title: Novel HPLC-ICP-MS strategy for the determination of beta2-transferrin, the biomarker of cerebrospinal fluid (CSF) leakage.
abstract: Asialo-Transferrin (S(0)) or beta(2)-Transferrin is an accepted marker of cerebrospinal fluid (CSF) leakage from the subarachnoid space into the nasal or aural cavity as the result of a head trauma. At this time, the S(0) detection method of choice is isoelectric focusing on polyacrylamide gel with direct immunofixation of transferrin (Tf) and silverstaining which are time-consuming techniques (> 5 h). The aim of this study is to present an alternative methodology for determination of Tf sialoforms present in CSF samples, specifically S(0), based on the detection of the Fe associated with those forms. A double spiking methodology is developed by saturating the protein with isotopically enriched iron ((57)Fe) and conducting post-column Isotope Dilution Analysis (IDA) with (54)Fe after separation of the different sialoforms by anion exchange chromatography. The results obtained have been validated using a certified serum (ERM-DA470) and applied with satisfactory results to a pooled CSF sample.
authors.affiliation: Department of Physical and Analytical Chemistry, Faculty of Chemistry, University of Oviedo, Julián Clavería 8, 33006 Oviedo, Spain.
authors.forename: Estela
authors.initials: E
authors.lastname: del Castillo Busto
authors.forename: Maria
authors.initials: M
authors.lastname: Montes-Bayón
authors.forename: J Ignacio
authors.initials: JI
authors.lastname: García Alonso
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Caruso
authors.forename: A
authors.initials: A
authors.lastname: Sanz-Medel
mesh_terms: D015415:Biomarkers / Q000134:cerebrospinal fluid; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006259:Craniocerebral Trauma / Q000175:diagnosis; D006801:Humans; D007503:Iron Isotopes / Q000737:chemistry; D007553:Isotope Labeling; D013058:Mass Spectrometry / Q000379:methods*; D014168:Transferrin / Q000134:cerebrospinal fluid*
chemical_list: D015415:Biomarkers; D007503:Iron Isotopes; D014168:Transferrin