High Retention Factors Research Articles

Preparation of carboxyl-functionalized silica core-shell microspheres and their applications in weak cation exchange chromatography, heavy metal removal, and lysozyme enrichment.

Chromatography is a technique of separation based on adsorption and/or interaction of target molecules with stationary phases. Herein, we report the design and fabrication of BTDA@SiO2 core-shell microspheres as a new class of stationary phase and demonstrate its impressive performance for chromatographic separations. The silica microspheres of BTDA@SiO2 were synthesized by in situ method with 1,3,5-benzenetricarboxaldehyde and 3,5-diaminobenzoic to separate peptides and proteins on high-performance liquid chromatography. The BTDA@SiO2 core-shell structure has a high specific surface area and retention factor of 4.27 and 8.31 for anionic and cationic peptides, respectively. The separation factor and resolution were high as well. A typical chromatogram illustrated nearly baseline resolution of the two peptides in less than 3 min. The BTDA@SiO2 was also highly stable in the pH range of 1 to 14. Furthermore, the prepared BTDA@SiO2 core-shell material not only be used for chromatographic separation but also as heavy metal removal from water. Using a BTDA@SiO2, we also achieved a lysozyme enrichment with a maximum saturated adsorption capacity reaching 714mg/g. In summary, BTDA@SiO2 has great application prospects and significance in separation and purification systems.

Retention behavior of nucleosides and nucleobases on a 3 μm undecylenic acid-functionalized silica column in per aqueous liquid chromatography and hydrophilic interaction liquid chromatography separation modes

A 3 μm undecylenic acid-functionalized stationary phase (UAS) was prepared for the separation of nucleosides and nucleobases using per aqueous liquid chromatography (PALC) and hydrophilic interaction liquid chromatography (HILIC). The retention behaviors of nucleosides and nucleobases in PALC and HILIC modes were explored by adjusting parameters such as water content, buffer concentration, pH of the mobile phase and column temperature. The experimental data and separation chromatogram demonstrated that PALC could provide retention comparable to that of HILIC for nucleosides and nucleobases. Comparative studies using diluted adenosine solutions evaluated theoretical plates and peak shape for the same retention factors (between 0.25 and 5.0) in PALC and HILIC. There was no buffer component in the mobile phases used to operate the comparisons. HILIC mode is more efficient for adenosine than PALC mode at low retention factors. It's the exact opposite phenomenon for high retention factors. It is proposed that the mass transfer of adenosine between the UAS, the water-rich layer and the ACN-rich mobile phase in HILIC is relatively slow. Given the significant use of toxic ACN in HILIC, PALC emerges as a safer and more effective alternative for separating nucleosides and nucleobases.

On the contribution of the top and bottom walls in micro-pillar array columns and related high-aspect ratio chromatography systems

We report on a steady-state based, and hence highly accurate numerical modelling study of the effect of the top and bottom wall in the current generation of micro-pillar array columns. These have a mesoporous retention layer that not only covers the pillar walls but also the bottom wall. Our results show that the performance of these columns can in general not be improved by also covering the top wall with the same layer, despite the increased column symmetry this approach would offer. The reason for this is that the local species retardation caused by a retentive layer is much stronger than the pure flow arresting effect of an uncovered wall. At least, this has a crucial impact in high aspect-ratio systems such as micro-pillar array columns because these require a small inter-pillar distance to promote mass transfer together with a large channel depth to enable a sufficiently high flow rate. On the other hand, a notable improvement could be made if micro-pillar array would be produced without having a retentive layer at the bottom. At Péclet number Pe = 50 and aspect ratio AR = 5 for flow-channels, this gain amounts up to about 4.5 h-units at a zone retention factor k’’ = 2 and 1.75 h-units at k’’ = 16 (gain scales almost linearly with Pe).To verify these results, we also considered another high aspect-ratio system with a simplified geometry: the open-tubular channel with a flat-rectangular cross-section. This led to very similar observations, thus confirming the findings for the micro-pillar array.The results produced in the present study also allow us to conclude that the classic modelling paradigm adopted in chromatography, which is based on the independency and hence additivity of the hCm- and hCs-contributions, can lead to large modelling errors in chromatographic systems with a high aspect-ratio, even when their geometry is so simple as that of a straight open-tubular channel with constant cross-section. Indeed, when both zones are treated independently, the analysis misses how the vertical diffusion through the retentive layer helps suppressing the vertical gradients in the mobile zone. The diffusion through this layer occurs in a ratio of k’’Ds/Dm (Dm being the diffusion coefficient in mobile phase zone and Ds being the diffusion coefficient in stationary phase zone), such that at high retention factors this diffusion contribution even becomes the dominant one.

