Sol-gel multicapillary columns for gas-solid chromatography

In this work, we report the method for the preparation of multicapillary columns (MCCs) for gas-solid chromatography. The porous layer adsorbent is formed on capillary walls by the hydrolysis of aluminum alkoxide in the presence of polypropylene glycol (PPG) and HCl. Porosity and selectivity of the adsorbent depend on reaction conditions and the concentration of PPG. Sol-gel MCCs are well suited for high-speed chromatographic analysis of light hydrocarbons by gas-solid chromatography. Nine-component mixtures of C1-C4 hydrocarbons are separated within 8-12 s. The efficiency of 25-30 cm long alumina sol-gel MCCs consisting of approximately 1400 capillaries of 40 microm diameter is up to 2500-3000 theoretical plates.     

Supercritical-fluid chromatography: Open columns vs packed columns

Supercritical-fluid chromatography (SFC) may be performed either in open (capillary) columns or in packed columns. Both approaches have been demonstrated numerous times in the literature. In this contribution it will be attempted to discuss some aspects of columns for SFC. Some advantages of both types of columns will be identified. Attention is paid to the stationary phase film thickness, the speed of analysis, and to the maximum number of theoretical plates (effect of column pressure drop). In this brief contribution many questions will be left unanswered and many significant aspects will be left undiscussed, illustrating that much research remains to be done in this area.     

Monolithic silica rod columns for high-efficiency reversed-phase liquid chromatography

Chromatographic properties of a new type of monolithic silica rod columns were examined. Silica rod columns employed for the study were prepared from tetramethoxysilane, modified with octadecylsilyl moieties, and encased in a stainless-steel protective column with two polymer layers between the silica and the stainless-steel tubing. A 25 cm column provided up to 45,000 theoretical plates for aromatic hydrocarbons, or a minimum plate height of about 5.5 μm, at optimum linear velocity of ca. 2.3 mm/s and back pressure of 7.5 MPa in an acetonitrile-water (80/20, v/v) mobile phase at 40°C. The permeability of the column was similar to that of a column packed with 5 μm particles, with K(F) about 2.4×10(-14) m(2) (based on the superficial linear velocity of the mobile phase), while the plate height value equivalent to that of a column packed with 2.5 μm particles. Generation of 80,000-120,000 theoretical plates was feasible with back pressure below 30 MPa by employing two or three 25 cm columns connected in series. The use of the long columns enabled facile generation of large numbers of theoretical plates in comparison with conventional monolithic silica columns or particulate columns. Kinetic plot analysis indicates that the monolithic columns operated at 30 MPa can provide faster separations than a column packed with totally porous 3-μm particles operated at 40 MPa in a range where the number of theoretical plates (N) is greater than 50,000.     

Evaluation of packed capillary liquid chromatography columns and comparison with conventional-size columns

Apart from extracolumn effects peak dispersion in liquid chromatographic columns is caused by the column inlet, the packed bed, and the column outlet. A strategy applicable for independent evaluation of the individual sources of column band broadening was developed on the basis of the linear extrapolation method (LEM). This method was applied to compare the performance of packed capillary LC columns from various commercial suppliers with conventional-size columns. The columns differed widely in their performance with respect to peak shapes and widths for standard substances. The capillary columns were found well packed, but in some cases overall performance would benefit from improving the design of the area between the packed bed and the connecting capillaries, containing frits as well as dead volumes.     

Hydrodynamic chromatography: packed columns,multiple detectors,and microcapillaries

Hydrodynamic chromatography (HDC) is a liquid chromatographic technique that separates analytes on the basis of their size in solution. Separation can be conducted either in an open tube or in a column packed with inert, nonporous beads. In HDC, larger analytes elute first and smaller ones later, due to preferential sampling of the streamlines of flow in the open tube or in the interstitial medium of the packed column. Because of the low shear rates experienced in HDC, coupled with the wealth of information obtained when employing a multiplicity of detection methods, the technique has experienced a resurgence in recent years in both the particle sizing and macromolecular arenas, where it can provide information on the mutual interdependence of molar mass, size, shape, and compactness. Additionally, microcapillary HDC is also gaining popularity amongst the bioanalytical community, who have employed the technique, inter alia, to separate DNA fragments over a base pair range spanning four orders in magnitude. Here, examples from the literature are used to show how HDC has been applied in each of the aforementioned areas, explaining the information that can be obtained from various detector combinations, and opining on the future of the technique.     

