Trideca-1,1,1,2,2,3,3,4,4,5,5,6,6-fluorohexane

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:

1994-04-06 t 1994-06-01

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1994

Report date:

1994

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals

Version / remarks:

Standards for Toxicity Investigations (Japan's Ministry of Labor, No. 77, September 1, 1988) and Notification on Partial Revision of Testing Methods Relating to the New Chemical Susbtances (Notification No. 700 of the Planning and Coordination Bureau, EA, No. 1039 of the Pharmaceutical Affairs Bureau, MHW & No. 1014 (1986) of the Basic Industries Bureau, MITI, December 5, 1986).

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:

Trideca-1,1,1,2,2,3,3,4,4,5,5,6,6-fluorohexane

EC Number:

206-581-9

EC Name:

Trideca-1,1,1,2,2,3,3,4,4,5,5,6,6-fluorohexane

Cas Number:

355-37-3

Molecular formula:

C6HF13

IUPAC Name:

1,1,1,2,2,3,3,4,4,5,5,6,6-tridecafluorohexane

Test material form:

liquid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Asahi Glass Co., Ltd.; Batch no. 31001
- Expiration date of the lot/batch: Not specified
- Purity test date: Not specified

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Stored in a cold and dark place
- Stability under test conditions: Not specified

FORM AS APPLIED IN THE TEST (if different from that of starting material): Colourless clear liquid

OTHER SPECIFICS:

Purity: 99.9%
Impurities: CClF2HCCl2F (0.05%)
Molecular Weight: 320.05
Boiling Point 71°C
Solubility: Oil soluble
Degree of Solublity Water (<10 mg/L)
DMSO (<5% w/v)
Acetone (≥ 10% w/v)

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

Rat Liver S9

Test concentrations with justification for top dose:

Dose range-finding test: 0, 50, 100, 200, 500, 1000, 2000, or 5000 µg/plate

Main Test: 0, 313, 625, 1250, 2500, or 5000 µg/plate (Based on the results of the dose range-finding test, 5000 µg/plate was selected as the highest dose and the lower four doses diluted with a geometric progression of 2 were set).

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: acetone
- Justification for choice of solvent/vehicle: Not specified

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

sodium azide

other: 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide (AF-2); 2-Aminoanthracene (2AA); 2-Methoxy-6-chloro--9-[3-(2-chloroethyl)-aminopropylamino]acridine.2HCl

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)
- Cell density at seeding (if applicable): Main Test - TA98: 2.0 x 10^9 viable cells/mL; TA100: 1.6 x 10^9 viable cells/mL; TA1535: 2.1 x 10^9 viable cells/mL; TA1537: 1.8 x 10^9 viable cells/mL; WP2 uvrA: 1.9 x 10^9 viable cells/mL

DURATION
- Preincubation period: 37 ± 0.5°C for 8-10 hours
- Expression time (cells in growth medium): 20 minutes
- Exposure duration: 48 hours

SELECTION AGENT (mutation assays): Not specified

NUMBER OF REPLICATIONS: 3 (negative control); 2 (Test substance and positive controls)

METHODS OF SLIDE PREPARATION AND STAINING TECHNIQUE USED: The appearance of revertant colonies (size and number), deposition of the test substance and growth inhibition were examined with a stereo microscope.

NUMBER OF CELLS EVALUATED: The number of revertant colonies was counted with a manual counter or a colony analyzer (CA-7, Toyo-sokki Co., Ltd.). The count using the colony analyzer was finalized after correcting with count drop. Each plate was measured three times, and the average of these three individual measurements with rounding off the figures below decimal point was adopted as the number of revertant colonies.

Evaluation criteria:

The test substance was judged to be positive, when the number of revertant colonies was increased more than twice that of the negative control in a dose-dependant manner, and the reproducibility of results was also obtained. It was judged to be negative in the other cases.

Statistics:

Statistical analysis was not applied.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

not examined

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

not examined

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

not examined

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

not examined

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

not examined

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

The sterility test confirmed the absence of micro-organisms.

