Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2017

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
Version / remarks:
Standards Established by the Minister of Labour in Accordance with the Industrial Safety and Health Law, Article 57-4-1”, (Notification No. 208, April 18, 2016 Ministry of Labour, Japan)
Guidelines for Toxicity Testing of New Chemical Articles”, (Notification 1221 No. 1 of Pharmaceutical and Food Safety Bureau, Ministry of Health, Labour and Welfare, 20151209 No. 1 of the Manufacturing Industries Bureau, Ministry of Economy, Trade and Industry, & No. 1512211 of Environmental Policy Bureau, Ministry of the Environment, Japan, on December 21, 2015)
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Reference substance name:
Reaction mass of N,N-Bis(2-hydroxypropyl)(C6-C18 even numbered and C18 unsatd)amides and bis(2-hydroxypropyl)ammonium myristate
Molecular formula:
No molecular formula. UVCB reaction mass.
IUPAC Name:
Reaction mass of N,N-Bis(2-hydroxypropyl)(C6-C18 even numbered and C18 unsatd)amides and bis(2-hydroxypropyl)ammonium myristate
Test material form:
liquid
Specific details on test material used for the study:
Supplier: MIC Trading Co., Ltd.
Amount Received: About 10 g
Date of Receipt: April 19, 2017
Name: Coconut diisopropanolamide
Another Name: COCAMIDE DIPA
CAS Number: 68855-69-6
Lot Number: 55780B17

Method

Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Vehicle / solvent:
Name: 1,4-dioxane
Manufacturer: Wako Pure Chemical Industries, Ltd.
Lot Number: DSH1946
Specification: JIS Reagent Special Grade, 99.5% or more
Controls
Untreated negative controls:
yes
Remarks:
1,4-dioxane, which was used for preparation of the test solution, was selected as the negative control article.
Positive controls:
yes
Positive control substance:
sodium azide
benzo(a)pyrene
other: 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide (AF-2); 2-Methoxy-6-chloro-9-[3-(2-chloroethyl)- aminopropylamino]acridine・2HCl (ICR-191); 2-Aminoanthracene (2AA
Details on test system and experimental conditions:
The test article (0.300 mL) was put into a sterilized test tube for preparation, and weighed. The weighed value was converted using the conversion factor of 0.86 calculated from the purity of 86%, the amount of vehicle in the test article suspension at the highest concentration of 100 mg/mL using sonication was calculated. Then, 1,4-dioxane was added to prepare the test article suspension at 100 mg/mL.
This test article suspension was diluted 6 times using a common ratio of 4 to prepare the test article suspension at a total of 7 concentrations: 50, 12.5, 3.13, 0.781, 0.195, 0.0488 and 0.0122 mg/mL.

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
Precipitation and Coloration by Test Article -
Neither precipitation nor coloration by the test article on the plate was observed at any dose levels irrespective of the presence/absence of metabolic activation

Growth Inhibition -
In the observation of bacterial background lawn using a stereoscopic microscope, growth inhibition was observed at 9.77 µg/plate or more for S. typhimurium TA1535 and TA1537 without metabolic activation, at 19.5 µg/plate or more for S. typhimurium TA98, TA100 and E. coli WP2 uvrA without metabolic activation, at 156 µg/plate or more for S. typhimurium TA100, TA1535 and TA1537 with metabolic activation and at 313 µg/plate or more for S. typhimurium TA98 and E. coli WP2 uvrA with metabolic activation

Number of Revertant Colonies
In the dose-finding test and two main tests, there was neither increase in the number of revertant colonies of two-fold or more in comparison with that of the negative control group nor dose-response for any strains irrespective of the presence/absence of metabolic activation

Applicant's summary and conclusion

Conclusions:
In the dose-finding test and the two main tests, there was neither increase in the number of revertant colonies of two-fold or more in comparison with that of the negative control group nor dose-response for any strains irrespective of the presence/absence of metabolic activation.
Since two-fold or more increase in the number of revertant colonies in comparison with the negative control group was observed in the positive control group for each tester strain, it was judged that the reactions of the bacterial strains to the mutagenic agents were suitable and thus the study was conducted appropriately.
In conclusion, COCAMIDE DIPA was judged to have no gene mutation inducibility (negative) under the conditions of this study.