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Toxicological information

Basic toxicokinetics

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Administrative data

Endpoint:
basic toxicokinetics in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: well documented study, GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2011
Report Date:
2012

Materials and methods

Test guideline
Qualifier:
no guideline followed
Principles of method if other than guideline:
The aim of this study was to compare the in vitro metabolism by hepatocytes of five test substances between 4 species (mouse, rat, rabbit and human). The analytical method utilised HPLC coupled with mass spectrometry (LC MS) to profile and identify metabolites generated.
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
- Name of test material (as cited in study report): anisaldehyde, p-methoxybenzaldehyde, anisic aldehyde
- Supplier: Charles River Laboratories (manufactured by BASF)
- Physical state: Liquid, colourless to slight yellow
- Analytical purity: 99.9%
- Batch No.: 00020777L0
- Expiration date of the batch: 31 October 2011
- Storage condition of test material: Room temperature, protected from light, in a tightly closed container (in a dry and well ventilated place)
Radiolabelling:
no

Test animals

Species:
other: not applicable (in vitro test)
Strain:
other: not applicable (in vitro test)

Administration / exposure

Route of administration:
other: not applicable (in vitro test)
Vehicle:
ethanol
Duration and frequency of treatment / exposure:
0, 1 or 4 h
Doses / concentrations
Remarks:
Doses / Concentrations:
1, 10 and 100 µM
No. of animals per sex per dose:
not applicable (in vitro test)
Control animals:
other: not applicable (in vitro test)

Results and discussion

Main ADME results
Type:
metabolism

Metabolite characterisation studies

Metabolites identified:
yes
Details on metabolites:
Following hepatocyte incubations of Anisaldehyde, a total of seven components were observed (see Annex 1):
A4: Anisic acid
A2: Anisic acid - glycine conjugate
A3: Anisic alcohol - glucuronide conjugate
A1: demethylated Anisic alcohol - glucuronide conjugate
A5-7: three hydroxylated Anisaldehyde - glucuronide conjugates

Metabolite A4 (anisic acid) was the only metabolite identified in the samples of the 0h incubation timepoint.
Metabolite A2 (glycine conjugate of Anisic acid) was the largest component in most 1 and 4 h incubations.
For rat hepatocyte incubations, metabolite A3 (glucuronide conjugate of Anisic alcohol) was generally the second largest component, whilst for the other species, metabolite A4 (Anisic acid) was typically the second largest component in the 1 and 4h incubations. The remaining metabolites of Anisaldehyde were typically observed at low levels and/or in a limited number of incubations. Unchanged anisaldehyde was not detectable by mass spectrometry.

Besides the predominant formation of Anisic alcohol - glucuronide conjugate in rats and to a lesser extent in the mouse, interspecies differences in the metabolite profiles were generally small.

Any other information on results incl. tables

The metabolic capacity of the hepatocytes used was assessed using 7 -ethoxy[14C]coumarin as standart substrate. Changes in 7-EC levels indicated, that all hepatocytes used were viable and capable of metabolism under the chosen testing conditions.

Applicant's summary and conclusion