File Name: ion chromatography principles and applications .zip
- 1 ION CHROMATOGRAPHY — PRINCIPLES AND APPLICATIONS
- Principles of Ion exchange chromatography
- Separation techniques: Chromatography
- Vol 31 No s4 LCGC North America April 2013 Advances in Ion Chromatography Supplement PDF
1 ION CHROMATOGRAPHY — PRINCIPLES AND APPLICATIONS
Chromatography is an important biophysical technique that enables the separation, identification, and purification of the components of a mixture for qualitative and quantitative analysis. Proteins can be purified based on characteristics such as size and shape, total charge, hydrophobic groups present on the surface, and binding capacity with the stationary phase. Four separation techniques based on molecular characteristics and interaction type use mechanisms of ion exchange, surface adsorption, partition, and size exclusion. Other chromatography techniques are based on the stationary bed, including column, thin layer, and paper chromatography. Column chromatography is one of the most common methods of protein purification.
Column Chromatography. Ion chromatography separation is based on ionic or electrostatic interactions between ionic and polar analytes, ions present in the eluent and ionic functional groups fixed to the chromatographic support. 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 chromatography is one of the most important adsorption techniques used in the separation of peptides, proteins, nucleic acids and related biopolymers which are charged molecules in different molecular sizes and molecular nature [ 3 - 6 ]. Biomolecules display different degrees of interaction with charged chromatography media due to their varying charge properties [ 8 ]. The earliest report of ion-exchange chromatography date back to , Thompson studied the adsorption of ammonium ions to soils [ 9 - 11 ].
Principles of Ion exchange chromatography
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Wenzhi Hu and Paul R. A new ion chromatographic method with unique separation selectivity and ultra-high sensitivity for the separation and detection of inorganic anions was developed. The method uses a conventional reversed-phase ODS stationary phase modified with a zwitterionic surfactant and an aqueous solution containing a small amount of electrolyte as the eluent. The elution order for the anions differed from that achieved by conventional anion-exchange, with for example, sulfate being eluted before chloride, and in contrast to the situation when water was used as mobile phase, only a single peak was observed for each analyte anion. Addition of the electrolyte to a water mobile phase resulted in an initial large increase in analyte retention times, but further increases in electrolyte concentration in the mobile phase produced only minor changes in retention time. Use of bicarbonate as the electrolyte anion permitted a suppressor to be used with conductometric detection. The ability to directly analyse samples of high ionic strength permitted the method to be applied to the determination of iodide in saline solution using direct UV absorbance detection, with a detection limit of 0.
Separation techniques: Chromatography
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The basic process of chromatography using ion exchange can be represented in 5 steps: eluent loading, sample injection, separation of sample, elution of analyte A, and elution of analyte B, shown and explained below. Elution is the process where the compound of interest is moved through the column. This happens because the eluent, the solution used as the solvent in chromatography, is constantly pumped through the column. The chemical reactions below are for an anion exchange process. Step 1 : The eluent loaded onto the column displaces any anions bonded to the resin and saturates the resin surface with the eluent anion.
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Vol 31 No s4 LCGC North America April 2013 Advances in Ion Chromatography Supplement PDF
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Ion chromatography or ion-exchange chromatography separates ions and polar molecules based on their affinity to the ion exchanger. It works on almost any kind of charged molecule —including large proteins , small nucleotides , and amino acids. However, ion chromatography must be done in conditions that are one unit away from the isoelectric point of a protein.
Such systems provide very good detectability, precision, and limits of quantifications. Ion chromatography can be used to determine: trace amounts of anions and.
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Ion Chromatography pp Cite as. Of the many methods that have been applied to the chromatographic separation of ions, most employ ion exchangers. The purpose of this chapter is to describe some of the more important phenomena associated with the ion exchange process, to explain their physical causes, and to demonstrate how principles and theories of ion exchange relate to the chromatographic separation of ions. Unable to display preview. Download preview PDF.
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