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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
22 JUN 2016 to 28 JUL 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report date:
2016

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
dated 21 July 1997
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
GROUPE INTERMINISTERIEL DES PRODUITS CHIMIQUES
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
1,2-dimethyl-3-propyl imidazolium bis((trifluoromethyl)sulfonyl)amide
EC Number:
805-807-9
Cas Number:
169051-76-7
Molecular formula:
C10H15F6N3O4S2
IUPAC Name:
1,2-dimethyl-3-propyl imidazolium bis((trifluoromethyl)sulfonyl)amide
Test material form:
liquid
Details on test material:
Appearance : colorless oil
Composition: a base stock containing (C, 28.64; H, 3.61; F, 27.18; N, 10.02; O, 15.26; S, 15.29)
Molecular formula : C10H15F6N3O4S2
Molecular Weight : 419.12 g/mol
Purity >99% (purity determined by MNR)
Homogeneity : homogeneous
Specific details on test material used for the study:
Stability : stable under normal storage conditions
Batch production date: 07 JAN 2016
expiry date: 02 JAN 2030
Solubility : DMSO
PH : 6-7
Sterility : non sterile
Relative density : 1.47 Kg/L
storage conditions : room temperature -non hygroscopic

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 pKM 101
Metabolic activation:
with and without
Metabolic activation system:
S9-mix 10% (v/v). S9 fraction, microsome fraction prepared from Sprague Dawley rat liver homogenate, is provided by MOLTOX ( Boone, NC58607 - USA)
Test concentrations with justification for top dose:
5000, 1500, 500, 150, 50 µg/plate with and without Metabolic activation
The bacteriostatic activity test results show that the test item is soluble and non cytotoxic. So 5mg/Plate is the higher concentration chosen as the guidelines recommend.
Vehicle / solvent:
Solvent use : Dimethyl sulfoxide (DMSO).
Controls
Untreated negative controls:
yes
Remarks:
DMSO 50µl/Plate
Negative solvent / vehicle controls:
yes
Remarks:
Dimethyl sulfoxide (DMSO); Acetone; NaCl 0.15M 20µl/Plate
True negative controls:
yes
Remarks:
water 100µl/Plate
Positive controls:
yes
Positive control substance:
7,12-dimethylbenzanthracene
9-aminoacridine
2-nitrofluorene
sodium azide
other: 2-Anthramine; cis-platinum (II) Diammine dichloride
Details on test system and experimental conditions:
This study was performed in the absence and presence of metabolic activation. Two independent assays were carried.

Assay N°1 :
various concentrations of GBZ180716-S1 were put in contact with the strains in the absence and presence of metabolic activation system (S9-mix 10% (v/v)).
Method of application: in agar (plate incorporation).

Assay N°2 :
various concentrations of GBZ250716-S1 were put in contact with the strains in the absence of metabolic activation and with pre-incubation in the presence of metabolic activation system (S9-mix 10% (v/v)).
Method of application: without activation - plate incorporation; with activation - pre-incubation 30 min at 37° C.

Negative and positive controls were carried out in parallel.
After a 48-72h incubation period at 37°C, revertant colonies are counted in each plate.


Rationale for test conditions:
If the first assay is positive, the second one is performed in the same manner.
If the first assay, in the presence of test item is negative, the pre-incubation test is performed in the second assay.
Evaluation criteria:
- The bacteriostatic activity of the highest concentration tested shall be equal or less than 75 %,
- The spontaneous reversion rate of the absolute negative control shall comply with the historical values of the laboratory,
- The spontaneous reversion rate of the solvent shall not be statistically different from absolute negative control,
- The mean number of revertant colonies obtained for each strain and the corresponding positive control, with and/or without metabolic activation shall comply with the historical values of the laboratory
- Negative and positive values should not show significant difference with the historical values of the laboratory (± 2 standard deviation).
Statistics:
Data are presented as the number of revertant colonies (mean ± standard deviation) per plate. The following ratio is calculated:
R= Number of revertant colonies in the presence of the test item / Number of revertant colonies in the absence of the test item.

