WorldWideScience

Sample records for capillary electrophoresis instrumentation

  1. Serum proteins analysis by capillary electrophoresis

    OpenAIRE

    Uji, Yoshinori; Okabe, Hiroaki

    2001-01-01

    The purpose of this study was to evaluate the efficacy of multi-capillary electrophoresis instrument in clinical laboratory. An automated clinical capillary electrophoresis system was evaluated for performing serum proteins electrophoresis and immuno-fixation electrophoresis by subtraction. In this study the performance of capillary electrophoresis was compared with the cellulose acetate membrane electrophoresis and agarose gel immunofixation electrophoresis for serum proteins. The results of...

  2. Instrumental development of novel detection and separation methods for capillary electrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Garner, T.

    1993-07-01

    After a general introduction, this thesis is divided into 3 parts: indirect fluorescence detection of sugars separated by capillary zone electrophoresis with visible laser excitation, absorption detection in capillary electrophoresis by fluorescence energy transfer, and increased selectivity for electrochromatography by dynamic ion exchange.

  3. Microchip capillary electrophoresis instrumentation for in situ analysis in the search for extraterrestrial life.

    Science.gov (United States)

    Mora, Maria F; Stockton, Amanda M; Willis, Peter A

    2012-09-01

    The search for signs of life on extraterrestrial planetary bodies is among NASA's top priorities in Solar System exploration. The associated pursuit of organics and biomolecules as evidence of past or present life demands in situ investigations of planetary bodies for which sample return missions are neither practical nor affordable. These in situ studies require instrumentation capable of sensitive chemical analyses of complex mixtures including a broad range of organic molecules. Instrumentation must also be capable of autonomous operation aboard a robotically controlled vehicle that collects data and transmits it back to Earth. Microchip capillary electrophoresis (μCE) coupled to laser-induced fluorescence (LIF) detection provides this required sensitivity and targets a wide range of relevant organics while offering low mass, volume, and power requirements. Thus, this technology would be ideally suited for in situ studies of astrobiology targets, such as Mars, Europa, Enceladus, and Titan. In this review, we introduce the characteristics of these planetary bodies that make them compelling destinations for extraterrestrial astrobiological studies, and the principal groups of organics of interest associated with each. And although the technology we describe here was first developed specifically for proposed studies of Mars, by summarizing its evolution over the past decade, we demonstrate how μCE-LIF instrumentation has become an ideal candidate for missions of exploration to all of these nearby worlds in our Solar System. PMID:22965706

  4. Derivatization in Capillary Electrophoresis.

    Science.gov (United States)

    Marina, M Luisa; Castro-Puyana, María

    2016-01-01

    Capillary electrophoresis is a well-established separation technique in analytical research laboratories worldwide. Its interesting advantages make CE an efficient and potent alternative to other chromatographic techniques. However, it is also recognized that its main drawback is the relatively poor sensitivity when using optical detection. One way to overcome this limitation is to perform a derivatization reaction which is intended to provide the analyte more suitable analytical characteristics enabling a high sensitive detection. Based on the analytical step where the CE derivatization takes place, it can be classified as precapillary (before separation), in-capillary (during separation), or postcapillary (after separation). This chapter describes the application of four different derivatization protocols (in-capillary and precapillary modes) to carry out the achiral and chiral analysis of different compounds in food and biological samples with three different detection modes (UV, LIF, and MS). PMID:27645730

  5. Laser induced fluorescence and Raman spectroscopy in capillary electrophoresis as an possible instrument for extraterrestrial life signs detection.

    Science.gov (United States)

    Mikhail, Gorlenko; Cheptcov, Vladimir; Anton, Maydykovskiy; Eugeniy, Vasilev

    The one of a significant aims in extraterrestrial exploration is a seeking for a life traces in a open space and planetary objects. Complex composition and unknown origin of suspected signs of life required у new analytical approaches and technical solutions. The promising assai here can be Laser induced fluorescence and Raman spectroscopy methods. The combined instrument developed by our team reveal the advantage of capillary electrophoresis assays in a junction with laser induced fluorescence detection technology. We optimized excitation configuration of fluorescence in capillary electrophoresis to reduce pumping laser power up to 1 mW and decrease background scattering. The improvement of the device sensitivity at poor sample concentration we achieved by incorporating fluorescence flow-through cuvette into spectrometer. That allows to simplify setup, to minimize weight and increase reproducibility of measurements. The device has been tasted in complex organic chemical mixes and microbial strains differentiation tasks. 3d multinational spectra allow us to increase the spectra information loads in comparison with ordinary capillary electrophoresis approaches. Possible updating the device with Raman approach can even furthermore multiple the differentiation power of the instrument. The analytical module developed using this approach can be potentially effectively used in extraterrestrial researches as a payload of the future spacecraft.

  6. Triple-channel portable capillary electrophoresis instrument with individual background electrolytes for the concurrent separations of anionic and cationic species.

    Science.gov (United States)

    Mai, Thanh Duc; Le, Minh Duc; Sáiz, Jorge; Duong, Hong Anh; Koenka, Israel Joel; Pham, Hung Viet; Hauser, Peter C

    2016-03-10

    The portable capillary electrophoresis instrument is automated and features three independent channels with different background electrolytes to allow the concurrent optimized determination of three different categories of charged analytes. The fluidic system is based on a miniature manifold which is based on mechanically milled channels for injection of samples and buffers. The planar manifold pattern was designed to minimize the number of electronic valves required for each channel. The system utilizes pneumatic pressurization to transport solutions at the grounded as well as the high voltage side of the separation capillaries. The instrument has a compact design, with all components arranged in a briefcase with dimensions of 45 (w) × 35 (d) × 15 cm (h) and a weight of about 15 kg. It can operate continuously for 8 h in the battery-powered mode if only one electrophoresis channel is in use, or for about 2.5 h in the case of simultaneous employment of all three channels. The different operations, i.e. capillary flushing, rinsing of the interfaces at both capillary ends, sample injection and electrophoretic separation, are activated automatically with a control program featuring a graphical user interface. For demonstration, the system was employed successfully for the concurrent separation of different inorganic cations and anions, organic preservatives, additives and artificial sweeteners in various beverage and food matrices. PMID:26893094

  7. DNA Sequencing Using capillary Electrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Dr. Barry Karger

    2011-05-09

    The overall goal of this program was to develop capillary electrophoresis as the tool to be used to sequence for the first time the Human Genome. Our program was part of the Human Genome Project. In this work, we were highly successful and the replaceable polymer we developed, linear polyacrylamide, was used by the DOE sequencing lab in California to sequence a significant portion of the human genome using the MegaBase multiple capillary array electrophoresis instrument. In this final report, we summarize our efforts and success. We began our work by separating by capillary electrophoresis double strand oligonucleotides using cross-linked polyacrylamide gels in fused silica capillaries. This work showed the potential of the methodology. However, preparation of such cross-linked gel capillaries was difficult with poor reproducibility, and even more important, the columns were not very stable. We improved stability by using non-cross linked linear polyacrylamide. Here, the entangled linear chains could move when osmotic pressure (e.g. sample injection) was imposed on the polymer matrix. This relaxation of the polymer dissipated the stress in the column. Our next advance was to use significantly lower concentrations of the linear polyacrylamide that the polymer could be automatically blown out after each run and replaced with fresh linear polymer solution. In this way, a new column was available for each analytical run. Finally, while testing many linear polymers, we selected linear polyacrylamide as the best matrix as it was the most hydrophilic polymer available. Under our DOE program, we demonstrated initially the success of the linear polyacrylamide to separate double strand DNA. We note that the method is used even today to assay purity of double stranded DNA fragments. Our focus, of course, was on the separation of single stranded DNA for sequencing purposes. In one paper, we demonstrated the success of our approach in sequencing up to 500 bases. Other

  8. Biomedical applications of capillary electrophoresis

    Science.gov (United States)

    Kartsova, L. A.; Bessonova, E. A.

    2015-08-01

    The review deals with modern analytical approaches used in capillary electrophoresis for solving medical and biological problems: search for biomarkers of various diseases and rapid diagnosis based on characteristic profiles of biologically active compounds by capillary electrophoresis with mass spectrometric detection; monitoring of the residual drugs in biological fluids for evaluating the efficiency of drug therapy; testing of the enantiomeric purity of pharmaceutical products; the use of novel materials as components of stationary and pseudo-stationary phases in capillary electrophoresis and capillary electrochromatography to increase the selectivity of separation of components of complex matrices; and identification of various on-line preconcentration techniques to reduce the detection limits of biologically active analytes. A topical trend in capillary electrophoresis required in clinical practice, viz., the design of microfluidic systems, is discussed. The bibliography includes 173 references.

  9. Capillary electrophoresis in food authenticity.

    Science.gov (United States)

    Kvasnicka, Frantisek

    2005-06-01

    Food authenticity is a term which simply refers to whether the food purchased by the consumer matches its description. False description can occur in many forms, from the undeclared addition of water or other cheaper materials, or the wrong declaration of the amount of a particular ingredient in the product, to making false statements about the source of ingredients i.e., their geographic, plant, or animal origin. The aim of this review is to summarize applications of capillary electrophoresis in food authentication.

  10. Capillary Electrophoresis - Optical Detection Systems

    Energy Technology Data Exchange (ETDEWEB)

    Sepaniak, M. J.

    2001-08-06

    Molecular recognition systems are developed via molecular modeling and synthesis to enhance separation performance in capillary electrophoresis and optical detection methods for capillary electrophoresis. The underpinning theme of our work is the rational design and development of molecular recognition systems in chemical separations and analysis. There have been, however, some subtle and exciting shifts in our research paradigm during this period. Specifically, we have moved from mostly separations research to a good balance between separations and spectroscopic detection for separations. This shift is based on our perception that the pressing research challenges and needs in capillary electrophoresis and electrokinetic chromatography relate to the persistent detection and flow rate reproducibility limitations of these techniques (see page 1 of the accompanying Renewal Application for further discussion). In most of our work molecular recognition reagents are employed to provide selectivity and enhance performance. Also, an emerging trend is the use of these reagents with specially-prepared nano-scale materials. Although not part of our DOE BES-supported work, the modeling and synthesis of new receptors has indirectly supported the development of novel microcantilevers-based MEMS for the sensing of vapor and liquid phase analytes. This fortuitous overlap is briefly covered in this report. Several of the more significant publications that have resulted from our work are appended. To facilitate brevity we refer to these publications liberally in this progress report. Reference is also made to very recent work in the Background and Preliminary Studies Section of the Renewal Application.

  11. Non-Aqueous Capillary Electrophoresis

    Science.gov (United States)

    Szumski, Michał; Buszewski, Bogusław

    Non-aqueous capillary electrophoresis and capillary electrochromatography are special variants of these techniques. Here, organic solvents or their mixtures with or without dissolved electrolytes are used as separation buffer or mobile phase, respectively. The most important features of non-aqueous systems are: better solubility of more hydrophobic ionic substances (many natural products) than in water, much less current and Joule heating allows for using highly concentrated buffers and/or larger capillary internal diameters, polar interactions are enhanced in organic solvents which is often highly advantageous in chiral separation systems. This chapter presents most frequently used solvents, their properties, as well as shows pH* scale which is often used in non-aqueous systems.

  12. Capillary electrophoresis electrospray ionization mass spectrometry interface

    Science.gov (United States)

    Smith, Richard D.; Severs, Joanne C.

    1999-01-01

    The present invention is an interface between a capillary electrophoresis separation capillary end and an electrospray ionization mass spectrometry emitter capillary end, for transporting an anolyte sample from a capillary electrophoresis separation capillary to a electrospray ionization mass spectrometry emitter capillary. The interface of the present invention has: (a) a charge transfer fitting enclosing both of the capillary electrophoresis capillary end and the electrospray ionization mass spectrometry emitter capillary end; (b) a reservoir containing an electrolyte surrounding the charge transfer fitting; and (c) an electrode immersed into the electrolyte, the electrode closing a capillary electrophoresis circuit and providing charge transfer across the charge transfer fitting while avoiding substantial bulk fluid transfer across the charge transfer fitting. Advantages of the present invention have been demonstrated as effective in providing high sensitivity and efficient analyses.

  13. Electromigration dispersion in Capillary Electrophoresis

    CERN Document Server

    Chen, Zhen; 10.1007/s11538-011-9708-7

    2012-01-01

    In a previous paper (S. Ghosal and Z. Chen Bull. Math. Biol. 2010, vol. 72, pg. 2047) it was shown that the evolution of the solute concentration in capillary electrophoresis is described by a nonlinear wave equation that reduced to Burger's equation if the nonlinearity was weak. It was assumed that only strong electrolytes (fully dissociated) were present. In the present paper it is shown that the same governing equation also describes the situation where the electrolytic buffer consists of a single weak acid (or base). A simple approximate formula is derived for the dimensionless peak variance which is shown to agree well with published experimental data.

  14. Capillary electrophoresis theory and practice

    CERN Document Server

    Grossman, Paul D

    1992-01-01

    This book is designed to be a practical guide, used by wide audience, including those new to CE, those more experienced, routine users, those interested in technology development, and those involved with applications research. References have been emphasized to allow the reader to explore the detailed specifics and theoretical foundations.This book draws together the rapidly evolving, diverse, and multidisciplinary subject of capillary electrophoresis (CE). It is designed as a practical guide to be used by a wide audience, including those new to CE as well as more experienced users. T

  15. Novel absorption detection techniques for capillary electrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Xue, Y.

    1994-07-27

    Capillary electrophoresis (CE) has emerged as one of the most versatile separation methods. However, efficient separation is not sufficient unless coupled to adequate detection. The narrow inner diameter (I.D.) of the capillary column raises a big challenge to detection methods. For UV-vis absorption detection, the concentration sensitivity is only at the {mu}M level. Most commercial CE instruments are equipped with incoherent UV-vis lamps. Low-brightness, instability and inefficient coupling of the light source with the capillary limit the further improvement of UV-vis absorption detection in CE. The goals of this research have been to show the utility of laser-based absorption detection. The approaches involve: on-column double-beam laser absorption detection and its application to the detection of small ions and proteins, and absorption detection with the bubble-shaped flow cell.

  16. Nonaqueous Capillary Electrophoresis Mass Spectrometry.

    Science.gov (United States)

    Klampfl, Christian W; Himmelsbach, Markus

    2016-01-01

    The term nonaqueous capillary electrophoresis (NACE) commonly refers to capillary electrophoresis with purely nonaqueous background electrolytes (BGE). Main advantages of NACE are the possibility to analyze substances with very low solubility in aqueous media as well as separation selectivity that can be quite different in organic solvents (compared to water)-a property that can be employed for manipulation of separation selectivities. Mass spectrometry (MS) has become more and more popular as a detector in CE a fact that applies also for NACE. In the present chapter, the development of NACE-MS since 2004 is reviewed. Relevant parameters like composition of BGE and its influence on separation and detection in NACE as well as sheath liquid for NACE-MS are discussed. Finally, an overview of the papers published in the field of NACE-MS between 2004 and 2014 is given. Applications are grouped according to the field (analysis of natural products, biomedical analysis, food analysis, analysis of industrial products, and fundamental investigations). PMID:27645734

  17. Analysis of Small Ions with Capillary Electrophoresis.

    Science.gov (United States)

    Aulakh, Jatinder Singh; Kaur, Ramandeep; Malik, Ashok Kumar

    2016-01-01

    Small inorganic ions are easily separated through capillary electrophoresis because they have a high charge-to-mass ratio and suffer little from some of the undesired phenomenon affecting higher molecular weight species like adsorption to the capillary wall, decomposition, and precipitation. This chapter is focused on the analysis of small ions other than metal ions using capillary electrophoresis. Methods are described for the determination of ions of nitrogen, phosphorus, sulfur, fluorine, chlorine, bromine, and iodine. PMID:27645739

  18. Atomic Force Controlled Capillary Electrophoresis

    Science.gov (United States)

    Lewis, Aaron; Yeshua, Talia; Palchan, Mila; Lovsky, Yulia; Taha, Hesham

    2010-03-01

    Lithography based on scanning probe microscopic techniques has considerable potential for accurate & localized deposition of material on the nanometer scale. Controlled deposition of metallic features with high purity and spatial accuracy is of great interest for circuit edit applications in the semiconductor industry, for plasmonics & nanophotonics and for basic research in surface enhanced Raman scattering & nanobiophysics. Within the context of metal deposition we will review the development of fountain pen nanochemistry and its most recent emulation Atomic Force Controlled Capillary Electrophoresis (ACCE). Using this latter development we will demonstrate achievement of unprecedented control of nanoparticle deposition using a three-electrode geometry. Three electrodes are attached: one on the outside of a metal coated glass probe, one on the inside of a hollow probe in a solution containing Au nanoparticles in the capillary, and a third on the surface where the writing takes place. The three electrodes provide electrical pulses for accurate control of deposition and retraction of the liquid from the surface overcoming the lack of control seen in both dip pen lithography & fountain pen nanochemistry when the tip contacts the surface. With this development, we demonstrate depositing a single 1.3 nm Au nanoparticle onto surfaces such as semiconductors.

  19. Capillary electrophoresis - electrospray ionization mass spectrometry in small diameter capillaries

    Energy Technology Data Exchange (ETDEWEB)

    Wahl, J.H.; Goodlett, D.R.; Udseth, H.R.; Smith, R.D.

    1992-06-01

    Methods (such as small inner diameter capillaries) are being explored to increase analyte sensitivity in capillary electrophoresis- electrospray ionization/mass spectroscopy(CE-ESI/MS). Results are reported for melittin in a protein mixture, with 10 to 100 {mu}m ID capillaries; and for a mixture of aprotinin, cytochrome c, myoglobin, and carbonic anhydrase, with 5 to 50 {mu}m ID capillaries. It is shown that an increase in solute sensitivity occurs when small ID capillaries ({lt} 20 {mu}m) are used in CE-ESI/MS for both a peptide and a protein mixture. 3 figs. (DLC)

  20. A New Conductivity Detector for Capillary Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    A new conductivity detector for capillary electrophoresis consisting of an electrochemical cell and a conductive meter was developed. In the cell, the microelectrode and capillary were inserted through the cell wall and fixed by screws and sealing ring, the ends of microelectrode and capillary were located by a guide with two cross holes. LOD for K+ was 1.5×10-5 mol/L.

