ION EXCHANGE CHROMATOGRAPHY

CHROMATOGRAPHY
The word chromatography literally means color writing Chromatography is a technique for separating mixtures into their components in order to analyze, identify, purify, and/or quantify the mixture or components.

Chromatography is used by scientists to:

Analyze examine a mixture, its components, and their relations to one another

Identify determine the identity of a mixture or

components based on known components

Purify separate components in order to isolate one

of interest for further study

Quantify determine the amount of the a mixture

and/or the components present in the sample

Components of Chromatography
1) Matrix (or stationary phase) usually an inert solid or gel and may be associated with various moieties, which interact with the analyte(s) of interest. 2) Mobile phase usually a liquid or a gas, is used to transport the analytes through the stationary phase.

 Ion Exchange Chromatography relies on the reversible exchange of ions in solution with ions electrostatically bound to an insoluble support media. There can be two types of functional groups covalently attached to the support beads. These are called 1) Anion exchangers (resin with positive functional groups) - negatively charged molecules are attracted to a positively charged solid support. 2) Cation exchangers (resin with negative functional groups) - positively charged molecules are attracted to a negatively charged solid support.

PRINCIPLE:

ION EXCHANGE RESIN

An ion exchange resin or ion exchange polymer is an insoluble matrix (or support structure) normally in the form of small beads, usually white or yellowish, fabricated from an organic polymer substrates. The material has highly developed structure of pores on the surface which are sites with easily trapped and released ions.

 The trapping of ions takes place only with simultaneous releasing of other ions; thus the process is called ION EXCHANGE.

SELECTIVITY/AFFINITY OF IONS
The degree to which the exchange takes place is limited by the preference the resin exhibits for the ion in solution. Consequently, the use of the resins exchange capacity will be limited unless the selectivity for the ion in solution is far greater than for the exchangeable ion attached to the resin. Generally, ions with higher valence, greater atomic weights and smaller radii are said to have a greater affinity for (be preferred by) ION EXCHANGE resins. Relative affinities of common ions are:

Ag+ > Cs+ > K+ > Na+ > Li+ Ba+2 > Sr+2 > Ca+2 > Mg+2 I- > NO3- > CN- -> HSO4- > NO2- > Cl- >HCO3-

Those substances with high affinities can continue to load to higher concentrations on the resins by displacing other previously exchanged potentially regulated ions with lower relative affinities. This is referred as CHROMATOGRAPHIC PEAKING.

RESIN REGENERATION
When the capacity of the resin is exhausted, it is necessary to regenerate the resin using a saturated solution to restore the capacity of the resin and return the resin to its initial condition. Brine, or sodium chloride solution, is most the commonly used regenerant, although others, such as strong acids (hydrochloric acid, sulfuric acid) or strong bases (sodium hydroxide) may also be used.

 Regeneration Procedure. After the feed solution is processed to the extent that the resin becomes exhausted and cannot accomplish any further ion exchange, the resin must be regenerated., Regeneration employs the following basic steps: 1. The column is backwashed to remove suspended solids collected by the bed during the service cycle and to eliminate channels that may have formed during this cycle. The back- wash flow fluidizes the bed. releases trapped particles. and reorients the resin particles according to size.

During backwash the larger, denser particles will accumulate at the base and the particle size will decrease moving up the column. This distribution yields a good hydraulic flow pattern and resistance to fouling by suspended solids.

2) The resin bed is brought in contact with the regenerant solution. 3) The resin bed is subjected to a fast rinse that removes the last traces of the regenerant solution and ensures good flow characteristics. 4) The column is returned to service.

BATCH AND COLUMN EXCHANGE SYSTEMS

Ion exchange processing can be accomplished by either a BATCH METHOD or a COLUMN/CONTINUOUS METHOD. 1) BATCH METHOD The resin and solution are mixed in a batch tank, the exchange is allowed to come to equilibrium, then the resin is separated from solution Fundamental concept involved is Chemical Equilibrium k (assign: Recall calculations involving chemical equilibrium)

2) CONTINUOUS/COLUMN METHOD Continuous ion exchange processes are usually of the down-flow and packed-bed column type Passing a solution through a column containing a bed of exchange resin is analogous to treating the solution in an infinite series of batch tanks

 The primary residual generated by ion exchange processes is the SPENT REGENERANT. The spent regenerant will have very high total dissolved solids (TDS) concentrations, as it will include all of the ions removed by the resin as well as the excess regenerant ions

In water treatment, it is primarily used for softening where calcium and magnesium ions are removed from water

PAPER CHROMATOGRAPHY

 Paper chromatography is the technique in which the separation of an unknown substance is mainly carried out by the flow of solvents on the specially designed chromatographic paper. Paper chromatography is an analytical method technique for separating and identifying mixtures that are or can be colored, especially pigments.

A few categories of pigments are listed below along with their characteristic range of colors.

