Aspartic acid

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

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

1. The test substance is commercially available and was characterized by the supplier prior to the initiation of this study. A certificate of analysis was provided. Although the characterization was not performed under Good Laboratory Practice Standards,

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Aspartic acid

EC Number:

200-291-6

EC Name:

Aspartic acid

Cas Number:

56-84-8

Molecular formula:

C4H7NO4

IUPAC Name:

aspartic acid

Details on test material:

Substance Tested: ¿ L-Asparate
¿ 56-84-8 (CAS Number)
¿ L-Aspartic Acid
Haskell Number: 27761
Composition: Not supplied by the sponsor
Purity: ¿ 99.5%
Physical Characteristics: White solid
Stability: The test substance appeared to be stable under the conditions of the study; no evidence of instability was observed.

Method

Target gene:

Tester strain cultures were checked for the following genetic markers on the day of the
preparation of master plates.
The histidine requirement was tested by comparing the growth of each Salmonella tester strain
on a histidine/biotin-supplemented minimum glucose agar plate with their growth on a biotinonly
minimum glucose agar plate.
The tryptophan requirement was tested by comparing the growth of WP2uvrA strain on a
tryptophan-supplemented minimum glucose agar plate with their growth on a minimum glucose
agar plate.
For the Salmonella tester strains the presence of the rfa wall mutation was confirmed by
demonstration of the sensitivity of the cultures to crystal violet.
The presence of uvrA and uvrB mutation was demonstrated by the sensitivity to ultraviolet light
of the tester strains.
The presence of the pKM101 plasmid was confirmed for cultures of tester strains TA98 and
TA100 by demonstration of resistance to ampicillin

Species / strain

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

The specific genotype characteristics of these strains are as follows:
Additional Mutations
Tester Strain HIS/Trp Mutation Repair LPS Plasmid
S. typhimurium TA98 hisD3052 ¿uvrB rfa pkM101
S. typhimurium TA100 hisG46 ¿uvrB rfa pkM101
S. typhimurium TA1535 hisG46 ¿uvrB rfa --
S. typhimurium TA1537 hisC3076 ¿uvrB rfa --
Escherichia coli WP2uvrA trpE ¿uvrA - --

Metabolic activation:

with and without

Metabolic activation system:

Liver homogenate (S9, average protein concentration: 35.8 mg/mL) prepared from male Sprague-Dawley rats induced with Aroclor 1254 was purchased commercially (Moltox Inc., Boone, North Carolina, U.S.A.).

Test concentrations with justification for top dose:

The dose levels for the toxicity-mutation test were 33.3, 66.7, 100, 333, 667, 1000, 3333, and 5000 ¿g per plate. The dose levels for the mutagenicity test were 333, 667, 1000, 3333, and 5000 ¿g per plate

Controls

Untreated negative controls:

yes

Remarks:

sterile water (CAS 7732-18-5, molecular grade, Mediatech Inc.)

Positive controls:

yes

Remarks:

benzo[a]pyrene (CAS 50-32-8) 4-nitroquinoline N-oxide (CAS 56-57-5) acridine mutagen ICR-191 (CAS 17070-45-0) sodium azide (CAS 26628-22-8) 2-aminoanthracene (CAS 613-13-8) 2-nitrofluorene (CAS 607-57-8)

Remarks:

The positive controls were dissolved in dimethyl sulfoxide (DMSO, CAS 67-68-6, 99.9% purity, EMD), except for sodium azide and ICR-191, which were dissolved in sterile water.

Results and discussion

Test results

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Toxicity-Mutation Test

In the toxicity-mutation test, the maximum dose evaluated was 5000 ¿g/plate for tester strains TA98, TA100, TA1535, TA1537, and WP2uvrA in the absence and presence of S9 metabolic activation. This dose was achieved using a concentration of 50 mg/mL and a 100 ¿L plating aliquot. The dose levels used in this test were 33.3, 66.7, 100, 333, 667, 1000, 3333, and 5000 ¿g/plate. The plate incorporation method was employed. No positive mutagenic responses were observed at any dose level in any tester strain in the absence or presence of S9 metabolic activation. No toxicity was observed at any dose level with any tester strain in either the absence or presence of S9 activation. No test substance precipitation was observed at any dose level with any tester strain in either the absence or presence of S9 metabolic activation.

Mutagenicity Test

Based on the toxicity-mutation test, the maximum dose evaluated in the mutagenicity test was 5000 ¿g/plate for tester strains TA98, TA100, TA1535, TA1537, and WP2uvrA in the absence and presence of S9 metabolic activation. This dose was achieved using a concentration of 50 mg/mL and a 100 ¿L plating aliquot. The dose levels used in this test were 333, 667, 1000, 3333, and 5000 ¿g/plate for all tester strains. The plate incorporation method was employed. No positive mutagenic responses were observed at any dose level or with any tester strain in either the absence or presence of S9 metabolic activation. No toxicity or test substance precipitation was observed at any dose level with any tester strain in either the absence or presence of S9 metabolic activation.