  1. Simultaneous determination of rare earth elements in ore and anti-corrosion coating samples using a portable capillary electrophoresis instrument with contactless conductivity detection.

    Science.gov (United States)

    Nguyen, Thi Anh Huong; Nguyen, Van Ri; Le, Duc Dung; Nguyen, Thi Thanh Binh; Cao, Van Hoang; Nguyen, Thi Kim Dung; Sáiz, Jorge; Hauser, Peter C; Mai, Thanh Duc

    2016-07-29

    The employment of an in-house-made capillary electrophoresis (CE) instrument with capacitively coupled contactless conductivity detection (C(4)D) as a simple and inexpensive solution for simultaneous determination of many rare earth elements (REEs) in ore samples from Vietnam, as well as in anti-corrosion coating samples is reported. 14 REEs (La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb and Lu) were determined using an electrolyte composed of 20mM arginine and 10mM α-hydroxyisobutyric acid adjusted to pH 4.2 with acetic acid. The best detection limit achieved was 0.24mg/L using the developed CE-C(4)D method. Good agreement between results from CE-C(4)D and the confirmation method (ICP-MS) was achieved, with a coefficient of determination (r(2)) for the two pairs of data of 0.998. PMID:27363736

  2. In-house-made capillary electrophoresis instruments coupled with contactless conductivity detection as a simple and inexpensive solution for water analysis: a case study in Vietnam.

    Science.gov (United States)

    Duong, Hong Anh; Le, Minh Duc; Nguyen, Kim Diem Mai; Hauser, Peter C; Pham, Hung Viet; Mai, Thanh Duc

    2015-11-01

    A simple and inexpensive method for the determination of various ionic species in different water matrices is discussed in this study. The approach is based on the employment of in-house-made capillary electrophoresis (CE) instruments with capacitively coupled contactless conductivity detection (C(4)D), which can be realized even when only a modest financial budget and limited expertise are available. Advantageous features and considerations of these instruments are detailed following their pilot deployment in Vietnam. Different categories of ionic species, namely major inorganic cations (K(+), Na(+), Ca(2+), Mg(2+), and NH4(+)) and major inorganic anions (Cl(-), NO3(-), NO2(-), SO4(2-), and phosphate), in different water matrices in Vietnam were determined using these in-house fabricated instruments. Inorganic trivalent arsenic (As(iii)), which is the most abundant form of arsenic in reducing groundwater, was determined by CE-C(4)D. The effect of some interfering ions in groundwater on the analytical performance was investigated and is highlighted. The results from in-house-made CE-C(4)D-instruments were cross-checked with those obtained using the standard methods (AAS, AES, UV and IC), with correlation coefficients r(2) ≥ 0.9 and deviations from the referenced results less than 15%.

  3. Microfluidic chip-capillary electrophoresis devices

    CERN Document Server

    Fung, Ying Sing; Du, Fuying; Guo, Wenpeng; Ma, Tongmei; Nie, Zhou; Sun, Hui; Wu, Ruige; Zhao, Wenfeng

    2015-01-01

    Capillary electrophoresis (CE) and microfluidic chip (MC) devices are relatively mature technologies, but this book demonstrates how they can be integrated into a single, revolutionary device that can provide on-site analysis of samples when laboratory services are unavailable. By introducing the combination of CE and MC technology, Microfluidic Chip-Capillary Electrophoresis Devices broadens the scope of chemical analysis, particularly in the biomedical, food, and environmental sciences.The book gives an overview of the development of MC and CE technology as well as technology that now allows

  4. Selectivity and detection in capillary electrophoresis

    OpenAIRE

    Khaled, Maha Yehia

    1994-01-01

    This work is a contribution to the minimization of some of the selectivity and detection limitations in capillary electrophoresis. A more practical design of an electrochemical detector is introduced with simultaneous on-line UV detection (1), for the selective detection of a number of pungent and neurological compounds, the piperines and the capsacinoids. Commercially available microelectrodes together with large 25 μm id fused silica capillary columns are used for the fir...

  5. Planetary In Situ Capillary Electrophoresis System (PISCES)

    Science.gov (United States)

    Willis, P. A.; Stockton, A. M.; Mora, M. F.; Cable, M. L.; Bramall, N. E.; Jensen, E. C.; Jiao, H.; Lynch, E.; Mathies, R. A.

    2012-10-01

    We propose to develop PISCES, a 3-kg, 2W, flight-capable microfluidic lab-on-a-chip capillary electrophoresis analyzer capable of ingesting solid, liquid, or gas samples and performing a suite of chemical analyses with parts per trillion sensitivity.

  6. Identification of inorganic improvised explosive devices by analysis of postblast residues using portable capillary electrophoresis instrumentation and indirect photometric detection with a light-emitting diode.

    Science.gov (United States)

    Hutchinson, Joseph P; Evenhuis, Christopher J; Johns, Cameron; Kazarian, Artaches A; Breadmore, Michael C; Macka, Miroslav; Hilder, Emily F; Guijt, Rosanne M; Dicinoski, Greg W; Haddad, Paul R

    2007-09-15

    A commercial portable capillary electrophoresis (CE) instrument has been used to separate inorganic anions and cations found in postblast residues from improvised explosive devices (IEDs) of the type used frequently in terrorism attacks. The purpose of this analysis was to identify the type of explosive used. The CE instrument was modified for use with an in-house miniaturized light-emitting diode (LED) detector to enable sensitive indirect photometric detection to be employed for the detection of 15 anions (acetate, benzoate, carbonate, chlorate, chloride, chlorite, cyanate, fluoride, nitrate, nitrite, perchlorate, phosphate, sulfate, thiocyanate, thiosulfate) and 12 cations (ammonium, monomethylammonium, ethylammonium, potassium, sodium, barium, strontium, magnesium, manganese, calcium, zinc, lead) as the target analytes. These ions are known to be present in postblast residues from inorganic IEDs constructed from ammonium nitrate/fuel oil mixtures, black powder, and chlorate/perchlorate/sugar mixtures. For the analysis of cations, a blue LED (470 nm) was used in conjunction with the highly absorbing cationic dye, chrysoidine (absorption maximum at 453 nm). A nonaqueous background electrolyte comprising 10 mM chrysoidine in methanol was found to give greatly improved baseline stability in comparison to aqueous electrolytes due to the increased solubility of chrysoidine and its decreased adsorption onto the capillary wall. Glacial acetic acid (0.7% v/v) was added to ensure chrysoidine was protonated and to enhance separation selectivity by means of complexation with transition metal ions. The 12 target cations were separated in less than 9.5 min with detection limits of 0.11-2.30 mg/L (calculated at a signal-to-noise ratio of 3). The anions separation system utilized a UV LED (370 nm) in conjunction with an aqueous chromate electrolyte (absorption maximum at 371 nm) consisting of 10 mM chromium(VI) oxide and 10 mM sodium chromate, buffered with 40 mM tris

  7. "Getting the best sensitivity from on-capillary fluorescence detection in capillary electrophoresis" - A tutorial.

    Science.gov (United States)

    Galievsky, Victor A; Stasheuski, Alexander S; Krylov, Sergey N

    2016-09-01

    Capillary electrophoresis with Laser-Induced Fluorescence (CE-LIF) detection is being applied to new analytical problems which challenge both the power of CE separation and the sensitivity of LIF detection. On-capillary LIF detection is much more practical than post-capillary detection in a sheath-flow cell. Therefore, commercial CE instruments utilize solely on-capillary CE-LIF detection with a Limit of Detection (LOD) in the nM range, while there are multiple applications of CE-LIF that require pM or lower LODs. This tutorial analyzes all aspects of on-capillary LIF detection in CE in an attempt to identify means for improving LOD of CE-LIF with on-capillary detection. We consider principles of signal enhancement and noise reduction, as well as relevant areas of fluorophore photochemistry and fluorescent microscopy. PMID:27543015

  8. Capillary Electrophoresis in Food and Foodomics.

    Science.gov (United States)

    Ibáñez, Clara; Acunha, Tanize; Valdés, Alberto; García-Cañas, Virginia; Cifuentes, Alejandro; Simó, Carolina

    2016-01-01

    Quality and safety assessment as well as the evaluation of other nutritional and functional properties of foods imply the use of robust, efficient, sensitive, and cost-effective analytical methodologies. Among analytical technologies used in the fields of food analysis and foodomics, capillary electrophoresis (CE) has generated great interest for the analyses of a large number of compounds due to its high separation efficiency, extremely small sample and reagent requirements, and rapid analysis. The introductory section of this chapter provides an overview of the recent applications of capillary electrophoresis (CE) in food analysis and foodomics. Relevant reviews and research articles on these topics are tabulated including papers published in the period 2011-2014. In addition, to illustrate the great capabilities of CE in foodomics the chapter describes the main experimental points to be taken into consideration for a metabolomic study of the antiproliferative effect of carnosic acid (a natural diterpene found in rosemary) against HT-29 human colon cancer cells. PMID:27645749

  9. Electrokinetic Flow and Dispersion in Capillary Electrophoresis

    Science.gov (United States)

    Ghosal, Sandip

    2006-01-01

    Electrophoretic separation of a mixture of chemical species is a fundamental technique of great usefulness in biology, health care, and forensics. In capillary electrophoresis (which has evolved from its predecessor, slab-gel electrophoresis), the sample migrates through a single microcapillary instead of through the network of pores in a gel. A fundamental design problem is to minimize dispersion in the separation direction. Molecular diffusion is inevitable and sets a theoretical limit on the best separation that can be achieved. But in practice, there are a number of effects arising out of the interplay between fluid flow, chemistry, thermal effects, and electric fields that result in enhanced dispersion. This paper reviews the subject of fluid flow in such capillary microchannels and examines the various causes of enhanced dispersion that limit the efficiency of separation.

  10. Metal Ions Analysis with Capillary Zone Electrophoresis.

    Science.gov (United States)

    Malik, Ashok Kumar; Aulakh, Jatinder Singh; Kaur, Varinder

    2016-01-01

    Capillary electrophoresis has recently attracted considerable attention as a promising analytical technique for metal ion separations. Significant advances that open new application areas for capillary electrophoresis in the analysis of metal species occurred based on various auxiliary separation principles. These are mainly due to complexation, ion pairing, solvation, and micellization interactions between metal analytes and electrolyte additives, which alter the separation selectivity in a broad range. Likewise, many separation studies for metal ions have been concentrated on the use of preelectrophoresis derivatization methodology. Approaches suitable for manipulation of selectivity for different metal species including metal cations, metal complexes, metal oxoanions, and organometallic compounds, are discussed, with special attention paid to the related electrophoretic system variables using illustrative examples. PMID:27645740

  11. Capillary Electrophoresis in the Presence of Fosfomycin

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Fosfomyein, a sodim salt of cis-(3-methyloxiranyl) phosphonic acid, was used as electrolyte in binary methanol-water media for capillary electrophoresis. The variety of electroosmotic flow with pH*,methanol concentration and ionic strength was investigated. The migration behavior of nine bases was examined under various conditions, and the separation of thymine, cytosine, 5-flurouracil, 4,6-diamino-pyrimidine, purine was accomplished.

  12. Sensitivity enhancement in direct coupling of supported liquid membrane extractions to capillary electrophoresis by means of transient isotachophoresis and large electrokinetic injections

    OpenAIRE

    Pantůčková, P. (Pavla); Kubáň, P. (Pavel); Boček, P. (Petr)

    2015-01-01

    Enhanced sensitivity for determination of basic drugs in body fluids was achieved by in-line coupling of extraction across supported liquid membrane (SLM) to large electrokinetic injection and transient isotachophoresis-capillary zone electrophoresis in commercial capillary electrophoresis instrument.

  13. CMOS absorbance detection system for capillary electrophoresis

    International Nuclear Information System (INIS)

    This paper presents a cost-effective portable photodetection system for capillary electrophoresis absorptiometry. By using a CMOS BDJ (buried double p-n junction) detector, a dual-wavelength method for absorbance measurement is implemented. This system includes associated electronics for low-noise pre-amplification and A/D conversion, followed by digital signal acquisition and processing. Two signal processing approaches are adopted to enhance the signal to noise ratio. One is variable time synchronous detection, which optimizes the sensitivity and measuring rate compared to a conventional synchronous detection technique. The other is a statistical approach based on principal component analysis, which allows optimal estimation of detected signal. This system has been designed and tested in capillary electrophoresis conditions. Its operation has been verified with performances comparable to those of a commercialized spectrophotometric system (HP-3D CE). With potential on-chip integration of associated electronics, it may be operated as an integrable detection module for microchip electrophoresis and other microanalysis systems

  14. Sheathless interface for coupling capillary electrophoresis with mass spectrometry

    Science.gov (United States)

    Wang, Chenchen; Tang, Keqi; Smith, Richard D.

    2014-06-17

    A sheathless interface for coupling capillary electrophoresis (CE) with mass spectrometry is disclosed. The sheathless interface includes a separation capillary for performing CE separation and an emitter capillary for electrospray ionization. A portion of the emitter capillary is porous or, alternatively, is coated to form an electrically conductive surface. A section of the emitter capillary is disposed within the separation capillary, forming a joint. A metal tube, containing a conductive liquid, encloses the joint.

  15. Using capillary electrophoresis to characterize polymeric particles.

    Science.gov (United States)

    Riley, Kathryn R; Liu, Sophia; Yu, Guo; Libby, Kara; Cubicciotti, Roger; Colyer, Christa L

    2016-09-01

    Capillary electrophoresis (CE) was used for the characterization of a variety of polymeric micron and sub-micron particles based on size, surface functionality, and binding properties. First, a robust capillary zone electrophoresis (CZE) method was developed for the baseline separation and quantitation of commercially available polystyrene particles with various surface modifications (including amino, carboxylate, and sulfate functional groups) and various sizes (0.2, 0.5, 1.0, and 3.0μm). The separation of DNA-templated polyacrylamide particles from untemplated particles (as used for the Ion Torrent Personal Genome Machine) was demonstrated. Finally, using the 29-base thrombin aptamer and thrombin protein as a model system, a study was undertaken to determine dissociation constants for the aptamer and protein in free solution and when the aptamer was conjugated to a particle, with the goal of better understanding how the use of solid substrates, like particles, affects selection and binding processes. Dissociation constants were determined and were found to be approximately 5-fold higher for the aptamer conjugated to a particle relative to that in free solution. PMID:27543386

  16. Capillary Electrophoresis Analysis of Cations in Water Samples: An Experiment for the Introductory Laboratory

    Science.gov (United States)

    Pursell, Christopher J.; Chandler, Bert; Bushey, Michelle M.

    2004-01-01

    Capillary electrophoresis is gradually working its way into the undergraduate laboratory curriculum. Typically, experiments utilizing this newer technology have been introduced into analytical or instrumental courses. The authors of this article have introduced an experiment into the introductory laboratory that utilizes capillary electrophoresis…

  17. Potential of capillary electrophoresis for the profiling of propolis

    NARCIS (Netherlands)

    Hilhorst, M.J; Somsen, G.W; de Jong, G.J.

    1998-01-01

    The usefulness of capillary electrophoresis (CE) with diode array detection for the profiling of Propolis, a hive product, is investigated. Water extracts of Propolis were analyzed with both capillary zone electrophoresis (CZE) at pH 7.0 and 9.3, and micellar electrokinetic chromatography (MEKC) wit

  18. Analysis of organic acids in Macedonian wines by capillary electrophoresis

    OpenAIRE

    Jancovska, Maja; Ivanova, Violeta; Gulaboski, Rubin; Belder, Detlev

    2013-01-01

    Capillary electrophoresis as a separation technique can be applied for analysis of organic acids in white and red wines, providing high resolution separation of the analytes. Organic acids such as of tartaric, malic, lactic citric and succinic acids have been analysed in many Macedonian red and white wines by capillary electrophoresis, and results have been discussed.

  19. Capillary Electrophoresis Analysis of Conventional Splicing Assays

    DEFF Research Database (Denmark)

    de Garibay, Gorka Ruiz; Acedo, Alberto; García-Casado, Zaida;

    2014-01-01

    Rare sequence variants in "high-risk" disease genes, often referred as unclassified variants (UVs), pose a serious challenge to genetic testing. However, UVs resulting in splicing alterations can be readily assessed by in vitro assays. Unfortunately, analytical and clinical interpretation...... of these assays is often challenging. Here, we explore this issue by conducting splicing assays in 31 BRCA2 genetic variants. All variants were assessed by RT-PCR followed by capillary electrophoresis and direct sequencing. If assays did not produce clear-cut outputs (Class-2 or Class-5 according to analytical...... International Agency for Research on Cancer guidelines), we performed qPCR and/or minigene assays. The latter were performed with a new splicing vector (pSAD) developed by authors of the present manuscript (patent #P201231427 CSIC). We have identified three clinically relevant Class-5 variants (c.682-2A>G, c...

  20. Nicked-sleeve interface for two-dimensional capillary electrophoresis

    OpenAIRE

    Flaherty, Ryan J.; Huge, Bonnie J.; Bruce, Spencer M.; Dada, Oluwatosin O.; Dovichi, Norman J.

    2013-01-01

    We report an improved interface for two-dimensional capillary electrophoresis. This interface is based on capillary tubing and a Plexiglas chip, both of which were milled using a micro-dicing saw. The interface was evaluated and compared to a traditional interface design for both pseudo one-dimensional and two-dimensional capillary electrophoresis. We observe less than 70% transfer efficiency for the traditional design and greater than 90% transfer efficiency with this new interface.

