Download or read book The Analysis of Biological Samples Using Capillary Electrophoresis and HPLC Technologies written by and published by . This book was released on 2007 with total page 201 pages. Available in PDF, EPUB and Kindle. Book excerpt: Capillary electrophoresis (CE) and high performance liquid chromatography (HPLC) are two very important separation techniques. CE is a microseparation technique in which an electric field is applied to perform the separation. In CE, charged species are separated based on their charge/mass ratio, making this technique useful for separating a wide variety of components, including those found in biological samples. HPLC is the most widely used separation technique due to the ease of use and wide variety of components that can be separated. HPLC employs the use of pressure to push mobile phase through an analytical column containing a stationary phase that interacts with the solutes to effect the separation. Both CE and HPLC can use a number of different detection schemes to monitor the separated species, including ultra-violet (UV), laser-induced fluorescence (LIF), and mass spectrometry (MS). Affinity capillary electrophoresis (ACE) combines the separation power of CE with the selectivity of immunoassays. A fluorescein labeled estradiol derivative was evaluated for potential use as a competitive inhibitor in ACE. The labeling procedure rendered multiple components with the fluorescein label. Two of the components were identified as active estradiol species against the anti-estradiol antibody, examined by CE-LIF. Since only one active species had been predicted, both species were characterized using CE-MS and MS/MS, in addition to testing them in the ACE format. Nucleoside reverse transcriptase inhibitors (NRTIs) are one class of anti-HIV (human immunodeficiency virus) drugs. Because HIV is considered a pandemic, the analysis of NRTIs in human samples is very important; people all over the world are using these drugs. Currently, multiple NRTIs are given for treatment; however, there are few methods to date that separate and detect multiple NRTIs in a single run. NRTIs are metabolized in the body into their active triphosphate form. Two approaches can be followed for their separation and detection. The first approach was an indirect method, in which the active triphosphate form was dephosphorylated into the parent form of the NRTIs. HPLC with UV detection was utilized to establish an optimized separation method prior to using LC-MS/MS for the analysis of the following NRTIs in human peripheral blood monunclear cells (PBMCs): stavudine (D4T), lamivudine (3TC), the metabolized form of didanosine (ddA), zidovudine (ZDV), and the internal standard. The second approach was a direct method, in which analysis of the triphosphate form of the NRTIs was performed without dephosphorylation of the drugs. A capillary eletrophoretic method was developed that allows separation of the triphosphate NRTIs of interest: zidovudine triphosphate (ZDV-TP), stavudine triphosphate (D4T-TP), lamivudine triphosphate (3TC-TP), 2',3'-dideoxyadenoine triphosphate (ddA-TP), tenofivir diphosphate (TFV-DP), and emtricitabine triphosphate (FTC-TP). The separation was optimized using UV detection. Since UV detection does not offer appropriate detection limits, we studied the native fluorescence spectroscopic characteristics of the triphosphate NRTIs. We constructed an in-house CE-LIF system to explore LIF detection of the phosphorylated NRTIs. The system employed the use of a deep-UV laser with excitation at 224 nm. A sheath-flow cuvette was integrated into the design to improve detectability. The CE-LIF system was aligned and tested using the native fluorescence of tryptophan. The CE-LIF system was then used for the analysis of the active metabolized forms of NRTIs. With this system, we were able to separate and detect ZDV-TP, D4T-TP, 3TC-TP, ddA-TP, and the internal standard using a modification of the separation conditions previously optimized with UV detection. The detection limits obtained are not sufficiently low for the analysis of human PBMCs.
