 102, - S9)
3.16, 10, 31.6, 100, 316, 1000 and 2500 µg/plate (+ S9)
2nd experiment: with variations based on results of 1st experiment (Report, p. 11, p. 19)
3rd experiment: 200, 400, 500, 600, 750, and 1000 (only with TA 100, +S9)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: compatible with survival of bacteria and S9 activity

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: see Report p. 15

Details on test system and experimental conditions:

METHOD OF APPLICATION:
1st experiment: in agar (plate incorporation)
2nd and 3rd experiment: pre-incubation variant

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth/colony formation

Evaluation criteria:

Considered as mutagenic
- if a clear and dose-related increase in the number of revertants occurs in at least one tester strain with or without metabolic activationand/or
- if a biologically relevant positive response for at least one of the dose groups occurs in at least one tester strain with or without metabolic activation.

An increase is considered relevant
- if in TA 100 and TA 102 mutation rate is at least twice as high as the rate of the solvent control;
- if in TA 98, TA 1535, and TA 1537 the mutation rate is at least 3 fold higher than that of the solvent control.

Statistics:

According to the OECD guidelines, the biological relevance is the criterion for the interpretation of the results: a statistical evaluation was not considered necessary under this premise (report p. 21).

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with

Genotoxicity:

positive

Remarks:

in 2nd and 3rd test: reproducible at cytotoxic concentrations (MF = 2.2 and 2.3)

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at ≥ 200 µg/pl. (+S9)

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

cytotoxic at higher concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium, other: TA 98, TA 1535, TA 1537, TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

reproducible in both tests

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

cytotoxic at higher concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

No mutagenic activity was observed in any test (plate incorporation and pre-incubation assay) at non-cytotoxic concentrations. In the pre-incubation assay, a mutagenic response was only observed in tester strain TA 100 with metabolic activation and at cytotoxic concentrations.

Experiment I (plate incorporation assay): No mutagenic activity was observed at all concentrations with and without metabolic activation.

Experiment II (pre-incubation assay): Relevant increase in the number of revertants in TA 100 but only at cytotoxic concentrations (500 µg/pl., cytotoxic, +S9) with MF = 2.0

Experiment III (pre-incubation assay): Relevant increases in revertants in TA 100 but only at cytotoxic concentrations (≥ 200 µg/plate, cytotoxic, +S9) with MF = 2.2 and 2.3.

Applicant's summary and conclusion

Conclusions:

From the five tester strains tested, only one (TA 100) showed a weak positive response with metabolic activation at already cytotoxic concentrations.