  1. Validation of STR typing by capillary electrophoresis.

    Science.gov (United States)

    Moretti, T R; Baumstark, A L; Defenbaugh, D A; Keys, K M; Brown, A L; Budowle, B

    2001-05-01

    With the use of capillary electrophoresis (CE), high-resolution electrophoretic separation of short tandem repeat (STR) loci can be achieved in a semiautomated fashion. Laser-induced detection of fluorescently labeled PCR products and multicolor analysis enable the rapid generation of multilocus DNA profiles. In this study, conditions for typing PCR-amplified STR loci by capillary electrophoresis were investigated using the ABI Prism 310 Genetic Analyzer (Applied Biosystems). An internal size standard was used with each run to effectively normalize mobility differences among injections. Alleles were designated by comparison to allelic ladders that were run with each sample set. Multiple runs of allelic ladders and of amplified samples demonstrate that allele sizes were reproducible, with standard deviations typically less than 0.12 bases for fragments up to 317 bases in length (largest allele analyzed) separated in a 47 cm capillary. Therefore, 99.7% of all alleles that are the same length should fall within the measurement error window of +/- 0.36 bases. Microvariants of the tetranucleotide repeats were also accurately typed by the analytical software. Alleles differing in size by one base could be resolved in two-donor DNA mixtures in which the minor component comprised > or = 5% of the total DNA. Furthermore, the quantitative data format (i.e., peak amplitude) can in some instances assist in determining individual STR profiles in mixed samples. DNA samples from previously typed cases (typed for RFLP, AmpliType PM+DQA1, and/or D1S80) were amplified using AmpFlSTR Profiler Plus and COfiler and were evaluated using the ABI Prism 310. Most samples yielded typable results. Compared with previously determined results for other loci, there were no discrepancies as to the inclusion or exclusion of suspects or victims. CE thus provides efficient separation, resolution, sensitivity and precision, and the analytical software provides reliable genotyping of STR loci. The

  2. Capillary Electrophoresis-Inductively Coupled Plasma Mass Spectrometry.

    Science.gov (United States)

    Michalke, Bernhard

    2016-01-01

    During the recent years, capillary electrophoresis (CE) has been fully established as a powerful tool in separation sciences as well as in element speciation. This road of success is based on the rapid analysis time, low sample requirements, high separation efficiency, and low operating costs of CE. Inductively coupled plasma mass spectrometry (ICP-MS) is known for superior detection and multielement capability. Consequently, the combination of both instruments is approved for analysis of complex sample types at low element concentrations which require high detection power. Also the diversity of potential applications brings CE-ICP-MS coupling into central focus of element speciation. The key to successful combination of ICP-MS as an (multi-)element selective detector for CE is the availability of a suitable and effective interface.Therefore, this chapter summarizes the most important and basic principles about coupling of capillary electrophoresis to ICP-MS. Specifically, the major requirements for interfacing are described and technical solutions are given. Such solutions include the closing of the electrical circuit from CE at the nebulization, the adoption of flow rates for efficient nebulization, the reduction of a suction flow through the capillary, caused by the nebulizer, and maintaining the high separation resolution from CE across the interface for ICP-MS detection. Additionally, detailed information is presented to determine and quantify the siphoning suction through the CE capillary by the nebulizer. Finally, two applications, namely, the manganese and selenium speciation in cerebrospinal fluid are shown as examples, providing the relevant operational parameter. PMID:27645737

  3. Fabricating PFPE Membranes for Capillary Electrophoresis

    Science.gov (United States)

    Lee, Michael C.; Willis, Peter A.; Greer, Frank; Rolland, Jason

    2009-01-01

    A process has been developed for fabricating perfluoropolyether (PFPE) membranes that contain microscopic holes of precise sizes at precise locations. The membranes are to be incorporated into laboratory-on-a-chip microfluidic devices to be used in performing capillary electrophoresis. The present process is a modified version of part of the process, described in the immediately preceding article, that includes a step in which a liquid PFPE layer is cured into solid (membrane) form by use of ultraviolet light. In the present process, one exploits the fact that by masking some locations to prevent exposure to ultraviolet light, one can prevent curing of the PFPE in those locations. The uncured PFPE can be washed away from those locations in the subsequent release and cleaning steps. Thus, holes are formed in the membrane in those locations. The most straightforward way to implement the modification is to use, during the ultraviolet-curing step, an ultraviolet photomask similar to the photomasks used in fabricating microelectronic devices. In lieu of such a photomask, one could use a mask made of any patternable ultraviolet-absorbing material (for example, an ink or a photoresist).

  4. Online comprehensive two-dimensional ion chromatography × capillary electrophoresis.

    Science.gov (United States)

    Ranjbar, Leila; Gaudry, Adam J; Breadmore, Michael C; Shellie, Robert A

    2015-09-01

    A comprehensively coupled online two-dimensional ion chromatography-capillary electrophoresis (IC × CE) system for quantitative analysis of inorganic anions and organic acids in water is introduced. The system employs an in-house built sequential injection-capillary electrophoresis instrument and a nonfocusing modulation interface comprising a tee-piece and a six-port two-position injection valve that allows comprehensive sampling of the IC effluent. High field strength (+2 kV/cm) enables rapid second-dimension separations in which each peak eluted from the first-dimension separation column is analyzed at least three times in the second dimension. The IC × CE approach has been successfully used to resolve a suite of haloacetic acids, dalapon, and common inorganic anions. Two-dimensional peak capacity for IC × CE was 498 with a peak production rate of 9 peaks/min. Linear calibration curves were obtained for all analytes from 5 to 225 ng/mL (except dibromoacetic acid (10-225 ng/mL) and tribromoacetic acid (25-225 ng/mL)). The developed approach was used to analyze a spiked tap water sample, with good measured recoveries (69-119%).

  5. Design and operation of a portable scanner for high performance microchip capillary array electrophoresis

    Science.gov (United States)

    Scherer, James R.; Liu, Peng; Mathies, Richard A.

    2010-11-01

    We have developed a compact, laser-induced fluorescence detection scanner, the multichannel capillary array electrophoresis portable scanner (McCAEPs) as a platform for electrophoretic detection and control of high-throughput, integrated microfluidic devices for genetic and other analyses. The instrument contains a confocal optical system with a rotary objective for detecting four different fluorescence signals, a pneumatic system consisting of two pressure/vacuum pumps and 28 individual addressable solenoid valves for control of on-chip microvalves and micropumps, four Polymerase Chain Reaction (PCR) temperature control systems, and four high voltage power supplies for electrophoresis. The detection limit of the instrument is ˜20 pM for on-chip capillary electrophoresis of fluorescein dyes. To demonstrate the system performance for forensic short tandem repeat (STR) analysis, two experiments were conducted: (i) electrophoretic separation and detection of STR samples on a 96-lane microfabricated capillary array electrophoresis microchip. Fully resolved PowerPlex® 16 STR profiles amplified from 1 ng of 9947A female standard DNA were successfully obtained; (ii) nine-plex STR amplification, sample injection, separation, and fluorescence detection of 100-copy 9948 male standard DNA in a single integrated PCR- capillary electrophoresis microchip. These results demonstrate that the McCAEPs can be used as a versatile control and detection instrument that operates integrated microfluidic devices for high-performance forensic human identification.

  6. Enantiomeric resolution of multiple chiral centres racemates by capillary electrophoresis.

    Science.gov (United States)

    Ali, Imran; Suhail, Mohd; Al-Othman, Zeid A; Alwarthan, Abdulrahman; Aboul-Enein, Hassan Y

    2016-05-01

    Enantiomeric resolution of multichiral centre racemates is an important area as some multichiral centre racemates are of great medicinal importance. However, enantioseparation of such types of racemates is a challenging task. Amongst many analytical techniques, capillary electrophoresis is a powerful technique and may be used to resolve such racemates. Only few papers are available describing enantiomeric resolution of such racemates. Therefore, efforts have been made to describe the enantiomeric resolution of multichiral centre racemates by capillary electrophoresis. This article discusses the importance of multichiral racemates, the need for capillary electrophoresis in enantiomeric resolution and chiral resolution of multichiral centre racemates using various chiral selectors. Further, attempts have been made to discuss the future challenges and prospects of enantiomeric resolution of multichiral racemates. The various chiral selectors used for the purpose are chiral crown ether, cyclodextrins, polysaccharides, macrocyclic glycopeptide antibiotics and ligand exchange.

  7. Optimized photonic crystal fibers supporting efficient capillary electrophoresis

    Science.gov (United States)

    Calcerrada, M.; García-Ruiz, C.; Roy, P.; Gonzalez-Herraez, M.

    2013-05-01

    In this paper we present preliminary results on the use of Photonic Crystal Fibers (PCFs) in a conventional capillary electrophoresis system to separate and detect fluorescent species. PCFs show interesting advantages over conventional capillaries for this application, including larger surface-to-volume ratio and potential for higher resolution with comparable sensitivity. Our results illustrate some of these advantages, and we point out the need for stringent tolerances in the fabrication of specific PCFs for this application.

  8. Separation and quantification of cellulases and hemicellulases by capillary electrophoresis

    DEFF Research Database (Denmark)

    Jørgensen, Henning; Kutter, Jörg Peter; Olsson, Lisbeth

    2003-01-01

    . Current methods are limited in their ability to quantify all of these enzymes when all are present simultaneously in a mixture. Five different cellulases (two cellobiohydrolases and three endoglucanases) and one hemicellulase (endoxylanase) were separated using capillary electrophoresis (CE) in a fused...... silica capillary at pH values close to neutral. The improvement of the separation of these six proteins by the addition of alpha, omega-diaminoalkanes with chain lengths from three to seven carbon units was investigated. Dynamically coating the capillary with 1,3-diaminopropane resulted in separation of...

  9. Rapid inorganic ion analysis using quantitative microchip capillary electrophoresis

    NARCIS (Netherlands)

    Vrouwe, Elwin X.; Lüttge, Regina; Olthuis, Wouter; Berg, van den Albert

    2006-01-01

    Rapid quantitative microchip capillary electrophoresis (CE) for online monitoring of drinking water enabling inorganic ion separation in less than 15s is presented. Comparing cationic and anionic standards at different concentrations the analysis of cationic species resulted in non-linear calibratio

  10. Capillary electrophoresis-based assessment of nanobody affinity and purity

    NARCIS (Netherlands)

    Haselberg, Rob; Oliveira, Sabrina; van der Meel, Roy; Somsen, Govert W; de Jong, Gerhardus J

    2014-01-01

    Drug purity and affinity are essential attributes during development and production of therapeutic proteins. In this work, capillary electrophoresis (CE) was used to determine both the affinity and composition of the biotechnologically produced "nanobody" EGa1, the binding fragment of a heavy-chain-

  11. Study of Oxidation of Glutathione by Capillary Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    A capillary electrophoresis method for the separation and quantification of reduced glutathione (GSH) and oxidized glutathione (GSSG) was developed. A baseline separation was achieved within five minutes. The effects of time and the concentrations of hydrogen peroxide (H2O2) on the oxidation of GSH were investigated.

  12. Capillary electrophoresis application in metal speciation and complexation characterization

    Science.gov (United States)

    Capillary electrophoresis is amenable to the separation of metal ionic species and the characterization of metal-ligand interactions. This book chapter reviews and discusses three representative case studies in applications of CE technology in speciation and reactions of metal with organic molecules...

  13. An axial approach to detection in capillary electrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Taylor, J.A.

    1993-05-01

    Our approach involves on-axis illumination of the compounds inside the capillary detection region and is applied to absorbance and fluorescence detection. Absorbance measurements were made by focussing an incident laser beam into one capillary end; by using signals collected over the entire length of analyte band, this enhances the analytical path length of conventional absorbance detection 60x. This instrument offers a 15x improvement in detection limits. Three fluorescence detection experiments are discussed, all of which involve insertion of an optical fiber into capillary. The first uses a high refractive index liquid phase to obtain total internal reflectance along capillary axis, this reducing light scatter. The second uses a charge-coupled device camera for simultaneous imaging of a capillary array (this may be useful in genome sequencing, etc.). The third is a study of fluid motion inside the capillary under pressure-driven and electroosmotic flow. The thesis is divided into four parts. Figs, tabs.

  14. Contactless conductivity detector for microchip capillary electrophoresis

    Science.gov (United States)

    Pumera, Martin; Wang, Joseph; Opekar, Frantisek; Jelinek, Ivan; Feldman, Jason; Lowe, Holger; Hardt, Steffen; Svehla, D. (Principal Investigator)

    2002-01-01

    A microfabricated electrophoresis chip with an integrated contactless conductivity detection system is described. The new contactless conductivity microchip detector is based on placing two planar sensing aluminum film electrodes on the outer side of a poly(methyl methacrylate) (PMMA) microchip (without contacting the solution) and measuring the impedance of the solution in the separation channel. The contactless route obviates problems (e.g., fouling, unwanted reactions) associated with the electrode-solution contact, offers isolation of the detection system from high separation fields, does not compromise the separation efficiency, and greatly simplifies the detector fabrication. Relevant experimental variables, such as the frequency and amplitude of the applied ac voltage or the separation voltage, were examined and optimized. The detector performance was illustrated by the separation of potassium, sodium, barium, and lithium cations and the chloride, sulfate, fluoride, acetate, and phosphate anions. The response was linear (over the 20 microM-7 mM range) and reproducible (RSD = 3.4-4.9%; n = 10), with detection limits of 2.8 and 6.4 microM (for potassium and chloride, respectively). The advantages associated with the contactless conductivity detection, along with the low cost of the integrated PMMA chip/detection system, should enhance the power and scope of microfluidic analytical devices.

  15. Recent advances of ionic liquids and polymeric ionic liquids in capillary electrophoresis and capillary electrochromatography.

    Science.gov (United States)

    Tang, Sheng; Liu, Shujuan; Guo, Yong; Liu, Xia; Jiang, Shengxiang

    2014-08-29

    Ionic liquids (ILs) and polymeric ionic liquids (PILs) with unique and fascinating properties have drawn considerable interest for their use in separation science, especially in chromatographic techniques. In this article, significant contributions of ILs and PILs in the improvement of capillary electrophoresis and capillary electrochromatography are described, and a specific overview of the most relevant examples of their applications in the last five years is also given. Accordingly, some general conclusions and future perspectives in these areas are discussed.

  16. Capillary Electrophoresis-based Methodology Development for Biomolecule Analysis

    OpenAIRE

    Li, Ni

    2011-01-01

    Capillary electrophoresis (CE) is a separation tool with wide applications in biomolecule analysis. Fast and high-resolution separation requiring minute sample volumes is advantageous to study multiple components in biological samples. Flexible modes and methods can be developed. In this thesis, I focus on developing and applying novel CE methods to study multi-target nucleic acid sensing with high sensitivity (Part I) and interactions between multiple components, i.e. proteins, nanoparticles...

  17. Automation and integration of multiplexed on-line sample preparation with capillary electrophoresis for DNA sequencing

    Energy Technology Data Exchange (ETDEWEB)

    Tan, H.

    1999-03-31

    The purpose of this research is to develop a multiplexed sample processing system in conjunction with multiplexed capillary electrophoresis for high-throughput DNA sequencing. The concept from DNA template to called bases was first demonstrated with a manually operated single capillary system. Later, an automated microfluidic system with 8 channels based on the same principle was successfully constructed. The instrument automatically processes 8 templates through reaction, purification, denaturation, pre-concentration, injection, separation and detection in a parallel fashion. A multiplexed freeze/thaw switching principle and a distribution network were implemented to manage flow direction and sample transportation. Dye-labeled terminator cycle-sequencing reactions are performed in an 8-capillary array in a hot air thermal cycler. Subsequently, the sequencing ladders are directly loaded into a corresponding size-exclusion chromatographic column operated at {approximately} 60 C for purification. On-line denaturation and stacking injection for capillary electrophoresis is simultaneously accomplished at a cross assembly set at {approximately} 70 C. Not only the separation capillary array but also the reaction capillary array and purification columns can be regenerated after every run. DNA sequencing data from this system allow base calling up to 460 bases with accuracy of 98%.

  18. Demonstrating Chemical and Analytical Concepts in the Undergraduate Laboratory Using Capillary Electrophoresis and Micellar Electrokinetic Chromatography

    Science.gov (United States)

    Palmer, Christopher P.

    1999-11-01

    This paper describes instrumental analysis laboratory exercises that utilize capillary electrophoresis and micellar electrokinetic chromatography to demonstrate several analytical and chemical principles. Alkyl parabens (4-hydroxy alkyl benzoates), which are common ingredients in cosmetic formulations, are separated by capillary electrophoresis. The electrophoretic mobilities of the parabens can be explained on the basis of their relative size. 3-Hydroxy ethylbenzoate is also separated to demonstrate the effect of substituent position on the acid dissociation constant and the effect this has on electrophoretic mobility. Homologous series of alkyl benzoates and alkyl phthalates (common plasticizers) are separated by micellar electrokinetic chromatography at four surfactant concentrations. This exercise demonstrates the separation mechanism of micellar electrokinetic chromatography, the concept of chromatographic phase ratio, and the concepts of micelle formation. A photodiode array detector is used in both exercises to demonstrate the advantages and limitations of the detector and to demonstrate the effect of pH and substituent position on the spectra of the analytes.

  19. [Does bilirubin interfere with capillary electrophoresis of serum proteins?].

