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Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Gene mutation in vitro:

Ames test:

The test chemical 2,3-bis[(4-methylbenzoyl)oxy]succinic acid did not induce gene mutation in Salmonella typhimurium strains and E. coli WP2 in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.

Link to relevant study records
Reference
Endpoint:
in vitro gene mutation study in bacteria
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Remarks:
experimental data of read across substances
Justification for type of information:
Data for the target chemical is summarized based on the structurally similar read across chemicals
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Qualifier:
according to
Guideline:
other: Refer below principle
Principles of method if other than guideline:
WoE derived based on the experimental data from structurally and functionally similar read across chemicals
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay
Target gene:
1. Histidne for Salmonella typhimurium strains and tryptophan for E.coli strains
2. Histidine
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Remarks:
1
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
E. coli WP2 uvr A pKM 101
Remarks:
1
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
S. typhimurium, other: TA92, TA1535, TA100, TA1537, TA94 and TA98
Remarks:
2
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
not specified
Cytokinesis block (if used):
No data
Metabolic activation:
with and without
Metabolic activation system:
S 9 is a 9000 × g centrifugal supernatant fraction of liver homogenate prepared by administering phenobarbital and 5,6-benzoflavone in combination with 7-week-old male SD rats
Test concentrations with justification for top dose:
1.
-S9: 0, 313, 625, 1250, 2500 or 5000 µg/plate
+S9: 0, 156, 313, 625, 1250, 2500 or 5000 µg/plate

2. 6 different concentrations were used; 10.0 mg/plate was the maximum concentration
Vehicle / solvent:
1.
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test chemical was soluble in DMSO

2.
- Vehicle(s)/solvent(s) used: Phosphate buffer
- Justification for choice of solvent/vehicle: The test chemical was soluble in phosphate buffer
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
N-ethyl-N-nitro-N-nitrosoguanidine
other: 2-aminoanthracene and 2- (2-furyl) -3- (5-nitro-2-furyl) acrylamide
Remarks:
1
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
Phosphate buffer
True negative controls:
not specified
Positive controls:
not specified
Positive control substance:
not specified
Remarks:
2
Details on test system and experimental conditions:
1. METHOD OF APPLICATION: Preincubation

DURATION
- Preincubation period: 20 mins
- Exposure duration: 48 hrs
- Expression time (cells in growth medium): 48 hrs
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data

SELECTION AGENT (mutation assays): No data
SPINDLE INHIBITOR (cytogenetic assays): No data
STAIN (for cytogenetic assays): No data

NUMBER OF REPLICATIONS: Three plates were used for each dose in this test, and twice to confirm reproducibility

NUMBER OF CELLS EVALUATED: No data

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data

OTHER EXAMINATIONS: No data
- Determination of polyploidy: No data
- Determination of endoreplication: No data
- Other: No data

OTHER: No data

2. METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 mins
- Exposure duration: 48 hrs
- Expression time (cells in growth medium): 48 hrs
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data

SELECTION AGENT (mutation assays): No data
SPINDLE INHIBITOR (cytogenetic assays): No data
STAIN (for cytogenetic assays): No data

NUMBER OF REPLICATIONS: Duplicate

NUMBER OF CELLS EVALUATED: No data

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data

OTHER EXAMINATIONS:
- Determination of polyploidy: No data
- Determination of endoreplication: No data
- Other: No data

OTHER: No data
Rationale for test conditions:
1. No data
Evaluation criteria:
1. Regardless of the presence or absence of S9 mix in any of the test strains, the number of reversed mutantcolonies (mean value) increased with the increase in the dose of the test substance to more than twice the negative (solvent) control value, When reproducibility was observed, the test substance was determined to be mutagenic (positive).

2. The result was considered positive if the number of colonies found was twice the number in the control (exposed to the appropriate solvent or untreated).
Statistics:
1. No statistical method was used for judging the test results
2. No data
Species / strain:
S. typhimurium, other: TA98, TA100, TA1535, TA1537
Remarks:
1
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
E. coli WP2 uvr A pKM 101
Remarks:
1
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
S. typhimurium, other: TA92, TA1535, TA100, TA1537, TA94 and TA98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Additional information on results:
1.
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: No data
- Effects of osmolality: No data
- Evaporation from medium: No data
- Water solubility: No data
- Precipitation: No data
- Other confounding effects: No data

RANGE-FINDING/SCREENING STUDIES: Preliminary test was carried out at 7 doses of 5000, 1250, 313, 78.1, 19.5, 4.88 and 1.22 μg / plate using
Salmonella typhimurium TA 100, TA 1535, TA 98, TA 1537 and Escherichia coli WP 2 uvr A / pKM 101, no increase in the number of revertant colonies was observed in any of the strains. In all the strains coexisting withS9 mix, antibacterial activity was observed at 5000 μg / plate.

