4-pentylcyclohexanone

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:

May 16, to July 13, 1990

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1990

Report date:

1990

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

1981

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

4-pentylcyclohexanone

EC Number:

406-670-4

EC Name:

4-pentylcyclohexanone

Cas Number:

61203-83-6

Molecular formula:

C11H20O

IUPAC Name:

4-pentylcyclohexan-1-one

Method

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:

S. typhimurium TA 1538

Metabolic activation:

with and without

Metabolic activation system:

Due to migration, the value was transferred to one of the current document's attachments

Test concentrations with justification for top dose:

1st series of experiments:
5, 10, 50, 250, 500, and 750 µg/plate
2nd and additional series of experiments:
1, 5, 10, 50, 250, and 375 µg/plate (TA 98, TA 1535, TA 1537, and TA 1538)
1, 5, 10, 50, 250, and 500 µg/plate (TA 100)

The test material concentrations used were selected according to the OECD Guideline for this test system. They were chosen to cover a broad concentration range including, if possible, also a toxic concentration.

Vehicle / solvent:

- Vehicle/solvent used: The test material was dissolved in dimethylsulfoxide (DMSO).

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

without S9 mix

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

without S9 mix

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 4-Nitro-1,2-phenylene diamine

Remarks:

without S9 mix

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

without S9 mix

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

without S9 mix

Controls 6

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: Daunomycin

Remarks:

without S9 mix

Controls 7

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracene

Remarks:

with S9 mix

Details on test system and experimental conditions:

NUMBER OF REPLICATIONS:
- Number of cultures per concentration: 4
- Number of independent experiments : 3

METHOD OF TREATMENT/ EXPOSURE:
- Test substance added in agar (plate incorporation)

TREATMENT AND HARVEST SCHEDULE:
- Exposure duration/duration of treatment: 48 h

METHODS FOR MEASUREMENT OF CYTOTOXICITY
- Method: reduction in the number of spontaneous revertants, clearing of the background lawn, reduced survival of treated cultures

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 500 µg/plate with and without S9 mix

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 500 µg/plate with and without S9 mix

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

from 500 µg/plate onwards with and without S9 mix

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 500 µg/plate with and without S9 mix

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 375 µg/plate with and without S9 mix

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES: In a range finding test (using S.typh. TA 100 and TA 1535 without metabolic activation) toxicity was investigated in a wide range of test material concentrations (50 - 10000 µg/plate). Toxicity normally becomes evident by a reduction in the number of spontaneous revertants, a clearing of the background lawn, or by reduced survival of treated cultures. These effects were observed with the test item in the range finding test at concentrations > 500 µg/plate.
Thus, in the first series of experiments the following concentrations were tested:
5, 10, 50, 250, 500, and 750 µg/plate.
As for the highest concentrations tested strong cytotoxic effects still were observed during the first series of experiments, the second and additional series of experiments were performed using the following concentrations:
TA 98, TA 1535, TA 1537, and TA 1538: 1, 5, 10, 50, 250, and 375 µg/plate
TA 100: 1, 5, 10, 50, 250, and 500 µg/plate

- Signs of toxicity
Because of cytotoxic effects only four concentrations were valid in the first series of experiments (except TA 100). As, however, five concentrations are recommended in the OECD- and EEC- Guidelines, an additional series of experiments was performed, using the same concentrations as in the second series.

- Individual plate counts
see table 1-4
- Mean number of revertant colonies per plate and standard deviation
see table 1-4

Any other information on results incl. tables

Table 1: Results of second series (TA100)

Without S-9 Mix

Conc.(µg/pl)		Individual Values				n	Mean	SD	Pc	Bg
	0.0	124	127	140	144	4	134	9.7	-	-
	1.0	113	144	135	137	4	132	13.4	-	-
	5.0	123	123	120	125	4	123	2.1	-	-
	10.0	139	148	154	149	4	148	6.2	-	-
	50.0	153	162	152	163	4	158	5.8	-	-
	250.0	118	120	108	117	4	116	5.3	-	-
	500.0	55	74	70	59	4	65	9.0	-	+
	0.0	153	149	140	152	4	149	5.9	-	-
2-AA	1.0	150	140	138	163	4	148	11.4	-	-
MMS	500.0	1276	1295	1202	1128	4	1225	76.2	-	-
ENNG	4.0	906	866	721	855	4	837	80.4	-	-

 

With S-9 Mix

Conc.(µg/pl)		Individual Values				n	Mean	SD	Pc	Bg
	0.0	192	164	192	213	4	190	20.1	-	-
	1.0	170	174	189	166	4	175	10.0	-	-
	5.0	189	226	200	208	4	206	15.6	-	-
	10.0	190	166	162	178	4	174	12.6	-	-
	50.0	165	194	217	162	4	185	26.0	-	-
	250.0	161	153	175	177	4	167	11.5	-	-
	500.0	0	0	0	0	4	0	0.0	-	+
	0.0	201	190	166	179	4	184	15.0	-	-
2-AA	1.0	541	624	528	637	4	583	55.9	-	-

Table 2: Results of second series (TA1535)

Without S-9 Mix

Conc.(µg/pl)		Individual Values				n	Mean	SD	Pc	Bg
	0.0	7	16	13	19	4	14	5.1	-	-
	1.0	19	9	16	8	4	13	5.4	-	-
	5.0	26	21	18	13	4	20	5.4	-	-
	10.0	20	20	18	20	4	20	1.0	-	-
	50.0	16	28	8	7	4	15	9.7	-	-
	250.0	7	9	6	17	4	10	5.0	-	-
	375.0	2	9	8	1	4	5	4.1	-	+
	0.0	19	16	16	15	4	17	1.7	-	-
2-AA	1.0	13	17	21	27	4	20	6.0	-	-
ENNG	10.0	863	815	814	798	4	823	28.1	-	-
Ve-H2O		17	17	17	18	4	17	0.5	-	-
NaN3	1.0	448	479	463	378	4	442	44.5	-	-

