Registration Dossier

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
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Tests were performed at Microbiological Associates Inc. and SRI International. The study is not done according to OECD guideline. But it is a well documented study.

Data source

Referenceopen allclose all

Reference Type:
publication
Title:
Salmonella mutagenicity tests: IV. Results from the testing of 300 chemicals.
Author:
Zeiger E., Anderson B., Haworth S., Lawlor T. and Mortelmans K.
Year:
1988
Bibliographic source:
Environ Mol Mutagen. Suppl 12:1-157
Reference Type:
secondary source
Title:
ACToR: Aggregated Computational Toxicology Resource
Author:
EPA U.S. environmental protection agency
Year:
2013
Bibliographic source:
http://actor.epa.gov/actor/faces/ACToRHome.jsp
Report Date:
2013

Materials and methods

Principles of method if other than guideline:
Ames testSalmonella typhimurium strains TA97, TA98, TA100, TA1535, and TA1537 were obtained from Dr. Bruce Ames (University of California, Berkeley) and were stored as recommended (Maron and Ames, 1983). Cultures were grown overnight with shaking at 37°C in Oxoid No. 2 broth, and their phenotypes were analyzed prior to their use for mutagenicity assays. The S-9 (9,OOOg supernatant) fractions of Aroclor 1254-induced, male Sprague-Dawley rat and male Syrian hamster livers were prepared as described previously (Haworth et al, 1983). The S-9 mixes were prepared immediately prior to use and contained either 10% or 30% S-9; occasionally, other levels were used. All chemicals were tested in the absence of metabolic activation and with rat and hamster S-9 fractions.The preincubation assay was performed as described previously (Haworth et al., 1983), with some differences, as described below. The test chemical (0.05 ml), Salmonella culture (0.1 ml) and S-9 mix or buffer (0.5 ml) were incubated at 37°C, without shaking, for 20 min. The top agar was added and the contents of the tubes were mixed and poured onto the surface of petri dishes containing Vogel-bonner medium. The histidine-independent colonies arising on these plates were counted following two days incubation at 37°C. Plates were machine counted (New Brunswick, NJ; Artek, Farmingdale, NY) unless precipitate was present which interfered with the count, or the color of the test chemical on the plate reduced the contrast between the colonies and the background agar. At the discretion of the investigators, plates with low numbers of colonies were counted by hand. At least five doses of each chemical were tested in triplicate. Experiments were repeated at least one week following the initial trial. A maximum of 0.05 ml solvent was added to each plate. Concurrent solvent and positive controls were run with each trial.
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
not specified
Details on test material:
- Name of test material (as cited in study report): Quinidine- other: obtained from Fluka Chemical

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 100
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
S. typhimurium TA 1535
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
S. typhimurium TA 97
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
S. typhimurium TA 98
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
male Syrian Hamster, Liver, S-9, Aroclor 1254 (10%)
Test concentrations with justification for top dose:
33 - 3333 µg/PlateIn the first lab (Microbiological Associates Inc, MIC) 0 µg/Plate, 33 µg/Plate, 100 µg/Plate, 333 µg/Plate, 1000 µg/Plate and 2000 µg/Plate were used. In the second lab (SRI International) 0 µg/Plate, 33 µg/Plate, 100 µg/Plate, 333 µg/Plate, 1000 µg/Plate, 1666 µg/Plate and 3333 µg/Plate were used.
Vehicle / solvent:
Vehicle(s)/solvent(s) used: DMSO
Controls
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
9-aminoacridine
sodium azide
other: 4-nitro-o-phenylenediamine, 2-aminoanthracene
Remarks:
Positive controls in the absence of metabolic activation were sodium azide (TA1535 and TA100), 9-aminoacridine (TA97 and TA1537), and 4-nitro-o-phenylenediamine (TA98). Positive control for metabolic activation with all strains was 2-aminoanthracene.
Details on test system and experimental conditions:
METHOD OF APPLICATION: preincubation

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
2000 µg/plate quinidine
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
2000 µg/plate quinidine
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 97
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
2000 µg/plate quinidine
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
2000 µg/plate quinidine
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Remarks on result:
other: strain/cell type: Ames Salmonella typhimurium
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

1. Lab: MIC (Microbiological Associates), Solvent: DMSO

          TA100    TA1535  TA97     
 Dose (µg/Plate)  NA  10 % HLI  10% RLI

NA

  10 % HLI   10% RLI   NA   10 % HLI   10% RLI
 0 108 +0.9 88 +5.5 109 +5.6 36 +0.3 12 +1.9 12 +4.0 100 +5.7 145 +8.2 142 + 6.8
33 105 +3.1 106 +4.3 115 +6.0 42 +3.5 7 +1.2 15 +2.2 96 +13.7 131 +1.8 142 +8.6
 100 91 +4.4 101 +5.2 105 +4.1 44 +1.5 15 +2.1 13 +1.3 97 +5 .9 154 +4 .9 130 +4.1
 333 102 +9.1 96 +2.0 112 +5.3 56 +0.6 14 +1.8 12 +1.3 93 +8.0 161 +7.5 111 +3.9
 1000 103 +10.7 103 +7.0 115 +5.7 36s +2.8 13 +4.2  12s +1.8 71s +8.4 171 +4.7  131s +6.6 
 2000  t 120s +2.9 125 +9.3 32s +4.0 11s +0.3 11s +1.8  t 185s +22.8 116s +17.2
 POS 922 +26.3 1042 +105.6 1625 +17.6 1199 +18.7  71 +3.1  97 +4.2 1147 +118.5 760 +12.7 1058 +33.1

