Acrylonitrile

Genetic toxicity: in vitro

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

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Acrylonitrile

EC Number:

203-466-5

EC Name:

Acrylonitrile

Cas Number:

107-13-1

Molecular formula:

C3H3N

IUPAC Name:

prop-2-enenitrile

Method

Target gene:

No details

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium, other: TA100, TA 98, TA 1535, TA 1537 and TA 1538

Species / strain 2

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

Controls

Controls 1

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)

Controls 2

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)

Controls 3

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)

Controls 4

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

Controls 5

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 results

Test results 1

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

Test results 2

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

Test results 3

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

Test results 4

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.