Download or read book The Analysis of Biological Samples by Capillary Electrophoresis written by and published by . This book was released on 2007 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Capillary Electrophoresis Mass Spectrometry written by James Q. Xia and published by Springer. This book was released on 2016-11-08 with total page 80 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book covers the latest developments in capillary electrophoresis-mass spectrometry for the analysis of therapeutic proteins. The application of capillary electrophoresis-mass spectrometry (CE-MS) coupling technology in the analysis of recombinant therapeutic proteins is detailed thoroughly. Specific topics include recent developments in coupling capillary electrophoresis with mass spectrometry for the quality control of monoclonal antibody therapeutics, top-down analysis of monoclonal antibody using the CE-MS platform, and detection of host cell protein impurities. Comprehensive characterization of antibody-drug conjugates (ADCs) by coupling capillary electrophoresis with mass spectrometry is also covered. This is an ideal book for scientists in the life science and biopharmaceutical industry who are working on characterizing the PTMs of monoclonal antibodies, as well as graduate students and researchers in the separation science and biological mass spectrometry fields.
Download or read book High Performance Liquid Chromatography Capillary Electrophoresis written by Andrea Weston and published by Elsevier. This book was released on 1997-07-21 with total page 295 pages. Available in PDF, EPUB and Kindle. Book excerpt: HPLC and CE: Principles and Practice presents the latest information on the most powerful separation techniques available: high-performance liquid chromatography (HPLC) and capillary electrophoresis (CE). Fundamental theory, instrumentation, modes of operation, and optimization of separations are presented in a concise, non-technical style to help the user in choosing the appropriate technique quickly and accurately. Well- illustrated and containing convenientend-of-chapter summaries of the major concepts, the book provides in-depth coverage of trouble-shooting, improvement of resolution, data manipulation, selectivity, and sensitivity.Graduate students, technicians, and researchers who must use separations with little or no background in analytical chemistry can overcome separation anxiety and get started in obtaining the best possible separations in minimal time. The book will alsobe useful to analytical chemists who need a better understanding of theory and processes. Fully up-to-date information on both HPLC and CE includes troubleshooting and comparisons of the two techniques Applicable to a wide variety of separation problems Covers basic concepts governing any separation as well as instrumentation and how to use it Helps the user to obtain optimal resolution in minimal time Contains information on special procedures such as chiral separations, affinity chromatography, and sample preparation Includes information on upcoming trends such as miniaturization Major concepts in each chapter are organized to allow access to information easily and quickly Contains practical bibliography for accessing the literature
Download or read book Capillary Electrophoresis Mass Spectrometry for Metabolomics written by Rawi Ramautar and published by Royal Society of Chemistry. This book was released on 2018-07-13 with total page 300 pages. Available in PDF, EPUB and Kindle. Book excerpt: Capillary electrophoresis–mass spectrometry (CE-MS) has become a very useful analytical technique for the profiling of highly polar and charged metabolites in biological samples. In this book, the unique features of CE-MS for metabolomics studies are highlighted including CE separation modes, capillary coatings and practical aspects of CE-MS coupling alongside a comprehensive overview of recent technological developments and applications. CE-MS can be considered a relatively new technique in the field of metabolomics and it is therefore important to inform the scientific community about the possibilities of advanced CE-MS approaches for metabolomics studies. This book outlines the potential of this technique for researchers working in metabolomics, bioanalytics and biomarker analysis.
Download or read book Development of Analytical Protocols for Rapid Analysis of Biological Samples with Flow gated Capillary Electrophoresis written by Ning Zhang and published by . This book was released on 2023 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: With the development and improvement of the whole society, people are paying more attention to their health and the protection of the environment, which requires researchers to consider better identification and detection methods for various molecular species. Capillary electrophoresis (CE) and related technologies have been used in the separation, detection, and identification of many organic and inorganic compounds for the past dozens of years because of the advantages of CE, like short analysis time and high separation efficiency. Also, numerous detectors and interfaces can be combined with CE due to the excellent adaptability in order to enhance the performance of detection sensitivity, e.g., laser-induced fluorescence (LIF), mass spectrometry (MS), and electrochemical detection. This thesis is to discuss the determination of essential biomarkers in biological samples on the flow-gated CE-LIF system. It first reports a rapid and sensitive method with the flow-gated CE-LIF system to determine collagen degradation products, including proline, hydroxyproline, and other prolyl dipeptides in human urine samples, which is suited for the early diagnosis of collagen-related diseases. Experimental results prove that this method was capable of the separation and determination of prolyl compounds with good reproducibility, accuracy, sensitivity, and robustness. Second, the utilization of solid-phase extraction (SPE) in the purification and concentration of catecholamines (CAs) highlights the effectiveness of this method as a sample pre-treatment technique for catecholamines in real samples. By effectively removing interfering compounds and enhancing the concentration of the target analytes, SPE contributes to the overall reliability and accuracy of CA analysis.
