1,3-dichloropropan-2-ol

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

1,3 -Dichloropropanol has been found to be mutagenic in numerous strains of Salmonella typhimurium in the Ames test (Ziegler et al; Hahn et al), further in vitro testing was not conducted.

Link to relevant study records

Reference

Reference 1

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

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

Specific details on test material used for the study:

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

Target gene:

HIstidine loci

Species / strain / cell type:

S. typhimurium TA 1535

Species / strain / cell type:

S. typhimurium TA 97

Species / strain / cell type:

S. typhimurium TA 98

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

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.

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

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

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

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

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

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

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

Conclusions:

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

Reference 2

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

only 2 strains (TA100 and TA 1535 used), No plate incorporation assay conducted.

GLP compliance:

no

Remarks:

Data from openliterature study

Target gene:

histidne loci

Species / strain / cell type:

S. typhimurium TA 1535

Species / strain / cell type:

S. typhimurium TA 100

Metabolic activation:

with and without

Metabolic activation system:

rat S9

Test concentrations with justification for top dose:

1-62.8 µmol/plate (TA100)
1-78.6 µmol/plate (TA 1535)

Vehicle / solvent:

DMSO

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

sodium azide

other: 2-aminoanthracene

Details on test system and experimental conditions:

The 30-min preincubation test was performed according to Maron and Ames
using S~ typhimurium strain TA100 and strain TA1535, kindly provided by Professor B.N. Ames, with and without addition of
S9 mix (30.5 mg protein ml-1) prepared from livers of Wistar rats pretreated
with arochlor. The mean values of at least
two independent duplicate determinations were calculated. Differences in colony
counts were in all cases less than 20%. Colonies were counted using a Biotran
III automatic colony counter. The substances were dissolved in dimethyl sulphoxide
(10 µl). The average number of spontaneous revertants per plate was 190 for
strain TA100 and 22 for TA1535. 660 revertants per µg NaN3 per plate were
determined in the positive control for strain TA100 and 600 for TA1535. 1161
revertants per plate were obtained as positive control in the presence of S9 mix
with 2-aminoanthracene (1 µg per plate).

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

The results of the mutagenicity testing of 1,3-DCP-OH with S. typhimurium TA100 in the 30-min preincubation assay are shown in Table I. There are no significant differences between the results obtamed with and without addition of the metabolizing enzymes S9 mix or two different concentrations of ADH and cofactor NAD +. In order to examine whether the ADH present in S9 mix could cause activation, we added the cofactor NAD ÷ instead of NADP ÷ to S9 in an additional set of experiments (Table I). Here too no significant differences were observed. When using the S. typhimurium test strain TA1535 (Table II) a clear decrease in revertants was observed after addition of $9 mix or $9 mix and ADH whereas

addition of ADH and cofactor NAD ÷ without S9 mix did not influence the revertant numbers. In principle, an activating effect by ADH is possible and could be demonstrated with 2-chloropropenol. We have recently shown that 2-chloropropenol is metabolically transformed to 2-chloropropenal, an extremely strong mutagen . A comparison of the mutagenicity of 1,3-dichloroacetone and 1,3-DCP-OH shows that 1,3-dichloroacetone possesses a much higher activity than 1,3-DCP-OH. We found 23 875 revertants/#mol for 1,3-dichloroacetone in strain TA100 and 1137 in strain TA1535 but only about 33 revertants for 1,3-DCP-OH in both strains. Therefore activation should be

also expected if ADH converts 1,3-DCP-OH to 1,3-dichloroacetone. An activation by ADH and cofactors was, however, not observed in this case. In further mutagenicity studies we investigated the influence of the incubation time and whether preincubation of 1,3-DCP-OH without bacteria, in analogy to the SOS chromotest (see later), also leads to activation. A 90-min preincubation without S9 mix resulted only in marginally higher mutagenicity than was obtained with a 30-min preincubation. An additional incubation of the substance without bacteria for 30 min before the 90 rain preincubation with bacteria had also practically no effect.

