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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:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2019
Report date:
2020

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Sodium (xylenes and 4-ethylbenzene) sulfonates
EC Number:
701-037-1
Cas Number:
1300-72-7
Molecular formula:
-
IUPAC Name:
Sodium (xylenes and 4-ethylbenzene) sulfonates

Method

Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Remarks:
Permanent stocks of these strains are kept at -80°C in ERBC facilities.
Metabolic activation:
with and without
Metabolic activation system:
Two batches of S9 tissue fraction, provided by Trinova Biochem GmbH, were used in this study and had the following characteristics:
Species Rat
Strain Sprague Dawley
Tissue Liver
Inducing Agents Phenobarbital – 5,6-Benzoflavone
Producer MOLTOX,Molecular Toxicology, Inc.
Batch Number 4086 (Main Assays)

The mixture of S9 tissue fraction and cofactors (S9 mix) was prepared as follows (for each 10 mL):
S9 tissue fraction 1.0mL
NADP (100 mM) 0.4mL
G-6-P (100 mM) 0.5mL
KCl (330 mM) 1.0mL
MgCl2 (100 mM) 0.8mL
Phosphate buffer (pH 7.4, 200 mM) 5.0mL
DistilledWater 1.3mL
Test concentrations with justification for top dose:
Toxicity test: 5000, 1580, 500, 158 and 50 µg/plate
I and II Main assay: 5000, 2500, 1250, 625 and 313 µg/plate (no effect obsterved in the first main test and the same doses were used for the second main test)
Vehicle / solvent:
Water for injection
Controlsopen allclose all
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
TA1537 without metabolic activation
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
TA1535 and TA100 wihtout metabolic activation
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
2-nitrofluorene
Remarks:
TA98 without metabolic activation
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
methylmethanesulfonate
Remarks:
E.Coli WP2 uvrA wihtout metabolic activation
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene
Remarks:
All strains with metabolic activation
Details on test system and experimental conditions:
Solubility of the test item was evaluated in a preliminary trial using sterile water for injection. This solvent was selected since it is compatible with the survival of the bacteria and the S9 metabolic activity.

Two Main Assays were performed including negative and positive controls in the absence and presence of an S9 metabolising system. Three replicate plates were used at each test point.
The Main Assay I was performed using a plate-incorporation method. The components of the assay (the tester strain bacteria, the test item and S9 mix or phosphate buffer) were added to molten overlay agar and vortexed. The mixture was then poured onto the surface of a minimal medium agar plate and allowed to solidify prior to incubation.

The overlay mixture was composed as follows:
Overlay agar (held at 45°C) 2.0mL
Test or control item solution 0.1mL
S9 mix or phosphate buffer (pH 7.4, 0.1 M) 0.5mL
Bacterial suspension 0.1mL

The Main Assay II was performed using a pre-incubation method. The components were added in turn to an empty test-tube:
Bacterial suspension 0.1mL
Test item solution 0.1mL
or control item solution 0.05mL
S9 mix or phosphate buffer (pH 7.4, 0.1 M) 0.5mL

The incubate was vortexed and placed at 37°C for 30 minutes. Two mL of overlay agar was then added and the mixture vortexed again and poured onto the surface of a minimal
medium agar plate and allowed to solidify.

The prepared plates were inverted and incubated for approximately 72 hours at 37°C. After this period of incubation, plates were immediately scored by counting the number of revertant colonies on each plate and evaluating the condition of the bacterial backgroundlawn. In the preliminary toxicity test, plates were held at approximately 4°C for 24 hours before scoring.
Evaluation criteria:
Four strains of Salmonella typhimurium (TA1535, TA1537, TA98 and TA100) and a strain of Escherichia coli (WP2 uvrA) were used in this study.
TA1535 and TA100 are predominantly sensitive to base pair mutagens, TA1537 and TA98 are sensitive to frameshift mutagens. In addition to amutation in the histidine operon, the Salmonella tester strains contain additional mutations which enhance their sensitivity to some mutagenic compounds. The rfa wall mutation results in the loss of one of the enzymes responsible for the synthesis of part of the lipopolysaccharide barrier that forms the surface of the bacterial cell wall and increases permeability to certain classes of chemicals. All strains are deficient in a DNA excision repair system (uvrB mutation) which enhances the sensitivity to some mutagens. TA98 and TA100 strains contain the pKM101 plasmid which activates an error prone DNA repair system.
Tester strain WP2 uvrA is reverted fromtryptophan dependence (auxotrophy) to tryptophan independence (prototrophy) by base substitution mutagens. In addition to the mutation in the tryptophan operon, the tester strain contains an uvrA DNA repair deficiency which enhances its sensitivity to some mutagenic.

