Registration Dossier

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:
September 22, 1999 - October 7, 1999
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Test was conducted similar to OECD Test Guideline No. 471, under early GLP Standards and QA, no report summary, no second experiment using pre-incubation method was performed. Acceptable basic data.
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1999
Report Date:
1999

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Dihydro Linalool
- Physical state: liquid

Method

Target gene:
His-gene: Amino acid histidine
Species / strain
Species / strain:
other: S. typhimurium TA 1535, TA97, TA 98, TA 100, and TA 1028
Details on mammalian cell lines (if applicable):
Not applicable
Additional strain characteristics:
other: all strains: rfa, uvrB; TA 97,TA 98, TA 100, TA 102: pKM101
Metabolic activation:
with and without
Metabolic activation system:
Rat liver S9 induced by phenobarbital/5,6 benzoflavone
Test concentrations with justification for top dose:
Plate incorporation assay:0, 10, 31.6, 100, 316, 1000 µg/plate
Vehicle:
- Vehicle(s)/solvent(s) used: DMSO
Controls
Negative controls:
no
Remarks:
absence of untreated test strain cultures
Solvent controls:
yes
Remarks:
DMSO
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: TA1535 and TA100 (- S9): Sodium Azide; TA98 and TA 1538 (- S9): 2-Nitrofluorene; TA 1537 (-S9): 9-Aminoacridine; all strains (+ S9): 2-Aminoanthracene
Details on test system and conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Selection time (if incubation with a selection agent): approx. 2 days

SELECTION AGENT (mutation assays): overlay agar containing Histidine

NUMBER OF REPLICATIONS: 3 plates per concentration

NUMBER OF CELLS EVALUATED: not applicable

DETERMINATION OF CYTOTOXICITY
- Method: In a preliminary toxicity assay determination of reduction in the revertant colony number, appearance of microcolonies, and/or observation of thinning or absence of the background lawn. Each test substance dose (14 doses of 0.02 to 150 ul/plate), as well as the appropriate solvent control, was evaluated in strain TA100 in the standard plate incorporation assay. In TA 100 the test material was completely toxic to revertant colonies at 9.38 ul/plate.
Evaluation criteria:
The criteria used to determine positive effects were based primarily on a historical data base. Most data sets were evaluated using the following criteria:
If the solvent control value is within the normal range, a test material producing a positive response equal to three times the solvent control value is considered mutagenic for strains TA 1535, and equal to twice the solvent control value for strains TA 98 and TA 100.
The following ranges of revertants for solvent controls are generally considered acceptable:
TA 1535: 8-30; TA 98: 20-75; TA 100: 80-250.
Statistics:
Mean values and standard deviation (SD).

Results and discussion

Test results
Species / strain:
other: S. typhimurium TA 1535, TA, 97, TA 98, TA 100, TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
yes
Remarks:
toxic to revertant colonies at 10 ul/plate
Vehicle controls valid:
yes
Negative controls valid:
not applicable
Positive controls valid:
yes
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Additional information on results:
Not relevant

Any other information on results incl. tables

Not relevant

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative with metabolic activation
negative without metabolic activation

The test material, did not exhibit genetic activity in any of the assays conducted for this evaluation and was considered not mutagenic under these test conditions. It was concluded that linalool does not need to be classified as mutagenic according to the criteria outlined in Annex I of 1272/2008/EC and Annex VI of 67/548/EEC.
Executive summary:

The test compound was examined for mutagenic activity in a series of in vitro microbial plate incorporation assays employing five Salmonella typhimurium strains (TA1535, TA97, TA98, TA100, TA 102). The compound was tested directly and in presence of liver microsomal enzyme preparations from phenobarbital/5,6 benzophenone-induced rats. A negative control consisting of the solvent (DMSO) used for preparing the stock solution and subsequent dilutions of the test material, and specific positive compounds were assayed concurrently with the test material.

The results of the test conducted on Dihydrolinalool in the absence and presence of a rat liver metabolic activation system were negative. The test material, did not exhibit genetic activity in any of the assays conducted for this evaluation and was considered not mutagenic under these test conditions. It was concluded that Dihydrolinalool does not need to be classified as mutagenic according to the criteria outlined in Annex I of 1272/2008/EC and Annex VI of 67/548/EEC.