High Number Of Theoretical Plates Research Articles

Developed and Validated for the Estimation of Bupropion and Dextromethorphan in a Fixed Dose Combination of the Tablet.

The aim of this study was to develop a simple, accurate, and precise method for the estimation of bupropion and dextromethorphan in a fixed-dose combination of tablets and robust high-performance liquid chromatography for assay analysis of such a fixed combination. Chromatographic analysis was performed and separations were achieved on a Denali C18 150 × 4.6 mm, 5 micron using a mobile phase composition of ortho-phosphoric acid and acetonitrile in the ratio of 600:400 (v/v), flow rate of 1.0 mL/min, injection volume is 10 µL and run time of 6 min in isocratic elution. Ultraviolet (UV) detection was performed at a wavelength of 221 nm. The temperature was maintained at 30 °C. Well-resolved peaks were observed with a high number of theoretical plates, lower tailing factor, and reproducible relative retention time. The method was validated, and all validation parameters were found to be within the acceptance limits. A simple, accurate, and precise method has been developed for estimating bupropion and dextromethorphan in a fixed dose combination of tablets. The optimized method included the following parameters: column temperature of 30 °C, 40% acetonitrile as the mobile phase, and flow rate of 1.0 mL/min. Retention times were 2.25 min and 3.12 min for bupropion and dextromethorphan, respectively. The method was found to be linear in the range of 17.5-105 µg/mL [for R2 < 0.999) and 7.5-45 µg/mL (for R2 > 0.999] for bupropion and dextromethorphan, respectively. Both active pharmaceutical ingredients dissolved more than 90% within 5 min. The current study describes a new, simple, reliable, and economical elution reversed-phase high performance liquid chromatography method for estimating bupropion and dextromethorphan in a fixed combination tablet dosage form. The forced degradation studies were conducted using several degradation conditions such as acidic, alkali, oxidation, thermal, UV, and neutral conditions; the proposed method was effectively employed from the resolution of sample peaks. To the best of our knowledge, no such detailed and stability-indicating method has been reported for a fixed tablet dosage form.

Developed and Validated for the Estimation of Tapinarof in Topical Formulation and Active Pharmaceutical Ingredients.

Its broad applicability and capacity to separate numerous components in a single chromatographic run led to the initial recognition of reversed-phase (RP)-HPLC as an analytical technique. The objective of this study was to create a straightforward and reliable method for accurately and precisely measuring the amount of tapinarof in both the topical formulation and the active pharmaceutical ingredient. Additionally, a robust HPLC assay was developed specifically for analyzing the topical formulation. In this study, chromatographic analysis was conducted using a Kromosil C18 column with dimensions of 250 × 4.6 mm and a particle size of 5 microns. The mobile phase consisted of a phosphate buffer and methanol in a ratio of 100:900 (v/v). The flow rate was set at 1.0 mL/min, with an injection volume of 10 µL and a run time of 6 min using isocratic elution. UV detection was performed at a wavelength of 313 nm, and the temperature was maintained at 30°C. The analysis showed well-separated peaks with a high number of theoretical plates, a low tailing factor, and consistent retention time. Validation of the method was conducted, and all validation parameters were found to be within the acceptable limits. A method that is simple, accurate, and precise has been developed to estimate the amount of tapinarof in a topical formulation and active pharmaceutical ingredient. The optimized method involved the use of a column temperature set at 30°C, 90% methanol as the mobile phase, and a flow rate of 1.0 mL/min. The retention time for tapinarof was determined to be 2.88 min. The method exhibited linearity in the concentration range of 5 to 30 µg/mL (with an R2 value greater than 0.999) for tapinarof. The topical formulated cream and active pharmaceutical ingredient showed more than 90% dissolution within 5 min. The method developed in this study utilized photo diode array (PDA) for peak integrity and purity confirmation, making it suitable for the quantification of tapinarof in both topical formulations and active pharmaceutical ingredients. The method was validated and can be recommended for routine analysis in QC laboratories.