Any other information on results incl. tables

Table 1. Results of the Main Test
With (+) or Without (-S9) Mix	Test Substance Dose (µg/plate)	Number of Revertants per Plate
		Base-pair Substitution Type			Frameshift Type
		TA 100	TA 1535	WP2uvrA	TA 98	TA 1537
-S9 Mix	Negative Control	131 153 (134) 117	12 8 (10) 9	44 47 (43) 37	22 20 (19) 14	10 12 (10) 8
	313	174 152 (163)	17 13 (15)	61 46 (54)	26 34 (30)	15 4 (10)
	625	119 113 (116)	9 18 (14)	52 53 (53)	17 24 (21)	22 5 (14)
	1250	162 158 (160)	14 15 (15)	51 43 (47)	24 28 (26)	5 11 (8)
	2500	147 147 (147)	10 17 (14)	48 42 (45)	19 29 (24)	11 12 (12)
	5000	147 147 (147)	11 15 (13)	41 43 (42)	26 16 (21)	8 7 (8)
+S9 Mix	Negative Control	148 148 (148) 148	21 28 (22) 16	51 47 (49) 49	34 29 (34) 39	24 19 (17) 8
	313	149 168 (159)	13 18 (16)	57 52 (55)	41 37 (39)	14 22 (18)
	625	133 143 (138)	11 14 (13)	50 61 (56)	43 48 (46)	22 13 (18)
	1250	147 134 (141)	14 12 (13)	53 52 (53)	29 30 (30)	11 25 (18)
	2500	112 147 (130)	12 14 (13)	55 41 (48)	38 30 (34)	17 19 (18)
	5000	158 138 (148)	15 22 (19)	54 56 (55)	36 35 (36)	15 20 (18)
Positive Control (-S9 Mix)	Chemical	AF-2	NaN3	AF-2	AF-2	ICR-191
	Dose (µg/plate)	0.01	0.5	0.01	0.1	1
	Number of Revertants /plate	424 398 (411)	295 297 (296)	444 412 (428)	395 445 (420)	1925 1948 (1937)
Positive Control (+S9 Mix)	Chemical	2AA	2AA	2AA	2AA	2AA
	Dose (µg/plate)	1	2	10	0.5	2
	Number of Revertants /plate	647 632 (640)	151 130 (141)	447 450 (449)	140 149 (145)	122 123 (123)

1) values in parenthesis show the mean of each plate

2) AF-2: 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide (AF-2)

3) 2AA: 2-Aminoanthracene

4) ICR-191: 2-Methoxy-6-chloro--9-[3-(2-chloroethyl)-aminopropylamino]acridine.2HCl

5) NaN₃: Sodium Azide

Applicant's summary and conclusion

Conclusions:

1,1,1,2,2,3,3,4,4,5,5,6,6-tridecafluorohexane was considered to have no reverse mutagenic potential under the conditions of the study.

Executive summary:

In a key bacterial reverse mutation assay, the in vitro mutagenic potential of the test material, 1,1,1,2,2,3,3,4,4,5,5,6,6-tridecafluorohexane, was evaluated in Salmonella typhimurium strains TA98, TA100, T1535, and TA 1537 and Escherischia coli strain WP2uvrA using pre-incubation method in the absence and presence of a metabolic activation system (±S9).

 

Based on a dose range-finding study, the concentrations of the test material used in the main study were 0, 313, 625, 1250, 2500, or 5000 µg/plate.

 

It was observed that the number of revertant colonies in all strains was less than twice that of the negative control at all dose levels tested in the absence and presence of metabolic activation (±S9). The positive controls exhibited a significant increase in the number of revertant colonies and the number of revertants for positive as well as negative controls was within the historical range.

 

Based on the results observed in the study, 1,1,1,2,2,3,3,4,4,5,5,6,6-tridecafluorohexane was considered to be negative in this bacterial reverse mutation assay.