Separation of U and Pu by countercurrent chromatography with support-free liquid stationary phase in the TBP white spirit nitric acid system

The article deals with the use of countercurrent chromatography (CCC), support-free partition chromatography, for separation of U and Pu from the organic extract obtained directly by the dissolution of MOX fuel in supercritical CO₂ containing the complex TBP · nHNO₃. White spirit solutions with various TBP concentrations were used as a stationary phase. The effects of the compositions of the stationary and mobile phases on the U/Pu partition efficiency are studied. The CCC method allows the separation of U and Pu under conditions of a TBP concentration gradient in the stationary phase and also of an HNO₃ concentration gradient in the mobile phase. Chromatographic separation first gives the Pu fraction containing 98.9% of Pu and 0.07% of U, and then the U fraction (99.93% of U and 1.1% of Pu). The separation time is 50 min.     

Gas-liquid chromatography with a volatile "stationary" liquid phase

A unique type of gas-liquid chromatography is described in which both mobile and "stationary" phases are composed of synthetic mixtures of helium and carbon dioxide. At temperatures below the critical point of the binary mixture and pressures above the vapor pressure of pure liquid carbon dioxide, helium and carbon dioxide can form two immiscible phases over extended composition ranges. A binary vapor phase enriched in helium can act as the mobile phase for chromatographic separations, whereas a CO2-rich liquid in equilibrium with the vapor phase, but condensed on the column wall, can act as a pseudostationary phase. Several examples of chromatographic separations obtained in "empty" capillary columns with no ordinary stationary liquid phase illustrate the range of conditions that produce such separations. In addition, several experiments are reported that confirm the proposed two-phase hypothesis. The possible consequences of the observed chromatographic phenomenon in the field of supercritical fluid chromatography with helium headspace carbon dioxide are discussed.     

A study of the enthalpy and entropy contributions of the stationary phase in reversed-phase liquid chromatography

The goal of this study was to elucidate the roles played by the stationary and mobile phases in retention in reversed-phase liquid chromatography (RPLC) in terms of their individual enthalpic and entropic contribution to the Gibbs free energy of retention. The experimental approach involved measuring standard enthalpies of transfer of alkylbenzenes from typical mobile phases used in RPLC (methanol/water and acetonitrile/water mixtures), as well as from n-hexadecane (a simple analogue of the stationary phase) to the gas phase, using high-precision headspace gas chromatography. By combining the measured enthalpies with independently measured free energies of transfer, the entropies of transfer were obtained. This allowed us to examine more fully the contribution that each phase makes to the overall retention. It was found that the standard enthalpy of retention in RPLC (i.e., solute transfer from the mobile phase to the stationary phase) is favorable, due to the large and favorable stationary-phase contribution, which actually overcomes an unfavorable mobile-phase contribution to the enthalpy of retention. Further, the net free energy of retention is favorable due to the favorable enthalpic contribution to retention, which arises from the net interactions in the stationary phase. Entropic contributions to retention are not controlling. Therefore, to a great extent, retention is due to enthalpically dominated lipophilic interaction of nonpolar solutes with the stationary phase and not from solvophobic processes in the mobile phase. Further, our enthalpy data support a "partition-like" mechanism of retention rather than an "adsorption-like" mechanism. These results indicate that the stationary phase plays a very significant role in the overall retention process. Our conclusions are in direct contrast to the solvophobic model that has been used extensively to interpret retention in RPLC.     

