3. FOOD ChEMISTRy & bIOTEChNOLOGy 3.1. Lectures

Chem. Listy, 102, s265–s1311 (2008) Food Chemistry & Biotechnology
to the extracts, they were incubated for 30 min at room temperature
and, finally, were centrifuged at 80,000 rpm for 1 h.
Extracts were stored at –80 °C until analysis.
Results
The profiles of different elements, such as Cu, Pb, Zn, ni,
As, Mn were obtained with two SEC: High molecular mass
(HMM) for a separation range of 3–70 kDa and low molecular
mass (LMM) for a separation range below 10 kDa ref. 3 .
This work focuses the profile of copper from different
organs with the LMM column. In this figure, a differential
peak between 2,126 to 7,000 Da was observed in the brain
extract.
Fig. 1. Molecular size distribution patterns of Cu obtained by
SEC-ICP-MS with the LMw column
For this reason, the copper fraction from brain was isolated
for a preliminary study and was used a second chromatographic
separation to purify the fraction previously mentioned
by using (RP)-HPLC and ICP-MS.
In Fig. 2., we can observe the two peaks obtained.
The mass balance corresponding to both chromatography
separation of this element was performed. A good recovery
is obtained which de notes an acceptable isolation of the
Cu molecules. For SEC the recovery of cooper from the total
injected was 44.40 % and for (RP)-HPLC 76.5 %. The first
peak contains 44.43 % of cooper from the total recovery for
(RP)-HPLC and the second one 32.07 %. The total concentrations
of cooper have been reported elsewhere 3 .
Conclusions
Previous works report the molecular size distribution
patterns of elements in Mus musculus mice. In spite of Mus
musculus is well known since a great number of proteomic
s640
Fig. 2. Elution profiles of Cu by (RP)-HPCL-ICPMS from the
brain extract
and genomic studies have been performed (i.e., the genome is
completely known), the metallome has not been reported until
now. The most interesting fraction is that from Cu in brain
extract which is not present in any other organ. This fraction
has been latterly purified by RP-HPLC-ICP-MS. The pure
extracts from RP-HPLC present only two peaks that can be
related to Cu-biomolecules. Future works will be focused
in the identification of these copper-compounds by organic
mass spectrometry.
The ICP-MS constitutes a powerful technique to obtain
the patterns of many elements in complex biological tissues
and it is an excellent guide for further studies of the fraction
of interest like further separations by orthogonal techniques,
molecular mass-spectrometric elucidations of the metallospecies
involved and so on.
This work has been supported by the projects CTM2006-
08960-C01/02 and BFU2005-02896 from the Ministerio de
Educación y Ciencia and by projects FQM-348 and RNM-
523 from the Consejería de Innovación, Ciencia y Empresa
(Junta de Andalucía). M.G.-F. thanks the Ministerio de Educación
y Ciencia for a predoctoral scholarship.
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