Preparation of polyacrylamide hydrophilic stationary phases with adjustable performance

Polymer modified silica materials are widely used as stationary phases in hydrophilic interaction liquid chromatography (HILIC), whereas a stationary phase with excellent performance is highly desired. In this study, vinyl modified silica was first synthesized through a silane coupling reaction, and then a polyacrylamide modified silica (PAM-SIL) stationary phase was successfully prepared using acrylamide as a copolymer monomer via free radical polymerization. The retention behaviors of polar analytes on the stationary phase under various chromatographic conditions, including acetonitrile content, buffer concentration and pH values were investigated, and a typical hydrophilic interaction retention mechanism was inferred. Exceptionally, the separation performance of the stationary phases could be regulated by controlling the polymer structure. Model analytes separated rapidly on the stationary phase which has an optimal grafting amount of vinyl, with the highest number of theoretical plates of orotic acid reaching 119,966/m. While the stationary phases with high acrylamide concentrations exhibited enhanced retention behavior and higher resolution for analytes. The adjustable separation performance will have huge potential in future separation and analysis applications.

Homomesoporous Metal-Organic Framework for High-Performance Electrochromatographic Separation.

Metal-organic frameworks (MOFs) have exhibited tremendous potential in the area of separation science. However, most of the developed MOF-based stationary phases contained only microporous structures and suffer from limited separation performance. Herein, homomesoporous MOFs with excellent mass transfer capability and strong thermodynamic interactions are first explored as the novel stationary phase for high-performance capillary electrochromatographic separations. As a proof of concept, noninterpenetrated mesoMOF-1 with uniform mesopore sizes (22.5 × 26.1 Å) and good stability was facilely grown on the inner surface of capillaries and applied as a homomesoporous MOF coating-based stationary phase for high-efficiency electrochromatographic separation. Seven types of analytes with different molecular dimensions were all baseline separated on a mesoMOF-1 coated column with high theoretical plate numbers and excellent repeatability, exhibiting significantly improved separation selectivity and column efficiency in comparison to a microporous HKUST-1 coated column. The maximum column efficiencies of the mesoMOF-1 coated column for substituted benzenes and halobenzenes reached up to 1.4 × 105 plates/m, and its mass loadability was also much higher than that of the HKUST-1 coated column. In addition, based on the analysis of adsorption kinetics and chromatographic retention behaviors, the interaction and retention mechanisms of different molecular-weight analytes on mesoMOF-1 coated stationary phases were systematically explored and disclosed in detail. These results indicate that the homomesoporous MOF-based stationary phase can effectively balance the kinetic diffusion (mass transfer capability) and thermodynamic interactions (the strength of adsorption interaction), having great potential for high-performance chromatographic separation.

Study of the separation ability differences of three covalent organic frameworks as coated materials in capillary electrochromatography

Covalent organic frameworks (COFs) have great potential applications in chromatographic separation. So it is crucial to understand the relationship between the separation ability of COFs and their structures. Here we report a strategy to evaluate the separation ability of three 2D COFs and explore the relationship between separation ability and their molecular structures. The three 2D COFs (COF-LZU1, COF-42 and COF-LZU8) have one same building unit 1,3,5-triformylbenzene, while varied from the conjugated linking units and functional side-chains. They were used to construct coated capillary column for capillary electrochromatographic separation of same groups of phthalates. They exhibited different separation efficiencies. COF-42 and COF-LZU8 coated capillary columns provided good signal resolutions and high column efficiencies with high theoretical plate numbers. It is demonstrated that COFs with hydrazone unit and longer side-chains provided higher selectivity and resolutions for the phthalates separation. Molecular simulations and DFT calculations were further proceeded to explore the deep reason why the three COFs coated CEC displayed different separation ability based on the host−guest interactions on molecular level. This work highlights a new opportunity to select or design functional COFs and improve their efficiency in chromatographic separation based on host−guest chemistry.

