Reaction product of a ((2S)-2, 6-diaminohexanoic acid hydrochloride salt, phosphonic acid and methanal) with a 1,2-disubstituted alkane and sodium hydroxide.

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to a test protocol that is comparable to the appropriate OECD test guideline, with acceptable restrictions. The restrictions were that only four strains of bacteria were used.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1981

Report date:

1981

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

insufficient range of strains to meet requirements of current guideline

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Details on test material:

Dequest 2000. Indicated as 50% ATMP in water on sample identification in report. Specific gravity: 1.3. Dosed as supplied. Concentrations in this entry converted by reviewer to µg ATMP/ml on basis of this information.

Method

Species / strain

Species / strain / cell type:

other: Salmonella typhimurium TA98, 100, 1535, 1537

Metabolic activation:

with and without

Metabolic activation system:

Aroclor induced mouse and rat liver S9

Test concentrations with justification for top dose:

0.01, 0.04, 0.2, 1, 3,10ul Dequest 2000/plate and 25ul Dequest 2000 Dequest/plate for spot test. Expressed as ATMP: 0.0065, 0.026, 0.13, 0.65, 1.95, 6.5ul/plate for plate incorporation; up to 9ul for spot test

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: water
- Justification for choice of solvent/vehicle: none given

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

water

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

TA 98 and TA 100 without activation

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: sodium nitrite

Remarks:

TA 1535 without activation

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

TA 1537 without activation

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Remarks:

TA 98 and TA 100 with activation

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Remarks:

TA 1535 and TA 1537 with activation

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium; in agar (plate incorporation); as impregnation on paper disk


DURATION
- Preincubation period: none
- Exposure duration: 48 hours
- Expression time (cells in growth medium): 48 hours
- Fixation time (start of exposure up to fixation or harvest of cells): 48 hours

NUMBER OF REPLICATIONS: single applications (spot test), triplicate plates (plate incorporation)

DETERMINATION OF CYTOTOXICITY
- Method: other: reduction in microbial lawn

Evaluation criteria:

A positive response is indicated if three of more treatments are significantly greater than the solvent control with a significant dose-response.

Statistics:

Analysis of log10 revertants per plate used Bartlett's test for homogeneity of variance and comparison of treatments with controls used within-levels pooled variance and a one-sided t-test. Where values were found to be significant, a Grubb's test was applied and significance of dose-response was evaluated by a t-test.

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:

cytotoxicity

Remarks:

1-3ul/plate for plate incorporation

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

ADDITIONAL INFORMATION ON CYTOTOXICITY: In the main study toxicity was seen as follows: +S9; 10ul, 3ul (TA98, TA1535); 10ul, 3ul, 1ul (TA100, TA1537) -S9: 10ul, 3ul (all strains)

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

No increase in revertants in any strain/metabolic activation condition. Full details of results presented in report.  Positive and negative values acceptable.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative with and without activation

Dequest 2000 showed a lack of mutagenic potential when tested in 4 strains of Salmonella in the absence and presence of rat and mouse liver S9 mix up to the toxicity limit (1-3ul/plate). It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test.