Download or read book Analytical Biotechnology written by Csaba Horváth and published by . This book was released on 1990 with total page 232 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume presents accounts of some of the recent advances in high performance liquid chromatography and capillary electrophoresis with regard to biotechnology. Four of its 11 chapters present an introduction to capillary electrophoresis and discuss its application to various analytical problems ranging from the analysis of cyclic nucleotides to quality control in the pharmaceutical industry. Subsequent chapters cover recent developments in HPLC, with emphasis on analysis of pharmaceutical proteins; the problems associated with the use of HPLC as a detection method in preparative chromatography; the use of mass spectrometry in the structure determination of peptides; and the use of the displacement mode of chromatography.
Download or read book Capillary Electrophoresis and Microchip Capillary Electrophoresis written by Carlos D. García and published by John Wiley & Sons. This book was released on 2013-02-01 with total page 638 pages. Available in PDF, EPUB and Kindle. Book excerpt: Explores the benefits and limitations of the latest capillary electrophoresis techniques Capillary electrophoresis and microchip capillary electrophoresis are powerful analytical tools that are particularly suited for separating and analyzing biomolecules. In comparison with traditional analytical techniques, capillary electrophoresis and microchip capillary electrophoresis offer the benefits of speed, small sample and solvent consumption, low cost, and the possibility of miniaturization. With contributions from a team of leading analytical scientists, Capillary Electrophoresis and Microchip Capillary Electrophoresis explains how researchers can take full advantage of all the latest techniques, emphasizing applications in which capillary electrophoresis has proven superiority over other analytical approaches. The authors not only explore the benefits of each technique, but also the limitations, enabling readers to choose the most appropriate technique to analyze a particular sample. The book's twenty-one chapters explore fundamental aspects of electrophoretically driven separations, instrumentation, sampling techniques, separation modes, detection systems, optimization strategies for method development, and applications. Specific topics include: Critical evaluation of the use of surfactants in capillary electrophoresis Sampling and quantitative analysis in capillary electrophoresis Capillary electrophoresis with electrochemical detection Overcoming challenges in using microchip electrophoresis for extended monitoring applications Capillary electrophoresis of intact unfractionated heparin and related impurities Microchip capillary electrophoresis for in situ planetary exploration Each chapter begins with an introduction and ends with conclusions as well as references to the primary literature. Novices to the field will find this book an easy-to-follow introduction to core capillary electrophoresis techniques and methods. More experienced investigators can turn to the book for troubleshooting tips and expert advice to guide them through the most advanced applications.