Chiral separations performed by enhanced-fluidity liquid chromatography on a macrocyclic antibiotic chiral stationary phase

The use of enhanced-fluidity liquid chromatography (EFLC) for chiral separations was demonstrated on a macrocyclic antibiotic column, Chirobiotic-V. This technique was compared to high performance liquid chromatography (HPLC) and supercritical fluid chromatography (SFC) for the separation of chiral compounds in normal-phase mode. The highest resolution was always observed for EFLC condition. Higher efficiency and shorter retention time were also observed for most separations with portions of CO(2) in the range of 0-50 mol %. Larger amounts of CO(2) caused efficiency to decrease and retention time to be prolonged. For some separations, the temperature was elevated to bring the mobile phase to the supercritical condition. Improved efficiency was obtained in SFC, whereas resolution and selectivity were worse. The use of EFLC in reversed-phase chiral separations was also tested. Enantiomer resolution improved under the EFLC condition. For the tested methanol/H(2)O mixture, fluoroform provided more significant improvements in chromatographic performance than CO(2) when used as a fluidity enhancing liquid. The use of EFLC instead of HPLC also caused a markedly lower pressure drop across the column for commonly used flow rates. The low-pressure drop will allow the use of longer columns or multiple columns to increase the total efficiency of the separation. Since chiral columns are often inefficient, this attribute may be very important for chiral separations.     

Ice chromatography. Characterization of water-ice as a chromatographic stationary phase

Water-ice has been characterized as a stationary phase for liquid chromatography. Solutes having two or more polar groups are retained on this stationary phase with THF/hexane as the mobile phase, suggesting that multipoint interactions are required for measurable solute retention. Chromatographic separation of phenols or crown ethers on water-ice is possible. The ice surface is expected to provide two different adsorption sites coming from the OH and O dangling bonds. Although the solute partition into the quasiliquid layer is also considered, the dependence of the retention times on the THF concentration implies that the interaction of solutes with the water-ice surface rather than the partition into the quasiliquid layer is responsible for solute retention. A retention model suggests that the number of adsorption sites for a crown ether depends on its ring size, whereas two sites are involved for the retention of phenols having two hydroxyl groups. Although hydroxyl groups can act as both a hydrogen bond donor and an acceptor, the interaction with the ice OH sites, which are exposed to the surroundings in comparison with the ice O sites, is more important. However, when an acyclic polyether is added to the mobile phase, its adsorption onto the water ice surface allows the creation of the O sites that phenols can approach without steric hindrance. In the presence of the polyethers adsorbed on the ice surface, the retention of phenols is enhanced, whereas crown ethers become less retained due to the competitive adsorption of the polyethers.     

Measurement of partition coefficients in waterless biphasic liquid systems by countercurrent chromatography

Countercurrent chromatography (CCC) is a chromatographic separation technique that uses a liquid as a stationary phase. Centrifugal forces are used to immobilize the liquid stationary phase when the liquid mobile phase is pushed through it. In CCC, the solutes are separated according to their liquid-liquid partition coefficients. The solutes studied were the alkylbenzene homologues from benzene to hexylbenzene and some polyaromatic hydrocarbons (PAHs) from naphthalene to coronene. Their liquid-liquid partition coefficients were measured in the five waterless biphasic systems formed by heptane, as the apolar liquid phase of the five biphasic systems, and four dipolar aprotic solvents, dimethyl sulfoxide, dimethylformamide, furfural, and N-methylpyrrolidone, and the polar proton-donor solvent methanol. The coefficients were compared to the corresponding capacity factors obtained by classical liquid chromatography on octadecyl-bonded silica. For the five biphasic solvent systems studied, linear relationships were found between the partition coefficients and the sp(3) and sp(2) hybridized carbon atom number for the alkylbenzene and PAH series, respectively. The sp(2) and sp(3) transfer energies were estimated, and their ratio was used to quantify the solvent selectivity toward aromatic extraction.     