Silver-doped graphite oxide composites used as antimicrobial agents against Staphylococcus aureus, Escherichia coli and Tatumella terrea evaluated by direct TLC bioautography

Silver-doped graphite oxide composites presented mixed Ag and Ag2O crystalline phases accompanied by C/O and ID/IG average ratios of 2.13 and 1.16, associated with a good oxidation degree and high structural disorder or defects in the carbon material, respectively. The low-silver-doped GrO composite showed homogenous silver particle dispersion and a low particle size distribution (96 nm). However, high-silver-doped GrO composites generated materials with high relative crystallinity, silver particle agglomeration, and Ag metal phase promotion. At 3 and 5 mg ml−1 per chromatographic plate, the silver-doped graphite oxide composites were tested by direct TLC bioautography against Staphylococcus aureus, Escherichia coli, and Tatumella terrea microorganisms, where the minimum inhibitory concentration was 3 mg ml−1 per chromatographic plate. At 3.0 mg ml−1 per chromatographic plate, high-silver-doped GrO composites exhibited a 39%, 3.2-fold, and 83% higher retention factor (R f ) compared with the composites with low-silver-doped GrO composites against S. aureus, E. coli, and T. terra microorganisms, respectively. However, both composites showed similar inhibition capacities at 5.0 mg ml−1 per chromatographic plate against the three microorganisms. This behavior may be associated with both composites reaching the threshold limit. In general, the silver acetate amount used in the silver-doped GrO composites influenced the dispersion, crystalline phase promotion, particle size distribution, and the silver particle release capacity, which modified the electrostatic adsorption type between the composites and the bacterial cell walls.

Drying methods effect on bioactive compounds, phenolic profile, and antioxidant capacity of mango powder

The effects of various drying methods (spray drying, cabinet drying, vacuum drying, tunnel drying, rotary oven drying, and gas oven drying) on the phenolic profile, bioactive compounds, and antioxidant capacity of mango powder have been considered with the aim of discover the effective drying technique. To identify and quantify the phenolic profile, the Folin-Ciocalteu method was used, while 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging studies were utilized to elucidate the antioxidant capacity. Results showed that total phenolic content (TPC), total flavonoid content (TFC), total anthocyanin content (TAC) and total carotenoid content (TCC) showed substantial variations (p < 0.05) between different drying methods, and the antioxidant properties were influenced to different degrees by drying methods. The antioxidant capacity of dried mango powder is strongly correlated with phenolic compounds (polyphenol, flavonoids, and anthocyanin). Caffeic acid was predominant among the 16 phenolics in mango powder. Flavonoids had higher retention factors (RF) than phenolic acids. Finally, our findings indicate that both cabinet drying and vacuum drying are appropriate for mango powder production. However, taking into account both economic viability and phenolic compound quality, cabinet drying would be the supreme choice for the outturn of mango powder as antioxidant-rich flavor enhancers.

Performance of functionalized monolithic silica capillary columns with different mesopore sizes using radical polymerization of octadecyl methacrylate

