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

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-57Fe) is performed. This ID strategy is compared with the species-unspecific mode in terms of accuracy and precision.