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
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
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
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
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
E. coli WP2 uvr A pKM 101
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
Results of sterility control of test item and S9-mix
For the two assays, negative and positive controls were carried out in parallel. Positive controls induced a significant increase in the number of revertant colonies compared to negative controls. There is no significant difference between the number of spontaneous reversions, the number of reversions obtained in the positive controls (without and with metabolic activation), and the mean of corresponding experimental "historical" values obtained in the laboratory. These results validate the two tests.
There is no evidence of any increase in the number of revertant colonies in the presence of the various concentration of the test item ( 5000, 1500, 500, 150 et 50 µg/plate), without and with metabolic activation in Salmonella typhimurium TA 1535, TA 1537, TA 98, TA 100 and in Escherichia coli WP2(uvrA-) (pKM 101).

Any other information on results incl. tables

Sterility control

Serie Doses Colony number/plate
Control n°1   1 2 3
Solution of
 
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium
bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

(LEMI code : GBZ180716-S1)		 5000 µg/plate 0 0 0
1500 µg/plate 0 0 0
500 µg/plate 0 0 0
150 µg/plate 0 0 0
50 µg/plate 0 0 0
S9-mix 500 µl/plate 0 0 0
Control n°2   1 2 3
Solution of
 
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

(LEMI code : GBZ250716-S1)		 5000 µg/plate 0 0 0
1500 µg/plate 0 0 0
500 µg/plate 0 0 0
150 µg/plate 0 0 0
50 µg/plate 0 0 0
S9-mix 500 µl/plate 0 0 0

Table 4 : sterility control

Bacterisostatic activiy controls

Doses (/plate)
  0
(negative control)		 DMSO 50 µg 150 µg 500 µg 1500µg 2500 µg 5000µg
Solution of
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

(LEMI code : GBZ180716-S1)		 N1 435 450 461 429 488 443 467 462
N2 484 479 483 453 452 467 473 475
N3 443 481 442 470 471 437 434 435
N 454± 26 470 ± 17 462 ± 21 451 ± 21 470 ± 17 470 ± 17 470 ± 17 470 ± 17
% - 1,04 1,02 0,99 1,04 0,99 1,01 1

Mutagenicity activity

TA1535 assay 1 -S9-mix

Serie Dose/plate Plate Mean Standard deviation R
1 2 3
Negative control 100 µl 12 11 13 12,00 1,00 _
positive solvent control 5 µl 10 7 9 8,67 1,53 _
Positive control :
Sodium azide		 5 µg
 in 5 µl		 595 658 576 609,67 42,92 70,35
Vehicle 50 µl 6 13 4 7,67 4,73 _
Solution of
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

LEMI code : GBZ180716-S1		 5000 µg 8 9 12 9,67 2,08 1,26
1500 µg 7 10 3 6,67 3,51 0,87
500 µg 13 11 7 10,33 3,06 1,35
150 µg 15 10 15 13,33 2,89 1,74
50 µg 18 13 8 13,00 5,00 1,70

TA1535 assay 1 10%S9-mix without pre-incubation

Serie Dose/plate Plate Mean Standard deviation R
1 2 3
Negative control 100 µl 16 10 8 11,33 4,16 _
positive solvent control 5 µl 16 10 9 11,67 3,79 _
Positive control :
2-Anthramine		 5 µg
 in 5 µl		 114 161 153 142,67 25,15 12,23
Vehicle 50 µl 15 15 8 12,67 4,04 _
Solution of
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

LEMI code : GBZ180716-S1		 5000 µg 13 10 9 10,67 2,08 0,84
1500 µg 10 17 22 16,33 6,03 1,29
500 µg 11 13 10 11,33 1,53 0,89
150 µg 11 10 12 11,00 1,00 0,87
50 µg 11 14 14 13,00 1,73 1,03

TA1535 Assay 2 -S9-mix

Serie Dose/plate Plate Mean Standard deviation R
1 2 3
Negative control 100 µl 8 14 8 10,00 3,46 _
positive solvent control 5 µl 9 14 6 9,67 4,04 _
Positive control :
Sodium azide		 5 µg
 in 5 µl		 653 890 889 810,67 136,54 83,86
Vehicle 50 µl 6 7 12 8,33 3,21 _
Solution of
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

