Propane-1,3-diol

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

ACC PDO panel assigned reliability of 1. PDO from chemical process.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1994

Report date:

1994

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

First addendum to the OECD Guideline for Testing of Chemicals, Section 4, No; 471, adopted May 26, 1983

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EEC directive 92/69, L383 A, Annex V, B14, December 29, 1992

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Propane-1,3-diol

EC Number:

207-997-3

EC Name:

Propane-1,3-diol

Cas Number:

504-63-2

Molecular formula:

C3H8O2

IUPAC Name:

propane-1,3-diol

Details on test material:

- Name of test material (as cited in study report): 1,3-propanediol
- Physical state: liquid (colorless)
- Stability under test conditions: stable
- Storage condition of test material: room temperature
- Purity: ≥99.8%

Method

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

without S9 mix: 33.3; 100; 333.3; 1000; 2500; 5000 µg/plate
with S9 mix: 33.3; 100; 333.3; 1000; 2500; 5000 µg/plate

Vehicle / solvent:

distilled water

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

strain TA153, TA100; dissolved in distilled water; without metabolic activation

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Remarks:

without metabolic activation

Positive control substance:

other: 4-nitro-o-phenylene-diamine

Remarks:

strain TA 1537, TA 98; dissolved in DMSO

Controls 3

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

strain TA 102; dissolved in distilled water; without metabolic activation

Controls 4

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Remarks:

with metabolic activation

Positive control substance:

other: 2-aminoanthracene (2-AA)

Remarks:

All strains tested; dissolved in DMSO

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation - Experiment 1) and pre-incubation (Experiment 2)

Bacteria are stored as stock cultures in ampoules with nutrient broth + 5% DMSO in liquid nitrogen
Preculture: 10 hours in a shaking water bath at 37°C; thawed bacterial suspension in nutrient medium (8 g/L Merck nutrient broth and 5 g/L NaCl).

Selective agar: 2.0% Vogel-Bonner-Glucose-Minimal-Agar.
Overlay agar: 6.0 g/L Merck Agar Agar; 6.0 g/L NaCl; 10.5 mg/L L-histidine x HCl x H2O; 12.2 mg/L biotin.

Experimental performance:
Mixing the following components:
- 100 µL test solution at each dose level, positive or negative control
- 500 µL S9 mix or S9 mix substitution buffer
- 100 µL bacterial suspension
- 2000 µL overly agar
This mix was poured onto the selective agar plates; for the pre-incubation assay the mix was incubated at 37°C for 60 min prior to pouring onto minimal agar plates.

DURATION:
Exposure duration: After solidification at least 48h at 37°C in the dark (upside down).

NUMBER OF REPLICATIONS:
At least 3 replicates for each strain and dose level.

DATA RECORDING:
Colonies were counted using the AUTOCOUNT (Artek Systems Corporation, BIOSYS GmbH, Karben). If precipitation of the test substance precluded automatic counting the revertant colonies were counted by hand.

EVALUATION OF RESULTS:
- Corresponding background growth on both negative controls and test plates.
- Normal range of spontaneous reversion rates; normal frequencies are:
TA 1535: 10-29
TA 1537: 5-28
TA 98: 15-57
TA 100: 77-189
TA 102: 121-293

Evaluation criteria:

A test article in considered positive if either a dose related and reproducible increase in the number of revertants or a significant and reproducible increase for at least one test concentration is induced.
A test article producing neither a dose related and reproducible increase in the number of revertants nor a significant and reproducible positive response at any one of the test points is considered non-mutagenic in this system.

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA 1537, TA98, TA1535, TA 100, TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

no substantial increases in revertant colony numbers of any of the five tested strains (with or without metabolic activation)

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

tested up to 5000 µg/plate; with and without metabolic activation

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

The plates incubated with the test substance showed normal background growth up to 5000 µg/plate with and without S9 mix in all strains used.

