Retention surface silanols

Silica has often been modified with silver for argentation chromatography because of the additional selectivity conferred by the interactions between silver and Jt-bonds of unsaturated hydrocarbons. In a recent example, methyl linoleate was separated from methyl linolenate on silver-modified silica in a dioxane-hexane mixture.23 Bonded phases using amino or cyano groups have proved to be of great utility. In a recent application on a 250 x 1-mm Deltabond (Keystone Scientific Belief onte, PA) Cyano cyanopropyl column, carbon dioxide was dissolved under pressure into the hexane mobile phase, serving to reduce the viscosity from 6.2 to 1 MPa and improve efficiency and peak symmetry.24 It was proposed that the carbon dioxide served to suppress the effect of residual surface silanols on retention. 

Jt is not possible to bond all of the surface silanol groups. Unreacted silanols are capable of adsorbing polar molecules, and will thus affect the chromatographic properties of the bonded phase. Usually, the unreacted silanols produce undesirable effects, such as tailing and excessive retention in reverse phase separations, although there have been cases reported where the unreacted silanols improve such... 

Reversed-phase liquid chromatography shape-recognition processes are distinctly limited to describe the enhanced separation of geometric isomers or structurally related compounds that result primarily from the differences between molecular shapes rather than from additional interactions within the stationary-phase and/or silica support. For example, residual silanol activity of the base silica on nonend-capped polymeric Cis phases was found to enhance the separation of the polar carotenoids lutein and zeaxanthin [29]. In contrast, the separations of both the nonpolar carotenoid probes (a- and P-carotene and lycopene) and the SRM 869 column test mixture on endcapped and nonendcapped polymeric Cig phases exhibited no appreciable difference in retention. The nonpolar probes are subject to shape-selective interactions with the alkyl component of the stationary-phase (irrespective of endcapping), whereas the polar carotenoids containing hydroxyl moieties are subject to an additional level of retentive interactions via H-bonding with the surface silanols. Therefore, a direct comparison between the retention behavior of nonpolar and polar carotenoid solutes of similar shape and size that vary by the addition of polar substituents (e.g., dl-trans P-carotene vs. dll-trans P-cryptoxanthin) may not always be appropriate in the context of shape selectivity. 

Table 1 gives the tailing factor for the basic analyte amitriptyline at neutral pH on several commercially available packings. One can clearly see the difference between the older packings and the newer packings based on high-purity silicas. It is unquestionable today that surface silanols on a packing participate in the retention of basic analytes, and... 

The theoretical treatment outlined above does not involve any attractive interaction biptween stationary phase and eluite besides that caused by van der Waals>force. Consequently, the above obscrvniion is not predicted by the ther y. Nevertheless, surface silanols can have profound effect on the selectivity of the stationary phase in contact with eluents rich in organic solvent. The phenomenon and its practical importance in RPC of polar substances, particularly those carrying positive charge, have long been appreciated, Yet, only recently has retention behavior analyzed... 

Factors that influence the retentive powers and selectivity of such bonded phases include the surface concentrations of hydrodartenaceous ligates and free silanol groups. The thermodynamic aspectitm solute interactions with the hydrocarbonaceous ligates at the surface, which are hydrophobic interactions in the case of aqueous eluents, are discussed later in this chapter within the framework of the solvophobic theory. In practice, however, solute interactions with surface silanol which may be termed silanophilic interactions can also contribute ]to retention (71, 75, 93), particularly in the case of amino compounds. Consequently the retention mechanism may be different from that which would be ol served with an ideal nonpolar phase. Therefore, increasing attention is paid to the estimation of the concentration of accessible sianols and to their elimination from the surface of bonded phases. 

If the retention enthalpies of the two sites differ, curvature may be observed in the plots. Moreover, if the enthalpies are opposite in sign, a minimum will occur in the van t Hoff plot at a temperature where the ratio of the retention foctors for the two mechanisms equals the absolute value of the reciprocal of the ratio of the corresponding enthalpies. Most frequently, however, less dramatic curvature would be expected. Such behavior may be anticipated in the RPC of amines with- arge nonpolar moieties which could be retained by silmiophilic interactions with surface silanols and by solvophobic interactions with nonpolar ligates of a reversed phase with low surface coverage. Recently an lihalysis of this behavior has been reported 93). 

Retention in RP chromatography is based on the interaction of the hydrophobic part of the analyte with the hydrophobic section of the stationary phase. This interaction can be modulated with the type and the concentration of the organic modiher in the mobile phase. The selectivity is mainly inflnenced by the interaction of the polar fnnctional gronps of the analyte with constituents of the mobile phase (bnffer, salts, etc. in the aqneons part) and with the amonnt and activity of residual surface silanols, which are, of course, also modihed by mobile phase constituents. 

Each of the types of SPE sorbents discussed retains analytes through a primary mechanism, such as by van der Waals interactions, polar dipole-dipole forces, hydrogen bonding, or electrostatic forces. However, sorbents often exhibit retention by a secondary mechanism as well. Bonded silica ion-exchange sorbents primarily exhibit electrostatic interactions, but the analyte also experiences nonpolar interaction with the bonded ligand. Nonpolar bonded silicas primarily retain analytes by hydrophobic interactions but exhibit a dual-retention mechanism, due to the silica backbone and the presence of unreacted surface silanol groups [72], Recognition that a dual-... 

To understand the role of the surface silanols and their contribution to retention, these compounds were separated on a column that was purposely coated to half of the Cjg level of the original Clg (Fig. 5-21b) (coated column had approximately 50% silanol content). If peak tailing is due solely to the silanol interaction, the peak symmetry should be worse than on the fully coated column. But there was no decrease in peak symmetry. In fact, retention of the tetracaine (peak 4) increased slightly while the peak symmetry improved. Thus, it appears that silanols do contribute to retention but the amount of silanols is not the main cause of peak asymmetry. The logical extension of this approach is to separate the compounds on unbonded silica gel, as shown in Figure 5-27c. In this situation, peak symmetry is quite good and retention is decreased. 

The chromatograms in Figure 5-27 indicate that the surface silanols by themselves are not deleterious to the retention of organic amines when using organic, aqueous eluents containing an inorganic salt. In fact, silica gel itself appears to be the preferred adsorbent for the separation of these bases. The... 

Bij, K.E., Horvath, C., Melander, W.R. and Nahum, A. (1981) Surface silanols in silica-bonded hydrocarbonaceous stationary phases. II. Irregular retention behavior and effect of silanol masking. J. Chromatogr. Z03, 65-84. 

The number and nature of unreacted surface silanols affects the character of a stationary phase. Initially free, geminol or associated silanols are minimized through a process known as endcapping, which bonds various species to the residual silanols. Hydrophilic endcaps or bulky steric endcaps that separate the hydrocarbon chains and prevent analyte interaction with the silica surface can be used. If residual silanols are left unreacted (and some always are), the analyte will be separated based on a combination of interactions with both the reverse-phase support and the highly polar silanol groups. Increased retention, changes in elution order, and tailing will result for basic compounds. 

In RPLC, acidic pH of 2.5-3 is used for many applications. The low pH suppresses the ionization of weakly acidic analytes, leading to higher retention.2,3,11 Surface silanols are not ionized at low pH, lessening tailing with basic solutes. Most silica-based bonded phases are not stable below pH 2 due to acid catalyzed hydrolytic cleavage of the bonded groups.23 Common acids used for mobile phase preparations are phosphoric acid, trifluoroacetic acid (TFA), formic acid, and acetic acid. However, basic analytes are ionized at low pH and might not be retained. 

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