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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:
From 1998-01-30 to 1998-3-31
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted in accordance with OECD guidelines and GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1998
Report date:
1998

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
Deviations:
no
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
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
Batch: 961113-2
Purity: 99.3wt%
Stability: Stable

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
preincubation with S9mix
Test concentrations with justification for top dose:
Range-finding test: 5, 20, 78, 313, 1250 and 5000 ug/plate (with and without S9mix)
Main test I and II:313, 625, 1250, 2500 and 5000 (with and without S9mix):
Vehicle / solvent:
DMSO
Justification: homogeneous suspension was obtained by using DMSO.
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
sodium azide
N-ethyl-N-nitro-N-nitrosoguanidine
other: 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide, 2-aminoanthrathene
Details on test system and experimental conditions:
METHOD OF APPLICATION:preincubation

DURATION
- Preincubation period: 20 minutes
- Exposure duration: 48 hours

NUMBER OF REPLICATIONS: 3


OTHER:
Evaluation criteria:
When more than two-fold increase of revertant colony compared to that in the negative control was observed, it is considered positive.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle 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
Vehicle controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1 Results of the Bacterial Reverse Mutation Study of the test substance (Dose-finding test)
 
With (+) or Testsubstance Number of revertants (number of colonies / plate)
without (-) concentration Base-pair change type Frameshift type
S9 mix (mg / plate) TA100 TA1535 WP2uvrA- TA98 TA1537
S9 mix       (-) Solvent control (DMSO) 120         13         12         16         6        
                   
5   115         11         16         14         8        
                   
20   120         9         16         6         4        
                   
78 121         10         20         18         8        
                   
313 115         4         19         17         8        
                   
1250 94         13         8         12         5        
                   
5000 117         14         16         11         7        
                   
S9 mix      (+) Solvent control (DMSO) 146         12         15         25         12        
                   
5   135         11         29         24         15        
                   
20   152         12         21         23         22        
                   
78 137         9         15         32         10        
                   
313 124         3         17         24         13        
                   
1250 179         10         15         28         14        
                   
5000 213         10         25         20         8        
                   
Positive control Name AF-2 NaN3 ENNG AF-2 9-AA
Concentration (mg / plate) 0.01 0.5 2 0.1 80
S9 mix (-) Number of revertants 671         185         465         251         664        
                   
Positive control Name 2-AA 2-AA 2-AA 2-AA 2-AA
Concentration (mg / plate) 1 2 10 0.5 2
S9 mix (+) Number of revertants 556         82         460         235         72        
                                                 
(Note) †: Precipitates was observed.             (Mean)
Solvent control : DMSO
            
AF-2: 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide, NaN3: sodium azide,
ENNG:N-ethyl-N'-nitro-N'-nitrosoguanidine, 9-AA: 9-aminoacridine hydrochloride, 2-AA: 2-aminoanthracene

Table 2 Results of the Bacterial Reverse Mutation Study of the test substance (Main Test 1)
 