We report on the possibility to enhance the phase ratio and retention factor in silica monoliths. According to pioneering work done by Núñez et al. [1], this enhancement is pursued by applying a stationary phase layer via radical polymerization with octadecyl methacrylate (ODM) as an alternative to the customary octadecylsilylation (C18-derivatization). The difference in band broadening, retention factor and separation selectivity between both approaches was compared. Different hydrothermal treatment temperatures for the column preparation were applied to produce monolithic silica structures with three different mesopore sizes (resp. 10, 13, and 16 nm, as determined by argon physisorption) while maintaining similar domain size (sum of through-pore and skeleton size). It has been found that the columns with the poly(octadecyl methacrylate)-phase (ODM columns) provided a 60 to 80% higher retention factor in methanol-water mixture compared to the octadecylsilylated (ODS) columns produced by starting from similar silica backbone structures. In acetonitrile-water mixture, the enhancement is smaller (15 to 30% times higher), yet significant. By adjusting the fabrication conditions (for both the preparation of the monolithic backbones and the surface functionalization), the achieved retention factors (up k = 4.89 for pentylbenzene in 80:20% (v/v) methanol/water) are obviously higher than obtained in the pioneering study on ODM monoliths of Núñez et al. [1], and column clogging could be completely avoided. In addition, also separation efficiencies were significantly higher than shown in Ref. [1], with plate heights as low as 5.8 μm. These plate heights are however inferior to those observed on the ODS-modified sister columns. The difference can be explained by the slower intra-skeleton diffusion displayed by the ODM-modified columns, in turn caused by the larger obstruction to diffusion originating from the thicker stationary phase layer.

Recovery of cadmium, lead and nickel from leach solutions of waste electrical and electronic equipment using activated carbon modified with 1-(2-pyridylazo)-2-naphthol

Waste electrical and electronic equipment (WEEE) are considered as a major source for both virtuous and hazardous metals. Hazardous metals make their way to cause pollution of soil and water. The risk of contamination is very high where WEEE is being processed. In the present study activated carbon was synthesized from sawdust and modified with 1-(2-pyridylazo)-2-naphthol (PAN) through diazotization. The surface modification was confirmed through various instrumental analysis like CHNO elemental analyzer, FTIR, TGA/DSC, BET, SEM and EDX. Surface area of activated carbon calculated using BET method was 178 m2/g with average pore size of 14.5 (A°) however, surface area gets reduce to 68 m2/g after surface modification. The synthesized adsorbent was packed in a column and used for extraction of cadmium, lead and nickel from WEEE and some soil samples. Different analytical parameters like pH, sample volume and flow rate, eluent type and flow rate, effect of coexisting ions and initial concentration were optimized. It was observed that the adsorbent has high retention factor (> 94%) for the selected metal ions at neutral pH with 12 mL/min sample flow rate. Nitric acid solution (5 mol/L) of 7 mL was found suitable for quantitative elution with flow rate of 2 mL/min. The method was efficient even with higher sample volume of 1000 mL, 1200 mL and 900 mL with enrichment factor of 143, 171 and 129 for Cd, Pb and Ni, respectively. As compared to batch adsorption, cartridge of the synthesized adsorbent can be regenerated and used multiple times with ease. Similarly, it also prevents both loss of adsorbent and collected metals ions as normally in case of filtration or centrifugation process.

Structure-Directed Screening and Analysis of Thyroid-Disrupting Chemicals Targeting Transthyretin Based on Molecular Recognition and Chromatographic Separation.

Exposure to thyroid-disrupting chemicals (TDCs) poses a great threat to human health. However, the screening and analysis of TDCs in environmental samples remain a tough work. In this study, we reported a structure-directed strategy for analyzing TDCs targeting transthyretin (TTR) based on molecular imprinting and chromatographic separation. The imprinted composites were prepared using l-thyroxine (T4) as a template and a tryptophan-like monomer screened from the amino acid library. The imprinted composites exhibited an adsorption capacity of 22.2 μmol g-1 for T4 and an imprinting factor of 2.1. Chromatographic testing was then conducted among 72 chemicals using the imprinted composites-packed column. High retention factors were observed for chemicals that were structurally similar to T4. The chromatographic results were compared with a data set of 45 chemicals with known activities toward TTR. The results suggested that chemicals can be distinguished as TTR binders and nonbinders by retention factors, with a predictive accuracy of more than 90%. Moreover, the retention factors of chemicals were highly correlated with the reported relative potencies obtained from TTR assays. Thus, screening of TTR-binding chemicals can be realized through this simple chromatographic method. The imprinted composites were applied for target analysis and nontarget analysis of TTR-binding chemicals in dust samples. Three new TTR binders were successfully identified and verified by this method. The combination of molecular imprinting and chromatography opens up a new approach for screening TDCs targeting TTR.

