1,3-dihydroxyacetone

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:

1983-07-06 to 1983-08-17

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1983

Report date:

1983

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted 1983

Deviations:

yes

Remarks:

The highest dose tested was 10,000 µg/plate and, thus, exceeding the maximum test concentration of 5,000 µg/plate as recommended by the Guideline.

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

The highest dose tested was 10,000 µg/plate and, thus, exceeding the maximum test concentration of 5,000 µg/plate as recommended by the Guideline.

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

1,3-dihydroxyacetone

EC Number:

202-494-5

EC Name:

1,3-dihydroxyacetone

Cas Number:

96-26-4

Molecular formula:

C3H6O3

IUPAC Name:

1,3-dihydroxyacetone

Details on test material:

Purity: 99.0 %

Method

Target gene:

His Operon

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 98

Details on mammalian cell type (if applicable):

his D 3052, uvrB, rfa + R-factor (pKM101)

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 100

Details on mammalian cell type (if applicable):

his G 46, uvrB, rfa + R-factor (pKM101)

Species / strain 3

Species / strain / cell type:

S. typhimurium TA 102

Details on mammalian cell type (if applicable):

his G428, rfa + R-factor (pKM101)

Species / strain 4

Species / strain / cell type:

S. typhimurium TA 1535

Details on mammalian cell type (if applicable):

his G 46, uvrB, rfa

Species / strain 5

Species / strain / cell type:

S. typhimurium TA 1537

Details on mammalian cell type (if applicable):

his C 3076, uvrB, rfa

Metabolic activation:

with and without

Metabolic activation system:

10% rat liver homogenate (S9 mix) with standard co-factors with metabolic activation (Aroclor)

Test concentrations with justification for top dose:

1st and 2nd test: 50, 250, 1250, 2500, 5000, and 10000 µg/plate

Vehicle / solvent:

DMSO

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

other: 2-Aminoanthracene

Remarks:

with S9

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

N-ethyl-N-nitro-N-nitrosoguanidine

methylmethanesulfonate

mitomycin C

other: N-Methyl-N-nitro-N-nitrososguanidine

Details on test system and experimental conditions:

NUMBER OF REPLICATIONS:
- Number of cultures per concentration: 4 plates per test concentrations, 8 (2x 4) plates per solvent control
- Number of independent experiments: 2

METHOD OF TREATMENT/ EXPOSURE:
- Test substance added in agar (plate incorporation)

Evaluation criteria:

No evaluation criteria are provided in the original report. For the reevaluation of the results of this study the following criteria are taken into account:
A test material was to be defined as positive or mutagenic in this assay if
- a biologically relevant increase in the mean number of revertants above a threshold of 2-fold (TA 98, TA 100, TA 102) or 3-fold (TA 1535, TA 1537) as compared to the concurrent negative controls is observed.
- an increase exceeding the threshold at only one concentration is considered as biologically meaning ful if reproduced in a second independent experiment
- a concentration-dependent increase is considered biologically meaningful if the threshold is exceeded at more than one concentration

Statistics:

no

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 98

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

Test results 2

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

positive

Remarks:

at concentrations larger than 2500 µg/plate

Cytotoxicity / choice of top concentrations:

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

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium 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

Test results 4

Key result

Species / strain:

S. typhimurium TA 1535

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

Test results 5

Key result

Species / strain:

S. typhimurium TA 1537

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

Any other information on results incl. tables

The test material was examined for mutagenic activity in two series of in vitro microbial assays employing Salmonella typhimurium indicator organisms. The substance was tested with and without the presence of a metabolic activation system, i.e. liver microsomal enzyme preparations from Aroclor-induced rats.

The test material was tested over a series of concentrations. The choice of the concentration range was mainly influenced by the endeavor to achieve a toxic effect in the highest concentration. Such a toxic effect is generally considered important in this test system. This effect was not obtained in the present trials: No toxicity and no precipitation were observed up to the highest dose tested at both experiments.

During the routinely conducted microscopic examination of the plates no toxic effect was detected. The dose range employed for the evaluation of this compound was from 50 to 10000µg per plate, i.e. the highest dose exceeded the maximum test material concentration as recommended by the OECD guideline.

The results of the tests conducted on the test material in the absence and presence of a metabolic system were negative for Salmonella typh. TA 98, TA 102, TA 1535, and TA 1537. The repeat tests were also negative.

With the tester strains Salmonella typhimurium TA 100 the observed increase in the number of revertant colonies was interpreted as a positive result.

The validity of the mutation assay can be assessed by the results obtained for the positive and negative controls. The negative control mutant frequencies were all in the normal range, and the positive control compounds yielded normal mutant frequencies that were greatly in excess of the background.

Applicant's summary and conclusion

Conclusions:

Based on the results of this study, the test material is considered to be weakly positive in this bacterial reverse mutation assay.

Executive summary:

The investigations were performed using Salmonella typhimurium TA 100, TA 102, TA 98, TA 1535 and TA 1537 as tester strains. The mutagenic potential was examined in the plate incorporation test with and without addition of a liver postmitochondrial fraction as the in vitro metabolizing system (S-9; male rats, induced with Aroclor 1254).

Concentrations: 50, 250, 1250, 2500, 5000, and 10000 µg/plate.

2-Aminoanthracene, 9-aminoacridine, benzo(a)pyrene, daunomycin, 1-ethyl-2-nitro-3-nitrosoguanidine, methyl methanesulfonate, N-methyl-N-nitro-N-nitro­soguanidine, mitomycin c, and 2-nitrofluorene served as positive controls for testing the bacteria and the induced S-9 preparation.

With and without addition of S-9 as the metabolizing system, the test material did show mutagenic activity with Salmonella typhimurium TA 100 in the concentration range used. With Salmonella typhimurium TA 98, TA 102, TA 1535 and TA 1537 no mutagnic activity was found. With Salmonella typhimurium TA 100 the number of revertant colonies was increased 2.4 - 3.8 -fold. The maximum concentration level in this study exceeded the highest test concentration as recommended by the current OECD-Guideline for this system. Toxicity and precipitation was not observed up to the highest dose tested.

The substances used as positive controls showed normal reversion properties of all strains and good metabolic activity of the S-9 mix used.