Diisopropylamine

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:

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 guideline

Test guideline 1

Qualifier:

according to guideline

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

Test guideline 2

Qualifier:

according to guideline

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

Test material information

Constituent 1

Reference substance name:

Diisopropylamine

EC Number:

203-558-5

EC Name:

Diisopropylamine

Cas Number:

108-18-9

Molecular formula:

C6H15N

IUPAC Name:

bis(propan-2-yl)amine

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:

S. typhimurium, 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.