What is the principle of ion exchange chromatography?
Principle of Ion Exchange Chromatography The molecules separated on the basis of their charge are eluted using a solution of varying ionic strength. By passing such a solution through the column, highly selective separation of molecules according to their different charges takes place.
What is ion exchange chromatography PPT?
Introduction • “Ion exchange chromatography may be defined as the reversible exchange of ions in the solution with ions electrostatically bound to some sort of insoluble matrix or a stationary phase.” • This technique is extremely useful in the separation of charge compounds like proteins differing by only one charged …
What are the four steps of ion exchange chromatography?
Ion-exchange chromatography is generally a four-step process. First, a packed column containing either anion- or cation-exchange resin is equilibrated using buffer. For anion-exchange columns, this involves protonating the resin, ensuring it is positively charged. Next, the sample is loaded on the column.
What mobile phase is used in ion exchange chromatography?
In cation-exchange chromatography, the stationary phase, which consists of a large quantity of acid groups attached to a polymeric resin, is slurried with water and applied to a column. The mobile phase, which contains the inorganic salt dissolved in a suitable solvent, is applied to the column.
Why is NaCl used in ion exchange chromatography?
NaCl dissociates in Na+ and Cl- in solution. In anion-exchange chromatography, Cl- competes with negatively charged analytes to bind with positively charged stationary phase, thereby eluting the analytes.
What are the advantages of ion exchange chromatography?
Advantages of ion exchange chromatography t is one of the most efficient methods for the separation of charged particles. 2. It can be used for almost any kind of charged molecule including large proteins, small nucleotides and amino acids. 3.
What are the types of ion exchange?
Ion exchangers are either (i) cation exchangers, which exchange positively charged ions—cations, or (ii) anion exchangers which exchange negatively charged ions—anions.
Why is NaCl used in buffer?
Many buffers contain NaCl to help keep proteins soluble and to mimic physiological conditions. Generally, 150 mM NaCl is used. This will help screen ionic interactions and prevent nonspecific binding of proteins to the column while enabling your protein of interest to bind the column.
How does pH affect ion exchange chromatography?
The ion exchange chromatography matrix consists of positively and negatively charged ions. Essentially, molecules undergo electrostatic interactions with opposite charges on the stationary phase matrix. A change in pH affects the charge on the particular molecules and, therefore, alters binding.
What is the advantages of chromatography?
The Advantages of Chromatography Precise separation, analyses, and purification is possible using chromatography. It requires very low sample volumes. It works on a wide range of samples including drugs, food particles, plastics, pesticides, air and water samples, and tissue extracts.
What are the advantages and disadvantages of ion exchange?
Ion exchangers are quickly polluted, which considerably reduces the exchange capacity. Examples of this include pollution by micro-biology (e.g. film-forming bacteria) and pollution by suspended matter. Another disadvantage is the relatively high operational costs for, among other things, the regeneration fluid.
What are ion exchangers give example?
Ion exchangers are natural substances—for example, certain clays—or deliberately synthesized resins containing positive ions (cation exchangers) or negative ions (anion exchangers) that exchange with those ions in solution having a greater affinity for the exchanger.
What is ion exchange method?
Ion exchange is a method widely used in household (laundry detergents and water filters) to produce soft water. This is accomplished by exchanging calcium Ca 2+ and magnesium Mg 2+ cations against Na + or H + cations (see water softening).
What is ion exchange in chemistry?
ion exchange n (Chemistry) the process in which ions are exchanged between a solution and an insoluble solid, usually a resin. It is used to soften water, to separate radioactive isotopes, and to purify certain industrial chemicals.
What is ion exclusion chromatography?
Ion Exclusion. Ion exclusion chromatography is an alternative to ion exchange chromatography in which ionized samples are excluded from the pores of the support and elute first, while the weakly ionized and non-ionic compounds elute later.