Fractionation metallothionein-like proteins in mussels with on line metal detection by high performance liquid chromatography-inductively coupled plasma-optical emission spectrometry.
A rapid method for the determination of Al, Ba, Cu, Fe, Mn, Sr and Zn binding metallothionein-like proteins (MLPs) in mussels (Mytilus galloprovincialis) by inductively coupled plasma-optical emission spectrometry was developed. The method uses a short column (8mmx75mm) anion exchange high performance liquid chromatography (HPLC) with inductively coupled plasma-optical emission spectrometric (ICP-OES) detection. Working in isocratic mode (75mM Tris-HCl at pH 7.4, flow rate at 0.8mlmin(-1)), two major MLPs isoforms (MLP-1 and MLP-2) can be separated in ten minutes. The distribution of basal metals binding MLPs was assessed by on line HPLC and ICP-OES, while the basal contents of metals binding MLPs were obtained after off line HPLC and ICP-OES (collection of the two major fractions and multi-element determination by ICP-OES). The calculated LODs were 81, 6, 10, 13, 30, 9 and 123ngg(-1) for Al, Ba, Cu, Fe, Mn, Sr and Zn binding MLP-1 and MLP-2, respectively. The repeatability of the over-all method (five different mussel cytosols prepared from the same mussel sample and subjected to the off line HPLC-ICP-OES procedure twice) was from 11.2 for Cu to 16.2% for Zn. The method was finally applied to different raft mussels from Ría de Arousa estuary in order to know basal levels of elements binding MLPs.
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
Adela
A
Bermejo-Barrera
Pilar
P