Characterization of linearly coupled capillaries with various inner diameters in the context of capillary electrophoresis

As a result of continuous instrumental progress, capillary electrophoresis has become an established separation technique. However, the choice of the suitable capillary inner diameter is sometimes difficult due to different instrumental requirements concerning injection, separation, or detection. To overcome this problem, we assembled two capillaries with different inner diameters, meaning that the inner diameter of the capillary at the injection side was different from that at the detection side. Since this was a rather uncommon approach, we focused on the associated effects in this proof-of-concept study. For the experiments, a non-aqueous model system was used, consisting of an acetonitrile-based background electrolyte and the two ferrocene derivates, ferrocenemethanol and decamethylferrocene. Using capillary flow injection analysis hyphenated to capacitively coupled contactless conductivity detection, it could be shown that fragmented capillaries of the same inner diameter had slightly lower volume flow rates than non-fragmented capillaries. With non-aqueous capillary electrophoresis hyphenated to UV detection, it was found that the coupling of capillaries with different inner diameter had a much stronger effect on the capillary electrophoresis flow than combinations with the same inner diameter. Additionally, if the inner diameter of the second capillary was larger than the inner diameter of the first capillary, a higher theoretical plate number and an increased sensitivity were found. Furthermore, it was found that there was no significant peak tailing introduced by the coupling.Graphic abstract

Non-aqueous electrophoresis integrated with electrospray ionization mass spectrometry on a thiol-ene polymer-based microchip device.

Non-aqueous capillary electrophoresis (NACE) on microfluidic chips is still a comparatively little explored area, despite the inherent advantages of this technique and its application potential for, in particular, lipophilic compounds. A main reason is probably the fact that implementation of NACE on microchips largely precluded the use of polymeric substrate materials. Here, we report non-aqueous electrophoresis on a thiol-ene-based microfluidic chip coupled to mass spectrometry via an on-chip ESI interface. Microchips with an integrated ESI emitter were fabricated using a double-molding approach. The durability of thiol-ene, when exposed to different organic solvents, was investigated with respect to swelling and decomposition of the polymer. Thiol-ene exhibited good stability against organic solvents such as methanol, ethanol, N-methylformamide, and formamide, which allows for a wide range of background electrolyte compositions. The integrated ESI emitter provided a stable spray with RSD% of the ESI signal ≤8%. Separation efficiency of the developed microchip electrophoresis system in different non-aqueous buffer solutions was tested with a mixture of several drugs of abuse. Ethanol- and methanol-based buffers provided comparable high theoretical plate numbers (≈ 6.6 × 104-1.6 × 105m-1) with ethanol exhibiting the best separation efficiency. Direct coupling of non-aqueous electrophoresis to mass spectrometry allowed for fast analysis of hydrophobic compounds in the range of 0.1-5μgmL-1 and 0.2-10μgmL-1 and very good sensitivities (LOD ≈ 0.06-0.28μgmL-1; LOQ ≈ 0.20-0.90μgmL-1). The novel combination of non-aqueous CE on a microfluidic thiol-ene device and ESI-MS provides a mass-producible and highly versatile system for the analysis of, in particular, lipophilic compounds in a wide range of organic solvents. This offers promising potential for future applications in forensic, clinical, and environmental analysis. Graphical abstract.

Increasing the Efficiency and Selectivity of Separation of Components in Chromatographic Analysis of Natural Gas

The composition of natural gas plays an important role in gas processing and transportation, and the main method for determination of the composition of natural gas is gas chromatography. In gas chromatography the efficiency of separation and the selectivity are the main characteristics of the analytical process. In this work the influence of the type of packed column on separation efficiency and selectivity during chromatographic analysis of natural gas under various conditions was investigated. The highest theoretical plate number was obtained with a Porapak R 4233 column (3 m), the best symmetry of the peaks was obtained with an OPN/Porous-Sil C Column 4355 (3 m), and the highest resolution was obtained with the Hayesep Q# 1754 (3 m), Hayesep D 4741 (2 m), and Porapak Q 1723 (2 m) columns. It was shown that optimization of operation conditions can significantly improve the efficiency and selectivity of chromatographic determination of the composition of natural gas.

