Propane-1,2-diyl diacetate

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:

30 January - 21 February 1986

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to test guidelines and in accordance with GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1986

Report date:

1986

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Propane-1,2-diyl diacetate

EC Number:

210-817-6

EC Name:

Propane-1,2-diyl diacetate

Cas Number:

623-84-7

Molecular formula:

C7H12O4

IUPAC Name:

propane-1,2-diyl diacetate

Details on test material:

Purity >99.5%

Method

Target gene:

Strain Histidine mutation Mutation type
TA1537 his C3076 Frameshift
TA1538 his D3052 Frameshift
TA98 his D3052/R-factor* Frameshift
TA1535 his G46 Base-pair substitutions
TA100 his G46/R- factor * Base-pair substitutions
*R-factor = plasmid pKM101 (increases error-prone DNA repair).

Each tester strain contains the following additional mutations:
rfa: deep rough (defective lipopolysaccharide cell coat).
gal: mutation in the galactose metabolism.
chl: mutation in nitrate reductase.
bio: defective biotin synthesis.
uvrB: loss of the excision repair system (deletion of the ultraviolet-repair B gene).

Species / strain

Species / strain / cell type:

other: Strains TA98, TA100, TA1535, TA1537 and TA1538

Metabolic activation:

with and without

Metabolic activation system:

S9 from Aroclor 1254 induced male Wistar rat liver

Test concentrations with justification for top dose:

100, 333, 1000, 3333 and 5000 ug/plate without S9
100, 333, 1000, 3333 and 5000 ug/plate with S9

Vehicle / solvent:

reagent water

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

reagent water

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: Without S9: TA1535-sodium azide, TA100-methylmethanesulfonate, TA98 and TA1538-4-nitro-o-phenylenediamine, TA1537-9aminoacridine; With S9: all strains-2-aminoanthracene

Details on test system and experimental conditions:

The Salmonella tester strains have been provided by Dr. Bruce N. Ames, University of California at Berkeley, USA.

Samples of frozen stock cultures of bacteria are transferred into enriched nutrient broth (Oxoid No.2) and incubated in a shaking water bath (37C, 150 spm) until the cultures reach an 0.D. of 0.4 at 700 nm ( 10(9) cells/ml). Freshly grown cultures of each strain are used for a test.

Standard plate test
Top agar in top agar tubes is melted and heated to 45°C. The following solutions are successively added to 3 ml of top agar: 0.1 ml of a fresh bacterial culture (10(9) cells/ml) of one of the tester strains, 0.1 ml of a dilution of the test substance in reagent grade water, and in the case of activation assays 0.5 ml of S9-mix. The ingredients are mixed on a Vortex and the contents of the top agar tube are poured onto a selective agar plate. After solidification of the top agar, the plates are.turned and incubated in the dark at 37°C for 48h. After this period revertant colonies (histidin independent) are counted automatically with an Artek model 880 colony counter or manually.

Evaluation criteria:

No additional information available.

Statistics:

No additional information available.

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA98, TA100, TA1535, TA1537 and TA1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

In a preliminary test with strain TA100, eleven serial dilutions of the test substance, in approximately half-log steps, were plated with a diluted TA100 culture on non-selective agar (viability counting). For viability determinations, equal numbers of bacterial cells were seeded on each plate in the presence of the test substance. The percentage survival of an appropriately diluted TA100 culture on non-selective agar is determined by comparing the number of colonies on the solvent control plate with those on the plates containing the test substance. However, even at the highest test substance
concentration used the survival of strain TA100 is not reduced. Based on these data, the test substance was tested up to a concentration of 5 mg/plate, which is about the maximum test substance concentration that should be used, according to the OECD guidelines.

