Monolithic silica xerogel capillary column for separations in capillary LC and pressurized CEC.

Monolithic capillary columns were prepared by the reaction of a mixture of potassium silicate solution and formamide. The surface of the monolith was coated with a thin film formed by a sol-gel method to increase the surface area of the monolith and simultaneously covered with C8 as stationary phase for reversed-phase separation. The morphology of the monolithic column was investigated by SEM. Monolithic columns prepared in this manner showed high permeability and can be operated in capillary LC (CLC) mode at a pressure of 20 psi. PAHs were used to evaluate the separation performance of the stationary phase using CLC and pressurized CEC (pCEC). Efficiencies of 20 000 and 28 000 plates per meter for naphthalene were obtained in CLC and pCEC modes, respectively. Improvement in column efficiency and reduction in analysis time over CLC and improvement in operation facility and separation selectivity over CLC were found using pCEC mode.

Gold microspheres modified with octadecanethiol for capillary liquid chromatography.

Monodispersed spherical gold particles synthesized and modified with n-octadecanethiol (C18-Au) were packed into a 100 microm I.D. capillary column and tested in capillary high-performance liquid chromatography (microHPLC). To the best of our knowledge, this represents the first report on the actual use of micron gold particles as stationary phase in a packed column microHPLC. As measured by scanning electron microscopy, the average diameter of these gold microspheres was 3.5 microm and the surface area, average pore diameter, and average pore volume were 49.4m(2)/g, 4.8 nm, and 0.12 cm(3)/g, respectively. The retention behavior of neutral compounds on the n-octadecanethiol-modified gold microspheres was investigated by separating a mixture of small organic compounds using microHPLC. The results from the experiments show that C18-Au behaves basically as a reversed phase. The test of chemical stability of the C18-Au stationary phase under alkaline condition demonstrates that it is stable with the flushing of a mobile phase at pH 12 for at least 140 h. The C18-Au particles are also mechanically strong enough to withstand pressure up to 52 MPa. The preliminary results in this work prove the feasibility of the fabrication of C18-Au micron particles as a novel stationary phase for packed column microHPLC.

Open-tubular gas chromatography using capillary coated with octadecylamine-capped gold nanoparticles.

The octadecylamine-capped gold nanoparticles (ODA-Au-NPs) were prepared and directly used to coat the capillary wall. The hydrophobic coating acted as the stationary phase for open-tubular gas chromatography (OTGC). The ODA-Au-NPs can be adsorbed tightly onto the inner surface of fused silica capillary column via electrostatic interaction and enhanced interaction of van der Waals between gold nanoparticles and the capillary wall. Thus, the modification of the inner surface of capillary column by ODA-Au-NPs can be achieved simply by flushing the capillary with a solution of ODA-Au-NPs and the resulted ODA-Au-NPs coating is very stable. No perceptible degradation in the ODA-Au-NPs-based separation was observed after approximately 1900 sample runs. This type of columns also provided excellent chromatographic performances: high number of theoretical plates, outstanding run-to-run and column-to-column reproducibility, and high selectivity for a wide range of test mixtures. An efficiency of 2474 theoretical plates per meter for chlorobenzene was obtained on an ODA-Au-NPs-modified 1.6 m x 100 microm i.d. fused silica capillary column.

Pressurized capillary electrochromatographic assay of trimethoprim impurities using 1 microm particle-based columns.

One micrometre silica particles, derivatized with C18, were electrokinetically packed into a 75-microm-i.d. capillary. The resulting column was evaluated for the separation of trimethoprim (TMP) and its impurities using pressurized capillary electrochromatography (pCEC), starting from a capillary liquid chromatographic (CLC) separation. These samples require gradient elution when separated by high performance liquid chromatography (HPLC), but with the new columns isocratic elution suffices for their separation by CLC or pCEC. Only 70,000 theoretical plates/m for impurity C were achieved using CLC mode at relative low pressure (78 bar) although very small particles were utilized. When a voltage above 2 kV (50 V/cm) was applied, unknown peaks appeared, which was assumed due to an electrophoretic effect with the unknown peaks resolving as a result of the applied voltage. In order to minimize these unfavorable contributions, only a low voltage was applied, still leading to higher separation performances and shorter separation times than in CLC. The optimal analyzing conditions in pCEC included a pressure of 78 bar, an applied voltage of 1 kV, and a mobile phase consisting of 80 mM sodium perchlorate (pH 3.1)/methanol (60/40, v/v). These conditions were used to separate and quantify four major impurities in TMP within 22 min. The obtained calibration curves were linear (r>0.9980) in concentration ranges between 0.005 and 0.1 mg/mL for impurities A and C; 0.02 and 0.10 mg/mL for impurity F; and 0.01 and 0.10 mg/mL for impurity H. The detection limits (S/N=3) for impurities A, C, F, and H were 0.52, 0.84, 3.18, and 2.41 microg/mL, respectively. The calibration curves were successfully applied to analyze spiked bulk samples, with mean recoveries ranging from 92% to 110%. The developed method can therefore be considered simple, rapid, and repeatable.

The simultaneous separation and determination of six flavonoids and troxerutin in rat urine and chicken plasma by reversed-phase high-performance liquid chromatography with ultraviolet-visible detection.