    Science.gov (United States)

    Hellara, Ilhem; Fekih, Ons; Triki, Sonia; Elmay, Ahlem; Neffati, Fadoua; Najjar, Mohamed Fadhel

    2014-01-01

    Capillary electrophoresis of serum proteins is a fast, reliable and simple technique, but many interference exist. The objective of our work is to study the interference of bilirubin on this technique; 70 icteric sera were analysed on Capillarys ™ (Sebia). A second electrophoresis was performed on 40 samples after bilirubin photodegradation. The bilirubin and serum proteins were determinated respectively by Jendrassik and Grof and biuret methods on Konélab 20i ™ (Thermo Electron Corporation). We found abnormal spreading of the albumin fraction of the anode side wich constitute sometimes an isolated fraction in the traditional area of pre-albumin migration. This fraction varies from 2.0 ± 2.0% (0.0 to 7.3%) or 0.98 ± 1.53 g/L (0 to 5.3 g/L) and it seems to be related to the direct bilirubin since, following overloading sera with a solution of bilirubin, no further fraction was recovered. An average decrease of bilirubin after photodegradation of 58 ± 17% (26-89%) is followed by a decrease in the same order 64 ± 38% (10-100%) of the additional fraction. Acetate cellulose electrophoresis of the same samples showed no variation. The high bilirubin levels seem modify slightly the electrophoretic profile. However the impact of the interference on the interpretation of electrophoretic trace is negligible. PMID:24492101

  20. An enhanced capillary electrophoresis method for characterizing natural organic matter.

    Science.gov (United States)

    Cottrell, Barbara A; Cheng, Wei Ran; Lam, Buuan; Cooper, William J; Simpson, Andre J

    2013-02-21

    Natural organic matter (NOM) is ubiquitous and is one of the most complex naturally occurring mixtures. NOM plays an essential role in the global carbon cycle; atmospheric and natural water photochemistry; and the long-range transport of trace compounds and contaminants. There is a dearth of separation techniques capable of resolving this highly complex mixture. To our knowledge, this is the first reported use of ultrahigh resolution counterbalance capillary electrophoresis to resolve natural organic matter. The new separation strategy uses a low pH, high concentration phosphate buffer to reduce the capillary electroosmotic flow (EOF). Changing the polarity of the electrodes reverses the EOF to counterbalance the electrophoretic mobility. Sample stacking further improves the counterbalance separation. The combination of these conditions results in an electropherogram comprised up to three hundred peaks superimposed on the characteristic "humic hump" of NOM. Fraction collection, followed by three-dimensional emission excitation spectroscopy (EEMs) and UV spectroscopy generated a distinct profile of fluorescent and UV absorbing components. This enhanced counterbalance capillary electrophoresis method is a potentially powerful technique for the characterization and separation of NOM and complex environmental mixtures in general. PMID:23289095

  1. Separation of Aminobenzoic Acids by Gold Nanoparticle modified Capillary Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    YAN,Hongtao; LI,Tuo; GUO,Yanli

    2009-01-01

    A novel method for the separation of aminobenzoic acids by capillary electrophoresis was developed.The capillary was modified with gold nanoparticles.The effect of gold nanoparticles on the resolution and selectivity of separation was investigated.The influence of separation voltage,pH and buffer concentration on the separation of aminobenzoic acids was also examined.It was found that the presence of gold nanoparticles improved the precision of the analysis and increased the separation efficiency.Under the optimized experiment conditions,aminobenzoic acids were separated and determined.Linearity was established over the concentration range 0.5-40 μg·mL-1 with correlation coefficients of 0.9978-0.9992.The detection limits (S/N = 3) were from 0.1 to 0.5 μg·mL-1.

  2. Separation and determination of some carboxylic acids by capillary electrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Sladkov, V.; Fourest, B

    2006-07-01

    Separation and determination of some organic acids, mono-carboxylic (formic and acetic), dicarboxylic (oxalic and tartaric), tricarboxylic (citric) acids and aromatic acids (phtalic, benzoic, mellitic and trimellitic), by capillary electrophoresis are reviewed. The method development parameters, such as separation and injection mode, are discussed. Special attention is paid to the comparison of different detection types (spectroscopic and electrochemical). The optimisation of the carrier electrolyte composition (choice of carrier electrolyte, effect of pH, ionic strength, electro-osmotic flow modifier) is treated. Different additives (alkali-earth and transition metal ions, cyclodextrins and alcohol), which are often used for improving organic acid separation, are also considered. (authors)

  3. A New Denoising Technique for Capillary Electrophoresis Signals

    Institute of Scientific and Technical Information of China (English)

    王瑛; 莫金垣

    2002-01-01

    Capillary electrophoresis(CE) is a powerful analytical tool in chemistry,Thus,it is valuable to solve the denoising of CE signals.A new denoising method called MWDA which emplosy Mexican Hat wavelet is presented ,It is an efficient chemometrics technique and has been applied successfully in processing CE signals ,Useful information can be extractred even from signals of S/N=1 .After denoising,the peak positions are unchanged and the relative errors of peak height are less than 3%.

  4. PNEUMATIC MICROVALVE FOR ELECTROKINETIC SAMPLE PRECONCENTRATION AND CAPILLARY ELECTROPHORESIS INJECTION

    Energy Technology Data Exchange (ETDEWEB)

    Cong, Yongzheng; Rausch, Sarah J.; Geng, Tao; Jambovane, Sachin R.; Kelly, Ryan T.

    2014-10-27

    Here we show that a closed pneumatic microvalve on a PDMS chip can serve as a semipermeable membrane under an applied potential, enabling current to pass through while blocking the passage of charged analytes. Enrichment of both anionic and cationic species has been demonstrated, and concentration factors of ~70 have been achieved in just 8 s. Once analytes are concentrated, the valve is briefly opened and the sample is hydrodynamically injected onto an integrated microchip or capillary electrophoresis (CE) column. In contrast to existing preconcentration approaches, the membrane-based method described here enables both rapid analyte concentration as well as high resolution separations.

  5. Subtracting Technique of Baselines for Capillary Electrophoresis Signals

    Institute of Scientific and Technical Information of China (English)

    WANG Ying; MO Jin-yuan; CHEN Zuan-guang; GAO Yan

    2004-01-01

    The drifting baselines of capillary electrophoresis affect the veracity of analysis greatly. This paper presents Threshold Fitting Technique(TFT) so as to subtract the baselines from the original signals and emendate the signals. In TFT, wav elet and curve fitting technique are applied synthetically, thresholds are decided by the computer automatically. Many experiments of signal processing indicate that TFT is simple for being used, there are few man-induced factors, and the results are satisfactory. TFT can be applied for noisy signals without any pre-processing.

  6. Capillary electrophoresis coupled with electrochemiluminescence for determination of cloperastine hydrochloride

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Objective To investigate the electrochemiluminescence (ECL) behavior of cloperastine hydrochloride. Methods ECL intensity of tris (2,2′-bipyridyl) rutheniumo(Ⅱ) was enhanced, the method for the determination of cloperastine hydrochloride was established using capillary electrophoresis (CE) coupled with electrochemilumolinescence (ECL) detection. Results Under the optimum conditions, ECL intensity varied linearly with cloperastine hydrochloride concentration from 7.0×10-6g/mL to 1.0×10-4g/mL. The detection l...

  7. STUDY OF CAPILLARY ELECTROPHORESIS ON MICROCHIP BASED ON MEMS

    Institute of Scientific and Technical Information of China (English)

    WangMing; LiWei; 等

    2002-01-01

    Using a standard photolithographical procedure,chenmical wet etching and thermal diffusion bonding technology,a chemical analysis device for Capillary Electrophoresis(CE) has been microfabricated on a planar glass substrate with a cross-column geometry.The channels on the microchip substrate are about 50um deep and 150um wide.By employing amino acids derived from 2,4-DiNitroFluoroBenzen(DNFB) on CE chip channels,the sample manipulating system is studied based on the principle of electrodynamics.

  8. Separation and determination of some carboxylic acids by capillary electrophoresis

    International Nuclear Information System (INIS)

    Separation and determination of some organic acids, mono-carboxylic (formic and acetic), dicarboxylic (oxalic and tartaric), tricarboxylic (citric) acids and aromatic acids (phtalic, benzoic, mellitic and trimellitic), by capillary electrophoresis are reviewed. The method development parameters, such as separation and injection mode, are discussed. Special attention is paid to the comparison of different detection types (spectroscopic and electrochemical). The optimisation of the carrier electrolyte composition (choice of carrier electrolyte, effect of pH, ionic strength, electro-osmotic flow modifier) is treated. Different additives (alkali-earth and transition metal ions, cyclodextrins and alcohol), which are often used for improving organic acid separation, are also considered. (authors)

  9. [Annual review of capillary electrophoresis technology in 2015].

    Science.gov (United States)

    Wang, Xiaoqian; Zhao, Xinying; Liu, Pinduo; Wei, Qiang; Qu, Feng

    2016-02-01

    This paper reviews the capillary electrophoresis (CE) in 2015. The literatures searched from ISI Web of Science ranged in 2015. 1. 1-2015. 12. 31 are classified and introduced based on CE-MS method, methodology research, detection and enrichment, chiral separation and basic applications of CE. Six international and two national conferences are included and the important reports are introduced briefly. In the end, the standards of CE method for the analyses of monoclonal antibodies, water, wines and food approved in China and some other countries are listed. PMID:27382715

  10. Enantiomeric Separation of Meptazinol Hydrochloride by Capillary Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    YUYun-qiu; CHENYan; LINi; QIUZhui-bai

    2004-01-01

    Aim To establish a capillary electrophoresis method for enantiomerie separation of meptazinol hydrochloride. Methods The separation conditions such as cyclodextrin(CD)type, buffer pH, concentration of 2,3,6-O-triInethyl-β-cyclodextrin and organic additives were optimized. An optimum concentration was 30 mmol·L-1 phosphate (pH 7.02)with 10% (W/V) TM-β-CD and 2% acetonitrile. Results Basehne resolution of the enantiomer was readily achieved using 2,3,6-O-trimethyl-β-cyclodextrin. Conclusion This is a convenient method for fast enantiomeric resolution of meptazinol hydrochloride.

  11. Probing Antigen-Antibody Interaction Using Fluorescence Coupled Capillary Electrophoresis

    OpenAIRE

    Pengju Jiang; Jiang Xia; Jingyan Li; Cheli Wang; Yue Zhang; Lin Qiu; Jianhao Wang

    2013-01-01

    In this report, the use of fluorescence detection coupled capillary electrophoresis (CE-FL) allowed us to fully characterize the antigen-antibody interaction. CE-FL allowed separation of unbound quantum dots (QDs) and ligand bound QDs and also revealed an ordered assembly of biomolecules on QDs. Further, we observed FRET from QDs donor to DyLight acceptor, which were covalently conjugated with human IgG and goat anti-human IgG, respectively. The immunocomplex was formed and the mutual affinit...

  12. Miniaturized movable contactless conductivity detection cell for capillary electrophoresis.

    Science.gov (United States)

    Macka, Miroslav; Hutchinson, Joseph; Zemann, Andreas; Shusheng, Zhang; Haddad, Paul R

    2003-06-01

    A miniaturized capacitively coupled contactless conductivity detector (mini-C(4)D) cell has been designed which is small enough to allow it to slide along the effective capillary length inside the capillary cassette of an Agilent capiillary electrophoresis system (CE) (or other CE brand of similar construction), including the possibility of positioning it close to the point of optical detection (4 cm), or even putting two such detector cells in one cassette. The cell was tested and the performance characteristics (noise, sensitivity, and peak width) were compared with those obtained with the previously used large C(4)D cell. No significant differences were observed. The mini-C(4)D was used in simultaneous separations of common cations and anions where its advantage over a larger C(4)D cell is the ability to vary the point of detection with the mini-C(4)D cell continuously at any point along the capillary length, so that the optimum apparent selectivity can be chosen. Other applications include providing a convenient second point of detection in addition to photometric detection, such as to measure accurately the linear velocity of a zone, or to allow placement of two mini-C(4)D cells in one capillary cassette simultaneously. PMID:12858387

  13. Capacitively coupled contactless conductivity detection and sequential injection analysis in capillary electrophoresis and capillary electro-chromatography

    OpenAIRE

    Mai, Thanh Duc

    2011-01-01

    This thesis focuses on the applications of capacitively coupled contactless conductivity detection (C4D) in capillary electrophoresis (CE) hybridized with high-performance liquid chromatography (HPLC), i.e. in capillary electrochromatography and pressure-assisted capillary electrophoresis, as well as on the development and applications of an extension of CE-C4D with sequential injection analysis (SIA). At first, the in-house built C4D was used for electro-chromatographic determinations of...

  14. Mecanismos de Separação em Eletroforese Capilar Separation Mechanisms in Capillary Electrophoresis

    OpenAIRE

    Tavares, Marina F. M.

    1997-01-01

    Since its inception in the 80's, capillary electrophoresis has matured into a well established technique for the separation and analysis of complex samples. One of its strongest aspects is the ability to handle materials from a diversity of chemical classes, ranging from few to millions of Daltons. This is only possible because several modes of electrophoresis can be performed in a single capillary format. In this work, relevant aspects of capillary zone electrophoresis in its three modes (fr...

  15. Direct coupling of supported liquid membranes to capillary electrophoresis for analysis of complex samples: A tutorial

    OpenAIRE

    Kubáň, P. (Pavel); Boček, P. (Petr)

    2013-01-01

    This tutorial provides an overview of direct coupling of extraction techniques based on supported liquid membranes to capillary electrophoresis for treatment and subsequent analysis of complex samples.

  16. Applications of on-line weak affinity interactions in free solution capillary electrophoresis

    DEFF Research Database (Denmark)

    Heegaard, Niels H H; Nissen, Mogens H; Chen, David D Y

    2002-01-01

    The impressive selectivity offered by capillary electrophoresis can in some cases be further increased when ligands or additives that engage in weak affinity interactions with one or more of the separated analytes are added to the electrophoresis buffer. This on-line affinity capillary electropho......The impressive selectivity offered by capillary electrophoresis can in some cases be further increased when ligands or additives that engage in weak affinity interactions with one or more of the separated analytes are added to the electrophoresis buffer. This on-line affinity capillary...

  17. An air-pressure-free elastomeric valve for integrated nucleic acid analysis by capillary electrophoresis

    International Nuclear Information System (INIS)

    We present a new elastomeric valve for integrated nucleic acid analysis by capillary electrophoresis. The valve functions include metering to capture a designated volume of biological sample into a polymerase chain reaction (PCR) chamber, sealing to preserve the sample during PCR cycling, and transfer of the PCR-products and on-chip formamide post-processing for the analysis of DNA fragments by capillary gel electrophoresis. This new valve differs from prior art polydimethylsiloxane (PDMS) valves in that the valve is not actuated externally by air-pressure or vacuum so that it simplifies a DNA analysis system by eliminating the need for an air-pressure or vacuum source, and off-cartridge solenoid valves, control circuit boards and software. Instead, the new valve is actuated by a thermal cycling peltier assembly integrated within the hardware instrument that tightly comes in contact with a microfluidic cartridge for thermal activation during PCR, so that it spontaneously closes the valve without an additional actuator system. The valve has bumps in the designated locations so that it has a self-alignment that does not require precise alignment of a valve actuator. Moreover, the thickness of the new valve is around 600 μm with an additional bump height of 400 μm so that it is easy to handle and very feasible to fabricate by injection molding compared to other PDMS valves whose thicknesses are around 30–100 μm. The new valve provided over 95% of metering performance in filling the fixed volume of the PCR chamber, preserved over 97% of the sample volume during PCR, and showed very comparable capillary electrophoresis peak heights to the benchtop assay tube controls with very consistent transfer volume of the PCR-product and on-chip formamide. The new valve can perform a core function for integrated nucleic acid analysis by capillary electrophoresis. (paper)

  18. An air-pressure-free elastomeric valve for integrated nucleic acid analysis by capillary electrophoresis

    Science.gov (United States)

    Jung, Wooseok; Barrett, Matthew; Brooks, Carla; Rivera, Andrew; Birdsell, Dawn N.; Wagner, David M.; Zenhausern, Frederic

    2015-12-01

    We present a new elastomeric valve for integrated nucleic acid analysis by capillary electrophoresis. The valve functions include metering to capture a designated volume of biological sample into a polymerase chain reaction (PCR) chamber, sealing to preserve the sample during PCR cycling, and transfer of the PCR-products and on-chip formamide post-processing for the analysis of DNA fragments by capillary gel electrophoresis. This new valve differs from prior art polydimethylsiloxane (PDMS) valves in that the valve is not actuated externally by air-pressure or vacuum so that it simplifies a DNA analysis system by eliminating the need for an air-pressure or vacuum source, and off-cartridge solenoid valves, control circuit boards and software. Instead, the new valve is actuated by a thermal cycling peltier assembly integrated within the hardware instrument that tightly comes in contact with a microfluidic cartridge for thermal activation during PCR, so that it spontaneously closes the valve without an additional actuator system. The valve has bumps in the designated locations so that it has a self-alignment that does not require precise alignment of a valve actuator. Moreover, the thickness of the new valve is around 600 μm with an additional bump height of 400 μm so that it is easy to handle and very feasible to fabricate by injection molding compared to other PDMS valves whose thicknesses are around 30-100 μm. The new valve provided over 95% of metering performance in filling the fixed volume of the PCR chamber, preserved over 97% of the sample volume during PCR, and showed very comparable capillary electrophoresis peak heights to the benchtop assay tube controls with very consistent transfer volume of the PCR-product and on-chip formamide. The new valve can perform a core function for integrated nucleic acid analysis by capillary electrophoresis.

  19. In-capillary derivatization and capillary electrophoresis separation of amino acid neurotransmitters from brain microdialysis samples.

    Science.gov (United States)

    Denoroy, Luc; Parrot, Sandrine; Renaud, Louis; Renaud, Bernard; Zimmer, Luc

    2008-09-26

    A new in-capillary derivatization method with naphtalene-2,3-dicarboxyaldehyde (NDA)/CN(-) has been developed for capillary electrophoresis with laser-induced fluorescence detection of brain microdialysate amino acids. Samples are sandwiched between two plugs of reagent mixture at the capillary inlet and subsequently separated. Highest derivatization yields are obtained by using a reagent to sample plug length ratio equal to 4, performing a first electrophoretic mixing followed by a zero potential amplification step before applying the separation voltage and using a NaCN to NDA concentration ratio equal to 1. This new single-step methodology allows the analysis of amino acid neurotransmitters in rat brain microdialysis samples.

