Volume 10, Issue 3 - Special Issue: Recent Advances in Separation Science

High-speed countercurrent chromatography (HSCCC) is a liquid-liquid separation chromatographic technique which uses no solid supporting medium. During its rapid development in the last 30 years, great progress has been made in the instrumentation, the basic study and application of HSCCC. It has significant advantages over other instrumental separation techniques, in its high efficiency and continuous processing capability. In recent years, HSCCC has been widely used in research and development of natural medicines and functional foods, including preparative separation, fingerprint analysis and screening of bioactive constituents. A review of the technique is presented in this paper.

The miniaturization and improvement of sample preparation is a challenge that has been fulfilled up to a point in many fields of analytical chemistry. Particularly, the hyphenation of microextraction with advanced analytical techniques has allowed the monitoring of target analytes in a vast variety of environmental samples. Several benefits can be obtained when miniaturized techniques such as solid-phase microextraction (SPME) or liquid-phase microextraction (LPME) are applied, specifically, their easiness, rapidity and capability to separate and pre-concentrate target analytes with a negligible consumption of organic solvents. In spite of the great acceptance that these green sample preparation techniques have in environmental research, their full implementation has not been achieved or even attempted in some relevant environmental matrices. In this work, a critical review of the applications of LPME and SPME techniques to isolate and pre-concentrate traces of organic pollutants is provided. In addition, the influence of the environmental matrix on the effectiveness of LPME and SPME for isolating the target organic pollutants is addressed. Finally, unsolved issues that may hinder the application of these techniques for the extraction of dissolved organic matter from environmental samples and some suggestions for developing novel and less selective enrichment and isolation procedures for natural organic matter on the basis of SPME and LPME are included.

Enantiomers (stereoisomers) can exhibit substantially different properties if present in chiral environments. Since chirality is a basic property of nature, the different behaviors of the individual enantiomers must be carefully studied and properly treated. Therefore, enantioselective separations are a very important part of separation science. To achieve the separation of enantiomers, an enantioselective environment must be created by the addition of a chiral selector to the separation system. Many chiral selectors have been designed and used in various fields, such as the analyses of drugs, food constituents and agrochemicals. The most popular have become the chiral selectors and/or chiral stationary phases that are of general use, i.e., are applicable in various separation systems and allow for chiral separation of structurally different compounds. This review covers the most important chiral selectors / chiral stationary phases described and applied in high performance liquid chromatography and capillary electrophoresis during the period of the last three years (2008–2011).

This review summarizes the development of capillary ion chromatography (CIC) over approximately the last 5 years. It mainly focuses on the technologic aspects of several key components associated with CIC, including micropump, microscale electrodialytic eluent generator, microscale suppressor and the detector.

Spreading sewage sludge on agricultural lands has been actively promoted by national authorities as an economic way of recycling. However, as by-product of wastewater treatment, sewage sludge may contain toxic substances, which could be incorporated into agricultural products or be distributed in the environment. Moreover, sediments can be contaminated by the discharge of wastewater effluents into rivers. This article reviews the determination of emerging contaminants (surfactants, flame retardants, pharmaceuticals and personal care products) in environmental solid samples (sludge, soil and sediment). Sample preparation, including extraction and clean-up, as well as the subsequent instrumental determination of contaminants are discussed. Recent applications of extraction techniques, such as Soxhlet extraction, ultrasound assisted extraction, pressurised liquid extraction, microwave assisted extraction and matrix solid-phase dispersion to the analysis of emerging contaminants in environmental solid samples are reviewed. Determination of these contaminants, generally carried out by gas chromatography and liquid chromatography coupled with different detectors, especially mass spectrometry for the identification and quantification of residues, is also summarised and discussed.