Factor analysis and experiment design in high-performance liquid chromatography: IV. Influence of mobile phase modifications on the selectivity of chalcones on an ODS stationary phase

Walczak, B., Chrétien, J.R., Dreux, M., Morin-Allory, L. and Lafosse, M., 1987. Factor analysis and experiment design in high-performance liquid chromatography. IV. Influence of mobile phase modifications on the selectivity of chalcones on an ODS stationary pase. Chemometrics and Intelligent Laboratory Systems, 1: 177–189.Chromatographic data were determined for a series of 45 chalcones (E-s-cis and Z-s-cis), separated in systems with Zorbax ODS and heptane + 0.5% of a modifier (tetrahydrofuran, dioxane, ethanol, propanol, octanol or dimethylformamide). To discuss changes in chalcone selectivity arising from mobile phase modifications data processing is carried out with the help of hierarchical ascending classification and correspondence factor analysis. The same methods of analysis are used to compare chalcone selectivity in systems with stationary phases of different polarity (Zorbax ODS and LiChrospher 100 DIOL), and with the same mobile phases. A better understanding of retention mechanism is obtained.     

Factor analysis and experiment design in high-performance liquid chromatography: III. Influence of mobile phase modifications on the selectivity of chalcones on a diol stationary phase

Walczak, B., Morin-Allory, L., Chrétien, J.R., Lafosse, M. and Dreux, M., 1986. Factor analysis and experiment design in high-performance liquid chromatography. III. Influence of mobile phase modifications on the selectivity of chalcones on a diol stationary phase. Chemometrics and Intelligent Laboratory Systems, 1: 79–90.Chromatographic data for a series of 45 chalcones were determined for eight chromatographic systems with LiChrospher 100 DIOL as the stationary phase and with heptane + 0.5% of a modifier (dichloromethane, tetrahydrofuran, ethanol, propanol, octanol, dimethylsulphoxide or dimethylformamide) as the mobile phase. Analysis of the data was carried out with the help of hierarchical ascending classification (HAC) and correspondence factor analysis (CFA). Such a strategy of data processing allows normal-phase high-performance liquid chromatography data to be treated as a source of information on the electronic and steric properties of compounds. CFA also helps to give a better insight into the chromatographic process.     

On-column derivatization using redox activity of porous graphitic carbon stationary phase: an approach to enhancement of separation selectivity of liquid chromatography

A new on-column derivatization method based on the redox activity of porous graphitic carbon (PGC) packing materials was presented for enhancement of separation selectivity of HPLC. Two PGC packing materials were used as the solid redox agents as well as the stationary phase, and their redox activities were investigated using trans-1,2-diaminocyclohexanetetraacetate (DCTA) complexes of some metal ions as probe compounds. It was found that the redox property of PGC was modified by treating them with a solution containing a reducing agent, sodium sulfite or hydroxylammonium chloride. The original PGC packings oxidized Co(II)-DCTA to Co(III)-DCTA during elution, while the PGC treated with a reducing agent showed reduction activity converting Co(III)-DCTA to Co(II)-DCTA. These two cobalt complexes do not form their individual chromatographic zones but migrate as a single zone of their mixture on the PGC column contrary to the chromatographic behavior on a C18 bonded silica, on which Co(II)-DCTA and Co(III)-DCTA can be separated. Treatment of the PGC column with a reducing agent solution transforms the oxidative activity of the original PGC packing to a reductive one from the upper part of the column, so that the retention time of the cobalt complex can be controlled by changing the volume of the reducing agent solution to be used for treatment of the PGC column. The selective separation and determination of cobalt in a reference manganese nodule sample by the developed method was demonstrated.     

High-throughput liquid chromatography/mass spectrometry method for the determination of the chromatographic hydrophobicity index

