Novel HPLC-ICP-MS strategy for the determination of beta2-transferrin, the biomarker of cerebrospinal fluid (CSF) leakage.
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.
del Castillo Busto
Estela
E
Department of Physical and Analytical Chemistry, Faculty of Chemistry, University of Oviedo, Julián Clavería 8, 33006 Oviedo, Spain.
Montes-Bayón
Maria
M
García Alonso
J Ignacio
JI
Caruso
Joseph A
JA
Sanz-Medel
A
A
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*
D015415:Biomarkers; D007503:Iron Isotopes; D014168:Transferrin