Download or read book HPLC in Enzymatic Analysis written by Edward F. Rossomando and published by John Wiley & Sons. This book was released on 2009-09-25 with total page 480 pages. Available in PDF, EPUB and Kindle. Book excerpt: The use of High Performance Liquid Chromatography (HPLC) techniques in the study of enzymatic reactions has grown significantly since the publication of the first edition of this highly successful book: the role of enzymes in biological research has expanded; the application of HPLC and enzymes has extended to more disciplines; advances in separation techniques and instrumentation have increased the capability of HPLC; and the discovery of new enzymes has spawned new methods of analysis. High Performance Liquid Chromatography in Enzymatic Analysis, Second Edition addresses these developments in its coverage of the refinements of HPLC methods and their use in a wide range of laboratory applications. It offers the same practical approach found in the first edition, incorporates a wealth of new information into existing chapters, and adds new chapters to deal with new applications, including capillary electrophoresis, forensic chemistry, microdialysis, and the polymerase chain reaction. Topics include: * Application of HPLC to the assay of enzymatic activities * Concepts and principles of HPLC, including the latest technological advances * Concepts and principles of capillary electrophoresis (CE) * Strategy for design of an HPLC/CE system for assay of enzyme activity * Preparation of enzymatic activities from tissues and single cells * Analysis of enzymatic activities in body fluids, including chromatobiosis * HPLC for the identification of new enzymatic activities * Fundamentals of the polymerase chain reaction * HPLC in forensics * Survey of enzymatic activities assayed by the HPLC method, including many new categories * Multienzyme systems, including many new examples * HPLC in the analysis of contaminated food "It is the ability of HPLC to accomplish separations completely and rapidly that led to its original application to problems in the life sciences, particularly those related to purification. An analysis of the literature revealed that this technique was used primarily for the purification of small molecules, macromolecules such as peptides and proteins, and more recently, antibodies. This application to purification has all but dominated the use of the method, and there has been a plethora of books, symposia, and conferences on the use of HPLC for these purposes. However, it was only a matter of time before others began to look beyond and to explore the possibilities that result from the capacity to make separations quickly and efficiently." --from the preface to the First Edition Easy to read and full of practical advice and hundreds of diagrams and examples, High Performance Liquid Chromatography in Enzymatic Analysis, Second Edition is an invaluable resource for students, researchers, and laboratory workers in analytical chemistry and biochemistry, molecular biology and cell biology, and for anyone interested in keeping up with this fast-growing field.
Download or read book Analysis of Small Molecules in Biological Samples Using Capillary Electrophoresis written by Zhaohui Chen and published by . This book was released on 2001 with total page 454 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Chiral Capillary Electrophoresis in Current Pharmaceutical and Biomedical Analysis written by Peter Mikuš and published by BoD – Books on Demand. This book was released on 2012-08-29 with total page 211 pages. Available in PDF, EPUB and Kindle. Book excerpt: The scientific monograph by the author Peter Mikus entitled "Chiral Capillary Electrophoresis in Current Pharmaceutical and Biomedical Analysis" provides a comprehensive view on the advanced capillary electrophoresis techniques aimed to current chiral bioanalysis. The advances in the chiral electrophoresis analytical approaches are divided and theoretically described in three sections involving (i) advanced chiral separations for the optimization of chiral resolution (separation mechanisms; electrophoresis techniques in capillary and microchip format; electrophoretic modes such as ITP, CZE/EKC, CEC; chiral additives / pseudophases / phases), (ii) advanced sample preparation for the on-line preconcentration, sample clean-up and analyte derivatization (implementation of electrophoretic effects such as stacking; non-electrophoretic effects such as SPE, chromatography, dialysis; combinations of these effects; multidimensional CE systems; instrumental schemes), (iii) advanced combinations of detection and electrophoresis for the optimization in qualitative and quantitative evaluation (the most important universal as well as selective detection approaches such as absorption and fluorescence spectrophotometry, electrochemical detection, mass spectrometry vs. (i) and/or (ii)). Real analytical potential (benefits and limitations) of these advanced analytical approaches is emphasized by selected performance parameters of the methods and illustrated by many current practical applications including chiral analyses of drugs, their (bio)degradation products and biomarkers in pharmaceutical and biological matrices. The author wishes the readers many inspirations in the creation of new innovative approaches in the field of advanced chiral electrophoresis techniques with the aim to overcome capabilities of the current analytical techniques.
Download or read book Chromatography and Capillary Electrophoresis in Food Analysis written by Hilmer Sorensen and published by Royal Society of Chemistry. This book was released on 2007-10-31 with total page 489 pages. Available in PDF, EPUB and Kindle. Book excerpt: In the rapidly developing field of analysis it is important to be aware of the newest methods within available techniques. Chromatography and Capillary Electrophoresis in Food Analysis describes chromatographic and electrophoretic principles and procedures for analyses of various amphiphilic and hydrophilic biomolecules, particularly for food analysis. Providing basic information, including general sample preparation, the book then goes on to describe individual analytical methods and exemplify the strategy and methodologies employed for the analyses. The theory necessary to understand the methods and interpretation of results is also included, as are numerous detailed instructions on experiments. Tables, figures and references are included to give a complete picture. Chromatography and Capillary Electrophoresis in Food Analysis will be especially valuable for students and more experienced researchers interested in analysis of natural products, both inside and outside the field of food chemistry.