A fast gradient reversed-phase liquid chromatography (LC) method, using an acetonitrile gradient was developed to determine the chromatographic hydrophobicity index (CHI), as reported by Valco et al. (Anal. Chem. 1997, 69, 2022-2029).The analytical method provides retention times, based on UV detection at two different wavelengths, which then are converted into CHI values after calibration with a set of test compounds. The CHI of each compound is measured at three different pH values, 2.0, 7.4, and 10.5; so using an 8-min gradient at each pH value one compound can be analyzed in approximately 24 min. The aim of this work is to improve the throughput of the CHI screening using a LC/MS approach, so the application of the LC/MS technique is an extension of the LC/UV approach previously reported by Valco et al. This approach allows contemporary injection of N compounds into the LC/MS system, the retention time of each compound can be then extracted from the selected ion recording chromatograms. The throughput of the existing screening method could be increased by N times, where N is the number of compounds injected, so only three runs are needed to determine the CHI at three different pH values for a set of N compounds. The highest value of N depends on the total number of channels that can be monitored simultaneously; in the present work, 32 channels were used. This LC/MS method has been tested for a number of commercial products analyzed as mixtures, and data obtained were compared with those coming from the classical LC/UV approach. In the same way, the method was tested for a number of compounds associated with two GlaxoWellcome projects in the antibacterial area. Data reported show that the LC/MS method can be successfully applied for analyzing compounds in mixtures and for compounds with poor UW absorption, which cannot be analyzed with the standard LC/UV method.     

Polymer-coated fibrous materials as the stationary phase in packed capillary gas chromatography

Synthetic polymer filaments have been introduced as the support material in packed capillary gas chromatography (GC). The filaments of the heat-resistant polymers, Zylon, Kevlar, Nomex, and Technora, were longitudinally packed into a short fused-silica capillary, followed by the conventional coating process for open-tubular GC columns. The separation of several test mixtures such as n-alkylbenzenes and n-alkanes was carried out with these polymer-coated fiber-packed capillary columns. With the coating by various polymeric materials on the surface of these filaments, the retentivity was significantly improved over the parent fiber-packed column (without polymer coating) as well as a conventional open-tubular capillary of the same length. The results demonstrated a good combination of Zylon as the support and poly(dimethylsiloxane)-based materials as the coating liquid-phase for the successful GC separation of n-alkanes and polycyclic aromatic hydrocarbons (PAHs), while successful applications for other separations such as poly(ethylene glycol) coating for the separation of alcohols were also obtained. From the results it has been suggested that the selectivity of the fiber-packed column could be tuned by selecting different coating materials, indicating the promising possibility for a novel usage of fine fibrous polymers as the support material that can be combined with newly synthesized coating materials specially designed for particular separations. Taking advantage of good thermal stability of the fibers, the column temperature could be elevated to higher than 350 degrees C with the combination of a short metallic capillary.     

Reversed phase liquid chromatography of organic hydrocarbons with water as the mobile phase

Reversed phase high-performance liquid chromatography (RP-HPLC) is demonstrated for hydrophobic analytes such as aromatic hydrocarbons using only water as the mobile phase. Achievement of reasonable capacity factors for these types of compounds without the need for toxic and costly organic modifiers in the mobile phase is accomplished by substantially decreasing the phase volume ratio of stationary phase relative to the mobile phase volume and by increasing the polarity of the stationary phase relative to stationary phase materials commonly used for RP-HPLC. Applying a stationary phase of trifluoropropylsiloxane, which is a common gas chromatographic stationary phase material, to nonporous glass microspheres yields a stationary phase with a phase volume ratio reduced by about 2 orders of magnitude as compared to common liquid chromatographic packing materials. As a result, a separation was obtained for hydrophobic organic analytes such as benzene, toluene, ethylbenzene, and isopropylbenzene using a water mobile phase at ambient temperature. A separation of sodium benzoate, benzaldehyde, benzene, and butyrophenone is shown in less than 3 min using a water mobile phase and UV/visible absorbance detection. Additionally, the separation of the ionic surfactant species octyl sulfate and dodecyl sulfate in water in less than 3 min, using unsuppressed conductivity detection, is achieved with a separation mechanism based on interactions with the hydrophobic portion of the surfactant. A water mobile phase offers many potential advantages over traditional mixed aqueous/organic solvent systems. In addition to saving on the cost and expense of buying and disposing of toxic solvents and waste, there is less exposure of the operator to potentially harmful solvents. Increased consistency in reproducing retention times can be expected, since there will not be any variability in solvent strength due to slight variations in mobile phase composition. A water mobile phase produces an environment that should provide an inherent advantage of increased signal-to-noise ratio for detection. Additionally, excellent predictions of the octanol/water partitioning coefficient and aqueous solubility for hydrophobic analytes are obtained from a single measurement of the capacity factor in the water mobile phase.     

