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Toxicological information

Genetic toxicity: in vivo

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Administrative data

Endpoint:
in vivo mammalian cell study: DNA damage and/or repair
Remarks:
Type of genotoxicity: DNA damage and/or repair
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study is classified as reliable without restrictions because it was carried out in a method equivalent/similar to OECD TG 479 with modifications.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1988
Report Date:
1988

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
other: OECD 479
Deviations:
yes
Remarks:
Test was conducted in vivo and not in vitro as recommended by the guideline
GLP compliance:
yes
Type of assay:
sister chromatid exchange assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
IUCLID4 Test substance: other TS: Hydrodesulfurised kerosine, sample API 81-07, CAS No. 64742-81-0
Clear Liquid
Carbon Number Range: C9 through C16
Boiling Range: 150 degrees Celsius to 290 degrees Celsius
Storage: closed container in a cool, dark location.
Substance was a clear liquid.

Test animals

Species:
mouse
Strain:
B6C3F1
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Breeding Laboratories, Inc.
- Weight at study initiation: Toxicity Study: Male 21-28 grams. Female 12-22 grams. SCE Assay: Male 22-31 grams. Female 18-23 grams.
- Assigned to test groups randomly: No.
- Housing: Plastic autoclavable cages with wire lids. Hardwood chips for bedding.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: Quarantined for 10-14 days.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23-26 degrees Celsius
- Humidity (%): relative
- Photoperiod (hrs dark / hrs light): 12/ 12

Administration / exposure

Route of administration:
intraperitoneal
Vehicle:
- Vehicle(s)/solvent(s) used: corn oil.
Duration of treatment / exposure:
The animals were exposed for 20-22 hours
Frequency of treatment:
The animals were administered a single dose.
Doses / concentrations
Remarks:
Doses / Concentrations:
400, 2000 & 4000 mg/kg
Basis:
no data
No. of animals per sex per dose:
SCE Assay: five males and five females.
Positive control(s):
cyclophosphamide
- Preparation: dissolved in distilled water at concentration 1 mg/ml
- Route of administration: was administered intraperitoneally

API-81-15
Preparation: Dissolved in corn oil at concentration 400 mg/ml

Examinations

Tissues and cell types examined:
Sister Chromatid Exchanges per individual animals
Details of tissue and slide preparation:
Cells were collected by centrifugation, resuspended to opalescene in fresh fixative.
Two to four drops of fixed cells were dropped onto a wet slide and air dried. Slides were stained by Hoechst 33258-black light-Giemsa technique. Two to five slides were prepared from each animal.

Metaphase cells were examined under oil immersion without prior knowledge of treatment groups.
Evaluation criteria:
The number of SCEs per total second-division metaphase cells scored were recorded for each group, along with the range of SCEs/metaphase for animals within groups.
Statistics:
Kruskall Wallis test was used. Mann Whitney Test was used if required.
Test article is considered to induce a positive response if a dose-related increased in SCEs/metaphase is observed relative to the vehicle control (p< or = 0.05 Mann Whitney)

Results and discussion

Test resultsopen allclose all
Sex:
male
Genotoxicity:
positive
Sex:
female
Genotoxicity:
negative

Any other information on results incl. tables

The high dose male mice were lethargic shortly after dose administration and on the following day. In the high dose males and females and the mid-dose males there was a slight weight loss between the time pretreatment body weights were measured and when the animals were treated with colchicine on the following day.
The body weight changes are shown below:

Treatment

Sex

% change (+ or -)

Corn oil

 

24 hr

10 ml/kg

M

 0

 

F

- 2.9

API 81-07

M

- 1.4

400 mg/kg

F

- 0.5

2000 mg/kg

M

- 3.5

 

F

- 1.9

 

 

 

4000 mg/kg

M

- 4.3

 

F

- 4.7

API81-15

M

 0

4000 mg/kg

F

+ 3.8

Cyclophosphamide

 

 

10 mg/kg

M

+ 2.6

 

F

- 3.8


The SCEs were counted in 50 second-division metaphase cells and these data are summarized in the following table.


Treatment

Sex

Range of mean SCEs/cell for Individual Animals

Average SCEs/cell per mouse (a)

Corn oil

M

4.68 - 5.84

5.44±0.47 (5.64)

 

F

5.28 - 7.36

6.25±0.86 (6.06)

API81-07

 

 

 

400 mg/kg

M

6.76 - 10.46

8.26±1.51 (5.64)**

 

F

6.98 - 8.26

7.56±0.52 (7.56)

2000 mg/kg

M

5.68 - 7.58

6.74±0.87 (6.86)**

 

F

6.8 - 9.84

8.26±1.22 (8.54)

4000 mg/kg

M

5.72 - 7.66

6.86±0.75 (7.06)*

 

F

6.56 - 12.3

9.20±2.49 (9.88)

API81-15

 

 

 

4000 mg/kg

M

6.68 - 9.28

7.94±0.93 (7.94)**

 

F

7.28 - 8.54

7.86±0.58 (7.56)*

Cyclophosphamide

 

 

 

10 mg/kg

M

36.6 - 44.18

40.3±3.53 (38.5)**

 

F

18.34 - 31.64

25.5±5.44 (25.06)**

(a) Mean ± standard deviation (Median SCEs/cell)

*P < 0.05

** P < 0.01 (Mann Whitney test)


It was concluded that the negative and positive controls fulfilled the requirements and that API 81-07 was positive in male mice.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): positive positive in male mice, negative in female mice
No statistically significant effect was observed in the female mice, whereas the female mice responded positively to administration of both cyclophosphamide (10 mg/kg) and a carcinogenic heavy fuel oil (4000 mg/kg) which were used as positive control.
Under the conditions described in this report, the test article hydrodesulfurised kerosine does induce a significant increase in bone marrow SCEs in male B6C3F1 mice. However, the doses of the test article were rather high (400, 2000 and 4000 mg/kg) and the males in the highest dose groups showed signs of toxicity (lethargy and weight loss) on the day of the administration of the kerosine and the day after (when they were sacrificed).
Executive summary:

In a mammalian cell SCE assay, four hours after agar-coated BrdUrd pellet implantation the mice were given hydrodesulfurised kerosine as a single i.p dose at doses of either 400, 2000 or 4000 mg/kg in corn oil at a dose volume of 10 mL/kg (doses based a range finding study). A solvent control group was given corn oil at a dose of 10 mL/kg. Two positive control groups were used. One received cyclophosphamide i.p. at a dose of 10 mL/kg and the other received API 81-15 (a carcinogenic heavy fuel oil component) at a dose level of 4 g/kg, also at an injection rate of 10 mL/kg.

 

Positive control group API 81 -15 induced a significant increase in SCEs/cell/mouse relative to the vehicle control, as did the cyclophophamide. The results of the assay indicate that under the conditions described in this report, the test article hydrodesulfurised kerosine does induce a significant increase in bone marrow SCEs in male B6C3F1 mice, but not in female mice. The female mice responded positively to administration of both cyclophosphamide (10 mg/kg) and a carcinogenic heavy fuel oil (4000 mg/kg) which were used as positive control. The doses of the test article were rather high (400, 2000 and 4000 mg/kg) and the males in the highest dose groups showed signs of toxicity (lethargy and weight loss) on the day of the administration of the kerosine and the day after (when they were sacrificed).

 

This study received a Klimisch score of 1 and is classified as reliable without restriction because it was carried out in a method equivalent/similar to OECD TG 479 with modifications.