COMPARISON WITH HISTORICAL CONTROL DATA: No data

ADDITIONAL INFORMATION ON CYTOTOXICITY: No data

2. ADDITIONAL INFORMATION ON CYTOTOXICITY: The maximum dose for negative results represents the highest non-cytotoxic dose used in the experiment
Remarks on result:
other: No mutagenic potential
Conclusions:
The test chemical 2,3-bis[(4-methylbenzoyl)oxy]succinic acid did not induce gene mutation in Salmonella typhimurium strains and E. coli WP2 in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
Executive summary:

Data available for the test chemicals was reviewed to determine the mutagenic nature of 2,3-bis[(4-methylbenzoyl)oxy]succinic acid. The studies are as mentioned below:

Ames assay was performed to determine the mutagenic nature of the test chemical. The study was performed by the preincubation protocol using Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and E. coli WP2 uvrA / pKM101 in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in DMSO and used at dose level of 0, 313, 625, 1250, 2500 or 5000µg/plate in the absence of S9 and 0, 156, 313, 625, 1250, 2500 or 5000µg/plate in the presence of S9. Concurrent solvent and positive control plates were also included in the study. Based on the observations made, the test chemical did not induce gene mutation in Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and E. coli WP2 uvrA / pKM101 in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.

In another study, gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The study was performed using S. typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 with and without S9 metabolic activation system. The test was performed as per the preincubation assay at six different concentrations with 10.0 mg/plate being the maximum concentration. The chemical was dissolved in phosphate buffer. Preincubation was performed for 20 mins and the exposure duration was for 48 hrs. The result was considered positive if the number of colonies found was twice the number in the control (exposed to the appropriate solvent or untreated). The test chemical did not induce a doubling of revertant colonies over the control using S. typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 in the presence and absence of S9 metabolic activation system and hence the chemical is not likely to classify as a gene mutant in vitro.

Based on the observations made, the test chemical 2,3-bis[(4-methylbenzoyl)oxy]succinic acid did not induce gene mutation in Salmonella typhimurium strains and E. coli WP2 in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Gene mutation in vitro:

Ames test:

Data available for the test chemicals was reviewed to determine the mutagenic nature of 2,3-bis[(4-methylbenzoyl)oxy]succinic acid (CAS no 32634 -68 -7). The studies are as mentioned below:

Ames assay was performed to determine the mutagenic nature of the test chemical. The study was performed by the preincubation protocol using Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and E. coli WP2 uvrA / pKM101 in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in DMSO and used at dose level of 0, 313, 625, 1250, 2500 or 5000µg/plate in the absence of S9 and 0, 156, 313, 625, 1250, 2500 or 5000µg/plate in the presence of S9. Concurrent solvent and positive control plates were also included in the study. Based on the observations made, the test chemical did not induce gene mutation in Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and E. coli WP2 uvrA / pKM101 in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.

In another study, gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The study was performed using S. typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 with and without S9 metabolic activation system. The test was performed as per the preincubation assay at six different concentrations with 10.0 mg/plate being the maximum concentration. The chemical was dissolved in phosphate buffer. Preincubation was performed for 20 mins and the exposure duration was for 48 hrs. The result was considered positive if the number of colonies found was twice the number in the control (exposed to the appropriate solvent or untreated). The test chemical did not induce a doubling of revertant colonies over the control using S. typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 in the presence and absence of S9 metabolic activation system and hence the chemical is not likely to classify as a gene mutant in vitro.

Based on the observations made, the test chemical 2,3-bis[(4-methylbenzoyl)oxy]succinic acid (CAS no 32634 -68 -7) did not induce gene mutation in Salmonella typhimurium strains and E. coli WP2 in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.

Justification for classification or non-classification

Based on the observations made, 2,3-bis[(4-methylbenzoyl)oxy]succinic acid (CAS no 32634 -68 -7) does nor exhibit gene mutation in vitro. Hence the test chemical is not likely to classify as a gene mutant in vitro.