D-Mannitol, 1,4:3,6-dianhydro-, 2,5-bis[3-methoxy-4-[[4-[(1-oxo-2-propen-1-yl)oxy]benzoyl]oxy]benzoate]

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:

2008-06-18 to 2008-08-08

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD Guideline study report

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of (3R,3aS,6R,6aS)-6-[(4-{[4-(acryloyloxy)benzoyl]oxy}-3-methoxybenzoyl)oxy]hexahydrofuro[3,2-b]furane-3-yl 4-{[4-(acryloyloxy)benzoyl]oxy}-3-methoxybenzoate and (3R,3aR,6R,6aR)-6-(4-(4-(acryloyloxy)benzoyloxy)-3-methoxybenzoyloxy)hexahydrofuro[3,2-b]furan-3-yl 4-(4-(4-(acryloyloxy)benzoyloxy)benzoyloxy)-3-methoxybenzoate + Isomers and (3R,3aR,6R,6aR)-6-(4-(4-(4-(acryloyloxy)benzoyloxy)-3- methoxybenzoyloxy)-3-methoxybenzoyloxy)hexahydrofuro[3,2-b]furan-3-yl 4-(4-(acryloyloxy)benzoyloxy)-3-methoxybenzoate + Isomers

IUPAC Name:

Reaction mass of (3R,3aS,6R,6aS)-6-[(4-{[4-(acryloyloxy)benzoyl]oxy}-3-methoxybenzoyl)oxy]hexahydrofuro[3,2-b]furane-3-yl 4-{[4-(acryloyloxy)benzoyl]oxy}-3-methoxybenzoate and (3R,3aR,6R,6aR)-6-(4-(4-(acryloyloxy)benzoyloxy)-3-methoxybenzoyloxy)hexahydrofuro[3,2-b]furan-3-yl 4-(4-(4-(acryloyloxy)benzoyloxy)benzoyloxy)-3-methoxybenzoate + Isomers and (3R,3aR,6R,6aR)-6-(4-(4-(4-(acryloyloxy)benzoyloxy)-3- methoxybenzoyloxy)-3-methoxybenzoyloxy)hexahydrofuro[3,2-b]furan-3-yl 4-(4-(acryloyloxy)benzoyloxy)-3-methoxybenzoate + Isomers

Details on test material:

Name: 2MeO3-Im4KA2
Batch no.: AC425/428/429/430
Chemical name: (3R,3aS, 6R,6aS)-6-[(4-{[4-(acryloyloxy) benzoyl]oxy}-3-methoxybenzoyl)oxy] hexahydrofuro[3,2-b]furan-3-yl 4-{[4-
(acryloyloxy)benzoyl]oxy}-3- methoxybenzoate
Purpose: industrial chemical
Colour: beige
Physical state: powder
Stability in solution
or suspension: probably stable
Special procedures
for dose preparation: prepare freshly
Storage: at room temperature, protected from light

Method

Target gene:

not applicable

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Details on mammalian cell type (if applicable):

TA 98: his D 3052, rfa-; uvrB-; R-factor
TA 100: his G 46, rfa-; uvrB-; R-factor
TA 1535: his G 46, rfa-; uvrB-
TA 1537: his C 3076, rfa-; uvrB-

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 102

Details on mammalian cell type (if applicable):

TA 102: his G 428 (pAQ1), rfa-; uvrB-; R-factor

Metabolic activation:

with and without

Metabolic activation system:

S9

Test concentrations with justification for top dose:

Pretest: 3.16, 10, 31.6, 100, 316, 1000, 2500 and 5000 µg/plate with/without activation
Main experiment 1 : 31.6, 100, 316, 1000, 2500 and 5000 µg/plate without activation (TA 98, TA 100, TA 1535, TA 1537)
Main experiment 1 : 1, 3.16, 10.0, 31.6, 100, 316 and 1000 µg/plate without activation (TA 102)
Main experiment 1: 31.6, 100, 316, 1000, 2500 and 5000 µg/plate with activation
Main experiment 2: 10, 31.6, 100, 316, 1000, 2500 and 5000 µg/plate without activation (TA 98, TA 100)
Main experiment 2: 1, 3.15, 10, 31.6, 100, 316 and 1000 µg/plate without activation (TA 102, TA 1535, TA 1537)
Main experiment 2: 10, 31.6, 100, 316, 1000, 2500 and 5000 µg/plate with activation

Vehicle / solvent:

DMSO

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

For TA 100, TA 1535 without S9

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive controls:

yes

Positive control substance:

other: 4-Nitro-o-phenylene-diamine

Remarks:

For TA 98, TA 1537 without S9

Controls 3

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracene

Remarks:

all strains with S9

Controls 4

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

For TA 102 without S9

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation, 1st main experiment); preincubation (2nd main experiment)


DURATION
- Preincubation period: 60 min
- Exposure duration: 48 h


NUMBER OF REPLICATIONS: 3

Evaluation criteria:

The test is considered acceptable if the bacteria demonstrate their typical responses to ampicil-lin (TA 98, Ta 100, TA 102); the control plates with and without S9 are within the following ranges (mean values of the spontaneous reversion frequency are within the historical control data range: TA 98 18-54 (-S9), 18-71 (+S9); TA 100 75-167 (-S9), 81-168 (+S9); TA 102 165-391 (-S9), 163-594 (+S9); TA 1535 5-29 (-S9), 6-31 (+S9); TA 1537 5-30 (-S9), 6-36 (+S9)); corresponding background growth on negative control, solvent control and test plates is ob-served; the positive controls show a distinct enhancement of revertant rates over the control plate. A biologically relevant increase with or without metabolic activation is
- if in tester strains TA100 and TA 102 the number of reversions is at least twice as high
- if in the other tester strains the number of reversions is at least three times higher
than the reversion rate of the solvent control.