With (+) or Testsubstance Number of revertants (number of colonies / plate)
without (-) concentration Base-pair change type Frameshift type
S9 mix (mg / plate) TA100 TA1535 WP2uvrA- TA98 TA1537
S9 mix       (-) Solvent control (DMSO) 116         10         13         18         4        
93 ( 107 ) 8 ( 7 ) 13 ( 12 ) 18 ( 15 ) 5 ( 4 )
112   12 ) 4   3 ) 11   1 ) 10   5 ) 3   1 )
313 129         6         14         6         1        
135 ( 124 ) 7 ( 6 ) 7 ( 10 ) 14 ( 10 ) 1 ( 1 )
108   14 ) 6   1 ) 10   4 ) 9   4 ) 2   1 )
625 114         9         12         12         2        
124 ( 122 ) 7 ( 8 ) 14 ( 11 ) 7 ( 12 ) 4 ( 2 )
127   7 ) 9   1 ) 6   4 ) 18   6 ) 1   2 )
1250 118         3         12         12         2        
119 ( 117 ) 7 ( 5 ) 5 ( 9 ) 9 ( 9 ) 2 ( 2 )
114   3 ) 4   2 ) 9   4 ) 6   3 ) 2   0 )
2500 110         9         14         9         2        
100 ( 104 ) 7 ( 6 ) 8 ( 9 ) 11 ( 11 ) 2 ( 2 )
102   5 ) 3   3 ) 6   4 ) 13   2 ) 2   0 )
5000 116         6         10         6         1        
116 ( 118 ) 5 ( 5 ) 11 ( 13 ) 16 ( 11 ) 1 ( 1 )
122   3 ) 3   2 ) 17   4 ) 11   5 ) 2   1 )
S9 mix      (+) Solvent control (DMSO) 156         14         21         23         5        
143 ( 151 ) 6 ( 9 ) 16 ( 17 ) 26 ( 24 ) 10 ( 6 )
154   7 ) 7   4 ) 15   3 ) 22   2 ) 2   4 )
313 136         9         11         18         7        
157 ( 147 ) 8 ( 9 ) 15 ( 13 ) 21 ( 17 ) 4 ( 5 )
148   11 ) 10   1 ) 14   2 ) 12   5 ) 3   2 )
625 152         9         19         20         6        
159 ( 160 ) 7 ( 8 ) 18 ( 16 ) 21 ( 22 ) 3 ( 8 )
168   8 ) 8   1 ) 10   5 ) 25   3 ) 15   6 )
1250 183         7         14         20         7        
148 ( 159 ) 11 ( 9 ) 23 ( 16 ) 19 ( 20 ) 5 ( 6 )
145   21 ) 8   2 ) 11   6 ) 21   1 ) 6   1 )
2500 163         10         15         20         3        
176 ( 169 ) 19 ( 14 ) 14 ( 13 ) 18 ( 20 ) 6 ( 4 )
168   7 ) 14   5 ) 9   3 ) 22   2 ) 2   2 )
5000 167         12         17         18         2        
187 ( 177 ) 4 ( 8 ) 16 ( 16 ) 23 ( 19 ) 2 ( 3 )
177   10 ) 9   4 ) 16   1 ) 16   4 ) 4   1 )
Positive control Name AF-2 NaN3 ENNG AF-2 9-AA
Concentration (mg / plate) 0.01 0.5 2 0.1 80
S9 mix (-) Number of revertants 642         188         535         226         642        
710 ( 674 ) 166 ( 180 ) 564 ( 573 ) 215 ( 221 ) 766 ( 693 )
669   34 ) 186   12 ) 619   43 ) 222   6 ) 671   65 )
Positive control Name 2-AA 2-AA 2-AA 2-AA 2-AA
Concentration (mg / plate) 1 2 10 0.5 2
S9 mix (+) Number of revertants 556         111         466         253         73        
544 ( 566 ) 109 ( 112 ) 477 ( 446 ) 282 ( 277 ) 75 ( 72 )
597   28 ) 117   4 ) 394   45 ) 296   22 ) 69   3 )
(Notes) †: Precipitates was observed.             (Mean)
(±S.D.)
AF-2: 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide, NaN3: sodium azide,
ENNG:N-ethyl-N'-nitro-N'-nitrosoguanidine, 9-AA: 9-aminoacridine hydrochloride, 2-AA: 2-aminoanthracene

Table 3 Results of the Bacterial Reverse Mutation Study of the test substance (Main Test 2)
 
