Registration Dossier

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2002
Report Date:
2002

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
GLP compliance:
yes (incl. certificate)
Type of assay:
micronucleus assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Tridecylamine isomers (CAS 86089-17-0)
- Substance no.
00/0693-1
- Purity 99.6%
- Batch No.: Tank 66 v. 16.11.00
- Appearance, consistency: Colorless liquid
- Storage: Room temperature (N2 conditions)
- Stability: proven by reanalysis

Test animals

Species:
mouse
Strain:
other: Crl:NMRI
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland GmbH
- Age at study initiation: 5 - 8 weeks
- Weight at study initiation: mean ca. 28 g
- Housing: single housing in Makrolon cages, type Ml
- Diet: Kliba Haltungsdiät (Provimi Kliba SA, Kaiseraugst, Switzerland) ad libitum
- Water: ad libitum
- Acclimation period: at least 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12 / 12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
- Vehicle: olive oil Ph.Eur./DAB
- Justification for choice of vehicle: Due to the insolubility of the test substance in water, olive oil was selected as the vehicle, which had been demonstrated to be suitable in the in vivo micronucleus test and for which historical data are available.
- Concentration of test material in vehicle: 2, 4 or 6 g/100 ml
Details on exposure:
APPLIED VOLUME:
10 ml/kg bw
Duration of treatment / exposure:
- Sacrifice intervals: 24 hours (vehicle control, 200, 400 and 600 mg/kg TS and positive controls) and 48 hours (vehicle control and 600 mg/kg TS)
Frequency of treatment:
single oral administration
Doses / concentrations
Remarks:
Doses / Concentrations:
200, 400, 600 mg/kg
Basis:

No. of animals per sex per dose:
5 males
Control animals:
yes, concurrent vehicle
Positive control(s):
Cyclophosphamide (CPP) and Vincristine Sulphate (VCR)
- Justification for choice of positive control: The stability of CPP and VCR is well-defined under the selected conditions, since both positive control articles are well-defined clastogens and aneugens, respectively.
- Route of administration: oral by gavage
- Doses / concentrations: CPP: 20 mg/kg bw, VCR: 0.15 mg/kg bw

Examinations

Tissues and cell types examined:
Erythrocytes were prepared from femur bone marrow and stained for microscopic evaluation. 2000 polychromatic erythrocytes per animal were evaluated and the following parameters were recorded: number of polychromatic and normochromatic erythrocytes, each with and without micronuclei; number of small and large micronuclei. The ratio of polychromatic to normochromatic erythrocytes was calculated from the recorded data.
Evaluation criteria:
The test is considered valid and acceptable if the slides allow to evaluate sufficient cells, i.e. >/= 2000 polychromatic cells; and if the proportion of micronuclei in negative controls was within the range of historical control data; and if the two positive control chemicals induced an increase within the range of historical positive control data. The test chemical is considered positive if a dose-related and significant increase in the number of micronucleated polychromatic erythrocytes is noted at any of the intervals; and if the proportion of micronuclei bearing cells exceeded both the concurrent and the historical negative control range. If these criteria are not met the test chemical is considered negative in this test.
Statistics:
The Wilcoxon test for the hypothesis of equal medians was used to compare dose groups with vehicle groups.

Results and discussion

Test results
Sex:
male
Genotoxicity:
negative
Vehicle controls valid:
yes
Positive controls valid:
yes
Additional information on results:
CLINICAL SIGNS
In the pretest, mortalities were noted down to 1000 mg/kg body weight. 800 mg/kg were survived by both male and female mice, but caused tremor, squatting posture, and poor general state. In the main test, 5/10 animals died at 800 mg/kg within 48 h. The highest dose was therefore lowered to 600 mg/kg. No clinical signs were noted at 200 mg/kg. Transient squatting posture was seen in animals receiving 400 mg/kg 1 h after dosing. At 600 mg/kg, squatting posture was seen in all animals on the first and second day after treatment as was tremor. Poor general state was seen 1 h after treatment in all high dose animals.


ERYTHROCYTES
In the negative control groups, polychromatic erythrocytes containing micronuclei amounted to 1.0 ‰ after 24 h and 0.7 ‰ after 48 h. Cyclophosphamide led to exclusively small micronuclei, incidence was 14.8 ‰. Proportion following vincristine treatment was 99.6 ‰, with an expected amount of large micronuclei, i.e. 27.8 ‰. Effects of both positive control chemicals were statistically significant on the P<0.01 level.
Single doses of Tridecylamine led to 0.8 ‰ (200 mg/kg), 0.9 ‰ (400 mg/kg). The high dose led to 2.2 ‰ (24 h) and 1.1 ‰ (48 h). This was within the range of historical control data.
The number of normochromatic erythrocytes containing micronuclei was unchanged in the various dose groups. The number of normochromatic or polychromatic erythrocytes containing small micronuclei (d < D/4) or large micronuclei (d > D/4) did not deviate from the vehicle control value at any of the sacrifice intervals and was within the historical control range. A slight inhibition of erythropoiesis was seen at 600 mg/kg, as determined from the ratio of polychromatic to normochromatic erythrocytes.

Any other information on results incl. tables

Ratio polychromatic (PCE) and normochromatic (NCE) erythrocytes:

Dose (mg/kg bw)

Interval (h)

Ratio PCE/NCE

0

24

3.11

0

48

2.82

200

24

3.36

400

24

3.03

600

24

2.16

600

48

2.13

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): negative
Tridecylamine was negative in the mouse micronucleus, as the
proportion of micronuclei in polychromatic erythrocytes was
comparable to the controls or within the range of historical
control data (mean ± SD: 2.0±0.6 ‰; n=56) for olive oil.

The compound was tested up to dose levels which exerted
signs of toxicity. At the highest dose level, 600 mg/kg,
slight inhibition of erythropoiesis was noted. Thus the
target was reached by test chemical.

Overall, there was no indication of a clastogenic or
aneugenic activity.