  20. Capillary Zone Electrophoresis-Mass Spectrometry of Intact Proteins.

    Science.gov (United States)

    Domínguez-Vega, Elena; Haselberg, Rob; Somsen, Govert W

    2016-01-01

    Capillary electrophoresis (CE) coupled with mass spectrometry (MS) has proven to be a powerful analytical tool for the characterization of intact proteins. It combines the high separation efficiency, short analysis time, and versatility of CE with the mass selectivity and sensitivity offered by MS detection. This chapter focuses on important practical considerations when applying CE-MS for the analysis of intact proteins. Technological aspects with respect to the use of CE-MS interfaces and application of noncovalent capillary coatings preventing protein adsorption are treated. Critical factors for successful protein analysis are discussed and four typical CE-MS systems are described demonstrating the characterization of different types of intact proteins by CE-MS. These methodologies comprise the use of sheath-liquid and sheathless CE-MS interfaces, and various types of noncovalent capillary coatings allowing efficient and reproducible protein separations. The discussion includes the analysis of lysozyme-drug conjugates and the therapeutic proteins human growth hormone, human interferon-β-1a, and human erythropoietin. PMID:27473479

  1. Analysis of roller pen inks by capillary zone electrophoresis

    Institute of Scientific and Technical Information of China (English)

    ZHAO Pengcheng; WANG Yanji; XU Yuanyuan; YAO Lijuan

    2007-01-01

    The analysis of roller pen inks has become more and more important in fraudulent document examination because of the extensive use of roller pens in financial documents.Capillary electrophoresis with powerful resolution was applied for the analysis of roller pen inks.The experiment focused on the optimization of the separation of the extract from commercially available roller pen entries.A better separation electropherogram was obtained when a 20 mM borate buffer at pH 8.5 and a fused silica capillary with an inner diameter of 100 μm with a total length of 47 (40 cm to the detector window)were used.Five inks from roller pens of different manufacturers and countries were analyzed,and their electropherograms showed that most patterns are distinctly different from each other.Capillary with inner diameter of 100 μm increased the intensity of determination;therefore,color dyes were identified in the visible range and were able to provide more information for comparing types of roller pen inks.

  2. Analytical characterization of wine and its precursors by capillary electrophoresis.

    Science.gov (United States)

    Gomez, Federico J V; Monasterio, Romina P; Vargas, Verónica Carolina Soto; Silva, María F

    2012-08-01

    The accurate determination of marker chemical species in grape, musts, and wines presents a unique analytical challenge with high impact on diverse areas of knowledge such as health, plant physiology, and economy. Capillary electromigration techniques have emerged as a powerful tool, allowing the separation and identification of highly polar compounds that cannot be easily separated by traditional HPLC methods, providing complementary information and permitting the simultaneous analysis of analytes with different nature in a single run. The main advantage of CE over traditional methods for wine analysis is that in most cases samples require no treatment other than filtration. The purpose of this article is to present a revision on capillary electromigration methods applied to the analysis of wine and its precursors over the last decade. The current state of the art of the topic is evaluated, with special emphasis on the natural compounds that have allowed wine to be considered as a functional food. The most representative revised compounds are phenolic compounds, amino acids, proteins, elemental species, mycotoxins, and organic acids. Finally, a discussion on future trends of the role of capillary electrophoresis in the field of analytical characterization of wines for routine analysis, wine classification, as well as multidisciplinary aspects of the so-called "from soil to glass" chain is presented.

  3. Carbon Fiber-gold/mercury Dual-electrode Detection for Capillary Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    A carbon fiber-gold/mercury dual-electrode for capillary electrophoresis is constructed. Cysteine, glutathione, ascorbic acid and uric acid can be detected simultaneously and selectively at the dual-electrode, respectively. The capillary electrophoresis / dual-electrode detection system has been used to determine these compounds in human blood samples.

  4. Determination of Enantiomeric Excess of Glutamic Acids by Lab-made Capillary Array Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    Jun WANG; Kai Ying LIU; Li WANG; Ji Ling BAI

    2006-01-01

    Simulated enantiomeric excess of glutamic acid was determined by a lab-made sixteen-channel capillary array electrophoresis with confocal fluorescent rotary scanner. The experimental results indicated that the capillary array electrophoresis method can accurately determine the enantiomeric excess of glutamic acid and can be used for high-throughput screening system for combinatorial asymmetric catalysis.

  5. Cyclodextrins in capillary electrophoresis: recent developments and new trends.

    Science.gov (United States)

    Escuder-Gilabert, L; Martín-Biosca, Y; Medina-Hernández, M J; Sagrado, S

    2014-08-29

    Despite the fact that extensive research in the field of separations by capillary electrophoresis (CE) has been carried out and many reviews have been published in the last years, a specific review on the use and future potential of cyclodextrins (CDs) in CE is not available. This review focuses the attention in the CD-CE topic over the January 2013-February 2014 period (not covered by previous more general CE-reviews). Recent contributions (reviews and research articles) including practical uses (e.g. solute-CD binding constant estimation and further potentials; 19% of publications), developments and applications (mainly chiral and achiral analysis; 38 and 24% of publications, respectively) are summarized in nine comprehensive tables and are commented. Statistics and predictions related to the CD-CE publications are highlighted in order to infer the current and expected research interests. Finally, trends and initiatives on CD-CE attending to real needs or practical criteria are outlined.

  6. Separation of enantiomers by capillary electrophoresis using pentosan polysulfate.

    Science.gov (United States)

    Wang, X; Lee, J T; Armstrong, D W

    1999-01-01

    Pentosan polysulfate, a semisynthetic polysaccharide, was employed as a chiral run buffer additive in capillary electrophoresis. Twenty-eight racemic analytes were resolved. The separations were successful only at low pH when the analytes were significantly protonated. This suggests that ionic interactions were the dominant associative interactions between the anionic pentosan polysulfate and the positively charged analytes. Compared to other linear, carbohydrate-based chiral selectors (i.e., chondroitin sulfates, heparin and dextran sulfate) pentosan polysulfate has some characteristics common of anionic polysaccharides; yet it has several differences in its structure and properties which account for its unusual enantioselectivity. The effects of pH, concentration of phosphate buffer, concentration of pentosan polysulfate and the type and concentration of organic modifier on the enantiomeric separations were investigated. The optimization of these separations were dependent on the nature of the analytes and could be achieved by the proper choice of experimental conditions.

  7. The new approach of standardization of capillary electrophoresis

    Institute of Scientific and Technical Information of China (English)

    LI; Hua; WANG; Kang; JOSEF; Havel

    2005-01-01

    In this paper, we develope the new standardization methods to eliminate the influence in capillary electrophoresis (CE). The markers were used to determine the basis position and then correct the data of sample by the migration time of standard sample, and make the migration time of samples consistent with the standard sample by the criterion of the marker. The problem of time transition was corrected in this way. Then according to the peak height or peak area of the marker in the sample (peak height was used here) compared with the standard sample, the sample data was zoomed appropriately. The absorbance error was made to be correct.The wavelet de-noise method was also used to make the data smooth and get a good baseline.

  8. Separation of ions in acidic solution by capillary electrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Thornton, M.

    1997-10-08

    Capillary electrophoresis (CE) is an effective method for separating ionic species according to differences in their electrophoretic mobilities. CE separations of amino acids by direct detection are difficult due to their similar electrophoretic mobilities and low absorbances. However, native amino acids can be separated by CE as cations at a low pH by adding an alkanesulfonic acid to the electrolyte carrier which imparts selectivity to the system. Derivatization is unnecessary when direct UV detection is used at 185 nm. Simultaneous speciation of metal cations such as vanadium (IV) and vanadium (V) can easily be performed without complexation prior to analysis. An indirect UV detection scheme for acidic conditions was also developed using guanidine as the background carrier electrolyte (BCE) for the indirect detection of metal cations. Three chapters have been removed for separate processing. This report contains introductory material, references, and general conclusions. 80 refs.

  9. Microchip capillary electrophoresis based electroanalysis of triazine herbicides.

    Science.gov (United States)

    Islam, Kamrul; Chand, Rohit; Han, Dawoon; Kim, Yong-Sang

    2015-01-01

    The number of pesticides used in agriculture is increasing steadily, leading to contamination of soil and drinking water. Herein, we present a microfluidic platform to detect the extent of contamination in soil samples. A microchip capillary electrophoresis system with in-channel electrodes was fabricated for label-free electroanalytical detection of triazine herbicides. The sample mixture contained three representative triazines: simazine, atrazine and ametryn. The electropherogram for each individual injection of simazine, atrazine and ametryn showed peaks at 58, 66 and 72 s whereas a mixture of them showed distinct peaks at 59, 67 and 71 s respectively. The technique as such may prove to be a useful qualitative and quantitative tool for the similar environmental pollutants.

  10. Recent developments in electrochemical detection for microchip capillary electrophoresis.

    Science.gov (United States)

    Vandaveer, Walter R; Pasas-Farmer, Stephanie A; Fischer, David J; Frankenfeld, Celeste N; Lunte, Susan M

    2004-11-01

    Significant progress in the development of miniaturized microfluidic systems has occurred since their inception over a decade ago. This is primarily due to the numerous advantages of microchip analysis, including the ability to analyze minute samples, speed of analysis, reduced cost and waste, and portability. This review focuses on recent developments in integrating electrochemical (EC) detection with microchip capillary electrophoresis (CE). These detection modes include amperometry, conductimetry, and potentiometry. EC detection is ideal for use with microchip CE systems because it can be easily miniaturized with no diminution in analytical performance. Advances in microchip format, electrode material and design, decoupling of the detector from the separation field, and integration of sample preparation, separation, and detection on-chip are discussed. Microchip CEEC applications for enzyme/immunoassays, clinical and environmental assays, as well as the detection of neurotransmitters are also described.

  11. Probing Antigen-Antibody Interaction Using Fluorescence Coupled Capillary Electrophoresis

    Directory of Open Access Journals (Sweden)

    Pengju Jiang

    2013-09-01

    Full Text Available In this report, the use of fluorescence detection coupled capillary electrophoresis (CE-FL allowed us to fully characterize the antigen-antibody interaction. CE-FL allowed separation of unbound quantum dots (QDs and ligand bound QDs and also revealed an ordered assembly of biomolecules on QDs. Further, we observed FRET from QDs donor to DyLight acceptor, which were covalently conjugated with human IgG and goat anti-human IgG, respectively. The immunocomplex was formed and the mutual affinity of the antigen and antibody brought QDs and DyLight close enough to allow FRET to occur. This novel CE-based technique can be easily extended to other FRET systems based on QDs and may have potential application in the detection of antibodies.

  12. Applications of capillary electrophoresis in DNA mutation analysis of genetic disorders.

    OpenAIRE

    Le, H; Fung, D.; Trent, R.J.

    1997-01-01

    AIM: To facilitate DNA mutation analysis by use of capillary electrophoresis. METHODS: The usefulness and applications of capillary electrophoresis in DNA fragment sizing and sequencing were evaluated. RESULTS: DNA mutation testing in disorders such as cystic fibrosis, Huntington disease, alpha thalassaemia, and hereditary fructose intolerance were undertaken effectively. However, sizing the (CAG)n repeat in the case of Huntington disease was a potential problem when using capillary electroph...

  13. Determination of acidity constants of enolisable compounds by capillary electrophoresis.

    Science.gov (United States)

    Mofaddel, N; Bar, N; Villemin, D; Desbène, P L

    2004-10-01

    Research on the structure-activity relationships of molecules with acidic carbon atoms led us to undertake a feasibility study on the determination of their acidity constants by capillary electrophoresis (CE). The studied molecules had diverse structures and were tetronic acid, acetylacetone, diethylmalonate, Meldrum's acid, 3-methylrhodanine, nitroacetic acid ethyl ester, pyrimidine-2,4,6-trione, 3-oxo-3-phenylpropionic acid ethyl ester, 1-phenylbutan-1,3-dione, 5,5-dimethylcyclohexan-1,3-dione and homophthalic anhydride. The p Ka range explored by CE was therefore very large (from 3 to 12) and p Ka values near 12 were evaluated by mathematical extrapolations. The analyses were carried out in CZE mode using a fused silica capillary grafted (or not) with hexadimethrine. Owing to the electrophoretic behaviour of these compounds according to the pH, their acidity constants could be evaluated and appeared in perfect agreement with the literature data obtained, a few decades ago, by means of potentiometry, spectrometry or conductimetry. The p Ka of homophthalic anhydride and 3-methylrhodanine were evaluated for the first time.

  14. Automated dual capillary electrophoresis system with hydrodynamic injection for the concurrent determination of cations and anions

    Energy Technology Data Exchange (ETDEWEB)

    Pham, Thi Thanh Thuy; Mai, Thanh Duc [University of Basel, Department of Chemistry, Spitalstrasse 51, Basel 4056 (Switzerland); Centre for Environmental Technology and Sustainable Development (CETASD), Hanoi University of Science, Nguyen Trai Street 334, Hanoi (Viet Nam); Nguyen, Thanh Dam [Centre for Environmental Technology and Sustainable Development (CETASD), Hanoi University of Science, Nguyen Trai Street 334, Hanoi (Viet Nam); Sáiz, Jorge [Department of Analytical Chemistry, Physical Chemistry and Chemical Engineering – University of Alcalá, Ctra. Madrid-Barcelona km 33.6, Alcalá de Henares, Madrid 28871 (Spain); Pham, Hung Viet, E-mail: phamhungviet@hus.edu.vn [Centre for Environmental Technology and Sustainable Development (CETASD), Hanoi University of Science, Nguyen Trai Street 334, Hanoi (Viet Nam); Hauser, Peter C., E-mail: Peter.Hauser@unibas.ch [University of Basel, Department of Chemistry, Spitalstrasse 51, Basel 4056 (Switzerland)

    2014-09-02

    Highlights: • Concurrent determination of cations and anions was carried out by electrophoretic separation. • Optimized conditions for each class of analystes was possible by using separate capillaries. • Simultaneous hydrodynamic injection was carried out. • Pneumatic actuation was used for flushing and sample handling. • The denitrification of drinking water was successfully demonstrated. - Abstract: The capillary electrophoresis instrument developed for the concurrent determination of cations and anions features two separate capillaries and individual detectors to allow independent optimization for each group of ions. The capillaries are joined in a common injector block. The sample is drawn into the injector with a small membrane pump and automated simultaneous injection into both capillaries is achieved by pressurization of the fluid with compressed air. Flushing of the injector and of the capillaries with the background electrolyte is also carried out automatically by the same means. The buffer consisted of 12 mM histidine and 2 mM 18-crown-6 adjusted to pH 4 with acetic acid and was suitable for the contactless conductivity detection employed. The system was optimized for the determination of cationic NH{sub 4}{sup +} and anionic NO{sub 3}{sup −} and NO{sub 2}{sup −}, and linear calibration curves from about 20 μM up to about 1.5 mM were obtained for these ions. In a test run over 8 h, the reproducibility for the peak areas was within ±7%. For demonstration, the instrument was successfully applied to the concurrent monitoring of the concentrations of the three ions during the biological removal of ammonium from contaminated groundwater in a sequencing batch reactor, where NO{sub 3}{sup −} and NO{sub 2}{sup −} are formed as intermediate products.

  15. Capillary zone electrophoresis and packed capillary column liquid chromatographic analysis of recombinant human interleukin-4.

    Science.gov (United States)

    Bullock, J

    1993-02-24

    Capillary zone electrophoresis (CZE) and packed capillary column liquid chromatography (micro-LC) have been applied to the analysis of the recombinant human protein interleukin-4 (rhIL-4). Separations for both the parent protein and its enzymatic digest were developed for the purpose of characterizing protein purity and identity. CZE separations of the intact protein were investigated over the pH range of 4.5 to 8.0 using uncoated fused silica capillaries. Gradient reversed-phase micro-LC was performed using 0.32 mm packed capillary columns at flow-rates of 5-6 microliters/min. Emphasis was placed on the ability of these methods to separate close structural variants and degradation products of the protein. Peptide mapping of the tryptic digest of rhIL-4 using a combination of CZE and micro-LC provided complimentary high resolution methods for establishing protein identity. Reproducible separations were achieved using sub-picomol amounts of sample. The advantages and problems encountered with these two techniques for characterizing rhIL-4 were assessed. PMID:8450025

  16. Determination of Amino Acids in Single Human Lymphocytes after On-capillary Derivatization by Capillary Zone Electrophoresis with Electrochemical Detection

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Amino acids in individual human lymphocytes were determined by capillary zone electrophoresis with electrochemical detection after on-capillary derivatization. In order to inject cells easily, a cell injector was designed. Four amino acids (serine, alanine, taurine, and glycine) in single human lymphocytes have been identified. Quantitation has been accomplished through the use of calibration curves.

  17. Capillary electrophoresis: Imaging of electroosmotic and pressure driven flow profiles in fused silica capillaries

    Science.gov (United States)

    Williams, George O., Jr.

    1996-01-01

    This study is a continuation of the summer of 1994 NASA/ASEE Summer Faculty Fellowship Program. This effort is a portion of the ongoing work by the Biophysics Branch of the Marshall Space Flight Center. The work has focused recently on the separation of macromolecules using capillary electrophoresis (CE). Two primary goals were established for the effort this summer. First, we wanted to use capillary electrophoresis to study the electrohydrodynamics of a sample stream. Secondly, there was a need to develop a methodology for using CE for separation of DNA molecules of various sizes. In order to achieve these goals we needed to establish a procedure for detection of a sample plug under the influence of an electric field Detection of the sample with the microscope and image analysis system would be helpful in studying the electrohydrodynamics of this stream under load. Videotaping this process under the influence of an electric field in real time would also be useful. Imaging and photography of the sample/background electrolyte interface would be vital to this study. Finally, detection and imaging of electroosmotic flow and pressure driven flow must be accomplished.

  18. Determination of dioxopromethazine hydrochloride by capillary electrophoresis with electrochemiluminescence detection

    International Nuclear Information System (INIS)

    The paper presents a rapid method for the determination of dioxopromethazine hydrochloride (DPZ), an antihistamine drug, by the capillary electrophoresis with electrochemiluminescene detection (CE-ECL) using tris(2,2'-bipyridyl)ruthenium(II) (Ru(bpy)32+) reagent. This CE-ECL detection method has high sensitivity, good selectivity and reproducibility for DPZ analysis. Under the optimized conditions: separation capillary, 38 cm length (25 μm i.d.); sample injection, 10 s at 8 kV; separation voltage, 12.5 kV; running buffer, 20 mmol L-1 sodium phosphate of pH 6.0; detection potential, 1.15 V; 50 mmol L-1 of phosphate buffer (pH 7.14) containing 5 mmol L-1 of Ru(bpy)32+ in ECL detection cell, the detection limit of DPZ was 0.05 μmol L-1 (S/N = 3). The linear range extended from 5 to 100 μmol L-1. The linear curve obtained was Y = 181.62 + 9.28X with a correlation coefficient of 0.9970. The relative standard deviations of the ECL intensity and the migration time for six continuous injections of 5 μmol L-1 DPZ were 3.7% and 0.92%, respectively. The CE-ECL method was applied to analyze DPZ in real samples including tablets, rat serum and human urine, and satisfactory results were obtained without interference from samples matrix. The CE-ECL technique was proved to be a potential method for the detection of DPZ in clinic analysis

  19. Analysis of Soft Drinks: UV Spectrophotometry, Liquid Chromatography, and Capillary Electrophoresis

    Science.gov (United States)

    McDevitt, Valerie L.; Rodriguez, Alejandra; Williams, Kathryn R.

