Diammonium hydrogen 2-hydroxypropane-1,2,3-tricarboxylate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

not specified

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Remarks:

Significant experimental and reporting deficiencies: purity/stability of test item missing; method was not fully described (incubation duration & temperature missing; evaluation criteria missing; treatment procedure missing); individual data missing; historical control data was missing; three concentrations were only tested instead of five concentrations; information on cytotoxicity indicates that higher concentrations could have been tested; 2-aminoanthracene was used during testing without metabolic activation; confirmatory experiment missing

Data source

Reference

Reference Type:

publication

Title:

Absence of mutagenic activity of acidity regulators in the Ames Salmonella/microsome test

Author:

Al-Ani, F. Y. & Al-Lami, S. K.

Year:

1988

Bibliographic source:

Mutation Research 206: 467 - 470.

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

1983-05-26

Deviations:

yes

Remarks:

please refer to the field "Rationale for reliability incl. deficiencies" above

GLP compliance:

not specified

Remarks:

not specified in the publication

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Citric acid

EC Number:

201-069-1

EC Name:

Citric acid

Cas Number:

77-92-9

Molecular formula:

C6H8O7

Test material form:

not specified

Details on test material:

- Source: State enterprise for dairy products

Specific details on test material used for the study:

not specified

Method

Target gene:

TA97: his D 6610 his O 1242
TA98: his D 3052
TA100: his G 46

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA97, TA98, TA100 and TA104

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

500, 1000 and 2000 µg/plate (with and without metabolic activation)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: distilled water

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

distilled water

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium; in agar (plate incorporation)

NUMBER OF REPLICATIONS: triplicates

DETERMINATION OF CYTOTOXICITY
- Method: bacterial background lawn growth

Rationale for test conditions:

not specified

Evaluation criteria:

not specified

Statistics:

means

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA97, TA98, TA100 and TA104

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Additional information on results:

The spontaneous reversions of the tester strains were within the acceptable range (Levin et al., 1982a,b; Maron and Ames, 1983; Levin and Ames, 1986). A decreased spontaneous reversion frequency of TA97 was noted.

In the presence or absence of S9 mix, the numbers of revertants in all tester strains for all concentrations of the test item tested were not significantly different from the respective negative controls. In some cases the number of revertants with S9 was statistically higher than the number of colonies on negative control plates.

Routine examination of the bacterial background lawn indicates the absence of toxicity associated with the doses of the compounds tested.

*References:
- Levin, D.E., E. Yamasaki and B.N. Ames (1982a) A new Salmonella tester strain TA97, for the detection of frameshift mutagens. A run of cytosines as a mutational hot-spot, Mutation Res., 94, 315 330.
- Levin, D.E., M.C. Hollstein, M.F. Christman, E.A. Schwiers and B.N. Ames (1982b) A new Salmonella tester strain, TA102, with A:T base pairs at the site of mutation to detect oxidative mutagens, Proc. Natl. Acad. Sci. (U.S.A.), 79, 7445-7449.
- Maron, D.M., and B.N. Ames (1983) Revised methods for the Salmonella mutagenicity test, Mutation Res., 113, 173-215.
- Levin, D.E., and B.N. Ames (1986) Classifying mutagens as to their specificity in causing the six possible transitions and transversions: a simple analysis using the Salmonella mutagenicity
assay, Environ. Mutagen., 8, 9 28.

Applicant's summary and conclusion

Conclusions:

According to the authors, the substance tested non-mutagenic under the conditions of the study.