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Toxicological information

Genetic toxicity: in vivo

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Administrative data

Endpoint:
in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
The test was based on OECD Guideline for Testing of Chemicals: (No. 474 Mammalian Erythrocyte Micronucleus Test, July 1997). This final report and the original raw data were reviewed for adherence to ISO/IEC17025: 2005 General Requirements for the competence of Testing Calibration Laboratories. OECD Guideline for Testing of Chemicals: (No. 474 Mammalian Erythrocyte Micronucleus Test, July 1997); State Environmental Protection Administration: The Guidelines for the Testing of Chemicals ; Ministry of Health, 2005: Technical standards for testing of chemicals.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of assay:
transgenic rodent mutagenicity assay

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Species:
mouse
Strain:
other: NIH
Sex:
male/female
Details on test animals and environmental conditions:
Animals used: SPF NIH mice were provided by Guangdong Medical Laboratory Animal Center, certified animal No.: SCXK (guangdong) 2008-0002, 5 males and 5 females per group.

Environmental conditions
Test Room: In the Center
Animal house conditions: The test facility was an air-conditioned room with 12h artificial fluorescent light and 12h dark.
Temperature range: 20~25oC.
Humidity range: 40~70%.

Husbandry Practices
Caging: Stainless steel cages were used. Autoclaved clean dry corncob was used as the bedding material. Animals were housed in one group according to sex in cages.
Water bottle: Each cage was supplied with a polypropylene water bottle with a stainless steel nozzle.
Sanitation: Bedding material was changed daily.
Food and water: Standard pellet feed supplied by Guangdong Medical Laboratory Animal Center and ultra-pure filtered sterilized water were provided to the animals freely.
Frequency of providing feed and drinking water: Both drinking water and feed were provided ad libitum.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
material dissovled in distilled water.
Duration of treatment / exposure:
Date of testing: 30 June 2010 - 20 August 2010
108 days
Frequency of treatment:
doses administered at 210, 665, 2100 mg/kg at a volume of 10 ml/kg twice in 24 hours
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
210 mg/kg
Basis:
actual ingested
Remarks:
Doses / Concentrations:
665 mg/kg
Basis:
actual ingested
Remarks:
Doses / Concentrations:
2100 mg/kg
Basis:
actual ingested
No. of animals per sex per dose:
5 males and 5 females
Control animals:
yes
Positive control(s):
Animals of positive control group were given 40mg/kg cyclophosphamide dissolved in normal saline, by intraperitoneal injection and the volume was 10ml/kg. Samples were collected once between 18 and 24 hours following the final treatment for the bone marrow.
Negative control group were given distilled water instead of tested compounds.

Examinations

Tissues and cell types examined:
Bone Marrow Tissue

Results and discussion

Test results
Sex:
male/female
Genotoxicity:
negative
Toxicity:
no effects
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
RESULTS

Toxicity signs: In the dose levels of 210, 665 and 2100 mg/kg, none of the animals displayed abnormalitied during the observation period.

Micronucleated immature erythrocytes: Compared with negative control group, the micronucleus rates and the micronucleus cell frequencies of the test substance group showed no obvious increase (P<0.05). On the other hand, the micronucleus rates and the micronucleus cell frequencies of positive group were higher than negative groups (P<0.01). The detailed data showed in table 1-6.

Proportion of immature erythrocytes among total erythrocytes: The proportion of immature among total erythrocytes of each group had no difference.

Any other information on results incl. tables

Micronucleus rates and Proportion of immature among total erythrocytes

    Table 1

Group

(mg/kg)

Number of animal

Number of cells analysed

Number of micronucleated immature erythrocytes

Proportion of micronucleated immature erythrocytes ()

Number of micronucleated immature erythrocytes

(x±s)

Proportion of immature among total erythrocytes

P

Negative control group

10

20000

3

0.3

0.3±0.67

 

210

10

20000

8

0.8

0.8±0.82

0.353

665

10

20000

10

1.0

1.0±1.15

0.684

2100

10

20000

10

1.0

1.0±1.25

0.143

Positive group

10

20000

352

17.6**

35.2±19.50

0.353

           ** Compared with negative control group,P<0.01

 

 

 

 

 

 

 

The detailed data of each animal ( negative control group )

Table 2  

Gender

No. of animal

Number of cells analysed

Number of micronucleated immature erythrocytes

Proportion of micronucleated immature erythrocytes ()

Proportion of immature erythrocytes among total erythrocytes (%)

4

2000

0

0

52.0

17

2000

0

0

67.0

18

2000

2

1

56.0

23

2000

0

0

54.8

28

2000

0

0

70.0

1

2000

1

0.5

64.0

19

2000

0

0

69.0

20

2000

0

0

50.0

26

2000

0

0

76.0

28

2000

0

0

64.0

 

 

 

 

 

 

The detailed data of each animal (210mg/kg )

Table 3

Gender

No. of animal(动物号)

Number of cells analysed

Number of micronucleated immature erythrocytes

Proportion of micronucleated immature erythrocytes ()

Proportion of immature erythrocytes among total erythrocytes(%)

8

2000

0

0

81.0

14

2000

0

0

57.0

24

2000

0

0

54.0

25

2000

2

1

69.0

26

2000

0

0

51.5

7

2000

2

1

70.0

8

2000

2

1

60.5

18

2000

0

0

71.0

23

2000

1

0.5

71.9

24

2000

1

0.5

69.5

 

 

 

 

 

The detailed data of each animal (665mg/kg )

Table 4

Gender

No. of animal

Number of cells analysed

Number of micronucleated immature erythrocytes

Proportion of micronucleated immature erythrocytes ()

Proportion of immature erythrocytes among total erythrocytes(%)

6

2000

0

0

51.0

16

2000

0

0

61.5

12

2000

0

0

66.5

13

2000

3

1.5

60.0

57

2000

0

0

68.0

3

2000

1

0.5

53.0

4

2000

2

1

71.0

12

2000

2

1

84.0

15

2000

0

0

70.0

16

2000

2

1

59.5

 

 

 

 

 

The detailed data of each animal (2100mg/kg )

Table 5

Gender

No. of animal

Number of cells analysed

Number of micronucleated immature erythrocytes

Proportion of micronucleated immature erythrocytes ()

Proportion of immature erythrocytes among total erythrocytes (%)

1

2000

0

0

70.0

9

2000

3

1.5

75.6

10

2000

0

0

73.0

19

2000

3

1.5

75.6

20

2000

2

1

62.0

5

2000

0

0

49.5

6

2000

1

0.5

73.0

10

2000

1

0.5

81.0

13

2000

0

0

62.5

17

2000

0

0

67.5

 

 

 

 

 

The detailed data of each animal (positive group )

Table 6

Gender

No. of animal

Number of cells analysed

Number of micronucleated immature erythrocytes

Proportion of micronucleated immature erythrocytes ()

Proportion of immature erythrocytes among total erythrocytes(%)

3

2000

56

28

58.0

5

2000

65

32.5

62.0

7

2000

44

22

63.5

15

2000

29

14.5

60.3

27

2000

27

13.5

60.0

2

2000

8

4

45.0

9

2000

11

5.5

50.5

14

2000

50

25

79.5

25

2000

18

9

32.5

27

2000

44

22

65.0

 

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): negative
Pepton 22 has been show to be not mutagenic