LEMI code : GBZ180716-S1		 5000 µg 6 8 5 6,33 1,53 0,76
1500 µg 12 17 6 11,67 5,51 1,40
500 µg 8 8 11 9,00 1,73 1,08
150 µg 5 6 8 6,33 1,53 0,76
50 µg 4 6 9 6,33 2,52 0,76

TA1535 Assay 2 10%S9-mix with pre-incubation

Serie Dose/plate Plate Mean Standard deviation R
1 2 3
Negative control 100 µl 9 8 10 9,00 1,00 _
positive solvent control 5 µl 7 17 13 12,33 5,03 _
Positive control :
2-Anthramine		 5 µg
 in 5 µl		 50 60 50 53,33 5,77 4,32
Vehicle 50 µl 9 9 14 10,67 2,89 _
Solution of
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

LEMI code : GBZ180716-S1		 5000 µg 5 5 7 5,67 1,15 0,53
1500 µg 7 9 5 7,00 2,00 0,66
500 µg 12 12 13 12,33 0,58 1,16
150 µg 12 13 12 12,33 0,58 1,16
50 µg 8 9 10 9,00 1,00 0,84

TA1537 assay 1 -S9-mix

Serie Dose/plate Plate Mean Standard deviation R
1 2 3
Negative control 100 µl 6 7 5 6,00 1,00 _
positive solvent control 5 µl 9 11 12 10,67 1,53 _
Positive control :
9-Aminoacridine		 5 µg
 in 5 µl		 966 1274 1163 1134,33 155,99 106,34
Vehicle 50 µl 9 8 14 10,33 3,21 _
Solution of
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

LEMI code : GBZ180716-S1		 5000 µg 6 10 11 9,00 2,65 0,87
1500 µg 3 5 4 4,00 1,00 0,39
500 µg 10 7 7 8,00 1,73 0,77
150 µg 4 11 4 6,33 4,04 0,61
50 µg 9 14 8 10,33 3,21 1,00

TA1537 assay 1 10% S9 -mix without pre-incubation

Serie Dose/plate Plate Mean Standard deviation R
1 2 3
Negative control 100 µl 10 12 12 11,33 1,15 _
positive solvent control 5 µl 10 9 9 9,33 0,58 _
Positive control :
2-anthramine		 5 µg
 in 5 µl		 46 34 38 39,33 6,11 4,21
Vehicle 50 µl 7 10 13 10,00 3,00 _
Solution of
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

LEMI code : GBZ180716-S1		 5000 µg 8 9 7 8,00 1,00 0,80
1500 µg 7 15 14 12,00 4,36 1,20
500 µg 16 9 12 12,33 3,51 1,23
150 µg 12 17 9 12,67 4,04 1,27
50 µg 15 13 11 13,00 2,00 1,30

TA1537 Assay 2 -S9-mix

Serie Dose/plate Plate Mean Standard deviation R
1 2 3
Negative control 100 µl 14 9 10 11,00 2,65 _
positive solvent control 5 µl 8 9 14 10,33 3,21 _
Positive control :
9-Aminoacridine		 5 µg
 in 5 µl		 1553 1773 1500 1608,67 144,76 155,68
Vehicle 50 µl 11 15 12 12,67 2,08 _
Solution of
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

LEMI code : GBZ180716-S1		 5000 µg 5 4 6 5,00 1,00 0,39
1500 µg 7 6 5 6,00 1,00 0,47
500 µg 4 10 8 7,33 3,06 0,58
150 µg 5 7 9 7,00 2,00 0,55
50 µg 6 6 7 6,33 0,58 0,50

TA1537 Assay 2 10% S9-mix with pre-incubation

Serie Dose/plate Plate Mean Standard deviation R
1 2 3
Negative control 100 µl 12 4 6 7,33 4,16 _
positive solvent control 5 µl 18 8 7 11,00 6,08 _
Positive control :
2-anthramine		 5 µg
 in 5 µl		 35 43 50 42,67 7,51 3,88
Vehicle 50 µl 7 11 10 9,33 2,08 _
Solution of
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

