Indigofera tinctoria, ext.

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Type of information:

other: SCCS opinion /1439/11

Adequacy of study:

supporting study

Study period:

21 March 2001 – 23 January 2003

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Data source

Reference

Reference Type:

publication

Title:

SCCS Opinion on Indigofera tinctoria COLIPA no C170

Author:

Scientific Committee on Consumer Safety

Year:

2012

Bibliographic source:

978-92-79-30776-8

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Version / remarks:

1997

Deviations:

not specified

GLP compliance:

yes

Type of assay:

mammalian erythrocyte micronucleus test

Test material

Test material information

Test material form:

solid

Specific details on test material used for the study:

19556 Indigoblätter was investigated for the induction of micronuclei in bone marrow cells
of mice. The test item was prepared by pouring distilled water (70°C) over the test item (1
part test item:5 parts aqua distellata) and by stirring them with a glass stick until it got a
homogeneous paste. After 30 min without heating the paste was filtered using a cellulose
filter. This filtrate was used in the experiments in a concentration relating to 200 mg/ml of
the original test item. 10 mg/kg bw of the filtrate corresponds to a dose of 2000 mg/kg bw.

Test animals

Species:

mouse

Strain:

NMRI

Sex:

male/female

Administration / exposure

Route of administration:

intraperitoneal

Vehicle:

distilled water

Duration of treatment / exposure:

Bone marrow cells were collected 24 h and 48 h (highest dose only) after dosing. Toxicity and thus exposure of the target cells was determined by measuring the ratio between polychromatic and total erythrocytes (PCE/TE). Bone marrow preparations were stained with May-Grunwald/Giemsa and examined microscopically for the NCE/TE ratio and micronuclei.

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

200 mg/kg bw (total dose)

Doses / concentrations 2

Dose / conc.:

1 000 mg/kg bw (total dose)

Doses / concentrations 3

Dose / conc.:

2 000 mg/kg bw (total dose)

Doses / concentrations 4

Dose / conc.:

0 mg/kg bw (total dose)

Control animals:

yes

Positive control(s):

Positive controls were in accordance with the OECD guideline.

Results and discussion

Test results

Sex:

not specified

Genotoxicity:

negative

Toxicity:

not specified

Vehicle controls validity:

not specified

Negative controls validity:

not specified

Positive controls validity:

not specified

Additional information on results:

In the pre-test for toxicity, mice treated with 2000 mg/kg bw showed systemic toxic effects, i.e. lethargy at 6, 24 and 48 h after dosing. Due to this result 2000 mg/kg bw was chosen as the top dose. In the main test no signs of toxicity were found within 24 h after dosing.
However, after 48 h 3 male mice died 1-2 h before bone marrow preparation whereas 1 male and 1 female mouse showed lethargy.
The bone marrow of the 3 dead mice was collected immediately after compound-related death. Due to the process of erythropoiesis in vivo and the mechanism of micronucleus formation in the polychromatic erythrocytes false positive or false negative results can be excluded. All other bone marrow preparations were performed according to the guideline.

Applicant's summary and conclusion

Conclusions:

Under the experimental conditions used 19556 Indigoblätter did not induce a biologically
relevant increase in the number of erythrocytes with micronuclei of treated mice and,
consequently, 19556 Indigoblätter is not genotoxic (clastogenic and/or aneugenic) in bone
marrow cells of mice.