1-(N,N-bis(2-hydroxyethyl)amino)propan-2-ol

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

Study period:

18 Jan to 12 Feb 1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD guideline study to GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

1-(N,N-bis(2-hydroxyethyl)amino)propan-2-ol

EC Number:

229-764-5

EC Name:

1-(N,N-bis(2-hydroxyethyl)amino)propan-2-ol

Cas Number:

6712-98-7

Molecular formula:

C7H17NO3

IUPAC Name:

1-[bis(2-hydroxyethyl)amino]propan-2-ol

Test material form:

other: Transparent, colourless viscous liquid

Details on test material:

- Name of test material (as cited in study report): Diethanolisopropanolamine (DEIPA)
- Substance type: Technical product
- Physical state: Liquid
- Analytical purity: 93%
- Impurities (identity and concentrations): No data
- Composition of test material, percentage of components: No data
- Isomers composition: No data
- Purity test date: Not specified
- Lot/batch No.: 980035B
- Expiration date of the lot/batch: Not specified
- Stability under test conditions: Not specified
- Storage condition of test material: Not specified

Method

Target gene:

S. typhimurium: Histidine locus
E. coli WP2 uvr A: Tryptophan locus

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

S9 fraction from Aroclor-induced rat liver

Test concentrations with justification for top dose:

100, 330, 1000, 3330, 5000 μg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: water
- Justification for choice of solvent/vehicle: Not specified [OECD guideline recommends the use of aqueous solvent/vehicle]

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

2-nitrofluorene

sodium azide

benzo(a)pyrene

other: 2-aminoanthracene and ICR-191

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation

Preincubated in the presence of the tester strain for 20 mins at 37oC before adding the overlay agar and pouring onto the surface of minimal agar plates. The plates were incubated for 52 +/- 4 hours at 37 +/- 2oC.

DURATION
- Preincubation period:20 +/- 2 minutes
- Exposure duration: 48-56 hours

SELECTION AGENT (mutation assays): Deficiency in histidine or tryphtophan for selection of non-amino acid requiring mutants.

NUMBER OF REPLICATIONS: 2 independant assays

DETERMINATION OF CYTOTOXICITY
- Method: A decrease in the number of revertant colonies per plate and/or by a thinning or disappearance of the bacterial background lawn indicates cytotoxicity

Evaluation criteria:

The test article had to produce at least a 3-fold (TA98, TA1535, TA1537 and WP2uvr A) or 2-fold (TA100) dose related and reproducible increase in the mean revertants per plate of at least one tester strain over the mean revertants per plate of the appropriate vehicle control

Statistics:

None reported

Results and discussion

Test results

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

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
Not applicable

RANGE-FINDING/SCREENING STUDIES: Ten doses of test article from 6.67 to 5000 ul per plate were tested. No cytotoxicity observed with TA100 and WP2uvrA either in presence or absence of metabolic activation system, normal backgroud lawn evident and no decrease in number of revertants per plate. No test article precipitate observed at any of the doses tested.

COMPARISON WITH HISTORICAL CONTROL DATA: Revertants were within the range of historical data for the vehicle and positive controls with each of the strains tested, both with and without S9.

ADDITIONAL INFORMATION ON CYTOTOXICITY: None reported

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

In a GLP study, conducted according to OECD guideline 471, DEIPA showed no convincing evidence of mutagenic activity when tested at concentrations of up to 5000 µg/plate in a bacterial reverse mutation assay with Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and Escherichia coli strain WP2uvrA, with and without metabolic activation (S9).

Executive summary:

In a GLP study, conducted according to OECD guideline 471, DEIPA was assessed for mutagenicity in a bacterial reverse mutation assay.

Two independent studies using the preincubation method were performed using Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and Escherichia coli strain WP2uvrA in the presence and absence of a rat liver metabolic activation (S9 mix). Concentrations of 100, 333, 1000, 3330, and 5000 µg/plate of DEIPA were tested, along with concurrent positive and vehicle controls, all plated in triplicate.The positive control substances all showed mutagenic activity, demonstrating that the test was valid.

Following incubation of the test material, tester strain and S9 mix (or phosphate buffer), the number of revertant colonies per plate was counted. Cytotoxicity was evaluated based on the condition of the bacterial background lawn. No precipitation of the test article was obseved at any dose level.

Under the conditions of this study, DEIPA did not cause a positive increase in the number of revertant colonies per plate and no cytotoxicity was evident, with any of the tester strains, at any concentration, either in the presence or absence S9.

DEIPA showed no convincing evidence of mutagenic activity.