 

 

With S-9 Mix

Conc.(µg/pl)		Individual Values				n	Mean	SD	Pc	Bg
	0.0	15	14	13	16	4	15	1.3	-	-
	1.0	21	18	20	17	4	19	1.8	-	-
	5.0	14	18	27	22	4	20	5.6	-	-
	10.0	15	14	15	20	4	16	2.7	-	-
	50.0	20	9	14	13	4	14	4.5	-	-
	250.0	19	16	15	16	4	17	1.7	-	-
	375.0	9	7	5	7	4	7	1.6	-	+
	0.0	21	7	26	15	4	17	8.2	-	-
2-AA	1.0	141	122	118	137	4	130	11.2	-	-

 

 

Table 3: Results of second series (TA1537)

Without S-9 Mix

Conc.(µg/pl)		Individual Values				n	Mean	SD	Pc	Bg
	0.0	5	3	4	6	4	5	1.3	-	-
	1.0	3	3	4	2	4	3	0.8	-	-
	5.0	5	6	5	2	4	5	1.7	-	-
	10.0	3	1	3	2	4	2	1.0	-	-
	50.0	6	7	7	10	4	8	1.7	-	-
	250.0	6	2	4	4	4	4	1.6	-	-
	375.0	1	0	1	2	4	1	0.8	-	+
	0.0	4	6	6	4	4	5	1.2	-	-
2-AA	1.0	5	7	8	5	4	6	1.5	-	-
EtOH		4	3	3	4	4	4	0.6	-	-
9-AA	20	30	35	43	42	4	38	6.1	-	-
9-AA	50	556	574	426	517	4	518	65.9	-	-

 

With S-9 Mix

Conc.(µg/pl)		Individual Values				n	Mean	SD	Pc	Bg
	0.0	5	3	6	6	4	5	1.4	-	-
	1.0	6	4	5	14	4	7	4.6	-	-
	5.0	5	4	4	5	4	5	0.6	-	-
	10.0	7	13	3	2	4	6	5.0	-	-
	50.0	2	5	14	5	4	7	5.2	-	-
	250.0	4	6	3	6	4	5	1.5	-	-
	375.0	2	6	1	4	4	3	2.2	-	+
	0.0	9	4	5	4	4	6	2.4	-	-
2-AA	1.0	41	55	61	44	4	50	9.4	-	-

Table 4: Results of second series (TA1538)

Without S-9 Mix

Conc.(µg/pl)		Individual Values				n	Mean	SD	Pc	Bg
	0.0	5	2	9	4	4	5	2.9	-	-
	1.0	8	6	3	7	4	6	2.2	-	-
	5.0	5	3	9	3	4	5	2.8	-	-
	10.0	2	4	4	8	4	5	2.5	-	-
	50.0	6	6	4	2	4	5	1.9	-	-
	250.0	3	7	6	3	4	5	2.1	-	-
	375.0	0	0	1	2	4	1	1.0	-	+
	0.0	6	1	5	5	4	4	2.2	-	-
2-AA	1.0	6	7	8	6	4	7	1.0	-	-
4-NP	10.0	1299	1320	1205	1177	4	1250	69.9	-	-
2-NF	5.0	783	741	543	388	4	614	183.3	-	-

 

 

With S-9 Mix

Conc.(µg/pl)		Individual Values				n	Mean	SD	Pc	Bg
	0.0	14	14	21	8	4	14	5.3	-	-
	1.0	7	14	7	4	4	8	4.2	-	-
	5.0	9	9	16	5	4	10	4.6	-	-
	10.0	c	c	c	c	0	-	-	-	-
	50.0	5	9	8	5	4	7	2.1	-	-
	250.0	6	5	7	8	4	7	1.3	-	-
	375.0	1	1	8	4	4	4	3.3	-	+
	0.0	15	18	14	15	4	16	1.7	-	-
2-AA	1.0	287	315	310	274	4	297	19.3	-	-

 

Applicant's summary and conclusion

Conclusions:

Under the test conditions used, the test item was not mutagenic in Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537, and TA 1538 with or without addition of S9 mix.

Executive summary:

The investigations for mutagenic potential were performed according OECD TG 471 using Salmonella typhimurium TA 100, TA 98, TA 1535, TA 1537, and TA 1538 as tester strains. The plate incorporation test with and without addition of rat liver S-9 fraction (Aroclor-induced) was used.

The substance was tested in three series of experiments at the following concentrations:

First series of experiments: 5, 10, 50, 250, 500, and 750 µg/plate

Second and additional series of experiments: 

1, 5, 10, 50, 250, and 375 µg/plate (TA 98, TA 1535, TA 1537, and TA 1538)

1, 5, 10, 50, 250, and 500 µg/plate (TA 100)

9-Aminoacridine, daunomycin, 1-ethyl-2-nitro-3-nitrosoguanidine, methyl methanesulfonate, 2-nitrofluorene, 4-nitro-1,2-phenylene diamine, and sodium azide served as positive control compounds for testing the bacteria. 2-Aminoanthracene was used for testing the bacteria and the activity of the S-9 preparation.

The positive controls showed normal reversion properties of all strains and good metabolic activity of the S-9 mix used.

With and without addition of S-9 as the metabolizing system, the substance did not show any mutagenic activity in the concentration range used.