TABLE I MUTAGENICITY OF 1,3-DCP-OH IN S. TYPHIMURIUM TA100 WITH AND WITHOUT S9 MIX AND WITH ADH IN REVERTANTS/PLATE AFTER 30-MIN PREINCUBATION

µmol/plate	-S9	+S9 NADP+	+S9 NAD+	ADH + NAD+ 5µl	ADH +NAD+ 50µl
1	239	258	245	224	210
2.6	241	281	281	248	282
5.2	323	375	326	324	358
7.9	351	441	379	371	360
10.5	418	480	418	431	395
15.7	484	559	473	499	504
26.2	630	696	589	689	609
36.7	793	843
52.4	818	889	812	792	796
62.8	827	743	822	781	835

TABLE II MUTAGENICITY OF 1,3-DCP-OH IN S. TYPHIMURIUM TA1535 WITH OR WITHOUT ADDITION OF THE METABOLIZING ENZYMES $9 MIX OR ADH IN REVERTANTS/PLATE AFTER 30 MIN PREINCUBATION

µmol/plate	-S9	+S9 NADP+	ADH + NAD+ 5µl	ADH +NAD+ 50µl
1	69	33	40
2.6	119	56	63	114
5.6	222	88	85	227
7.9	305	137	144	295
10.5	317	137	168	328
26.2	506	277	278	503
52.4	631	435	402	607
62.8		366	431

Conclusions:

Exposure of TA100 and TA1535 strains of salmonella to the substance results in a clear positive result in the reverse mutaiton assay.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (positive)

Genetic toxicity in vivo

Description of key information

Subsequent to the positive results found in the Ames test, standard somatic testing in mouse bone marrow erythrocytes did not demonstrate genotoxic potential.Similarly no genotoxic potential was found in a Drosophila spp wing spot test. A COMET assay conducted by oral gavage in mice found a positive dose dependant indication of genotoxicity in the glandular stomach. This is indicative of a reactive, point of contact genotoxin which is not available for systemic exposure.

Link to relevant study records

Reference

Reference 1

Endpoint:

in vivo mammalian cell study: DNA damage and/or repair

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 489 (In vivo Mammalian Alkaline Comet Assay)

Deviations:

not specified

GLP compliance:

not specified

Type of assay:

mammalian comet assay

Species:

mouse

Strain:

B6C3F1

Sex:

male

Details on test animals or test system and environmental conditions:

Age 8-10 weeks
body weight 20-35 g
supplier Taconic NIEHS colony
Food: Purina Certified 5002 Pelleted Diet, ad libitum

Housed singly
Temperature 18-26 celsius
relative humidity 30%
Light period 600-1800

Route of administration:

oral: gavage

Vehicle:

0.9% w/v saline

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: prepared in 0.9 % saline
at 2.5mg/ml, 5 mg/ml and 10 mg/ml

Frequency of treatment:

single dose

Post exposure period:

72 hours

Dose / conc.:

25 mg/kg bw/day

Dose / conc.:

50 mg/kg bw/day

Dose / conc.:

100 mg/kg bw/day

No. of animals per sex per dose:

5 (five)

Control animals:

yes, concurrent vehicle

Positive control(s):

ethyl methanesulfonate 150 mg/kg/day

Tissues and cell types examined:

blood; colon; liver,left lobe; glandular stomach

Details of tissue and slide preparation:

tissues were lysed overnight and DNA samples prepared on Trevigen slides in 0.5% agarose. The DNA was unwound for 20 minutes at a pH of 13.2. Electrophoresis : 25V, 300 mA starting current at 28% relative humidity/21C temperature in a GTX5 COMET electophoresis system. DNA stained with SYBR gold stain.

