Determination of Non-Transferrin Bound Iron, Transferrin Bound Iron, Drug Bound Iron and Total Iron in Serum in a Rats after IV Administration of Sodium Ferric Gluconate Complex by Simple Ultrafiltration Inductively Coupled Plasma Mass Spectrometric Detection

Abstract

A rapid, sensitive and specific ultrafiltration inductively-coupled plasma mass spectrometry method was developed and validated for the quantification of non-transferrin bound iron (NTBI), transferrin bound iron (TBI), drug bound iron (DI) and total iron (TI) in the same rat serum sample after intravenous (IV) administration of iron gluconate nanoparticles in sucrose solution (Ferrlecit®). Ultrafiltration with a 30 kDa molecular cut-off filter was used for sample cleanup. Different elution solvents were used to separate each form of iron from sample serum. Isolated fractions were subjected to inductively-coupled mass spectrometric analysis after microwave digestion in 4% nitric acid. The reproducibility of the method was evaluated by precision and accuracy. The calibration curve demonstrated linearity from 5–500 ng/mL with a regression (r2) of more than 0.998. This method was effectively implemented to quantify rat pharmacokinetic study samples after intravenous administration of Ferrlecit®. The method was successfully applied to a pharmacokinetic (PK) study of Ferrlecit in rats. The colloidal iron followed first order kinetics with half-life of 2.2 h and reached background or pre-dose levels after 12 h post-dosing. The drug shown a clearance of 0.31 mL/min/kg and volume of distribution of 0.05 L/kg. 19.4 ± 2.4 mL/h/kg.

Comparative Evaluation of U.S. Brand and Generic Intravenous Sodium Ferric Gluconate Complex in Sucrose Injection: Biodistribution after Intravenous Dosing in Rats

“…The overall tissue distribution of the two drugs was very similar. The early time point kinetics of both SFG drugs in serum were similar to those observed previously in the pharmacokinetic study [ 14 ]. At some time points, differences in product related iron concentration was observed in one or two tissues.…”

“…Spurred by discrepancies between FDA’s 2013 Draft Guidance on Sodium Ferric Gluconate Complex [ 9 ] and the EMA’s 2011 Draft Reflection Paper on the Data Requirements for Intravenous Iron-based Nano-colloidal Products Developed with Reference to an Innovator Medicinal Product [ 11 ], the two FDA approved SFG products were investigated in a comprehensive BD study, confirming no significant and biologically relevant differences. Prior to initiating this BD study, product physiochemical characterization [ 12 ], cellular uptake [ 13 ], and pharmacokinetic [ 14 ] studies were completed to inform the BD study design. The BD study incorporated serum and bone marrow to monitor pharmacological delivery and target tissues.…”

“…Prior to design of the biodistribution study, a pharmacokinetic (PK) study [ 14 ] was conducted with the innovator product that demonstrated a returned to baseline by 24 h after dosing. Based on that PK study and the recognized tissue retention of iron from intravenous iron oxide nanoparticle products [ 16 , 17 ], immediate time points (<24 h) were included to assure that similar dosing and PK parameters were obtained, that free iron of potential toxicity was not different and that immediate organ distribution was the same between the innovator and generic.…”

“…The results of these analyses are reported in a separate publication that describes, in detail, the minor differences detected between the products and interprets those differences in the context of a generic product review [ 12 , 13 ]. A pharmacokinetic study [ 14 ] was conducted to determine early time points and a dose concentration for the BD study. The BD study organs included blood serum to measure maximal drug iron exposure and its rate of systemic delivery, bone marrow as the designated pharmacological target tissue, lungs, brain, and heart as critical organs of toxicity concern, and spleen, liver, and kidney as the primary organs of uptake, storage, and excretion.…”

Alteration of Iron Concentration in Alzheimer’s Disease as a Possible Diagnostic Biomarker Unveiling Ferroptosis

“…[147] S Chemistry analyzers, i.e., colorimetric reaction with ferrine or ferrozine as a chromogen to form a color complex with iron. [149]; HPLC [148]; isotope dilution MS [150]; novel NTBI measuring system using nitrilotriacetate and PSAP as chromogen [151]; chelatable fluorescent beads based on flow cytometry [152]; fluorescent bead assay CP851 chelator based [153].…”