1,4-dithiane-2,5-diol

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2004-06-30 to 2004-08-09

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

21 July 1997

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2000

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

1,4-dithiane-2,5-diol

EC Number:

254-751-6

EC Name:

1,4-dithiane-2,5-diol

Cas Number:

40018-26-6

Molecular formula:

C4H8O2S2

IUPAC Name:

1,4-dithiane-2,5-diol

Test material form:

solid: particulate/powder

Method

Target gene:

his-; uvrB-; WP2uvrA-

Species / strain

Species / strain 1

Species / strain / cell type:

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

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

S9 Mix

Test concentrations with justification for top dose:

Preliminary Test: 0, 0.15, 0.5, 1.5, 5, 15, 50, 150, 500, 1500 and 5000 µg/plate.
Main Test:
Salmonella without S9-mix: 1,5, 5, 15, 50, 150, 500 and 1500 µg/plate.
Salmonella with S9-mix (except TA1535), E. coli, with and without S9-mix: 5, 15, 50, 150, 500 and 1500 µg/plate.
TA1535 (with S9-mix only): 15, 50, 1513, 500, 1500 and 5000 µg/plate.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: due to insolubility of the test substance in water

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

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: 2-Aminoanthracene; with metabolic activation

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium; in agar (plate incorporation)
- Cell density at seeding (if applicable): 1 to 9.9E+006 bacteria per mL

DURATION
- Exposure duration: 48h

NUMBER OF REPLICATIONS: 3 (test concentrations, vehicle control and positive controls)

DETERMINATION OF CYTOTOXICITY
- Method: decrease in the number of revertant colonies per plate and/or thinning or disappearance of the background bacterial lawn, or appearance of pinhead colonies in treated cultures.
- Any supplementary information relevant to cytotoxicity: The test material caused a visible reduction in the growth ef the bacterial background lawn to all of the strains, initially at 500 and 150 µg/plate, with and without S9-mix, respectively.

Rationale for test conditions:

Tested up to the toxic limit or, in case of TA1535, experiment 1 only the maximum recommended dose level of 5000 µg/plate (in the presence of S9 only).

Evaluation criteria:

The test material may be considered positive in this test system if the following criteria are met:
The test material should have induced a reproducible, dose-related and statistically (Dunnett´s method of linear regression) significant increase in the revertant count in at least one strain of bacteria.

Results and discussion

Test results

Key result

Species / strain:

other: TA98, TA100, TA1535, TA1537; E.coli WP2uvrA-

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Any other information on results incl. tables

Table 1: Number of revertants per plate (mean of 3 plates)

Experiment 1:

	[TA100]			[TA1535]			[WP2uvrA-]			[TA98]			[TA1537]
Conc. [unit]	— MA	+ MA	Cytotoxic (yes/no)	— MA	+ MA	Cytotoxic (yes/no)	— MA	+ MA	Cytotoxic (yes/no)	— MA	+ MA	Cytotoxic (yes/no)	— MA	+ MA	Cytotoxic (yes/no)
0*	87	83	no	30	14	no	19	26	no	15	23	no	15	19	no
1.5	77	75	no	39	N/T	no	N/T	28	no	14	22	no	12	21	no
5	71	86	no	44	9	no	19	27	no	17	27	no	9	18	no
15	79	78	no	44	14	no	21	25	no	14	26	no	11	18	no
50	65	88	no	29	10	no	20	28	no	13	31	no	6	21	no
150	42	92	yes	23	17	no	21	19	no	16	27	yes	9	15	no
500	0	0	yes	0	0	yes	16	0	yes	0	0	yes	0	0	yes
1500	0	N/T	yes	0	0	yes	0	N/T	yes	0	N/T	yes	0	N/T	yes
Positive control	465	948		441	294		716	238		158	181		979	193

Experiment 2:

	[TA100]			[TA1535]			[WP2uvrA-]			[TA98]			[TA1537]
Conc. [µg/plate]	— MA	+ MA	Cytotoxic (yes/no)	— MA	+ MA	Cytotoxic (yes/no)	— MA	+ MA	Cytotoxic (yes/no)	— MA	+ MA	Cytotoxic (yes/no)	— MA	+ MA	Cytotoxic (yes/no)
0*	101	93	no	31	15	no	27	35	no	18	28	no	11	22	no
0.5	102		no	35		no	N/T		no	25		no	11		no
1.5	100		no	32		no	N/T		no	24		no	15		no
5	113	94	no	34	15	no	21	35	no	16	27	no	18	20	no
15	103	85	no	29	16	no	21	27	no	18	32	no	15	19	no
50	93	94	no	29	10	no	24	26	no	18	34	no	15	23	no
150	94	87	no	32	17	no	22	33	no	18	31	no	15	20	no
500	0	81	yes	0	22	yes	27	23	no	0	29	yes	0	20	yes
1500	N/T	0	yes	N/T	0	yes	0	0	yes	N/T	3	yes	N/T	0	yes
Positive control	385	826		211	248		789	332		177	228		1020	308

Applicant's summary and conclusion

Conclusions:

In the present study according to OECD guideline 471 (adopted 21 July 1997) four Salmonella typhimurium strains and one Escherichia coli strain were incubated with the test substance at the following concentrations: 0, 0.15, 0.5, 1.5, 5, 15, 50, 150, 500, 1500, and 5000 µg/plate. Since the validity criteria of the test were fulfilled and there were no increases in the revertant count up to cytotoxic concentrations, the substance is considered to be non-mutagenic under the described test conditions.Thus, the test substance does not nedd to be classified according to Regulation (EC) No 1272/2008 (CLP) and the Globally Harmonized System for Classification and Labelling of Chemicals (GHS) with respect to mutagenicity.