Ion chromatography or Ion-exchange chromatography is a procedure which enables the separation of ions and polar molecules on the basis of the charge properties of these molecules. It may be used for many types of atoms including large proteins, little nucleotides and amino acids. The solution to be injected is normally known as a sample and the independently separated elements are identified as analyses. It is often utilized in protein purification, water analysis and for quality control purposes.
Ion exchange chromatography Keeps analyze molecules with columbic ionic interactions. The stationary phase surface exhibits ionic functional groups that interact with analyte ions of opposite charge. This category of chromatography can be further subdivided into action exchange chromatography and anion exchange chromatography. The ionic compound comprising the cationic species and the anionic species may be retained by the stationary phase.
Action exchange ion Chromatography retains positively charged captions because the stationary phase exhibits a negatively charged functional class. Anion exchange chromatogram keeps anions displaying a positively charged functional class. Note that the ion power of captions or anions in the mobile phase may be adjusted to alter the balance position and, hence, the retention period. An ion chromatogram can be used to reveal that the chromatogram obtained with an ion exchange column.
A Normal ion chromatography Technique involves the introduction of a sample either manually or using an auto sampler, to a sample loop of known quantity. A buffered aqueous solution called the mobile phase carries the sample out of the loop into a column that comprises some kind of stationary phase material. This is normally a resin or gel matrix which includes agarose or cellulose beads with covalently bonded charged functional groups. The target analytes anions or cations are kept on the stationary phase but may be eluted by increasing the concentration of a similarly charged species. This will displace the analyte ions in the static phase.
For Instance, in cation exchange chromatography, the positively charged analyte could be displaced by the introduction of positively charged sodium ions. The analytes of interest must then be discovered by some procedure, which is normally by either conductivity or UV/Visible light absorbance. So as to control an ion Chromatography system, a chromatography data system is usually required. Some of the chromatography data systems are also used to control gas chromatography and HPLC systems. Proteins have many functional Groups that could have both negative and positive charges.
Ion chromatography separates Proteins based on their net charge. This depends upon the composition of the mobile phase. By adjusting the pH or the ionic concentration of the mobile phase, various protein molecules can be separated. By way of instance, if a protein has a net positive charge at pH 7, then it will bind to a column of negatively-charged beads, but a negatively charged protein would not. Changing the pH so the net charge on the protein is negative will make it also be eluted.