Phosphorous acid, tri-C12-14-alkyl esters

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:

April - May 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Report date:

2012

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:

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

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Phosphorous acid, tri-C12-14-alkyl esters

EC Number:

297-701-9

EC Name:

Phosphorous acid, tri-C12-14-alkyl esters

Cas Number:

93686-48-7

Molecular formula:

C36H75O3P C38H7903P C40H83O3P C42H87O3P

IUPAC Name:

Phosphorous acid, tri-C12-14-alkyl esters

Test material form:

other: liquid

Details on test material:

Batch no 80627X065A

Method

Target gene:

Genes involved in the functional capability of the bacteria to synthesize histidine

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA 1535, TA 97a, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

5000 / 1500 / 500 / 150 / 50 µg/plate for the first experiment (plate incorporation method) and 5002 / 2501 / 1251 / 626 / 313 µg/plate (for the pre incubation method)

Vehicle / solvent:

tetrahydrofurane (THF) was chosen as vehicle, because the test item was completely soluble, and addition of 5% of this solvent doesn’t have any effects on the viability of the bacteria or the number of spontaneous revertants

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

sodium azide

benzo(a)pyrene

other: 2-Amino-anthracene and 4-Nitro-1,2-phenylene diamine

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation) for the first experiment, then preincubation method for the second one.

DURATION:
- Preincubation period: incubation over night at 37 °C before the cultures were used in the experiment
- Exposure duration: 48h

SELECTION AGENT (mutation assays): histidine

NUMBER OF REPLICATIONS: four

DETERMINATION OF CYTOTOXICITY:
- Method: background lawn visible or not and the number of revertant colonies reduced or not

Evaluation criteria:

A test item is considered to have mutagenic potential, if a significant, reproducible in-crease of revertant colonies per plate (increase factor  2) in at least one strain can be observed. A concentration-related increase over the range tested can also be taken as a sign of mutagenic activity

Results and discussion

Test results

Key result

Species / strain:

S. typhimurium, other: TA 1535, TA 97a, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

Interpretation of results:
Negative with and without metabolic activation


The test item Phosphorous acid, tri-C12-14-alkyl esters was stated as not mutagenic under the test conditions.

Executive summary:

Phosphorous acid, tri-C12-14-alkyl esters was tested in vitro in an Ames test with Salmonella typhimurium.

Five concentrations of the test item, dissolved in tetrahydrofurane (THF) (ranging from 5000 to 50 µg/plate) were used in a first plate incorporation method, then five concentrations (ranging from 5001 to 313 µg/plate) were tested in a second experiment via pre incubation method.

Five genetically manipulated strains of Salmonella typhimurium (TA 97a, TA 98, TA 100, TA 102 and TA 1535) were exposed to the test item both in the presence and in the absence of a metabolic activation system (S9) for 48 hours.

None of the concentrations in the two experiments caused a significant increase in the number of revertant colonies in the tested strains.

No signs of toxicity towards the bacteria could be observed.

The sterility control and the determination of the titre didn’t show any inconsistencies. The determined values for the spontaneous revertants of the negative controls were in the normal range. All positive controls showed mutagenic effects with and without metabolic activation.

The study was then considered as valid and the test item didn’t show mutagenic effects towards Salmonella typhimurium, strains TA 97a, TA 98, TA 100, TA 102 and TA 1535.

Therefore, no concentration-effect relationship could be determined.

The test item Phosphorous acid, tri-C12-14-alkyl esters is considered as “not mutagenic under the conditions of the test”.