Speciation of Trace Elements, Binding and Non-binding with Proteins in Human Blood Serum, by Surfactant-Mediated HPLC with Element-Selective Detection by ICP-MS

摘要: The total concentrations of some trace elements in blood serum are generally used as convenient parameters medical diagnosis because their extreme importance for health and disease,1 well easy accessibility. However, most the combined with biological substances to have own specific functions. Therefore, more information about chemical species elements, addition concentrations,2,3 is required investigations on physiological functions biomedical physical conditions. Chemical divided into two forms. 2 One protein-associated species, which play important roles enzymatic activities, transportation storage elements. other protein-non-associated related membrane transport or excretion In a previous work, 4 present authors developed surfactant-mediated chromatographic separation system allowed simultaneous multi-component separation, based mixed-mode properties such size-exclusion, electrostatic hydrophobic interactions. this system, large-molecule (MW 6500‐2000000) were eluted within 3 min elution time, while small-molecule (MW< 1000) after 3.5 min. Thus, could be direct injection analysis samples separate drugs rapidly from proteins human without any pretreatment, where UV absorption detector was employed. Since column also allows us couple inductively coupled plasma mass spectrometry (ICP-MS) multielement detection, HPLC/ICP-MS using can applied speciation sample injection. paper, we report preliminary study binding non-binding carried out by system. HPLC composed pump (Model LC10AD; Shimadzu, Kyoto), injector V7, Pharmacia-LKB, Uppsala, Sweden) 20 µl loop, 870-UV; Jasco, Tokyo). detection UV-absorbing species. experiment, an ODS (L-column; 4.6 mm i. d. × 250 long; Inspection Testing Institute, Tokyo) dynamically coated 3-[(3-cholamidopropyl)dimethylammonium]-1-propanesulfonate (CHAPS), zwitterionic bile acid derivative, employed column. CHAPS-coated prepared manner similar that paper. mobile phase 0.2 mM Tris‐HCl buffer solution (pH 7.4) at flow rate 0.7 ml ‐1 , CHAPS added prevent degradation For on-line element-selective ICP-MS instrument SPQ8000A, Seiko Instruments, Chiba) introduction eluent nebulizer via Teflon tubing. operated time-sequential program, chromatogram measurements. obtained healthy volunteers. sampling pretreatment procedures performed those 5 diverse physicochemical variety solutes, inorganic anions, amino acids, aromatic compounds,