Evaluation of ODS-AQ stationary phase for use in capillary electrochromatography

The aim of this study was to evaluate the applicability of ODS-AQ packing material as a stationary phase in capillary electrochromatography (CEC). The electroosmotic flow created on an ODS-AQ stationary phase was measured at different mobile phase compositions and at different column temperatures. It was observed that the electroosmotic flow generated in the column increased by 50% when the temperature of the system was raised from 20 degrees C to 60 degrees C, while all other conditions were kept constant. The electroosmotic flow produced by the ODS-AQ stationary phase was found to be comparable to the flow generated in a column packed with Nucleosil bare-silica material. In addition, a set of polar compounds (D-lysergic acid diethylamide derivatives) was utilized to determine the influence of temperature and mobile phase composition on their chromatographic behavior on an ODS-AQ stationary phase in a CEC mode. A linear relationship between the solute retention factor and column temperatures was seen over the temperature range studied (20 degrees C to 60 degrees C). A quadratic function was used to describe the changes in the solute retention factors with variation of acetonitrile concentration in the mobile phase.     

Characterisation of stationary phases in subcritical fluid chromatography with the solvation parameter model. III. Polar stationary phases

In this third paper, varied types of polar stationary phases, namely silica gel (SI), cyano (CN)- and amino-propyl (NH2)-bonded silica, propanediol-bonded silica (DIOL), poly(ethylene glycol) (PEG) and poly(vinyl alcohol) (PVA), were investigated in subcritical fluid mobile phase. This study was performed to provide a greater knowledge of the properties of these phases in SFC, and to allow a more rapid and efficient choice of polar stationary phase in regard of the chemical nature of the solutes to be separated. The effect of the nature of the stationary phase on interactions between solute and stationary phases and between solute and carbon dioxide-modifier mobile phases was studied by the use of a linear solvation energy relationship (LSER), the solvation parameter model. The retention behaviour observed with sub/supercritical fluid with carbon dioxide-methanol is close to the one reported in normal-phase liquid chromatography with hexane. The hydrogen bond acidity and basicity, and the polarity/polarizability favour the solute retention when the molar volume of the solute reduces it. As with non-polar phases, the absence of water in the subcritical fluid allows the solute/stationary phase interactions to play a greater part in the retention behaviour. As expected, the DIOL phase and the bare silica display a similar behaviour towards acidic and basic solutes, when interactions with basic compounds are lower with the NH2 phase. On the CN phase, all interactions (hydrogen bonding, dipole-dipole and charge transfer) have a nearly equivalent weight on the retention. The polymeric phases, PEG and PVA, provide the most accurate models, possibly due to their better surface homogeneity.     

Effects of modifiers in subcritical fluid chromatography on retention with porous graphitic carbon

The effect of different modifiers in subcritical fluid chromatography (SubFC) on interactions between solute and porous graphitic carbon (PGC) and between solute and carbon dioxide-modifier mobile phases was studied by the use of linear solvation energy relationships (LSERs). This study was performed to allow efficient optimization of the composition of the carbon dioxide-modifier mobile phase in regard of the chemical nature of the solutes to be separated. With all modifiers tested (methanol, ethanol, n-propanol, isopropanol, acetonitrile, tetrahydrofuran and hexane), the solute/stationary phase interactions are greater than the solute/mobile phase ones. Dispersion interactions and charge transfer between electron donor solute and electron acceptor PGC mainly explain the retention on this surface, whatever the modifier. These interactions are quite constant over the range of modifier percentage studied (5-40%). For acidic compounds, the retention variation is mainly related to the change in the basic character of mobile and stationary phase due to the variation of modifier percentage. Changes in eluting strength are mostly related to adsorption of mobile phase onto the PGC with methanol and acetonitrile, and to the increase of dispersion interactions between the solute and the mobile phase for other modifiers. Relationships between varied selectivities and solvation parameter values have been studied and are discussed in this paper.     