    TA98    
 Dose (µg/Plate)  NA  5 % HLI  10 % HLI

30 % HLI

  10 % RLI
 0 20 +2.0 32 +1.5 26 +1.5 28 +4.9 45 +2.5
33 18 +1.9 32 +2.4 34 +3.2 28 +4.9 32 +2.0
 100 18 +4.6 36 +4.7 30 +2.6 28 +4.6 37 +1.5
 333 21 +0.9 33 +0.3 32 +4.3 31 +2.2 42 +5.0
 1000 20s +1.2 31 +4.3 26 +7.5 33 +1.9 41 +2.3
 2000  18s +4.5 29s +2.6 41s +3.2 38s +3.0 51s +1.5
 POS 2047 +34.3 2376 +13.6 1571 +87.7 1552 +37.5  1819 +8.7 

2. Lab: SRI (SRI International), Solvent: DMSO

 

    TA100         

  Dose (µg/Plate)   NA   10 % HLI  30 % HLI   10 % RLI   30 % RLI
0  112 +5.5  115 +0.9  135 +7.1  118 +9.7  133 +4.0
33  102 +3.5  135 +12.5  115 +10.5  107 +4.3  127 +6.7
100  120 +3.5  124 +14.8  114 +4.9  115 +8.2  136 +2.5
333  133 +14.7  148 +0.6  138 +7.3  131 +8.4  131 +6.0
1000  114 +7.4  112 +10.8  139 +15.1  115 +7.2  140 +11.9
1666      131 +4.2    134 +11.3
3333  0s +0.3  1s +0.9    0s +0.0  
POS  757 +53.9  2419 +26.3  585 +12.0  1034 +34.2  380 +27.6

    TA1535         

  Dose (µg/Plate)   NA   10 % HLI  30 % HLI   10 % RLI   30 % RLI
0  18 +0.6  8 +2.3  10 +2.2  6 +0.6  13 +2.3
33  25 +3.3  7 +2.6  11 +0.9  5 +1.2  14 +1.9
100  21 +2.6  7 +0.7  3 +0.5  8 +1.0  19 +5.0
333  21 +4.4  6 +0.9  8 +2.9  7 +1.9  16 +5.8
1000  15 +0.3  4 +1.2  9 +1.5  2 +0.6  14 +1.5
1666      10 +3.3    10 +2.0
3333  1s +0.6  0s +0.0    0s +0.3  
POS  596 +19.1  539 +31.1  408 +4.6  310 +9.3  118 +2.0

    TA97         

  Dose (µg/Plate)   NA   10 % HLI  30 % HLI   10 % RLI   30 % RLI
0  133 +11.0  162 +5.0  168 +9.6  186 +12.4  169 +7.8
33  149 +5.5  193 +9.9  151 +28.9  195 +3.8  180 +37.3
100  134 +20.0  203 +10.2  153 +18.6  191 +9.6  215 +32.5
333 164 +5.5  208 +7.4  203 +10.4  201 +1.8  170 +5.8
1000  145 +8.4  165 +24.5  199 +8.3  152 +11.1  212 +6.2
1666      196 +18.3    132 +26.4
3333  3s +1.2  1s +0.6    17s +4.3  
POS  1844 +77.2  2255 +36.6  1097 +20.9  1479 +49.5  475 +19.9

    TA98 

  Dose (µg/Plate)   NA   10 % HLI  30 % HLI   10 % RLI   30 % RLI
0  19 +3.2  24 +2.3  28 +3.9  16 +1.7  28 +2.1
33 12 +0.3  26 +1.3  20 +4.6  18 +3.2  28 +6.3
100  16 +2.1  17 +2.0  23 +1.8  21 +2.0  28 +2.3
333  14 +0.6  22 +3.6  22 +3.0  14 +0.9  28 +3.7
1000  18 +2.1  12 +3.8  22 +3.5  8 +0.7  28 +3.4
1666      28 +5.5    29 +5.2
3333  1s +0.9  0s +0.0    1s +0.7  
POS  1489 +52.9  1444 +57.2  391 +17.7  513 +25.8  207 +18.7

NA: not activated; HLI: Aroclor 1254-induced hamster liver S-9; RLI: Aroclor 1254-induced rat liver S-9; t: complete clearing of background lawn (colonies not counted); s: slight clearing of background lawn

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):negative with metabolic activationnegative without metabolic activationThe genetic toxicity of quinidine was determined using the Ames test with and without metabolic activation. For the four Salmonella typhimurium strains TA 100, TA 1535, TA 97 and TA 98 no mutations were detected. From the result it can be concluded, that quinidine has no mutagenic effects.
Executive summary:

In the in vitro study publication by Zeiger et al., 1988 the genotoxicity of quinidine was determined with the Ames Test on the four Salmonella typhimurium strains TA 100, TA 1535, TA 97 and TA 98 with and without metabolic activation. For all strains no mutations were detected up to 3333 µg quinidine per plate. Therefore, we can be conclude, that quinidine is not genotoxic.