Evaluation criteria:

COMET tails assessed using Perceptive Instruments software verison 4.11

Sex:

male

Genotoxicity:

positive

Toxicity:

not examined

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

valid

Remarks on result:

other: glandular stomach

Sex:

male

Genotoxicity:

negative

Toxicity:

not examined

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

valid

Remarks on result:

other: blood, colon, left liver lobe

Additional information on results:

All assessments were completed based on COMET tail length.

Mean COMET tail lengths in tested tissues

Dose Level	COMET tail length mean	COMET tail length (SEM)	pairwise P value	Organ	Pairwise Test
Vehicle Control	26.52	0.85		Blood
Low Dose	29.45	0.82	0.154	Blood	Williams
Mid Dose	26.21	0.93	0.186	Blood	Williams
High Dose	30.36	0.91	0.004	Blood	Williams
Positive Control	58.20	2.31	0.005	Blood	Dunn
Vehicle Control	44.88	2.68		Colon
Low Dose	46.94	3.23	0.384	Colon	Williams
Mid Dose	45.36	2.88	0.452	Colon	Williams
High Dose	50.18	3.59	0.164	Colon	Williams
Positive Control	65.58	5.11	0.004	Colon	Williams
Vehicle Control	56.12	2.26		Liver
Low Dose	60.19	3.00	0.781	Liver	Williams
Mid Dose	43.21	2.13	0.857	Liver	Williams
High Dose	57.18	3.01	0.493	Liver	Williams
Positive Control	71.95	2.25	0.001	Liver	Williams
Vehicle Control	42.69	2.03		Stomach
Low Dose	64.31	4.92	0.038	Stomach	Williams
Mid Dose	66.35	10.15	0.032	Stomach	Williams
High Dose	75.73	11.37	0.006	Stomach	Williams
Positive Control	84.25	6.99	0.000	Stomach	Williams

Test for linear trend:

Colon	LinearTrend	0.138105
Blood	LinearTrend	0.030227
Stomach	LinearTrend	0.007354
Liver	LinearTrend	0.629107

Conclusions:

A significant increase in COMET tail length was observed in the glandular stomachs of male mice treated by oral gavage with the substance; the increase had a significant linear trend. Significant dose dependant increases in tail length were not observed in the other tissues tested. The investigator concluded that the substance was genotoxic in the glandular stomach, indicating a point of contact, rather than systemic genotoxin.

Reference 2

Endpoint:

in vivo mammalian cell study: DNA damage and/or repair

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

GLP compliance:

yes

Type of assay:

mammalian erythrocyte micronucleus test

Species:

mouse

Strain:

CB6F1

Sex:

male

Details on test animals or test system and environmental conditions:

Age 8-10 weeks
body weight 20-35 g
supplier Taconic NIEHS colony
Food: Purina Certified 5002 Pelleted Diet, ad libitum

Housed singly
Temperature 18-26 celsius
relative humidity 30%
Light period 600-1800

Route of administration:

oral: gavage

Vehicle:

0.9% saline

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: prepared in 0.9 % saline
at 2.5mg/ml, 5 mg/ml and 10 mg/ml

Duration of treatment / exposure:

72 hours

Frequency of treatment:

single

Dose / conc.:

25 mg/kg bw/day (actual dose received)

Dose / conc.:

50 mg/kg bw/day (actual dose received)

Dose / conc.:

100 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

5 (five)

Control animals:

yes, concurrent vehicle

Positive control(s):

150 mg/kg ethyl methane sulfonate

Tissues and cell types examined:

bone marrow erythrocytes

Sex:

