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Toxicological information

Genetic toxicity: in vitro

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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:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
not specified
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
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:
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:
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 + 9.0c  122 + 2.4  99 + 3.0  135 + 2.9c  102 + 8.7  138 + 3.8
 10,000  65 + 3.7c  104 + 4.4c  103 + 13.5c  109 + 6.4c  91 + 7.8  113 + 7.2c
 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 + 3.0c  27 + 2.7  9 + 0.9  12 + 3.5  9 + 3.8  13 + 1.2
 10,000  22 + 4.8c  21 + 1.5c  9 + 1.5c  12 + 0.6  11 + 1.5c  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 + 2.1c  128 + 2.9  130 + 7.1  152 + 6.0c  129 + 2.2  167 + 6.4
 10,000  92 + 5.2c  100 + 5.6c  131 + 2.2c  172 + 7.8c  110 + 4.6c  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 + 1.2c  16 + 1.5  33 + 5.5  30 + 3.5  38 + 2.2  35 + 2.2
 10,000  14 + 1.0c  19 + 3.5c  34 + 1.2  28 + 1.8  31 + 3.8  36 + 1.7c
 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.