Phenol, paraalkylation products with C10-15 branched olefins (C12 rich) derived from propene oligomerization, carbonates, calcium salts, overbased, sulfurized, including distillates (petroleum), hydrotreated, solvent-refined, solvent-dewaxed, or catalytic dewaxed, light or heavy paraffinic C15-C50

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

5th to 15th April 1985

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study following GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1985

Report date:

1985

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

not specified

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Phenol, paraalkylation products with C10-15 branched olefins (C12 rich) derived from propene oligomerization, carbonates, calcium salts, overbased, sulfurized including distillates (petroleum), hydrotreated, solvent-refined, solvent-dewaxed, or catalytic dewaxed, light or heavy paraffinic C15-C50

EC Number:

701-251-5

Molecular formula:

Formula for a representative structure is C36H58Ca2O4Sx where x = 1,2. Actual molecular formula is not possible to generate. Substance is a UVCB.

IUPAC Name:

Phenol, paraalkylation products with C10-15 branched olefins (C12 rich) derived from propene oligomerization, carbonates, calcium salts, overbased, sulfurized including distillates (petroleum), hydrotreated, solvent-refined, solvent-dewaxed, or catalytic dewaxed, light or heavy paraffinic C15-C50

Test material form:

liquid: viscous

Details on test material:

- Name and of test material (as cited in study report): Phenol, tetrapropenyl-, sulfurized, carbonates, calcium salts, overbased
- CAS number of test material (as cited in study report): 68784-26-9
- Physical state: Dark brown viscous liquid
- Other: The test substance was assumed to be 100% active ingredient. Stability and purity data were the responsibility of the sponsor.

Method

Species / strain

Species / strain / cell type:

other: S. typhimurium TA 98, TA 100 and TA 102 & E. coli WP2 uvrA

Details on mammalian cell type (if applicable):

Species and cell type: Male Sprague-Dawley rat, liver S-9 fractionQuantity: 25 µl/plate

Additional strain / cell type characteristics:

other: Histidine-deficient strains of Salmonella typhimurium and a tryptophan-deficient strain of Escherichia coli were grown.

Metabolic activation:

with and without

Metabolic activation system:

0.5 mL of 5% v/v S-9 mix per plate Induced or not induced: Aroclor 1254 induced, 500 mg/kg i.p.

Test concentrations with justification for top dose:

0, 0.033, 0.1, 0.33, 1.0, and 3.33 mg/plate, with and without S-9

Vehicle / solvent:

The vehicle used was 25% w/w Pluronic 127 (surfactant, CAS # 9003-11-6) in ethanol.

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: See table in materials and methods section below.

Details on test system and experimental conditions:

METHOD OF APPLICATION: All media and solutions were prepared as described by Maron and Ames (1983). Plate incorporation test.
Three replicate plates were evaluated for each treatment.

DURATION
The bacterial tester strain culture was grown by inoculating nutrient broth with a tester strain and incubating with shaking at ~37°C to a density of 1-2 x 10ˆ9 cells/ml. Without metabolic activation, 100 µl of tester strain and 100 µl of solvent or test material were added to 2.5 ml of molten selective top agar at ~45°C. With metabolic activation, 100 µl of tester strain, 100 µl of solvent or test material, and 0.5 ml of S-9 mix were added to 2.0 ml of molten selective top agar at ~45°C. After vortexing, the mixture was overlaid onto the surface of 25 ml of minimal bottom agar. After the overlay had solidified, the plates were inverted and incubated for 48 hours at ~37°C. The number of revertant colonies was counted and the presence and condition of the bacterial lawn was noted. Solvent and positive controls were run concurrently.

Test material was not completely miscible with the top agar at >5 mg/plate. The test material mixture for the highest dose level (10 mg/plate) was too viscous to deliver accurately.
Testing at the highest pipettable dose level (5 mg/plate) with 25-400 µl S-9/plate on strains TA98 and TA100 indicated that increasing the concentrations of S-9 did not signficantly increase the induction of revertants; therefore, 25 µl S-9/plate was used for the mutagenicity assay.

DETERMINATION OF CYTOTOXICITY
- Method: The test materials were checked for toxicity on strain TA100 with and without S-9 mix at dose levels of 0.003 to 10 mg/plate. No toxicity was observed.

Evaluation criteria:

The following criteria were met before the results of the mutagenicity assay were judged as positive or negative:
(1) the high dose showed some toxicity, or was 10 mg/plate or the limit of solubility;
(2) the spontaneous controls were within the laboratory's acceptable range;
(3) the positive controls produced at least a three-fold increase in the number of revertants over the values for the respective negative controls.

Results and discussion

Test results

Species / strain:

other: S. typhimurium TA 98, TA 100 and TA 102 & E. coli WP2 uvrA

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:

When tested at concentrations of 0.033 to 3.33 mg/plate the test material was not mutagenic to TA98, TA100, TA102, or E. Coli WP2 uvrA with or without metabolic activation provided by Aroclor-induced rat liver S-9. No cytotoxicity was observed at any concentration. The apparent increase in strain TA102 with S-9 metabolic activation is not considered biologically significant; the mean ± SEM for historical solvent control values is 325 ± 30.

The tester strains responded to the positive controls, known mutagens, as expected.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

First criterion for a valid test (as stated in the report) was not met. Dose of 5 mg/plate was the highest pipettable dose level and the solubility limit in top agar. Other test validity criteria were satisfied. No information was presented on condition of the background lawn, which would help assessment of cytotoxicity. Nevertheless, highest test dose of 3.33 mg/plate was close to the solubility limit and clearly not cytotoxic or mutagenic, based on revertants/plate data. The greatest mean number of revertants/plate over concurrent control among all test doses and all strains was 42% with S-9 and 19% without S-9. There was no indication of a dose-related decrease or increase of mean revertants/plate. Statistical results were not applicable.

Applicant's summary and conclusion

Conclusions:

Under the conditions tested, the test material was not mutagenic to TA98, TA100, TA102, or E. coli WP2 uvrA.

Executive summary:

In a GLP compliant study conducted in line with the standard methodology published by Ames, the test material was tested in the histidine-deficient strains of Salmonella typhimurium TA98, TA100, and TA102 and in the tryptophan-deficient strain of Escherichia coli WP2 uvrA at dose levels of 0.033 to 3.33 mg/plate with and without metabolic activation provided by Aroclor-induced rat liver S-9. The test material was suspended in 25% Pluronic 127 (w/w in ethanol). The suspension was miscible with the top agar. It was not cytotoxic at any concentration tested.

Under the conditions tested, the test material was not mutagenic to TA98, TA100, TA102, or E. coli WP2 uvrA.