male

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

valid

Blood
Start Date	Sample Collection Time	Sex	Methodology Used	Route	Dosing Regimen
23/09/2008	28 hour	Male	Flow Cytometry	Gavage	Gavage x 4, 4 day
		Dose	Animal	Polychromatic Erythrocytes				Normochromatic Erythrocytes
		(mg/kg)	Number	Trend P: 0.9057418632200795				Trend P: 0.5630751976468024
				No. Examined	Total MN Cells	Percent PCE	MN Cells	No. Examined	Total MN Cells	Percent NCE	MN Cells
							per 1000				per 1000
Vehicle Control	Saline	0	1	20000	62	1.4	3.1	1428487	2095	98.6	1.5
		0	2	20000	57	1.1	2.9	1724165	2304	98.9	1.3
		0	3	20000	49	1.3	2.5	1522067	2273	98.7	1.5
		0	4	20000	51	1.4	2.6	1389274	2125	98.6	1.5
		0	5	20000	54	1.2	2.7	1669659	2434	98.8	1.5
Average ± SEM						1.285 ± 0.053	2.730 ± 0.115			0.000 ± 0.000	1.457 ± 0.033
Test Chemical	1,3-Dichloro-2-propanol	25	10	20000	44	1.6	2.2	1201091	1817	98.4	1.5
		25	6	20000	52	1.4	2.6	1437928	2142	98.6	1.5
		25	7	20000	48	1.3	2.4	1576941	2422	98.7	1.5
		25	8	20000	40	1.3	2	1543362	2215	98.7	1.4
		25	9	20000	62	1.3	3.1	1482301	2267	98.7	1.5
Average ± SEM						1.375 ± 0.069	2.460 ± 0.189			0.000 ± 0.000	1.501 ± 0.018
Pairwise P							0.817				0.349
Test Chemical	1,3-Dichloro-2-propanol	50	11	20000	45	1.3	2.3	1549017	2174	98.7	1.4
		50	12	20623	41	1.2	2	1662551	2475	98.8	1.5
		50	13	20000	38	1.1	1.9	1769073	2455	98.9	1.4
		50	14	20000	49	1.3	2.5	1475861	2145	98.7	1.5
		50	15	20000	62	1.6	3.1	1222994	1825	98.4	1.5
Average ± SEM						1.313 ± 0.082	2.338 ± 0.214			0.000 ± 0.000	1.445 ± 0.021
Pairwise P							0.885				0.415
Test Chemical	1,3-Dichloro-2-propanol	100	16	20000	56	1	2.8	1903261	2781	99	1.5
		100	17	20000	37	1.1	1.9	1757290	2554	98.9	1.5
		100	18	20000	46	0.8	2.3	2494753	3909	99.2	1.6
		100	19	20000	61	0.8	3.1	2391448	3396	99.2	1.4
		100	20	20000	33	0.9	1.7	2220912	3186	99.1	1.4
Average ± SEM						0.936 ± 0.063	2.330 ± 0.268			0.000 ± 0.000	1.467 ± 0.026
Pairwise P							0.909				0.443
Positive Control	Ethyl Methane Sulfonate	150	21	20000	177	1	8.9	2020359	3163	99	1.6
		150	22	20000	147	1.2	7.4	1637685	2642	98.8	1.6
		150	23	20000	235	1.1	11.8	1806498	3315	98.9	1.8
		150	24	20000	234	1.2	11.7	1718822	2967	98.8	1.7
		150	25	20000	203	1	10.2	1947659	3185	99	1.6
Average ± SEM						1.090 ± 0.042	9.960 ± 0.846			0.000 ± 0.000	1.675 ± 0.048
Pairwise P							0.005				0.003

Conclusions:

1,3-dichloropropan-2-ol did not induce any micronuclei in the femur bone marrow of male B6C3F1 mice at 25-100 mg/kg bw when examined using the OECD 474 test guideline

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (positive)

Justification for classification or non-classification

The weight of evidence presented by the results of the Ames studies (positive in a number of strains of S.typhimurium) indicate that the substance does have mutagenic potential. However, the negative somatic mouse micronucleus study and negative findings in the somatic tissues in the mouse COMET assay indictate that there is negiligible concern that the substance is a Germ Cell mutagen. The positive findings in the mouse glandular stomach following oral dosing indicate that there is a point mutagen potential; in conjunction with the known carcinogenic properties of the substance, it is therefore recommended that the substance be classified as Germ Cell Mutagen Category 2 in accordance with the criteria set out in 1272/2008/EC.