Mobile phase in ion-exchange chromatography

A column used to minimize the conductivity of the mobile phase in ion-exchange chromatography. 

In ion-exchange chromatography (lEC), the mobile phase modulator is typically a salt in aqueous solution, and the stationary phase is an ion-exchanger. For ddnte conditions, the solute retention faclor is commonly found to be a power-law function of the salt uormahty [cf. Eq. (16-27) for ion-exchange equilibrium]. 

Retention of solutes in ion-exchange chromatography is determined by the nature of the sample, the type and concentration of other ions present in the mobile phase, the pH, temperature, and the presence of solvents. Because there are so many variables, it is often not easy to predict what will happen in an ion-exchange separation if we change the experimental conditions. There are some useful guidelines, and to see how they work we will look at the ion-exchange separation of two weak acids (see Fig. 3.3c). 

Modern pump designs also include a means for flushing the piston with solvent behind the pump seal (not shown in Figure 13.4). The solvent for this is drawn in from a separate reservoir and pumped back into this same reservoir. The purpose is to continuously rinse the piston free of mobile phase residue such that abrasive solute crystals resulting from a mobile phase that has dried out on the piston will not deposit there. These solutes, such as the salts dissolved in the buffered mobile phases used in ion exchange chromatography, may otherwise crystallize on the piston and then damage the piston or the pump seal when the piston moves back and forth. Mobile phases that contain such solutes must be flushed from the system after use so that there is also no crystallization on the front side of the seal. 

Analyte retention and selectivity in ion-exchange chromatography are strongly dependent on the pH and ionic strength of the mobile phase. Basic principles of the ion-exchange HPLC are discussed in Chapter 4, Section 4.10. 

Ion-interaction chromatography is an intermediate between reversed-phase and ion-exchange chromatography. Introduction of amphiphilic and Uo-philic ions into the mobile phase causes their adsorption on the hydrophobic surface of packing material with subsequent transformation into a pseudo ion-exchange surface. Ionic interactions with charged analytes can occur in the mobile phase and with counterions that may be adsorbed on the stationary-phase surface. 

Provided that the mobile phase containing counterion C at a concentration (Cl = y) is used in ion-exchange chromatography of trace amounts of ion 5 (analyte), the relationship between the retention factor k of the ion S and the concentration tp can be described by Eq. (1.21) [35] ... 

The retention factor, Eq. (7.2), for each species / is calculated knowing the dead time, t(), and the retention time of species i at infinite dilution, /r,./- There are known methods in the literature for calculating the dead time or retention time for a non-retained peak in normal-phase, reversed-phase and ion-exchange chromatography [67]. For example, in normal-phase chromatography, pentane in 95 5 hexane-acetone is unretained. In reversed-phase chromatography, a common measure of void volume is from the refractive index response obtained when the sample solvent composition is different from the mobile-phase composition. 

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