2-phosphonobutane-1,2,4-tricarboxylic 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

Study period:

1979-08

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Only four instead of at least five strains of bacteria were used.

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1979

Report date:

1979

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted 26 May 1983

Deviations:

yes

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

Directive 84/449/EEC

Deviations:

yes

Principles of method if other than guideline:

Only four instead of at least five strains of bacteria were used. It has to be mentioned that the test was performed in 1979, the current guideline was adopted in 1997.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2-phosphonobutane-1,2,4-tricarboxylic acid

EC Number:

253-733-5

EC Name:

2-phosphonobutane-1,2,4-tricarboxylic acid

Cas Number:

37971-36-1

Molecular formula:

C7H11O9P

IUPAC Name:

2-phosphonobutane-1,2,4-tricarboxylic acid

Details on test material:

- technical pure product (category AC)
- 45-50% (w/w) 2-phosphonobutane-1,2,4-tricarboxylic acid in water
- Pt. 10 E 2806

Method

Target gene:

The Salmonella typhimurium strains are constructed to differentiate between base pair (TA 1535, TA 100) and frameshift (TA 1537, TA 98) mutations. All stains show increased sensitivity due to "deep rough" (one polysaccharide chain is missing, hence, also large molecules can penetrate into the bacteria cells ) and UV-repair-deficiency.

Species / strain

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

NA

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

S9

Test concentrations with justification for top dose:

nominal concentrations: 20; 100; 500; 2500; 12,500 µg /plate

Vehicle / solvent:

- DMSO (BAHIBIT AM; Trypaflavin)
- demineralised water (Endoxan)

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

0 µg BAHIBIT AM /plate / demineralised water

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

cyclophosphamide

Remarks:

Substance name: Endoxan; cytostatic agent; the active agent cyclophosphamid is a well-known pro-mutagen; 217. 5 µg equivalent to 150 µg cyclophosphamid; only for strains 1535 and TA 100

Controls 2

Negative solvent / vehicle controls:

yes

Remarks:

demineralised water

Positive controls:

yes

Positive control substance:

other: Trypaflavin

Remarks:

Active substance of Panflavin and Rivanols; pro-mutagen for frameshift mutations; 200 µg; only for strains 1537 and TA 98

Details on test system and experimental conditions:

Incubation for 48h at 37°C.
4 plates / substance / strain
Negative controls:
4 plates / DMSO / strain
4 plates / demineralised water / stain
Additionally, the total germ account was determined for 2 plates/group.

Evaluation criteria:

A positive result requires a reproducible dose-related duplication of mutants compared with the negative controls for at least one strain.
The colony numbers for negative controls, positive controls and the test plates were determined, the mean values were calculated.

Statistics:

Mean values

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:

other: only weak bacteriotoxic effects for concentrations >500 µg/ plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

NA

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

No remarks.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

BAYHIBIT AM containing 45-50% (w/w) 2-phosphonobutane-1,2,4-tricarboxylic acid in water was found to be non-mutagenic under test conditions.

Executive summary:

The Ames test was performed with BAYHIBIT AM containing 45-50% (w/w) 2-phosphonobutane-1,2,4-tricarboxylic acid in water, at nominal concentrations of 16; 80; 400; 2500 and 12,500 µg /plate. Strain-specific bacteriotoxicity was observed at doses >500 µg/ plate and higher. The reported data of this mutagenicity assay shows, that under the experimental conditions reported, the test item did not induce gene mutations by frameshift or base-pair substitution strains used. Hence, BAYHIBIT AM containing 45-50% (w/w) 2-phosphonobutane-1,2,4-tricarboxylic acid is considered non-mutagenic in this bacterial reverse mutation assay.