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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: chromosome aberration
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1995
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Read across from structural analogue
Cross-referenceopen allclose all
Reason / purpose for cross-reference:
reference to same study
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
Justification for type of information:
Read across from structural analogue
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
reference to other study
Sex:
male
Genotoxicity:
negative
Toxicity:
no effects
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid

Table 7.6.2/2: In vivo micronucleus test results

Group

Concentration

MNF (%)

PCE (%)

 

CP (mg/kg bw)

PNE (mg/kg bw)

Distilled water

 

 

2.18 ± 0.58

52.90 ± 4.71

CP

30

-

36.68 ± 1.56*

40.74 ± 4.42*

PNE

-

2000

2.38 ± 1.18

53.95 ± 4.50

PNE + CP

30

500

27.31 ± 1.29*

51.13 ± 4.21*

PNE + CP

30

1000

18.40 ± 2.73*

47.58 ± 3.25***

PNE + CP

30

2000

8.88 ± 1.38*

42.50 ± 4.32***

PNE + CP

30

4000

10.53 ± 2.42*

40.40 ± 3.96***

 

CP: Cyclophosphamide; PNE: Pine needle extract

* p < 0.01, compared to distilled water control; * * p < 0.01, * * * p < 0.05, compared to CP groups.

There was a significant negative correlation (r = -0.9782, p < 0.05) between PNE dose and MNF in the PNE-treated groups in comparison to the CP positive group up to 2000 mg/kg bw of PNE. However, when the dose of PNE reached 4000 mg/kg bw, it exhibited some increase of MNF compared with the 2000 mg/kg bw group. This suggested that PNE inhibited the MNF in certain concentration ranges.

Conclusions:
Based on the read-across approach, the target substance did not induce micronuclei in mice.
Executive summary:

In an in vivo micronucleus test, ten male mice were injected intraperitoneally with Pine needle extract at the dose level of 2000 mg/kg bw and animals were sacrificed 30 h after the test item administration. Negative control (distilled water) and positive controls (cyclophosphamide at 30 mg/kg) were also included in the study. Antimutagenic effect of PNE was also studied with following dose levels at 10 males/group: (PNE + CP): 500 mg/kg bw + 30 mg/kg bw; 1000 mg/kg bw + 30 mg/kg bw; 2000 mg/kg bw + 30 mg/kg bw; 4000 mg/kg bw + 30 mg/kg bw.

Test material did not induce micronucleated PCE when compared with negative control. PNE could decrease the micronucleus frequency (MNF) compared with the cyclophosphamide (positive group) and the PNE + CP groups. Results showed that PNE could decrease the micronucleus induced by CP, and micronucleus frequency decreased markedly at the dose of 2000 mg/kg bw PNE (p < 0.01) demonstrating the antimutagenic effect. Positive control induced a statistically significant increase in micronucleated polychromatic erythrocytes indicating the validity of the study.

 

Based on the read-across approach, the target substance did not induce micronuclei in mice.

Reason / purpose for cross-reference:
reference to other study

Data source

Reference
Reference Type:
publication
Title:
Unnamed
Year:
1995

Materials and methods

Principles of method if other than guideline:
In an in vivo micronucleus test, mice injected intraperitoneally with Pine needle extract and animals sacrificed for examination of micronucleated polychromatic erythrocytes.
GLP compliance:
not specified
Type of assay:
micronucleus assay

Test material

Constituent 1
Reference substance name:
90082-72-7
Cas Number:
90082-72-7
IUPAC Name:
90082-72-7
Test material form:
not specified
Details on test material:
- Name of test material (as cited in study report): Pine needle extract (ointment); which is mainly composed of steroidal saponins, triterpenoidal saponins, terpenes and aromatic acids.
- Source: Jiangsu Institute of Cancer Research, China.

Test animals

Species:
mouse
Strain:
Balb/c
Sex:
male
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Animal Breeding Center of Nanjing University, Nanjing, China.
- Weight at study initiation: 18-22 g (at receipt)

Administration / exposure

Route of administration:
intraperitoneal
Duration of treatment / exposure:
30 h after test item administration
Frequency of treatment:
Single
Post exposure period:
None
Doses / concentrations
Remarks:
Doses / Concentrations:
2000 mg/kg bw
Basis:
nominal conc.
No. of animals per sex per dose:
10 males/dose
Control animals:
yes
Positive control(s):
- Positive control: Cyclophosphamide: 30 mg/kg bw
- Route of administration: Intraperitoneal

Examinations

Tissues and cell types examined:
Micronucleus frequency in polychromatic erythrocytes (PCE) was determined by the routine method of Schmid (1973). 1000 PCEs in each animal were examined for micronucleus.
Evaluation criteria:
No data
Statistics:
The data were statistically evaluated with the t-test.

Results and discussion

Test results
Sex:
male
Genotoxicity:
negative
Toxicity:
no effects
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid

Any other information on results incl. tables

Table 7.6.2/2: In vivo micronucleus test results

Group

Concentration

MNF (%)

PCE (%)

 

CP (mg/kg bw)

PNE (mg/kg bw)

Distilled water

 

 

2.18 ± 0.58

52.90 ± 4.71

CP

30

-

36.68 ± 1.56*

40.74 ± 4.42*

PNE

-

2000

2.38 ± 1.18

53.95 ± 4.50

PNE + CP

30

500

27.31 ± 1.29*

51.13 ± 4.21*

PNE + CP

30

1000

18.40 ± 2.73*

47.58 ± 3.25***

PNE + CP

30

2000

8.88 ± 1.38*

42.50 ± 4.32***

PNE + CP

30

4000

10.53 ± 2.42*

40.40 ± 3.96***

 

CP: Cyclophosphamide; PNE: Pine needle extract

* p < 0.01, compared to distilled water control; * * p < 0.01, * * * p < 0.05, compared to CP groups.

There was a significant negative correlation (r = -0.9782, p < 0.05) between PNE dose and MNF in the PNE-treated groups in comparison to the CP positive group up to 2000 mg/kg bw of PNE. However, when the dose of PNE reached 4000 mg/kg bw, it exhibited some increase of MNF compared with the 2000 mg/kg bw group. This suggested that PNE inhibited the MNF in certain concentration ranges.

Applicant's summary and conclusion

Conclusions:

Under the test conditions, Pine needle extract did not induce micronuclei in mice.
Executive summary:

In an in vivo micronucleus test, ten male mice were injected intraperitoneally with Pine needle extract at the dose level of 2000 mg/kg bw and animals were sacrificed 30 h after the test item administration. Negative control (distilled water) and positive controls (cyclophosphamide at 30 mg/kg) were also included in the study. Antimutagenic effect of PNE was also studied with following dose levels at 10 males/group: (PNE + CP): 500 mg/kg bw + 30 mg/kg bw; 1000 mg/kg bw + 30 mg/kg bw; 2000 mg/kg bw + 30 mg/kg bw; 4000 mg/kg bw + 30 mg/kg bw.

Test material did not induce micronucleated PCE when compared with negative control. PNE could decrease the micronucleus frequency (MNF) compared with the cyclophosphamide (positive group) and the PNE + CP groups. Results showed that PNE could decrease the micronucleus induced by CP, and micronucleus frequency decreased markedly at the dose of 2000 mg/kg bw PNE (p < 0.01) demonstrating the antimutagenic effect. Positive control induced a statistically significant increase in micronucleated polychromatic erythrocytes indicating the validity of the study.

 

Under the test conditions, Pine needle extract did not induce micronuclei in mice.