Booster biocides are organic compounds that are added to antifouling copper-based paints to improve their efficacy. Due to their widespread use, they are common pollutants of marine ecosystems. Some of these compounds show acute and chronic toxic effects in non-targeted organisms at concentrations as low as ng L−1. The determination of these compounds is therefore important, and for some, which are prioritized in the EU water framework directive, a necessity. Because of their low concentrations and the matrix effect, these contaminants often require a suitable sample preparation step (extraction/pre-concentration) prior to chromatographic determination. The aim of the present article is to review extraction and chromatographic methodologies related to the determination of common booster biocides in marine samples published in the scientific literature. These methodologies include liquid-liquid extraction (LLE), solid phase extraction (SPE), solid phase microextraction (SPME), single drop microextraction (SDME), Soxhlet extraction, microwave-assisted extraction (MAE), supercritical fluid extraction (SFE) and pressurized liquid extraction (PLE) as extraction methods, and both gas and liquid chromatography as determination techniques.

Hydrophilic interaction chromatography (HILIC) is an increasingly popular alternative to conventional HPLC for drug analysis. It offers increased selectivity and sensitivity, and improved efficiency when quantifying drugs and related compounds in complex matrices such as biological and environmental samples, pharmaceutical formulations, food, and animal feed. In this review we summarize HILIC methods recently developed for drug analysis (2006–2011). In addition, a list of important applications is provided, including experimental conditions and a brief summary of results. The references provide a comprehensive overview of current HILIC applications in drug analysis.

This review highlights the fundamentals and the most prominent advances in the field of HPLC instrumentation over the last decades. Fundamental aspects and practical considerations of column switching, conventional (heart-cut) and comprehensive two-dimensional LC are presented. Different aspects of microcolumn- and nanoliquid-chromatography are reviewed. Recent progress in column technology and the demands and developments in instrumentation and accessories for miniaturized LC are also discussed. In the field of miniaturization, particularly in chip-based nano-LC systems, some aspects on micro-fluidic chip fabrication, using particle-packed HPLC microchips or polymer-based monoliths, are addressed. An introduction to ultra performance LC (UPLC) is also presented.

Micellar liquid chromatography (MLC) is an analytical technique belonging to the wide range of reversed-phase liquid chromatographic (RP-LC) separation techniques. MLC with the use of surfactant solutions above its critical micellar concentration (CMC) and the addition of organic modifiers is currently an important analytical tool with still growing theoretical considerations and practical applications in pharmaceutical analysis of drugs and other biologically active compounds. The use of MLC as an alternative, relatively much faster in comparison to conventional chromatographic separation techniques has several advantages, especially as being suitable for screening pharmaceutical analysis. The analytical data received from MLC analysis are considered a useful source of information to predict passive drug absorption, drug transport and other pharmacokinetics and physicochemical measures of pharmaceutical substances. In the review several MLC assays for determination of drugs and other active compounds in biological samples were compared and critically discussed. The presented overview provides information on recent applications and achievements connected with the practical use of MLC. The review covers fields of interest related to theory and mechanism of MLC separation, direct applications of MLC in pharmaceutical analysis, including optimization and efficiency of separation with the use of modification of stationary phase and mobile phase compositions as well as the determination of physicochemical characteristics of drugs by MLC.

This article examines the contribution of liquid chromatography to the study of polyolefin additives commonly used to obtain improved environmental resistance (antioxidants, ultraviolet light stabilizers, antistatics, and so on) and appearance enhancements (e.g. colorants). Several reversed-phase liquid chromatographic methods are summarized, and a detailed description of different detectors is provided. In addition, ways of applying these methods to analyse food contact materials and plastic toys are emphasized. Finally, the potential use of these methods is addressed which complies with European health safety regulations.

Capillary electrophoresis (CE) has become one of the most useful tools in separation science because of its high separation efficiency, low cost, versatility, ease of sample preparation and automation. However, some limitations of CE, such as poor concentration sensitivity due to its lower sample loading and shorter optical path length, limits its further applications in separation science. In order to solve this problem, various on-line sample preconcentration techniques such as transient isotachophoresis preconcentration, field-enhanced sample stacking, micelle to solvent stacking, micelle collapse, dynamic pH junction, sweeping, solid phase extraction, single drop microextraction and liquid phase microextraction have been combined with CE. Recent developments, applications and some variants together with different combinations of these techniques integrating in CE are reviewed here and our discussions will be confined to the past three years (2008–2011).