Preparation and Characterization of Sol-Gel SE-30-Coated Silica Stationary Phase for Capillary Liquid Chromatography

A sol-gel chemistry-based polymer coating approach was developed for the preparation of a novel polysiloxane-coated silica stationary phase for capillary liquid chromatography. SE-30, a commercial polysiloxane stationary phase used in gas chromatography, was incorporated into the properly designed sol solution. Then the sol-gel mixture was introduced into a silica gel-packed capillary column by pressure. A thin film of sol-gel SE-30-coating is chemically bonded to the surface of silica gel particles by hydrolytic polycondensation under mild conditions without any free radical cross-linking procedures, therefore the sol-gel approach offers a simple and effective pathway to create a hybrid polymer-coated silica stationary phase. Various factors affecting column making were optimized and discussed in this report. The resulting stationary phase showed good permeability, mechanical robustness, high durability to alkaline mobile phase and satisfactory chromatographic performance in separations of polar and non-polar aromatic compounds. Linear solvation energy relationships (LSERs) studies indicate that the stationary phase has a reversed-phased character with SE-30 providing chromatographic functionality. The solute size and the solute hydrogen bond ability are major factors that principally govern the retention of test solutes.     

Overview of the retention in subcritical fluid chromatography with varied polarity stationary phases

Retention and separation of achiral compounds in supercritical fluid chromatography (SFC) depend on numerous parameters: some of these parameters are identical to those encountered in HPLC, such as the mobile phase polarity, while others are specific to SFC, as the density changes of the fluid, due to temperature and/or pressure variations. Additional effects are also related to the fluid compressibility, leading to unusual retention changes in SFC, for instance when flow rate or column length is varied. These additional effects can be minimised by working at lower temperatures in the subcritical domain, simplifying the understanding of retention behaviours. In these subcritical conditions, varied modifiers can be mixed to carbon dioxide, from hexane to methanol, allowing tuning the mobile phase polarity. With nonpolar modifiers, polar stationary phases are classically used. These chromatographic conditions are close to the ones of normal-phase LC. The addition of polar modifiers such as methanol or ACN increases the mobile phase polarity, allowing working with less polar stationary phases. In this case, despite the absence of water, retention behaviours generally follow the rules of RP LC. Moreover, because identical mobile phases can be used with all stationary phase types, from polar silica to nonpolar C18-bonded silica, the classical domains, RP and normal-phase, are easily brought together in SFC. A unified classification method based on the solvation parameter model is proposed to compare the stationary phase properties used with the same subcritical mobile phase.     

Insights into the Retention Mechanism of Small Neutral Compounds on Octylsiloxane-Bonded and Diisobutyloctadecylsiloxane-Bonded Silica Stationary Phases in Reversed-Phase Liquid Chromatography

The system constants of the solvation parameter model are used to prepare system maps for the retention of small neutral compounds on an octylsiloxane-bonded (Kinetex C8) and diisobutyloctadecylsiloxane-bonded (Kinetex XB-C18) superficially porous silica stationary phases for aqueous mobile phases containing 10–70% (v/v) methanol or acetonitrile. Electrostatic interactions (cation-exchange) are important for the retention of weak bases with acetonitrile–water but not for methanol–water mobile phases. Compared with an octadecylsiloxane-bonded silica stationary phase (Kinetex C18) retention is reduced due to a less favorable phase ratio for both the octylsiloxane-bonded and diisobutyloctadecylsiloxane-bonded silica stationary phases while selectivity differences are small and solvent dependent. Selectivity differences for neutral compounds are larger for methanol–water but significantly suppressed for acetonitrile–water mobile phases. The selectivity differences arise from small changes in all system constants with solute size and hydrogen-bond basicity being the most important due to their dominant contribution to the retention mechanism. Exchanging the octadecylsiloxane-bonded silica column for either the octylsiloxane-bonded or diisobutyloctadecylsiloxane-bonded silica column affords little scope for extending the selectivity space and is restricted to fine tuning of separations, and in some cases, to obtain faster separations due to a more favorable phase ratio. For weak bases larger differences in relative retention are expected with acetonitrile–water mobile phases on account of the additional cation exchange interactions possible that are absent for the octadecylsiloxane-bonded silica stationary phase.     

