2,3-epoxypropyl phenyl ether

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:

Not reported

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Although this study was not conducted in accordance with current international guidelines or GLP, in accordance with REACH Annex XI, Section 1.1.2, the study was conducted under sound scientific principles of the bacterial reverse mutation test and adequate documentation of the study is provided. The study is therefore considered adequate for fulfilling this endpoint and for risk assessment purposes.

Data source

Reference

Reference Type:

publication

Title:

No information

Author:

Elliott J. Greene

Year:

1979

Bibliographic source:

Greene, E.J., Friedman, M.A., Sherrod, J.A., Salerno, A.J. (1979) In-vitro mutagenicity and cell transformation screening of phenylglycidyl ether. Mutation Research, 67:9-19.

Materials and methods

Principles of method if other than guideline:

The plate test described by Ames et al. was used:
Ames, B.N., McCann, J. and Yamasaki, E. (1975) Methods for detecting carcinogens and mutagens with the salmonella/mammalian-microsome mutagenicity test. Mutation Research, 31:347-364.

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2,3-epoxypropyl phenyl ether

EC Number:

204-557-2

EC Name:

2,3-epoxypropyl phenyl ether

Cas Number:

122-60-1

Molecular formula:

C9H10O2

IUPAC Name:

2-(phenoxymethyl)oxirane

Details on test material:

Phenylglycidyl ether (>99 % purity) was obtained from Shell Chemical Co. (Houston, TX).

Method

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:

S. typhimurium TA 1538

Metabolic activation:

with and without

Metabolic activation system:

Rat S9

Details on test system and experimental conditions:

Bacterial Media: Nutrient agar was prepared from powdered nutrient broth (BBL, Cockeysville, MD) supplemented with 1.5 % agar (DIFCO, Detroit, MI). Minimal agar and top agar were prepared using DIFCO agar. Autoclaved nutrient and minimal agar were dispensed into sterile 100 x 15 mm petri dishes (Fisher Scientific, Pittsburgh, PA), 20 ml per plate. Top agar was stored in 2 ml volumes in 13 x 100 mm sterile, screw cap plastic tubes (Falcon, Oxnard, CA).

Rat S9 was prepared from CDF-344 rats which were obtained from Charles River Laboratories (Wilmington, MA), housed 4-6 cage, and given Purina Rat Chow and water ad libitum. Rats were injected with 75 mg/kg Aroclor 1254 5 days prior to use. Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and TA1538 were obtained from Dr Bruce Ames (University of California, Berkeley). Preparation of stock cultures and other manipulations were carried out according to Ames et al. (1975).

The plate test described by Ames et al. (1975) was used. The appropriate bacterial strain was grown overnight at 37 degrees celsius in nutrient broth (BBL). The cultures were then centrifuged, washed 2-3 times in sterile, physiological saline, and resuspended to the original volume in saline. 0.1 ml of this suspension was mixed with 2-2.5 ml top agar and 0.5 ml rat S9 mix, prepared according to Ames et al. (1975). For the spot test, this mixture was then poured onto a minimal agar plate. Otherwise, the chemical was also added in 25 μL volume, and the mixture was poured directly onto the agar overlay. After the overlay had hardened, the plates were incubated at 37 degrees celsius for 2 days. Toxicity was determined by repeating the above procedure, but using 10^-5, 10^-6 and 10^-7 dilutions of bacteria, pouring onto the nutrient agar plates, which were incubated at 37 degrees celsius for 24 hours.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

positive

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

positive

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

The bacterial mutagencitiy of PGE was tested first in the spot test. The results (not presented) demonstrated the classic pattern of a zone of inhibition, then high numbers of revertants tailing off to background with increasing radius, in S. typhimurium strains TA1535 and TA100 with and without metabolic activation. These tests were repeated using the plate-incorporation technique. The results (Table 1) reaffirmed the spot test data. Phenylglycidyl ether was mutagenic in a dose-dependent fashion in strains TA1535 and TA100. Activating medium was not required for mutagenic activity.

Any other information on results incl. tables

Table 1. Bacterial mutagenicity of PGE

Chemical	Amount per plate (μg)	S9^b	Revertants per plate^a
			TA98	TA100	TA1535	TA1537	TA1538
Negative control	-	-	12	62	10	8	6
Negative control	-	+	20	68	6	8	10
Positive control^c	-	-	784	259	29	203	721
Positive control^d	-	+	204	1380	216	52	216
PGE	0.5	-	16	51	13	4	11
	50	-	10	72	28	7	8
	50	-	9	204	146	7	10
	500	-	10	1158	590	15	12
	0.5	+	20	69	9	6	21
	5	+	12	73	8	5	14
	50	+	18	136	26	9	14
	500	+	19	1114	487	14	23

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
positive with and without metabolic activation

Concentration dependent mutagenicity was demonstrated in S. typhimurium strains TA1535 and TA100 with and without metabolic activation, but not in strains TA98, TA1537 or TA1538.