This issue covers a plurality of topics ranging from gold nanoparticles in separations, over size exclusion chromatography, micellar capillary electrophoresis, and affinity chromatography to monolithic columns, attracting large numbers of readers as inferred from numerous hits at the JSS website and many downloads.
Recent advances in application of hydrophilic interaction chromatography for quantitative bioanalysis
Wenying Jian, Richard W. Edom, Yaodong Xu, and Naidong Weng
HILIC has been proven to be a powerful tool for separation of polar compounds and has afforded increased selectivity, higher sensitivity, and improved efficiency for quantitation of drug and their metabolites in complex biological matrices. Practical technique considerations, orthogonal separations, fast separations, and overcoming matrix effects using novel techniques, will be highlighted and their impact on quantitative bioanalysis will be discussed.
Keywords: Bioanalysis, Body fluids, Drug metabolism, HILIC-MS, On-line extraction, Polar compounds, Quantitation
On-line desorption of dried blood spots coupled to hydrophilic interaction/reversed-phase LC/MS/MS system for the simultaneous analysis of drugs and their polar metabolites
Aurélien Thomas, Julien Déglon, Thierry Steimer, Patrice Mangin, Youssef Daali, and Christian Staub
Simultaneous analysis of polar and non-polar compounds remains a challenge to many applications. The use of the dried blood spot technique is particularly interesting since it significantly reduces the blood volume sampling. The aim of this work is to present a complete procedure coupling the on-line DBS desorption process with an automated system for monitoring compounds with a wide range of polarity.
Keywords: Body fluids, Drug metabolism, HILIC-RP, Non-polar compounds, On-line DBS process, Polar compounds, Whole blood
Visible light initiated polymerization of styrenic monolithic stationary phases using 470 nm light emitting diode arrays
Zarah Walsh, Pavel A. Levkin, Vijay Jain, Brett Paull, Frantisek Svec, and Mirek Macka
Poly(styrene-co-divinylbenzene) monolithic stationary phases have been synthesized using photoinduced polymerization. Initiation with visible light at 470 nm could be conveniently achieved with an LED array. This new development will allow more flexibility within the area of photo-induced polymerization of monolithic stationary phases.
Keywords: LED, Monolith, Photoinitiated polymerization, Stationary phases, Styrene, Visible light
A new thermally immobilized fluorinated stationary phase for RP-HPLC
Liane Maldaner and Isabel C. S. F. Jardim
The continuous development of new types of stationary phases focuses principally on novel selectivities required for the separation of complex mixtures and on improving the peak shapes of basic compounds. A new fluorinated SP, based on poly-(methyl-3,3,3-trifluoropropylsiloxane) (PMTFS), was prepared by thermal immobilization of this polymer onto silica. The fluorinated SP present different characteristics and can be employed for the separation of basic compounds and fluorinated positional isomers.
Keywords: Experimental design, HILIC-RP, Poly(methyl-3,3,3-trifluoropropylsiloxane), Stationary phases, Thermal immobilization
Knowledge discovery in metabolomics: An overview of MS data handling
Julien Boccard, Jean-Luc Veuthey, and Serge Rudaz
As continued data accumulation is inevitable, finding automatically the valuable but hidden information in metabolomic data of high dimensionality is already a crucial issue. The merging with data from other Omic technologies will probably offer a deeper insight into the regulatory networks in systems biology and will definitely represent another great challenge. Modeling all types of regulatory events occurring within a cell in a holistic manner is expected to provide a global view of the system.
Keywords: Data mining, Data processing, Hierarchical clustering, Metabolomics, Systems biology
Hydrophilic interaction chromatography coupled to MS for metabonomic/metabolomic studies
Konstantina Spagou, Helen Tsoukali, Nikolaos Raikos, Helen Gika, Ian D. Wilson, and Georgios Theodoridis
This review describes the applications and the utility of HILICMS in metabolomic/metabonomic studies and highlights certain characteristic examples in the life and plant-food sciences. HILIC is a very attractive approach in the field of metabolite analysis where it is essential that the analytical technique extracts as much information as possible from the analysis of samples of varying nature (from polar biological fluids to organic extracts of plant or biological tissue).
Keywords: Biomarkers, Metabolomics, Metabonomics, Polar compounds
Evaluation of various HILIC materials for the fast separation of polar compounds
Bénédicte Chauve, Davy Guillarme, Philippe Cléon, and Jean-Luc Veuthey
Regarding the kinetic performance of the fast-HILIC columns, two technologies were compared: the use of columns packed with sub-2 mm porous particles (UHPLC) and columns packed with sub-3 mm superficially porous particles (fused core). The choice of the dissolution solvent was critical to obtain suitable peak shape in fast HILIC separations with a composition as close as possible to the mobile-phase composition.
Keywords: Drug metabolism, Kinetic plots, Sub-2 µm particles, Superficially porous particles, UPLC
Multidimensional chromatography coupled to mass spectrometry in analysing complex proteomics samples
Péter Horvatovich, Berend Hoekman, Natalia Govorukhina, and Rainer Bischoff This review describes important aspects and characteristics of LCn-MS analysis platforms focusing on complex proteomics samples. It provides an overview of the most recent techniques, methodological developments, platform performance, chemometrics and bioinformatics methods to process LCn-MS data for comprehensive differential protein expression profiling.