Characterisation of stationary phases in subcritical fluid chromatography with the solvation parameter model IV. Aromatic stationary phases

The purpose of the present work was to systematically study the chromatographic behaviour of different aromatic stationary phases in a subcritical fluid mobile phase. We attempted to assess the chemical origin of the differences in retention characteristics between the different columns. Various types of aromatic stationary phases, all commercially available, were investigated. The effect of the nature of the aromatic bonding on interactions between solute and stationary phases and between solute and carbon dioxide-methanol mobile phase was studied by the use of a linear solvation energy relationship (LSER): the solvation parameter model. This study was performed to provide a greater knowledge of the properties of these phases in subcritical fluid chromatography, and to allow a more rapid and efficient choice of aromatic stationary phase in regard of the chemical nature of the solutes to be separated. Charge transfer interactions naturally contribute to the retention on all these stationary phases but are completed by various other types of interactions, depending on the nature of the aromatic group. The solvation vectors were used to compare the different phase properties. In particular, the similarities in the chromatographic behaviour of porous graphitic carbon (PGC), polystyrene-divinylbenzene (PS-DVB) and aromatic-bonded silica stationary phases are evidenced.     

Dual‐mode hydrophilic interaction normal phase and reversed phase liquid chromatography of polar compounds on a single column

Adopting a stationary phase convention circumvents problematic definition of the boundary between the stationary and the mobile phase in the liquid chromatography, resulting in thermodynamically consistent and reproducible chromatographic data. Three stationary phase definition conventions provide different retention data, but equal selectivity: (i) the complete solid phase moiety; (ii) the solid porous part carrying the active interaction centers; (iii) the volume of the inner column pores. The selective uptake of water from the bulk aqueous‐organic mobile phase significantly affects the volume and the properties of polar stationary phases. Some polar stationary phases provide dual‐mode retention mechanism in aqueous‐organic mobile phases, reversed‐phase in the water‐rich range, and normal‐phase at high concentrations of the organic solvent in water. The linear solvation energy relationship model characterizes the structural contributions of the non‐selective and selective polar interactions both in the water‐rich and organic solvent‐rich mobile phases. The inner‐pore convention provides a single hold‐up volume value for the retention prediction on the dual‐mode columns over the full mobile phase range. Using the dual‐mode monolithic polymethacrylate zwitterionic micro‐columns alternatively in each mode in the first dimension of two‐dimensional liquid chromatography, in combination with a short reversed‐phase column in the second dimension, provides enhanced sample information.     

Factors influencing retention and resolution of substituted alkylbenzenes in reversed-phase liquid chromatography

Summary The retention data of isomeric xylenes, ethyltoluenes and diethylbenzenes, and of mesitylene, benzene, toluene and ethylbenzene were obtained on a reversedphase column using methanol-water and ethanol-water mobile phases at four different temperatures. This database was used to relate the dependence of solute retention and resolution on the polarity of the mobile phase, solute dipole moment, and column temperature. The additivity of the free energy of the transfer of solute molecules or solute segments from the stationary phase to the mobile phase, was examined for the isomeric compounds. For this, the logarithm of the net retention volume was substituted for the free energy. Deviations from the additivity of free energies indicate that the separation of isomeric substituted alkylbenzenes is governed by their differential interactions with both the polar mobile phase and nonpolar stationary phase. Among the disubstituted alkylbenzenes,ortho-isomers favor the mobile phase more andpara-isomers tend to prefer the stationary phase more. Themeta-isomers are found to follow the additivity rule closely. These trends are amplified as the polarity of the mobile phase is increased indicating that these isomers are resolved better in water-rich mobile phases. These findings are substantiated by analogous results from gas-liquid chromatographic retention data, estimation of dipole moment effects, and examination of the entropic and enthalpic contributions to the net retention volume.Dedicated to Professor Leslie S. Ettre on the occasion of his 70th birthday.     

