Nonanoic acid, mixed esters with heptanoic acid, pentaerythritol and valeric acid

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:

18 April - 20 June 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

21 July 1997

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Groupe Interministeriel des Produits Chimiques, France

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Monopentaerythritol tetraesters and dipentaerythritol hexaesters of valeric, heptanoic and nonanoic acids

Molecular formula:

not applicable, substance is UVCB

IUPAC Name:

Monopentaerythritol tetraesters and dipentaerythritol hexaesters of valeric, heptanoic and nonanoic acids

Method

Target gene:

"his operon" (for S. typhimurium strains) and "trp operon" (for E.coli strains)

Species / strain

Species / strain 1

Species / strain / cell type:

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

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A pKM 101

Metabolic activation:

with and without

Metabolic activation system:

Cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from livers treated with Aroclor 1254

Test concentrations with justification for top dose:

Preliminary toxicity study: 50, 150, 500, 1500 and 5000 µg/plate with and without metabolic activation
First experiment (direct incorporation): 50, 150, 500, 1500 and 5000 µg/plate with and without metabolic activation
Second experiment (pre-incubation) experiment: 50, 150, 500, 1500 and 5000 µg/plate with and without metabolic activation

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: ethanol

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

untreated control plates

Negative solvent / vehicle controls:

yes

Remarks:

ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Controls 2

Untreated negative controls:

yes

Remarks:

untreated control plates

Negative solvent / vehicle controls:

yes

Remarks:

ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Controls 3

Untreated negative controls:

yes

Remarks:

untreated control plates

Negative solvent / vehicle controls:

yes

Remarks:

ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Controls 4

Untreated negative controls:

yes

Remarks:

untreated control plates

Negative solvent / vehicle controls:

yes

Remarks:

ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: cis-platinum (II) diammine dichloride

Controls 5

Untreated negative controls:

yes

Remarks:

untreated control plates

Negative solvent / vehicle controls:

yes

Remarks:

ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-anthramine

Controls 6

Untreated negative controls:

yes

Remarks:

untreated control plates

Negative solvent / vehicle controls:

yes

Remarks:

ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: dimethylbenzanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION:
- First experiment: in agar (plate incorporation)
- Second experiment: preincubation

DURATION
- Exposure duration: 48 - 72 h

NUMBER OF REPLICATIONS: triplicates each in two independent experiments

DETERMINATION OF CYTOTOXICITY
- Method: inspection of the bacterial background lawn


OTHER: a preliminary toxicity test was carried out in the strains TA100

Evaluation criteria:

The result of the test is considered positive if a concentration dependent relationship is obtained in one or several of the 5 strains, with and/or without metabolic activation. A mutagenic effect is taken into account for a given test concentration if the number of revertant colonies is at least two-fold that of spontaneous revertant colonies for S. typhimurium TA 98, TA 100 and E. coli WP2 uvr A pKM 101 and three-fold for S. typhimurium TA 1535 and TA 1537. All results must be confirmed in an independent experiment.

Statistics:

Mean values and standard deviations were calculated.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 5

Species / strain:

E. coli WP2 uvr A pKM 101

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: no precipitate was observed at at any dose level tested

RANGE-FINDING/SCREENING STUDIES: The dose range for the main test was determined in a preliminary toxicity assay where the test material was tested at the following doses: 50, 150, 500, 1500 and 5000 µg/plate with and without metabolic activation. The test material was non-toxic to the strains of bacteria used (TA 100) at any dose level.

COMPARISON WITH HISTORICAL CONTROL DATA: All tester strain cultures exhibit a characteristic number of spontaneous revertants per plate in the vehicle and untreated controls. Furthermore positive control values were at least two times the respective vehicle control value for each strain. The historical control ranges are presented in the table below.

Any other information on results incl. tables

Due to the complexity and the amount of information, all relevant tables summarising results and historical control data are attached as pdf document (Tables_OECD 471.pdf).

Applicant's summary and conclusion

Conclusions:

Interpretation of results: negative