Development and Validation of Simple, Rapid and Sensitive High- Performance Liquid Chromatographic Method for the Determination of Butenafine Hydrochloride

Aims: The current paper reports a simple, rapid, sensitive, accurate, and precise Reverse-phase high performance liquid chromatography (RP-HPLC) method with wide range of estimation to determine butenafine hydrochloride in nanosponges. This method has been validated as per ICH norms.&#x0D; Study Design: Experimental design with influence of variables such as mobile phase composition, flow rate, temperature and wavelength on the chromatographic peaks.&#x0D; Place and Duration of Study: Department of Pharmaceutics, College of Pharmacy, Prince Sattam Bin Abdulaziz University, Al-Kharj, Saudi Arabia between Jan 2020 and March 2020.&#x0D; Methodology: Separation was achieved by utilizing the most commonly used reverse phase column (C-18, 5 μm, 150 mm x 4.6 mm) set at 30ºC and quantified by UV detection at 280 nm after isocratic elution from a mobile phase (70:30 v/v of methanol: phosphate buffer pH 3.0) flowing at 1 ml/min.&#x0D; Results: A sharp and symmetrical peak was observed at 4.08 ± 0.01 minutes. The low variation in peak area and retention time (1.12% and 0.29%, respectively) and a high number of theoretical plates (&gt;2000) indicated this method’s efficiency and suitability. The least square linear regression analysis (Y = 9265.5 X + 1961.4) showed excellent correlation (r2 = 0.999 ± 0.0003) between concentration and peak area of butenafine hydrochloride through a wide concentration range of 1–50 µg/ml. The limits of detection and quantification (LOD and LOQ) were 0.18 µg/ml and 0.57 µg/ml, respectively. The assay or determinations were accurate, precise and reproducible with mean accuracy and mean relative standard deviation of precision of 101.53 ± 0.43% and 0.51 ± 0.11% respectively.&#x0D; Conclusion: The developed RP-HPLC method was simple, sensitive, reproducible with wide range of estimation of butenafine hydrochloride in the nanosponges. The proposed method could be used for the analysis of butenafine hydrochloride in the conventional pharmaceutical formulations such as tablets, syrup, creams including novel formulations such as nanoparticles, nanosponges, nanoemulsions. The proposed method overcomes the specificity, sensitivity and reproducibility related issues of ultraviolet-visible spectroscopy.

Microfluidic fast chiral separation of baclofen and phenylalanine enantiomers based on cyclodextrin-electrokinetic chromatography

A green chiral lab-on-a-chip cyclodextrin-based microchip electrophoretic method was developed for enantioseparation of baclofen (BCN) and phenylalanine (Phe) enantiomers for the first time. All BCN and Phe enantiomers were offline labeled with 4-fluoro-7-nitrobenzofurazan (NBD-F) or fluorescein-5-isothiocyanate (FITC) fluorogenic reagents prior to microfluidic injection in a dynamically coated polymethylmethacrylate microchip. The enantioseparation was performed using borate buffer solution (50 mM; pH 9.5) containing a dynamic coating polymer (methylcellulose) and a chiral selector of cyclodextrin (CD) family as the background electrolyte. The combination of sieving properties of methylcellulose polymer and inclusion complex formation properties of the cyclodextrins rendered a novel green pseudo-stationary phase for the microfluidic chiral separation of the studied enantiomers in short separation time. Seven CD chiral selectors were investigated in our study to illustrate the proposed enantioseparation mechanism in the dynamically coated microfluidic channels. The results indicated that use of a modified cyclodextrin, heptakis-(2,6-di-O-methyl)-β-cyclodextrin (HDM-β-CD), resulted in extra interactions with the analytes of interest, leading to extra resolution with high number of theoretical plates. Therefore, HDM-β-CD was able to separate the NBD-labeled and FITC-labeled enantiomers of BCN and Phe within short analytical time. Sensitivity problem of microchip electrophoresis was overcome by application of field-enhanced injection stacking which improved sensitivity to a higher extent by online preconcentration of the labeled enantiomers in the microfluidic channels. The method was fully validated and successfully applied for the assay of BCN enantiomers and Phe enantiomers in their pure racemic mixtures. Moreover, the developed method was applied for precise analysis of BCN enantiomers in pharmaceutical formulations and in biological samples. The extraction recovery values of BCN enantiomers in biological samples were > 87.5% and %RSD < 7.0.

Determination of three antiepileptic drugs in pharmaceutical formulations using microfluidic chips coupled with light-emitting diode induced fluorescence detection.

