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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP Guidance study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Qualifier:
according to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Reference substance name:
1,2,3-Benzotriazole
IUPAC Name:
1,2,3-Benzotriazole
Constituent 2
Chemical structure
Reference substance name:
Benzotriazole
EC Number:
202-394-1
EC Name:
Benzotriazole
Cas Number:
95-14-7
Molecular formula:
C6H5N3
IUPAC Name:
benzotriazole
Test material form:
other: needles
Details on test material:
- Name of test material (as cited in study report): 1,2,3-Benzotriazole
- Physical state: solid
- Analytical purity: 99.87 %
- Lot/batch No.: 111254 / 112649
- Expiration date of the lot/batch: Nov. 2012
- Storage condition of test material: 20 +- 5°C

Method

Target gene:
his-
Species / strain
Species / strain / cell type:
S. typhimurium, other: TA 97a, TA 98, TA 100, TA 102, TA1535
Metabolic activation:
with and without
Test concentrations with justification for top dose:
first experiment
50/150/500/1501/5003 µg/plate
second experiment
313/625/1250/2500/5000 µg/plate
Vehicle / solvent:
Dimethylsulfoxide (DMSO)

DMSO is chosen because 1H-benzotriazole is completly soluble and the vehicle does not have any effects on the viability of the bacetria.
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
sodium azide
benzo(a)pyrene
other: 4-Nitro-1,2-phenylenediamine; 2-Amino-anthracene
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation) with and without preincubation

DURATION
- Preincubation period: 20 min

NUMBER OF REPLICATIONS: 4
Evaluation criteria:
The colonies were counted visually.
The increase factor f(I) of revertant induction and the absolute number ov refertants were calculated.
If a significant, reproducible increase of revertant colonies (f(I) > 2) can be observed, 1H-benzotriazole is considered mutagenic.
Statistics:
mean values and standard deviations of each treatment, solvent control and positive control are calculated. Calculations were performed with unrounded values

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 97a
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
COMPARISON WITH HISTORICAL CONTROL DATA:
The negative, vehicle and positive controls were compared to historical data and
found to be in the range of this data.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table: Mean revertants first experiment

 Strain        TA97a TA98         TA100 TA102 TA1535
 Induction  -S9  +S9 -S9   +S9  -S9  +S9  -S9  +S9  -S9 +S9 
 H2O Mean   118 121  15  19  90  101  145  144  16  17 
   sd  8 6  3  2  9  9  9  20  3  2 
 DMSO Mean   126 121  19  20  97  94  127  140  17  15 
   sd  9 7  3  3  12  14  9  8  2  3 
 Positive controls  Mean 563  562  236  220  452  470  583  631  193  186 
   sd  69 37  34  12  41  26  102  66  13  14 
   f(I)  4.47 4.64  12.42  11.00  5.02  5.00  4.59  4.51  12.06  12.40 
 5003 µg/pl.  Mean  107 125  18  15  99  102  149  141   16 14 
   sd  5 4  2  4  18  6  10  9  5  1 
   f(I)  0.85 1.03  0.95  0.75  1.02  1.09  1.17  1.01  0.94  0.93 
 1501 µg/pl.  Mean  123 124  17  16  103  103  143  146  17  17 
   sd  10 5  2  4  11  7  7  7  3  2 
   f(I)  0.98 1.02  0.89  0.80  1.06  1.10  1.13  1.04  1.00  1.13 
 500 µg/pl.  Mean  120 118  15  16  89  110  139  130  15  14 
   sd  5 10  5  1  8  3  16  11  4  1 
   f(I)  0.95 0.98  0.79  0.80  0.92  1.17  1.09  0.93  0.88  0.93 
 150 µg/pl.  Mean  116 121  14  18  88  101  136  141  15  15 
   sd  3 7  1  3  21  10  5  11  2  1 
   f(I)  0.92 1.00  0.74  0.90  0.91  1.07  1.07  1.01  0.88  1.00 
 50 µg/pl.  Mean  119 108  16  20  83  97  137  128  15  15 
   sd  5 7  3  3  19  6  4  15  2  2 
   f(I)  0.94 0.89  0.84  1.00  0.86  1.03  1.08  0.91  0.88  1.00 

Table: Mean revertants second experiment

 Strain        TA97a TA98         TA100 TA102 TA1535
 Induction  -S9  +S9 -S9   +S9  -S9  +S9  -S9  +S9  -S9 +S9 
 H2O Mean   118 117  15  15  100  87  166  153  16  15 
   sd  4 3  3  1  8  10  14  11  2  2 
 DMSO Mean   117 115  18  16  98  96  149  154  18  16 
   sd  3 6  3  1  3  3  23  7  3  2 
 Positive controls  Mean  517 592  217  222  416  473  621  611  210  241 
   sd  71 13  12  15  38  26  25  43  10  13 
   f(I)  4.42 5.15  12.06  13.88  4.16  4.93 4.17 3.97  13.13 15.06 
 5002 µg/pl.  Mean 104  112  15  14  95  93  139  146  15  14 
   sd  7 7  2  3  7  7  12  5  2  3 
   f(I)  0.89 0.97  0.83  0.88  0.97  0.97  0.93  0.95  0.83  0.88 
 2501 µg/pl.  Mean 112  113  14  15  84  79  145  157  15  13 
   sd  3 3  3  3  8  9  5  16  2  1 
   f(I)  0.96 0.98  0.78  0.94  0.86  0.82  0.97  1.02  0.83  0.81 
 1251 µg/pl.  Mean 117  109  14  16  82  79  143  150  17  16 
   sd  4 6  2  2  8  6  11  16  4  4 
   f(I)  1.00 0.95  0.78  1.00  0.84  0.82  0.96  0.97  0.94  1.00 
 626 µg/pl.  Mean 116  116  15  17  92  87  156  157  15  15 
   sd  4 4  1  3  16  9  14  12  2  3 
   f(I)  0.99 1.01  0.83  1.06  0.94  0.91  1.05  1.02  0.83  0.94 
 313 µg/pl.  Mean 115  114  14  17  85  85  146  151  16  14 
   sd  2 2  2  3  11  7  5  8  2  2 
   f(I)  0.98 0.99  0.78  1.06  0.87  0.89  0.98  0.98  0.89  0.88 

Applicant's summary and conclusion

Conclusions:
1H-benzotriazole did not show mutagenic effects in the two conducted experiments.