    1998-05-01

    Instrumental analysis students analyze commercial soft drinks in three successive laboratory experiments. First, UV multicomponent analysis is used to determine caffeine and benzoic acid in Mello YelloTM using the spectrophotometer's software and manually by the simultaneous equations method. The following week, caffeine, benzoic acid and aspartame are determined in a variety of soft drinks by reversed-phase liquid chromatography using 45% methanol/55% aqueous phosphate, pH 3.0, as the mobile phase. In the third experiment, the same samples are analyzed by capillary electrophoresis using a pH 9.4 borate buffer. Students also determine the minimum detection limits for all three compounds by both LC and CE. The experiments demonstrate the analytical use and limitations of the three instruments. The reports and prelab quizzes also stress the importance of the chemistry of the three compounds, especially the relationships of acid/base behavior and polarity to the LC and CE separations.

  20. Study on Dicarboxylic Acids in Aerosol Samples with Capillary Electrophoresis

    Directory of Open Access Journals (Sweden)

    Heidi Adler

    2014-01-01

    Full Text Available The research was performed to study the simultaneous detection of a homologous series of α, ω-dicarboxylic acids (C2–C10, oxalic, malonic, succinic, glutaric, adipic, pimelic, suberic, azelaic, and sebacic acids, with capillary electrophoresis using indirect UV detection. Good separation efficiency in 2,6-pyridinedicarboxylic acid as background electrolyte modified with myristyl trimethyl ammonium bromide was obtained. The dicarboxylic acids were ionised and separated within five minutes. For the study, authentic samples were collected onto dry cellulose membrane filters of a cascade impactor (12 stages from outdoor spring aerosols in an urban area. Hot water and ultrasonication extraction methods were used to isolate the acids from membrane filters. Due to the low concentrations of acids in the aerosols, the extracts were concentrated with solid-phase extraction (SPE before determination. The enrichment of the carboxylic acids was between 86 and 134% with sample pretreatment followed by 100-time increase by preparation of the sample to 50 μL. Inaccuracy was optimised for all the sample processing steps. The aerosols contained dicarboxylic acids C2–C10. Then, mostly they contained C2, C5, and C10. Only one sample contained succinic acid. In the study, the concentrations of the acids in aerosols were lower than 10 ng/m3.

  1. Quantification of sugars in breakfast cereals using capillary electrophoresis.

    Science.gov (United States)

    Toutounji, Michelle R; Van Leeuwen, Matthew P; Oliver, James D; Shrestha, Ashok K; Castignolles, Patrice; Gaborieau, Marianne

    2015-05-18

    About 80% of the Australian population consumes breakfast cereal (BC) at least five days a week. With high prevalence rates of obesity and other diet-related diseases, improved methods for monitoring sugar levels in breakfast cereals would be useful in nutrition research. The heterogeneity of the complex matrix of BCs can make carbohydrate analysis challenging or necessitate tedious sample preparation leading to potential sugar loss or starch degradation into sugars. A recently established, simple and robust free solution capillary electrophoresis (CE) method was used in a new application to 13 BCs (in Australia) and compared with several established methods for quantification of carbohydrates. Carbohydrates identified in BCs by CE included sucrose, maltose, glucose and fructose. The CE method is simple requiring no sample preparation or derivatization and carbohydrates are detected by direct UV detection. CE was shown to be a more robust and accurate method for measuring carbohydrates than Fehling method, DNS (3,5-dinitrosalicylic acid) assay and HPLC (high performance liquid chromatography).

  2. Separation of Purine and Its Derivatives by Capillary Zone Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    ZengBai-zhao; ZhaoFa-qiong

    2003-01-01

    The separation of a group of 17 purine and its derivatives by capillary zone electrophoresis is presented. A systematic approach was used to study the effect of pH, buffer type, organic modifiers, applied potential, sodium dodecyl sulfate (SDS) and cyclodextrins on the separation of these purine derivatives. An ideal condition was found for their separation, which was 30 mmol/L sodium borate buffer (pH 9-9.5), 10% (V/V) methanol buffer modifier and 20 kV. Under this condition, the 17 purine derivatives were baseline separated and the linear correlation coefficient for adenine,uric acid and 2-thioxanthine was 0. 99 over two orders of magnitude. The variation of peak areas was less than 4.6%(n= 5) and that of migration times was in the range of 0%-3%, while the samples were injected hydrodynamically at a height of 15 cm and an injection time of 8-10 s. In addition,alcohol, 1-propanol, 1-butanol and acetonitrile were also effective additives in the separation. However, SDS and various β-cyclodextrin (β-CDs) were found to do no good to their separation.

  3. Separation of Purine and Its Derivatives by Capillary Zone Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    Zeng Bai-zhao; Zhao Fa-qiong

    2003-01-01

    The separation of a group of 17 purine and its derivatives by capillary zone electrophoresis is presented. A systematic approach was used to study the effect of pH, buff-er type, organic modifiers, applied potential, sodium dodecyl sulfate (SDS) and cyclodextrins on the separation of these pu-rine derivatives. An ideal condition was found for their se-paration, which was 30 mmol/L sodium borate buffer (pH 9-9.5), 10% (V/V) methanol buffer modifier and 20 kV. Un-der this condition, the 17 purine derivatives were baseline separated and the linear correlation coefficient for adenine,uric acid and 2-thioxanthine was 0. 99 over two orders of magnitude. The variation of peak areas was less than 4.6 %(n=5) and that of migration times was in the range of 0%-3%, while the samples were injected hydrodynamically at a height of 15 cm and an injection time of 8-10 s. In addition,alcohol, 1-propanol, 1-butanol and acetonitrile were also ef-fective additives in the separation. However, SDS and various β-cyclodextrin (β-CDs) were found to do no good to their se-paration.

  4. Capillary electrophoresis-chemiluminescence determination of norfloxacin and prulifloxacin

    Energy Technology Data Exchange (ETDEWEB)

    Yang Zhongju; Wang Xiaoli [College of Chemistry, Beijing Normal University, Beijing 100875 (China); Qin Weidong [College of Chemistry, Beijing Normal University, Beijing 100875 (China)], E-mail: qinwd@bnu.edu.cn; Zhao Huichun [College of Chemistry, Beijing Normal University, Beijing 100875 (China)], E-mail: zhaohuichun@bnu.edu.cn

    2008-08-15

    A capillary electrophoresis (CE)-chemiluminescence (CL) method for determining norfloxacin (NFLX) and prulifloxacin (PFLX) was developed based on the enhanced CL intensity of the cerium(IV)-sulfite-fluoroquinolone (FQ) reaction sensitized by terbium(III). The separation was conducted in buffer composed of 20 mM sodium citrate, 4 mM citric acid and 10 mM sodium sulfite at pH 6.1. The CL reagent solution consisted of 2 mM cerium(IV), 4 mM terbium(III) and 1.1 mM hydrochloric acid. NFLX and PFLX were baseline separated within 11 min with detection limits (S/N = 3) of 0.057 and 0.084 {mu}g mL{sup -1}, respectively. The maximum intra- and inter-day relative standard deviations (R.S.D.s) of migration time of the analytes were less than 4.0% and 4.2%, respectively. The proposed method was applied to detect NFLX and PFLX in fortified urine sample and the results were comparable to high-performance liquid chromatography (HPLC)-UV method. Moreover, the high selectivity of the CL detection and the high-separation efficiency of CE render the method the potential of quick analyzing fluoroquinolones in real complex matrix.

  5. Capillary electrophoresis methods for microRNAs assays: A review

    International Nuclear Information System (INIS)

    Highlights: • A review of CE analysis of miRNAs. • Summary of developments and applications of CE systems in miRNA studies. • Applications and development of microchip-based CE for rapid analysis of miRNA. - Abstract: MicroRNAs (miRNAs) are short noncoding RNAs that conduct important roles in many cellular processes such as development, proliferation, differentiation, and apoptosis. In particular, circulating miRNAs have been proposed as biomarkers for cancer, diabetes, cardiovascular disease, and other illnesses. Therefore, determination of miRNA expression levels in various biofluids is important for the investigation of biological processes in health and disease and for discovering their potential as new biomarkers and drug targets. Capillary electrophoresis (CE) is emerging as a useful analytical tool for analyzing miRNA because of its simple sample preparation steps and efficient resolution of a diverse size range of compounds. In particular, CE with laser-induced fluorescence detection is a promising and relatively rapidly developing tool with the potential to provide high sensitivity and specificity in the analysis of miRNAs. This paper covers a short overview of the recent developments and applications of CE systems in miRNA studies in biological and biomedical areas

  6. Capillary electrophoresis methods for microRNAs assays: A review

    Energy Technology Data Exchange (ETDEWEB)

    Ban, Eunmi; Song, Eun Joo, E-mail: ejsong@kist.re.kr

    2014-12-10

    Highlights: • A review of CE analysis of miRNAs. • Summary of developments and applications of CE systems in miRNA studies. • Applications and development of microchip-based CE for rapid analysis of miRNA. - Abstract: MicroRNAs (miRNAs) are short noncoding RNAs that conduct important roles in many cellular processes such as development, proliferation, differentiation, and apoptosis. In particular, circulating miRNAs have been proposed as biomarkers for cancer, diabetes, cardiovascular disease, and other illnesses. Therefore, determination of miRNA expression levels in various biofluids is important for the investigation of biological processes in health and disease and for discovering their potential as new biomarkers and drug targets. Capillary electrophoresis (CE) is emerging as a useful analytical tool for analyzing miRNA because of its simple sample preparation steps and efficient resolution of a diverse size range of compounds. In particular, CE with laser-induced fluorescence detection is a promising and relatively rapidly developing tool with the potential to provide high sensitivity and specificity in the analysis of miRNAs. This paper covers a short overview of the recent developments and applications of CE systems in miRNA studies in biological and biomedical areas.

  7. Electrochemical methods in conjunction with capillary and microchip electrophoresis.

    Science.gov (United States)

    Mark, Jonas J P; Scholz, Rebekka; Matysik, Frank-Michael

    2012-12-01

    Electromigrative techniques such as capillary and microchip electrophoresis (CE and MCE) are inherently associated with various electrochemical phenomena. The electrolytic processes occurring in the buffer reservoirs have to be considered for a proper design of miniaturized electrophoretic systems and a suitable selection of buffer composition. In addition, the control of the electroosmotic flow plays a crucial role for the optimization of CE/MCE separations. Electroanalytical methods have significant importance in the field of detection in conjunction with CE/MCE. At present, amperometric detection and contactless conductivity detection are the predominating electrochemical detection methods for CE/MCE. This paper reviews the most recent trends in the field of electrochemical detection coupled to CE/MCE. The emphasis is on methodical developments and new applications that have been published over the past five years. A rather new way for the implementation of electrochemical methods into CE systems is the concept of electrochemically assisted injection which involves the electrochemical conversions of analytes during the injection step. This approach is particularly attractive in hyphenation to mass spectrometry (MS) as it widens the range of CE-MS applications. An overview of recent developments of electrochemically assisted injection coupled to CE is presented.

  8. QuShape: Rapid, accurate, and best-practices quantification of nucleic acid probing information, resolved by capillary electrophoresis

    OpenAIRE

    Karabiber, Fethullah; McGinnis, Jennifer L.; Favorov, Oleg V.; Weeks, Kevin M.

    2013-01-01

    Chemical probing of RNA and DNA structure is a widely used and highly informative approach for examining nucleic acid structure and for evaluating interactions with protein and small-molecule ligands. QuShape, described in this paper, is a platform-independent, user-friendly software package that yields quantitative and objective nucleotide reactivity information as resolved by automated high-throughput capillary electrophoresis instruments.

  9. Determination of ephedrine and pseudoephedrine by field-amplified sample injection capillary electrophoresis.

    Science.gov (United States)

    Deng, Dongli; Deng, Hao; Zhang, Lichun; Su, Yingying

    2014-04-01

    A simple and rapid capillary electrophoresis method was developed for the separation and determination of ephedrine (E) and pseudoephedrine (PE) in a buffer solution containing 80 mM of NaH2PO4 (pH 3.0), 15 mM of β-cyclodextrin and 0.3% of hydroxypropyl methylcellulose. The field-amplified sample injection (FASI) technique was applied to the online concentration of the alkaloids. With FASI in the presence of a low conductivity solvent plug (water), an approximately 1,000-fold improvement in sensitivity was achieved without any loss of separation efficiency when compared to conventional sample injection. Under these optimized conditions, a baseline separation of the two analytes was achieved within 16 min and the detection limits for E and PE were 0.7 and 0.6 µg/L, respectively. Without expensive instruments or labeling of the compounds, the limits of detection for E and PE obtained by the proposed method are comparable with (or even lower than) those obtained by capillary electrophoresis laser-induced fluorescence, liquid chromatography-tandem mass spectrometry and gas chromatography-mass spectrometry. The method was validated in terms of precision, linearity and accuracy, and successfully applied for the determination of the two alkaloids in Ephedra herbs.

  10. Determination of Size Distribution of Nano-particles by Capillary Zone Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    Yan XUE; Hai Ying YANG; Yong Tan YANG

    2005-01-01

    A new method was developed for the determination of the size distribution of nano-particles by capillary zone electrophoresis (CZE). Scattering effect of nanoparticles was studied. This method for the determination of size distribution was statistical.

  11. Monitoring Homovanillic Acid and Vanillylmandelic Acid in Human Urine by Capillary Electrophoresis with Electrochemical Detection

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    A simple, rapid and low-cost method of separation and determination of homovanillic acid and vanillylmandelic acid in human urine was developed based on capillary zone electrophoresis / amperometric detection with high sensitivity and good resolution.

  12. Trace analysis of organic ions in ice samples by capillary electrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Huber, T. [Bern Univ. (Switzerland); Schwikowski, M.; Gaeggeler, H.W. [Paul Scherrer Inst. (PSI), Villigen (Switzerland)

    1997-09-01

    Capillary electrophoresis was tested as a new analytical method for ice samples. Comparisons to ion chromatography were made concerning accuracy, detection limits, reproducibility, necessary sample volume and time consumption. (author) 1 fig., 3 refs.

  13. Capillary Electrophoresis Profiles and Fluorophore Components of Humic Acids in Nebraska Corn and Philippine Rice Soils

    Science.gov (United States)

    As humic substances represent relatively high molecular mass polyelectrolytes containing aromatic, aliphatic and heterocyclic subunits, capillary electrophoresis (CE) has become an attractive method for “finger-print” characterization of humic acids. In addition, fluorescence excitation-emission ma...

  14. Synthesis and Characterization of Water-Soluble Carboxymethyl-Cyclodextrin Polymer as Capillary Electrophoresis Chiral Selector

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    The water-soluble carboxymethyl-cyclodextrin polymer (CM-CD polymer) was synthesized and used as capillary electrophoresis chiral selector.Verrapamil and thiopentorusodium were well separated using CM-CD polymer as chiral selector.

  15. p-Hydrazinobenzenesulfonic Acid Derivatives of Carbohydrates and Their Capillary Zone Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    p-Hydrazinobenzenesulfonic acid is explored as a novel ultraviolet labeling reagent for capillary electrophoresis (CE) of mono- and disaccharides. The labeling reaction takes less than 10 minutes and introduces both of absorption and charge groups into the sugars.

  16. Recent advances in the analysis of biological particles by capillary electrophoresis

    OpenAIRE

    Kostal, Vratislav; Arriaga, Edgar A.

    2008-01-01

    This review covers research papers published in the years 2005–2007 that describe the application of capillary electrophoresis to the analysis of biological particles such as whole cells, subcellular organelles, viruses and microorganisms.

  17. ANALYSIS OF THE ENANTIOMERS OF CHIRAL PESTICIDES AND OTHER POLLUTANTS IN ENVIRONMENTAL SAMPLES BY CAPILLARY ELECTROPHORESIS

    Science.gov (United States)

    The generic method described here involves typical capillary electrophoresis (CE) techniques, with the addition of cyclodextrin chiral selectors to the electrolyte for enantiomer separation and also, in the case of neutral analytes, the further addition of a micelle forming comp...

  18. DNA sequencing with capillary electrophoresis and single cell analysis with mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Fung, N.

    1998-03-27

    Since the first demonstration of the laser in the 1960`s, lasers have found numerous applications in analytical chemistry. In this work, two different applications are described, namely, DNA sequencing with capillary gel electrophoresis and single cell analysis with mass spectrometry. Two projects are described in which high-speed DNA separations with capillary gel electrophoresis were demonstrated. In the third project, flow cytometry and mass spectrometry were coupled via a laser vaporization/ionization interface and individual mammalian cells were analyzed. First, DNA Sanger fragments were separated by capillary gel electrophoresis. A separation speed of 20 basepairs per minute was demonstrated with a mixed poly(ethylene oxide) (PEO) sieving solution. In addition, a new capillary wall treatment protocol was developed in which bare (or uncoated) capillaries can be used in DNA sequencing. Second, a temperature programming scheme was used to separate DNA Sanger fragments. Third, flow cytometry and mass spectrometry were coupled with a laser vaporization/ionization interface.