Any other information on results incl. tables

Experiment I (Without Activation)	Mean Revertants Per Plate
	Strain  TA1535	Strain TA1537	Strain TA98	Strain TA100	Strain TA102
Negative Control	14	22	31	99	169
Solvent Control	19	19	26	99	186
1,3-Propanediol
33.3 µg/plate	13	24	25	100	184
100 3 µg/plate	17	23	30	93	164
333.3 µg/plate	21	15	26	102	179
1000 µg/plate	16	23	24	93	188
2500 µg/plate	13	22	26	97	194
5000 µg/plate	12	18	31	105	185
Positive control
Sodium azide(10 µg/plate)	885			619
4-NOPD (10 µg/plate)		62	245
MMS (5 µL/plate)					730

 

Experiment I (With Activation)	Mean Revertants Per Plate
	Strain  TA1535	Strain TA1537	Strain TA98	Strain TA100	Strain TA102
Negative Control	21	28	42	159	259
Solvent Control	18	23	40	158	244
1,3-Propanediol
33.3 µg/plate	16	21	45	163	240
100 3 µg/plate	18	26	45	160	236
333.3 µg/plate	22	29	50	156	255
1000 µg/plate	24	23	47	172	291
2500 µg/plate	20	18	43	156	274
5000 µg/plate	19	24	43	170	255
Positive control (2AA)
2.5 µg/plate	261	90	468	965	755

 

Experiment II (Without Activation)	Mean Revertants Per Plate
	Strain  TA1535	Strain TA1537	Strain TA98	Strain TA100	Strain TA102
Negative Control	7	22	24	79	153
Solvent Control	5	22	20	92	151
1,3-Propanediol
33.3 µg/plate	8	21	20	82	151
100 3 µg/plate	4	22	21	81	159
333.3 µg/plate	4	21	20	85	163
1000 µg/plate	7	22	19	69	151
2500 µg/plate	9	23	25	68	164
5000 µg/plate	9	19	25	83	148
Positive control
Sodium azide(10 µg/plate)	263			237
4-NOPD (10 µg/plate)		138	150
MMS (5 µL/plate)					874

 

Experiment II (With Activation)	Mean Revertants Per Plate
	Strain  TA1535	Strain TA1537	Strain TA98	Strain TA100	Strain TA102
Negative Control	10	30	20	137	185
Solvent Control	8	32	22	128	154
1,3-Propanediol
33.3 µg/plate	7	32	24	131	176
100 3 µg/plate	8	27	17	136	211
333.3 µg/plate	8	32	22	139	189
1000 µg/plate	14	31	25	125	182
2500 µg/plate	9	31	23	143	184
5000 µg/plate	14	30	20	126	195
Positive control (2AA)
2.5 µg/plate	39	143	197	406	611

Applicant's summary and conclusion

Conclusions:

Under the conditions of this study, propane-1,3-diol was non-mutagenic in a Salmonella typhimurium mutation assay in strains TA 1535, TA 1537, TA 98, TA 100 and TA 102 in the presence and absence of an exogenous metabolic activation system.

Executive summary:

The study was performed to investigate the potential of 1,3-propanediol to induce gene mutations according to the plate incorporation test (experiment I) and the pre-incubation test (experiment II) using the Salmonella typhimurium strains TA1535, TA1537, TA98, TA100, and TA102. The assay was performed in two independent experiments both with and without liver microsomal activation. Each concentration, including the controls, was tested in triplicate. Concentrations tested included: 0, 33.3, 100, 333.3, 1000, 2500, and 5000 µg/plate. No toxic effects occurred in the test groups with or without metabolic activation. The plates incubated with the test article showed normal background growth up to 5000 µg/plate with and without S9 mix in all strains used.

 

No substantial increases in revertant colony numbers of any of the five tester strains were observed following treatment with 1,3-propanediol at any dose level, either in the presence or absence of metabolic activation. There was also no tendency of higher mutation rates with increasing concentrations. The test substance did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used. Therefore, 1,3-propanediol was considered to be non-mutagenic in this Salmonella typhimurium reverse mutation assay.