Performance of new prototype packed columns for very high pressure liquid chromatography

The reduced heights equivalent to a theoretical plate (HETP) of naphtho[2,3-a]pyrene were measured at room temperature on two sets of new prototype columns designed to be used in very high pressure liquid chromatography (VHPLC). The mobile phase used was pure acetonitrile. The columns are 50, 100, and 150 mm long. Those of the first set are 2.1 mm I.D., those of the second set, 3.0 mm I.D. The performance of these new columns were compared to those of the first generation of VHPLC columns, commercially available in 2.1 mm I.D. The prototype and commercial columns behave similarly at low reduced linear velocities (ν<5$\nu <5$), when the heat effects are negligible. At high flow rates, the shorter prototype columns have a twice better efficiency and less steep C-branches than the commercial columns. In contrast, the C-branch of the 150 mm long prototype columns are slightly steeper than those of the commercial columns. The important contribution to the reduced HETP that is due to the heat effects at high flow rates can in part be accounted for by a band broadening model governed by a flow mechanism with the shortest prototype columns. The sole heat effects cannot, however, explain the mediocre reduced HETPs of the 2.1 and 3.0 I.D. 150 mm long prototype columns. It seems that radial heterogeneity of the flow rate of the long prototype columns is significantly larger than that of the short columns. The contribution of the packing heterogeneity adds up to that of the heat effects to yield a poor column efficiency when sub-2μm$\text{sub-}2\mu \text{m}$ are packed into thin, long column tubes.     

Gas chromatography of hydrocarbons using packed liquid chromatographic capillary columns

Capillary columns of 0.3-0.5 mm i.d. packed with 3- to 30-μm silica-based stationary phases for liquid chromatography were used for gas chromatographic separation of hydrocarbons. Column efficiencies were evaluated for various commercially available packing material. The best column efficiency was achieved with 5-μm octadecyl group bonded silica gel, the surface of which was coated with a poly (dimethylsiloxane) film. The 30-cm column produced 11,000 theoretical plates.     

Fast supercritical fluid chromatography of polymers using packed capillary columns

Summary Fast separations of perfluorinated polyethers and polymethylsiloxanes that are composed of 50–80 oligomers were demonstrated in packed capillary column supercritical fluid chromatography (SFC) using a carbon dioxide mobile phase. Separations were accomplished within 10 min using a 13 cm×250 μm i.d. column packed with 2 μm porous octadecyl bonded silica (ODS) particles. Effects of particle diameter of the packing material and pressure programming on separation were investigated, and packed column SFC was compared with open tubular column SFC. Results show that as the particle diameter was decreased from 5 to 3 to 2 μm and the column length was reduced from 85 to 43 to 13 cm, the separation time could be reduced from 70 to 20 to 10 min while still maintaining similar separation (resolution). Short columns packed with small porous particles are very suitable for fast SFC separations of polymers.     

A miniature gradient elution system for liquid chromatography with packed capillary columns

A miniaturized continuous gradient elution system was designed for work with packed capillary columns. The retention reproducibility achieved is adequate for many practical applications.     

Preparative high-performance liquid chromatography with reversed phase packed glass columns

An efficient system for preparative reversed-phase separations with packed glass columns is described. The advantage of this system is the use of relatively simple and inexpensive equipment. Column performance, load capacity, effect of the feed volume and the feed concentration on peak broadening are shown. The influence of the selectivity and the capacity factors on column load have been measured. The effect of the column dimensions is demonstrated by means of practical applications. The loading capacity of a column depends on the thermodynamic proporties of the separation system used. It is therefore not expedient in preparative chromatography to correlate the loading capacity of a column by means of grams dissolved per grams of adsorbent.     

