Search Results (16)

In this study, new monolithic poly(9-anthracenylmethyl methacrylate-co-trimethylolpropane trimethacrylate (TRIM) columns, referred as ANM monoliths were prepared, for the first time, and were used for the separation media for biomolecules and proteomics analysis by nano-liquid chromatography (nano-LC). Monolithic columns were prepared by in situ polymerization of 9-anthracenylmethyl methacrylate (ANM) and trimethylolpropane trimethacrylate (TRIM) in a fused silica capillary column of 100 µm ID. Polymerization solution was optimized in relation to monomer and porogenic solvent. Scanning electron microscopy (SEM) and chromatographic analyses were performed for the characterization studies of ANM monoliths. The ANM monolith produced more than 46.220 plates/m, and the chromatographic evaluation of the optimized ANM monolith was carried out using homologous alkylbenzenes (ABs) and polyaromatic hydrocarbons (PAHs), allowing both strong hydrophobic and π-π interactions. Run-to-run and column-to-column reproducibility values were found as <2.91% and 2.9–3.2%, respectively. The final monolith was used for biomolecule separation, including both three dipeptides, including Alanine-Tyrosine (Ala-Tyr), Glycine-Phenylalanine (Gly-Phe), and L-carnosine and five standard proteins, including ribonuclease A (RNase A), α-chymotrypsinogen (α-chym), lysozyme (Lys), cytochrome C (Cyt C), and myoglobin (Mb) in order to evaluate its potential. Both peptides and proteins were baseline separated using the developed ANM monolith in nano-LC. The ANM monolith was then applied to the protein and peptide profiling of MCF-7 cell line, which allowed a high-resolution analysis of peptides, providing a high peak capacity. Full article

This work is associated with the preparation of capillary chromatographic columns containing inorganic-organic composites comprised of naturally occurring montmorillonite (MMT) clay mineral and polymethacrylate monolithic material. The prepared composites combine the best qualities of both constituents, offering desirable properties for use under the disparate conditions of both GC and HPLC at the same time. The stationary phases were investigated by scanning electron microscopy (SEM), the specific surface area, and thermogravimetric analysis (TGA) and examined in terms of various conditions utilized for GC and HPLC methods. The prepared columns demonstrated an excellent permeability and stability against common chromatographic conditions, such as the eluent type, flow rate, pressure, and temperature. The results confirmed that the addition of small amounts of MMT into the monolith induced significant improvement in the specific surface area, which contributed to the formation of more active sites and enhanced the retention of analytes. The registered column backpressures did not exceed 980 kPa and 16,500 kPa for the prepared GC and HPLC columns, respectively. The prepared columns were subjected to the separation of various interesting compounds possessing different chemistries and polarities, including alkanes, alkylbenzenes, polycyclic aromatic hydrocarbons (PAHs), alcohols, ketones, phenols, some common organic solvents, and isomeric mixtures. Under the optimal conditions, the efficiency of the columns fell between 4900–38,500 plates m⁻¹ for GC and 3400–58,800 plates m⁻¹ for capillary HPLC applications. In all cases, the measured chromatographic resolution was more than 1.38, with excellent an peak symmetry and low tailing factors. In comparison with the most commonly used commercial columns, the polysiloxane open tubular column for GC and silica-based C₁₈ packed column for HPLC, the prepared GC columns demonstrated a faster separation with a higher efficiency, comparable resolution and tailing factors, and lower consumption of carrier gas. Regarding the capillary columns prepared for HPLC, the chromatographic experiments exposed a much lower run time with a comparable efficiency and resolution and drastically lower consumption of mobile phase solvents and samples. The results demonstrate that the MMT-based polymethacrylate monolith composites are applicable as novel and promising separation media for analyzing various mixtures of interest in different fields, such as petrochemical and environmental samples. Full article