The assay was considered valid if the following criteria were met:
1. Mean plate counts for untreated and positive control plates should fall within 2 standard deviations of the current historical mean values.
2. The estimated numbers of viable bacteria/plate should fall in the range of 100 – 500 millions for each strain.
3. No more than 5% of the plates should be lost through contamination or other unforeseen event.

For the test item to be considered mutagenic, two-fold (or more) increases in mean revertant numbers must be observed at two consecutive dose levels or at the highest practicable dose level only. In addition, there must be evidence of a dose-response relationship showing increasing numbers of mutant colonies with increasing dose levels.
Statistics:
as per OECD 471 guideline

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
Preliminary toxicity test:
No precipitation of the test item was observed at the end of the incubation period at any concentration tested, in the absence or presence of S9 metabolic activation. Neither toxicity, nor relevant increases in revertant numbers were observed with any tester strain at any dose level, in the absence or presence of S9 metabolism.

Main assay: No precipitation of the test item was observed at the end of the incubation period at any concentration in any experiment.
Neither toxicity, nor relevant increase in the number of revertant colonies was observed in the pre-incubation assay, at any dose level, with any tester strain, in the absence or presence of S9 metabolism.

Controls: Results show that mean plate counts for untreated and positive control plates fell within the normal range based on historical control data.

Controls of S9:
The sterility of the S9 mix and of the test item solutions was confirmed by the absence of colonies on additional agar plates spread separately with these solutions. Marked increases in revertant numbers were obtained in these tests following treatment with the positive control items, indicating that the assay system was functioning correctly.


Any other information on results incl. tables

Citoxicity and mutagenicity results are given in the following tables

BBL: Bacterial background lawn

A3689: Cytotoxicity - plate incorporation test without metabolic activation
Test item
(μg per plate)
TA98 TA100 TA1535 TA1537 E.Coli wp2 UVRa Bacteriotoxic effect Cytotoxicity
       Untreated  28 123 14 16 26 not evaluated no cytotoxicity
50.0 31 120 17 22 23 BBL: normal no cytotoxicity
158 31 110 16 19 27 BBL: normal no cytotoxicity
500 35 121 16 16 24 BBL: normal no cytotoxicity
1580 32 114 20 17 23 BBL: normal no cytotoxicity
5000 26 122 17 19 31 BBL: normal no cytotoxicity
               
Rt/Rc 50  1,11 0,98 1,21 1,38 0,88    
Rt/Rc 158 1,11 0,89 1,14 1,19 1,04    
Rt/Rc 500 1,25 0,98 1,14 1,00 0,92    
Rt/Rc 1580 1,14 0,93 1,43 1,06 0,88    
Rt/Rc 5000 0,93 0,99 1,21 1,19 1,19    
A3689: Cytotoxicity - plate incorporation test with metabolic activation
Test item
(μg per plate)
TA98 TA100 TA1535 TA1537 E.Coli wp2 UVRa Bacteriotoxic effect Cytotoxicity
       Untreated 37 136 16 24 36 not evaluated no cytotoxicity
50.0 35 114 14 18 39 BBL: normal no cytotoxicity
158 38 120 15 22 37 BBL: normal no cytotoxicity
500 33 115 14 22 35 BBL: normal no cytotoxicity
1580 36 127 18 17 37 BBL: normal no cytotoxicity
5000 33 123 16 21 40 BBL: normal no cytotoxicity
               