Performance and lifetime of slurry packed capillary columns for high performance liquid chromatography

Fused silica capillary columns of the internal diameter of 320 μm were packed with the Nucleosil C18 stationary phase of 5 μm using the slurry packing method. The time of the bed compaction phase, packing pressure, and the use of ultrasound varied to study their influence on the column performance. Van Deemter curves were measured and separation impedance values were calculated in order to assess both separation efficiency and kinetic performance of the columns. Selected columns were tested again after nine months to evaluate the stability of their beds. Separation efficiencies of all columns were similar, but a major difference, caused by the use of ultrasound, was observed in the bed stability. Columns sonicated for 25 minutes during the bed compaction phase exhibited unchanged performance in the course of several months, while the performance of non-sonicated columns decreased.     

Hydrodynamic impact of particle shape in slurry packed liquid chromatography columns

We report on a series of flow velocity and efficiency profiles, which were measured across the cross section of preparative chromatographic columns packed with different stationary phase materials using computed tomography. It is shown that this non-invasive technique is very useful for visualization of the inner part of a packed column and measurement of the spatial resolved column packing properties. For evaluation of the influence of the particle shape on the velocity distribution and column performance, irregular and spherical reversed phases were studied in detail. The results showed a decreasing velocity towards the column wall most certainly due to a lower permeability. This effect was much less pronounced in the case of spherical particles, indicating a more homogenous packing structure. The influence of the column packing pressure, as a possible measure for improvement of the packing homogeneity was also studied. It was shown that under the same packing conditions spherical particles always lead to a more homogeneous packing. The overall results of this work contribute to the origin of the fact that spherical material is superior to irregular one from the hydrodynamic point of view.     

Theoretical and experimental study of packed capillary columns in gas chromatography

Chromatographia - The coupling theories byGiddings andLittle-wood are reviewed and discussed. A practical method to derive the values of the constants involved in the plate height equation from...     

On-line extraction and separation by supercritical fluid chromatography with packed columns

The application of fluid extraction in combination with fluid chromatography with packed column and flame ionization detection is described. Fluid chromatographic equipment is shown. Applications of this system to drug characterization are demonstrated.     

Retention mechanisms in super/subcritical fluid chromatography on packed columns

Whereas the retention rules of achiral compounds are well defined in high-performance liquid chromatography, on the basis of the nature of the stationary phase, some difficulties appear in super/subcritical fluid chromatography on packed columns. This is mainly due to the supposed effect of volatility on retention behaviours in supercritical fluid chromatography (SFC) and to the nature of carbon dioxide, which is not polar, thus SFC is classified as a normal-phase separation technique. Moreover, additional effects are not well known and described. They are mainly related to density changes of the mobile phase or to adsorption of fluid on the stationary phase causing a modification of its surface. It is admitted that pressure or temperature modifications induce variation in the eluotropic strength of the mobile phase, but effects of flow rate or column length on retention factor changes are more surprising. Nevertheless, the retention behaviour in SFC first depends on the stationary phase nature. Working with polar stationary phases induces normal-phase retention behaviour, whereas using non-polar bonded phases induces reversed-phase retention behaviour. These rules are verified for most carbon dioxide-based mobile phases in common use (CO(2)/MeOH, CO(2)/acetonitrile or CO(2)/EtOH). Moreover, the absence of water in the mobile phase favours the interactions between the compounds and the stationary phase, compared to what occurs in hydro-organic liquids. Other stationary phases such as aromatic phases and polymers display intermediate behaviours. In this paper, all these behaviours are discussed, mainly by using log k-log k plots, which allow a simple comparison of stationary phase properties. Some examples are presented to illustrate these retention properties.     

Modifier effects on packed and capillary columns in supercritical fluid chromatography

Summary The separation of polar thermally labile solutes is one of the potentially most rewarding fields of SFC application. A presupposition for such applications is, however, mobile phases having relatively high solvent strengths. A promising approach to achieve this is the use of mobile phases consisting of carbon dioxide with a polar additive. In this work, the chromatographic effects of different concentrations of an additive, isopropanol, in carbon dioxide have been studied on capillary and packed columns. A series of antibiotics was used as test substances. Best results were obtained with carbon dioxide/8% isopropanol as mobile phase on a capillary column coated with a cyanopropyl-substituted polysiloxane stationary phase.