Controlling the retention in capillary LC with solvents, temperature, and electric fields

Once a suitable stationary phase and column dimensions have been selected, the retention in liquid chromatography (LC) is traditionally adjusted by controlling the mobile phase composition. Solvent gradients enable achievement of good separation selectivity while decreasing the separation time as compared to isocratic elution. Capillary columns allow use of other programming parameters, i.e. temperature and applied electric fields, in addition to solvent gradient elution. This paper presents a review of programmed separation techniques in miniaturized LC, including retention modeling and method transfer from the conventional to micro- and capillary scales. The impact of miniaturized instrumentation on retention and the limitations of capillary LC are discussed. Special attention is focused on the gradient dwell volume effects, which are more important in micro-LC techniques than in conventional analytical LC and may cause significant increase in the time of analysis, unless special instrumentation and (or) pre-column flow-splitting is used. The influence of temperature upon retention is also discussed, and applications where the temperature has been actively used for retention control in capillary LC are included together with the instrumentation utilized. Finally the possibilities of additional selectivity control by applying an electric field over a packed capillary LC column are discussed.     

Effect of tetraalkylammonium salt on the retention of metal chelates in reversed phase-high performance liquid chromatography

Summary The retention of metal chelates with azo dye chelating agents was greatly reduced when tetraalkylammonium salts (TAAS) were added to the aqueous organic mobile phase in reversed phase-high performance liquid chromatography (RP-HPLC). TAAS was very effective for decreasing the retention time of the chelates including positively charged chelates strongly retained on the reversed phase stationary support. This was probably due to the preferential adsorption of TAAS on the support by the ion exchange process and the hydrophobic interaction. The use of TAAS was a promising means for improving column selectivity in RP-HPLC without depending upon the properties of a stationary support and of a mobile phase.     

Using the liquid nature of the stationary phase in countercurrent chromatography. IV. The cocurrent CCC method

The retention volumes of solutes in countercurrent chromatography (CCC) are directly proportional to their distribution coefficients, K(D) in the biphasic liquid system used as mobile and stationary phase in the CCC column. The cocurrent CCC method consists in putting the liquid "stationary" phase in slow motion in the same direction as the mobile phase. A mixture of five steroid compounds of widely differing polarities was used as a test mixture to evaluate the capabilities of the method with the biphasic liquid system made of water/methanol/ethyl acetate/heptane 6/5/6/5 (v/v) and a 53 mL CCC column of the coil planet centrifuge type. It is shown that the chromatographic resolution obtained in cocurrent CCC is very good because the solute band broadening is minimized as long as the solute is located inside the "stationary" phase. Pushing the method at its limits, it is demonstrated that the five steroids can still be (partly) separated when the flow rate of the two liquid phases is the same (2 mL/min). This is due to the higher volume of upper phase (72% of the column volume) contained inside the CCC column producing a lower linear speed compared to the aqueous lower phase linear speed. The capabilities of the cocurrent CCC method compare well with those of the gradient elution method in HPLC. Continuous detection is a problem due to the fact that two immiscible liquid phases elute from the column. It was partly solved using an evaporative light scattering detector.     

Insights into the Retention Mechanism on a Pentafluorophenylpropylsiloxane-Bonded Silica Stationary Phase (Discovery HS F5) in RP-LC

