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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:
other: GLP, OECD N°471 Guideline (1983 May 26th)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1990
Report Date:
1990

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
No bacterial strain with an AT basepair at the primary reversion site is included
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
not specified
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): diisopropylamine
- Physical state: liquid
- Analytical purity: 99.81%
- Purity test date: 1989-09-07
- Lot/batch No.: 612.048.00.5
- Expiration date of the lot/batch: no data
- Storage condition of test material: stable in normal conditions of storage
- Other: Source of La Chambre

Method

Target gene:
Histidine operon
Species / strain
Species / strain / cell type:
other: TA1535, TA1537, TA1538, TA98, TA100
Metabolic activation:
with and without
Metabolic activation system:
Rat S9 mix. Liver S9 homogenate was prepared from rats that have been induced with Arochlor 1254.
Test concentrations with justification for top dose:
1st experiment: 100, 250, 500, 1000, 2500 ug/plate (TA1535, TA1537, TA1538, TA100); 50, 100, 250, 1000, 2500 ug/plate (TA98)
2nd experiment: 500, 1000, 2500, 5000, 10000 ug/plate (all strains)
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: water
- Justification for choice of solvent/vehicle: solubility
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: -S9: TA1535, TA100 sodium azide 1 µg/plate); TA1537 9-aminoacridine 100 µg/plate ; TA98, TA1535 2-nitrofluorene 2.5 µg/plate. +S9: 2-aminoanthracene
Details on test system and experimental conditions:
DETERMINATION OF CYTOTOXICITY (preliminary range-finding test)
- Test: in TA 98 and TA 100 strains, with or without S9 mix ; 6 dose-levels (one plate/dose level except control plate in triplicate): 50, 100, 500, 1000, 2500 and 5000µg/plate
- Method: relative total growth (decrease in the number of revertant colonies and/or a thinning of the bacterial lawn);

EXPERIMENTS
Number of independent experiments: 2.

METHOD OF APPLICATION:
* Direct plate incorporation method for preliminary test and first experiment
* Preincubation method for preliminary test and second experiment

DURATION
- Preincubation period: 20min
- Exposure duration: 48H

NUMBER OF REPLICATIONS: triplicates
Evaluation criteria:
Reproducible 2-fold increase in the number of revertant colonies compared with vehicle controls in any strain at any dose-level and/or evidence of a dose-relationship.
Statistics:
None

Results and discussion

Test results
Species / strain:
other: TA1535, TA1537, TA1538, TA98, TA100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
other: 1ste exp: 5000 µg/plate; 2e exp.: 10000 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

AMES TEST : Group results (without S-9 mix)
No. hist + revertant colonies/plate
    Strain TA1535 Strain TA1537 Strain TA1538 Strain TA98 Strain TA100
Group 0 : DISTILLED WATER n=3 mean 20.3 12.3 12.0 25.3 116.3
SEM 1.33 2.40 1.00 0.33 3.38
Group 1 : POSITIVE CONTROL n=2 mean 566.5 592.0 233.0 431.0 828.0
SEM 15.50 23.00 14.00 8.00 3.00
Group 2 : DIPA 50 µg/plate n=3 mean (0)* (0)* (0)* 20.7 (0)*
SEM     3.38  
Group 3 : DIPA 100 µg/plate n=3 mean 22.3 10.0 12.5 23.0 124.3
SEM 2.19 1.53 0.50 1.15 7.13
Group 4 : DIPA 250 µg/plate n=3 mean 22.7 10.7 13.3 29.7 142.7
SEM 3.38 4.18 2.03 3.18 4.10
Group 5 : DIPA 500 µg/plate n=3 mean 16.7 12.3 17.0 19.3 129.7
SEM 3.71 1.45 1.53 3.84 5.55
Group 6 : DIPA 1000 µg/plate n=3 mean 15.3 15.3 13.3 24.3 98.7
SEM 1.20 1.76 0.88 6.33 5.21
Group 7 : DIPA 2500 µg/plate n=3 mean 12.0 7.3 9.7 (0)* 46.7
SEM 2.00 1.45 2.40   15.21
* Concentration not tested            

AMES TEST : Group results (with S-9 mix)
No. hist + revertant colonies/plate
    Strain TA1535 Strain TA1537 Strain TA1538 Strain TA98 Strain TA100
Group 0 : DISTILLED WATER n=3 mean 16,3 11,3 22,3 28,0 128,0
SEM 2,03 1,45 3,53 3,797 6,35
Group 1 : POSITIVE CONTROL n=2 mean 210,5 112,5 164,5 871,5 597,5
SEM 0,50 6,50 3,50 11,50 38,50
Group 2 : DIPA 50 µg/plate n=3 mean (0)* (0)* (0)* 21,0 (0)*
SEM     1,73  
Group 3 : DIPA 100 µg/plate n=3 mean 18,0 14,3 23,7 28,7 148,0
SEM 2,52 0,67 2,33 0,67 6,08
Group 4 : DIPA 250 µg/plate n=3 mean 18,0 8,0 22,3 22,7 136,7
SEM 1,53 0,58 2,19 3,18 3,71
Group 5 : DIPA 500 µg/plate n=3 mean 16,3 12,0 23,3 27,3 133,3
SEM 2,19 3,06 2,60 8,09 19,03
Group 6 : DIPA 1000 µg/plate n=3 mean 18,3 11,3 23,0 27,7 143,3
SEM 1,76 0,88 4,36 2,40 1,76
Group 7 : DIPA 2500 µg/plate n=3 mean 14,7 13,3 15,3 (0)* 137,0
SEM 2,73 1,45 2,73   6,35
* Concentration not tested            

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

Diisopropylamine did not show any mutagenic activity in the bacterial reverse mutation test with Salmonella typhimurium.
Executive summary:

The potential of the test item Diisopropylamine (DIPA) to induce reverse mutation in Salmonella typhimurium was evaluated in accordance with the international guidelines (OECD 471, Commission Directive No. B13/14).

The test item was tested in two independent experiments, with and without a metabolic activation system, both performed according to the preincubation method and to the standard plate incorporation method.

Each strain TA1535, TA1537, TA1538, TA98 and TA100 was exposed to the test item at five dose-levels (three plates/dose-level) selected from a preliminary toxicity test: 100, 250, 500, 1000, 2500 µg/plate (TA1535, TA1537, TA1538, TA100) and 50, 100, 250, 1000, 2500 ug/plate (TA98) and then 500, 1000, 2500, 5000, 10000 µg/plate (all strains).

After 48 to 72 hours of incubation at 37°C, the revertant colonies were scored.

The test item did not induce any noteworthy increase in the number of revertants, both with and without S9 mix, in any of the five strains. Diisopropylamine did not show any mutagenic activity in the bacterial reverse mutation test with Salmonella typhimurium.