LEMI code : GBZ180716-S1		 5000 µg 9 10 5 8,00 2,65 0,86
1500 µg 3 8 11 7,33 4,04 0,79
500 µg 15 7 10 10,67 4,04 1,14
150 µg 9 8 16 11,00 4,36 1,18
50 µg 12 15 10 12,33 2,52 1,32

TA98 assay 1 -S9-mix

Serie Dose/plate Plate Mean Standard deviation R
1 2 3
Negative control 100 µl 11 15 22 16,00 5,57 _
positive solvent control 5 µl 20 19 18 19,00 1,00 _
Positive control :
9-Aminoacridine		 5 µg
 in 5 µl		 285 223 258 255,33 31,09 13,44
Vehicle 50 µl 22 21 25 22,67 2,08 _
Solution of
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

LEMI code : GBZ180716-S1		 5000 µg 21 19 15 18,33 3,06 0,81
1500 µg 15 19 20 18,00 2,65 0,79
500 µg 20 15 19 18,00 2,65 0,79
150 µg 18 21 18 19,00 1,73 0,84
50 µg 23 16 13 17,33 5,13 0,76

TA98 assay 1 10%S9-mix without pre-incubation

Serie Dose/plate Plate Mean Standard deviation R
1 2 3
Negative control 100 µl 18 23 27 22,67 4,51 _
positive solvent control 5 µl 23 26 26 25,00 1,73 _
Positive control :
2-anthramine		 5 µg
 in 5 µl		 441 407 549 465,67 74,14 18,63
Vehicle 50 µl 23 35 27 28,33 6,11 _
Solution of
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

LEMI code : GBZ180716-S1		 5000 µg 15 26 16 19,00 6,08 0,67
1500 µg 22 26 19 22,33 3,51 0,79
500 µg 17 23 23 21,00 3,46 0,74
150 µg 25 12 24 20,33 7,23 0,72
50 µg 17 27 20 21,33 5,13 0,75

TA98 Assay 2 -S9-mix

Serie Dose/plate Plate Mean Standard deviation R
1 2 3
Negative control 100 µl 26 25 16 22,33 5,51 _
positive solvent control 5 µl 19 24 17 20,00 3,61 _
Positive control :
2-Nitrofluorene		 5 µg
 in 5 µl		 242 265 235 247,33 15,70 12,37
Vehicle 50 µl 25 19 29 24,33 5,03 _
Solution of
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

LEMI code : GBZ180716-S1		 5000 µg 21 18 12 17,00 4,58 0,70
1500 µg 21 21 24 22,00 1,73 0,90
500 µg 18 23 26 22,33 4,04 0,92
150 µg 16 24 19 19,67 4,04 0,81
50 µg 18 14 19 17,00 2,65 0,70

TA98 Assay 2 10%S9-mix with pre-incubation

Serie Dose/plate Plate Mean Standard deviation R
1 2 3
Negative control 100 µl 35 28 36 33,00 4,36 _
positive solvent control 5 µl 25 23 29 25,67 3,06 _
Positive control :
2-anthramine		 5 µg
 in 5 µl		 424 495 478 465,67 37,07 18,14
Vehicle 50 µl 35 28 34 32,33 3,79 _
Solution of
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

LEMI code : GBZ180716-S1		 5000 µg 31 27 19 25,67 6,11 0,79
1500 µg 33 25 22 26,67 5,69 0,82
500 µg 19 24 30 24,33 5,51 0,75
150 µg 20 30 29 26,33 5,51 0,81
50 µg 37 31 25 31,00 6,00 0,96

TA100 assay 1 -S9-mix

Serie Dose/plate Plate Mean Standard deviation R
1 2 3
Negative control 100 µl 52 75 64 63,67 11,50 _
positive solvent control 5 µl 87 53 42 60,67 23,46 _
Positive control :
Sodium azide		 5 µg
 in 5 µl		 973 1115 1103 1063,67 78,75 17,53
Vehicle 50 µl 71 68 80 73,00 6,24 _
Solution of
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

LEMI code : GBZ180716-S1		 5000 µg 58 63 78 66,33 10,41 0,91
1500 µg 63 51 88 67,33 18,88 0,92
500 µg 68 57 47 57,33 10,50 0,79
150 µg 65 61 58 61,33 3,51 0,84
50 µg 71 70 52 64,33 10,69 0,88