Rt/Rc 50  0,95 0,84 0,88 0,75 1,08    
Rt/Rc 158 1,03 0,88 0,94 0,92 1,03    
Rt/Rc 500 0,89 0,85 0,88 0,92 0,97    
Rt/Rc 1580 0,97 0,93 1,13 0,71 1,03    
Rt/Rc 5000 0,89 0,90 1,00 0,88 1,11    

A3689: Μutagenicity Experiment I - plate incorporation test - Strain: S. typhimurium TA 98
 without metabolic activation  with metabolic activation     Conclusion 
Test item
(μg per plate)
Revertants per plate mean s.e. Rt/Rc Bacteriotoxic effect Cytotoxicity Revertants per plate mean s.e. Rt/Rc Bacteriotoxic effect Cytotoxicity  
  Untreated  27 27 32 29 1,7 - not evaluated no cytotoxicity 32 33 32 32 0,3 - not evaluated no cytotoxicity valid
313 25 32 28 28 2,0 0,97 BBL: normal no cytotoxicity 37 38 39 38 0,6 1,19 BBL: normal no cytotoxicity not mutagenic
625 30 26 30 29 1,3 1,00 BBL: normal no cytotoxicity 32 39 41 37 2,7 1,16 BBL: normal no cytotoxicity
1250 36 29 26 30 3 1,03 BBL: normal no cytotoxicity 36 32 32 33 1,3 1,03 BBL: normal no cytotoxicity
2500 36 29 27 31 2,7 1,07 BBL: normal no cytotoxicity 31 33 36 33 1,5 1,03 BBL: normal no cytotoxicity
5000 28 25 26 26 0,9 0,90 BBL: normal no cytotoxicity 36 30 31 32 1,9 1,00 BBL: normal no cytotoxicity
2-Nitrofluorene/2-AA 186 161 179 175 7,4 6,48     435 408 444 429 10,8 12,62     valid
DMSO 26 28 28 27 0,7  -     35 33 33 34 0,7  -     valid
A3689: Μutagenicity Experiment I - plate incorporation test - Strain: S. typhimurium TA 100
 without metabolic activation  with metabolic activation Conclusion 
Test item
(μg per plate)
Revertants per plate mean s.e. Rt/Rc Bacteriotoxic effect Cytotoxicity Revertants per plate mean s.e. Rt/Rc Bacteriotoxic effect Cytotoxicity  
  Untreated  124 119 128 124 2,6 - not evaluated no cytotoxicity 123 123 125 124 0,7 - not evaluated no cytotoxicity valid
313 134 133 138 135 1,5 1,09 BBL: normal no cytotoxicity 161 161 148 157 4,3 1,27 BBL: normal no cytotoxicity not mutagenic
625 138 132 124 131 4,1 1,06 BBL: normal no cytotoxicity 146 153 163 154 4,9 1,24 BBL: normal no cytotoxicity
1250 139 121 134 131 5,4 1,06 BBL: normal no cytotoxicity 136 136 153 142 5,7 1,15 BBL: normal no cytotoxicity
2500 116 121 111 116 2,9 0,94 BBL: normal no cytotoxicity 118 142 113 124 9 1,00 BBL: normal no cytotoxicity
5000 131 139 132 134 2,5 1,08 BBL: normal no cytotoxicity 131 138 132 134 2,2 1,08 BBL: normal no cytotoxicity
AS/2AA 571 553 529 551 12,2 4,44     1286 1008 1412 1235 119,3 10,12     valid
DMSO            -     124 123 119 122 1,5  -     valid
A3689: Μutagenicity Experiment I - plate incorporation test - Strain: S. typhimurium TA 1535 
 without metabolic activation  with metabolic activation Conclusion 
Test item
(μg per plate)
Revertants per plate mean s.e. Rt/Rc Bacteriotoxic effect Cytotoxicity Revertants per plate mean s.e. Rt/Rc Bacteriotoxic effect Cytotoxicity  
  Untreated  13 16 14 14 0,9 - not evaluated no cytotoxicity 15 16 13 15 0,9 - not evaluated no cytotoxicity valid
