Ion 2022 Word
Ion 2022 Word
Ion 2022 Word
Seminar
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INTRODUCTION:
The ion exchange materials of mineral origin include numerous silicates, alumino silicates,
aluminium hydroxide, zirconium phosphate etc.
The ion exchange materials of organic origin are the products of chemical treatment of coal
or lignin, or manmade high molecular organic compounds containing the ionogenic groups.
(A) Styrene
(B) divinylbenzene
An anion exchange resin is a polymer containing amine (or quaternary ammonium) groups
as integral parts of the polymer lattice and an equivalent number of anions, such as
chloride, hydroxyl, or sulphate ions.
(D) Polystyrene
The concentration of the participating ions of opposite charge also plays an important role.
The retained ions can be eluted with the aid of a high concentration of cations or anions
without changing the pH.
The total ion exchange capacity is defined as the number of milliequivalents of exchangeable
ions per gram of dry resin. usually expressed as millimoles per gram of exchanger. The
capacities of the weakly acidic and weakly basic ion exchangers are functions of pH, the
former reaching moderately constant values at pH above 9 and the latter at pH below 5.
V ×N
Capacity=
W
V ml of alkai solution
W weight of resin.
Chromatographic parameters:
1. Rate of ion exchange process: The rate of ion exchange process depends on the rate of
diffusion which is the slowest state on ion exchange process
2. Flow rate: Due to differences in rates of exchange and the fact that they may vary
significantly for different kinds of separation, flow rates are to be controlled (0.5-5
ml/min)
3. Mechanical strength: Polystyrene bead would have little mechanical strength and upon
adding functional group such as sulphonic acid to the polymer,solubility is greatly
enhanced. If the polymer is crosslinked by the incorporation of divinyl benzene,
mechanical strength is imparted to the resin.
The technique used in ion exchange chromatography is generally the so-called liquid
chromatogram, which means that the eluate from the column is investigated either in small
fractions or continuously by observing some convenient physicochemical property related to the
concentration of the eluate.
The main parameters are- resin, particle size, eluent, temperature, flowrate, column
dimensions.
1. Resin:
Nature and properties of ion exchange resins -
Cross linking and swelling are important factors, when more cross linking agent is present, the resin
becomes more rigid and swells less (has small pore size). This makes separations of ions of
different sizes more difficult as they cannot pass through the pores present and it becomes selective
to ions of different sizes.
The nature of resin whether cationic or anionic exchanger, which determines strongly its
Govind and Rashmi K. (2022-23), Govt College of Pharmacy, Bangalore 6
ION-EXCHANGE
selectivity. Cationic resin is selective for cations and vice versa.
Resin capacity (number of me-equivalents of replaceable ions per gram of dry resin) is
important.
Size of ions: For similarly charged ions; the exchange selectivity increases with decrease
in the size of hydrated ions;
Li+ < Na+ < NH4+ < K+ < Rb+ < Cs+ < H+
Polarizability: Highly polarizable ions are more selective. Exchange is preferred for
greater polarizable ions, as;
I- < Br- < Cl- < F-
3. Eluent:
The choice of eluent composition is in most separations more critical than the choice of
resin. Ions that carry different charges are more easily separated than ions of the same
charge.
From dilute solutions containing ions of different charges those of higher charge are
preferentially retained by the resin.
4. Temperature:
High temperature increases the diffusion rate and therefore the number of theoretical plates
in a given column hence improved separations were observed when working at elevated
temperature.
Other advantages which may be gained by working at elevated temperature, arc reduced
influence of non-ionic adsorption forces.
Another advantage which is gained at elevated temperature is that the resistance to flow is
decreased.
In this connection it must be stressed that independent of the temperature level it is necessary
that the temperature be constant over the whole cross-section of the resin bed.
Care should therefore be taken that uneven heating effects from bunsen burners,
radiators, or sunlight are avoided.
In certain separations, e.g., in the separation of amino acids according- to Stein and
Moore it has been observed that the separation can be improved when the temperature is
changed during the course of elution.
5. Flowrate
The flow rate has a high influence on resolution. The height of a theoretical plate
increases with an increased flow rate. An increased flow rate means that the ions are
carried at a faster rate down the column giving them less time to diffuse into and through
the resin particles and thus, are prevented from reaching all exchanging groups.
High flow rates will cause a broadening of the elution curves and thus an increased
overlap of the curves for the ions to he separated.
The present trend in difficult separations is to work with extremely fine particles at not too
low flow rates, the solution being forced through the column with a pump and thus giving
a constant feed. In this way much time can be saved.
A high flow rate is necessary in separations of short-lived radioactive products and with
such systems.
In general, flow rates are in the range of 0.5 mL/minute to 5 mL/minute.
Govind and Rashmi K. (2022-23), Govt College of Pharmacy, Bangalore 8
ION-EXCHANGE
Fig: Effect of flow rate on IEX resolution. Separation of a 5 ml sample of myoglobin (peak 1),
ribonuclease A (peak 2), and cytochrome c (peak 3) on a 1 x 13 cm (8.7 ml)
7. Particle size:
Partical size contribute directly to rate of exchange.
