citric acid, esters with dodecan-1-ol

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:

19.06.2016-20.02.2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

21 July 1997

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Didodecyl hydrogen citrate

EC Number:

247-146-3

EC Name:

Didodecyl hydrogen citrate

Cas Number:

25637-88-1

Molecular formula:

C30H56O7

IUPAC Name:

didodecyl hydrogen citrate

Constituent 2

Reference substance name:

Tridodecyl 2-hydroxypropane-1,2,3-tricarboxylate

EC Number:

265-671-6

EC Name:

Tridodecyl 2-hydroxypropane-1,2,3-tricarboxylate

Cas Number:

65277-53-4

Molecular formula:

C42H80O7

IUPAC Name:

tridodecyl citrate

Constituent 3

Reference substance name:

Dodecyl dihydrogen 2-hydroxypropane-1,2,3-tricarboxylate

EC Number:

278-950-2

EC Name:

Dodecyl dihydrogen 2-hydroxypropane-1,2,3-tricarboxylate

Cas Number:

78568-45-3

Molecular formula:

C18H32O7

IUPAC Name:

2-[2-(dodecyloxy)-2-oxoethyl]-2-hydroxysuccinic acid

Test material form:

solid

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

Metabolic activation system:

Mammalian liver post mitochondrial fraction (S9)

Test concentrations with justification for top dose:

First experiment: 62, 185, 556. 1667, 5000 µg/plate
Second experiment: rejected because of technical problems during performance
Third experiment: 3.7, 11.1, 33.3, 100, 300 µg/plate

Vehicle / solvent:

- solvent used: acetone
- Justification for choice of solvent: maximum concentration of the substance could be achieved

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

no

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

9-aminoacridine

2-nitrofluorene

sodium azide

other: 2-aminoanthracene (2-AA), benzo[a]pyrene (Be[a]P)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)


DURATION
- Preincubation period: 48 hours at 37 °C in the dark

NUMBER OF REPLICATIONS: 3 per concentration per strain

Evaluation criteria:

Biologically relevant response: If the number of revertants is at least twice the spontaneous reversion rate in one of the strains and/or if there is a concentration related increasing number of revertants over the range tested

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

not determined

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

not determined

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 5000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

not determined

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 5000 µg/plate

Test results 4

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

not determined

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 5000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

not determined

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 5000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Table 1: 1^stexperiment

Ratio mean revertant numbersconcentration/control
Concen-tration [µg/plate]	TA 1535	TA 1537	TA 98	TA 100	WP2 uvrA	TA 1535	TA 1537	TA 98	TA 100	WP2 uvrA
	without S9					with S9
62	1.2	1.1	1.2	0.9	1.0	1.1	0.8	0.6	1.2	1.3
185	1.0	1.2	0.8	0.6	0.4	0.7	0.8	0.8	0.9	0.6
556 P	0.7	1.8	0.9	0.8	0.8	0.6	1.0	0.8	1.1	0.8
1667 P	0.8	1.2	0.4	0.5	1.1	0.6	0.8	0.6	1.1	0.8
5000 P	0.7	0.3	0.7	0.2	0.6	0.7	0.6	0.3	0.8	0.8

 

P: precipitation

 

Table 3: 3^rdexperiment

Ratio mean revertant numbersconcentration/control
Concen-tration [µg/plate]	TA 1535	TA 1537	TA 98	TA 100	WP2 uvrA	TA 1535	TA 1537	TA 98	TA 100	WP2 uvrA
	without S9					with S9
3.7	1.0	1.4	1.0	1.0	1.1	1.0	0.5	0.8	1.3	1.0
11.1	0.9	0.8	0.4	1.0	1.1	0.8	1.0	1.1	0.9	1.0
33.3	0.9	1.0	0.9	0.9	0.9	0.8	1.3	1.0	1.0	0.9
100	0.9	1.3	0.7	0.9	0.9	0.9	0.9	0.9	0.9	0.9
300	0.7	1.1	0.8	0.9	1.0	0.8	0.6	0.6	0.6	1.0

 

Results from experiment 2 were rejected because of technical problems during performing.

Applicant's summary and conclusion

Conclusions:

No gene mutations by base pair substitution or frame shifts in the genome of the used strains were observed. Therefore the test substance is considered to be non mutagenic under the conditions employed. The substance was also not toxic when tested at least up to the concentration of 300 µg/plate.

Executive summary:

The mutagenic potential of the substance was investigated in a bacterial reverse mutation assay according to OECD Guideline 471. Two experiments were performed in th presence and in the absence of a metabolic activation system (S9). Concentrations used were up to 5000 µg/plate. No gene mutations by base pair substitution or frame shifts in the genome of the used strains were observed. Therefore the test substance is considered to be non mutagenic under the conditions employed. The substance was also not toxic when tested at least up to the concentration of 300 µg/plate.