Separation with high efficiency and good resolution is constantly in demand in the pharmaceutical industry. The fast and efficient separation of complex samples such as peptides and proteins is a challenging task. To achieve high efficiency with good resolution, chromatographers are moving towards small particles packed into narrow-bore columns. Silica monolith particles (sub-2 µm) were derivatized with chlorodimethyl octadecyl silane (C18) and packed into stainless steel columns (100 mm × 1.8 mm i.d) by a slurry-packing method. The developed columns were used for the separation of peptides and proteins. A separation efficiency (N) of 40,000 plates/column (400,000 plates/m) was achieved for the mixture of five peptides. Similarly, the fast separation of the peptides was carried out using a high flow rate, and the separation of the five peptides was achieved in one minute with high efficiency (N ≅ 240,000 plates/m). The limit of detection (DL) and the limit of quantification (QL) for each analyte were determined by developing a linear regression curve with relatively very low concentrations of the target compound. The average values of the QL for the peptide and proteins were 0.55 ng and 0.48 ng, respectively, using short C18 column (1.8 mm × 100 mm) UV (at 214 nm). The fast analysis of peptides and proteins with such high efficiency and good resolution has not been reported in the literature yet. Owing to high efficiency, these home-made columns could be used as an alternative to the expensive commercial columns for peptide and protein separation. Full article

Ribavirin, a nucleoside analog, is used to treat chronic hepatitis C (HCV) infections. Therapeutic drug monitoring for ribavirin is useful for predicting the effect of treatment. In this study, the selective extraction of ribavirin from serum samples and the HPLC-UV detection method were investigated using a monolithic silica disk-packed spin column with phenylboronate moieties. In this study, 0.6% ammonia and 1% formic acid solutions were used as the conditioning and elution solutions, respectively, and recoveries of >90% were obtained. Ribavirin was separated on an InertSustain AQ-C18 column by isocratic elution. The mobile phase consisted of a mixture of 7 mM Na₂SO₄ and 60 mM H₃PO₄ in H₂O. Linear regression curves were observed for calibrations over a concentration range of 0.25–25 µg/mL. The lower limit of detection was 0.05 µg/mL, and the lower limit of quantification was 0.1 µg/mL. The intra- and inter-day precisions were below 3.2 and 3.1%, respectively. This method can be applied to quantify ribavirin levels in human serum and may be useful for pharmacokinetic studies. Full article

A new feature of hydrophobic fumed silica nanoparticles (HFSNPs) when they apply to the preparation of monolithic nano-columns using narrow monolithic fused silica capillary columns (e.g., 50-µm inner diameter) was presented. The monolithic nano-columns were synthesized by an in-situ polymerization using butyl methacrylate (BMA) and ethylene dimethacrylate (EDMA) at various concentrations of AEROSIL^®R972, called HFSNPs. Dimethyl formamide (DMF) and water were used as the porogenic solvents. These columns (referred to as HFSNP monoliths) were successfully characterized by using scanning electron microscopy (SEM) and reversed-phase nano-LC using alkylbenzenes and polyaromatic hydrocarbons as solute probes. The reproducibility values based on run-to-run, column-to-column and batch-to-batch were found as 2.3%, 2.48% and 2.99% (n = 3), respectively. The optimized column also indicated promising hydrophobic interactions under reversed-phase conditions, while the feasibility of the column allowed high efficiency and high throughput nano-LC separations. The potential of the final HFSNP monolith in relation to intact protein separation was successfully demonstrated using six intact proteins, including ribonuclease A, cytochrome C, carbonic anhydrase isozyme II, lysozyme, myoglobin, and α-chymotrypsinogen A in nano-LC. The results showed that HFSNP-based monolithic nanocolumns are promising materials and are powerful tools for sensitive separations. Full article

