2-Propenal, 2-fluoro-3-(4-morpholinyl)-, (Z)-

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

The mutagenic potential of the test material was evaluated in a Salmonella/microsome test with the S. typhimurium strains TA 98, TA 100, TA 102, TA 1535 and TA 1537 in the presence and absence of S9 mix according to OECD TG 471. Doses up to and including 5000 µg per plate did not cause any bacteriotoxic effects. Total bacteria counts remained unchanged and no inhibition of growth was observed.Test item precipitation did not occur. Historical negative control data demonstrated that no deleterious or mutagenic effects were induced by the chosen solvent. The positive controls increased mutant counts to well over those of the solvent controls, and thus demonstrated the system's sensitivity and the activity of the S9 mix. None of the five strains used showed a dose-related and biologically relevant increase in mutant counts over those of the solvent controls in the preincubation test. This applied both to the tests with and without S9 mix. Due to these results the test item has to be regarded as non-mutagenic.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

February-March 2016

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

method of treatment is restricted to preincubation

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

method of treatment is restricted to preincubation

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: refrigerator, approx. 4°C
- Solubility and stability of the test substance in the solvent/vehicle: The test item is an impurity of a drug substance. Thus, according to ICH Draft Consensus Guideline M7, no stability test in the solvent was performed.


TREATMENT OF TEST MATERIAL PRIOR TO TESTING

- Final preparation of a solid: solution (only used at the same day) - The purity of the test item was not taken into account for the calculation of the dosages.

Target gene:

Histidine gene locus

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 induced male Sprague Dawley rat liver S9 mix

Test concentrations with justification for top dose:

0, 100, 250, 500, 1000, 2500, 5000 µg/plate (+/-S9 mix, all strains)

Vehicle / solvent:

DMSO

Untreated negative controls:

yes

Negative solvent / vehicle controls:

no

Remarks:

No solvent control was used since sufficient evidence was available in the literature and from testing laboratory experience, indicating that the solvents used had no influence on the spontaneous mutant counts of the used strains.

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: sodium azide (TA 1535, TA 100), 4-nitro-1,2-phenylene diamine (TA 1537), 2-Nitrofluoren (TA 98), Cumene hydroperoxide (TA 102), 2-aminoanthracene (all strains).

Details on test system and experimental conditions:

METHOD: each concentration including the controls was tested in triplicate.

Evaluation criteria:

A reproducible and dose-related increase in mutant counts of at least one strain is considered to be a positive result. For TA1535, TA100, TA1537 and TA98 this increase should be about twofold as compared to the solvent controls. For TA102 an increase of about 100 mutants should be reached. Otherwise, the result is evaluated as negative. However, these criteria may be overruled by good scientific judgment. In case of questionable results, investigations should continue, possibly with modifications, until a final evaluation is possible.

Species / strain:

S. typhimurium, other: TA1535, TA100, TA1537, TA98 and TA102

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:

not examined

Untreated negative controls validity:

valid

Positive controls validity:

valid

Table 1: Summary of results from the Salmonella mutagenicity assay with FMA (mean values of revertants per plate)

Dose (µg per plate)	Without metabolic activation
	TA 1535	TA 100	TA 1537	TA 98	TA 102
0	9	165	6	17	213
100	7	160	7	15	208
250	8	166	8	17	216
500	8	167	7	19	212
1000	9	155	8	17	206
2500	9	160	7	18	234
5000	12	170	8	15	219
Positive control	685	1274	55	1405	505
Dose ( µg per plate )	With metabolic activation (liver S9 mix)
	TA 1535	TA 100	TA 1537	TA 98	TA 102
0	9	210	8	27	294
100	9	212	8	26	240
250	8	224	8	28	277
500	11	251	8	25	279
1000	8	193	7	22	289
2500	10	177	10	25	296
5000	11	177	9	26	277
Positive control	153	3021	241	2270	853

Historical negative control data demonstrated that no deleterious or mutagenic effects were induced by the chosen solvent. The positive controls sodium azide, 4-nitro-1,2 -phenylene diamine, 2-nitrofluoren, cumene hydroperoxide and 2-aminoanthracene increased mutant counts to well over those of the solvent controls, and thus demonstrated the system's sensitivity and the activity of the S9 mix.

None of the five strains used showed a dose-related and biologically relevant increase inmutant counts over those of the solvent controls in the preincubation test. This applied both to the tests with and without S9 mix.

 

 

Conclusions:

negative

Executive summary:

The mutagenic potential of the test material was evaluated in a Salmonella/microsome test with the S. typhimurium strains TA 98, TA 100, TA 102, TA 1535 and TA 1537 in the presence and absence of S9 mix according to OECD TG 471. Doses up to and including 5000 µg per plate did not cause any bacteriotoxic effects. Total bacteria counts remained unchanged and no inhibition of growth was observed. Test item precipitation did not occur. Historical negative control data demonstrated that no deleterious or mutagenic effects were induced by the chosen solvent. The positive controls increased mutant counts to well over those of the solvent controls, and thus demonstrated the system's sensitivity and the activity of the S9 mix. None of the five strains used showed a dose-related and biologically relevant increase in mutant counts over those of the solvent controls in the preincubation test. This applied both to the tests with and without S9 mix. Due to these results the test item has to be regarded as non-mutagenic.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Based on the available information classification according to Regulation (EC) No. 1272/2008 (CLP) is not required.