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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
Further information is included under 'Attached justification' in IUCLID section 13 and 'Cross-reference'.
Cross-reference
Reason / purpose:
read-across: supporting information
Reference
Endpoint:
in vitro gene mutation study in bacteria
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
Further information is included under 'Attached justification' in IUCLID section 13 and 'Cross-reference'.
Reason / purpose:
read-across source
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source of test material:
Nisshin Oillo Group, Ltd., Tokyo, Japan
- Composition: Caprylic acid (9.7%), capric acid (3.3%), palmitic acid (3.8%), stearic acid (1.7%), oleic acid (51.2%), linoleic acid (18.4%), linolenic acid (9.0%) and other fatty acids (2.9%)
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
Type and composition of metabolic activation system:
- concentration or volume of S9 mix and S9 in the final culture medium: 0.5 ml
Test concentrations with justification for top dose:
0, 313, 625, 1250, 2500 and 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: no data - vehicle control used
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
benzo(a)pyrene
other: NaN3, AF-2, ICR-191
Details on test system and experimental conditions:
NUMBER OF REPLICATIONS:
- Number of cultures per concentration: triplicate

METHOD OF TREATMENT/ EXPOSURE:
- Test substance added in agar (plate incorporation)

TREATMENT AND HARVEST SCHEDULE:
- Preincubation period, if applicable: 20 minutes
Rationale for test conditions:
A preliminary study was conducted.
Evaluation criteria:
Not stated.
Statistics:
Not stated.
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
True negative controls validity:
not examined
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not determined
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
True negative controls validity:
not examined
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not determined
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
True negative controls validity:
not examined
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not determined
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
True negative controls validity:
not examined
Positive controls validity:
valid
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not determined
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
True negative controls validity:
not examined
Positive controls validity:
valid

  Dose (µg/plate)                 
 Strain 313  625  1250  2500  5000 
 TA98 (-) 24 25  17  17  22  24 
 TA100 (-) 129  140  123  136  122  136 
 TA1535 (-) 21  29  19  23  23  20 
 TA1537 (-) 15  13  14  12  16  18 
 WP2uvrA (-) 23  19  27  22  22  24 
 TA98 (+) 33  46  35  33  45  42 
 TA100 (+) 143 140  137  141  130  156 
 TA1535 (+) 20  23  21  17  23  20 
 TA1537 (+) 27  25  21  22  24  25 
 WP2uvrA (+) 38  33  39  35  38  35 

(-) = without metabolic activation; (+) = with metabolic activation

Conclusions:
Administration of MLCT oil did not result in any concentration-related increases in revertant colonies/plates in either strain of bacteria, with or without exogenous metabolic activation.
Executive summary:

The genetic toxicity of the test item, a mixture of medium and long-chain triacylglycerols (MLCT) of known composition, was investigated. The study was conducted in triplicate, following plate incorporation methods, with experiments performed with and without metabolic activation. The study was performed at concentrations of 0, 313, 625, 1250, 2500 and 5000 µg/plate. The studies without metabolic activation utilised AF-2 and NaN3 as positive control substances; the studies with metabolic activation utilised Benzo(a)pyrene as positive control substance. Solvent treatment groups were used as the negative control in all experiments. There were no increases in the frequency of revertant colonies recorded for any of the bacterial strains, with any dose of the test item, with or without metabolic activation (S9-mix). The vehicle control plates gave counts of revertant colonies within the normal range, and all positive control chemicals induced marked increases in the frequency of revertant colonies with and without metabolic activation. Therefore MLCT was considered to be non-mutagenic.

Data source

Reference
Reference Type:
publication
Title:
Safety evaluation of a medium- and long-chain triacylglycerol oil produced from medium-chain triacylglycerols and edible vegetable oil
Author:
Matulka RA, Noguchi O, Nosaka N
Year:
2006
Bibliographic source:
Food and Chemical Toxicology 44:1530-1538

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source of test material:
Nisshin Oillo Group, Ltd., Tokyo, Japan
- Composition: Caprylic acid (9.7%), capric acid (3.3%), palmitic acid (3.8%), stearic acid (1.7%), oleic acid (51.2%), linoleic acid (18.4%), linolenic acid (9.0%) and other fatty acids (2.9%)

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
Type and composition of metabolic activation system:
- concentration or volume of S9 mix and S9 in the final culture medium: 0.5 ml
Test concentrations with justification for top dose:
0, 313, 625, 1250, 2500 and 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: no data - vehicle control used
Controls
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
benzo(a)pyrene
other: NaN3, AF-2, ICR-191
Details on test system and experimental conditions:
NUMBER OF REPLICATIONS:
- Number of cultures per concentration: triplicate

METHOD OF TREATMENT/ EXPOSURE:
- Test substance added in agar (plate incorporation)

TREATMENT AND HARVEST SCHEDULE:
- Preincubation period, if applicable: 20 minutes
Rationale for test conditions:
A preliminary study was conducted.
Evaluation criteria:
Not stated.
Statistics:
Not stated.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
True negative controls validity:
not examined
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not determined
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
True negative controls validity:
not examined
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not determined
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
True negative controls validity:
not examined
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not determined
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
True negative controls validity:
not examined
Positive controls validity:
valid
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not determined
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
True negative controls validity:
not examined
Positive controls validity:
valid

Any other information on results incl. tables

  Dose (µg/plate)                 
 Strain 313  625  1250  2500  5000 
 TA98 (-) 24 25  17  17  22  24 
 TA100 (-) 129  140  123  136  122  136 
 TA1535 (-) 21  29  19  23  23  20 
 TA1537 (-) 15  13  14  12  16  18 
 WP2uvrA (-) 23  19  27  22  22  24 
 TA98 (+) 33  46  35  33  45  42 
 TA100 (+) 143 140  137  141  130  156 
 TA1535 (+) 20  23  21  17  23  20 
 TA1537 (+) 27  25  21  22  24  25 
 WP2uvrA (+) 38  33  39  35  38  35 

(-) = without metabolic activation; (+) = with metabolic activation

Applicant's summary and conclusion

Conclusions:
Administration of MLCT oil did not result in any concentration-related increases in revertant colonies/plates in either strain of bacteria, with or without exogenous metabolic activation.
Executive summary:

The genetic toxicity of the test item, a mixture of medium and long-chain triacylglycerols (MLCT) of known composition, was investigated. The study was conducted in triplicate, following plate incorporation methods, with experiments performed with and without metabolic activation. The study was performed at concentrations of 0, 313, 625, 1250, 2500 and 5000 µg/plate. The studies without metabolic activation utilised AF-2 and NaN3 as positive control substances; the studies with metabolic activation utilised Benzo(a)pyrene as positive control substance. Solvent treatment groups were used as the negative control in all experiments. There were no increases in the frequency of revertant colonies recorded for any of the bacterial strains, with any dose of the test item, with or without metabolic activation (S9-mix). The vehicle control plates gave counts of revertant colonies within the normal range, and all positive control chemicals induced marked increases in the frequency of revertant colonies with and without metabolic activation. Therefore MLCT was considered to be non-mutagenic.