  19. Chiral Separation by Capillary Zone Electrophoresis Used Cyclodextrins and Their Derivatives as Chiral Selector

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    @@ Capillary zone electrophoresis (CZE) is a very pronising analytical technique for the optical isomer resolution of the compounds studied. The drawbacks of the techniques such as HPLC [1] were sophisticated stationary phases and/or the relatively high quantity of the chiral agent in the mobile phase, which do not exist in CZE. The capillary electrophoresis (CE) method can offer advantages on lower consumption of analyte and background electrolyte (BGE), shorter analysis time, and higher efficiencies [2-3

  20. Quantification of Carbohydrates in Grape Tissues Using Capillary Zone Electrophoresis

    Science.gov (United States)

    Zhao, Lu; Chanon, Ann M.; Chattopadhyay, Nabanita; Dami, Imed E.; Blakeslee, Joshua J.

    2016-01-01

    Soluble sugars play an important role in freezing tolerance in both herbaceous and woody plants, functioning in both the reduction of freezing-induced dehydration and the cryoprotection of cellular constituents. The quantification of soluble sugars in plant tissues is, therefore, essential in understanding freezing tolerance. While a number of analytical techniques and methods have been used to quantify sugars, most of these are expensive and time-consuming due to complex sample preparation procedures which require the derivatization of the carbohydrates being analyzed. Analysis of soluble sugars using capillary zone electrophoresis (CZE) under alkaline conditions with direct UV detection has previously been used to quantify simple sugars in fruit juices. However, it was unclear whether CZE-based methods could be successfully used to quantify the broader range of sugars present in complex plant extracts. Here, we present the development of an optimized CZE method capable of separating and quantifying mono-, di-, and tri-saccharides isolated from plant tissues. This optimized CZE method employs a column electrolyte buffer containing 130 mM NaOH, pH 13.0, creating a current of 185 μA when a separation voltage of 10 kV is employed. The optimized CZE method provides limits-of-detection (an average of 1.5 ng/μL) for individual carbohydrates comparable or superior to those obtained using gas chromatography–mass spectrometry, and allows resolution of non-structural sugars and cell wall components (structural sugars). The optimized CZE method was successfully used to quantify sugars from grape leaves and buds, and is a robust tool for the quantification of plant sugars found in vegetative and woody tissues. The increased analytical efficiency of this CZE method makes it ideal for use in high-throughput metabolomics studies designed to quantify plant sugars. PMID:27379118

  1. Capillary electrophoresis as a versatile tool for the bioanalysis of drugs - a review

    NARCIS (Netherlands)

    Boone, CM; Waterval, JCM; Lingeman, H; Ensing, K; Underberg, WJM

    1999-01-01

    This review article presents an overview of current research on the use of capillary electrophoretic techniques for the analysis of drugs in biological matrices. The principles of capillary electrophoresis and its various separation and detection modes are briefly discussed. Sample pretreatment meth

  2. An absorption detection approach for multiplexed capillary electrophoresis using a linear photodiode array.

    Science.gov (United States)

    Gong, X; Yeung, E S

    1999-11-01

    A novel absorption detection method for highly multiplexed capillary electrophoresis is presented for zone electrophoresis and for micellar electrokinetic chromatography. The approach involves the use of a linear photodiode array on which a capillary array is imaged by a camera lens. Either a tungsten lamp or a mercury lamp can be used as the light source such that all common wavelengths for absorption detection are accessible by simply interchanging narrow-band filters. Each capillary spans several diodes in the photodiode array for absorption measurements. Over 100 densely packed capillaries can be monitored by a single photodiode array element with 1024 diodes. The detection limit for rhodamine 6G for each capillary in the multiplexed array is ∼1.8 × 10(-)(8) M injected (S/N = 2). The cross-talk between adjacent capillaries is less than 0.2%. Simultaneous analysis of 96 samples is demonstrated. PMID:21662842

  3. Evaluation of The Interaction between Netropsin and Double Stranded DNA by Capillary Zone Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    Capillary zone electrophoresis (CZE) was applied to study the interaction between netropsin and a 14mer double stranded DNA (dsDNA). The binding constant of this interaction calculated from Scatchard plot was (1.07±0.10)×105 (mol/L)-1. The binding stoichiometry was 1:1. The use of polyacrylamide coated capillary showed better effect in the analysis of DNA than noncoated capillary.

  4. Modification of resolution in capillary electrophoresis for protein profiling in identification of genetic modification in foods

    OpenAIRE

    Latoszek, A.; Cifuentes, Alejandro

    2011-01-01

    The capillary electrophoresis with UV detection was employed for protein profiling in extracts from maize and soybeans. Modifications of back-ground electrolyte and coating the capillary wall with polybrene was employed in order to decrease the protein adsorption on the capillary walls. The obtained protein profiles were compared for transgenic and non-transgenic variants, showing in some cases significant changes that might be employed for identification of genetic modifications ...

  5. Capillary zone electrophoresis analysis and detection of mid-spectrum biological warfare agents. Suffield memorandum No. 1463

    Energy Technology Data Exchange (ETDEWEB)

    Boulet, C.A.

    1995-12-31

    Mid-spectrum biological warfare agents such as proteins, peptides, and toxins are often difficult to analyze and often require individually developed assay methods for detection and identification. In this regard, capillary electrophoresis is an important, emerging technique for separation and quantitation of peptides and proteins, providing separation efficiencies up to two orders of magnitude greater than high performance liquid chromatography. The technique can also analyze a broad range of compounds, has a simple instrument design which can be automated, and has low sample volume requirements. In this study, a highly efficient and reproducible capillary zone electrophoresis method was developed to separate and identify a series of nine peptides of defense interest including bradykinin, leucine enkephalin, and oxytocin. The paper demonstrates three strategies which could be used in a fully automated field detection and identification system for unknown peptides.

  6. Analysis of neuropeptides using capillary zone electrophoresis with multichannel fluorescence detection

    Science.gov (United States)

    Sweedler, Jonathan V.; Shear, Jason B.; Fishman, Harvey A.; Zare, Richard N.; Scheller, Richard H.

    1991-12-01

    Capillary zone electrophoresis is fast becoming one of the most sensitive separation schemes for sampling complex microenvironments. A unique detection scheme is developed in which a charge-coupled device (CCD) detects laser induced fluorescence from an axially illuminated electrophoresis capillary. The fluorescence from an analyte band is measured over a several centimeter section of the capillary, greatly increasing the observation time of the fluorescently tagged band. The sensitivity of the system is in the 1-8 X 10-20 mol range for derivatized amino acids and peptides. Subattomole quantities of bag cell neuropeptides collected from the giant marine mollusk Aplysia californica can be measured.

  7. Capillary electrophoresis of miniSTR markers to genotype highly degraded DNA samples.

    Science.gov (United States)

    Coble, Michael D

    2012-01-01

    The amplification of short tandem repeat (STR) markers throughout the human nuclear DNA genome are used to associate crime scene evidence to the perpetrator's profile in criminal investigations. For highly challenged or compromised materials such as stains exposed to the elements, skeletal remains from missing persons cases, or fragmented and degraded samples from mass disasters, obtaining a full STR profile may be difficult if not impossible. With the introduction of short amplicon STR or "miniSTR" typing, it is possible to obtain STR genetic information from highly challenged samples without the need to sequence the hypervariable regions of the mitochondrial DNA (mtDNA) genome. Non-Combined DNA Index System (CODIS) STR markers have been developed to obtain information beyond the core CODIS loci. This chapter will focus on the steps necessary to prepare and use one of the non-CODIS (NC) multiplexes, NC01 (Coble and Butler 2005), for analysis on capillary electrophoresis instrumentation.

  8. Using Capillary Electrophoresis to Determine the Purity of Acetylsalicylic Acid Synthesized in the Undergraduate Laboratory

    Science.gov (United States)

    Welder, Frank; Colyer, Christa L.

    2001-11-01

    Capillary electrophoresis (CE), although a powerful analytical tool, has found only limited application in undergraduate laboratory study. In an effort to expose freshman and sophomore chemistry students to this technique, thereby giving them practical instrumental experience early in their careers, we propose to use CE in the analysis of student-synthesized acetylsalicylic acid (ASA). The synthesis of ASA from salicylic acid (SA) is a routine undergraduate laboratory, although students rarely have the opportunity to test the purity of their product. The CE method described herein provides students with a method to test purity and yield of their product and to determine the effect of aging on their sample. CE can accomplish this in a short period of time, with minimal disruption to the regular laboratory curriculum. Optimized separation conditions, limits of detection, and linear range for ASA and SA are also given.

  9. Determination of anions with an on-line capillary electrophoresis method; Anionien on-line maeaeritys kapillaarielektroforeesilla - MPKT 10

    Energy Technology Data Exchange (ETDEWEB)

    Siren, H.; Saerme, T.; Kotiaho, T.; Hiissa, T.; Savolahti, P.; Komppa, V. [VTT Chemical Technology, Espoo (Finland)

    1998-12-31

    The aim of the study was to set-up an on-line capillary electrophoresis method for determination of anions in process waters of pulp and paper industry with exporting the results to the process control system of the mill. The quantification is important, since it will give information about the possible causes of precipitation. In recent years, the capillary electrophoresis (CE) due to its high separation efficiency has been shown as a method to take into consideration when analyzing chemical species ranging from small inorganic anions to different macromolecules. Many compounds are not easily detected in their native state, why analysis methods must be developed to improve their detection. Especially, small inorganic and organic anions which do not have chromophores are not sensitive enough for direct-UV detection. In such analyses the anions are mostly detected with indirect-UV technique. Capillary electrophoresis instruments are used to analyze samples in off-line, which seldom represent the situation in process. Therefore, on-line instrument technology with autoanalyzing settings will be needed in quality control. The development of a fully automatic capillary electrophoresis system is underway in co-operation with KCL (The Finnish Pulp and Paper Research Institute). In our research, we have first concentrated on the determination of sulphate in waters of paper industry. The method used for detection of sulphate is based on indirect-UV detection with CE, where the background electrolyte (BGE) is an absorbing mixture of secondary amines. The whole procedure for quantification of sulphate is performed within 15 minutes, after which a new sample is analyzed automatically. The only sample pretreatment is filtration, which is necessary before analysis. The concentrations of sulphate in process waters tested were between 300 and 800 ppm. Our tests show that a simultaneous determination of chloride, sulphate, nitrate, nitrite, sulphite, carbonate and oxalate is also

  10. Recent advances in amino acid analysis by capillary electrophoresis.

    Science.gov (United States)

    Poinsot, Véréna; Carpéné, Marie-Anne; Bouajila, Jalloul; Gavard, Pierre; Feurer, Bernard; Couderc, François

    2012-01-01

    This paper describes the most important articles that have been published on amino acid analysis using CE during the period from June 2009 to May 2011 and follows the format of the previous articles of Smith (Electrophoresis 1999, 20, 3078-3083), Prata et al. (Electrophoresis 2001, 22, 4129-4138) and Poinsot et al. (Electrophoresis 2003, 24, 4047-4062; Electrophoresis 2006, 27, 176-194; Electrophoresis 2008, 29, 207-223; Electrophoresis 2010, 31, 105-121). We present new developments in amino acid analysis with CE, which are reported describing the use of lasers or light emitting diodes for fluorescence detection, conductimetry electrochemiluminescence detectors, mass spectrometry applications, and lab-on-a-chip applications using CE. In addition, we describe articles concerning clinical studies and neurochemical applications of these techniques.

  11. Determination of methicillin-resistant and methicillin-susceptible Staphylococcus aureus bacteria in blood by capillary zone electrophoresis

    OpenAIRE

    Horká, M. (Marie); Tesařová, M. (Marie); Karásek, P. (Pavel); Růžička, F.; Holá, V.; Sittová, M.; Roth, M

    2015-01-01

    We used capillary zone electrophoresis in supercritical water-etched and modified fused silica capillaries to separate methicillin-resistant and methicillin-susceptible Staphylococcus aureus bacteria from clinical samples of whole blood.

  12. Determination of preservatives in soft drinks by capillary electrophoresis with ionic liquids as the electrolyte additives.

    Science.gov (United States)

    Sun, Bingbing; Qi, Li; Wang, Minglin

    2014-08-01

    A capillary electrophoresis method for separating preservatives with various ionic liquids as the electrolyte additives has been developed. The performances for separation of the preservatives using five ionic liquids with different anions and different substituted group numbers on imidazole ring were studied. After investigating the influence of the key parameters on the separation (the concentration of ionic liquids, pH, and the concentration of borax), it has been found that the separation efficiency could be improved obviously using the ionic liquids as the electrolyte additives and tested preservatives were baseline separated. The proposed capillary electrophoresis method exhibited favorable quantitative analysis property of the preservatives with good linearity (r(2) = 0.998), repeatability (relative standard deviations ≤ 3.3%) and high recovery (79.4-117.5%). Furthermore, this feasible and efficient capillary electrophoresis method was applied in detecting the preservatives in soft drinks, introducing a new way for assaying the preservatives in food products.

  13. An integrated multiple capillary array electrophoresis system for high-throughput DNA sequencing

    Energy Technology Data Exchange (ETDEWEB)

    Lu, X.

    1998-03-27

    A capillary array electrophoresis system was chosen to perform DNA sequencing because of several advantages such as rapid heat dissipation, multiplexing capabilities, gel matrix filling simplicity, and the mature nature of the associated manufacturing technologies. There are two major concerns for the multiple capillary systems. One concern is inter-capillary cross-talk, and the other concern is excitation and detection efficiency. Cross-talk is eliminated through proper optical coupling, good focusing and immersing capillary array into index matching fluid. A side-entry excitation scheme with orthogonal detection was established for large capillary array. Two 100 capillary array formats were used for DNA sequencing. One format is cylindrical capillary with 150 {micro}m o.d., 75 {micro}m i.d and the other format is square capillary with 300 {micro}m out edge and 75 {micro}m inner edge. This project is focused on the development of excitation and detection of DNA as well as performing DNA sequencing. The DNA injection schemes are discussed for the cases of single and bundled capillaries. An individual sampling device was designed. The base-calling was performed for a capillary from the capillary array with the accuracy of 98%.

  14. Development of fully automated quantitative capillary electrophoresis with high accuracy and repeatability.

    Science.gov (United States)

    Xu, Yuan; Ling, Bang-Zan; Zhu, Wen-Jun; Yao, Dong; Zhang, Lin; Wang, Yan; Yan, Chao

    2016-03-01

    A quantitative capillary electrophoresis (qCE) was developed by utilizing a rotary type of nano-volume injector, an autosampler, and a thermostat with cooling capacity. The accuracy and precision were greatly improved compared with conventional capillary electrophoresis. The 10 nL volume accuracy was guaranteed by the carefully designed nano-injector with an accurate internal loop. The system repeatability (precision) in terms of RSD test sample. We believe that this fully automated qCE system has the potential to be employed broadly in quality control and quality assurance in the pharmaceutical industry. PMID:26174138

  15. A New Dual-electrode and Multi-channel Electrochemical DetectionSystem for Capillary Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    Bing Yi YANG; Jin Yuan MO; Rong LAI

    2004-01-01

    A new type of dual-electrode and multi-channel electrochemical detection technology for capillary electrophoresis is described in this paper. Two detectors(the amperometric detector and the conductometric detector)or two conductometric detectors are connected to the same capillary electrophoresis system. The whole system possesses the advantages of the two electrochemical detectors including sparing time,improving the analytical speed and expanding the sample range.The working electrode and detector cell are handled easily.The system was applied to sample detection with satisfactory results.

  16. Chiral Separation by Capillary Zone Electrophoresis Used Cyclodextrins and Their Derivatives as Chiral Selector

    Institute of Scientific and Technical Information of China (English)

    HOU; JingGuo

    2001-01-01

    Capillary zone electrophoresis (CZE) is a very pronising analytical technique for the optical isomer resolution of the compounds studied. The drawbacks of the techniques such as HPLC [1] were sophisticated stationary phases and/or the relatively high quantity of the chiral agent in the mobile phase, which do not exist in CZE. The capillary electrophoresis (CE) method can offer advantages on lower consumption of analyte and background electrolyte (BGE), shorter analysis time, and higher efficiencies [2-3]  ……

  17. Development of fully automated quantitative capillary electrophoresis with high accuracy and repeatability.

    Science.gov (United States)

    Xu, Yuan; Ling, Bang-Zan; Zhu, Wen-Jun; Yao, Dong; Zhang, Lin; Wang, Yan; Yan, Chao

    2016-03-01

    A quantitative capillary electrophoresis (qCE) was developed by utilizing a rotary type of nano-volume injector, an autosampler, and a thermostat with cooling capacity. The accuracy and precision were greatly improved compared with conventional capillary electrophoresis. The 10 nL volume accuracy was guaranteed by the carefully designed nano-injector with an accurate internal loop. The system repeatability (precision) in terms of RSD automated qCE system has the potential to be employed broadly in quality control and quality assurance in the pharmaceutical industry.

  18. Assay of Histamine in Single Mast Cells by Capillary Zone Electrophoresis with Electrochemical Detection

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Capillary zone electrophoresis was employed for the analysis of histamine in single rat peritoneal mast cells using an amperometric detector. In this method, individual mast cells and then 0.02 mol/L NaOH as a lysing solution are injected into the front end of the separation capillary. A cell injector was constructed for easy injection of single cells. Histamine in single mast cells has been identified and quantified.

  19. Capillary zone electrophoresis for separation and quantitative determination of mexiletine and its main phase I metabolites.

    Science.gov (United States)

    Bruno, Claudio; Cavalluzzi, Maria Maddalena; Carocci, Alessia; Catalano, Alessia; Franchini, Carlo; Lentini, Giovanni

    2013-03-01

    The simultaneous separation and quantification of the analytes within the minimum analysis time and the maximum resolution and efficiency are the main objectives in the development of a capillary electrophoretic method for the determination of solutes. In this paper we describe a specific, sensitive and robust method, using capillary zone electrophoresis with internal standard and UV detection, for the separation and quantification of the anti-arrhythmic drug mexiletine, its main phase I metabolites, and its main nitrogenous degradation product. PMID:23826880

  20. Identification of inorganic improvised explosive devices using sequential injection capillary electrophoresis and contactless conductivity detection.

