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Toxicity to reproduction: other studies

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Administrative data

Endpoint:
toxicity to reproduction: other studies
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study meets basic scientific principles, well documented
Cross-reference
Reason / purpose:
reference to same study

Data source

Reference
Reference Type:
publication
Title:
Assessment of estrogenic activity in some common essential oil constituents
Author:
Howes M.-J.R., Houghton P.J., Barlow D.J., Pocock V.J., Milligan S.R.
Year:
2002
Bibliographic source:
J. Pharm. Pharmacol. 54, 1521-1528

Materials and methods

Principles of method if other than guideline:
Uterotrophic assay: uterus weights as indicator of estrogenic action after repeated exposure
Acute assay: vascular permeability of uterus as acute indicator of estrogenic action
GLP compliance:
not specified
Type of method:
in vivo

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Citral
- Analytical purity: 95%

Test animals

Species:
mouse
Strain:
Swiss
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: A. Tuck & Son Ltd., Essex, UK
- Age at study initiation: 3 mo
- Weight at study initiation: 25-35 g
- Fasting period before study:
- Diet: ad libitum
- Pretreatment: ovariectomy prerformed under anesthesia at least two weeks before the start of each experiment

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 +- 1
- Humidity (%): no data
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
dermal
Vehicle:
ethanol
Details on exposure:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): acute assay 100 µl; uterotrophic assay 50 µl
- Concentration (if solution): 1.9 M

VEHICLE
- Justification for use and choice of vehicle (if other than water): ethanol due to solubility

USE OF RESTRAINERS FOR PREVENTING INGESTION: no data
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
Acute assay of vascular responses: 4 h
Uterotrophic assay: 3 d
Frequency of treatment:
Acute assay: single
Uterotrophic assay: twice daily
Duration of test:
Acute assay: 4 h
Uterotrophic assay: sacrifice 12 h after last application
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
950 mg/kg bw
Basis:
other: calculated from dermal dose of 100 µl in the acute assay
Remarks:
Doses / Concentrations:
950 mg/kg bw/d
Basis:
other: calculated from two daily dermal doses of 50 µl in the uterotrophic assay
No. of animals per sex per dose:
5
Control animals:
yes, concurrent vehicle
other: positive control: 17ß-estradiol
Details on study design:
Acute assay of vascular responses:
4 hrs after the dermal application of the test substance, 50 μl 0.5 μCi [ 125 I]-labeled human serum albumin was injected into the jugular vein of mice anesthetised with tribromoethanol. Thirty minutes later, a blood sample was drawn from the suborbital canthal sinus with a heparinized capillary pipette and the animals were killed by cervical dislocation. The uteri and a sample of thigh muscle were removed, briefly washed in saline, and weighed. The radioactivity in the uterus, plasma sample, and muscle was determined. The tissue-specific extravascular albumin volume (EAV) was expressed as a ratio of the [ 125 I] counts per minute per milligram of tissue to [ 125 I] counts per minute per microliter of plasma and used as an index of tissue vascular permeability.

Uterotrophic assay:
12 hrs after the last dermal application, the mice were killed and the uterine horns removed, blotted and weighed.
Statistics:
Analysis of variance

Results and discussion

Any other information on results incl. tables

Table: Uterine weight (12 hrs after last application) and vascular permeability 4 hrs following transdermal application

Treatment

Uterine weight (mg)

Uterine vascular permeability (EAV)

Muscle vascular permeability (EAV)

Citral
950 mg/kg bw/d

12.5 ± 0.88

12.5 ± 4.31

0.95 ±- 0.04

Vehicle control

13.5 ± 0.28

5.96 ± 0.54

1.24 ± 0.04

Positive control
0.3 mM Estradiol

19.0 ± 0.15 *

20.3 ± 1.12 *

0.94 ± 0.03

significant change compared to vehicle control: * p<0.01

Applicant's summary and conclusion