In this study, a facile, sensitive, and precise lab-on-a-chip electrophoretic method coupled with light-emitting diode induced fluorescence (LED-IF) detection was developed to assay three antiepileptic drugs, namely, vigabatrin, pregabalin, and gabapentin, in pharmaceutical formulations. The analytes were derivatised offline for the first time with fluorescine-5-isothiocyanate (FITC) to yield highly fluorescent derivatives with λex/em of 490/520nm. The FITC-labelled analytes were injected, separated, and quantitated by a microfluidic electrophoresis device using fluorescence detection. The labelled analytes were monitored using a blue LED-IF system. The separation conditions were significantly optimised adding specific concentrations of heptakis-(2,6-di-O-methyl)-β-cyclodextrin (HDM-β-CD) and methylcellulose to the buffer solution (40mM borate buffer). HDM-β-CD acted as a selective host for the studied antiepileptic drugs, rendering a high separation efficiency. Methylcellulose was used as an efficient dynamic coating polymer to prevent the labelled drugs from being adsorbed on the inner surfaces of the poly (methylmethacrylate) microchannels. A laboratory-prepared ternary mixture of the three antiepileptic drugs was separated within 100s with acceptable resolution between all the three analytes (Rs>3) and a high number of theoretical plates (N) for each analyte (N≈106 plates/m). The sensitivity of the method was enhanced approximately 80-fold by stacking to yield a detection limit below 0.6ngmL-1 in the concentration range of 2.0-200.0ngmL-1. The method was successfully validated for analysing the studied drugs in their pharmaceutical formulations.

Development and Validation of HPLC Method for DAAF and its Applications in Quality Control and Environmental Monitoring

Abstract3,3‐Diamino‐4,4‐azoxyfurazan (DAAF) as a promising energetic material, has many potential applications in both civil and military areas. High‐performance liquid chromatography (HPLC) is an important analytical method for quality control and environmental monitoring of DAAF. In this study, chromatographic conditions of DAAF were optimized to develop an analytical method based on HPLC‐DAD. DAAF and its byproducts were separated within 13 min with DAAF's retention time of 8.690 min and the resolution of all peaks were above 1.5 with high number of theoretical plates (N&gt;54259) under optimum condition of HPLC. Meanwhile, the method was validated and showed good performance for quantitative analysis of DAAF with LOD of 0.19 ng and linearity range from 0.84 to 211.16 μg mL−1. Furthermore, the method was applied to the analysis of the purity of DAAF for quality control of different preparation stages and trace DAAF in wastewater for environmental monitoring around explosive manufacturing.

Chiral separation and determination of amino acid enantiomers in fruit juice by open-tubular nano liquid chromatography.

A novel chiral porous-layer stationary phase was developed for use in open-tubular nano liquid chromatography. The stationary phase was prepared by an in-situ polymerization of 3-chloro-2-hydroxypropylmethacrylate (HPMA-Cl) and ethylene dimethacrylate (EDMA). The reactive chloro groups at the surface of the porous stationary phase were reacted with β-Cyclodextrin (β-CD). The resulting morphology was characterized by using scanning electron microscopy (SEM) and Fourier-transform infrared spectroscopy (FT-IR). The chromatographic performance of the column was evaluated by hydrophilic interaction chromatography (HILIC). Amino acids were used as test solutes. The running buffer conditions for the enantioseparation were found to be 85% acetonitrile (ACN):10%MeOH: 5% H2 O at 0.1% v/v trifluoro acetic acid (TFA) (flow rate: 800nL/min). The enantioseparation provided high theoretical plate numbers up to 26000 platesm-1 . A good retention capacity within satisfactory retention times was also achieved. Real sample applicability of this column to the separation of amino acid enantiomers in fruit juice sample was demonstrated.