Capillary electrophoresis (CE) is an attractive technique in separation science because of its high separation performance, short analysis time and low cost. Electrochemical detection (EC) is a powerful tool for CE because of its high sensitivity. In this review, developments of CE-EC from 2008 to August, 2011 are reviewed. We choose papers of innovative and novel results to demonstrate the newest and most important progress in CE-EC.

The tremendous potential of room temperature ionic liquids as an alternative to environmentally harmful ordinary organic solvents is well recognized. Due to their unique properties, such as low volatility, tunable viscosity and miscibility, and electrolytic conductivity, ionic liquids have attracted extensive attention and gained popularity in many areas of analytical chemistry including modern sample preparation techniques. In this review the advantages and limitations of application of ionic liquids as solvents/sorbents for microextraction are critically discussed. Topics covered include solid-phase microextraction, single drop microextraction, dispersive liquid-liquid microextraction and hollow-fiber liquid-phase microextraction. The compatibility of the ionic liquid-based microextraction with different analytical techniques such as gas chromatography, high-performance liquid chromatography, electrothermal or flame atomic absorption spectrometry and some others is also discussed. Finally, the main practical applications on this topic are summarized.

Carcinogenic and mutagenic properties of aflatoxin species are known in literature. Their intake over a long time period might be health-dangerous for human even at trace levels. It is well known that different foodstuffs can be contaminated by aflatoxin species through growing and storage. Due to the serious health effects, sensitive determination of aflatoxin species in any matrices related with the human being is very crucial at trace levels. In literature, there are sensitive techniques to analyze the different samples for the contents of their aflatoxin species. Each technique has some advantages and disadvantages over the other techniques. This review aims to summarize the different health effects of aflatoxin species, development of analytical techniques and applications of developed techniques in a variety of matrices.

Over the last years, interest in intranasal administration as an alternative and promising route for the delivery of drugs withlocal, systemic, and even central nervous system action has tremendously increased. Accordingly, understanding of the propertiesand characteristics of the nasal cavity as well as the biodisposition processes of drugs into the nasal compartments is acquiringa significant prominence in the field of pharmacology. In this context, the development and validation of bioanalytical methodologies for the quantitative measurement of drugs and their metabolites in nasal and paranasal tissues and/or secretions is of the utmostimportance. However, currently, information concerning bioanalysis of drugs in nasal and paranasal tissues and/or secretionsis scattered. This review aims to provide a valuable overview of the methodologies that have been used for the collectionand preparation of nasal and paranasal samples with special emphasis placed on the review of liquid chromatographic methodsemployed for the quantitative determination of small-molecule drugs and their metabolites in such specimens.

Direct Analysis in Real Time mass spectrometry (DART-MS) is an emerging and rapidly developing area of ambient desorption ionization mass spectrometric techniques. Its coupling with planar chromatography is especially promising, as compared to other ambient desorption ionization techniques, because it does not require the use of liquids that may distort the shape of a spot by diffusion effects. In the first publications on TLC/HPTLC-DART-MS, due to the fixed, horizontally aligned supply of the gas flow from the DART ionization source to the MS inlet, the introduction of HPTLC/TLC plates as cut strips was inconvenient for quantitation, and the repeatability was very low due to the manual positioning. Recently a new version of the DART ion source was suggested, which allows adjusting the angle of the DART gas stream and the use of a motorized rail, thereby, improving highly the capabilities of TLC/HPTLC-DART-MS. This comprehensive review describes the development and analytical capabilities of TLC/HPTLC-DART-MS, and the general DART-MS perspectives for surface analysis or imaging MS.

Non-steroidal anti-inflammatory drugs, as a therapeutic class, are among the most often used active pharmaceutical ingredients in heath care in the world. They are mostly available without prescription and often used for treatment of fever and pain. An extensive research of the literature published in analytical and pharmaceutical chemistry journals has been conducted and the chromatographic methods which were used for the purity, stability and pharmacokinetic studies of the cyclooxygenase-2 inhibitors, in formulations and biological materials have been reviewed. The methodology for the analysis of selected drugs is very well documented and many examples are available in the literature. The common use of chromatographic techniques with various detection attachments provide possibility for monitoring of drugs in therapy.