313 15 18 16 16 0,9 1,14 BBL: normal no cytotoxicity 13 15 14 14 0,6 0,93 BBL: normal no cytotoxicity not mutagenic
625 12 18 14 15 1,8 1,07 BBL: normal no cytotoxicity 13 14 17 15 1,2 1,00 BBL: normal no cytotoxicity
1250 15 19 14 16 1,5 1,14 BBL: normal no cytotoxicity 14 18 18 17 1,3 1,13 BBL: normal no cytotoxicity
2500 18 15 19 17 1,2 1,21 BBL: normal no cytotoxicity 16 14 18 16 1,2 1,07 BBL: normal no cytotoxicity
5000 18 14 17 16 1,2 1,14 BBL: normal no cytotoxicity 20 14 15 16 1,9 1,07 BBL: normal no cytotoxicity
AS/2-AA 512 487 470 490 12,2 35,00     126 118 134 126 4,6 9,00     valid
DMSO            -     16 15 12 14 1,2  -     valid
A3689: Μutagenicity Experiment I - plate incorporation test - Strain: S. typhimurium TA 1537
 without metabolic activation  with metabolic activation Conclusion 
Test item
(μg per plate)
Revertants per plate mean s.e. Rt/Rc Bacteriotoxic effect Cytotoxicity Revertants per plate mean s.e. Rt/Rc Bacteriotoxic effect Cytotoxicity  
  Untreated  14 13 20 16 2,2 - not evaluated no cytotoxicity 22 21 24 22 0,9 - not evaluated no cytotoxicity valid
313 20 19 19 19 0,3 1,19 BBL: normal no cytotoxicity 18 18 16 17 0,7 0,77 BBL: normal no cytotoxicity not mutagenic
625 15 19 19 18 1,3 1,13 BBL: normal no cytotoxicity 16 19 20 18 1,2 0,82 BBL: normal no cytotoxicity
1250 15 17 20 17 1,5 1,06 BBL: normal no cytotoxicity 16 14 17 16 0,9 0,73 BBL: normal no cytotoxicity
2500 21 20 15 19 1,9 1,19 BBL: normal no cytotoxicity 22 20 19 20 0,9 0,91 BBL: normal no cytotoxicity
5000 14 15 18 16 1,2 1,00 BBL: normal no cytotoxicity 22 17 20 20 1,5 0,91 BBL: normal no cytotoxicity
9-AAc/2-AA 116 194 167 159 22,9 9,94     95 86 110 97 7 6,06     valid
DMSO 15 18 16 16 0,9  -     20 13 14 16 2,2  -     valid
A3689: Μutagenicity Experiment I - plate incorporation test - Strain: E. coli WP2 uvrA
 without metabolic activation  without metabolic activation Conclusion 
Test item
(μg per plate)
Revertants per plate mean s.e. Rt/Rc Bacteriotoxic effect Cytotoxicity Revertants per plate mean  s.e. Rt/Rc Bacteriotoxic effect Cytotoxicity  
  Untreated  26 30 27 28 1,2 - not evaluated no cytotoxicity 27 30 24 27 1,7 - not evaluated no cytotoxicity valid
313 25 31 33 30 2,4 1,07 BBL: normal no cytotoxicity 29 35 35 33 2 1,22 BBL: normal no cytotoxicity not mutagenic
625 26 30 27 28 1,2 1,00 BBL: normal no cytotoxicity 32 32 33 32 0,3 1,19 BBL: normal no cytotoxicity
1250 24 30 24 26 2 0,93 BBL: normal no cytotoxicity 31 32 35 33 1,2 1,22 BBL: normal no cytotoxicity
2500 28 24 24 25 1,3 0,89 BBL: normal no cytotoxicity 35 40 36 37 1,5 1,37 BBL: normal no cytotoxicity
5000 26 26 30 27 1,3 0,96 BBL: normal no cytotoxicity 39 32 38 36 2,2 1,33 BBL: normal no cytotoxicity
MMS/2-AA 168 187 164 173 7,1 6,18     195 227 204 209 9,5 7,21     valid
DMSO            -     27 31 30 29 1,2  -     valid