Concentrical coils counter-current chromatography for natural products isolation: Salvia miltiorrhiza Bunge as example

Countercurrent chromatography (CCC) is an efficient separation technique without the solid support matrix, largely depending on the partition of two-immiscible liquid phases in the separation column. Since the helical coil planet centrifuge was invented in early 1970s by Yoichiro Ito, a series of coils columns, including spiral coils and conical coils columns have been developed for CCC separation. In this work, we introduced a new simple and efficient concentrical coils column for CCC separation, which was prepared by winding the whole polytetrafluoroethylene (PTFE) tube into the circular grooves from the rotation axis in the same direction. Once the PTFE tube filled in all space of one round of the circular groove, it was jumped into the nearby outer circular groove through the gap and until the whole groove was filled. The three same concentrical coils distributed on three disc-shaped holders were connected by the same PTFE tube to form concentrical coils separation column. The separation capacity was further investigated using ten tanshinones of the extracts of a Traditional Chinese Medicine Salvia miltiorrhiza Bunge as a model natural product. All results indicated that the concentrical coils column could hold satisfactory retention of the stationary phase, higher theoretical plate number and better resolution for CCC separation of more than ten tanshinones. It may be an alternative CCC column for non-targeted and targeted isolation of bioactive natural products.

Rationale behind the optimum efficiency of columns packed with new 1.9 μm fully porous particles of narrow particle size distribution

Columns packed with new commercially available 1.9 fully porous particles of narrow particle size distribution (nPSD) are characterized by extremely high efficiency. Under typical reversed phase conditions, these columns are able to generate very high number of theoretical plates (in the order of 300,000plates/m and more). In this paper, we investigate the origin of the high performance of these nPSD columns by performing a series of measurements that include, in addition to the traditional determination of the van Deemter curve, peak parking, pore blocking and inverse size exclusion experiments. Two nPSD columns (both 100×3.0mm) have been considered in this study: the first one, packed with particles of 80Å pore size, is commercially available. The second one is a prototype column packed with 1.9 fully porous particles of 120Å pore size.The main conclusion of our study is that these nPSD columns are characterized by extremely low eddy dispersion, while longitudinal diffusion and mass transfer kinetics are substantially equivalent to those of other fully porous particles of similar chemistry.

Comprehensive untargeted lipidomic analysis using core–shell C30 particle column and high field orbitrap mass spectrometer

The goal of untargeted lipidomics is to have high throughput, yet comprehensive and unambiguous identification and quantification of lipids. Novel stationary phases in LC separation and new mass spectrometric instruments capable of high mass resolving power and faster scanning rate are essential to achieving this goal. In this work, 4 reversed phase LC columns coupled with a high field quadrupole orbitrap mass spectrometer (Q Exactive HF) were thoroughly compared using complex lipid standard mixture and rat plasma and liver samples. A good separation of all lipids was achieved in 24min of gradient. The columns compared include C30 and C18 functionalization on either core–shell or totally porous silica particles, with size ranging from 1.7 to 2.6μm. Accucore C30 column showed the narrowest peaks and highest theoretical plate number, and excellent peak capacity and retention time reproducibility (<1% standard deviation). As a result, it resulted in 430 lipid species identified from rat plasma and rat liver samples with highest confidence. The high resolution offered by the up-front RPLC allowed discrimination of cis/trans isomeric lipid species, and the high field orbitrap mass spectrometer afforded the clear distinction of isobaric lipid species in full scan MS and the unambiguous assignment of sn-positional isomers for lysophospholipids in MS/MS. Taken together, the high efficiency LC separation and high mass resolving MS analysis are very promising tools for untargeted lipidomics analysis.

Preparation of a novel ionic hybrid stationary phase by non-covalent functionalization of single-walled carbon nanotubes with amino-derivatized silica gel for fast HPLC separation of aromatic compounds

Single-walled carbon nanotubes (SWCNTs) were immobilized on spherical silica gel with a 4-μm average particle size and a 60-Å average pore size. The amino-derivatized silica gel was non-covalently coated with carboxylated SWCNTs to preserve the structure of the nanotubes and their physico–chemical properties. The novel ionic hybrid stationary phase was characterized by scanning electron microscopy (SEM), infra-red (IR) spectroscopy and elemental analysis, and then, it was used to fill an empty 150×4.6mm2 high-performance liquid chromatography (HPLC) column. Chromatographic parameters, such as the theoretical plate number, retention factor and peak asymmetry factor, and analytical parameters, such as the limit of detection (LOD), limit of quantification (LOQ), linear range, calibration equation, and R2 value, and quantitative analysis parameters were calculated for all of the analytes. Using different mobile phases, five different classes of aromatic hydrocarbons were separated in a very short analysis time of 4–8min. Furthermore, a high theoretical plate number (up to 25000) and an excellent peak asymmetry factor (1.0) were obtained. The results showed that the surface of the SWNTs had very strong interactions with aromatic groups, therefore providing high selectivity for the separation of different classes of aromatic compounds. This study indicates that SWCNTs enable the extension of the application range of the newly prepared stationary phases for the fast separation of aromatic compounds by HPLC.

