Pentaerythritol

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:

7 September 1993 to 1 December 1994

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1994

Report date:

1994

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Pentaerythritol

EC Number:

204-104-9

EC Name:

Pentaerythritol

Cas Number:

115-77-5

Molecular formula:

C5H12O4

IUPAC Name:

2,2-bis(hydroxymethyl)propane-1,3-diol

Details on test material:

Pentaerythritol (abbreviated to PE in the report), molecular weight 136.2, supplied as a white powder from Japan Chemical Industry Association, manufactured by Mitsubishi Gas Chemical Company. Purity is 92.7% (Lot number: 50825; Containing impurities such as Dipentaerythritol (2.2%), Bis-Pentaerythritol (4.9%), and unknown impurities of (0.2%).

Stability tests were carried out on the test solutions at the Hatano Research Institute. The lowest concentration (3.125 mg/ml) and the highest concentration (50 mg/ml) were sent for analysis. Three test solutions per concentration were stored at room temperature in the dark for 4 hours. The mean analytical concentrations were 103% (low concentration) and 101% (high concentration) of the starting value in the preliminary test, and were 97.1-99.8% (low concentration) and 95.2-98.7% (high concentration). All values were within the permissbale range stipulated by the laboratory, and thaht the test solution was stable for the duration of the test.

Specific details on test material used for the study:

Pentaerythritol (abbreviated to PE in the report), molecular weight 136.2, supplied as a white powder from Japan Chemical Industry Association, manufactured by Mitsubishi Gas Chemical Company. Purity is 92.7% (Lot number: 50825; Containing impurities such as Dipentaerythritol (2.2%), Bis-Pentaerythritol (4.9%), and unknown impurities of (0.2%).

Stability tests were carried out on the test solutions at the Hatano Research Institute. The lowest concentration (3.125 mg/ml) and the highest concentration (50 mg/ml) were sent for analysis. Three test solutions per concentration were stored at room temperature in the dark for 4 hours. The mean analytical concentrations were 103% (low concentration) and 101% (high concentration) of the starting value in the preliminary test, and were 97.1-99.8% (low concentration) and 95.2-98.7% (high concentration). All values were within the permissbale range stipulated by the laboratory, and thaht the test solution was stable for the duration of the test.

Method

Target gene:

Histidine

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

Metabolic activation:

with and without

Metabolic activation system:

S9 mix obtained from the livers of male Sprague-Dawley rats induced with phenobarbitol and 5,6-benzoflavone

Test concentrations with justification for top dose:

Preliminary study: 50 to 5000 µg/plate, with a dose spacing factor of approximately 3.
Main study: 312.5 - 5000 µg/plate, with a dose spacing factor of approximately 2.

Vehicle / solvent:

'Pure water'. 50 mg PE was dissolved per ml of pure water, further dilutions were prepared from this stock solution for immediate use in the experiment.

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

sodium azide

furylfuramide

other: 2-aminoanthracene

Details on test system and experimental conditions:

Bacteria were frozen and stored at < -80°C. At the time of freezing, bacteria were examined for the absence or presence of amino acid requirement, UV radiation sensitivity, membrane mutation (rfa) and ampicillin resistance factor (pKM101). Bacterial cultures were prepared by inoculating the bacteria in an L-shaped test tube containing Nutrient Broth No. 2 (Oxoid), followed by approximately 10 hours incubation at 37°C.

The plate incorporation method was used. 2 ml top agar, 0.1 ml prepared test substance solution, 0.5 ml phosphate-buffered solution or 0.5 ml S9 mix, and 0.1 ml bacteria culture were mixed together in a small test tube and the synthetic culture medium was poured into a flat plate and solidified. In the control group, pure water or the positive control substance solution were used instead of the prepared test substance solution. The medium was cultured at 37°C for 48 hours and the number of mutant colonies counted. Bacterial toxicity was assessed bu examining the bacterial lawn with the naked eye or a microscope.
In the dose-finding study, 3 plates were employed for each of the negative and positive control groups and 1 plate was employed for each respective dose. In the main experiment, 3 plates were employed for each respective dose and for both control groups. The dose-finding study was conducted once and the main experiment was conducted twice.

Evaluation criteria:

The substance was deemed mutagenic if a reproducible doubling or dose-related increase in the number of revertant colonies was seen, compared to negative controls.

Statistics:

No information on statistics is provided in the report.

Results and discussion

Test results

Test results 1

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:

not applicable

Positive controls validity:

valid

Test results 2

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:

not applicable

Positive controls validity:

valid

Test results 3

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:

not applicable

Positive controls validity:

valid

Test results 4

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:

not applicable

Positive controls validity:

valid

Test results 5

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

There was no evidence of cytotoxicity in the preliminary dose-finding study, up to and including the highest concentration of 5000 µg/plate.
In the main study, there were no increases in the number of revertant colonies compared to solvent only controls and no dose-response relationship was evident in the number of revertants (tested up to 5000 µg/plate). This result was confirmed in the independent repeat assay.

All positive and solvent control results were reported to be within the range of historical control data.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

There was no evidence of cytotoxicity in the preliminary dose-finding study, up to and including the highest concentration of 5000 µg/plate. In the main study, there were no increases in the number of revertant colonies compared to solvent only controls and no dose-respone relationship was evident in the number of revertants (tested up to 5000 µg/plate). This result was confirmed in the independent repeat assay.

All positive and solvent control results were reported to be within the range of historical control data.

Applicant's summary and conclusion

Conclusions:

Pentaerythritol was found to be negative in the Ames test with and without metabolic activation, tested up to a concentration of 5000 µg/plate.

Executive summary:

The mutagenic potential of Pentaerythritol was determined in the bacterial reverse mutation assay (Ames test). Salmonella typhimurium strains TA100, TA1535, TA1537 and TA98 were tested along with Escherichia coli strain WP2 urvA in both the presence and absence of metabolic activation (S9 mix). A preliminary cytoxicity test was conducted, in which no evidence of bacterial toxicity was seen up to and including concentrations of 5000 µg/plate. In the main study, there were no increases in the number of revertant colonies compared to solvent only controls and no dose-respone relationship was evident in the number of revertants (tested up to 5000 µg/plate). This result was confirmed in the independent repeat assay.

No evidence of mutagenicity was seen under the conditions of this assay.