1-Propanaminium, 3-amino-N-(carboxymethyl)-N,N-dimethyl-, N-coco acyl derivs., hydroxides, inner salts

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data is from publication

Data source

Reference

Reference Type:

publication

Title:

Gene mutation toxicity study of test chemical

Author:

Hidesuke SHIMIZU et. al.

Year:

1985

Bibliographic source:

Jpn. J. Ind. Health.,1985

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

To determine the gene mutation toxicity of test chemical.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

N,N'-ethylenedi(stearamide)

EC Number:

203-755-6

EC Name:

N,N'-ethylenedi(stearamide)

Cas Number:

110-30-5

Molecular formula:

C38H76N2O2

IUPAC Name:

N,N'-ethylenedi(stearamide)

Details on test material:

IUPAC name: N,N'-ethylenedi(stearamide)
Mol. formula: C38H76N2O2
Molecular Weight : 593.03 gm/mol
Smiles: CCCCCCCCCCCCCCCCCC(=O)NCCNC(=O)CCCCCCCCCCCCCCCCC
InChI: InChI=1S/C38H76N2O2/c1-3-5-7-9-11-13-15-17-19-21-23-25-27-29-31-33-37(41)39-35-36-40-38(42)34-32-30-28-26-24-22-20-18-16-14-12-10-8-6-4-2/h3-36H2,1-2H3,(H,39,41)(H,40,42)

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA 1535, TA 1537, TA 98, TA 100, TA 1538 and E. coli WP2

Metabolic activation:

with and without

Metabolic activation system:

Type and composition of metabolic activation system:
- source of S9 : liver of male Sprague-Dawley rats
- method of preparation of S9 mix : The S9 fraction was prepared from the liver of male Sprague-Dawley rats, weighing about 220g (seven weeks old), pretreated with polychlorinated biphenyl (KC 500) at a dose of 500mg/kg body weight five days before sacrifice.
- concentration or volume of S9 mix and S9 in the final culture medium : 0.1 ml
- quality controls of S9 (e.g., enzymatic activity, sterility, metabolic capability)

Test concentrations with justification for top dose:

1, 5, 10, 50, 100, 500, 1000 and 5000 µg/plate

Vehicle / solvent:

Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test chemical was soluble in DMSO

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

9-aminoacridine

2-nitrofluorene

N-ethyl-N-nitro-N-nitrosoguanidine

benzo(a)pyrene

other: 2-(2-Furyl)-3-(5-nitro-2-furyl) acrylamide (AF-2) and 9-aminoacridine

Details on test system and experimental conditions:

NUMBER OF REPLICATIONS:
- Number of cultures per concentration: duplicate
- Number of independent experiments : not specified

METHOD OF TREATMENT/ EXPOSURE:
- Cell density at seeding (if applicable): not specified
- Test substance added in medium; in agar (plate incorporation): 0.1ml of bacterial strain and 0.5ml of S9 mix or sodium phosphate buffer (pH 7.4) were added to a sterile test tube containing 0.1ml of various concentrations of the chemicals. This mixture was preincubated in a shaker water bath at 37°C for 20 minutes, and then added to 2ml of molten top agar (45°C).

TREATMENT AND HARVEST SCHEDULE:
- Preincubation period, if applicable: 20 minutes
- Exposure duration/duration of treatment: 48 hours
- Harvest time after the end of treatment (sampling/recovery times): not specified

Rationale for test conditions:

not specified

Evaluation criteria:

Number for revertants analysed

Statistics:

not specified

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA 1535, TA 1537, TA 98, TA 100, TA 1538 and E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Remarks:

DMSO

Untreated negative controls validity:

not specified

True negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

Not specified

Remarks on result:

other: No mutagenic potential observed

Any other information on results incl. tables

Table 1: Mutagenic activity of test chemical

Compound	Dose µg/ plate	Revertant colonies/ plate
		TA 100		TA 1535		WP2 uvrA		TA98		TA 1537		TA 1538
		-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
DMSO (Control)		150±16.8	154±17.5	30±5.6	15±7.1	30±9.7	34±10.9	32±7.7	42±10.1	18±8.6	22±6.3	22±7.3	28±5.7
Positive control
AF-2	0.01	501±84.7	-b	-	-	-	-	-	-	-	-	-	-
	0.05	-	-	-	-	1082±293.7	-	278±64.8	-	-	-	-	-
ENNG	0.5	-	-	1101±683.1	-	-	-	-	-	-	-	-	-
9AC	80.0	-	-	-	-	-	-	-	-	889±275.7	-	-	-
4NQO	0.25	-	-	-	-	-	-	-	-	-	-	270±66.2	-
B(a)P	5.0	-	1084±236.3	-	-	-	-	-	809±108.4	-	313±41.6	-	354±89.4
2AA	5.0	-		-	440±198.6	-	359±127.0	-	-	-	-	-	-
Test chemical	1	150	169	35	19	22	53	28	40	6	12	16	28
	5	156	162	29	20	33	42	27	36	13	13	23	22
	10	141	166	35	11	39	36	31	35	5	11	21	34
	50	152	161	32	13	31	33	23	35	8	12	19	23
	100	165	161	39	14	31	39	23	31	12	8	20	20
	500	152	163	26	25	22	34	31	37	10	13	24	30
	1000	176	168	31	20	28	39	34	56	10	15	18	27
	5000	139	185	24	23	25	41	19	28	8	10	17	16

-b: Not tested

Applicant's summary and conclusion

Conclusions:

Test chemical did not induce mutation in Salmonella Typhimurium TA 1538, 1535, 1537, 98, 100 and E. coli WP2 and hence it is not likely to classify as a gene mutant in vitro.

Executive summary:

Gene mutation study was performed on Salmonella Typhimurium strains to determine the mutagenic nature of test chemical. Salmonella Typhimurium TA 1538, 1535, 1537, 98, 100 and E. coli WP2 were used during the study. The study was performed in the presence and absence of S9 metabolic activation. The S9 fraction was prepared from the liver of male Sprague-Dawley rats, weighing about 220g (seven weeks old), pretreated with polychlorinated biphenyl (KC 500) at a dose of 500mg/kg body weight five days before sacrifice. The positive controls used were 4-nitroquinoline-N-oxide, 9-aminoacridine, 2-nitrofluorene, N-ethyl-N-nitro-N-nitrosoguanidine, benzo(a)pyrene, 2-(2-Furyl)-3-(5-nitro-2-furyl) acrylamide (AF-2) and 9-aminoacridine. 0.1ml of bacterial strain and 0.5ml of S9 mix or sodium phosphate buffer (pH 7.4) were added to a sterile test tube containing 0.1ml of various concentrations of the chemical. The test concentrations used were 1, 5, 10, 50, 100, 500, 1000 and 5000 µg/plate and the test chemical was dissolved in DMSO. This mixture was preincubated in a shaker water bath at 37°C for 20 minutes, and then added to 2ml of molten top agar (45°C). This mixture was preincubated in a shaker water bath at 37°C for 20 minutes, and then added to 2ml of molten top agar (45°C). The contents of each tube were mixed and poured onto a minimal glucose agar plate immediately. The plates were incubated at 37°C for 48 hours, and then the number of revertant colonies on each plate was scored with an automated colony counter. The background bacterial lawn was checked routinely by dissected microscope. Negleble amount of revertant colonies were observed as compared to positive control substances. Thus the test chemical can be considered as non-mutagenic when Salmonella Typhimurium TA 1538, 1535, 1537, 98, 100 and E. coli WP2 were treated in the presence and absence of S9 activation system.