Download or read book Liquid Chromatography and Capillary Electrophoresis Methodologies for the Analysis of Biological Samples written by and published by . This book was released on 2008 with total page 256 pages. Available in PDF, EPUB and Kindle. Book excerpt: Separation techniques such as capillary electrophoresis and liquid chromatography have found application in the analysis of biological samples. In capillary electrophoresis charged species are separated based on their mass to charge ratios under the influence of an applied electric field. In liquid chromatography, analytes are separated based on their partition between a stationary and mobile phase. Separation techniques when combined with a variety of detection techniques such as ultraviolet, laser induced fluorescence, mass spectrometry and electrochemical detection have been very successful in the quantification of analytes comprised in complex mixtures (e.g., biological samples). Capillary electrophoresis (CE) with laser induced fluorescence (LIF) detection was used to separate different bioconjugated CdSe/ZnS quantum dots (QDs). The QD nanocrystals studied were conjugated to the biomolecules streptavidin, biotin, and immunoglobulin G. The bioconjugated QDs showed different electrophoretic mobilities, which appear to depend upon the biomolecule that is attached to the QD and the buffer solution used. The use of a polymeric additive into the CE run buffer improved the resolution of the bioconjugates. Under CE conditions, the interaction between QD bioconjugates containing streptavidin (QDSt) and biotin (QDBi) was monitored. In a two color experiment, two different sizes of QD (i.e., 585 nm and 655 nm) were used to monitor the interaction between 655 nm QDBi and 585 nm QDSt. The use of QDs with different emission properties allows the selective monitoring of two different wavelengths while using one single excitation source. This in turn, allowed the monitoring of overlapping peaks in the electropherogram when newly formed products resulting from the interaction of the two bioconjugated QDs appeared. Protease inhibitors (PIs) are a class of compounds that are used in the treatment of patients with the human immunodeficiency virus (HIV). These drugs inhibit the HIV-1 protease, acting on the production of viral proteins essential for the completion of the viral replication process. Monitoring free drug concentration is necessary to establish the dosage that would result in optimum free drug concentration in blood for a given patient. We developed methodology to analyze free protease inhibitor lopinavir in plasma samples. Ultrafiltration was used to isolate the free drug; then LC/MS/MS was used for sample analysis. Restrictive access media columns have been developed to simplify sample preparation processes. These columns allow the direct injection of untreated biological samples. Restrictive access media columns exclude macromolecules such as proteins, while retaining small molecules such as drugs. Two restrictive access material columns were evaluated for the direct injection of plasma for analysis of the protease inhibitor lopinavir. In the columns studied the interaction of plasma components with the material inside the column was more than expected. Glucose monitoring and insulin therapy are becoming a standard of care in patients in the intensive care unit. In order to control glycemic levels, frequent glucose monitoring is required. The use of tear fluid glucose concentrations was evaluated as a non-invasive sampling strategy for monitoring glucose in intensive care unit patients. Glucose in tear fluid samples from intensive care unit patients were analyzed using anion exchange chromatography with pulsed amperometric detection. The results did not show a correlation between tear fluid and blood glucose concentrations.