Superimposed noninterfering probes to extend the capabilities of phase Doppler anemometry

We propose using multiple superimposed noninterfering probes (SNIPs) of the same wavelength but different beam angles to extend the capabilities of phase Doppler anemometry. When a particle is moving in a SNIP the Doppler signals that are produced exhibit multiple Doppler frequencies and phase shifts. The resolution of the measurements of particle size (i.e., by fringe spacing and Doppler frequency) increases with beam angle. Then, with the solution proposed, even with only two detectors several measurements of size can be obtained for the same particle with increasing resolution if we consider higher frequencies in the signal. Several optical solutions to produce SNIPs as well as a signal-processing algorithm to treat the multiple-frequency Doppler signals are proposed. Experimental validations of the sizing of spherical and cylindrical particles demonstrate the applicability of this technique for particle measurement. We believe that this new technique can be of great interest when high resolution of size, velocity, and even refractive index is required.     

Open tubular anion exchange chromatography. Controlled layered architecture of stationary phase by successive condensation polymerization

The preparation and performance of a multilayered stationary phase for open tubular anion exchange chromatography in relatively large bore (75 microm diameter) columns are described. The inner surface of a fused-silica capillary tube is coated with up to 25 successive porous polymeric layers formed by condensation polymerization of a primary amine with a diepoxide. Each layer of the anion exchange stationary phase consists of copolymer of methylamine (MA) and 1,4-butanedioldiglycidyl ether (BDDE). The polymer layers are sufficiently porous or permeable; each successive layer of the stationary phase incrementally increases the observed column capacity and chromatographic performance in the open tubular mode. Even though the column inner diameter is far from optimum for open tubular liquid chromatography, we demonstrate the baseline separation of a suite of inorganic anions (F-, Cl-, NO2-, Br-, NO3-) in a 5 m x 75 microm column coated with 25 layers of the anion exchange polymer using 1 mM KOH eluent and suppressed contactless conductometric detection at a flow rate of 1 microL/min (operating pressure of approximately 1 bar) with a plate count of >30,000. Strategies for construction of microsuppressor devices used in open tubular ion chromatography are discussed.     

Polymer monoliths with low hydrophobicity for strong cation-exchange capillary liquid chromatography of peptides and proteins

Two polymer monoliths were designed and synthesized from commercially available monomers with an attempt to decrease hydrophobicity for strong cation-exchange chromatography. One was prepared from the copolymerization of sulfoethyl methacrylate and poly(ethylene glycol) diacrylate, and the other was synthesized from vinylsulfonic acid and poly(ethylene glycol) diacrylate. Both of the monoliths were synthesized inside 75-microm i.d., UV-transparent fused-silica capillaries by photopolymerization. The hydrophobicities of the two monoliths were systematically evaluated using standard synthetic undecapeptides under ion-exchange conditions and propyl paraben under reversed-phase conditions. The poly(sulfoethyl methacrylate) monolith demonstrated similar hydrophobicity as a monolith prepared from copolymerization of 2-acrylamido-2-methyl-1-propanesulfonic acid and poly(ethylene glycol) diacrylate, and 40% acetonitrile was required to suppress any hydrophobic interactions with peptides under ion-exchange conditions. However, with the use of vinylsulfonic acid as the functional monomer, a monolith with very low hydrophobicity was obtained, making it suitable for strong cation-exchange liquid chromatography of both peptides and proteins. It was found that monolith hydrophobicity could be adjusted by selection of monomers that differ in hydrocarbon content and type of vinyl group. Finally, excellent separations of model protein standards and high-density lipoproteins were achieved using the poly(vinylsulfonic acid) monolith. Five subclasses of high-density lipoproteins were resolved using a simple linear NaCl gradient.