title: Validated HPLC assay for iron determination in biological matrices based on ferrioxamine formation.
abstract: A simple, robust and reproducible HPLC method has been developed and validated for iron determination in biological matrices. It is based on chelation with desferrioxamine (DFO) and the measurement of the chelate ferrioxamine (FO). The method was developed to permit monitoring of iron bio-kinetics and estimation of iron status in experimental animals. The chromatography was performed on a stainless steel XTerra MS C18 column (Waters; 250 mm x 4.6 mm i.d., 5 microm) using a gradient of Tris-HCl buffer (10mM, pH 5) and acetonitrile. The method was validated in terms of selectivity, linearity (0.3-80 nmol on-column), limit of detection (0.2 nmol on-column), low limit of quantification (0.3 nmol on-column), recovery (91-102%), intra- and inter-day reproducibility, stability, and robustness. The method's universal applicability was illustrated by monitoring plasma and heart iron kinetic profiles in rats after a single intraperitoneal (i.p.) injection of 200mg/kg iron dextran.
authors: Leslie Dan Faculty of Pharmacy, University of Toronto, 19 Russell St., Toronto, Ont., Canada M5S 2S2.
authors: Angelo
authors: A
authors: Tesoro
authors: Jasmina
authors: J
authors: Novakovic
authors: Jake J
authors: JJ
authors: Thiessen
authors: Michael
authors: M
authors: Spino
mesh_terms: D000818:Animals; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003676:Deferoxamine / Q000032:analysis* / Q000737:chemistry; D005290:Ferric Compounds / Q000032:analysis*; D007274:Injections, Intraperitoneal; D007501:Iron / Q000032:analysis* / Q000097:blood / Q000493:pharmacokinetics; D007502:Iron Chelating Agents / Q000737:chemistry; D007700:Kinetics; D008297:Male; D009206:Myocardium / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results
chemical_list: D005290:Ferric Compounds; D007502:Iron Chelating Agents; C002577:ferrioxamine B; D007501:Iron; D003676:Deferoxamine