Petroleum heavy fuel oil fraction, co-processed with renewable hydrocarbons of plant or animal origin

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study, GLP status not known. Adequate for assessment.

Justification for type of information:

Read across justification included in Section 13

Cross-reference

Reason / purpose for cross-reference:

read-across: supporting information

Reference

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study, GLP status not known. Adequate for assessment.

Justification for type of information:

Read across justification included in Section 13

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.12 (Mutagenicity - In Vivo Mammalian Erythrocyte Micronucleus Test)

Principles of method if other than guideline:

Male and female CD-1 mice received catalytically clarified oil as two consecutive daily doses, either by gavage or i.p. injection. Animals were sacrificed 24 hr after the second treatment. Bone marrow cells were then collected and mature red blood cells (normochromatic erythrocytes, NCE) and immature red cells (polychromatic erythrocytes, PCE) evaluated for cytotoxicity and micronucleus formation.

GLP compliance:

not specified

Type of assay:

micronucleus assay

Species:

mouse

Strain:

CD-1

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Canada (Quebec, Canada)
- Age at study initiation: 6-9 wk
- Weight at study initiation: 17-35 g

Route of administration:

oral: gavage

Vehicle:

Vehicle: corn oil

Details on exposure:

Dose selection was based on a range-finding (toxicity) study (0, 1, 2 or 3 g/Kg bw)

Duration of treatment / exposure:

2 consecutive daily doses, with sacrifice 24 hr after second dose

Frequency of treatment:

daily for 2 days

Post exposure period:

24 hr

Remarks:

Doses / Concentrations:
0, 188, 375, 750 or 1500 mg/Kg bw/d
Basis:
nominal conc.

No. of animals per sex per dose:

5 male, 5 female

Control animals:

yes, concurrent vehicle

Positive control(s):

Cyclophosphamide, 40 mg/Kg bw (gavage in water)

Tissues and cell types examined:

Bone marrow smears (femur) were prepared from five animals per sex per dose group, stained with acridine orange and examined under fluorescence microscopy. One thousand erythrocytes were evaluated per animal and the total number PCEs and NCEs recorded. One thousand PCEs were evaluated for the presence of micronuclei.

Statistics:

Data were analysed using ANOVA followed by Duncan's Multiple Range Test. Regression analysis was used to characterise any dose-response relationship present.

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

valid

Positive controls validity:

valid

There was no significant increase in the percentages of micronucleated PCE 24 hr after 2 consecutive daily doses of catalytically cracked clarified oil at doses of 188, 375, 750 or 1500 mg/Kg bw/d:

Control = 1.3 / 1000 PCE

188 mg/Kg bw = 1.8 / 1000 PCE

375 mg/Kg bw = 1.8 / 1000 PCE

750 mg/Kg bw = 3.0 / 1000 PCE

1500 mg/Kg bw = 2.3 / 1000 PCE

The mean percentage of PCEs for treated animals was 49-54% versus 50-55% for the controls i.e. there was no evidence of cytotoxicity and it is not known if the test substance reached the target tissue (guideline criterion).

A satisfactory response was obtained with the positive control substance (cyclophosphamide).

Conclusions:

Interpretation of results (migrated information): negative
Catalytically cracked clarified oil did not induce micronucleated polychromatic erythrocytes in mouse femoral bone marrow following two consecutivedaily oral doses of 188-1500 mg/Kg bw/d.

Executive summary:

The clastogenic potential of catalytically cracked clarified oil was evaluated in male and female mice after 2 consecutive daily oral treatments of 0, 188, 375, 750 ot 1500 mg/Kg bw/d. Femoral bone marrow was collected 24 hr following the second treatment, and polychromatic erythrocytes present evaluated for micronucleus formation.

Catalytically cracked clarified oil was not cytotoxic (as judged from no change in the PCE:NCE ratio) nor did it induce any statistically significant increase in the formation of micronucleated PCEs. An absence of cytotoxicity may indicate that the test substance did not reach the target tissue.

Data source

Reference

Reference Type:

publication

Title:

Assessment of the utility of the micronucleus test for petroleum-derived materials

Author:

Przygoda, R.T., McKee, R.H., Amoruso, M.A. and Freeman, J.J.