TA100 assay 1 -S9-mix without pre-incubation

Serie Dose/plate Plate Mean Standard deviation R
1 2 3
Negative control 100 µl 75 93 75 81,00 10,39 _
positive solvent control 5 µl 79 72 78 76,33 3,79 _
Positive control :
2-anthramine		 5 µg
 in 5 µl		 598 441 517 518,67 78,51 6,79
Vehicle 50 µl 85 70 78 77,67 7,51 _
Solution of
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

LEMI code : GBZ180716-S1		 5000 µg 83 88 69 80,00 9,85 1,03
1500 µg 111 96 92 99,67 10,02 1,28
500 µg 71 72 92 78,33 11,85 1,01
150 µg 68 73 74 71,67 3,21 0,92
50 µg 73 89 69 77,00 10,58 0,99

TA100 Assay 2 -S9-mix

Serie Dose/plate Plate Mean Standard deviation R
1 2 3
Negative control 100 µl 56 57 68 60,33 6,66 _
positive solvent control 5 µl 63 53 64 60,00 6,08 _
Positive control :
Sodium azide		 5 µg
 in 5 µl		 1010 984 1184 1059,33 108,74 17,66
Vehicle 50 µl 66 61 62 63,00 2,65 _
Solution of
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

LEMI code : GBZ180716-S1		 5000 µg 61 68 56 61,67 6,03 0,98
1500 µg 65 57 59 60,33 4,16 0,96
500 µg 64 42 50 52,00 11,14 0,83
150 µg 52 44 61 52,33 8,50 0,83
50 µg 53 46 48 49,00 3,61 0,78

TA100 Assay 2 10%S9-mix with pre-incubation

Serie Dose/plate Plate Mean Standard deviation R
1 2 3
Negative control 100 µl 63 72 92 75,67 14,84 _
positive solvent control 5 µl 77 80 77 78,00 1,73 _
Positive control :
2-anthramine		 5 µg
 in 5 µl		 509 333 491 444,33 96,84 5,70
Vehicle 50 µl 69 65 68 67,33 2,08 _
Solution of
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

LEMI code : GBZ180716-S1		 5000 µg 70 71 64 68,33 3,79 1,01
1500 µg 63 73 77 71,00 7,21 1,05
500 µg 77 82 74 77,67 4,04 1,15
150 µg 77 65 85 75,67 10,07 1,12
50 µg 66 78 76 73,33 6,43 1,09

E. COLI assay 1 -S9-mix

Serie Dose/plate Plate Mean Standard deviation R
1 2 3
Negative control 100 µl 145 149 118 137,33 16,86 _
positive solvent control 5 µl 170 152 143 155,00 13,75 _
Positive control :
cis-Platinum(II)		 5 µg
 in 5 µl		 394 248 353 331,67 75,30 2,14
Vehicle 50 µl 138 150 128 138,67 11,02 _
Solution of
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

LEMI code : GBZ180716-S1		 5000 µg 60 70 71 67,00 6,08 0,48
1500 µg 122 100 98 106,67 13,32 0,77
500 µg 115 129 140 128,00 12,53 0,92
150 µg 120 121 146 129,00 14,73 0,93
50 µg 153 156 128 145,67 15,37 1,05

E. COLI assay 1 -S9-mix without pre-incubation

Serie Dose/plate Plate Mean Standard deviation R
1 2 3
Negative control 100 µl 154 143 176 157,67 16,80 _
positive solvent control 5 µl 170 149 151 156,67 11,59 _
Positive control :
Dimethylbenzanthracene		 5 µg
 in 5 µl		 386 391 458 411,67 40,20 2,63
Vehicle 50 µl 169 155 190 171,33 17,62 _
Solution of
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

LEMI code : GBZ180716-S1		 5000 µg 93 115 131 113,00 19,08 0,66
1500 µg 135 141 151 142,33 8,08 0,83
500 µg 144 159 167 156,67 11,68 0,91
150 µg 160 161 163 161,33 1,53 0,94
50 µg 199 167 170 178,67 17,67 1,04

