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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study without detailed documentation

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1995
Report date:
1995

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Qualifier:
according to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Bis(2-propylheptyl) phthalate
EC Number:
258-469-4
EC Name:
Bis(2-propylheptyl) phthalate
Cas Number:
53306-54-0
Molecular formula:
C28H46O4
IUPAC Name:
1,2-bis(2-propylheptyl) benzene-1,2-dicarboxylate
Test material form:
liquid

Method

Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254-induced rat liver S9-mix
Test concentrations with justification for top dose:
20, 100, 500, 2500 and 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Acetone
Controlsopen allclose all
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene; 2.5 µg dissolved in DMSO for TA 100, TA 98, TA 1537 and TA 1535
Remarks:
with metabolic activation
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
N-ethyl-N-nitro-N-nitrosoguanidine
Remarks:
without metabolic activation
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 4-nitro-o-phenylendiamine; 10 µg dissolved in DMSO for TA 98
Remarks:
without metabolic activation
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
without metabolic activation Migrated to IUCLID6: 100 µg dissolved in DMSO for TA 1537
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation) and preincubation


DURATION
- Preincubation period: 20 min, 37°C
- Exposure duration: 48 h, 37°C


NUMBER OF REPLICATIONS: 3


DETERMINATION OF CYTOTOXICITY
- Method: relative total growth
Evaluation criteria:
In general, a substance to be characterized as positive in the Ames test has to fulfill the following requirements:
- doubling of the spontaneous mutation rate (control)
- dose-response relationship
- reproducibility of the results

Results and discussion

Test results
Key result
Species / strain:
S. typhimurium, other: TA1535, TA1537, TA98, TA100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: A precipitation was found from about 500 µg/plate onward
Remarks on result:
other: all strains/cell types tested

Any other information on results incl. tables

Results standard plate assay:

Dose µg/plate metabolic activation  TA98   TA100   TA1535   TA1537 
Trial 1 Trial 2 Trial 3 Trial 1 Trial 2 Trial 3 Trial 1 Trial 2 Trial 3 Trial 1 Trial 2 Trial 3
0 + 50 38 38 128 118 123 15 20 24 14 17 10
20 + 46 35 30 106 117 122 14 15 26 15 15 18
100 + 34 27 49 130 127 101 21 17 14 8 19 8
500 + 40 31 30 108 111 117 9 22 20 11 9 16
2500 + 34 35 51 147 125 152 10 - 11 10 11 7
5000 + 32 33 40 144 124 125 14 11 18 11 16 8
           
2.5 µg 2-Aminoanthracene + 628 591 599 1080 997 1076 146 80 92 81 93 102
           
0 - 33 31 31 118 144 129 17 22 13 13 12 14
20 - 18 35 33 132 153 127 10 20 29 13 13 13
100 - 31 27 33 128 127 - 17 11 24 16 17 11
500 - 34 25 36 120 120 137 16 17 17 9 11 14
2500 - 40 36 28 142 138 167 24 28 21 9 13 8
5000 - 33 29 27 157 155 162 19 20 20 9 7 8
             
5 µg MNNG -   1560 1700 1997 1514 1650 1345
10 µg 4-Nitro-o-phenylendiamin - 640 533 386    
100 µg 9-amninoacridine -                   371 430 541

Preincubation test:

Dose µg/plate metabolic activation  TA98   TA100   TA1535   TA1537 
Trial 1 Trial 2 Trial 3 Trial 1 Trial 2 Trial 3 Trial 1 Trial 2 Trial 3 Trial 1 Trial 2 Trial 3
0 + 48 46 31 112 98 95 16 15 19 9 6 11
20 + 49 48 43 88 118 106 14 16 14 14 15 9
100 + 32 47 45 73 91 95 18 27 17 9 9 4
500 + 42 48 47 110 86 81 11 20 14 11 11 10
2500 + 47 59 49 104 91 92 4 12 18 9 6 13
5000 + 53 48 56 94 93 108 20 13 18 7 10 9
           
2.5 µg 2-Aminoanthracene + 599 649 624 585 529 687 92 102 108 82 76 80
           
0 - 49 55 47 136 95 96 19 19 20 13 11 11
20 - 50 60 35 105 134 103 15 23 22 8 12 5
100 - 35 56 43 103 100 104 26 16 22 13 16 12
500 - 38 47 41 107 112 110 25 19 16 7 13 9
2500 - 43 32 47 89 94 100 15 23 17 6 12 14
5000 - 50 49 38 122 121 100 12 23 20 12 12 15
           
5 µg MNNG - 555 690 629 911 825 780    
10 µg 4-Nitro-o-phenylendiamin - 1340 1460 1450        
100 µg 9-amninoacridine -                   581 692 519

Applicant's summary and conclusion