Transient Ion-Pair Separations for Electrospray Mass Spectrometry.

We report a novel ion-pair chromatography (IPC) approach for liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS), where the eluent does not contain any ion-pairing reagent (IPR). The IPR is injected on the column, much like the sample, and moves down the column. Significant amounts of a high retention factor IPR is injected, resulting in a transient but reproducible regional coating that progresses along the column. The sample is injected after a brief interval. The sample components interact with the IPR coated region during their passage; the chosen eluent gradient elutes the analytes of interest into the mass spectrometer before the IPR. Following analyte elution, the gradient is steeply raised, the IPR is washed out, and the effluent is sent to waste via a diverter valve until it is fully removed. As the nature of the analyte retention continuously changes along the column and with time, we call this transient ion-pair separation (TIPS). As the IPR never enters the MS, TIPS addresses two major drawbacks of IPC for ESI-MS: it avoids both ion suppression and ion source contamination. The potential of the generic approach for other modes of separation is discussed. An illustrative separation of two small inorganic ions, iodate and nitrate, is demonstrated on a reverse phase column by a transient prior injection of hexadecyltrimethylammonium chloride as IPR.

Analyses of Total Alkaloid Extract of Corydalis yanhusuo by Comprehensive RP × RP Liquid Chromatography with pH Difference.

A comprehensive two-dimensional (2D) reverse phase (RP) liquid chromatography (LC) method is developed for alkaloid analysis. This offline comprehensive 2D method is developed using different pH values. With a pH value of 10.5, most alkaloids appear in the form of neutral molecules possessing high retention factors based on their polarity, while the alkaloid polarity order is changed when the pH value decreased to 3.0. The performance of pH modulated 2D LC is demonstrated with 8 alkaloid standards which resulted in orthogonal separation. The developed method is then applied to total alkaloid separation in Corydalis yanhusuo. The first-dimension separation is carried out using methanol and water containing 1.0% ammonium hydroxide and a strong base-resistant RP column, which afforded a peak capacity of 94. The second-dimension analysis is carried out with a surface positive charge column providing a peak capacity of 205 using a mobile phase consisting of acetonitrile and water with 0.15% formic acid. 2D analyses of total alkaloid extract from C. yanhusuo afford a total peak capacity of 9090. Sixteen compounds were tentatively identified based on their ultraviolet spectrum and MS/MS analyses. The proposed method provides an alternative approach to achieve high peak capacity for analysis of alkaloid extract.

On the inherent data fitting problems encountered in modeling retention behavior of analytes with dual retention mechanism

Some valuable insights have been obtained in the inherent fitting problems when trying to predict the retention time of complex, multi-modal retention modes such as encountered in HILIC and SFC. In this study, we used mathematical models with known input parameters to generate different sets of numerical test curves representative for systems exhibiting a complex, non-LSS dual retention behavior. Subsequently, we tried to fit these data sets using some popular (non-linear) literature models. Even in cases where a physical fitting model exists (e.g., the mixed model in case of pure additive adsorptive and partitioning retention), the fitting quality can only be expected to be relatively good (prediction errors expressed in terms of a normalized resolution error ɛRs) when carefully selecting the scouting runs and the appropriate starting values for the fitting algorithm. The latter can best be done using a comprehensive grid search scanning a wide range of different starting values. This becomes even more important when no good physical model is available and one has to use a non-physical fitting model, such as the empirical Neue-model. The use of higher-order models is found to be quasi indispensable to keep the prediction errors on the order of some ΔRs=0.05. Also, the choice of the scouting runs becomes even more important using these higher-order models. For highly retained compounds we recommend using scouting runs with long tG/t0-values or to include a run with a higher fraction of eluting solvent at the start of the gradient. When trying to predict gradient retention, errors with which the isocratic retention behavior is fitted are much less important for high retention factors k than errors made in the range of k near the one at the point of elution. The results obtained with a so-called segmented Neue-model (containing 7 parameters) were less good and thus practically not interesting (because of the high number of initial runs).