Efficient separation of pharmaceuticals and metabolites with the adequate resolution is a key factor in choosing the most suitable chromatographic method. For quality control, the analysis time is a key factor, especially in pharmacokinetic studies. High back pressure is considered as one of the most important factors in chromatography’s flow control, especially in UHPLC. The separation of the anti-hyperlipidemic mixtures was carried out using two columns: a column silica-based particle packed UHPLC and a monolithic column. The systematic suitability of the two columns was compared for the separation of Fenofibrate, its active metabolite, Fenofibric acid and Pravastatin using Atorvastatin as an internal standard. Separation on both columns was obtained using ethanol: buffer potassium dihydrogen orthophosphate pH = 3 (adjusted with orthophosphoric acid) (75:25 v/v) as mobile phase and flow rate 0.8 mL/min. The analytes’ peak detection was achieved by using a PDA detector at 287 nm, 214 nm, 236 nm, and 250 nm for Fenofibrate, Fenofibric acid, Pravastatin, and Atorvastatin, respectively. Reduction of back-pressure was achieved with the monolithic column, where the analytes could be completely separated in less than 1.5 min at a flow rate of 5 mL/min. The principles of Green Analytical Chemistry (GAC) were followed throughout the developed method using environmentally safe solvents. Full article

Water pollution is a severe worldwide issue. Constructing advanced porous composite materials has been an efficient route to water remediation via adsorption. In this study, a unique microspheres-in-pores monolithic structure was fabricated. An emulsion-templated polymer monolith was first prepared and silica microspheres were subsequently formed in the porous polymer. A silica precursor was modified with a fluorescent dye and co-condensed with other precursors to fabricate porous composites with fluorescent properties, which were enhanced by the presence of Ag nanoparticles in the polymer matrix. This unique material showed good promise in water remediation by removing organic dyes and heavy metal ions from wastewater via a flowing filter or monolithic column separation. Full article

This review draws attention to the use of chiral monolithic silica HPLC columns for the enantiomeric separation and determination of chiral compounds. Properties and advantages of monolithic silica HPLC columns are also highlighted in comparison to conventional particle-packed, fused-core, and sub-2-µm HPLC columns. Nano-LC capillary monolithic silica columns as well as polymeric-based and hybrid-based monolithic columns are also demonstrated to show good enantioresolution abilities. Methods for introducing the chiral selector into the monolithic silica column in the form of mobile phase additive, by encapsulation and surface coating, or by covalent functionalization are described. The application of molecular modeling methods to elucidate the selector–selectand interaction is discussed. An application for enantiomeric impurity determination is also considered. Full article

Chondroitin sulfate A was covalently immobilized onto a monolithic silica epoxy column involving a Schiff base formation in the presence of ethylenediamine as a spacer and evaluated in terms of its selectivity in enantioseparation. The obtained column was utilized as a chiral stationary phase in enantioseparation of amlodipine and verapamil using a mobile phase consisting of 50 mM phosphate buffer pH 3.5 and UV detection. Sample dilution by organic solvents (preferably 25% v/v acetonitrile-aqueous solution) was applied to achieve baseline enantioresolution (Rs > 3.0) of the individual drug models within 7 min, an excellent linearity (R² = 0.999) and an interday repeatability of 1.1% to 1.8% RSD. The performance of the immobilized column for quantification of racemate in commercial tablets showed a recovery of 86–98% from tablet matrices. Computational modeling by molecular docking was employed to investigate the feasible complexes between enantiomers and the chiral selector. Full article

Monolithic fillings used in chromatography are of great interest among scientists since the first reports of their synthesis and use were published. In the 20 years since silica-based monolithic columns were introduced into the commercial market, numerous papers describing their chromatographical properties and utility in various branches of industry and scientific investigations were presented. This review is focused on possible applications of commercially available silica-based HPLC monolithic columns in the analysis of biological samples. Full article

Over the past years, a great effort has been devoted to the development of new sorbents that can be used to pack or to coat extractive capillaries for in-tube solid-phase microextraction (IT-SPME). Many of those efforts have been focused on the preparation of capillaries for miniaturized liquid chromatography (LC) due to the reduced availability of capillary columns with appropriate dimensions for this kind of system. Moreover, many of the extractive capillaries that have been used for IT-SPME so far are segments of open columns from the gas chromatography (GC) field, but the phase nature and dimensions are very limited. In particular, polar compounds barely interact with stationary GC phases. Capillary GC columns may also be unsuitable when highly selective extractions are needed. In this work, we provide an overview of the extractive capillaries that have been specifically developed for capillary LC (capLC) and nano LC (nanoLC) to enhance the overall performance of the IT-SPME, the chromatographic separation, and the detection. Different monolithic polymers, such as silica C₁₈ and C₈ polymers, molecularly imprinted polymers (MIPs), polymers functionalized with antibodies, and polymers reinforced with different types of carbon nanotubes, metal, and metal oxide nanoparticles (including magnetic nanoparticles), and restricted access materials (RAMs) will be presented and critically discussed. Full article

