Pressurized liquid extraction-assisted mussel cytosol preparation for the determination of metals bound to metallothionein-like proteins.
The possibilities of pressurized liquid extraction (PLE) have been novelty tested to assist the cytosol preparation from wet mussel soft tissue before the determination of metals bound to metallothionein-like proteins (MLPs). Results obtained after PLE were compared with those obtained after a classical blending procedure for mussel cytosolic preparation. Isoforms MLP-1 (retention time of 4.1 min) and MLP-2 (retention time of 7.4 min) were separated by anion exchange high-performance liquid chromatography (HPLC) and the concentrations of Ba, Cu, Mn, Sr and Zn bound to MLP isoforms were directly measured by inductively coupled plasma-atomic emission spectrometry (ICP-OES) as a multi-element detector. The optimized PLE-assisted mussel cytosol preparation has consisted of one extraction cycle at room temperature and 1500 psi for 2 min. Since separation between the solid mussel residue and the extract (cytosol) is performed by the PLE system, the cytosol preparation method is faster than conventional cytosol preparation methods by cutting/blending using Ultraturrax or Stomacher devices.
Santiago-Rivas
Sandra
S
Department of Analytical Chemistry, Nutrition and Bromatology, Faculty of Chemistry, University of Santiago de Compostela, Avenida das Ciencias s/n, 15782 Santiago de Compostela, Spain.
Moreda-Piñeiro
Antonio
A
Bermejo-Barrera
Pilar
P
Moreda-Piñeiro
Jorge
J
Alonso-Rodríguez
Elia
E
Muniategui-Lorenzo
Soledad
S
López-Mahía
Purificación
P
Prada-Rodríguez
Darío
D
D000818:Animals; D002852:Chromatography, Ion Exchange / Q000295:instrumentation / Q000379:methods; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism*; D008668:Metallothionein / Q000378:metabolism*; D019216:Metals, Heavy / Q000032:analysis* / Q000378:metabolism; D049878:Mytilus / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D011312:Pressure; D011485:Protein Binding; D014874:Water Pollutants, Chemical / Q000032:analysis* / Q000378:metabolism
D019216:Metals, Heavy; D014874:Water Pollutants, Chemical; D008668:Metallothionein