The solvation parameter model is used to elucidate the retention mechanism of neutral compounds on the pentafluorophenylpropylsiloxane-bonded silica stationary phase (Discovery HS F5) with methanol-water and acetonitrile-water mobile phases containing from 10 to 70% (v/v) organic solvent. The dominant factors that increase retention are solute size and electron lone pair interactions while polar interactions reduce retention. A comparison of the retention mechanism with an octadecylsiloxane-bonded silica stationary phase based on the same silica substrate and with a similar bonding density (Discovery HS C18) provides additional insights into selectivity differences for the two types of stationary phase. The methanol-water solvated pentafluorophenylpropylsiloxane-bonded silica stationary phase is more cohesive and/or has weaker dispersion interactions and is more dipolar/polarizable than the octadecylsiloxane-bonded silica stationary phase. Differences in hydrogen-bonding interactions contribute little to relative retention differences. For mobile phases containing more than 30% (v/v) acetonitrile selectivity differences for the pentafluorophenylpropylsiloxane-bonded and octadecylsiloxane-bonded silica stationary phases are no more than modest with differences in hydrogen-bond acidity of greater importance than observed for methanol-water. Below 30% (v/v) acetonitrile selectivity differences are more marked owing to incomplete wetting of the octadecylsiloxane-bonded silica stationary phase at low volume fractions of acetonitrile that are not apparent for the pentafluorophenylpropylsiloxane-bonded silica stationary phase. Steric repulsion affects a wider range of compounds on the octadecylsiloxane-bonded than pentafluorophenylpropylsiloxane-bonded silica stationary phase with methanol mobile phases resulting in additional selectivity differences than predicted by the solvation parameter model. Electrostatic interactions with weak bases were unimportant for methanol-water mobile phase compositions in contrast to acetonitrile-water where ion-exchange behavior is enhanced, especially for the pentafluorophenylpropylsiloxane-bonded silica stationary phase. The above results are compatible with a phenomenological interpretation of stationary phase conformations using the haystack, surface accessibility, and hydro-linked proton conduit models.     

Insights into the retention mechanism on an octadecylsiloxane-bonded silica stationary phase (HyPURITY C18) in reversed-phase liquid chromatography

Plots of the retention factor against mobile phase composition were used to organize a varied group of solutes into three categories according to their retention mechanism on an octadecylsiloxane-bonded silica stationary phase HyPURITY C18 with methanol-water and acetonitrile-water mobile phase compositions containing 10-70% (v/v) organic solvent. The solutes in category 1 could be fit to a general retention model, Eq. (2), and exhibited normal retention behavior for the full composition range. The solutes in category 2 exhibited normal retention behavior at high organic solvent composition with a discontinuity at low organic solvent compositions. The solutes in category 3 exhibited a pronounced step or plateau in the middle region of the retention plots with a retention mechanism similar to category 1 solutes at mobile phase compositions after the discontinuity and a different retention mechanism before the discontinuity. Selecting solutes and appropriate composition ranges from the three categories where a single retention mechanism was operative allowed modeling of the experimental retention factors using the solvation parameter model. These models were then used to predict retention factors for solutes not included in the models. The overwhelming number of residual values [log k (experimental) - log k (model predicted)] were negative and could be explained by contributions from steric repulsion, defined as the inability of the solute to insert itself fully into the stationary phase because of its bulkiness (i.e., volume and/or shape). Steric repulsion is shown to strongly depend on the mobile phase composition and was more significant for mobile phases with a low volume fraction of organic solvent in general and for mobile phases containing methanol rather than acetonitrile. For mobile phases containing less than about 20 % (v/v) organic solvent the mobile phase was unable to completely wet the stationary phase resulting in a significant change in the phase ratio and for acetonitrile (but less so methanol) changes in the solvation environment indicated by a discontinuity in the system maps.     

Electrically driven microseparation methods for pesticides and metabolites. II: on-line and off-line preconcentration of urea herbicides in capillary electrochromatography.

