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Two distinct mechanisms as follows; ion exchange due to competitive ionic binding (attraction) and ion exclusion due to repulsion between similarly charged analyte ions and the ions fixed on the chromatographic support, play a role in the separation in ion chromatography. Ion exchange has been the predominant form of ion chromatography to date [2]. This chromatog‐ raphy is one of the most Ion exchange chromatography involves the separation of ionizable molecules based on their total charge. This technique enables the separation of similar types of molecules that would be difficult to separate by other techniques because the charge carried by the molecule of interest can be readily manipulated by changing buffer pH. Depending on the pH of their environment, proteins may carry a IEX : Chromatographie échangeuse d'ions / Ion Exchange Chromatographie. CIEX :CEX :Cation Exchange. AIES :AEX :Anion Exchange Chromato. La séparation en IEX dépend de l'adsorption réversible de molé chargé sur des groupement échangeurs d'ions immobilité de charges opposé (contre-ion). Dans l'échange d'ions, les ions liés par interaction électrostatique (liaison ionique • • hplcis a separation technique that involves: • •the injection of a small volume of liquid sample into a tube packed with tiny particles (3 to 5 micron (μm) in diameter called the stationary phase) • •where individual components of the sample are moved down the packed tube (column) with a liquid (mobile phase) forced through the column by high … 26 separation column. In HPLC, the mobile phase (liquid) pressure, flow rate, linear 27 velocity, and the polarity of the mobile phase all affect a compounds' retention 28 time. An illustration of retention time is shown in Figure 1.2. The equation at the 29 top of the figure will be discussed later during our mathematic development of The anion exchanger Nucleosil SB is used for ion exchange chromatography. It is a strongly basic anion exchanger with a quaternary ammonium modification and a capacity of about 1 meq/g. . E150NS050Nucleosil 100-5-SB SAX 5 µm spherical 100 Å 350 m2/g 10 no MACHEREY-NAGEL . differences in their specific properties, as shown in Figure I.1. Ion exchange chromatography (IEX) separates biomolecules according to differences in their net surface charge. Property Technique Charge Ion exchange chromatography (IEX) Size Size exclusion chromatography (SEC), also called gel filtration (GF) Intro to Ion Chromatography (IC) •HPLC - Neutral samples (last Thu.) - Ionic samples • Inorganic ions • Ionized or ionizable organic molecules •IC - More complicated than neutral HPLC - Also some special problems - But more control is available • Easier to achieve successful separation. HPLC Approaches to Ionic Samples? • Reversed-Phase IC • Ion-Pair C • Ion-Exchange Both anion-exchange (AEX) (2,3,5,8,32,36 - 40) and cation-exchange (CEX) (2,3,5,8,31,32,34,36,38,41 - 45) HPLC have been employed for peptide and protein separations, negatively charged and positively charged solutes, respectively, being retained by these ion-exchange modes. Common anion-exchange packings consist of primary, secondary, and tertiary (weak AEX) or quaternary amine (strong 1 Introduction. Ion exc