1,3-dichloropropan-2-ol

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:

comparable to guideline study

Data source

Reference

Reference Type:

publication

Title:

Salmonella mutagenicity tests IV Results from the testing of 300 chemicals

Author:

Zieger et al

Year:

1988

Bibliographic source:

Environ Mol. Mutagen 11( Suppl 12) 1-158

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

not specified

GLP compliance:

no

Remarks:

The study

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

1

Reference substance name:

1,3-dichloropropan-2-ol

EC Number:

202-491-9

EC Name:

1,3-dichloropropan-2-ol

Cas Number:

96-23-1

Molecular formula:

C3H6Cl2O

IUPAC Name:

1,3-dichloropropan-2-ol

Test material form:

liquid

Specific details on test material used for the study:

purchased from Aldrich chemical
Label purity 95%
analysed purity 94%

Method

Target gene:

HIstidine loci

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 97

Species / strain 3

Species / strain / cell type:

S. typhimurium TA 98

Species / strain 4

Species / strain / cell type:

S. typhimurium TA 100

Metabolic activation:

with and without

Metabolic activation system:

S9 fraction

Test concentrations with justification for top dose:

0 to 6666 µg/plate- The substance was initially tested in half log dose intervals upto the dose that elcited toxicity in the initial toxicity screen. At least 5 doses were tested in tripplicate.

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

9-aminoacridine

sodium azide

other: 4-nitro-o-phenylenediamine TA98 without metabolic activation; 2-aminoanthracene all strains with metabolic activaiton

Details on test system and experimental conditions:

Cultures were grown overnight with shaking at 37°C in Oxoid No. 2 broth, and their phenotypes were analyzed prior to their use for mutagenicity assays.
Preparation of Liver S-9 Fractions The S-9 (9,OOOg supernatant) fractions of Aroclor 1254-induced, male Sprague- Dawley rat and male Syrian hamster livers were prepared. The S-9 mixes were prepared immediately prior to use and contained 10% S-9; The substance was tested in the absence of metabolic activation and with rat and hamster S-9 fractions.

A preincubation assay was performed . The test chemical (0.05 ml), Salmonella culture (0.10 ml), and S-9 mix or buffer (0.50 ml) were incubated at 37"C, without shaking, for 20 min. Chemicals known or suspected to be volatile were incubated in capped tubes. The top agar was added and the contents of the tubes were mixed and poured onto the surface of petri dishes containing Vogel-Bonner medium. The histidine-independent (his') colonies arising on these plates were counted following two days incubation at 37°C. Plates were machine counted (New Brunswick, Edison, NJ; Artek, Farmingdale, NY) unless precipitate was present which interfered with the count, or the color of the test chemical on the plate reduced the contrast between the colonies and the background agar. At the discretion of the investigators, plates with low numbers of colonies were counted by hand.
1) Testing in strains TA97, TA98, TA100, and TA1535, 10% S-9 was used. 2) The first test of a chemical was without activation and with 10% S-9 in the S-9 mix. If a positive result was obtained the test was repeated. If the tests were negative they were repeated without S-9 and with 30% S-9. 3) The order of use of 10% and 30% S-9 was reversed. 4) Initial testing was in strains TA98 and TA100 without activation and with 30% rat and hamster S-9s. If a positive result was obtained in one of these two strains it was repeated and the other strains were not used. If the tests were negative, the other strains were used with 30% and 10% S-9. A chemical was not designated nonmutagenic unless it had been tested in strains TA98, TA100, TA1535, and TA97 and/or TA1537, without activation and with 10% and 30% rat and hamster $9. Occasionally, 5% S-9 was also used in all protocol variations.
All chemicals were tested initially in a toxicity assay to determine the appropri- ate dose range for the mutagenicity assay. The toxicity assay was performed using TA1OO. Toxic concentrations were those that produced a decrease in the number of his+ colonies, or a clearing in the density of the background lawn, or both.

Rationale for test conditions:

Standard for the reverese mutation assay

Evaluation criteria:

Evaluations were made at both the individual trial and overall chemical levels. Individual trials were judged mutagenic (+), weakly mutagenic (+ W), questionable (?), or nonmutagenic (-), depending on the magnitude of the increase of his+ revertants, and the shape of the dose-response. A trial was considered questionable (?) if the dose-response was judged insufficiently high to support a call of “ +W,” if only a single dose was elevated over the control, or if the increase seen was not dose related. During the study the standard evaluaiton critieria fro a positive result were not applied: 3 fold increase overcurrent solvent control for TA1535 and TA97 and 2 fold increase over solvent control for TA 98, TA100. Thes ehave been applied in the current analysis.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

positive

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 specified

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

positive

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 specified

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 97

Metabolic activation:

with

Genotoxicity:

positive

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 specified

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 97

Metabolic activation:

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 specified

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 98

Metabolic activation:

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 specified

Positive controls validity:

valid

Test results 6

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with

Genotoxicity:

positive

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 specified

Positive controls validity:

valid

Any other information on results incl. tables

Dose	TA100						TA1535						TA97						TA98
	no S9		Hamster S9		Rat S9		no S9		Hamster S9		Rat S9		no S9		Hamster S9		Rat S9		no S9		Hamster S9		Rat S9
µg/plate	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM
0	133	9.1	89	1.5	125	4	27	4.4	9	2.9	11	2.5	132	6.6	150	11.5	184	12.7	15	0.9	28	3.4	26	1.2
100	147	4.7	177	3.5	139	3.8	28	4.2	27	1.5	14	1.3	126	1.5	148	3.1	174	9.7	17	3.3	32	3.8	31	2.1
333	182	7.2	359	11.8	173	21.5	59	5.5	34	2.4	23	3.3	140	4.9	151	10.7	183	13.2	17	3.8	31	2.8	28	3.2
1000	272	7.5	725	13.5	247	20.9	195	12.8	218	18.9	59	7.9	138	10.5	259	11.7	209	4.4	19	4.2	39	0.6	40	6.2
3333	511	15.1	1936	30.7	545	9.4	517	12.6	525	12.7	236	15.8	151	11.8	485	10	227	8.7	23	4	59	3.1	37	1.5
6666	934	9.9	2350	18.5	852	34.4	828	12.9	734	29.6	324	9.2	148	7	749	12.2	265	3.5	20	1.5	64	3.3	49	7.2
POS	521	10.7	446	25.7	801	34.9	336	16.2	43	3.1	176	13.3	238	16.2	293	10.3	1438	46.7	157	9.6	143	9.4	247	8

Applicant's summary and conclusion

Conclusions:

Based upon the positive responses observed in all four strains tested, it can be concluded that the substance is a potential mutagen.