Year:

1999

Bibliographic source:

Mutation Research 438, 145-153

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.12 (Mutagenicity - In Vivo Mammalian Erythrocyte Micronucleus Test)

Principles of method if other than guideline:

Male and female CD-1 mice received catalytically clarified oil as two consecutive daily doses, either by gavage or i.p. injection. Animals were sacrificed 24 hr after the second treatment. Bone marrow cells were then collected and mature red blood cells (normochromatic erythrocytes, NCE) and immature red cells (polychromatic erythrocytes, PCE) evaluated for cytotoxicity and micronucleus formation.

GLP compliance:

not specified

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

64741-62-4

IUPAC Name:

64741-62-4

Test material form:

other: Viscous hydrocarbon liquid

Details on test material:

- Name of test material (as cited in study report): catalytically cracked clarified oil

Test animals

Species:

mouse

Strain:

CD-1

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Canada (Quebec, Canada)
- Age at study initiation: 6-9 wk
- Weight at study initiation: 17-35 g

Administration / exposure

Route of administration:

intraperitoneal

Vehicle:

Vehicle: corn oil

Duration of treatment / exposure:

2 consecutive daily doses, with sacrifice 24 hr after second dose

Frequency of treatment:

daily for 2 days

Post exposure period:

24 hr

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
0, 188, 375 or 750 mg/Kg bw/d
Basis:
nominal conc.

Doses / concentrations 2

Remarks:

Doses / Concentrations:
0, 750, 1500 or 3000 mg/Kg bw/d
Basis:
nominal conc.

No. of animals per sex per dose:

Study 1 (188-750 mg/Kg bw/d): 5 male, 5 female per dose level
Study 2 (750-3000 mg/Kg bw/d): 2 male, 2 female per dose level

Control animals:

yes, concurrent vehicle

Positive control(s):

Cyclophosphamide, 40 mg/Kg bw (gavage in water)

Examinations

Tissues and cell types examined:

Bone marrow smears (femur) were prepared from five animals per sex per dose group, stained with acridine orange and examined under fluorescence microscopy. One thousand erythrocytes were evaluated per animal and the total number PCEs and NCEs recorded. One thousand PCEs were evaluated for the presence of micronuclei.

Statistics:

Data were analysed using ANOVA followed by Duncan's Multiple Range Test. Regression analysis was used to characterise any dose-response relationship present.

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Remarks:

in both studies

Toxicity:

no effects

Vehicle controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

There was no significant increase in the percentages of micronucleated PCE 24 hr after 2 consecutive daily doses of catalytically cracked clarified oil at doses of 188 -750 or 750 -3000 mg/Kg bw/d. Results for study 2 were as follows:

Control = 1.25 / 1000 PCE

750 mg/Kg bw = 1.75 / 1000 PCE

1500 mg/Kg bw = 2.50 / 1000 PCE

3000 mg/Kg bw = 1.00 / 1000 PCE

 

The mean percentage of PCEs was decreased for treated animals from study 2 as follows:

Control = 48.8%

750 mg/Kg bw/d = 41.3%

1500 mg/Kg bw/d = 36.8%

3000 mg/Kg bw/d = 32.4%

(historical control range = 45 -59%)

The decrease in mean percentage PCE indicates that the test substance reached the target tissue (guideline criterion).

 

A satisfactory response was obtained with the positive control substance (cyclophosphamide).

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negative
Catalytically cracked clarified oil did not induce micronucleated polychromatic erythrocytes in mouse femoral bone marrow following two consecutive daily i.p. treatments of up to 3000 mg/Kg bw/d.

Executive summary:

The clastogenic potential of catalytically cracked clarified oil was evaluated in male and female mice following 2 consecutive daily treatments of up to 3000 mg/Kg bw/d. Femoral bone marrow was collected 24 hr following the second treatment, and polychromatic erythrocytes present evaluated for micronucleus formation. Mild cytotoxicity (as indicated by a decrease in the percentage of PCE present) was apparent following treatments of 1500 and 3000 mg/Kg bw/d but there was no increased formation of micronucleated PCEs.

The results indicate the catalytically cracked clarified oil is not clastogenic in mouse bone marrow following i.p. administration at doses up to 3000 mg/Kg bw/d.