Abstract
Targeted quantification of protein biomarkers with liquid chromatography-tandem mass spectrometry (LC-MS/MS) has great potential, but is still in its infancy. Therefore, we elucidated the influence of charge state distribution and matrix effects on accurate quantification, illustrated by the peptide hormone hepcidin. An LC-MS/MS assay for hepcidin, developed based on existing literature, was improved by using 5 mM ammonium formate buffer as mobile phase A and as an elution solution for solid phase extraction (SPE) to optimize the charge state distribution. After extensive analytical validation, focusing on interference and matrix effects, the clinical consequence of this method adjustment was studied by performing receiving operating characteristic (ROC)-curve analysis in patients with iron deficiency anemia (IDA, n=44), anemia of chronic disease (ACD, n=42) and non-anemic patients (n=93). By using a buffered solution during sample preparation and chromatography, the most abundant charge state was shifted from 4+ to 3+ and the charge state distribution was strongly stabilized. The matrix effects which occurred in the 4+ state were therefore avoided, eliminating bias in the low concentration range of hepcidin. Consequently, sens...Continue Reading
References
Nov 16, 2002·Blood·Elizabeta NemethTomas Ganz
Oct 30, 2004·Science·Elizabeta NemethJerry Kaplan
Mar 16, 2005·Clinical Biochemistry·Paul J Taylor
Apr 17, 2007·Blood·Anthony T MurphyVictor J Wroblewski
Aug 12, 2008·Blood·Tomas GanzMark Westerman
Feb 25, 2009·PloS One·Vasiliki KoliarakiAvgi Mamalaki
Mar 5, 2009·Journal of Pharmacological and Toxicological Methods·Hongyan LiBarbra J Sasu
Dec 10, 2009·Haematologica·Joyce J C KrootDorine W Swinkels
Apr 15, 2010·Rapid Communications in Mass Spectrometry : RCM·Sukhvinder S BansalRobert C Hider
Sep 18, 2010·Clinical Chemistry·Anthony M ButterfieldRobert J Konrad
Nov 17, 2010·Rapid Communications in Mass Spectrometry : RCM·A K HewavitharanaP Kebarle
Nov 19, 2010·Bioanalysis·Eric Ezan, Francis Bitsch
Aug 27, 2011·Clinica Chimica Acta; International Journal of Clinical Chemistry·Sun-Il HwangHerbert L Bonkovsky
Oct 1, 2011·Analytical Chemistry·Damon S AndersonHanno Steen
Jan 10, 2012·Clinica Chimica Acta; International Journal of Clinical Chemistry·Outi ItkonenEsa Hämäläinen
Aug 14, 2012·American Journal of Hematology·Joyce J C KrootDorine W Swinkels
Jan 22, 2013·American Journal of Hematology·David B BregmanLawrence T Goodnough
May 7, 2013·Clinica Chimica Acta; International Journal of Clinical Chemistry·Fleur WolffFrédéric Cotton
Oct 22, 2013·Bioanalysis·Bertrand RochatSophie Waldvogel
Aug 27, 2014·Bioanalysis·Derek L ChappellOmar F Laterza
Mar 18, 2015·Clinical Chemistry and Laboratory Medicine : CCLM·Thibaud LefebvreZoubida Karim
Apr 22, 2015·PloS One·Martin WagnerLothar Schramm
Aug 22, 2015·Bioanalysis·Irene van den Broek, William D van Dongen
May 2, 2016·Clinical Biochemistry·Brian G Keevil
Oct 31, 2016·Analytical Biochemistry·Adam C SwensenJohn C Price
Aug 2, 2017·Endocrinology and Metabolism Clinics of North America·Siva S KethaHemamalini Ketha
Citations
Apr 25, 2019·Biochemia Medica·Dietmar EnkoMarkus Herrmann
Oct 2, 2020·Clinical Chemistry and Laboratory Medicine : CCLM·Ellis T AuneDorine W Swinkels
Oct 24, 2020·Mikrochimica Acta·Shilpa RanaNirmal Prabhakar
Jul 17, 2021·Journal of Proteome Research·Rachel E ForemanRichard G Kay