Modeling full-scale osmotic membrane bioreactor systems with high sludge retention and low salt concentration factor for wastewater reclamation

A full-scale model was developed to find optimal design parameters for osmotic membrane bioreactor (OMBR) and reverse osmosis (RO) hybrid system for wastewater reclamation. The model simulates salt accumulation, draw solution dilution and water flux in OMBR with sludge concentrator for high retention and low salt concentration factor. The full-scale OMBR simulation results reveal that flat-sheet module with spacers exhibits slightly higher flux than hollow-fiber; forward osmosis (FO) membrane with high water permeability, low salt permeability, and low resistance to salt diffusion shows high water flux; an optimal water recovery around 50% ensures high flux and no adverse effect on microbial activity; and FO membrane cost decreases and RO energy consumption and product water concentration increases at higher DS flow rates and concentrations. The simulated FO water flux and RO energy consumption ranges from 3.03 to 13.76LMH and 0.35 to 1.39kWh/m3, respectively.

Comparison of the kinetic performance and retentivity of sub-2 μm core–shell, hybrid and conventional bare silica phases in hydrophilic interaction chromatography

The separation performance and retention properties of four sub-2μm underivatised silica materials were evaluated in the hydrophilic interaction chromatography (HILIC) mode. These included an organic/inorganic hybrid silica, conventional silica, narrow particle size distribution silica and a core–shell silica. Van Deemter characterisation was performed using conditions to give high retention factors (k=5.5–6.0) with 10cm columns to limit the contribution of extra-column dispersion. The core–shell 1.6μm bare silica (Cortecs) was shown to be kinetically superior to fully porous particle types. Little column-to-column variation in the reduced b-coefficient was observed for the test analytes as corroborated by arrested elution experiments. However, the reduced b-coefficient was shown to be different between analytes, e.g. cytosine versus nortriptyline. It is speculated that the nature of the retention mechanism (hydrophilic versus ionic retention) and solute physiochemical properties perhaps influence the b-coefficient. Maxwell-Effective Medium Theory (EMT) applied to results for a wider range of solutes indicated that the intra-particle diffusion (Dpart) behaviour for individual compounds is broadly similar irrespective of the particle morphology in HILIC. Finally, the impact of varying buffer concentration for a test mix showed that retention and peak shape varied considerably between different silicas. High efficiency separations can be achieved for hydrophilic and basic solutes using a combination of sub-2μm core shell bare silica particles and appropriate buffer concentrations.

Application of CO2-Saturated Water Flooding as a Prospective Safe CO2 Storage Strategy

Abstract In this study, According to the results of CO2 solubility measurement tests at constant temperatures, an increase in CO2 solubility values was observed for CO2–brine and CO2–oil systems when the equilibrium pressure increases. Furthermore, it was revealed that for both aforementioned systems, the solubility of CO2 reduces when temperature increased. It was found that introducing CO2 to the oil reservoirs through injection water provides great opportunity to lock large quantity of CO2 inside the porous medium with high retention factor. Results of this study showed that both secondary and tertiary scenarios of CO2-saturated water flooding are favourable with the storage capacity between 34% to 45% of the injected CO2 in the sand-pack model. In addition, the solubility trapping was shown to be the main CO2 trapping mechanism during CO2-saturated water flooding. According to the results of this study, most of the injected CO2 was trapped in the porous medium through dissolution in the reservoir fluids, i.e., solubility trapping. As an example, the flooding tests conducted in secondary mode at P = 10.3 MPa and T = 25 °C, showed that about 95% of the injected CO2 can be trapped by dissolution mechanism into the reservoir oil and brine. In terms of oil recovery, it was found that the ultimate oil recovery factor of CO2-saturated water flooding is consistently more than that of conventional water flooding leading this technique to be a more viable option as a means of improved oil recovery technique. In this study, flooding tests conducted at pressure of P = 10.3 MPa and temperature of T = 25 °C, verified that injection of CO2-saturated water resulted in improving the conventional water flooding oil recovery factor by about 19.0% and 12.5% of OOIP for secondary and tertiary scenarios, respectively. Based on the results obtained in this work, it was concluded that mixing CO2 with injected water noticeably provides permanent, safe, and practical CO2 storage together with considerable oil recovery improvement in light oil systems.