Statistics:

not reported

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

>= 2500 µg/plate (experiment I without S9), >= 316 µg/plate (experiment II without S9),

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

>= 1000 µg/plate (experiment I without S9), >= 31.6 µg/plate (experiment II without S9), >= 2500 µg/plate (experiment II with S9),

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

>= 1000 µg/plate (experiment I without S9), >= 31 µg/plate (experiment II without S9), >= 2500 µg/plate (experiment II with S9),

Vehicle controls validity:

valid

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

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

>= 316 µg/plate (experiment I without S9), >= 5000 µg/plate (experiment I with S9), >= 31.6 µg/plate (experiment II without S9),

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

>= 100 µg/plate (experiment I without S9), >= 100 µg/plate (experiment II without S9),

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: none
- Effects of osmolality: none
- Evaporation from medium: none
- Water solubility: none
- Precipitation: yes, 1st experiment > = 316 µg/plate (without S9), > = 2500 µg/plate (with S9), 2nd experiment > = 31.6 µg/plate (without S9), >= 1000 µg/plate (with S9)


RANGE-FINDING/SCREENING STUDIES: yes

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

The substance did not induce any mutagenic effect in the Ames test with and without metabolic activation under the conditions used.

Executive summary:

The test item 2MeO3-Im4K2 was investigated for its potential to induce gene mutations according to the plate incorporation test (experiment I) and the pre-incubation test (experiment II) using Salmonella typhimurium strains TA 98, TA 100, TA 102, TA 1535 and TA 1537.

In two independent experiments several concentrations of the test item were used. Each assay was conducted with and without metabolic activation. The concentrations, including the controls, were tested in triplicate.

The following concentrations were used:

Pretest: 3.16, 10, 31.6, 100, 316, 1000, 2500 and 5000 µg/plate with/without activation

Main experiment 1 : 31.6, 100, 316, 1000, 2500 and 5000 µg/plate without activation (TA 98, TA 100, TA 1535, TA 1537)

Main experiment 1 : 1, 3.16, 10.0, 31.6, 100, 316 and 1000 µg/plate without activation (TA 102)

Main experiment 1: 31.6, 100, 316, 1000, 2500 and 5000 µg/plate with activation

Main experiment 2: 10, 31.6, 100, 316, 1000, 2500 and 5000 µg/plate without activation (TA 98, TA 100)

Main experiment 2: 1, 3.15, 10, 31.6, 100, 316 and 1000 µg/plate without activation (TA 102, TA 1535, TA 1537)

Main experiment 2: 10, 31.6, 100, 316, 1000, 2500 and 5000 µg/plate with activation

Precipitation of the test item was observed in all tester strains used in experiments I and II (with and without metabolic activation). In experiment I precipitation of the test item was found at doses of 316 µg/plate and higher (without metabolic activation) and at doses of 2500 µg/plate (with metabolic activation), in experiment II precipitation of the test item was found at doses of 31.6 µg/plate and higher (without metabolic activation) and at a doses of 1000 µg/plate and higher(with metabolic activation).

Toxic effects of the test item were noted in all tester strains evaluated in experiments I and II. In experiment I toxic effects of the test item were observed in tester strain TA 98 at doses of 2500 µg/plate and higher (without metabolic activation). In tester strains TA 100 and TA 1535 toxic effects were noted at doses of 1000 µg/plate and higher. In the tester strain TA 1537 toxic effects were observed at doses of 316 µg/plate without metabolic activation and at a does of 5000 µg/plate (with metabolic activation). In tester starin TA 102 toxic effects of the test item were noted at doses of 100 µg/plate and higher (without metabolic activation.). The reduction in the number of revertants down to a mutation factor of 0.5 found in tester strain TA 1535 at a does of 100 µg/plate (without metabolic activation) was regarded as not biologically relevant due to lack of a dose-response relationship.

In experiment II toxic effects of the test item were noted in tester strain TA 98 at doses of 316 µg/plate and higher (without metabolic activation). In tester strains TA 100 and TA 1535 toxic effects of the test item were observed at doses of 31.6 µg/plate and higher (without metabolic activation) and at doses of 2500 mg/plate and higher (with metabolic activation). In tester strain TA 1537 toxic effects of the test item were seen at doses of 31.6 µg/plate and higher (without metabolic activation). In tester starin TA 102 toxic effects of the test item were noted at doses of 100 mg/plate and higher (without metabolic activation).

No biologically relevant increases in revertant colony numbers of any of the five tester strains were observed following treatment with 2MeO3-Im4K2 at any concentration level, neither in the presence nor absence of metabolic activation in experiments I and II.

The reference mutagens induced a distinct increase of revertant colonies indicating the validity of the experiments.

In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, 2MeO3-Im4K2 did not cause gene mutations by base pair changes or frameshifts in the genome of the tester strains used. Therefore, 2MeO3-Im4K2 is considered to be non-mutagenic in this bacterial reverse mutation assay according to OECD guideline 471.