A3689: Μutagenicity Experiment II - pre-incubation test - Strain: S. typhimurium TA 98
 without metabolic activation  with metabolic activation Conclusion 
Test item
(μg per plate)
Revertants per plate mean  s.e. Rt/Rc Bacteriotoxic effect Cytotoxicity Revertants per plate mean s.e. Rt/Rc Bacteriotoxic effect Cytotoxicity  
Untreated 32 30 32 31 0,7 - not evaluated no cytotoxicity 37 38 35 37 0,9 - not evaluated no cytotoxicity valid
313 26 27 30 28 1,2 0,90 BBL: normal no cytotoxicity 37 35 38 37 0,9 1,00 BBL: normal no cytotoxicity not mutagenic
625 28 29 32 30 1,2 0,97 BBL: normal no cytotoxicity 36 39 27 34 3,6 0,92 BBL: normal no cytotoxicity
1250 29 32 34 32 1,5 1,03 BBL: normal no cytotoxicity 37 35 39 37 1,2 1,00 BBL: normal no cytotoxicity
2500 25 28 33 29 2,3 0,94 BBL: normal no cytotoxicity 39 39 36 38 1 1,03 BBL: normal no cytotoxicity
5000 30 31 28 30 0,9 0,97 BBL: normal no cytotoxicity 38 39 40 39 0,6 1,05 BBL: normal no cytotoxicity
2-Nitrofluorene/2-AA 187 175 199 187 6,9 6,93     513 462 481 485 14,9 12,76     valid
DMSO 28 28 26 27 0,7  -     35 39 40 38 1,5  -     valid
A3689: Μutagenicity Experiment II - pre-incubation test - Strain: S. typhimurium TA 100
 without metabolic activation  with metabolic activation Conclusion 
Test item
(μg per plate)
Revertants per plate mean  s.e. Rt/Rc Bacteriotoxic effect Cytotoxicity Revertants per plate mean ± sd s.e. Rt/Rc Bacteriotoxic effect Cytotoxicity  
Untreated 145 153 161 153 4,6 - not evaluated no cytotoxicity 162 169 153 161 4,6 - not evaluated no cytotoxicity valid
313 147 161 135 148 7,5 0,97 BBL: normal no cytotoxicity 184 165 167 172 2 1,07 BBL: normal no cytotoxicity not mutagenic
625 158 173 142 158 9 1,03 BBL: normal no cytotoxicity 171 154 158 161 5,1 1,00 BBL: normal no cytotoxicity
1250 143 148 151 147 2,3 0,96 BBL: normal no cytotoxicity 167 143 160 157 7,1 0,98 BBL: normal no cytotoxicity
2500 162 175 160 166 4,7 1,08 BBL: normal no cytotoxicity 172 166 153 164 5,6 1,02 BBL: normal no cytotoxicity
5000 159 143 146 149 4,9 0,97 BBL: normal no cytotoxicity 147 151 136 145 4,5 0,90 BBL: normal no cytotoxicity
AS/2AA 712 664 559 645 45,2 4,22     1232 1128 1346 1235 63 7,87     valid
DMSO            -     147 158 167 157 5,8  -     valid
A3689: Μutagenicity Experiment II - pre-incubation test - Strain: S. typhimurium TA 1535 
 without metabolic activation  with metabolic activation Conclusion 
Test item
(μg per plate)
Revertants per plate mean  s.e. Rt/Rc Bacteriotoxic effect Cytotoxicity Revertants per plate mean s.e. Rt/Rc Bacteriotoxic effect Cytotoxicity  
Untreated 13 13 15 14 0,7 - not evaluated no cytotoxicity 13 19 15 16 1,8 - not evaluated no cytotoxicity valid
313 12 14 13 13 0,6 0,93 BBL: normal no cytotoxicity 14 19 17 17 1,5 1,06 BBL: normal no cytotoxicity not mutagenic
625 18 19 16 18 0,9 1,29 BBL: normal no cytotoxicity 18 23 18 20 1,7 1,25 BBL: normal no cytotoxicity
1250 14 16 16 15 0,7 1,07 BBL: normal no cytotoxicity 21 18 19 19 0,9 1,19 BBL: normal no cytotoxicity
