Methoxycyclododecane

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20 Oct - 21 Nov 2000

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2000

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Ministerium für Umwelt und Verkehr, Baden-Württemberg, Germany

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Methoxycyclododecane

EC Number:

221-053-8

EC Name:

Methoxycyclododecane

Cas Number:

2986-54-1

Molecular formula:

C13H26O

IUPAC Name:

methoxycyclododecane

Method

Target gene:

his operon

Species / strain

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:

co-factor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Aroclor 1254

Test concentrations with justification for top dose:

First experiment:
15; 50; 150; 500; 1500 and 5000 µg/plate with and without metabolic activation

Second experiment:
5; 15; 50; 150; 500; 1500 and 5000 µg/plate with and without metabolic activation

Vehicle / solvent:

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

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

mitomycin C

other: 2-aminoanthracene

Remarks:

+S9: 2-AA (0.8µg/plate for TA98, TA100, TA102, TA1535; 1.7 µg/plate for TA1537); -S9: NaN3 (0.7 µg/plate for TA100, TA1535), 2-NF (2.5 µg/plate for TA98), 9-AA (50 µg/plate for TA1537), Mitomycin C (0.15 µg/plate for TA102)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 to 72 h

NUMBER OF REPLICATIONS: triplicates each in two independent experiments

DETERMINATION OF CYTOTOXICITY
- Method: Inspection of the bacterial background lawn

Statistics:

Mean values and standard deviation were calculated.

Results and discussion

Test results

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 1500 µg/plate (TA100, -S9 and +S9; TA102, +S9))

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: Precipitation of the test compound was observed at 5000 µg/plate

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Test Results of Experiment 1

EXPERIMENT 1
S9-Mix	Without
Test item (µg/plate)	TA 98	TA 100	TA 102	TA 1535	TA 1537
NC	15 ± 4	99 ± 6	288 ± 19	19 ± 6	12 ± 2
Solvent control (ethanol)	20 ± 6	88 ± 3	281 ± 3	20 ± 5	10 ± 1
15	-	-	288 ± 19	19 ± 2	-
50	23 ± 1	87 ± 19	259 ± 24	16 ± 3	7 ± 3
150	17 ± 5	85 ± 10	222 ± 12	22 ± 8	8 ± 5
500	15 ± 2	86 ± 14	227 ± 23	15 ± 2	10 ± 5
1500	17 ± 5	73 ± 10 (T)	230 ± 4	17 ± 5	7 ± 2
5000 (P)	20 ± 5	76 ± 6 (T)	153 ± 47 (T)	19 ± 2	6 ± 3 (T)
NaN3 (0.7)	-	328 ± 17	-	295 ± 6	-
2-NF (2.5)	227 ± 17	-	-	-	-
9-AA (50)	-	-	-	-	358 ± 77
Mitomycin C (0.15)	-	-	687 ± 65	-	-
S9-Mix	With
Test item (µg/plate)	TA 98	TA 100	TA 102	TA 1535	TA 1537
NC	29 ± 4	94 ± 16	303 ± 24	14 ± 3	16 ± 2
Solvent control (ethanol)	30 ±	97 ± 8	300 ± 6	16 ± 3	15 ± 1
15	-	-	309 ± 19	12 ± 4	-
50	33 ± 3	107 ± 10	288 ± 20	15 ± 3	15 ± 3
150	30 ± 5	94 ± 12	268 ± 13	12 ± 4	11 ± 4
500	27 ± 2	94 ± 14	195 ± 12	14 ± 4	13 ± 3
1500	31 ± 6	62 ± 5 (T)	210 ± 22	14 ± 1	9 ± 0
5000 (P)	29 ± 7	58 ± 8 (T)	161 ± 11 (T)	14 ± 2	4 ± 3 (T)
2-AA (0.8)	476 ± 85	545 ± 19	573 ± 26	116 ± 7	-
2-AA (1.7)	-	-	-	-	143 ± 19
NC = Negative Control P = precipitation of test compound T = bacteriotoxic 2-NF: 2-nitrofluorene; 9-AA: 9-aminoacridine; 2-AA: 2-aminoanthracene

Table 2: Test Results of Experiment 2

EXPERIMENT 2
S9-Mix	Without
Test item (µg/plate)	TA 98	TA 100	TA 102	TA 1535	TA 1537
NC	23 ± 7	113 ± 13	273 ± 13	12 ± 3	9 ± 2
Solvent control (ethanol)	23 ± 3	101 ± 6	293 ± 5	11 ± 3	9 ± 4
5	-	-	304 ± 17	-	-
15	-	103 ± 12	281 ± 10	-	8 ± 2
50	18 ± 3	97 ± 13	283 ± 15	15 ± 1	8 ± 3
150	16 ± 5	101 ± 18	270 ± 20	9 ± 2	8 ± 5
500	12 ± 6	90 ± 6	226 ± 28	9 ± 1	8 ± 2
1500	15 ± 3	104 ± 11	222 ± 18	9 ± 5	5 ± 2
5000 (P)	22 ± 3	71 ± 12 (T)	-	2 ± 1 (T)	-
NaN3 (0.7)	-	314 ± 6	-	292 ± 21	-
2-NF (2.5)	258 ± 17	-	-	-	-
9-AA (50)	-	-	-	-	848 ± 413
Mitomycin C (0.15)	-	-	987 ± 32	-	-
S9-Mix	With
Test item (µg/plate)	TA 98	TA 100	TA 102	TA 1535	TA 1537
NC	28 ± 2	96 ± 13	294 ± 7	7 ± 3	15 ± 5
Solvent control (ethanol)	27 ± 4	101 ± 5	290 ± 9	15 ± 3	15 ± 4
5	-	-	297 ± 17	-	12 ± 2
15	-	95 ± 12	249 ± 23	-	15 ± 5
50	25 ± 7	107 ± 7	220 ± 27	21 ± 4	11 ± 3
150	23 ± 3	114 ± 2	199 ± 16	19 ± 4	15 ± 6
500	28 ± 2	92 ± 10	205 ± 2	10 ± 3	11 ± 2
1500	22 ± 3	83 ± 16	156 ± 19 (T)	7 ± 3	10 ± 3
5000 (P)	19 ± 6	43 ± 12 (T)	-	6 ± 3 (T)	-
2-AA (0.8)	583 ± 26	839 ± 50	683 ± 27	123 ± 12	-
2-AA (1.7)	-	-	-	-	314 ± 29
NC = Negative Control P = precipitation of test compound T = bacteriotoxic 2-NF: 2-nitrofluorene; 9-AA: 9-aminoacridine; 2-AA: 2-aminoanthracene

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Under the experimental conditions described, the test substance was not mutagenic in the Ames test using the Salmonella typhimurium strains TA1535, TA1537, TA98, TA100 and TA102 in the presence and absence of a metabolic activation system.