Robust analysis of the hydrophobic basic analytes loratadine and desloratadine in pharmaceutical preparations and biological fluids by sweeping—cyclodextrin-modified micellar electrokinetic chromatography

The analysis of hydrophobic basic analytes by micellar electrokinetic chromatography (MEKC) is usually challenging because of the tendency of these analytes to be adsorbed onto the inner capillary wall in addition to the difficulty to separate these compounds as they exhibit extremely high retention factors. A robust and reliable method for the simultaneous determination of loratadine (LOR) and its major metabolite desloratadine (DSL) is developed based on cyclodextrin-modified micellar electrokinetic chromatography (CD-MEKC) with acidic sample matrix and basic background electrolyte (BGE). The influence of the sample matrix on the reachable focusing efficiency is studied. It is shown that the application of a low pH sample solution mitigates problems associated with the low solubility of the hydrophobic basic analytes in aqueous solution while having advantages with regard to on-line focusing. Moreover, the use of a basic BGE reduces the adsorption of these analytes in the separation compartment. The separation of the studied analytes is achieved in less than 7min using a BGE consisting of 10mmolL−1 disodium tetraborate buffer, pH 9.30 containing 40mmolL−1 SDS and 20mmolL−1 hydroxypropyl-β-CD while the sample solution is composed of 10mmolL−1 phosphoric acid, pH 2.15. A full validation study of the developed method based on the pharmacopeial guidelines is performed. The method is successfully applied to the analysis of the studied drugs in tablets without interference of tablet additives as well as the analysis of spiked human urine without any sample pretreatment. Furthermore, DSL can be detected as an impurity in LOR bulk powder at the stated pharmacopeial limit (0.1%, w/w). The selectivity of the developed method allows the analysis of LOR and DSL in combination with the co-formulated drug pseudoephedrine. It is shown that in CD-MEKC with basic BGE, solute–wall interactions are effectively suppressed allowing the development of efficient and precise methods for the determination of hydrophobic basic analytes, whereas the use of a low pH sample solution has a positive impact on the attainable sweeping efficiency without compromising peak shape and resolution.

Signal enhancement by trapping in microscale liquid chromatography: Numerical modelling

We report on a series of computer simulations that have been made to explore the operation and performance limits of a periodically heated and cooled trapping segment (liquid analog of the modulator device used in GC×GC) interfacing the separation column and the detector. The initial peak width and the retention of the molecules on the trapping segment have the most important influence on the trapping efficiency. Higher retention of the trapping segment and smaller peaks will lead to higher signal enhancements. However when the resulting concentration gradients become too large, as is the case for very small peaks and/or for very high retention factors on the trapping segment, the dispersion will strongly decrease the focussing efficiency. The molar diffusion coefficient and the linear velocity mainly have an impact on the dispersive behaviour, which can be directly calculated from the associated plate height values. General design rules for the trapping segment, validated with the computer simulations, give a good estimate of the required trapping time and the length of the trapping segment. Equations for the estimation of the trapped peak width and signal enhancement are also given.

A numerical study of the assumptions underlying the calculation of the stationary zone mass transfer coefficient in the general plate height model of chromatography in two-dimensional pillar arrays

The present study investigates the validity of one of the key assumptions underlying the general plate height model of chromatography, i.e., the presumed independency of the individual band broadening contributions. More precisely, it is investigated under which conditions the mass transfer inside the stationary zone (e.g., porous pillars) is independent from the axial transport of species outside this zone, and how strongly any such dependency would affect the validity of the general plate height model of chromatography. For this purpose, detailed calculations of the species concentration distribution inside and outside the porous pillars of a computer-mimic of a porous pillar array column have been made. These simulations revealed a clear interplay between the mass transfer inside and outside the pillars, manifesting itself as an asymmetry of the species concentration distribution inside the pillars. The latter is in disagreement with the basic assumption used to calculate the value of the C s-term of the general plate height model. The asymmetry-effect is largest at low reduced velocities, high retention factors and high intra-pillar diffusion coefficients. Fortunately, these are conditions where the C s-term is relatively small, which might explain why the general plate height model of chromatography (and based on the symmetry assumption) can represent the band broadening in a porous pillar array within an accuracy on the order of some 1–2%.