2500 13 16 14 14 0,9 1,00 BBL: normal no cytotoxicity 21 22 17 20 1,5 1,25 BBL: normal no cytotoxicity
5000 16 14 13 14 0,9 1,00 BBL: normal no cytotoxicity 16 17 15 16 0,6 1,00 BBL: normal no cytotoxicity
AS/2-AA 475 536 488 500 18,6 35,71     115 107 121 114 4,1 7,13     valid
DMSO                 17 14 18 16 1,2  -     valid
Μutagenicity Experiment II - pre-incubation test - Strain: S. typhimurium TA 1537
 without metabolic activation  with metabolic activation Conclusion 
Test item
(μg per plate)
Revertants per plate mean  s.e. Rt/Rc Bacteriotoxic effect Cytotoxicity Revertants per plate mean s.e. Rt/Rc Bacteriotoxic effect Cytotoxicity  
Untreated 17 15 20 17 1,5 - not evaluated no cytotoxicity 18 18 19 18 0,3 - not evaluated no cytotoxicity valid
313 15 18 22 18 2,0 1,06 BBL: normal no cytotoxicity 18 17 16 17 0,6 0,94 BBL: normal no cytotoxicity not mutagenic
625 15 16 17 16 0,6 0,94 BBL: normal no cytotoxicity 20 20 23 21 1 1,17 BBL: normal no cytotoxicity
1250 17 19 15 17 1,2 1,00 BBL: normal no cytotoxicity 19 19 20 19 0,3 1,06 BBL: normal no cytotoxicity
2500 16 20 15 17 1,5 1,00 BBL: normal no cytotoxicity 22 23 17 21 1,9 1,17 BBL: normal no cytotoxicity
5000 22 14 18 18 2,3 1,06 BBL: normal no cytotoxicity 24 24 25 24 0,3 1,33 BBL: normal no cytotoxicity
9-AAc/2-AA 184 116 153 151 19,7 10,07     89 104 93 95 4,5 4,75     valid
DMSO 13 14 17 15 1,2 -     17 19 23 20 1,8  -     valid
A3689: Μutagenicity Experiment II - pre-incubation test- Strain: E. coli WP2 uvrA
 without metabolic activation  without metabolic activation Conclusion 
Test item
(μg per plate)
Revertants per plate mean s.e. Rt/Rc Bacteriotoxic effect Cytotoxicity Revertants per plate mean  s.e. Rt/Rc Bacteriotoxic effect Cytotoxicity  
Untreated 26 28 23 26 1,5 - not evaluated no cytotoxicity 28 29 31 29 0,9 - not evaluated no cytotoxicity valid
313 24 23 22 23 0,6 0,88 BBL: normal no cytotoxicity 33 32 37 34 1,5 1,17 BBL: normal no cytotoxicity not mutagenic
625 27 24 26 26 0,9 1,00 BBL: normal no cytotoxicity 33 29 32 31 1,2 1,07 BBL: normal no cytotoxicity
1250 29 29 33 30 1,3 1,15 BBL: normal no cytotoxicity 32 32 32 32 0 1,10 BBL: normal no cytotoxicity
2500 27 29 26 27 0,9 1,04 BBL: normal no cytotoxicity 34 32 32 33 0,7 1,14 BBL: normal no cytotoxicity
5000 27 23 24 25 1,2 0,96 BBL: normal no cytotoxicity 33 29 35 32 1,8 1,10 BBL: normal no cytotoxicity
MMS/2-AA 142 138 135 138 2 5,31     227 223 215 222 3,5 6,73     valid
DMSO                 31 35 34 33 1,2  -     valid

Applicant's summary and conclusion

Conclusions:
Nut mutagenic in bacteria
Executive summary:

The mutagenicity potential in bacteria of Sodium (xylenes and 4-ethylbenzene) sulfonates was assessed following official guideline OECD 471, Bacterial Reverse Mutation Test. The test item does not induce reverse mutation in Salmonella typhimurium or Escherichia coli in the absence or presence of S9 metabolism, for all the tested strains under the reported experimental conditions.