E. COLI Assay 2 -S9-mix

Serie Dose/plate Plate Mean Standard deviation R
1 2 3
Negative control 100 µl 156 154 143 151,00 7,00 _
positive solvent control 5 µl 151 146 164 153,67 9,29 _
Positive control :
cis-Platinum(II)		 5 µg
 in 5 µl		 301 307 344 317,33 23,29 2,07
Vehicle 50 µl 147 141 145 144,33 3,06 _
Solution of
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

LEMI code : GBZ180716-S1		 5000 µg 123 86 141 116,67 28,04 0,81
1500 µg 121 117 126 121,33 4,51 0,84
500 µg 128 102 119 116,33 13,20 0,81
150 µg 113 124 105 114,00 9,54 0,79
50 µg 109 125 143 125,67 17,01 0,87

E. COLI Assay 2 10%S9-mix with pre-incubation

Serie Dose/plate Plate Mean Standard deviation R
1 2 3
Negative control 100 µl 185 176 179 180,00 4,58 _
positive solvent control 5 µl 149 185 157 163,67 18,90 _
Positive control :
Dimethylbenzanthracene		 5 µg
 in 5 µl		 420 434 412 422,00 11,14 2,58
Vehicle 50 µl 189 192 155 178,67 20,55 _
Solution of
CAS 169051-76-7 / 1-Propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide
BATCH : Sy-16-0140

LEMI code : GBZ180716-S1		 5000 µg 86 144 98 109,33 30,62 0,61
1500 µg 99 162 148 136,33 33,08 0,76
500 µg 182 129 151 154,00 26,63 0,86
150 µg 160 149 181 163,33 16,26 0,91
50 µg 165 176 147 162,67 14,64 0,91

Historical control values

2006-2015 Without metabolic activation With metabolic activation (Without pre-incubation) With metabolic activation (with pre-incubation)
  nbr. of plates Mean min/max   nbr. of plates Mean min/max   nbr. of plates Mean min/max
                         
Salmonella typhimurium TA1535 Spontaneous rerversion 912 10.7± 3.4 4.0/23.0 Spontaneous rerversion 492 12.2± 4,0 3,0 / 23,0 Spontaneous rerversion 489 12.6± 4,1 5,0 /25,0
Sodium Azide 912 668.2± 202.4 45,0 / 1483,0 2-anthramine 492 95.2± 46,9 26,0 / 269,0 2-anthramine 489 69,9± 29,2 25,0 / 185,0
                         
Salmonella typhimurium TA1537 Spontaneous rerversion 912 5,8± 2,4 1,0 / 14,0 Spontaneous rerversion 492 7,9± 3,0 1,0 / 24,0 Spontaneous rerversion 489 8,2± 3,0 2,0 / 19,0
9-Aminoacridine 912 787,5± 385,0 219,0 / 1967,0 2-anthramine 492 53,6± 21,7 24,0 / 170,0 2-anthramine 489 49,8± 21,0 21,0 / 182,0
                         
Salmonella typhimurium TA98 Spontaneous rerversion 912 15,7± 3,7 6,0 / 29,0 Spontaneous rerversion 492 22,9± 4,6 12,0 /35,0 Spontaneous rerversion 489 23,0± 4,9 11,0 / 35,0
2-Nitrofluorene 912 532,2± 214,4 187,0 / 1667,0 2-anthramine 492 566,3± 195,1 219,0 / 1280,0 2-anthramine 489 479,4± 183,9 174,0 / 1370,0
                         
Salmonella typhimurium TA100 Spontaneous rerversion 912 65,2± 17,1 41,0 / 158,0 Spontaneous rerversion 489 108,3± 25,1 58,0 / 220,0 Spontaneous rerversion 489 108,4± 27,1 51,0 / 217,0
Sodium Azide 912 862,4± 270,3 381,0 / 1690,0 2-anthramine 489 898,1 ± 374,0 361,0 / 2163,0 2-anthramine 492 708,0± 295,9 309,0 / 1889,0
                         