The method of high-performance liquid chromatography (HPLC) with UV-vis detection was used and validated for the simultaneous determination of six flavonoids (puerarin, rutin, morin, luteolin, quercetin, kaempferol) and troxerutin in rat urine and chicken plasma. Chromatographic separation was performed using a VP-ODS column (150 mm x 4.6 mm, 5.0 microm) maintained at 35.0 degrees C. The mobile phase was a mixture of water, methanol and acetic acid (57:43:1, v/v/v, pH 3.0) at the flow rate of 0.8 mL/min. Six flavonoids and troxerutin were analyzed simultaneously with good separation. On optimum conditions, calibration curves were found to be linear with the ranges of 0.10-70.00 microg/mL (puerarin, rutin, morin, luteolin, quercetin, kaempferol) and 0.50-350.00 microg/mL (troxerutin). The detection limits were 0.010-0.050 microg/mL. The method was validated for accuracy and precision, and it was successfully applied to determine drug concentrations in rat urine and chicken plasma samples from rat and chicken that had been orally administered with six flavonoids and troxerutin.

Trace measurement of phenothiazine drugs in tablets by micellar-enhanced fluorophotometric method.

It was found that in buffer solution of pH 7.0, the addition of sodium dodecyl sulfate (SDS) to the solution of phenothiazine drugs, such as chlorpromazine, promethazine and trifluoperazine, showed a remarkable enhancement of their fluorescence intensity. A further study proved that the phenothiazine drugs can be determined by fluorophotometric method in micellar system. Under optimal conditions, there was a good linear relationship between fluorescence intensity and phenothiazine compounds concentration, and the detection limit of 3.0 x 10(-8) M chlorpromazine, 3.0 x 10(-8) M promethazine and 1.5 x 10(-8) M trifluoperazine (S/N=3) were also obtained. This method has been used to determine phenothiazine drugs in tablets with satisfactory results.

Electrochromatography with a 2.7 mm inner diameter monolithic column.

Monolithic columns of 2.7 mm I.D. have been prepared and used in electrochromatography (EC) separation. Although capillary electrochromatography (CEC) has higher separation efficiency, it displays some shortcomings, such as limited sample loadability and restricted concentration detectability etc. In this paper, we investigate the feasibility of EC separation with millimeter diameter monolithic columns. By using a designed preparation method of monolithic column packed with about 150 microm quartz sand, the effect of Joule heating can be reduced, and the processes of frit making and column packing can be avoided. The concentration detectability of the EC is improved comparing with that of CEC. Moreover, the separation efficiency of 52,000 plates/m was achieved with a 70 mm length and 2.7 mm I.D. monolithic column.

Stacking ionizable analytes in a sample matrix with high salt by a transient moving chemical reaction boundary method in capillary zone electrophoresis.

The paper presents a novel on-line transient moving chemical reaction boundary method (tMCRBM) for simply but efficiently stacking ionizable analytes in high-salt matrix in capillary zone electrophoresis (CZE). The powerful function and stability of the tMCRBM are elucidated with the ionizable test analytes of L-phenylalanine (Phe) and L-tryptophan (Trp) in the matrix with 85.6-165.6 mM sodium ion and further compared with the normal CZE of Phe and Trp samples dissolved in running buffer. The results verify that (1) the on-line tMCRBM mode can evidently increase separation efficiency, peak height, and resolution, (2) with the mode, the analytes in a 28-cm high-salt matrix plug can be stacked successfully and further separated well, (3) the values of relative standard deviation of peak height, peak area, and migrating time range from 3.9% to 6.1%; the results indicate the high stability of the technique of tMCRBM-CZE. The techniques implies obvious potential significance for those ionizable analytes, e.g., protein, peptide, and weak alkaline or acidic compound, in such matrixes as serum, urine, seawater, and wastewater, with high salt, which has a deleterious effect on isotachophoresis (ITP) and especially on electrostacking and field-amplified sample injection (FASI). The mechanism of stacking of zwitterionic analytes in a high-salt matrix by the tMCRBM relies on non-steady-state isoelectric focusing (IEF) but not on transient ITP, electrostacking, and FASI.

Improving separation efficiency of capillary zone electrophoresis of tryptophan and phenylalanine with the transient moving chemical reaction boundary method.

A simple and convenient mode--moving chemical reaction boundary method-capillary zone electrophoresis (MCRBM-CZE)--was designed for the enhancement of separating efficiency of CZE. In this mode, the transient MCRBM is used for the on-line pre-treatment of sample. By analyses of tryptophan (Trp) and phenylalanine (Phe) as an example, the experiments by MCRBM-CZE were carried out and further compared with those by normal CZE without the transient MCRBM. The results reveal that by carefully selected appropriate electrolytes, a strong condensation effect can be achieved by using MCRBM-CZE; this effect can greatly improve the separation efficiency, resolution and peak height of Trp and Phe in CZE as compared with those of normal CZE of Trp and Phe. Even if the sample comprises high concentrations of salt, such as 80 mM NaCl (concentration of sodium ion up to 145.6 mM), the same condensation effect can also been observed; this implies obvious significance for biological samples like urine and serum. However, if the electrolytes was chosen inappropriately only a poor compression effect of sample was observed in the MCRBM-CZE runs.