OO-tert-butyl O-(2-ethylhexyl) peroxycarbonate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1985

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study but without raw data.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1985

Report date:

1985

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

1983

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

OO-tert-butyl O-(2-ethylhexyl) peroxycarbonate

EC Number:

252-029-5

EC Name:

OO-tert-butyl O-(2-ethylhexyl) peroxycarbonate

Cas Number:

34443-12-4

Molecular formula:

C13H26O4

IUPAC Name:

3-({[(tert-butylperoxy)carbonyl]oxy}methyl)heptane

Test material form:

other: liquid

Method

Species / strain

Species / strain / cell type:

other: TA 1535, 1537, 1538, 98 and 100

Details on mammalian cell type (if applicable):

strains regularly checked for their histidine requirement, ampicillin resistance (TA98 and TA100) and the frequency of spontaneous revertants

Metabolic activation:

with and without

Metabolic activation system:

S9 (following intraperitoneal injection of adult male Wistar rats of Aroclor 1524 (500 mg/kg bw) in corn oil, rats were killed by decapitation, livers were removed and the supernactant was extracted.)

Test concentrations with justification for top dose:

0, 100, 333, 1000, 3333, 5000 µg/plate

Vehicle / solvent:

DMSO excepting for positive controls: DMS for TA100, TA98 and TA1538 without activation, and for all strains with activation.
Water for TA1535 and TA1537.

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: Without metabolic activation: TA 1535: sodium azide; TA 100: methylmethanesulfonate; TA 98 and TA 1538: 4-nitro-o-phenylene-diamine; TA1537: 9-aminoacridine /with metabolic activation: 2-aminoanthracene for all strains

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

EXPOSURE DURATION: 48 hours at 37 °C

NUMBER OF REPLICATIONS: 3

Results and discussion

Test results

Test results 1

Species / strain:

other: all strains

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Species / strain:

other: TA1535, TA1537, TA1538

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

from 1000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 3

Species / strain:

other: TA98 and TA100

Metabolic activation:

with

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

other: yes, but only for TA98 and from 3333 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Any other information on results incl. tables

	Mean number of revertant (His+), colonies /3 replicate plates WITHOUT S9
Dose (µg/plate)	TA1535	TA1537	TA1538	TA98	TA100
100	7+-3	7+-4	5+-1	30+-5	125+-13
333	9+-3	5+-1	4+-1	25+-5	99+-25
1000	9+-2	6+-3	7+-3	25+-4	85+-25
3333	9+-1	9+-4	7+-3	29+-2	92+-10
5000	11+-7	10+-2	7+-3	33+-7	87+-15
Solvent control	7+-3	8+-3	7+-2	17+-2	85+-26
Positive control	320+-22	1282+-138	1163+-32	860+-141	775+-33
	Mean number of revertant (His+), colonies /3 replicate plates WITH S9
Dose (µg/plate)	TA1535	TA1537	TA1538	TA98	TA100
100	10+-0	14+-2	16+-3	54+-7	168+-10
333	8+-1 (b)	20+-7(b)	17+-1(b)	90+-10	223+-23(b)
1000	0(d)	0(d)	0(d)	26+-3	334+-30(b)
3333**	0(d)	0(d)	0(d)	9*(d)	354+-211(c )
5000**	0(d)	0(d)	0(d)	14*(d)	316+-23(d)
Solvent control	8+-1	10+-1	16+-2	32+-4	91+-2
Positive control	170+-17	71+-7	1106+-22	992+-80	1787+-140
b)	slightly reduced background bacterial lawn
c)	moderately reduced background bacterial lawn
d)	background bacterial lawn extremely reduced or absent
*	2 plates with no bacterial growth

Applicant's summary and conclusion

Conclusions:

TBEC shown mutagenic activity in the bacterial reverse mutation test with Salmonella typhimurium.

Executive summary:

The potential of TBEC to induce reverse mutation in Salmonella typhimurium (strains: TA 1535, TA 1537, TA1538, TA 98, TA 100) was evaluated in accordance with the international guidelines (OECD 471, 1986) and in compliance with the Principles of Good Laboratory Practice.

TBEC was tested in two independent experiments, with and without a metabolic activation system, according to direct plate incorporation method, without and with S9.

 

Bacterias were exposed to TBEC at six dose-levels (three plates/dose-level) selected from a preliminary toxicity test: 0, 100, 333, 1000, 3333, 5000 µg/plate. After 48 hours of incubation at 37°C, the revertant colonies were scored.

 

No cytotoxicity and no noteworth increase in the number of revertants was noted without S9.

With S9, Cytotoxicity was clearly observed from 1000 µg/plate for strains TA1535, TA1537, TA1538.

TBEC produced dose-related increases in the number of revertants of testers strains TA98 and TA100 in the presence of S9.

In conclusion, under these experimental conditions, TBEC shown mutagenic activity in the bacterial reverse mutation test with Salmonella typhimurium.