esterification product of 2-hydroxypropane-1,3-diyl-bis(2-methylprop-2-enoate) and 3-hydroxypropane-1,2-diyl-bis(2-methylprop-2-enoate) and diphosphorus pentoxide

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:

November to December 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted under GLP conditions.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Report date:

2001

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Glypho

IUPAC Name:

Glypho

Details on test material:

- Name of test material (as cited in study report:Glypho
- Substance type:Yellowish liquid
- Physical state:Liquid
- Lot/batch no.:AH2/170
- Expiration date of the lot/batch: February 01, 2002
- Stability under test conditions:Stable 48 hours in water and polyethylene glycol
- Storage condition of test material: at 4 degrees C

Method

Target gene:

histidine gene locus (Salmonella), tryphtophan biosynthesis (E. coli)

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Additional strain / cell type characteristics:

other: All strains have decreased DNA excision repair system, AMP resistance, altered permeability

Metabolic activation:

with and without

Metabolic activation system:

S9 liver microsomal fraction

Test concentrations with justification for top dose:

33, 100, 333, 1000, 2500, and 5000 micrograms/plate. The test article was tested at 5000 micrograms/plate as a maximal concentration in both main experiments.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: Solubility properties and its relative non-toxicity to the bacteria.

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: Sodium azide, 4-nitro-o-phenylene-diamine, methyl methane sulfonate (all 3 without metabolic activation), and 2-aminoanthracene (with metabolic activation)

Details on test system and experimental conditions:

METHOD OF APPLICATION:preincubation and in agar (plate incorporation)
DURATION
- Preincubation period:60 minutes
- Exposure duration:48 hours
- Expression time (cells in growth medium):48 hours
- Fixation time (start of exposure up to fixation or harvest of cells):49 hours
NUMBER OF CELLS EVALUATED: All colonies counted using the AUTOCOUNT (Artek Systems Corporation)
DETERMINATION OF CYTOTOXICITY
- Method:relative total growth
OTHER: Media: Histidine-free (Salmonella), tryptophan-free (E. coli)

Evaluation criteria:

A test item is considered as a mutagen if a biologically relevant increase in the number of revertants exceeding the threshold of twice (strains TA 98, TA 100, and WP2 uvrA) or thrice (strains TA 1535 and TA 1537) the colony count of the corresponding solvent control is observed.

Results and discussion

Test results

Species / strain:

other: TA 98, TA 100, TA 1535, TA1537 and WP2 uvrA

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: moderate toxic effects, evident as reduction of number of revertant, in TA 1537 in first experiment at high concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: pH of test article adjusted with NaOH
RANGE-FINDING/SCREENING STUDIES: Yes; concentration range 3-5000 micrograms/plate
COMPARISON WITH HISTORICAL CONTROL DATA: Yes

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

The test article was not mutagenic in the Bacterial Reverse Mutation Assay in the presence or absence of the S9-mix.

Executive summary:

The mutagenic potential of the test article was evaluated in the Bacterial Reverse Mutation Assay with S. typhimurium strains TA98, TA100, TA1535 and TA1537 and E. coli strain WP2 uvrA in the presence and absence of a metabolic activation system (S9-mix: phenobarbital and B-naphthoflavone-induced rat liver). This study was performed in compliance with OECD GLP (1997) and the Chemicals Act of Germany (1994). The study design was based on OECD 471 (1997) and Commission Directive 2000/32/EC L1362000, An 4D (2000).The test article was diluted in DMSO prior to administration to the cells. The mutagenic assay was performed in each strain at 33, 100, 333, 1000, 2500 or 5000 ug/plate in the presence or absence of S9-mix. Strain specific positive controls were performed in parallel. All treatments were tested in triplicate. Moderate toxic effects, evident as a reduction in number of revertants, occurred in strain TA1537 at 2500 and 5000 ug/plate with metabolic activation. No increase in revertant colonies was observed in either the presence or absence of S9-mix for any strain. The control results for strain TA1535 (positive control without S9-mix), WP2 uvrA (positive control with S9-mix), and TA1537 (solvent and positive control with S9-mix) were not quite within the historical control range. These effects are based on biological fluctuations and have no impact on the outcome of the study. Strain specific positive controls showed a distinct increase of induced revertant colonies. The study results were considered valid. Under the conditions of this study the test article was not mutagenic in the Bacterial Reverse Mutation Assay in the presence or absence of S9-mix.