Reverse phase ionic compounds

This is the so-called silanophilic activity of the reversed phase. The compounds affine to silanol-groups are usually bases that interact with the acidic silanol groups via ionic interactions. These interactions are energetically much stronger than the usual hydrophobic interactions of reversed phases, which usually lead to wider elution bands for basic analytes on classical reversed phases. 

In general, the compounds best separated by LSC are those which are soluble in organic solvents and are non-ionic. Water soluble non-ionic compounds are better separated using either reverse-phase or bonded-phase chromatography. 

Separation of ionic and nonionic compounds of alkyl ether carboxylates can be done by reverse phase ion pair chromatography [241]. 

Ionic or partially ionic compounds can be chromatographed on reversed-phase columns through the use of ion-pairing reagents. These reagents are typically long-chain alkyl anions or cations that, in dilute concentrations, can increase the retention of analyte ions. For cationic compounds, C5 to CIO alkyl sulfonates are commonly used combinations may also be used... 

In matrix solid-phase dispersion (MSPD) the sample is mixed with a suitable powdered solid-phase until a homogeneous dry, free flowing powder is obtained with the sample dispersed over the entire material. A wide variety of solid-phase materials can be used, but for the non-ionic surfactants usually a reversed-phase C18 type of sorbent is applied. The mixture is subsequently (usually dry) packed into a glass column. Next, the analytes of interest are eluted with a suitable solvent or solvent mixture. The competition between reversed-phase hydrophobic chains in the dispersed solid-phase and the solvents results in separation of lipids from analytes. Separation of analytes and interfering substances can also be achieved if polarity differences are present. The MSPD technique has been proven to be successful for a variety of matrices and a wide range of compounds [43], thanks to its sequential extraction matrices analysed include fish tissues [44,45] as well as other diverse materials [46,47]. 

Reverse-phase chromatography is used mainly for the separation of nonionic substances because ionic, and hence strongly polar, compounds show very little affinity for the non-polar stationary phase. However, ionization of weak acids (or weak bases) may be suppressed in solvents with low (or high) pH values. The effect of such a reduction in the ionization is to make the compound more soluble in the non-polar stationary phase but the pH of the solvent must not exceed the permitted range for bonded phases, i.e. pH 2-8. 

From the theoretical viewpoint, acetonitrile is the most suitable solvent to study the correlation of retention times and log P values of analytes, since the dipole moment (2.44) is nearly equal to that of water (2.55) (Figure 4.4). The electron donor effect can therefore be eliminated, and the elution order is not changed on modification of the acetonitrile-water mixture ratio. The first choice of an eluent should therefore be an acetonitrile-water mixture for non-ionic compounds in reversed-phase liquid chromatography. Methanol, acetone, THF, or DMF can then be added to improve the resolution. 

The selection of the counter-ion and its concentration are important for the separation of ionic compounds in reversed-phase and ion-exchange liquid chromatography. The addition of hydrophobic ions is an especially powerful method and several surfactants can be used as hydrophobic counter-ions. The theoretical column efficiency of ion-pair liquid chromatography is much better than that of an ion-exchange column, and the regeneration of a column is much faster. Thus, if we can control ion-pair liquid chromatography, we can solve a separation problem. (The important background sources in this area are listed at the end of the chapter.)... 

Ion-pair liquid chromatography can be applied to compounds separated by ion-exchange liquid chromatography, and mixtures of ionic and non-ionic compounds are easily separated. The latter separation is difficult by ion-exchange liquid chromatography. Anions can be separated by reversed-phase ion-pair liquid chromatography (Figure 4.18). 

As more ions enter the solution, the rate of the reverse change, recrystallisation, increases. Eventually, the rate of recrystallisation becomes equal to the rate of dissolving. As you know, when the forward rate and the backward rate of a process are equal, the system is at equilibrium. Because the reactants and the products are in different phases, the reaction is said to have reached heterogeneous equilibrium. For solubility systems of sparingly soluble ionic compounds, equilibrium exists between the solid ionic compound and its dissociated ions in solution. 

Gritti, F. and Guiochon, G, Effect of the pH, the concentration and the nature of the buffer on the adsorption mechanism of an ionic compound in reversed-phase liquid chromatography, ii. Analytical and overload band profiles on Symmetry-C-18 and Xterra-C-18, J. Chromatogr. A, 1041, 63, 2004. 

The elucidation of the retention mechanism in ion-pair reversed-phase chromatography using alkyl amines or alkyl sulfonates as hetaerons has evoked significant interest not only for the great potential of the method in the separation of ionic compounds but also for theoretical reasons. 

Gradient Separations of Ionic Compounds 5.4.3.1 Reversed-Phase Chromatography... 

Reversed-phase chromatography is often used to separate both neutral and ionic organic compounds. In this section, some important aspects for the understanding of the behavior of ionic compounds in reversed-phase chromatography are discussed. The important concepts introduced here are the electrical double layer and the electrostatic surface potential. It will be shown that they are essential for the understanding of the elution profile of ionic compounds. These concepts are further explored in the next section where theoretical models for ion-pair chromatography are discussed. 

Reversed-phase ion-pair chromatography is primarily used for the separation of mixtures of ionic and ionizable compounds. In this chromatographic mode, a pairing ion is added to the mobile phase in order to modulate the retention of the ionic solutes. The pairing ion is an organic ion such as alkylsulfonate, alkylsulfate, alkylamine, tetraalkylammonium ion, etc. Here, only a very brief description of the main ideas behind the electrostatic model for ion-pair chromatography is presented. For a complete discussion, the reader is referred to Ref. [7,8] and the references therein. 

Ion-pair chromatography is a popular form of chromatography in which analyte ions can be paired and separated as ion-pairs on a column (14-17). Ion-pairing agents are usually ionic compounds that contain a hydrocarbon chain that enhances the hydrophobicity of the analyte, so that the ion pair can be retained on a reversed-phase column. 

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