Efficiency of the same neat silica column in hydrophilic interaction chromatography and per aqueous liquid chromatography

The dependencies on the mobile phase flow velocity of the efficiency of a column packed with shell particles of neat porous silica (Halo) was measured under two different sets of experimental conditions. These conditions corresponded to the retention mechanisms of per aqueous liquid chromatography (PALC) at low acetonitrile concentrations and of hydrophilic interaction chromatography (HILIC) at high acetonitrile concentrations. The results are compared. Small amounts of a diluted solution of caffeine were injected in order to record the chromatograms under strictly linear conditions. These efficiencies were measured in both water-rich (PALC retention mechanism) and acetonitrile-rich (HILIC mechanism) mobile phases for the same retention factors, between 0.25 and 2.5. The mobile phases were mixtures of acetonitrile and water containing neither supporting salt nor buffer component. At low retention factors, the efficiency of caffeine is better in the PALC than in the HILIC mode. For k ′ = 0.5 , the minimum reduced height equivalent to a theoretical plate (HETP) is close to 2.5 in PALC while it exceeds 5 in HILIC. The converse is true for high retention factors. For k ′ > 2.5 , the HETP is lower in HILIC than in PALC, because the major contribution to band broadening and peak tailing in this latter mode originates from the heterogeneous thermodynamics of retention and eventually restricts column performance in PALC. Most interestingly, the reduced HETP measured in HILIC for caffeine never falls below 4. This suggests that the mass transfer of caffeine between the multilayer adsorbed phase (due to the interactions of the strong solvent and the silanol groups) and the acetonitrile-rich bulk eluent is slow.

Multi-wall carbon nanotubes bonding on silica-hydride surfaces for open-tubular capillary electrochromatography

Prepared multi-wall carbon nanotube (MWNT) materials, including untreated MWNT, HNO 3-treated MWNT and HNO 3-HCl-treated MWNT were covalently attached onto a silica-hydride-modified capillary by hydrosilation, using the abundant double bonds between the pentagon carbons in the MWNT structure. These MWNT-incorporated capillaries were characterized by SEM, ATR-IR and electroosmotic flow (EOF) measurements in phosphate buffers with a pH range of 3.7–9.3 and in the mixtures of acetonitrile modifier. The untreated capillary was assumed to carry some carboxylate groups formed on the non-acid-treated MWNTs, as it had higher EOF values than the hydride capillary. As the MWNTs were treated with HNO 3 and HCl solutions, the capillaries had increasingly higher EOF values. To examine the existence of an electrochromatography mechanism in the modified capillaries, a mixture of nucleosides and thymine was probed to check the velocity factor and retention factor. In addition to the π–π interaction between the probe solutes and the MWNT immobilized stationary phases; a reversed-phase mechanism could contribute to the chromatographic retention. For acidic tetracyclines, increasing the loadability of MWNTs resulted in a high retention factor and improved the separation resolution.

Detection issues in two-dimensional on-line chromatography

The dilution of analytes during their migration through two-dimensional liquid chromatographic systems was investigated. Simplified equations for the calculation of the dilution factors in the first and second dimension columns were derived considering the variance of the injected sample, the flow rates, the split ratio, the volume of the intermediate sample loop and the sampling time of the first-dimension effluent. The net dilution factor is the product of the dilution factors in the two single dimensions. It is between 200 and 300 in typical two-dimensional liquid chromatographic (2D-LC) separations. These values are less pessimistic than those reported previously [1]. It was shown that the fraction of organic modifier or stronger eluent component in the injected sample has a significant effect on the widths and heights of the eluted peaks. Analytes with high retention factors and high molecular weights are less sensitive toward this effect than those having small retention factors. These results suggest that the optimization of the experimental conditions of two dimension separations must be made from the points of view of both the retention factor and the detector response factor. The frequency of sampling of the eluent of the first-dimension separation does not have a significant effect on the detection limit of a 2D system if the split ratio is adjusted in the ratio of the frequency of sample collection.