Large surface area and small particles will increase the rate.
The particle size that is used will depend on the length of column, diameter and whether gravity
or pumping is used to control the flow of the mobile phase.
Most laboratory applications are successful with 100-200 mesh (0.147-0.074 mm).
Smaller particles provide higher resolution but typically also require lower flow rates.
Larger particles permit higher flow rates but yield lower resolution.
Small-particle resin yields sharp, distinct peaks
8. pH value:
pH value is an important parameter for separation and can be controlled and adjusted
carefully by means of buffer substances.
In order to prevent variation in matrix and protein net charge, maintenance of a constant
mobile phase pH during separation is essential to avoid pH changing which can occur when
both protein and exchanger ions are released into the mobile phase.
By means of buffer substances pH value can be controlled and adjusted.
Concentration of H+ and the buffering component influence the protein binding to the
stationary phase, chromatographic resolution and structural as well as functional integrity of
the protein to be separated.
Thus a suitable pH range, in which the stability of sample is guaranteed, has to be identified.
APPLICATIONS:
Ion exchange chromatography can be applied for the separation and purification of many
charged or ionizable molecules such as proteins, peptides, enzymes, nucleotides, DNA,
antibiotics, vitamins and etc.
Ion exchangers are usually employed in a method for the separation of micro as well as
macro samples and are therefore, of practical use in quantitative analysis and purification.
Because of the macro application, ion exchangers can also be used on a pilot plant and
industrial scale. In general, the basic laboratory techniques common to adsorption and
partition column methods are also applied in ion-exchange column methods.
Govind and Rashmi K. (2022-23), Govt College of Pharmacy, Bangalore 1
ION-EXCHANGE
Some of the important applications are discussed with the following examples:
A wide variety of very complex mixtures have been separated using ion exchangers.
These include closely related elements such as Zr and Elf, rare earths, transuranium
elements, halides, alkali metals, and transition metals. Complex mixtures of organic
acids and bases, amino acids, peptides, and proteins have also been separated.
(a) Separation of lanthanides; During the second world war the separation of fission-
produces presented a major problem to analytical chemistry, To
separate these, synthetic resins were employed in combination with the technique
of chromatographic elution.
present in the ion exchange reaction gets held near the top while the other cations to
undergo exchange reaction.
(c) Separation of Sugars: This method was developed by Khym and Zill (1951). These
sugars are first of all converted into borate complexes. Separations of borate
complexes of sugars have been achieved on 11 x 0.9 cm columns of 200
—400 mesh Dowex 1 resin using a loading of 5-10 mg of borate complex and
flow rates 0.5-1 ml/min. Quantitative recovery of sugars is possible after separation of
the borate complexes. Similarly disaccharides can be separated from the
monosaccharides and the individual compounds of hexose and pentose mixtures
resolved.
Ion exchange is used for water and wastewater treatment, primarily for removal of
"hardness ion" (Ca2+ and Mg2+) from water supplies, and for removal of iron and
from ground water supplies. It is also used for treatment of a variety of industrial
wastes to allow for recovery of valuable wastes materials or by products, particularly
ionic forms of precious metals such as silver, gold, and uranium. An
4) Water-softening
The removal of the hardness ions of Ca2+ and Mg2+ may be achieved with a strong acid
cation exchanger in the Na+ form. At low concentrations of most raw waters the resin
strongly prefers the divalent cation over sodium ion, so that the exchange reaction
proceeds to the right.
2RSO3-Na+ + Ca2+ (RSO3)2Ca2+ + 2Na+
5)De-alkalisation
In a water containing bicarbonates — most waters in Western and Central Europe do — calcium and
magnesium associated with bicarbonate ions are exchanged for hydrogen ions from a weakly acidic
cation exchange resin. This is called removal of temporary hardness.
The salinity of the treated water is lower than that of the feed water.
6)Routine analysis of amino acid mixtures, the 20 principal amino from blood serum or from the
hydrolysis of proteins are separated by ion exchange chromatography.
PRACTICAL APPROACH
Source: Coprinus comatus
Extraction: Stipe powder of C. comatus (100 g) was extracted three times with 1 L 95% ethanol
Eluent: 0.2 M sodium phosphate buffer (pH 6.0), and linear gradient of 0.3–1.5 M NaCl in 0.2 M
sodium phosphate buffer (pH 6.0).
Limitations:
I. Even though ion exchange liquid chromatography is considered one of the most
efficient purification methods available, it has short comings as well. Compared
to other purification methods, separation efficiency is low.
II. Secondly, synthesis of ion exchangers is very complex, which slows down the
development of the technique. .
III. Moreover, polystyrene or other aromatic matrices have denaturing effects, leading
to possible dissociation of protein-protein, protein-RNA, -DNA complexes.
However, other matrices preserve the native structure of proteins well.
IV. Sometimes the swollen matrix by volume change complicates the
chromatographic operation as well. But, these drawbacks are quite minor and the
numerous applications described earlier prove its practical efficiency.
Review articles: