Polybutadiene-coated Zirconia Research Articles

Titania-based stationary phase in separation of ondansetron and its related compounds

Improvements in stationary phase stability have been and remain a great task for research of new stationary phases. Metal oxide-based stationary phases appear as one of perspective alternatives to classical silica based stationary phases regarding to their similar effectiveness, different selectivity, different retention mechanism and mainly better chemical and thermal stability. In this study, the retention behaviour of ondansetron and its five pharmacopoeial impurities on TiO 2-based reversed phase was investigated. The influence of buffer type, pH and concentration on retention was studied. Different types and amount of organic solvent in mobile phase were tested. The effect of temperature and flow rate on separation was investigated. The separation conditions were optimized and developed method validated. The retention parameters – retention time ( t R), retention factor ( k′), theoretical plate number ( N), resolution between peaks due to nearby peaks ( R s) and symmetry factor ( A s) have been compared to parameters achieved on polybutadiene-coated zirconia column. The thermodynamic parameters of retention of analysed compounds – enthalpy, entropy and Gibbs free energy – were calculated and compared to those achieved on polybutadiene-coated zirconia column. This work proves similarity of retention behaviour of ondansetron and its five related compounds on zirconia-based and titania-based stationary phases and potential utilisation of polyethylene covered TiO 2-based reversed stationary phase as an alternative to polybutadiene-coated ZrO 2 stationary phase in pharmaceutical analysis of ondansetron.

High Temperature Liquid Chromatography of Tocol-Derivatives on Polybutadiene-Coated Zirconia Stationary Phases

High temperature reversed-phase high performance liquid chromatography is evaluated as a new approach for the separation of vitamin E isomers (α-, γ-, δ-tocopherol, and α-, γ-, δ-tocotrienol) and α-tocopherol acetate. The separations of each analyte were examined by varying the eluent composition and column temperature. Using a polybutadiene-coated zirconia column at temperatures of 80 °C–140 °C, complete separations were achieved within 10–20 min using organic modifier (acetonitrile) in the range of 40–50%. Irregularity of the Van’t Hoff analysis was noted over the temperature studied. The plot tends to deviate from linearity at higher operating temperatures (140 °C and 150 °C) with linear deviation errors of 12.7% and 41.6%, respectively, for the mobile phase examined.

High temperature liquid chromatography of triazole fungicides on polybutadiene-coated zirconia stationary phase

High temperature liquid chromatography using water-rich and superheated water eluent is evaluated as a new approach for the separation of selected triazole fungicides, hexaconazole, tebuconazole, propiconazole, and difenoconazole. Using a polybutadiene-coated zirconia column at temperatures of 100–150°C, clear separations were achieved when 100% purified water was utilized as organic-free eluent. Excellent limits of detection down to pg level were obtained for the separation of the triazole fungicides under optimum conditions. Van’t Hoff plots for the separations were linear suggesting that no changes occurred in the retention mechanism over the temperature range studied.

Comparison of the chromatography of octadecyl silane bonded silica and polybutadiene-coated zirconia phases based on a diverse set of cationic drugs

In this study, we compare the separation of basic drugs on several octadecyl silane bonded silica (ODS) phases and a polybutadiene-coated zirconia (PBD–ZrO 2) phase. The retention characteristics were investigated in detail using a variety of cationic drugs as probe solutes. The ODS phases were selected to cover a relatively wide range in silanol activity and were studied with ammonium phosphate eluents at pH 3.0 and 6.0. Compared to any of the ODS phases, the PBD–ZrO 2 phase showed very significant differences in selectivities towards these drugs. Due to the presence of both reversed-phase and ion-exchange interactions between the stationary phase and the basic analyte on ODS and PBD–ZrO 2, mixed-mode retention takes place to some extent on both types of phases. However, very large differences in the relative contributions from ion-exchange and reversed-phase interactions on the two types of phases led to quite different selectivities. When phosphate is present in the eluent and adsorbs on the surface, the PBD–ZrO 2 phase takes on a high negative charge over a wide pH range due to phosphate adsorption on its surface. On ODS phases, ion-exchange interactions result from the interactions between protonated basic compounds and ionized residual silanol groups. Since the pH of the eluent influences the charge state of the silanol groups, the ion-exchange interactions vary in strength depending on pH. At pH 6.0, the ion-exchange interactions are strong. However, at pH 3.0 the ion-exchange interactions on ODS are significantly smaller because the silanol groups are less dissociated at the lower pH. Thus, not only are the selectivities of the ODS and PBD–ZrO 2 phases different but quite different trends in retention are observed on these two types of phases as the pH of the eluent is varied. More importantly, by using the large set of “real” basic analytes we show the extreme complexity of the chromatographic processes on the reversed stationary phases. Both the test condition and solute property influence the column performance. Therefore, use of only one or two probe solutes is not sufficient for column ranking.

Effect of amine counterion type on the retention of basic compounds on octadecyl silane bonded silica-based and polybutadiene-coated zirconia phases.

In a previous paper, we compared the mixed-mode retention characteristics of cationic solutes on octadecyl silane-bonded silica (ODS) and polybutadiene-coated zirconia (PBD-ZrO2) phases. It is well recognized that both reversed-phase and ion-exchange interactions contribute to the retention of cations on ODS phases. The reversed-phase interaction results from the bonded hydrocarbon chain; the ion-exchange interaction originates in the ionized residual silanol groups. These two types of interactions also exist on the PBD-ZrO2 phase. The polybutadiene contributes to the reversed-phase interaction and the ionized zirconol, but primarily, the adsorbed Lewis base anions, such as phosphate or fluoride, contribute to the ion-exchange interaction. We have shown that on ODS phases, reversed-phase interactions are much more important, whereas the opposite is true of PBD-ZrO2 phases. In this work, we investigate the effect of several amine mobile phase counterions on the retention of cationic solutes on ODS and PBD-ZrO2 phases. The effects of the chain length and the type of amine (1 degree, 2 degrees, 3 degrees) counterion on the retention of basic compounds were studied. In contrast to older studies of type A silica-based phases, the results show that the chain length and type of the amine blocker do not have a large effect on the retention of basic compounds with the newer type B silica-based materials. However, on the PBD-ZrO2 phase, very striking differences in retention were observed with different amine counterions. We show that the molecular geometry of the amine counterion has a significant effect on the retention of basic solutes on the PBD-ZrO2 phase.

The special effect of fluoride on the chromatography of acidic analytes on polybutadiene-coated zirconia

The special effect of fluoride as a Lewis base additive in suppressing the ligand-exchange interactions for acidic analytes on polybutadiene-coated zirconia (PBD-ZrO2) has been investigated. We found that fluoride is more effective than phosphate in improving the separation efficiency for strong acids. The improvement is attributed to that fluoride has a smaller size and a more flexible coordination chemistry towards zirconium centers than phosphate; consequently, fluoride can more effectively improve the kinetics of the ligand-exchange processes for strongly acidic analytes. We demonstrated that using a small amount of fluoride in combination with a larger quantity of phosphate is a practical way to improve the separation efficiency and resolution for acidic analytes. Some example separations are presented.

Mixed-mode reversed-phase and ion-exchange separations of cationic analytes on polybutadiene-coated zirconia

The retention and selectivity of the chromatographic separation of basic (cationic) analytes on a polybutadiene-coated zirconia (PBD–ZrO 2) stationary phase have been studied in greater detail than in previous studies. These separations are strongly influenced by the chemistry of the accessible surface of zirconia. In the presence of buffers which contain hard Lewis bases (e.g., phosphate, fluoride, carboxylic acids) zirconia’s surface becomes negatively charged due to adsorption of the buffer anion at the hard Lewis acid sites. Consequently, under most conditions (e.g., neutral pH), cationic analytes undergo both hydrophobic and cation-exchange interactions. This mixed-mode retention process generally leads to greater retention factors for cations relative to those on silica-based reversed phases despite the lower surface areas of the zirconia phase, but, more importantly, adsorption of hard Lewis bases can be used to control the chromatographic selectivity for cationic analytes on these zirconia-based stationary phases. In contrast to our prior work, here we show that when mixed-mode retention takes place, both retention and selectivity are easily adjusted by changing the type of hard Lewis base buffer anion, the type of buffer counter-ion (e.g., sodium, potassium, ammonium), the pH, and the ionic strength of the eluent as well as the type and amount of organic modifier.

Separation of selected basic pharmaceuticals by reversed-phase and ion-exchange chromatography using thermally tuned tandem columns.

