2-({[4-({bis[4-({[2-(prop-2-enoyloxy)ethoxy]carbonyl}amino)phenoxy](sulfanylidene)-$l^{5}-phosphanyl}oxy)phenyl]carbamoyl}oxy)ethyl prop-2-enoate

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:

2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

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

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:

2-({[4-({bis[4-({[2-(prop-2-enoyloxy)ethoxy]carbonyl}amino)phenoxy](sulfanylidene)-λ⁵-phosphanyl}oxy)phenyl]carbamoyl}oxy)ethyl prop-2-enoate

EC Number:

918-919-9

Cas Number:

1072454-85-3

Molecular formula:

C36H36N3O15PS

IUPAC Name:

2-({[4-({bis[4-({[2-(prop-2-enoyloxy)ethoxy]carbonyl}amino)phenoxy](sulfanylidene)-λ⁵-phosphanyl}oxy)phenyl]carbamoyl}oxy)ethyl prop-2-enoate

additive 1

Reference substance name:

Ethyl acetate

EC Number:

205-500-4

EC Name:

Ethyl acetate

Cas Number:

141-78-6

Molecular formula:

C4H8O2

IUPAC Name:

ethyl ethanoate

Test material form:

liquid

Method

Target gene:

mutant histidine gene

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:

S9 mix was made from the livers of Aroclor 1254-induced male Sprague Dawley rats. The S9 mix comprised 10 % S9 fraction.

Test concentrations with justification for top dose:

The test item is a 40 % solution in ethyl acetate. This content was taken into consideration for the calculation of the doses tested:
plate incorporation assay:
0, 10, 25, 50, 160, 500, 1600, 5000 µg/plate with and without S9 mix
pre-incubation assay:
0, 10, 25, 50, 160, 500, 1600, 5000 µg/tube with and without S9 mix
The test was performed up to and including the limit dose of 5000 µg/plate and tube. No bacteriotoxicity but substance precipitation occurred at this dose.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: ethyl acetate
- Justification for choice of solvent/vehicle: test item is already a solution in ethyl acetate

Controls

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

2-nitrofluorene

sodium azide

cumene hydroperoxide

mitomycin C

other: 4-nitro-o-phenylenediamine for TA 1537; 1-Anthracene-2-amine for TA 102 and S9-mix activity

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation); pre-incubation

DURATION
- Pre-incubation period: 20 min
- Exposure duration: 48 hours (TA102) or 72 hours (TA1535, TA100, TA1537, TA98)

NUMBER OF REPLICATIONS: all plates were prepared in triplicate

DETERMINATION OF BACTERIOTOXICITY
- Method: gross appraisal of background growth

Evaluation criteria:

A reproducible and dose-related increase in mutant counts of at least one strain is considered to be a positive result. For TA 1535, TA 100, TA 1537 and TA 98 this increase should be about twice that of negative controls. For TA 102 an increase of about 100 mutants should be reached. Otherwise, the result is evaluated as negative. However, these guidelines may be overruled by good scientific judgment.
In case of questionable results, investigations should continue, possibly with modifications, until a final evaluation is possible.

Statistics:

no statistics perfomed; evaluation based on criteria mentioned above

Results and discussion

Test results

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Test item precipitation occurred at 5000 µg per plate/tube. No bacteriotoxic effects were seen up to and including 5000 µg/plate and tube. Bacteriotoxic effects were observed at 5000 µg/plate and tube.
No indication of mutagenic effects could be found for the test item at doses of up to and including 5000 µg per plate in any of the Salmonella typhimurium strains used, without and with metabolic activation, under the experimental conditions applied.

Remarks on result:

other: precipitation occured at the highest dose tested

Applicant's summary and conclusion

Executive summary:

The test item was evaluated in an Ames Test on Salmonella typhimurium strains TA 1535, TA 100, TA, 1537, TA 98, and TA 102, performed according to OECD TG 471. Doses of up to and including 5000 µg per plate did not produce bacteriotoxic effects. Substance precipitation occurred at 5000 µg per plate. The test material was considered to be non-mutagenic without and with S9 mix in the plate incorporation as well as in the pre-incubation modification of the Salmonella/microsome test. Positive controls increased the mutant counts to well over those of the negative controls, thus demonstrated the system´s sensitivity.