Middle distillate from refinery proces cofeeding vegetable oil

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:

1988 to 1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: This study is classified as reliable with restrictions because it is an acceptable, well-documented study report.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1991

Report date:

1991

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

Modified Ames assay

Principles of method if other than guideline:

Modifications of the standard Ames assay include:
1. DMSO extractions of the test material dissolved in cyclohexane were used as opposed to the neat or solvent-diluted sample.
2. An eight-fold higher concentration of liver S-9 fraction from hamster rather than rat was used.
3. The NADP cofactor was increased from 4 to 8 mm.

GLP compliance:

not specified

Type of assay:

other: bacterial reverse mutation assay (Modified Ames test)

Test material

Test material information

Constituent 1

Reference substance name:

Blends of straight run gas oil and fluid catalytically cracked gas oil, diesel fuel (CAS# 68334-30-5)

IUPAC Name:

Blends of straight run gas oil and fluid catalytically cracked gas oil, diesel fuel (CAS# 68334-30-5)

Details on test material:

Name of test material (as cited in study report): Blend 55% straight run gas oil, 45% fluid catalytically cracked gas oil (MD 57389), Diesel fuel (MD57393), Blend 54% straight run gas oil, 46% cracked gas oil components (MD 57391), Blend 50% straight run gas oil, 50% cracked gas oil components (MD 57386), Straight run gas oil with 59% cracked gas oil (MD 57401), Straight run gas oil cracked gas oil blend (MD 57388)
- Substance type: petroleum-derived middle distillate
- Physical state: liquid
- Analytical purity: not provided
- Impurities (identity and concentrations): not provided
- Composition of test material, percentage of components: not provided
- Isomers composition: not provided
- Purity test date: not provided
- Lot/batch No.: sample code- MD 57389
- Expiration date of the lot/batch: not provided
- Stability under test conditions: not provided
- Storage condition of test material: stored indoors in stainless steel containers at 18°C

- Name of test material (as cited in study report): Diesel fuel (MD 57393)
- Substance type: petroleum-derived middle distillate
- Physical state: liquid
- Analytical purity: not provided
- Impurities (identity and concentrations): not provided
- Composition of test material, percentage of components: not provided
- Isomers composition: not provided
- Purity test date: not provided
- Lot/batch No.: sample code- MD 57393
- Expiration date of the lot/batch: not provided
- Stability under test conditions: not provided
- Storage condition of test material: stored indoors in stainless steel containers at 18°C

- Name of test material (as cited in study report): Blend 54% straight run gas oil, 46% cracked gas oil components (MD 57391)
- Substance type: petroleum-derived middle distillate
- Physical state: liquid
- Analytical purity: not provided
- Impurities (identity and concentrations): not provided
- Composition of test material, percentage of components: not provided
- Isomers composition: not provided
- Purity test date: not provided
- Lot/batch No.: sample code- MD 57391
- Expiration date of the lot/batch: not provided
- Stability under test conditions: not provided
- Storage condition of test material: stored indoors in stainless steel containers at 18°C

- Name of test material (as cited in study report): Blend 50% straight run gas oil, 50% cracked gas oil components (MD 57386)
- Substance type: petroleum-derived middle distillate
- Physical state: liquid
- Analytical purity: not provided
- Impurities (identity and concentrations): not provided
- Composition of test material, percentage of components: not provided
- Isomers composition: not provided
- Purity test date: not provided
- Lot/batch No.: sample code- 57392
- Expiration date of the lot/batch: not provided
- Stability under test conditions: not provided
- Storage condition of test material: stored indoors in stainless steel containers at 18°C

- Name of test material (as cited in study report): Straight run gas oil with 59% cracked gas oil (MD 57401)
- Substance type: petroleum-derived middle distillate
- Physical state: liquid
- Analytical purity: not provided
- Impurities (identity and concentrations): not provided
- Composition of test material, percentage of components: not provided
- Isomers composition: not provided
- Purity test date: not provided
- Lot/batch No.: sample code- MD 57401
- Expiration date of the lot/batch: not provided
- Stability under test conditions: not provided
- Storage condition of test material: stored indoors in stainless steel containers at 18°C

- Name of test material (as cited in study report): Straight run gas oil cracked gas oil blend (MD 57388)
- Substance type: petroleum-derived middle distillate
- Physical state: liquid
- Analytical purity: not provided
- Impurities (identity and concentrations): not provided
- Composition of test material, percentage of components: not provided
- Isomers composition: not provided
- Purity test date: not provided
- Lot/batch No.: sample code- MD 57388
- Expiration date of the lot/batch: not provided
- Stability under test conditions: not provided
- Storage condition of test material: stored indoors in stainless steel containers at 18°C

Method

Target gene:

gene for histadine synthesis

Species / strain

Species / strain / cell type:

S. typhimurium TA 98

Details on mammalian cell type (if applicable):

- Type and identity of media: Oxoid 32
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: not reported
- Periodically checked for karyotype stability: not reported
- Periodically "cleansed" against high spontaneous background: not reported

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S-9 mix prepared with the livers of Syrian-Golden hamsters

Test concentrations with justification for top dose:

0, 1, 3, 5, 7, 10, 15, 20, 25, 40, 40, 50, 60 µl/plate

Vehicle / solvent:

cyclohaxane and dimethylsulfoxide (DMSO)

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: benzo(a)pyrene 2-aminoanthracene and 2-nitrofluorene

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: not reported
- Exposure duration: 48 hours
- Expression time (cells in growth medium): 48 hours
- Selection time (if incubation with a selection agent): not reported
- Fixation time (start of exposure up to fixation or harvest of cells): 48 hours


NUMBER OF REPLICATIONS: not reported


NUMBER OF CELLS EVALUATED: not reported

Evaluation criteria:

Spontaneous and solvent control reversion rates for TA98 should fall between 30 and 55 revertants/plate for the assay to be considered acceptable. The single dose of Stock 642-100'' CNN (standard reference material) at 50 µl must fall between 100-200 revertants/plate.

