Mangafodipir trisodium injection, a new contrast medium for magnetic resonance imaging: detection and quantitation of the parent compound MnDPDP and metabolites in human plasma by high performance liquid chromatography

Abstract

Manganese(II) dipyridoxyl diphosphate (MnDPDP) is the active component of mangafodipir trisodium injection (Teslascan ®), a new contrast medium for magnetic resonance imaging. A high performance liquid chromatography (HPLC) method was developed for the simultaneous determination of MnDPDP and its five major metabolites in human plasma, i.e. the dephosphorylation products MnDPMP (manganese(II) dipyridoxyl monophosphate) and MnPLED (manganese(II) dipyridoxyl ethylenediamine diacetate) and the corresponding substances obtained after transmetallation with zinc (ZnDPDP, ZnDPMP and ZnPLED). Heparinized blood samples from patients receiving mangafodipir trisodium injection were immediately mixed with solid trisodium phosphate dodecahydrate to obtain pH 10.0 ± 0.2 in order to inhibit further in vitro dephosphorylation and transmetallation. The plasma thus obtained was ultrafiltrated prior to HPLC analysis. The chromatographic separation was obtained using a mixed-bed resin with both anion exchange and reversed-phase functions (OmniPac ™ PAX-500) using isocratic elution and UV detection at 310 nm. With an injection volume of 50 μl, the limit of quantitation (LOQ) values were 0.8–2.3 μM for the Mn compounds and 0.1–0.8 μM for the Zn compounds. The between-run accuracy of spiked plasma samples was in the range 97.5–106.7%, with a precision in the range 3.1–9.0%. The best fit calibration curves were obtained using non-linear regression according to the equation Y = A + BX M in the concentration range from LOQ to 100 μM. Long-term storage of spiked plasma samples for three months at − 20°C demonstrated the required stability with recovery values within 85–115% of MnDPDP and its five metabolites.