tris(2-octyldodecyl) citrate

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:

28 April 1993 to 5 July 1993

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1993

Report date:

1993

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 (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

1,2,3-tris(2-octyldodecyl) 2-hydroxypropane-1,2,3-tricarboxylate

EC Number:

603-121-2

Cas Number:

126121-35-5

Molecular formula:

C66H128O7

IUPAC Name:

1,2,3-tris(2-octyldodecyl) 2-hydroxypropane-1,2,3-tricarboxylate

Test material form:

liquid

Details on test material:

Tris(octyldodecyl) citrate 99.87 %
Bis(octyldodecyl) citrate 0.13 %

Specific details on test material used for the study:

SAMPLE NUMBER: #934536 Citmol 320, EZ 93-004, Lot#21519 Five Ames strains both with and without metabolic activation (S9-rat liver microsomes).
Sample Characteristics: Received in a glass bottle as a clear, colorless, viscous liquid.
Solvent: Methylethylketone (MEK) + 10% Tween 80.
Concentration Range: 0, 100, 500, 1000, 500.J and 10,000 ) ug/plate.
Sample Plating: 0.1 ml was plated in triplicate.
Phase: Clear solution.
Plate Counting: After incubation for 48 hours, plates were counted with a Biotran III.

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

mammalian liver post-mitochondrial fraction (S-9)

Test concentrations with justification for top dose:

The sponsor may request that the dose range should be determined by first carrying out a cytotoxicity study with strain TAlOO. Depending upon solubility characteristics, the test article will be evaluated for direct toxicity at doses of 0, 5, 10, 50, 100, 500, 1000, 5000 and 10000 J..Lg/plate. Each dose of test article will be evaluated relative to the solvent control in order to determine the concentration which causes a 50% reduction in the spontaneous background level of TAlOO revertants.  The test doses will be plated  in duplicate both with and without activation against strain TAlOO (density of 108-109/ml) on Vogel-Bonner minimal media plates following the usual plating procedure for the Ames Test.
Liquids will be tested as received (neat) and then at appropriate dilutions ( 112 logs) to determine the non-toxic dose range. The Ames Test will then be carried out over dose levels in the non-toxic range.
Following the 48-hour incubation, .the numb_, er of revertant colonies per plate will be counted with a Biotran III automated colony counter. Toxicity is suggested by the absence of a confluent bacterial lawn, the presence of pin point colonies, and/or a substantial decrease or lack of revertant colonies.
The concentration of test article giving 50% survival (based on revertant colony formation) will be used as the highest dose in the actual Ames Test in which a minimum of five dose levels of test article will be evaluated.  If no toxicity is observed in the prescreen, a maximum of 10000 ,ug/plate will be used as the highest dose, solubility permitting. Otherwise, the sample will be plated at the solubility limit in the specified solvent.

Vehicle / solvent:

Methylethylketone (MEK) + 10% Tween 80.

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

benzo(a)pyrene

mitomycin C

other: 2-aminoanthracene

Results and discussion

Test results

Test results 1

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

Positive controls validity:

valid

Test results 2

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

Positive controls validity:

valid

Test results 3

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

Positive controls validity:

valid

Test results 4

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

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

The test substance was not mutagenic against any of the tester strains either directly or following Inetabolic activation with S9 (+M).

Executive summary:

In this OECD 471 Ames test, the test substance was not found to be mutagenic against any of the tester strains either directly or following Inetabolic activation with S9 (+M).