Benzyltrimethylammonium chloride

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:

No dates listed in report

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to guideline and/or standard method but was non-GLP.

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

2000

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

not specified

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Benzyltrimethylammonium chloride

EC Number:

200-300-3

EC Name:

Benzyltrimethylammonium chloride

Cas Number:

56-93-9

Molecular formula:

C10H16N.Cl

IUPAC Name:

benzyltrimethylazanium chloride

Details on test material:

Benzyltrimethylammonium chloride was obtained from Fluka Chemical Corporation (Ronkonkoma, NY) in one lot (306793/1). Information on the identity, purity, and stability of the bulk chemical was provided by the manufacturer; identity was confirmed by the study laboratory. Reports on analyses performed in support of the benzyltrimethylammonium chloride studies are on file at the National Institute of Environmental Health Sciences.

The manufacturer identified the chemical, an off-white to yellow crystalline powder, as benzyltrimethylammonium chloride by nuclear magnetic resonance spectroscopy. The purity of lot 306793/1, determined by argentometric titration, was 100.4% or greater. The study laboratory confirmed the identity of the chemical with infrared spectroscopy. The spectrum was consistent with a literature reference for benzyltrimethylammonium bromide (Aldrich, 1990).

Based on the manufacturers stability information, the bulk chemical was stored at room temperature in sealed containers flushed with nitrogen to expel moisture.

Reference
Aldrich Catalog/Handbook of Fine Chemicals 1990-1991 (1990), p. 146. Aldrich Chemical Company, Inc., Milwaukee, WI.

Method

Target gene:

Histidiine dependent strain of bacteria used to detect point mutations.

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA97, TA98, TA100, and TA1535

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254-induced male Sprague-Dawley rat or Syrian hamster liver

Test concentrations with justification for top dose:

0, 100, 333, 1000, 3333 and 10,000 ug/plate

Vehicle / solvent:

Not specified in report, probably water.

Details on test system and experimental conditions:

Benzyltrimethylammonium chloride was sent to the laboratory as a coded aliquot from Radian Corporation (Austin, TX). It was incubated with the Salmonella typhimurium tester strains TA97, TA98, TA100, and TA1535, either in buffer or S9 mix (metabolic activation enzymes and cofactors from Aroclor 1254-induced male Sprague-Dawley rat or Syrian hamster liver) for 20 minutes at 37 C. Top agar supplemented with L-histidine and d-biotin was added, and the contents of the tubes were mixed and poured onto the surfaces of minimal glucose agar plates. Histidine-independent mutant
colonies arising on these plates were counted following incubation for 2 days at 37 C.

Each trial consisted of triplicate plates of concurrent positive and negative controls and five doses of benzyltrimethylammonium chloride. In the absence of toxicity, 10,000 µg/plate was selected as the high dose.

Evaluation criteria:

In this assay, a positive response is defined as a reproducible, dose-related increase in histidine-independent (revertant) colonies in any one strain/activation combination. An equivocal response is defined as an increase in revertants that is not dose related, is not reproducible, or is not of sufficient magnitude to support a determination of mutagenicity. A negative response is obtained when no increase in revertant colonies is observed following chemical treatment. There is no minimum percentage or fold increase required for a chemical to be judged positive or weakly positive.

Statistics:

Means and Standard Deviations were calculated.

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA97, TA98, TA100, or TA1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

At the two highest doses tested.

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

Benzyltrimethylammonium chloride (100 to 10,000 µg/plate) was not mutagenic in Salmonella typhimurium strain TA97, TA98, TA100, or TA1535 with or without induced rat or hamster liver S9 metabolic activation enzymes (Table 1). Slight toxicity was noted at the two highest concentrations tested in all four strains.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1 Mutagenicity of Benzyltrimethylammonium Chloride in Salmonella typhimurium