Several methods to determine amino acids and vitamins in biological and pharmaceutical samples have been reported. Thin layer chromatography (TLC) finds its place when the relatively costly equipment required by other methods is unavailable. This review covers the 1991–2010 literature on TLC/HPTLC (high performance thin layer chromatography) amino acid and vitamin determinations. It gives an overview of the special features as well as the problems in TLC/HPTLC determinations of amino acids and vitamins. Various chromatographic systems useful in amino acid and vitamin identification, separation and quantitation of are presented in tabular form. Future prospects of TLC/HPTLC for amino acid and vitamin determinations are also discussed.

One of the most promising separation techniques that have emerged during the last decade is based on the use of molecularly imprinted polymers (MIPs). MIPs are stable polymers that possess specific cavities designed for a template molecule, endowed with excellent selectivity compared to regular solid phase extraction techniques. Molecularly imprinted solid-phase extraction (MISPE) has already shown a high efficiency for the sample preparation from complex matrices. Natural products received huge attention in recent years. Indeed, the application of MISPE for the screening of natural products appears extremely interesting not only for the selective extraction of a target compound but also for the concomitant discovery of new drug candidates, promising sources of therapeutic benefits. In the present review, examples of recognition and separation of active components from natural extracts are emphasized. MIPs are very promising materials to mimic the recognition characteristics exhibited by enzymes or receptors although further developments are necessary to fully exploit their wide potential.

Several natural and synthetic substances have been monitored in analytical laboratories worldwide to ensure food safety. Multiple residue detection (i.e., detection of multiple analytes in a single sample or matrix) is a main weakness of existing analytical methods, when fast and reliable results are required. Multianalyte approaches may save time and money in the food industry, and more importantly, they allow the quick release of food products into the marketplace. In addition, multianalyte approaches notably decrease the time required between sampling and analysis to meet legal requirements. However, to achieve analytical success, it is necessary to develop thorough clean-up procedures to extract analytes from the matrix. In addition, good chromatographic separation methods are also necessary to distinguish closely related analytes. Molecular imprinting technology (MIT) is an emerging, powerful tool for sample extraction and chromatography. First used for solid-phase extraction, molecularly imprinted polymers (MIPs) are also effective chromatographic phases for the separation of isomers and structurally related molecules. In recent years, a number of analytical methods utilising MIT have been applied for the analysis of residues in food, and existing methodologies have been improved. This review article describes the latest applications of MIT in the development of methodologies to monitor the presence of residues of veterinary products in foodstuff.

Carbon nanotubes are among the plethora of novel nanostructures developed since the 1980s. Nanotubes have attracted considerable interest by the scientific community thanks to their extraordinary physical and chemical properties. Research areas have flourished in recent years and now include the nano-electronic, (bio)sensor and analytical field along with many others. This review covers applications of carbon nanotubes in capillary electrophoresis, capillary electrochromatography and microchip electrophoresis. First, carbon nanotubes and a range of electrophoretic techniques are briefly introduced and key references are mentioned. Next, a comprehensive survey of achievements in the field is presented and critically assessed. The merits and downsides of carbon nanotube addition to the various capillary electrophoretic modes are addressed. The different schemes for fabricating electrochromatographic stationary phases based on carbon nanotubes are discussed. Finally, some future perspectives are offered.

Alkaloids are biologically active compounds widely used as pharmaceuticals and synthesised as secondary methabolites in plants. Many of these compounds are strongly toxic. Therefore, they are often subject of scientific interests and analysis. Since alkaloids — basic compounds appear in aqueous solutions as ionized and unionized forms, they are difficult for chromatographic separation for peak tailing, poor systems efficiency, poor separation and poor column-to-column reproducibility. For this reason it is necessity searching of more suitable chromatographic systems for analysis of the compounds. In this article we present an overview on the separation of selected alkaloids from different chemical groups by liquid chromatography thus indicating the range of useful methods now available for alkaloid analysis. Different selectivity, system efficiency and peaks shape may be achieved in different LC methods separations by use of alternative stationary phases: silica, alumina, chemically bonded stationary phases, cation exchange phases, or by varying nonaqueous or aqueous mobile phase (containing different modifier, different buffers at different pH, ion-pairing or silanol blocker reagents). Developments in TLC (NP and RP systems), HPLC (NP, RP, HILIC, ion-exchange) are presented and the advantages of each method for alkaloids analysis are discussed.

