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Toxicological information

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1980

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent

Method

Target gene:
No details
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium, other: TA100, TA 98, TA 1535, TA 1537 and TA 1538
Species / strain / cell type:
E. coli, other: WP2 uvr A and WP2 uvrA/pKM101
Metabolic activation:
with and without
Metabolic activation system:
SD rat (7 week old, male) liver, induced with PCB (KC-500)
Test concentrations with justification for top dose:
-S9 mix / +S9 mix ; 0, 100, 250, 500, 1,000, 2,500, 5,000 μg/plate
Vehicle / solvent:
Vehicle(s)/solvent(s) used: Water
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamid(-S9 mix: S. typhimurium TA100, TA98, E.Coli WP2 uvrA and WP2 uvrA/pKM101)
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
N-ethyl-N-nitro-N-nitrosoguanidine
Remarks:
(-S9 mix : S. typhimurium TA1535)
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
(-S9 mix : S. typhimurium TA1537)
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene (+S9 mix: S. typhimurium TA 100, TA 98 and TA 1535 and E. coli WP2 uvr A and WP2 uvrA/pKM101
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
benzo(a)pyrene
Remarks:
(+S9 mix : S. typhimurium TA 1537, 1538)
Details on test system and experimental conditions:
METHOD OF APPLICATION: preincubation


DURATION
Preincubation : 20 minutes
Temperature : 37 ˚C
Exposure duration: 2 days


NUMBER OF PLATES : Solvent / vehicle controls :3, Positive controls : 2, Test substance : 2

NUMBER OF TEST REPEAT : 2

Evaluation criteria:
A test substance is considered to be mutagenic when a dose-related increase in revertant colony count is observed and the number of revertant colonies per plate with the test substance is more than twice that of the negative control and when a reproducibility of test result is observed. Additionally a test substance for which the results do not meet the above criteria is considered non-mutagenic in this system.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with
Genotoxicity:
positive
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
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:
not examined
Positive controls validity:
valid
Species / strain:
S. typhimurium, other: S. typhimurium TA 100, TA 98, TA 1537, TA 1538
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:
not specified
Untreated negative controls validity:
not examined
Positive controls validity:
not specified
Species / strain:
E. coli, other: WP2 uvr A and WP2 uvrA/pKM101
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:
not specified
Untreated negative controls validity:
not examined
Positive controls validity:
not specified
Remarks on result:
other: strain/cell type: S. typhimurium TA 1535

Any other information on results incl. tables

Results of the bacterial reversion test of acrylonitrile [direct method: -S9]

Test substance dose (μg/plate) Revertant colonies per plate [Mean]
TA100 TA1535 TA98 TA1537 TA1538 WP2 uvrA WP2 uvrA / pKM101
Vehicle control 115, 135, 144

[131]
6, 7, 11

[8]
40, 48, 43

[44]
7, 10, 14

[10]
16, 19, 15

[17]
14, 11, 12

[12]
122, 144, 115

[127]
100 120, 148

[134]
7, 10

[9]
46, 46

[46]
13, 24

[19]
24, 21

[23]
15, 21

[18]
138, 137

[138]
250 120, 121

[121]
13, 11

[12]
27, 41

[34]
13, 7

[10]
17, 24

[21]
17, 18

[17]
104, 125

[115]
500 127, 142

[135]
9, 7

[8]
30, 40

[35]
17, 12

[15]
21, 19

[20]
13, 20

[17]
130, 123

[127]
1,000 146, 140

[143]
7, 10

[9]
39, 38

[39]
20, 21

[21]
33, 23

[28]
18, 12

[15]
117, 145

[131]
2,500 141, 155

[148]
8, 7

[8]
29, 48

[39]
13, 16

[15]
32, 23

[28]
23, 22

[23]
133, 149

[141]
5,000 124, 146

[135]
11, 10

[11]
37, 31

[34]
19, 21

[20]
16, 17

[17]
12, 22

[17]
140, 144

[142]
Positive control substance  AF-2 ENNG AF-2 9AAC 2NF AF-2 AF-2
μg/plate 0.01 5 0.1 80 2 0.01 0.005
colony/plate 565, 532

[549]
565, 743

[654]
479, 366

[423]
742, 607

[675]
281, 219

[250]
91, 104

[98]
938, 1010

[974]

AF-2: 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamid

ENNG : N-ethyl-N-nitro-N-nitrosoguanidine

9AAC : 9-aminoacridine

2NF : 2-nitrofluorene

Table2. Results of the bacterial reversion test of acrylonitrile [direct method: +S9]

Test substance dose (μg/plate) Revertant colonies per plate [Mean] 
TA100 TA1535 TA98 TA1537 TA1538 WP2 uvrA WP2 uvrA / pKM101
Vehicle control 127, 118, 128

[124]
16, 24, 15

[18]
45, 44, 36

[42]
11, 4, 5

[7]
39, 45, 33

[39]
9, 19, 19

[16]
214, 245, 240

[233]
100 117, 112

[115]
13, 23

[18]
55, 38

[47]
7, 6

[7]
36, 38

[37]
10, 15

[18]
220, 236

[228]
250 123, 125

[124]
22, 27

[25]
45, 43

[44]
5, 5

[5]
33, 34

[34]
24, 17

[21]
245, 218

[232]
500 114, 119

[117]
43, 28

[35]
44, 49

[47]
5, 7

[6]
41, 33

[37]
19, 15

[17]
235, 220

[228]
1,000 125, 138

[132]
46, 53

[50]
43, 44

[44]
7, 7

[7]
34, 44

[39]
16, 21

[19]
239, 264

[252]
2,500 121, 150

[136]
89, 62

[76]
26, 51

[39]
3, 6

[5]
35, 33

[34]
25, 24

[25]
279, 256

[268]
5,000 138, 120

[129]
73, 53

[63]
33, 47

[40]
4, 14

[9]
39, 31

[35]
15, 22

[19]
280, 291

[286]
Positive control substance  2AA 2AA 2AA B(a)P B(a)P 2AA 2AA
μg/plate 0.5 2 0.5 5 5 40 1
colony/plate 182, 213

[198]
97, 87

[92]
208, 209

[209]
98, 105

[102]
357, 428

[393]
409, 506

[458]
351, 354

[353]

2AA : 2-aminoanthracene

B(a)P : benzo(a)pyrene

Applicant's summary and conclusion

Conclusions:
Acrylonitrile was found to be mutagenic under the conditions of this study [positive: weak positive in TA1535 (+S9)] .
Executive summary:

The mutagenicity of acrylonitrile was examined using preincubation method in S. typhimurium TA 100, TA 98, TA 1535, TA 1537, TA 1538 and E. coli WP2uvr A and WP2 uvrA/pKM101, at concentrations up to 5000 μg/plate with or without a metabolic activation system. In this study, slight positive mutagenic responses were obtained in TA 1535 with metabolic activation. Therefore, acrylonitrile is considered to be mutagenic.