The Ames Salmonella microsome plate test
All bacteri al strains showed negative responses over the entire dose range of the test substance, i. e. no statistically significant dose-related increase in the number of revertants. Strain-specific positive control chemicals showed that the test conditions were optimal and that the metabolic activation system functioned properly. Based on these results, the test substance can be considered as nonmutagenic in the Ames Salmonella/microsome assay.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1 MUTAGENIC RESPONSE OF 1,2 DIACETOXY-PROPANE IN THE AMES SALMONELLA/MICROSOME

PLATE TEST

Experiment 1

	Mean number of revertant (His+) colonies/3 replicate plates (+S.D) with different strains of S. typhimurium
Dose (ug/plate)	TA1535	TA1537	TA1538	TA98	TA100
	Without S9 -mix
100	11 + 3	11 + 3	18 + 4	39 + 2	105 + 14
333	12 + 1	9 + 3	18 + 4	48 + 3	115 + 11
1000	11 + 6	13 + 4	21 + 4	46 + 4	129 + 13
3333	12 + 1	11 + 1	17 + 4	48 + 3	118 + 17
5000	13 + 6	12 + 1	18 + 2	43 + 6	111 + 3
Solvent control^a)	10 + 5	13 + 1	19 + 2	39 + 3	118 + 8
Positive control	275 + 33	1410 + 125	1985 + 108	2365 + 192	1207 + 74
	With S9 -mix
100	13 + 6	10 + 3	28 + 2	43 + 5	111 + 7
333	11 + 2	12 + 5	30 + 3	38 + 5	122 + 2
1000	10 + 4	11 + 3	25 + 2	39 + 10	126 + 2
3333	12 + 2	12 + 3	26 + 6	40 + 5	124 + 6
5000	10 + 1	12 + 4	26 + 6	34 + 5	125 + 5
Solvent controla)	11 + 3	11 + 6	27 + 4	43 + 4	121 + 2
Positive control	169 + 3	123 + 26	833 + 45	1330 + 27	1887 + 86

^a) 0.1 ml reagent grade water.

Table 2 MUTAGENIC RESPONSE OF 1,2 DIACETOXY-PROPANE IN THE AMES SALMONELLA/MICROSOME

PLATE TEST

Experiment 2

	Mean number of revertant (His+) colonies/3 replicate plates (+S.D) with different strains of S. typhimurium
Dose (ug/plate	TA1535	TA1537	TA1538	TA98	TA100
100	12 + 5	18 + 2	19 + 3	32 + 5	168 + 64
333	10 + 3	15 + 6	13 + 3	30 + 6	105 + 12
1000	12 + 6	19 + 5	22 + 4	32 + 8	110 + 24
3333	9 + 3	22 + 3	13 + 3	35 + 5	116 + 13
5000	9 + 1	14 + 4	17 + 5	28 + 3	138 + 30
Solvent control^a)	8 + 4	15 + 5	16 + 5	31 + 5	109 + 9
Positive control	264 + 55	1678 + 85	1237 + 35	1198 + 87	773 + 41
	With S9-mix
100	10 + 1	17 + 3	19 + 1	28 + 5	111 + 26
333	7 + 2	15 + 4	21 + 6	28 + 5	108 + 16
1000	9 + 1	16 + 5	18 + 2	36 + 8	123 + 4
3333	11 + 2	13 + 2	24 + 2	33 + 7	124 + 14
5000	12 + 2	15 + 3	22 + 7	36 + 9	108 + 6
Solvent control^a)	11 + 1	16 + 1	16 + 2	32 + 5	b)
Positive control	141 + 8	69 + 10	873 + 94	1111 + 168	1999 + 52

^a) 0.1 ml reagent grade water.

^b) Inadvertently not measured

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

The test substance induced no statistically significant dose related increase in the numbers of revertant (His+) colonies in any of the five tester strains (TA 1535; TA 1537; TA 1538; TA 98; TA 100) with or without metabolic activation. The test substance can therefore be considered as non mutagenic in this test system.

Executive summary:

PGDA (propylene glycol diacetate) was tested in the Ames Salmonella/microsome test up to the limit of toxicity (5 mg/plate).

The test substance induced no statistically significant dose related increase in the numbers of revertant (His+) colonies in any of the five tester strains (TA 1535; TA 1537; TA 1538; TA 98; TA 100). The test substance can therefore be considered as non mutagenic in this test system.