Statistics:

The mean number of revertants/plate for each dose was calculated. The Student-Newman Keuls Test was preformed if a dose-related doubling relative to the mean solvent control was not reached. Nonlinear regression, using the SAS NLIN procedure of Myers et al. was used to determine the slope of the dose response curve, if a doubling was reached.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

other: marginally mutagenic; MI- 1.7

Cytotoxicity / choice of top concentrations:

not specified

Test results 2

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

positive

Remarks:

MI-2.0

Cytotoxicity / choice of top concentrations:

not specified

Test results 3

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

positive

Remarks:

MI- 2.3

Cytotoxicity / choice of top concentrations:

not specified

Test results 4

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

positive

Remarks:

MI-3.9

Cytotoxicity / choice of top concentrations:

not specified

Test results 5

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

positive

Remarks:

MI-7.6

Test results 6

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

positive

Remarks:

Mutagenicity index (MI)- 9.0

Cytotoxicity / choice of top concentrations:

not specified

Remarks on result:

other: other: Diesel fuel (MD 57393)

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

A dose-related increase in revertants compared to the mean solvent control value occurred in the blend 55% straight run gas oil, 45% fluid catalytically cracked gas oil substance (MD 57389); the blend 54% straight run gas oil, 46% cracked gas oil components substance (MD 57391); the Blend 50% straight run gas oil, 50% cracked gas oil components substance (MD 57386); the straight run gas oil with 59% cracked gas oil (MD 57401); and the straight run gas oil cracked gas oil blend (MD 57388). The mutagenicity index for these substances ranged from 2.0 to 9.0. The diesel fuel (MD 57393) was marginally mutagenic with an MI of 1.7.

Table 1. Mutagenicity Indices of Test Materials
Test Material Type	Sample Number	Mutagenicity Index
Blend 55% straight run gas oil, 45% fluid catalytically cracked gas oil	MD 57389	9.0
Diesel fuel	MD 57393	1.7
Blend 54% straight run gas oil, 46% cracked gas oil components	MD 57391	2.3
Blend 50% straight run gas oil, 50% cracked gas oil components	MD 57386	7.6
Straight run gas oil with 59% cracked gas oil	MD 57401	3.9
Straight run gas oil cracked gas oil blend	MD 57388	2.0

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
positive based on the amount of cracked material present in the blend

A dose-related increase in revertants compared to the mean solvent control value occurred in the blend 55% straight run gas oil, 45% fluid catalytically cracked gas oil substance (MD 57389); the blend 54% straight run gas oil, 46% cracked gas oil components substance (MD 57391); the blend 50% straight run gas oil, 50% cracked gas oil components substance (MD 57386); the straight run gas oil with 59% cracked gas oil (MD 57401); and the straight run gas oil cracked gas oil blend (MD 57388). The mutagenicity index for these substances ranged from 2.0 to 9.0. These substances are considered genotoxic materials. The diesel fuel (MD 57393) was marginally mutagenic with an MI of 1.7.

Executive summary:

In a genetic toxicity study, a modified Ames test was conducted using six different types of petroleum-derived middle distillate test substances [blend 55% straight run gas oil, 45% fluid catalytically cracked gas oil substance (MD 57389); blend 54% straight run gas oil, 46% cracked gas oil components substance (MD 57391); blend 50% straight run gas oil, 50% cracked gas oil components substance (MD 57386); straight run gas oil with 59% cracked gas oil (MD 57401); straight run gas oil cracked gas oil blend (MD 57388); and diesel fuel (MD 57393)]. DMSO test article extracts were applied mixed with phosphate buffer (non-activating) or S-9 mix (activating) andSalmonella typhimuriumbroth culture, using the preincubation method. The pre-incubated mixture was overlayed on toVogel-Bonner Minimal E medium plus 1.5% bacteriological grade agar and 2% glucose in Nunc Petri plates. The plates were allowed to incubate for 48 hours. After the incubation period, colonies of histidine protrophs in test and control plates were enumerated on a Artek automatic colony counter. The condition of the background lawn of bacterial growth was confirmed by visual observation with a Zeiss stereozoom microscope. The same procedures were used for the positive controls (2 -aminoanthracene, benzo (a) pyrene, and 2 -nitrofluorene).

A dose-related increase in revertants compared to the mean solvent control value occurred in the blend 55% straight run gas oil, 45% fluid catalytically cracked gas oil substance (MD 57389); the blend 54% straight run gas oil, 46% cracked gas oil components substance (MD 57391); the blend 50% straight run gas oil, 50% cracked gas oil components substance (MD 57386); the straight run gas oil with 59% cracked gas oil (MD 57401); and the straight run gas oil cracked gas oil blend (MD 57388). The mutagenicity index for these substances ranged from 2.0 to 9.0. These substances are considered genotoxic materials. The diesel fuel (MD 57393) was marginally mutagenic with an MI of 1.7.

This study received a Klimisch score of 2 and is classified as reliable with restrictions because it is an acceptable and a well-documented study report.