Capillary electrochromatography (CEC) was introduced to the separation of nine important urea herbicides using octadecyl-silica (ODS) capillary columns that were specially designed to allow the realization of a relatively strong electroosmotic flow (EOF) and, in turn, fast separations. The ODS stationary phase was intentionally prepared to have a low surface coverage in octadecyl ligands in order to ensure a strong EOF. This ODS stationary phase of low surface coverage exhibited the usual reversed-phase chromatographic behavior as was manifested by the linearity of plots of log kappa versus the percent organic modifier in the mobile phase. The nature of the organic modifier of the mobile phase influenced the order of elution as well as the separation efficiency of the nine urea herbicides. Mobile phases containing acetonitrile yielded higher separation efficiency (by a factor of 1.5) than methanol-containing mobile phases. This was attributed to the higher mass transfer resistances of the solute in and out of the pores in the presence of the more viscous methanol-containing mobile phases. Due to the relatively strong affinity of the urea herbicides to the ODS stationary phase, on-line preconcentration consisting of prolonged injections allowed the determination of 10(-5) M urea herbicide samples using a UV detector without sacrificing separation efficiency. This was further decreased to 10(-7) M when the prolonged injection was preceded by the injection of a plug of water. The plug of water (the more retentive mobile phase) brought about an enhanced accumulation of the dilute samples into a narrow band at the inlet of the CEC column. When this on-column sample enrichment approach was combined with an off-line sample preconcentration step, which consisted of a solid-phase extraction process, ultra dilute samples of 10(-10) M (0.1 ppb) could be detected.     

反相液相色谱中溶剂强度规律的研究

 以溶质计量置换保留模型 (SDM R)为依据 ,通过研究 3种正链醇同系物溶剂置换剂对 14种正链醇同系物溶质色谱保留行为的影响 ,发现计量置换参数Z(对应 1mol溶质被吸附时从溶质与固定相接触处释放出的溶剂的总摩尔数 )、lgI(与 1mol溶质对固定相亲和势有关的常数 )和 j(与 1mol溶剂对固定相亲和势有关的常数 )均随着同系物置换剂相对分子质量的增大而减小 ,并呈现出线性变化 ,表明溶剂强度与溶剂分子的大小有关 ,分子愈大 ,溶剂洗脱能力愈强 ,并遵循同系物规律。     

Using the liquid nature of the stationary phase in counter-current chromatography V. The back-extrusion method

The main feature of counter-current chromatography (CCC) is that the stationary phase is a liquid as well as the mobile phase. The retention volumes of solutes are directly proportional to their distribution coefficients K(D) in the biphasic liquid system used in the CCC column. Solutes with high K(D) coefficients are highly retained in the column. The back-extrusion method (BECCC) uses the fact that the liquid stationary phase, that contains the retained solutes, can be easily moved. Switching the column inlet and outlet ports without changing the liquid phase used as the mobile phase causes the rapid collapse of the two immiscible liquid phases inside the column, the previously stationary phase being gathered at the new column outlet. Then this previously stationary liquid phase is extruded outside the CCC column carrying the retained solutes. The back-extrusion method is tested with a standard mixture of five compounds and compared with the recently described elution-extrusion method. It is shown that the chromatographic resolution obtained during the back-extrusion step is good because the solute band broadening is minimized as long as the solute is located inside the "stationary" phase. However, a major drawback of the BECCC method is that all solutes are split between the liquid phases according to their distribution ratios when the CCC column equilibrium is broken. The change of flowing direction should be done after a sufficient amount of mobile phase has flushed the column in the classical mode, eluting solutes with small and medium distribution ratios. Otherwise, a significant portion of the solutes will stay in the mobile phase inside the column and produce a broad peak showing after the stationary phase extrusion.     

Changes in peak width and concentration at the inlet and outlet of a chromatographic column

The original plate model of chromatography is extended to the sorption process occurring at the column inlet and the desorption process at the column exit. At the column inlet it is shown that sufficiently wide feed bands undergo no change in concentration but a fall in band width, i.e., the volume of mobile phase occupied by the solute band is reduced. The reduction factor is (1 + k) where k is the mass distribution ratio (capacity factor). Narrower bands suffer partial reduction in both band width and concentration. On desorption at the outlet, however, the change is always in band width and not concentration. A perfect detector registers the true concentration-time profile of the band in the column if the solute mass fraction in the stationary phase is below 10^?3 at the column outlet. The risks of stripping the stationary phase at high solute concentrations in analytical and preparative or production gas chromatography are compared.