Détermination de l’hémoglobine totale par dilution isotopique ICPMS afin d’assurer la traçabilité métrologique en biochimie clinique

Abstract

Abstract. The main objective of this study is the development of reference measurement procedures for the determination of total haemoglobin (Hb) in blood based by ID-ICPMS. First of all, a characterization of a Hb standard is carried out via total iron analysis by double ID-ICPMS. This standard will be used as a primary calibration solution in order to ensure the traceability of the measurement results. The validation of this method was conducted by the analysis of the CRM JCCRM 912-1 (ReCCS) certified in total Hb and taking into account the stoichiometry of the molecule (4Fe(Hb)). Furthermore, elemental speciation strategies are carried out through the use of hybrid techniques (e.g. HPLC-ICPMS). The separation of Hb is performed by liquid chromatography using cation or anion exchange columns. The detection is conducted by measuring the iron contained in the haeme-group of the protein, using a sector field ICPMS at medium resolution. In order to develop a primary reference method, isotope dilution analysis is carried out for the determination of total Hb by species-specific mode. For that purpose, the synthesis of an isotopically labelled standard Hb (Hb-57 Fe) is performed. This ID strategy is compared with the species-unspecific mode in terms of accuracy and precision.