Decahydronaphthalene

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

Study period:

1989-09-25 to 1989-10-31

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study with acceptable restrictions: TA 102 or E.coli WP2 were not tested (not required by applied version of guideline).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1989

Report date:

1989

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

Cited as Directive 84/449/EEC, B.14

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Decahydronaphthalene

EC Number:

202-046-9

EC Name:

Decahydronaphthalene

Cas Number:

91-17-8

Molecular formula:

C10H18

IUPAC Name:

decahydronaphthalene

Details on test material:

Decahydronaphthalene of Hüls AG, purity 99 %, sample ID 734/881018

Method

Target gene:

mutated gene loci responsible for histidine auxotrophy

Species / strain

Species / strain / cell type:

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

Metabolic activation:

with and without

Metabolic activation system:

  Aroclor 1254 induced rat S9 liver, male Bor: WISW (SPF/Cpb) rats

Test concentrations with justification for top dose:

8; 40; 200; 1000; 5000 µg/plate

Vehicle / solvent:

solvent acetone (CAS No. 67-64-1)

Controls

Untreated negative controls:

yes

Remarks:

untreated 

Negative solvent / vehicle controls:

yes

Remarks:

solvent controls

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

sodium azide

other: Aminoacridine TA 1537

Details on test system and experimental conditions:

Ames test
SYSTEM OF TESTING
- Metabolic activation system:   Aroclor 1254 induced rat S9 liver, male Bor: WISW (SPF/Cpb) rats

ADMINISTRATION: 
- Dosing:    
main test: 8/40/200/1000/5000 µg/plate (+/- metabolic activation)   
pre-incubation test: 125/250/500/1000/2000 µg/plate (+/- metabolic  activation)
- Number of replicates: 3
- Application: 
solvent acetone (CAS No. 67-64-1)   main test 100 g/l, pre-incubation test 80 g/l
- Positive and negative control groups and treatment:    
positive, TA 98 and TA 1538: nitrofluorene   
positive, TA 100 and TA 1535: sodium azide   
positive, TA 1537: aminoacridine   
negative: untreated + solvent controls   
activity of metabolic system: aminoanthracene / TA 100
- Pre-incubation: 30 minutes at 30 °C   
incubation 96 hours at 37 °C

Evaluation criteria:

CRITERIA FOR EVALUATING RESULTS:    
mutagenic effects (i.e  ratio of revertant rates treated/control >= 2)  at <= 5000 µg/plate with generally positive dose-response relationship in  
any strain

Statistics:

no data

Results and discussion

Test results

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: > 5000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: other: Salmonella typhimurium TA 1535, TA 1537, TA 1538, TA 98, TA 100

Any other information on results incl. tables

GENOTOXIC EFFECTS: 
- With metabolic activation: None
- Without metabolic activation: Slight effects in TA 98 and TA 1537 in  the main test and slight effects in  TA 1537 in the pre-incubation test  were associated with very low revertant rates in solvent control and not  dose-related.

Thus they were not considered to be signs of mutagenicity:
  -------------------------------------------------------
  Revertant rates in pre-incubation test without metabolic activation:
  TA 1537, untreated: 8; solvent: 7; 125 µg: 7; 250 µg: 9; 500 µg: 7;  1000 µg: 15; 2000 µg: 15
  -------------------------------------------------------
  Revertant rates in main test without metabolic activation:
  TA 98, untreated: 11; solvent: 4; 8 µg: 4; 40 µg: 10; 200 µg: 4; 1000  µg: 5; 5000 µg: 15, precipitation
  TA 1537, untreated: 7; solvent: 4; 8 µg: 7; 40 µg: 4; 200 µg: 12; 1000  µg: 3; 5000 µg: 0, toxic and precipitation
  -------------------------------------------------------
  The positive controls were functional.
PRECIPITATION CONCENTRATION: 5000 µg/plate
CYTOTOXIC CONCENTRATION: 
  TA 98: None (+/- S9)
  TA 100: None (+/- S9)
  TA 1535: 5000 µg/plate (+/- S9)
  TA 1537: 5000 µg/plate (- S9)
  TA 1538: None (+/- S9)

Applicant's summary and conclusion

Conclusions:

Decahydronaphthalene was not genotoxic in both the presence or absence of metabolic activation in a microbial mutagenicity assay.

Executive summary:

In a reverse gene mutation assay in bacteria, strains TA98, TA100, TA1535, TA1537 and TA1538 of Salmonella typhimurium were exposed to decahydronaphthalene using acetone as a vehicle at concentrations of 8 - 5000 µg/plate, with and without S-9 activation in a preincubation assay according to EEC guidance Dir. 84/449/EEC, B.14.

The test article was tested at six dose levels along with appropriate vehicle and positive controls on the tester strains mentioned above in the presence and absence of S9-mix. All dose levels, vehicle and positive controls were plated in triplicate.

Cytotoxicity was noted at 1000 -5000 µg/plate.

No positive mutagenic responses were observed with any of the strains used, in the presence as well as in the absence of microsomal enzymes.These results were confirmed in an independent assay.