With (+) or Testsubstance Number of revertants (number of colonies / plate)
without (-) concentration Base-pair change type Frameshift type
S9 mix (mg / plate) TA100 TA1535 WP2uvrA- TA98 TA1537
S9 mix       (-) Solvent control (DMSO) 127         10         18         17         4        
123 ( 128 ) 18 ( 14 ) 25 ( 20 ) 13 ( 12 ) 5 ( 4 )
133   5 ) 14   4 ) 18   4 ) 6   6 ) 3   1 )
313 122         10         21         9         3        
122 ( 115 ) 12 ( 11 ) 17 ( 17 ) 5 ( 9 ) 7 ( 4 )
100   13 ) 12   1 ) 13   4 ) 12   4 ) 1   3 )
625 127         11         20         9         4        
126 ( 121 ) 12 ( 12 ) 26 ( 23 ) 13 ( 10 ) 6 ( 4 )
111   9 ) 12   1 ) 23   3 ) 7   3 ) 1   3 )
1250 117         13         24         8         4        
125 ( 117 ) 11 ( 13 ) 16 ( 19 ) 11 ( 12 ) 2 ( 3 )
108   9 ) 14   2 ) 16   5 ) 16   4 ) 3   1 )
2500 88         10         17         13         4        
129 ( 115 ) 10 ( 10 ) 15 ( 17 ) 7 ( 10 ) 4 ( 3 )
128   23 ) 9   1 ) 20   3 ) 11   3 ) 2   1 )
5000 117         18         19         7         2        
90 ( 114 ) 12 ( 13 ) 17 ( 20 ) 5 ( 6 ) 5 ( 3 )
136   23 ) 10   4 ) 24   4 ) 7   1 ) 3   2 )
S9 mix      (+) Solvent control (DMSO) 138         19         23         19         8        
121 ( 133 ) 13 ( 15 ) 24 ( 24 ) 26 ( 22 ) 6 ( 6 )
139   10 ) 13   3 ) 24   1 ) 21   4 ) 4   2 )
313 144         22         25         17         6        
160 ( 149 ) 17 ( 18 ) 27 ( 25 ) 25 ( 20 ) 3 ( 5 )
142   10 ) 16   3 ) 22   3 ) 18   4 ) 6   2 )
625 138         24         29         19         8        
138 ( 154 ) 22 ( 20 ) 24 ( 27 ) 16 ( 17 ) 6 ( 7 )
185   27 ) 13   6 ) 29   3 ) 17   2 ) 7   1 )
1250 192         25         24         17         6        
180 ( 191 ) 18 ( 22 ) 29 ( 26 ) 22 ( 19 ) 3 ( 5 )
201   11 ) 23   4 ) 25   3 ) 19   3 ) 7   2 )
2500 175         22         23         25         3        
186 ( 175 ) 14 ( 18 ) 31 ( 29 ) 17 ( 20 ) 4 ( 5 )
165   11 ) 19   4 ) 32   5 ) 19   4 ) 8   3 )
5000 178         17         33         23         8        
175 ( 178 ) 22 ( 19 ) 19 ( 24 ) 19 ( 19 ) 4 ( 6 )
182   4 ) 17   3 ) 19   8 ) 15   4 ) 7   2 )
Positive control Name AF-2 NaN3 ENNG AF-2 9-AA
Concentration (mg / plate) 0.01 0.5 2 0.1 80
S9 mix (-) Number of revertants 651         181         403         238         571        
689 ( 682 ) 156 ( 168 ) 451 ( 412 ) 248 ( 235 ) 615 ( 646 )
705   28 ) 168   13 ) 382   35 ) 220   14 ) 752   94 )
Positive control Name 2-AA 2-AA 2-AA 2-AA 2-AA
Concentration (mg / plate) 1 2 10 0.5 2
S9 mix (+) Number of revertants 495         104         382         248         86        
513 ( 512 ) 100 ( 101 ) 332 ( 340 ) 235 ( 241 ) 80 ( 83 )
529   17 ) 99   3 ) 306   39 ) 241   7 ) 84   3 )
(Notes) †: Precipitates was observed.             (Mean)
(±S.D.)
AF-2: 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide, NaN3: sodium azide,
ENNG:N-ethyl-N'-nitro-N'-nitrosoguanidine, 9-AA: 9-aminoacridine hydrochloride, 2-AA: 2-aminoanthracene

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

The test substance was negative for mutagenic activity.