The separation of basic pharmaceuticals is usually performed on C8 or C18 bonded silica supports. Silanolphilic interactions between basic analytes and surface silanol groups often lead to tailed peaks, poor efficiency, and irreproducible retention times. To solve these problems, many new types of silica-, zirconia-, and polymer-based columns, which provide unique selectivities, improved stability at high pH, or both, have been developed for the analysis of basic compounds. The essence of method development for the chromatographic analysis of basic compounds is to choose a system in which the band spacing can be varied dramatically, quickly, and conveniently while minimizing the tailing due to silanolphilic interactions. The thermally tuned tandem column (T3C) approach has been shown to provide an effective way to adjust stationary-phase selectivity for nonionic compounds. In this study, a tandem combination of an octadecylsilane (ODS) and a polybutadiene-coated zirconia (PBD-ZrO2) phase was used to separate nine antihistamines. Selectivity is tuned by independently adjusting the isothermal temperatures of the two columns. We found dramatic differences in the retention factors, elution sequences, and band spacing for the above set of basic drugs on the two types of columns. The T3C model has been used successfully to locate the optimal temperatures based on only four exploratory runs. The nine antihistamines were baseline separated on the tandem column combination even though they could not be separated on the individual phases. The effect of the buffer concentration on retention of the basic antihistamines was also studied. We conclude that cation-exchange interactions predominate on the PBD-ZrO2 phase, while reversed-phase interactions are more important on the ODS phase. Interestingly, an increase in column temperature causes a significant increase in the retention on the ODS column and a decrease of retention on the PBD-ZrO2 column. This can be explained by the change in the analyte's degree of ionization with temperature. The T3C combination of silica- and zirconia-based RPLC columns is demonstrated to be a powerful approach for the separation of this mixture of basic analytes.

A study of the Lewis acid-base interactions of vinylphosphonic acid-modified polybutadiene-coated zirconia.

Polybutadiene-coated zirconia (PBD-ZrO2) is very useful for reversed-phase separations under a wide variety of conditions. Its excellent chemical (pH = 1-13) and thermal (up to 150 degrees C) stability distinguish it from silica-based reversed phases. Just as with silica-based phases, zirconia's surface chemistry significantly influences the chromatography of certain classes of analytes. Zirconia's hard Lewis acid sites can be chromatographically problematic. Analytes such as carboxylic acids strongly interact with these sites on PBD-ZrO2 and do not elute. Addition of phosphate or other strong, hard Lewis bases to the eluent brings about elution, but the resulting peak is often tailed and broad. Typically, cationic solutes are more retained in the presence of phosphate or fluoride due to adsorption of the Lewis base additives and the concomitant development of a negative charge on the surface. This Coulombic interaction can be used to optimize selectivity, but the reversed-phase-cation-exchange retention can produce broad peaks with excessive retention. As an alternative to adding Lewis bases to the eluent, we studied the effect of permanently modifying PBD-ZrO2 by covalently attaching vinylphosphonic acid (VPA) to PBD which was predeposited in the pores of zirconia. We have investigated the chromatography of acids, bases, and small peptides on VPA-modified PBD-ZrO2 (VPA-PBD-ZrO2) and compared it to PBD-ZrO2. VPA-PBD-ZrO2 is a reversed-cation-exchange phase with properties quite different from PBD-ZrO2. The chemical stability of both phases led us to explore how low-pH (1.5-3), ultralow-pH (0), and high-pH (12) eluents effect the retention properties of these mixed-mode phases. Ultralow-pH eluents effectively separate small peptides on both phases. This approach gives lower retention, without sacrificing resolution, and much higher efficiency for small peptides than previously reported.

Polybutadiene-coated zirconia packing materials in solvating gas chromatography using carbon dioxide as mobile phase

In this paper, practical considerations of column efficiency, separation speed, thermal stability, and column polarity of capillary columns packed with polybutadiene-coated zirconia were investigated under solvating gas chromatography (SGC) conditions using carbon dioxide as mobile phase. When compared with results obtained from conventional porous octadecyl obtained from conventional porous octadecyl bonded silica (ODS) particles, PBD-zirconia particles produced greater change in mobile phase linear velocity with pressure than conventional ODS particles under the same conditions. The maximum plate number per second (Nt) obtained with a 30 cm PBD-zirconia column was approximately 1.5 times higher than that obtained with an ODS column at 100 °C. Therefore, the PBD-zirconia phase is more suitable for fast separations than conventional ODS particles in SGC. Maximum plate numbers per meter of 76,900 and 63,300 were obtained using a 57 cm×250 μm i.d. fused silica capillary column packed with 3 μm PBD-zirconia at 50 °C and 100 °C, respectively. The PBD-zirconia phase was stable at temperatures up to 320 °C under SGC conditions using carbon dioxide as mobile phase. Polarizable aromatic compounds and low molecular weight ketones and aldehydes were eluted with symmetrical peaks from a 10 cm column packed with 3 μm PBD-zirconia. Zirconia phases with greater inertness are required for the analysis of more polar compounds by SGC.

Fast separations at elevated temperatures on polybutadiene-coated zirconia reversed-phase material.

