Mobile phase viscosity and velocity dependence on protein retention using nonequilibrium chromatographic techniques

Nonequilibrium chromatography (NEC) is an alternative chromatographic procedure for the separation of macromolecules. The retardation of a protein series is studied using a phosphate buffer as a mobile phase with various concentrations of glycerol fraction (used as a viscosity modifier) at different mobile phase velocities and a C1 column with a very low packing particle diameter as a stationary phase. It is shown that the two factors (viscosity and velocity) of the mobile phase constituted important parameters in the retention mechanism of the proteins in NEC. The retardation velocity domain is divided into two regions. For low velocity regions, the protein retention decreased with a mobile phase velocity increase. This retention is enhanced above a critical value of the mobile phase velocity. The transition between the two well-known NEC methods, slalom chromatography and hydrodynamic chromatography, is clearly visualized for the first time for the protein retention of particular values of the mobile phase velocity.     

A chromatographic approach to analyze dansyl amino acid-HP-beta-CD association using macrocyclic antibiotic as the stationary phase

The retention mechanism for a series of D,L-dansyl amino acids in high-performance liquid chromatography is investigated using a teicoplanin stationary phase and hydroxypropyl-beta-cyclodextrin (HP-beta-CD) as the mobile phase additive (0-16mM). A theoretical treatment is developed to determine the HP-beta-CD influence on the equilibrium between the teicoplanin phase and the aqueous medium, respectively. From the experimental data, the association constants of the D,L-dansyl amino acids-HP-beta-CD inclusion complexes are determined and discussed in relation to the enantiomer structure. A thermodynamic study confirms that both the retention and complexation mechanisms are independent of the dansyl amino acid molecular structure and its absolute carbon configuration.     

Rydberg transitions in neon observed by optogalvanic effect in the 830-870 and 380-420 nm regions

New optogalvanic (OG) Rydberg-Rydberg transitions of neon have been observed in the near-infrared region (830-870 nm), using a commercial Fe-Ne hollow cathode. They involve transitions from the 3d[3/2] J=1 and 3d[7/2] J=3,4 levels to high-lying nf levels. In addition, other OG transitions, observed in the blue range, have been completely assigned to ns, nd, ns′ and nd′ Rydberg series excited from the 3p[1/2] J=1 and 3p[5/2] J=2,3 levels of neon. These transitions and assignments allowed us to extend the range of tunable laser calibration on the two edges of the visible range, where there is a lack of available calibration lines, i.e. the near-infrared and the far-blue range, with a 0.01 nm absolute accuracy.     

Mixing layer resonance under high-speed stream forcing

In the majority of fluid–structure interaction problems, the biggest challenge lies in the fundamental understanding of the flow physics. Forced mixing layers is an important phenomenon found in many cases of flow-induced vibrations and acoustics. The response of a mixing layer to high-speed stream acoustic forcing is investigated with a theoretical and experimental approach. Two different experiments demonstrating the fluid mechanic phenomenon are presented. The first experiment consists of a circular jet impinging on a vibrating plate. The second experiment demonstrates the mixing layer resonance in the context of a fluidelastic instability causing high-amplitude vibrations in gas turbine high-pressure compressor rotor blades. Both the plate and the adjacent blade vibration induce an acoustic feedback that propagates within the jet and blade tip clearance flow, respectively. The resonance was found to occur when the feedback wavelength matched either the jet-to-plate or the inter-blade distance. In both experimental cases, the resonance condition has been simply modeled by the coincidence of a 1D feedback wave, which propagates upstream at reduced velocity by the high-speed flow. The coupling between the jet induced mixing layer and the feedback wave is assumed to naturally occur when one of the wave crests reaches the separation edge. The objective of this study is to improve the understanding of the coupling mechanism between an emanating shear layer and the acoustic forcing originating within a fast flow stream. The study is based on a simplified analytical model in order to enlarge the current understanding of the mixing layer receptivity to the more specific case of its response to high-speed stream forcing. To identify the mixing layer resonant modes, an analytical resonance condition is proposed. It is found that the mixing layer response becomes spatially resonant for specific source locations downstream in the high-speed flow. The study also provides an analytical mean to capture the critical source location periodicity that has been experimentally observed. The resulting theoretical prediction of the resonant source locations is in good agreement with the experimental data. Therefore, it supports the stream forced mixing layer analytical model and the proposed spatial resonance condition. The simple 1D reduced speed feedback wave model, which has been used to identify the experimental resonance conditions, is also in good agreement, and thus validated, with the results of this study.     