Salmonella typhimurium TA102 Spontaneous rerversion 441 159,8± 37,6 58,0 / 285,0 Spontaneous rerversion 243 274,6± 50,0 149,0 / 384,0 Spontaneous rerversion 222 275,2± 48,4 159,0 / 402,0
Mitomycin C 441 748,0± 199,3 389,0 / 1739,0 Benzo[a]pyrene 243 1216,7± 409,1 490,0 / 3100,0 Benzo[a]pyrene 222 1191,1± 409,8 501,0 / 2700,0
                         
Escherichi coli WP2
(pKM 101) (uvrA-)		 Spontaneous rerversion 675 67,8± 20,0 40,0 / 165,0 Spontaneous rerversion 351 153,1± 28,9 80,0 / 264,0 Spontaneous rerversion 351 158,5± 31,4 69,0 / 250,0
cis-Platinium (II)
 Diamine Dichloride		 675 531,1± 160,5 280,0 / 1089,0 Dimethyl
benzanthracene		 351 739,1± 234,7 384,0 / 1680 Dimethyl
benzanthracene		 351 731,7± 271,3 405,0 /1680,0
                         
Escherichi coli WP2
(pKM 101)		 Spontaneous rerversion 12 48,8± 7,8 36,0 / 61,0 Spontaneous rerversion 6 56,2± 9,4 43,0 / 69,0 Spontaneous rerversion 6 58,7± 8,5 49,0 / 70,0
Mitomycin C 12 283,9± 48,3 189,0 / 367,0 Benzo[a]pyrene 6 192,3± 19,5 167,0 / 224,0 Benzo[a]pyrene 6 191,7± 19,7 167,0 / 220,0

Applicant's summary and conclusion

Conclusions:
Doses (5000, 1500, 500, 150 and 50 µg/plate) performed from solutions of the test item CAS 169051-76-7 / 1-propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide BATCH : Sy-16-0140, do not induce any mutagenic change in Salmonella typhimurium TA 1535, TA 1537, TA 98, TA 100 and Escherichia coli WP2(uvrA-) (pKM 101) without, or with metabolic activation, according to OECD Guideline n°471.
Executive summary:

Solutions ( GBZ180716-S1 and GBZ250716-S1) obtained from GBZ260516-1, have been tested for their capacity to induce reverse mutation in four Salmonella typhimurium strains and Escherichia coli WP2(uvrA-) (pKM 101) strain. This study was performed in absence or presence of metabolic activation. Two independant assays were caried out.

For assay n°1, various concentrations of GBZ180716-S1 were put in contact with the strains in the absence and presence of metabolic activation system (S9-mix 10% (v/v)).

Doses (5000, 1500, 500, 150 and 50 µg/plate) performed from solutions of the test item CAS 169051-76-7 / 1-propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide BATCH : Sy-16-0140, do not induce any mutagenic change in Salmonella typhimurium TA 1535, TA 1537, TA 98, TA 100 and Escherichia coli WP2(uvrA-) (pKM 101) without, or with metabolic activation, according to OECD Guideline n°471.

For assay n°2, various concentrations of GBZ250716-S1 were put in contact with the strains in the absence of metabolic activation and with pre-incubation in the presence of metabolic activation system (S9-mix 10% (v/v)).

For the two assays, negative and positive controls were carried out in parallel. Positive controls induced a significant increase in the number of revertant colonies compared to negative controls. There is no significant difference between the number of spontaneous reversions, the number of reversions obtained in the positive controls (without and with metabolic activation), and the mean of corresponding experimental "historical" values obtained in the laboratory. These results validate the two tests.

There is no evidence of any increase in the number of revertant colonies in the presence of the various concentration of the test item (5000, 1500, 500, 150 et 50 µg/plate) without and with metabolic activation in Salmonella typhimurium TA 1535, TA 1537, TA 98, TA 100 and in Escherichia coli WP2(uvrA-) (pKM 101).

Doses (5000, 1500, 500, 150 and 50 µg/plate) performed from solutions of the test item CAS 169051-76-7 / 1-propyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide BATCH : Sy-16-0140, do not induce any mutagenic change in Salmonella typhimurium TA 1535, TA 1537, TA 98, TA 100 and Escherichia coli WP2(uvrA-) (pKM 101) without, or with metabolic activation, according to OECD Guideline n°471.