This paper describes the results of thermal stability studies of polybutadiene-coated zirconia reversed phases and application to fast HPLC separations at elevated temperatures and high flow rates. The thermal stability of the material was evaluated at temperatures up to 200 degrees C, and the rapid analyses of polyaromatic hydrocarbons and typical reversed-phase test mixtures were carried out at 100 degrees C and a flow rate of 5 mL/min. We found that the material is completely stable at 200 degrees C for at least 1300 column volumes. Analysis time can be decreased about 18-fold at high temperatures and flow rates without any significant loss in resolution relative to that at conventional temperatures and normal flow rates. For the separation of a five-component reversed-phase test mixture, the analysis time was only 50 s.

Evaluation of temperature effects on selectivity in RPLC separations using polybutadiene-coated zirconia.

The effect of temperature on selectivity in RPLC method development has been evaluated on polybutadiene-coated zirconia. We find that the influence of temperature on selectivity depends strongly on solute type. For solutes of similar structure such as polyaromatic hydrocarbons, temperature has almost no effect on selectivity; however, for solutes with very different functional groups such as chlorophenols, temperature changes did significantly affect selectivity. We feel that simple mixtures with one dominant retention mechanism-e.g., solvophobic retention-will not be helped appreciably by adjusting temperature. However, in complex mixtures with polar and ionizable solutes, optimization by varying the temperature may well be fruitful.

A study of the efficiency of polybutadiene-coated zirconia as a reversed-phase chromatographic support.

This work describes the dynamic characteristics of columns packed with polybutadiene (PBD)-coated zirconia. Reduced plate height (h) vs reduced velocity (v) data for alkylbenzenes were acquired for three phases coated with different amounts of PBD. Additional data for benzene (k' = 0) on uncoated and four coated phases were also collected. These h vs v plots have a minimum between 4 and 5, indicating good but not excellent column efficiency. By fitting the Knox equation to these data, the A (packing quality), B (axial diffusion), and C (mass transfer resistance) coefficients were determined. The A coefficients were about 2 in all cases, and the B coefficients had values between 2 and 3. The C coefficients varied from 0.05 to 0.2, depending on the polymer load. The data suggest that the quality of packing and the efficiency of mass transfer resistance in the "film" of polymeric stationary phase, not the stagnant mobile phase, are key to further improvements in column performance. The amount of polymer loaded significantly affects the efficiency through the mass transfer resistance in the stationary phase.

Revisionist look at solvophobic driving forces in reversed-phase liquid chromatography IV. Partitioning vs. adsorption mechanism on various types of polymeric bonded phases

The partition and adsorption mechanisms of retention in reversed-phase liquid chromatography have been examined based on a comparison of the free energy of transfer of methylene groups from aqueous-organic mixtures to bulk hexadecane with those to a variety of polymeric bonded phases. The stationary phases studied include: conventional silica-based polymeric phases of various alkyl chain lengths, a so-called “horizontally polymerized” octadecyl phase on silica and a series of polybutadiene-coated zirconia phases. The data indicate that for methylene groups a partition-like mechanism is dominant on all phases. On the polybutadiene-coated zirconia and “horizontally polymerized” octadecyl phases the partition mechanism holds at all mobile phase compositions. In contrast on conventional polymeric silica phases the retention mechanism seems to become more adsorption-like at methanol compositions greater than about 70% (v/v).

Effect of temperature on the thermodynamic properties, kinetic performance, and stability of polybutadiene-coated zirconia.

This article describes the results of a study of the effect of temperature on the performance of a reversed-phase material prepared by coating polybutadiene (PBD) on porous zirconia. We examined the effect of temperature on retention, efficiency, and stability of this phase. The thermodynamic properties were evaluated via the separation of alkylbenzenes and a set of tricyclic antidepressant drugs at different temperatures, while the intrinsic kinetic performance of the PBD phase at elevated temperatures was examined by using alkylbenzenes as probe solutes. Moreover, the thermal stability was determined by measuring the drift in k' while continuously pumping a mobile phase at 100 degrees C. We found that enthalpy changes were between -2 and -4 kcal/mol and that changes in selectivity varied with the type of solute. High temperatures improved the column efficiency by 30%, mainly by accelerating the solute diffusion rate in the stationary phase. Finally, the PBD-coated zirconia phase was very stable at a temperature of 100 degrees C for at least 7000 column volumes.