Download or read book Capillary Electrophoresis Mass Spectrometry for Proteomics and Metabolomics written by Rawi Ramautar and published by John Wiley & Sons. This book was released on 2022-10-03 with total page 404 pages. Available in PDF, EPUB and Kindle. Book excerpt: Capillary Electrophoresis—Mass Spectrometry for Proteomics and Metabolomics A powerful and essential resource for researchers with an interest in CE-MS In Capillary Electrophoresis—Mass Spectrometry for Proteomics and Metabolomics: Principles and Applications, a team of distinguished researchers delivers a comprehensive overview of bioanalytical capillary electrophoresis coupled to mass spectrometry (CE-MS). The book explains foundational principles, technology as well the strategies and techniques used in data analysis for metabolic and proteomic studies. It also provides a global overview of recent developments and advances for improving CE-MS sensitivity and reproducibility. An essential handbook for everyone performing metabolomic and proteomic analysis, the information provided here will assist researchers in tapping into the full potential of this technique to answer biological and clinical questions. Readers will also find: A thorough introduction to the principles of capillary electrophoresis, including its fundamentals, CE separation modes, capillary coatings, and the fundamentals of mass spectrometry In-depth examinations of technological developments in capillary electrophoresis, including sample preparation, online preconcentration, detection sensitivity, and metabolic coverage Comprehensive discussions of metabolomic studies, including their biomedical and clinical applications Recent advances in proteomics, including top-down and bottom-up approaches Perfect for analytical and clinical chemists, Capillary Electrophoresis—Mass Spectrometry for Proteomics and Metabolomics: Principles and Applications will also earn a place in the libraries of biochemists, molecular biologists, and other molecular life scientists.
Download or read book Capillary Electrophoresis in Chiral Analysis written by Bezhan Chankvetadze and published by John Wiley & Sons. This book was released on 1997-12-29 with total page 586 pages. Available in PDF, EPUB and Kindle. Book excerpt: Das weltweite Interesse an der Trennung von Enantiomeren, Spiegelbildern ein und derselben chemischen Verbindung, nimmt zu, da diese Formen beispielsweise verschiedene physiologische Wirkungen hervorrufen können. Eine hochaktuelle Technik der Stofftrennung in diesem Zusammenhang ist die Kapillarelektrophorese, die hier von den theoretischen Grundlagen bis hin zu analytischen Details und Anbieterlisten ausführlich beschrieben wird.
Download or read book Carbohydrate Analysis written by Z. El Rassi and published by Elsevier. This book was released on 1994-11-11 with total page 693 pages. Available in PDF, EPUB and Kindle. Book excerpt: Carbohydrates and glycoconjugates play an important role in several life processes. The wide variety of carbohydrate species and their inherent polydispersity and heterogeneity require separation techniques of high resolving power and high selectivity such as high performance liquid chromatography (HPLC) and capillary electrophoresis (HPCE). In the last decade HPLC, and recently HPCE methods have been developed for the high resolution and reproducible quantitation of carbohydrates. Despite the importance of these two column separation technologies in the area of carbohydrates, no previous book describes specialized methods for the separation, purification and detection of carbohydrates and glycoconjugates by HPLC and HPCE. Therefore, the objective of the present book is to provide a comprehensive review of carbohydrate analysis by HPLC and HPCE by covering analytical and preparative separation techniques for all classes of carbohydrates including mono- and disaccharides; linear and cyclic oligosaccharides; branched heterooligosaccharides (e.g., glycans, plant-derived oligosaccharides); glycoconjugates (e.g., glycolipids, glycoproteins); carbohydrates in food and beverage; compositional carbohydrates of polysaccharides; carbohydrates in biomass degradation; etc. The book will be of interest to a wide audience, including analytical chemists and biochemists, carbohydrate, glycoprotein and glycolipid chemists, molecular biologists, biotechnologists, etc. It will also be a useful reference work for both the experienced analyst and the newcomer as well as for users of HPLC and HPCE, graduates and postdoctoral students.
Download or read book Capillary Electrophoresis in Analytical Biotechnology written by Pier Giorgio Righetti and published by CRC Press. This book was released on 1995-12-18 with total page 570 pages. Available in PDF, EPUB and Kindle. Book excerpt: This new book on capillary electrophoresis (CE) is unique in its focus on biotechnology. It is devoted to proteins, peptides, and techniques especially useful in the area of recombinant DNA products. Emphasis is also placed on glycoproteins. Because of the growing role of the glycosylation process in CE, a comprehensive chapter on the subject acts as a book within a book. Although this well-known researcher in biotechnology presents a number of chapters extensively discussing theories, important practical aspects in the routine use of capillary electrophoresis are also covered.