Proteomic analysis has been widely used in elucidating the mechanism of diseases. As a classical proteomic approach, two-dimensional gel electrophoresis (2DGE) has been commonly applied in finding differentially expressed proteins through a first dimension of separation by the isoelectric point (pI) of proteins and a second dimension of separation according to the molecular weight (MW) of proteins. Compared to 2DGE, a recently developed commercial system from Beckman Coulter, the two-dimensional protein fractionation (PF2D), separates proteins according to the pI of proteins in the first dimension followed by a second dimension of separation according to the degree of protein hydrophobicity. As a liquid-based fractionation system, PF2D could facilitate the extraction and separation of broader protein categories and improve reproducibility and quantification as well as be less labor-intensive, which are usually identified as limitations of a gel-based 2DGE platform. This review evaluates the applications of the PF2D system and discusses the perspectives and advantages of PF2D in the investigation of cancer and genetic disorders and in protein mapping in human biological fluids and cell cultures.

A theory of comprehensive two-dimensional separations by liquid chromatographic techniques is overviewed. It includes heart-cutting and comprehensive two-dimensional separation modes, with attention to basic concepts of two-dimensional separations: resolution, peak capacity, efficiency, orthogonality and selectivity. Particular attention is paid to the effects of sample structure on the retention and advantages of a multi-dimensional HPLC for separation of complex samples according to structural correlations. Optimization of 2D separation systems, including correct selection of columns, flow-rate, fraction volumes and mobile phase, is discussed. Benefits of simultaneous programmed elution in both dimensions of LCxLC comprehensive separations are shown. Experimental setup, modulation of the fraction collection and transfer from the first to the second dimension, compatibility of mobile phases in comprehensive LCxLC, 2D asymmetry and shifts in retention under changing second-dimension elution conditions, are addressed. Illustrative practical examples of comprehensive LCxLC separations are shown.

Residues of pesticides, pharmaceutical and personal care products (PPCPs) are contaminants of world-wide concern. Consequently, there is a growing need to develop reliable analytical methods, which enable rapid, sensitive and selective determination of these pollutants in environmental samples, at trace levels. In this paper, a review of the liquid chromatography-tandem mass spectrometry (LC-MS/MS) based methods for the determination of pesticides and PPCPs in the environment is presented. Advanced aspects of current LC-MS/MS methodology, including sample preparation and matrix effects, are discussed.

The pesticide residues in foods have received increasing attention as one of the most important food safety issues. Therefore, more strict regulations on the maximum residue limits (MRLs) for pesticides in foods have been established in many countries and health organizations, based on the sensitive and reliable analysis methods of pesticide residues. However, the analysis of pesticide residues is a continuing challenge mainly because of the small quantities of analytes as well as the large amounts of interfering substances which can be co-extracted with them, often leading to experimental errors and damage to the analytical instruments. Thus, extensive sample preparation is often required for the pesticide residue analysis for the effective extraction of the analytes and removal of the interferences. This paper focuses on reviewing the recent development in the sample preparation methods for the pesticide residue analysis in foods since 2006. The methods include: liquid-liquid extraction (LLE), supercritical-fluid extraction (SFE), pressurized-liquid extraction (PLE), microwave-assisted extraction (MAE), ultrasound-assisted extraction (UAE), gel permeation chromatography (GPC), solid-phase extraction (SPE), molecularly imprinted polymers (MIPs), matrix solid-phase dispersion (MSPD), solid-phase micro-extraction (SPME), QuEChERS, cloud point extraction (CPE) and liquid phase micro-extraction (LPME), etc. Particularly their advantages, disadvantages and future perspectives will be discussed.