Keywords: Bioinformatics, Data processing, LCn-MS, Multidimensional separation, Proteomics; Quantitation
Hydrophilic interaction LC of peptides: Columns comparison and clustering
Sylvia Van Dorpe, Valentijn Vergote, Adel Pezeshki, Christian Burvenich, Kathelijne Peremans, and Bart De Spiegeleer
Different chromatographic response criteria as well as different statistical chemometrical techniques were used for HILIC column clustering and classification based on the chromatographic analysis of ten pharmaceutically relevant model peptides.
Keywords: Classification, Hierarchical clustering, Peptides, Principal component analysis, Stationary phases
Protein glycosylation analysis by HILIC-LC-MS of Proteinase K-generated N- and O-glycopeptides
Gerhild Zauner, Carolien A. M. Koeleman, André M. Deelder, and Manfred Wuhrer
Here, a method for the MS characterization of site-specific protein glycosylation is presented. Using asialofetuin and fetuin as model substances, a protocol for glycopeptide dissection was developed based on unspecific proteolysis by Proteinase K.
Keywords: Glycopeptides, Glycosylation, Nano ESI-MS, Proteinase K
Characterization of stationary phases for reversed-phase chromatography
Martina Molíková and Pavel Jandera
Classical chromatographic column performance tests are compared with tests based on the selectivity for non-polar alkylbenzenes and with tests based on the isomer selectivity for naphthalene sulphonic acids. It is difficult to characterize the suitability of the individual columns for general HPLC applications, as the suitability of each commercial column depends on the type of sample and on the aim of separation.
Keywords: Column characterization, Homologues series, Lipophilicity, Naphthalene sulphonic acids, Silanol activity
Enhanced glyco-profiling by specific glycopeptide enrichment and complementary monolithic nano-LC (ZIC-HILIC/RP18e)/ESI-MS analysis
Jessica Wohlgemuth, Michael Karas, Wen Jiang, Robertus Hendriks, and Sven Andrecht
Monitoring of protein glycosylation has become a very important issue and requires specific analytical tools and methodologies for elucidation of the entire structural heterogeneity in relation to its biological function. It could be demonstrated that a combination of glycopeptides batch enrichment with HILIC and RP mode LC prior to MS analysis facilitates significantly enhanced glycopeptides profiling and detection of a higher number of different and low-abundant glycoforms.
Keywords: Batch enrichment, Glycopeptides, Glycosylation, Monoliths
Size exclusion chromatography – a blessing and a curse of science and technology of synthetic polymers
The size exclusion chromatography method is presented from both its positive and its adverse sides. It is shown that this outstanding, extremely useful, simple, robust, and repeatable method may produce erroneous results if appropriate attention is not paid to the conditions of measurement and to the data processing.
Keywords: Data processing, Macromolecules, Practical separation science, Synthetic polymers
Capillary electrophoresis applied to proteomic analysis
Bryan R. Fonslow and John R. Yates III
In the postgenomic era, proteomics has become a dominant field for idenying and quantifying the complex protein machinery of the cell. Here, the impact of capillary electrophoresis as a powerful separation method in proteomics will be reviewed, particularly in proven methods, relevant developments on model protein and peptide systems, and biological and clinical application, followed by comments on potential advances and applications.
Keywords: Enrichment, Microscale, Proteomics
Application of gold nanoparticles in separation sciences
David Sýkora, Václav Kašička, Ivan Mikšík, Pavel Řezanka, Kamil Záruba, Pavel Matějka, and Vladimír Král
This review article solely focuses on gold nanoparticles with the emphasis imposed upon their application in the separation arena, especially in electromigration and chromatographic techniques. In addition, typical preparation procedures of the GNPs and their modifications are presented and physico-chemical and analytical methods employed for characterization of the native and modified GNPs are briefly described.
Keywords: DNA fragments, Gold nanoparticles, Microscale, Proteins
Sample preparation and fractionation for proteome analysis and cancer biomarker discovery by mass spectrometry
Farid E. Ahmed
The field of sample preparation for proteomics is still in its infancy. There is no general standardized strategy for overall sample preparation, separation or purification. Problems associated with general and specialized strategies of sample preparation and fractionation, dealing with samples that are solution or suspension, in a frozen tissue state, or formalin-preserved tissue archival samples, and illustrates how sample processing might influence detection with mass spectrometric techniques.
Keywords: Bioanalysis, Body fluids, Morphine, Drug metabolism, Opium, Polar compounds
Less common applications of monoliths: IV. Recent developments in immobilized enzyme reactors for proteomics and biotechnology
Jana Krenkova and Frantisek Svec
This review summarizes the literature published since 2006 and demonstrates the broad variability of reactive monoliths prepared from silica as well as from organic polymers in the form of disks, columns, and capillaries. Although some of the new developments are just refinements of the methods developed previously, some notable new approaches have also been reported.
Keywords: Biotechnology, Enzyme reactors, Immobilization, Monoliths, Proteomics
Bioaffinity chromatography on monolithic supports
Kishore K. R. Tetala, Teris A. van Beek
Selective enrichment studies using either bioaffinity or immunoaffinity chromatography on monolithic supports indicates that this field is still very much alive if not expanding. This review starts off with an introduction on affinity chromatography, followed by the types of ligands, types of monolithic supports and their preparation. Individual real-life applications are discussed in detail.
Keywords: Bioaffinity, Enrichment, Immunoaffinity, Immobilization, Ligands, Monoliths