Expanding the elution by characteristic point method to columns with a finite number of theoretical plates

The elution by characteristic point (ECP) method provides a rapid approach to determine whole isotherm data with small material usage. It is especially desired wherever the adsorbent or the adsorbate is expensive, toxic or only available in small amounts. However, the ECP method is limited to adsorbents that are well optimized for chromatographic use and therefore provide a high number of theoretical plates when packed into columns (2000 or more for Langmuir type isotherms are suggested). Here we present a novel approach that uses a new profile correction to apply the ECP method to poorly optimized adsorbents with less than 200 theoretical plates. Non-ideality effects are determined using a dead volume marker injection and the resulting marker profile is used to compensate the named effects considering their dependency from the actual concentration instead of assuming rectangular profiles. Experimental and literature data are used to compare the new ECP approach with batch method results.

Tunable thick porous silica coating fabricated by multilayer-by-multilayer bonding of silica nanoparticles for open-tubular capillary chromatographic separation

A simple coating procedure employing a multilayer-by-multilayer process to modify the inner surface of bare fused-silica capillaries with silica nanoparticles was established. The silica nanoparticles were adsorbed onto the capillary wall via a strong electrostatic interaction between amino functional groups and silica particles. The thickness of the coating could be tuned from 130 to 600nm by increasing the coating cycles from one to three. Both the retention factor and the resolution were greatly increased with increasing coating cycles. The loading capacity determined by naphthalene in the column with three coating cycles is 152.1pmol. The effects of buffer concentration and pH value on the stability of the coating were evaluated. The retention reproducibility of the separation of toluene was 0.8, 1.2, 2.3, and 4.5%, respectively, for run-to-run, day-to-day, column-to-column, and batch-to-batch, respectively. The chromatographic performance of these columns was evaluated by both capillary liquid chromatography and open-tubular capillary electrochromatography (OT-CEC). Separation of aromatic hydrocarbons in the column with three coating cycles provided high theoretical plate numbers (up to 269,280 platesm−1 for toluene) and short separation time (<15min) by using OT-CEC mode. The method was also used to separate egg white proteins. Both acidic and basic proteins as well as four glycoisoforms were separated in a single run.

New pressure-assisted sweeping on-line preconcentration for highly polar environmentally relevant nitrosamines: Part 2. Cationic and anionic surfactants with zero-flow capillaries.

We recently introduced a pressure-assisted sweeping-reversed migration-EKC (RM-EKC) method for preconcentration of neutral polar N-nitrosamines with low affinity for the micellar phase. The type of surfactant and phase ratio are dominant factors in dictating the magnitude of interactions between analyte and micellar phase, thus four surfactants (anionic and cationic) with a range of functionalities (SDS, ammonium perfluorooctanoate (APFO), bile salts, and cetyltrimethylammonium chloride (CTAC)) were evaluated for sweeping-RM-EKC of highly polar N-nitrosamines. All gave acceptable results for sweeping-RM-EKC when used in high concentrations (≥200 mM) with low EOF. While no single surfactant was superior by all measures, all but the bile salts had useful performance characteristics. APFO showed the narrowest peak widths and highest number of theoretical plates, though two species co-migrated at all concentrations (25-300 mM); SDS and the cationic surfactant CTAC also showed good separation characteristics and could resolve all peaks, but CTAC had wider separation window. Various types of capillaries coated for EOF control were compared for use with anionic and cationic surfactants. A commercial zero-EOF capillary coated with a polymer bearing sulfonic acid functional groups showed superior EOF suppression and reproducibility of migration time with all surfactants.