	Revertants/plate ^b
Dose	-S9		+ hamster S9		+ rat S9
(µg/plate)	Trial 1	Trial 2	Trial 1	Trial 2	Trial 1	Trial 2
Strain TA100
0	122 + 8.2	122 + 9.2	116 + 7.0	119 + 11.0	84 + 6.6	147 + 9.6
100	119 + 12.7	128 + 3.7	102 + 1.2	130 + 4.9	105 + 7.1	128 + 7.1
333	115 + 6.9	118 + 14.2	110 + 3.5	115 + 9.0	89 + 6.9	128 + 3.4
1000	116 + 11.7	116 + 8.2	107 + 8.1	113 + 5.7	104 + 8.7	136 + 2.5
3333	121 + 9.0^c	122 + 2.4	99 + 3.0	135 + 2.9^c	102 + 8.7	138 + 3.8
10,000	65 + 3.7^c	104 + 4.4^c	103 + 13.5^c	109 + 6.4^c	91 + 7.8	113 + 7.2^c
Positive controld	1389 + 22.6	1485 + 4.8	1125 + 21.5	2231 + 35.7	1883 + 33.0	702 + 16.7
Strain TA1535
0	31 + 3.0	29 + 0.9	9 + 0.9	14 + 0.9	9 + 1.7	12 + 2.3
100	29 + 0.7	30 + 1.0	11 + 3.8	8 + 1.0	10 + 1.9	13 + 0.0
333	25 + 0.9	26 + 4.4	13 + 2.3	12 + 1.5	10 + 1.5	15 + 2.0
1000	28 + 1.5	37 + 2.2	9 + 1.2	10 + 1.7	11 + 0.7	11 + 0.7
3333	28 + 3.0^c	27 + 2.7	9 + 0.9	12 + 3.5	9 + 3.8	13 + 1.2
10,000	22 + 4.8^c	21 + 1.5^c	9 + 1.5^c	12 + 0.6	11 + 1.5^c	11 + 2.3
Positive control	1051 + 8.7	1095 + 18.9	127 + 9.0	323 + 17.8	117 + 5.0	130 + 13.9
Strain TA97
0	104 + 8.4	121 + 3.5	140 + 4.1	170 + 8.8	113 + 9.7	181 + 5.5
100	113 + 7.0	110 + 7.4	143 + 0.3	158 + 8.8	126 + 12.1	181 + 4.2
333	99 + 0.9	114 + 3.6	144 + 8.0	156 + 1.9	124 + 3.5	155 + 9.2
1000	112 + 8.3	128 + 5.5	139 + 6.9	165 + 6.1	133 + 5.3	168 + 11.1
3333	95 + 2.1^c	128 + 2.9	130 + 7.1	152 + 6.0^c	129 + 2.2	167 + 6.4
10,000	92 + 5.2^c	100 + 5.6^c	131 + 2.2^c	172 + 7.8^c	110 + 4.6^c	174 + 12.8
Positive control	1626 + 161.0	722 + 9.3	511 + 17.2	1452 + 23.1	763 + 36.3	691 + 13.3
Strain TA98
0	19 + 4.1	16 + 0.6	39 + 2.6	35 + 2.7	32 + 5.5	31 + 5.8
100	14 + 1.5	14 + 1.7	37 + 3.4	27 + 4.4	31 + 1.2	29 + 0.9
333	16 + 2.1	19 + 4.1	32 + 3.2	31 + 2.8	30 + 1.3	34 + 2.6
1000	17 + 2.0	17 + 2.4	36 + 2.1	34 + 1.2	35 + 1.7	30 + 5.8
3333	19 + 1.2^c	16 + 1.5	33 + 5.5	30 + 3.5	38 + 2.2	35 + 2.2
10,000	14 + 1.0^c	19 + 3.5^c	34 + 1.2	28 + 1.8	31 + 3.8	36 + 1.7^c
Positive control	1246 + 69.5	1848 + 57.8	1205 + 26.9	2210 + 28.6	1401 + 30.6	659 + 32.9

^b Revertants are presented as mean ± standard error from three plates.

^c Slight toxicity

^d The positive controls in the absence of metabolic activation were sodium azide (TA100 and TA1535), 9-aminoacridine (TA97), and 4-nitro-o-phenylenediamine (TA98). The positive control for metabolic activation with all strains was 2-aminoanthracene.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Benzyltrimethylammonium chloride was not mutagenic in Salmonella typhimurium strains TA97, TA98, TA100, and TA1535, with or without S9 metabolic activation enzymes.

Executive summary:

Benzyltrimethylammonium chloride was not mutagenic in Salmonella typhimurium strains TA97, TA98, TA100, and TA1535, with or without S9 metabolic activation enzymes.