Characterization of polybutadiene-coated zirconia and comparison to conventional bonded phases by use of linear solvation energy relationships

This paper describes the use of linear solvation energy relationships (LSERs) to thermodynamically characterize retention on a reversed-phase type material based on polybutadiene (PBD)-coated zirconia. Retention data were obtained for a large set of judiciously selected test solutes on four PBD phases (carbon loads varying from 1.5 to 5.6% by weight) in various methanol-water mobile phases. Based on the LSERs, the free energy of retention was dissected into contributions from cavity formation/dispersive interactions, dipolar interactions, and hydrogen bond (HB) donor-acceptor interactions. The PBD-zirconia phases were compared to conventional silica-based bonded reversed phases. As is the case with conventional bonded phases, the solute's size and HB acceptor basicity are the predominant retention determining factors, and on the whole, PBD-zirconia phases closely resemble conventional chemically bonded reversed phase materials. Interestingly, the solute's basicity has a larger effect on its retention on the PBD-zirconia phase than on conventional bonded phases, so relative to their behavior on conventional phases, strong hydrogen bases and highly dipolar analytes, when compared to nonpolar solutes, are less strongly retained on PBD-zirconia than on conventional phases. PBD-zirconia and conventional phases are so similar that there should be little difficulty in transferring separation methods between phases.

Chromatography of Proteins Using Polybutadiene-Coated Zirconia

Polybutadiene-coated zirconia (PBD-ZrO2), when used as a stationary phase in conjunction with a mobile phase containing phosphates, constitutes a reversed-phase/cation-exchange mixed-mode chromatographic system. The separation of proteins on this phase can be achieved only through the use of mobile phases containing the correct combination of phosphoric acid, displacing salt, and organic cosolvent. We found that excessive Coulombic interactions between proteins and the stationary phase impair the system performance for the separation of proteins. The effects of mobile phase conditions on the separation of proteins using phosphate-adsorbed PBD-ZrO2 are studied in this work. Factors such as the presence of a multivalent cation, mobile phase pH, phosphate concentration, and salt concentration can be manipulated to reduce the net negative charge on the surface and thereby improve the performance of the system toward the separation of proteins.

Mixed-mode retention of peptides on phosphate-modified polybutadiene-coated zirconia.

Zirconia HPLC packing materials were found to be potentially advantageous for large-scale protein separations due to their excellent pH stability and mechanical stability. However, Lewis acid sites on zirconia's surface cause irreversible adsorption of proteins due to their interactions with hard Lewis bases such as the carboxyl groups in proteins. Although the Lewis acid sites can be effectively blocked by adsorbing phosphate ions onto zirconia's surface, proteins and peptides cannot be eluted using a typical reversed-phase mobile phase. In this work, we found that the separation of peptides on a phosphate-modified polybutadiene-coated zirconia (PBD-ZrO2) can be brought about by using a mobile phase containing both an organic modifier and a high concentration of sodium perchlorate. The salt is needed to cancel the Coulombic interactions between the negatively charged stationary phase and the positively charged proteins. To understand the retention mechanism of proteins and peptides on phosphate-modified PBD-ZrO2, this work was aimed at the study of the surface characteristics of the phosphate-modified PBD-ZrO2. We found that the phosphate-modified PBD-ZrO2 phase has both reversed-phase and cation-exchange characteristics under the acidic mobile-phase conditions used for protein and peptide separations. The PBD coating provides hydrophobic moieties, and the phosphate ions adsorbed on zirconia's surface provide cation-exchange sites. Reversed-phase separation of a peptide standard mixture and cation-exchange separation of a cationic peptide standard mixture on the same phosphate-modified PBD-ZrO2 column shows excellent column resolution in both modes. Although mixed-mode stationary phases provide unique selectivity, the secondary equilibrium on phosphate-modified PBD-ZrO2 can cause peak broadening. Applications of the phosphate-modified PBD-ZrO2 to peptide separations are demonstrated here.

Study of the irreversible adsorption of proteins on polybutadiene-coated zirconia

The cause of irreversible adsorption of proteins on polybutadiene-coated zirconia is investigated by comparing the chromatographic properties of polybutadiene-coated zirconia with that of other reversed-phase packing materials such as bonded phase silica, polybutadiene-coated alumina and polybutadiene-coated silica. We find that the polybutadiene-coated zirconia has a micropore size distribution similar to that of the polybutadiene-coated alumina, from which some proteins can be eluted. Thus, the irreversible adsorption of proteins on polybutadiene-coated zirconia is not caused by entrapment of proteins in the micropores of the packing. The high hydrophobicity of the polybutadiene coating and the strong Lewis acid sites on the zirconia surface cause strong interactions between proteins and the stationary phase, the combination of which lead to irreversible adsorption of proteins on polybutadiene-coated zirconia.