Comparison of optimization methods in planar chromatography

Summary The optimisation of chromatographic parameters is studied. The Window Diagram and Overlapping Resolution Map (ORM) methods are compared. The latter is developed to take into account the spot diameter and the average plate height variations with the development distance, in order to improve the reliability of the method.     

Preparative isolation of huperzines A and B from Huperzia serrata by displacement centrifugal partition chromatography

The pH-zone refining centrifugal partition chromatography technique was used to separate the two acetylcholinesterase inhibitors huperzines A and B from a crude alkaloid extract of the club moss Huperzia serrata. Complete co-elution of huperzines A and B was initially observed with the well-known methyl tert-butyl ether-acetonitrile-water (4:1:5, v/v/v) solvent system with triethylamine (8mM) as the displacer and methane sulfonic acid (6mM) as the retainer. An efficient biphasic system was designed on the basis of solvent association that provided selectivity in the elution mode: n-heptane/ethyl acetate/n-propanol/water (5:15:35:45, v/v/v/v). Lowering the bridge solvent content (n-propanol) of this system increased the polarity difference between the two phases thus adapting it to the pH-zone refining mode. Thus, the purification of these compounds was achieved using the biphasic system n-heptane/ethyl acetate/n-propanol/water (10:30:15:45, v/v/v/v) with triethylamine (8mM) as the displacer and methane sulfonic acid (6mM) as the retainer.     

Initiation and propagation steps in the equibinary polymerization of butadiene by bis(h3-allylnickel trifluoroacetate)

Polymerization of butadiene by bis(h³-allylnickel trifluoroacetate) in benzene and o-dichlorobenzene solvents yields an equibinary 1,4-polybutadiene, containing equal amounts of cis and trans isomers. Initiation proceeds by addition of the allylic moiety of the initiator to a butadiene molecule. The rate of initiation is high enough to ensure complete consumption of the catalyst for a monomer/catalyst molar ratio of about 10 at 5°C. The propagation exhibits the characteristics of a “living” polymerization: the molecular weight is proportional to the conversion, and at the end of the reaction, the average degree of polymerization is equal to the monomer/catalyst molar ratio. Living polybutadienyl-nickel trifluoroacetate is able to reinitiate not only butadiene polymerization but also allene polymerization. However, for high [monomer]/[catalyst] ratios, conversion-dependent transfer reactions limit the molecular weight to 7000 in benzene and to 70,000 in bulk polymerization in the presence of small amounts of o-dichlorobenzene.     

Role of the Na+ ion on phenol derivatives/hydroxypropyl-beta-cyclodextrin complex formation on porous graphitic carbon phase

The reversed-phase liquid chromatography retention of phenol derivatives was investigated over a concentration range of sodium chloride (0-10(-2) M) and hydroxypropyl-beta-cyclodextrin (HP-beta-CD) (0-35x10(-3) M) using a porous graphitic carbon (PGC) stationary phase and a methanol/water mixture (50:50 (v/v)) as the mobile phase. A theoretical treatment was developed to investigate the effect of the sodium chloride and hydroxypropyl-beta-cyclodextrin on the equilibrium between the solutes with the PGC surface and the aqueous medium, respectively. The thermodynamic parameter variations were calculated using van't Hoff plots. It was expected that the sodium ion acted on the solute-PGC association process by modifying the surface tension of both the bulk solvent and the PGC surface. The phenol derivative/HP-beta-cyclodextrin complexation was shown to be entropically controlled for all the solutes except for the one which contained the -NO2 group in its structure, i.e. the nitro phenol derivative. A comparison of the compensation temperature of the solute-PGC association process when sodium chloride and HP-beta-CD concentration changed in the mobile phase led to the conclusion that these two modifiers acted via a variation in the hydrophobic effect.