Isoquinoline alkaloids are the main group of secondary metabolites present in Chelidonium majus extracts, and they are still the object of interest of many researchers. Therefore, the development of methods for the investigation and separation of the alkaloids is still an important task. In this work, the application potential of a silica-based monolithic column for the separation of alkaloids was assessed. The influence of the organic modifier, temperature, salt concentration, and pH of the eluent on basic chromatographic parameters such as retention, resolution between neighboring peaks, chromatographic plate numbers, and peak asymmetry were investigated. Based on the obtained results, a gradient elution program was developed and used to separate and quantitatively determine the main alkaloids in a Chelidonium majus root extract. Full article

A strategy for the preparation of silica-based monolithic capillary columns (150 × 0.1 mm) with high selectivity to amino acids is presented. The zwitterionic columns were prepared by coating the silica monolith with [2-(methacryloyloxy)ethyl]-dimethyl-(3-sulfopropyl)-ammonium hydroxide via 3-(trimethoxysilyl)propyl methacrylate. The columns were evaluated under isocratic conditions in hydrophilic interaction liquid chromatography. The best separation of amino acids was obtained on the monolithic column prepared by a stepwise modification procedure where the modification step was repeated four times. The mixture of fifteen amino acids was separated within 13 min using the mobile phase consisting of 75% acetonitrile and 25% 5 mmol/L ammonium acetate at pH 4.5. Full article

Candida albicans is one of the major pathogens that cause the serious infectious condition known as candidiasis. C. albicans was investigated by proteome analysis to systematically examine its virulence factors and to promote the development of novel pharmaceuticals against candidiasis. Here, we review quantitative time-course proteomics data related to C. albicans adaptation to fetal bovine serum, which were obtained using a nano-liquid chromatography/tandem mass spectrometry system equipped with a long monolithic silica capillary column. It was revealed that C. albicans induced proteins involved in iron acquisition, detoxification of oxidative species, energy production, and pleiotropic stress tolerance. Native interactions of C. albicans with macrophages were also investigated with the same proteome-analysis system. Simultaneous analysis of C. albicans and macrophages without isolating individual living cells revealed an attractive strategy for studying the survival of C. albicans. Although those data were obtained by performing proteome analyses, the molecular physiology of C. albicans is discussed and trials related to pharmaceutical applications are also examined. Full article

Avanafil (AVA), one of the most effective drugs prescribed for erectile dysfunction, is a pyrimidine-derivative PDE5 inhibitor. In the current work, new LC methods were developed and validated for quantitative determination of avanafil and qualitative determination of its degradation products. The quantitative determination of avanafil was carried out using liquid chromatography with photodiode array detection (LC-DAD) and liquid chromatography-tandem mass spectrometry LC-MS/MS methods, and fully validated according to the ICH Q2 (R1) guideline, while qualitative determination was performed using a liquid chromatography mass spectrometry-ion trap-time of flight (LCMS-IT-TOF) instrument. The separation of avanafil and its degradation products was carried out using the same reversed-phase chromatographic conditions, in which a second-generation C₁₈-bonded monolithic silica column (Chromolith^® High Resolution RP-18e, 100 × 4.6 mm, Merck KGaA) was used as stationary phase. Briefly, the methods enable quantitation of avanafil with high accuracy (recovery > 95%) and precision (RSD% < 2.0), within the ranges of 0.5–20 μg/mL for LC-DAD and 150–6000 ng/mL for LC-MS/MS. In the forced degradation studies, over and above currently existing data, a new oxidation-based degradation product, whose predicted m/z is 367.1168, was identified and its structure was confirmed by high-resolution mass spectrometric analysis. As the main advantage, either an LC-DAD or LC-MS/MS instrument can be chosen for interference-free quantitation of AVA, according to the facilities in quality-control laboratories. Full article