Some plant pigments you may be familiar with that are of current interest in nutritional and pharmaceutical research are listed below

 Performing a paper chromatography experiment is basically a three-step process: 1) application/treating of the sample 2) "developing" the chromatogram by allowing the mobile phase to move up the paper, and 3) calculating Rf values

R Value
The retention factor (R) may be defined as the ratio of the distance traveled by the substance to the distance traveled by the solvent.

Rf =

distance traveled by substance from application point distance traveled by solvent from application point

It represents the movement or migration of solute relative to the solvent

PIGMENT VISIBLE
Carotene Yellow Alpha Carotene Yellow Orange Xanthopyll Yellow

RF VALUE
0.98 0.97 0.86

Beta Carotene Yellow Orange

Xanthophyll Red

0.94
0.8

Lycopene Yellow Orange Red

Phaeophytin Dark Gray

0.81
0.67

Leutein Yellow Brown

0.75

Phaeophytin Light Gray

Violaxathin Yellow Brown Xanthophyll Yellow Neoxathin Yellow Brown Chlorophyll Light Blue Green Chlorophyll Dark Blue Green Chlorophyll Light Yellow Green Chlorophyll Dark Yellow Green

0.6
0.66 0.5 0.28 0.48 0.46 0.30 0.25

USES OF PAPER CHROMATOGRAPHY

Pathology and Forensic Science
Paper chromatography is useful in the field of forensic science, for investigation of crime. This is because this process can be successfully carried out even with very small quantities of material. Samples from crime scenes are collected to be analyzed and identified, using this technique. Used in DNA and RNA fingerprinting. Pathological laboratories use paper chromatography to detect the presence of alcohol or chemicals in blood.

Qualitative Analysis
Paper chromatography is one of the methods of qualitative analysis, to analyze or separate the different constituents of a mixture. It is a useful tool for separating polar as well as nonpolar solutes. Pharmaceutical companies use this technique to analyze the different compounds in drugs. Used in the testing of antibiotics and determining the pollutants in water.

GAS ABSORPTION PRESSURE DROP MEASUREMENT

PACKED TOWER/COLUMN
Packed towers are vertical columns filled with suitable packing and normally operate countercurrently. Liquid enters the top of the column and is distributed over the top of the column packing via nozzles or distributor plates. Liquid flows downward while contacting with the vapor phase.

 The liquid flows downward through the packing against the pressure and the flowing gas phase because the liquid is appreciably denser than the gas and so is pulled down by gravity. If we keep the flow rate of either liquid or gas constant and increase the flow rate of the other phase, we will eventually come to a limiting condition in which counter-current flow cannot be maintained. This limiting condition is called FLOODING.

 Internal packing provides a large surface area for twophase contact and facilitates transfer of materials between phases.

You can also consult p. 570 of McCabe and Smith 7th edition for more information

ANALYSIS OF PRESSURE DROP IN PACKED COLUMNS

As the flow rate of liquid or gas is increased through a packed column of constant diameter, the pressure drop per foot of packing increases. The pressure drop is greater than that in dry packing, because the liquid in the tower reduces the space available for gas flow.

 When the packing is dry, the line obtained is straight and has a slope of about 1.8. The pressure drop therefore increases with the 1.8 power of the velocity. If the packing is irrigated with a constant flow of liquid, the relationship between pressure drop and gas flow rate initially follows a line parallel to that of the dry column.

 With a dry packing (i.e. no liquid flow, L = 0), pressure drop increases as gas velocity increases according to the linear relationship as shown by line a-a With liquid flowing in the column, the packings now become wetted (irrigated). Part of void volume in the packings now filled with liquid, thereby reducing the cross-sectional area available for gas flow.

 For a constant liquid flow (say L = 5000), at low to moderate gas velocity G; the pressure drop characteristics is similar to that of dry packings, i.e. section b-c of the plot is still straight on log-log plot. Up to this point, there is an orderly trickling of the liquid down the packings. There is no observable liquid being trapped among the packings (no liquid hold-up).

 As the gas velocity is increased further, the pressure drop increased. Some liquid started to be retained in the packings. When point c is reached, the quantity of liquid retained in the packed bed increases significantly. There is a change in slope of the line at point c as pressure drop increases more rapidly with G.Point c is known as the loading point, as liquid starts to accumulate (load) in the packings.

 From point c to d to e, there is a sharp increase in pressure drop at higher G: there is a greater amount of liquid hold-up, a gradual filling of the packing voids with liquid (starting at the bottom of the column), and the column is slowly "drowned" in the liquid.

 At point e, there is another sharp change in the slope. At this point the entire column is filled liquid and the gas now has to bubble through the liquid in the packing voids. The gas pressure drop is now very high. Point e is known as the flooding point. The gas velocity at this point is known as the flooding velocity (limiting velocity)

Points to Note At constant liquid rate, gas pressure drop increases with gas velocity. t constant gas velocity, the gas pressure drop is higher at larger liquid rate. Each liquid rate has its own loading and flooding points. At higher liquid rate, the loading and flooding points occur at lower gas pressure drop. Operation of a gas absorption column is not practical above the loading point. For optimum design, the recommended gas velocity is 1/2 of the flooding velocity.