1,2,3-Oxadiazolium, 5-amino-3-(4-morpholinyl)-, chloride (1:1)

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: genome mutation

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline and GLP-Study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1994

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: Safepharm Standard Method OECD 26F

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

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

Test guideline 3

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

5-amino-3-(morpholin-4-yl)-1,2,3-oxadiazolidin-3-ium chloride

EC Number:

605-254-1

Cas Number:

16142-27-1

Molecular formula:

C6 H11 N4 O2 . Cl

IUPAC Name:

5-amino-3-(morpholin-4-yl)-1,2,3-oxadiazolidin-3-ium chloride

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): Linsodomin Substanz
- Physical state: white powder
- Lot/batch No.: W003
- Storage condition of test material: brown glass bottle, room temperature in the dark

Method

Species / strain

Species / strain / cell type:

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

Metabolic activation:

with and without

Metabolic activation system:

10 % liver S9 in standard co-factors

Test concentrations with justification for top dose:

0, 312.5, 625, 1250, 2500, 5000 µg/Plate (preliminary toxicity study)
0, 8, 40, 200, 1000, 5000 µg/Plate (Experiment 1)
0, 312.5, 625, 1250, 2500, 5000 µg/Plate (Experiment 2)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: ethanol

Controls

Untreated negative controls:

yes

Remarks:

solvent treatment group

Negative solvent / vehicle controls:

yes

Remarks:

negative controll group

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

9-aminoacridine

N-ethyl-N-nitro-N-nitrosoguanidine

benzo(a)pyrene

other: 4-Nitro-o-phenylenediamine and 2-Aminoanthracene

Details on test system and experimental conditions:

The tester strains were obtained from the British Industrial Biological Research Association and were stored at -196°C in a Statebourne liquid nitrogen freezer. Prior to being useed, characterization checks were carried out to determine the amino acid requirement, presence of rfa, R factors, uvr mutation and the spontaneous reversion rate. Overnight subcultures of the appropriate coded stock cultures were prepared in nutrient broth (Origin and Lot given in report) and incubated at 37°C for approximately 10 hours.
The microsomal enzyme fraction (Origin and Lot given in report) was prepared from the liver of male Sprague-Dawley rats weighing ~200 g. These had received a single i.p. injection of Aroclor 1254 at 500 mg/kg, 5 days before S9 preparation.

A prliminary toxicity study was carried out to determine the toxicity of the test material to the tester organisms.

The mutation studies were conducted using the direct plate incorporation method in accordance with the standard method for mutagenicity tests using bacteria. Five concentrations of the test material were assayed in triplicate against each tester strain.

Evaluation criteria:

To be considered negative the number of induced revertants compared to spontaneous revertants should be less then twofold on each dose level employed, the intervals of which should be between 2 and 5 fold and extend to the limits imposed by toxicity, solubility or up to the maximum recommended dose of 5000 µg/plate. In this case the limiting factor was the maximum recommended dose.

Statistics:

All data are statistically analysed using the methods recommended by the UKEMS (Kirkland, D.J. (Ed) Statistical Evaluation of Mutagenicity Test Data. UKEMS Sub-committee on Guidelines for Mutagenicity Testing. Report- Part III (1998) Cambridge University Press) and normally Dunnett´s method of linear regression is used to evaluate the results.

Results and discussion

Test results

Test results 1

Species / strain:

other: TA1535 and TA1537

Metabolic activation:

with and without

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
positive

A statistically significant, dose related and reproducible increase in the number of revertant colonies of bacteria was recorded for the salmonella strains TA1535 and TA1537. A much smaller response was observed for TA100 in th esecond experiment only.
Linsidomin was found to be mutagenic under the conditions of this test.