    Science.gov (United States)

    Blanco, Gustavo A; Nai, Yi H; Hilder, Emily F; Shellie, Robert A; Dicinoski, Greg W; Haddad, Paul R; Breadmore, Michael C

    2011-12-01

    A simple sequential injection capillary electrophoresis (SI-CE) instrument with capacitively coupled contactless conductivity detection (C(4)D) has been developed for the rapid separation of anions relevant to the identification of inorganic improvised explosive devices (IEDs). Four of the most common explosive tracer ions, nitrate, perchlorate, chlorate, and azide, and the most common background ions, chloride, sulfate, thiocyanate, fluoride, phosphate, and carbonate, were chosen for investigation. Using a separation electrolyte comprising 50 mM tris(hydroxymethyl)aminomethane, 50 mM cyclohexyl-2-aminoethanesulfonic acid, pH 8.9 and 0.05% poly(ethyleneimine) (PEI) in a hexadimethrine bromide (HDMB)-coated capillary it was possible to partially separate all 10 ions within 90 s. The combination of two cationic polymer additives (PEI and HDMB) was necessary to achieve adequate selectivity with a sufficiently stable electroosmotic flow (EOF), which was not possible with only one polymer. Careful optimization of variables affecting the speed of separation and injection timing allowed a further reduction of separation time to 55 s while maintaining adequate efficiency and resolution. Software control makes high sample throughput possible (60 samples/h), with very high repeatability of migration times [0.63-2.07% relative standard deviation (RSD) for 240 injections]. The separation speed does not compromise sensitivity, with limits of detection ranging from 23 to 50 μg·L(-1) for all the explosive residues considered, which is 10× lower than those achieved by indirect absorbance detection and 2× lower than those achieved by C(4)D using portable benchtop instrumentation. The combination of automation, high sample throughput, high confidence of peak identification, and low limits of detection makes this methodology ideal for the rapid identification of inorganic IED residues.

  1. Advances in capillary electrophoresis : In-line preconcentration for biomedical analysis. Impurity profiling of heparin

    NARCIS (Netherlands)

    van der Hoorn, Y.H.

    2015-01-01

    Capillary electrophoresis (CE) has shown to be highly suitable for the analysis of polar and ionogenic compounds in biomedical and pharmaceutical samples. Separation with CE is based on the charge-to-size ratio of analytes. The application of CE for bioanalysis may be hindered by its relatively low

  2. Nanometal-Oxide Sunscreen Agents by Capillary Electrophoresis-Inductively Coupled Plasma Mass Spectrometry

    Science.gov (United States)

    Capillary electrophoresis with detection by ICPMS is being explored to characterize nanomaterials in waste water treatment plant effluents. TiO2 and ZnO, being widely used as UV filters in personal care products, plastics, and paints, are of concern as they enter the environment...

  3. Capillary Electrophoresis and Fluorescence Excitation-Emission Matrix Characterization of Soil Mobile and Calcium Humates

    Science.gov (United States)

    Capillary electrophoresis (CE) and Excitation-emission matrix (EEM) fluorescence spectroscopy have been used in natural organic matter (NOM) studies. The mutual relevance of data collected from each of the two methods provides novel insight into the correlation of complex NOM fluorescence spectra to...

  4. SIMULTANEOUS DTERMINATION OF CHROMATE AND AROMATIC HYDROCARBONS IN ENVIRONMENTAL SAMPLES BY CAPILLARY ELECTROPHORESIS

    Science.gov (United States)

    An analytical method was developed to determine simultaneously, the inorganic anion CrO2-4, and organic aromatic compounds including benzoate, 2-Cl-benzoate, phenol, m-cresol and o-/p-cresol by capillary electrophoresis (CE). Chromate and the aromatics were separated in a relativ...

  5. Capillary electrophoresis of the mycotoxin zearalenone using cyclodextrin-enhanced fluorescence

    Science.gov (United States)

    Certain of the cyclodextrins are capable of significantly enhancing the native fluorescence of the estrogenic mycotoxin zearalenone (ZEN). Twenty-two cyclodextrins (CDs) were screened for their ability to enhance the fluorescence of ZEN in a capillary electrophoresis-laser induced fluorescence (CE-...

  6. Determination of the specific activities of methionine sulfoxide reductase A and B by capillary electrophoresis

    Science.gov (United States)

    A capillary electrophoresis (CE) method for the determination of methionine sulfoxide reductase A and methionine sulfoxide reductase B activities in mouse liver is described. The method is based on detection of the 4-(dimethylamino)azobenzene-4’-sulfonyl derivative of L-methionine (dabsyl Met), the ...

  7. On-Line Multichannel Raman Spectroscopic Detection System For Capillary Zone Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    An on-line multichannel Raman spectroscopic detection system for capillary electrophoresis was established by using an Ar+ laser and a cryogenically cooled ICCD. Resonant excitation Raman spectra of methyl red and methyl orange were employed to test the system. The result shows that it could yield on-line electrophoretogram and time series of Raman spectra.

  8. Study of Oxidation of Glutathione Treated with Hypochlorous Acid by Capillary Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Capillary electrophoresis (CE) method was developed for the separation and quantification of reduced glutathione (GSH), oxidized glutathione (GSSG) and glutathione sulphonic acid (GSO3H). Baseline separation was obtained within five minutes. The effects of reaction time and molar ratio of hypochlorous acid (HOCI) to GSH on the oxidation of GSH were investigated.

  9. Disposable pen-shaped capillary gel electrophoresis cartridge for fluorescence detection of bio-molecules

    Science.gov (United States)

    Amirkhanian, Varoujan; Tsai, Shou-Kuan

    2014-03-01

    We introduce a novel and cost-effective capillary gel electrophoresis (CGE) system utilizing disposable pen-shaped gelcartridges for highly efficient, high speed, high throughput fluorescence detection of bio-molecules. The CGE system has been integrated with dual excitation and emission optical-fibers with micro-ball end design for fluorescence detection of bio-molecules separated and detected in a disposable pen-shaped capillary gel electrophoresis cartridge. The high-performance capillary gel electrophoresis (CGE) analyzer has been optimized for glycoprotein analysis type applications. Using commercially available labeling agent such as ANTS (8-aminonapthalene-1,3,6- trisulfonate) as an indicator, the capillary gel electrophoresis-based glycan analyzer provides high detection sensitivity and high resolving power in 2-5 minutes of separations. The system can hold total of 96 samples, which can be automatically analyzed within 4-5 hours. This affordable fiber optic based fluorescence detection system provides fast run times (4 minutes vs. 20 minutes with other CE systems), provides improved peak resolution, good linear dynamic range and reproducible migration times, that can be used in laboratories for high speed glycan (N-glycan) profiling applications. The CGE-based glycan analyzer will significantly increase the pace at which glycoprotein research is performed in the labs, saving hours of preparation time and assuring accurate, consistent and economical results.

  10. Urine Metabolite Profiling of Human Colorectal Cancer by Capillary Electrophoresis Mass Spectrometry Based on MRB

    Directory of Open Access Journals (Sweden)

    Jin-Lian Chen

    2012-01-01

    (P<0.05. Conclusion. The technique of capillary electrophoresis mass spectrometry based on MRB could reveal the significant metabolic alterations during progression of colorectal cancer, and the method is feasible and may be useful for the early diagnosis of colorectal cancer.

  11. Assessment of genetic diversity in Saccharum using SSR markers and capillary electrophoresis

    Science.gov (United States)

    This study was conducted to assess the genetic diversity amongst 12 Saccharum clones from 3 species using SSR markers and CE (capillary electrophoresis). Genomic DNA of 12 sugarcane cultivars was amplified with 19 SSR primer pairs. A total of 229 bands generated with a size range between 100 and 26...

  12. Gold Nanoparticles Enhanced Microchip Capillary Electrophoresis for Detection of Serum Lipoprotein

    Institute of Scientific and Technical Information of China (English)

    WANG Hua; WANG DaXin; CAO Li; CHEN Xia

    2009-01-01

    @@ We describe here the use of gold nanoparticles (AuNPs) in conjunction with chip-based capillary electrophoresis (CE) to improve the selectivity between lipoprotein fractions and increase the efficiency of the separation.AuNPs were added into the running buffer to manipulate solution and control the electroosmotic flow (EOF).

  13. Characterization of the Interaction between Bovine Serum Albumin and Lomefloxacin by Capillary Zone Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    Ming GUO; Qing Sen YU; Jian Wei YAN; Fei TAN; Guo Zheng MA

    2004-01-01

    Three capillary zone electrophoresis (CZE) methods of the frontal analysis (FA), vacancy peak (VP) and simplified Hummel-Dreyer (SHD) were applied to investigate interaction between bovine serum albumin (BSA) and lomefloxacin, the experimental condition was established after a large number of tests. Based on the site-binding model, the binding parameters were measured according to the site model by Scatchard.

  14. Improving the reproducibility in capillary electrophoresis by incorporating current drift in mobility and peak area calculations

    DEFF Research Database (Denmark)

    Petersen, Nickolaj J.; Hansen, Steen H

    2012-01-01

    The traditional way of calculating mobility and peak areas in capillary electrophoresis does not take into account the changes in the buffer viscosity at different thermostatic control and that the analytes may accelerate during the individual runs due to Joule heating effects. We present a method...

  15. Capillary electrophoresis of FITC labeled amino acids with laser-induced fluorescence detection

    Institute of Scientific and Technical Information of China (English)

    党福全; 陈义

    1999-01-01

    FITC labeled amino acids have been separated using a home-huilt capillary electrophoresis with a laserinduced fluorescence detection (CE-LIF) system. Seventeen peaks can now be generated from the twenty common amino acids. The key conditions lie in the optimization of pH, buffer electrolytes and buffer additives.

  16. Constant pressure-assisted head-column field-amplified sample injection in combination with in-capillary derivatization for enhancing the sensitivity of capillary electrophoresis.

    Science.gov (United States)

    Yan, Na; Zhou, Lei; Zhu, Zaifang; Zhang, Huige; Zhou, Ximin; Chen, Xingguo

    2009-05-15

    In this work, a novel method combining constant pressure-assisted head-column field-amplified sample injection (PA-HC-FASI) with in-capillary derivatization was developed for enhancing the sensitivity of capillary electrophoresis. PA-HC-FASI uses an appropriate positive pressure to counterbalance the electroosmotic flow in the capillary column during electrokinetic injection, while taking advantage of the field amplification in the sample matrix and the water of the "head column". Accordingly, the analytes were stacked at the stationary boundary between water and background electrolyte. After 600s PA-HC-FASI, 4-fluoro-7-nitro-2,1,3-benzoxadiazole as derivatization reagent was injected, followed by an electrokinetic step (5kV, 45s) to enhance the mixing efficiency of analytes and reagent plugs. Standing a specified time of 10min for derivatization reaction under 35 degrees C, then the capillary temperature was cooled to 25 degrees C and the derivatives were immediately separated and determined under 25 degrees C. By investigating the variables of the presented approach in detail, on-line preconcentration, derivatization and separation could be automatically operated in one run and required no modification of current CE commercial instrument. Moreover, the sensitivity enhancement factor of 520 and 800 together with the detection limits of 16.32 and 6.34pg/mL was achieved for model compounds: glufosinate and aminomethylphosphonic acid, demonstrating the high detection sensitivity of the presented method. PMID:19342058

  17. A low-makeup beveled tip capillary electrophoresis /electrospray ionization mass spectrometry interface for micellar electrokinetic chromatography and nonvolatile buffer capillary electrophoresis.

    Science.gov (United States)

    Tseng, Mei-Chun; Chen, Yet-Ran; Her, Guor-Rong

    2004-11-01

    A robust interface has been developed for interfacing micellar electrokinetic chromatography (MEKC) and nonvolatile buffer capillary electrophoresis (CE) to electrospray ionization mass spectrometry (ESI-MS). The interface consists of two parallel capillaries for separation (50 microm i.d. x 155 microm o.d.) and makeup (50 microm i.d. x 155 microm o.d.) housed within a larger capillary (530 microm i.d. x 690 microm o.d.). The capillaries terminate in a single tapered tip having a beveled edge. The use of a tapered beveled edge results in a greater tip orifice diameter (75 microm) than in a previous design from our laboratory (25 microm) that used a flat tip. While maintaining a similar optimum flow rate and consequently similar sample dilution, a 75-microm beveled emitter is more rugged than a 25-microm flat tip. Furthermore, the incorporation of a sheath liquid capillary allows the compositions of the final spray solution to be controlled. The application of this novel CE/ESI-MS interface was demonstrated for MEKC using mixtures of triazines (positive ion mode) and phenols (negative ion mode). The ability to perform CE/ESI-MS using a nonvolatile buffer was demonstrated by the analysis of gangliosides with a buffer consisting of 40 mM borate and 20 mM alpha-cyclodextrin.

  18. Technology to accelerate pangenomic scanning for unknown point mutations in exonic sequences: cycling temperature capillary electrophoresis (CTCE

    Directory of Open Access Journals (Sweden)

    Bjørheim Jens

    2007-08-01

    Full Text Available Abstract Background Rapid means to discover and enumerate unknown mutations in the exons of human genes on a pangenomic scale are needed to discover the genes carrying inherited risk for common diseases or the genes in which somatic mutations are required for clonal diseases such as atherosclerosis and cancers. The method of constant denaturing capillary electrophoresis (CDCE permitted sensitive detection and enumeration of unknown point mutations but labor-intensive optimization procedures for each exonic sequence made it impractical for application at a pangenomic scale. Results A variant denaturing capillary electrophoresis protocol, cycling temperature capillary electrophoresis (CTCE, has eliminated the need for the laboratory optimization of separation conditions for each target sequence. Here are reported the separation of wild type mutant homoduplexes from wild type/mutant heteroduplexes for 27 randomly chosen target sequences without any laboratory optimization steps. Calculation of the equilibrium melting map of each target sequence attached to a high melting domain (clamp was sufficient to design the analyte sequence and predict the expected degree of resolution. Conclusion CTCE provides practical means for economical pangenomic detection and enumeration of point mutations in large-scale human case/control cohort studies. We estimate that the combined reagent, instrumentation and labor costs for scanning the ~250,000 exons and splice sites of the ~25,000 human protein-coding genes using automated CTCE instruments in 100 case cohorts of 10,000 individuals each are now less than U.S. $500 million, less than U.S. $500 per person.

  19. Fast separation and analysis of reduced monoclonal antibodies with capillary zone electrophoresis coupled to mass spectrometry.

    Science.gov (United States)

    Zhao, Yimeng; Sun, Liangliang; Knierman, Michael D; Dovichi, Norman J

    2016-02-01

    Capillary zone electrophoresis-electrospray ionization-mass spectrometry (CZE-ESI-MS) was used for analysis of reduced antibodies. We first developed a simple protocol to condition commercial linear-polyacrylamide coated capillaries for use in top-down proteomics. We then suspended reduced antibodies in a solution of 35% acetic acid, 50% acetonitrile in water. Heavy and light chains were baseline resolved within 10 min and with 3-30 µg/mL detection limits using a 0.1% aqueous formic acid background electrolyte. Quintuplicate runs of a two-antibody mixture produced relative standard deviations of ∼1% in migration time and 10% in peak amplitudes. Resolution was further improved for the two-antibody mixture by using 5% acetic acid as the background electrolyte, highlighting the potential of capillary electrophoresis-mass spectrometry for analysis of antibody mixtures. PMID:26653481

  20. Affinity chromatography and capillary electrophoresis for analysis of the yeast ribosomal proteins

    Directory of Open Access Journals (Sweden)

    Miriam S. Goyder

    2012-04-01

    Full Text Available We present a top down separation platform for yeast ribosomalproteins using affinity chromatography and capillary electrophoresiswhich is designed to allow deposition of proteins ontoa substrate. FLAG tagged ribosomes were affinity purified, andrRNA acid precipitation was performed on the ribosomes followedby capillary electrophoresis to separate the ribosomalproteins. Over 26 peaks were detected with excellent reproducibility(<0.5% RSD migration time. This is the first reportedseparation of eukaryotic ribosomal proteins using capillaryelectrophoresis. The two stages in this workflow, affinity chromatographyand capillary electrophoresis, share the advantagesthat they are fast, flexible and have small sample requirementsin comparison to more commonly used techniques. This methodis a remarkably quick route from cell to separation that hasthe potential to be coupled to high throughput readout platformsfor studies of the ribosomal proteome. [BMB reports2012; 45(4: 233-238

  1. Capillary electrophoresis: Biotechnology for separation of DNA and chromosomes

    Science.gov (United States)

    Williams, George O., Jr.

    1994-01-01

    Electrophoresis has been used for the separation of particles, ions, and molecules for a number of years. The technology for separation and detection of the results has many applications in the life sciences. One of the major goals of the scientific community is to separate DNA molecules and intact chromosomes based upon their different lengths or number of base pairs. This may be achieved by using some of the commercially available and widely used methods, but these processes require a considerable amount of time. The challenge is to achieve separation of intact chromosomes in a short time, preferably in a matter of minutes.

  2. A Novel Polybrene/Chondroitin Sulfate C Double Coated Capillary and Its Application in Capillary Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    DU,Ying-Xiang(杜迎翔); HONDA,Susumu; TAGA,Atsushi; LIU,Wen-Ying(刘文英); SUZUKI,Shigeo

    2002-01-01

    A new capillary coated by double polymer, polybrene/chondroitin sulfate C (P/CC), was developed using a simple procedure. The P/CC double coated capillary showed long lifetime,strong chemical stability and good reproducibility. It endured during more than 100 replicated analyses and was also tolerant to HCl (1 mol/L), NaOH (0.01 mol/L), CH3OH and CH3CN. The P/CC double coated capillary can be applied to basic drug analyses. The adsorption of basic drugs to the capillary wall was suppressed and the peak tailing greatly decreased. The use of the P/CC double coated capillary allowed excelent separation of the enantiomers of some basic drugs by using chondroitin sulfate C as the chiral selector, ami the peak symmetry of basic drugs was further improved under these conditions.

  3. Direct identification of all oncogenic mutants in KRAS exon 1 by cycling temperature capillary electrophoresis.

    Science.gov (United States)

    Bjørheim, Jens; Gaudernack, Gustav; Giercksky, Karl-Erik; Ekstrøm, Per O

    2003-01-01

    Over the past few decades, advances in genetics and molecular biology have revolutionized our understanding of cancer initiation and progression. Molecular progression models outlining genetic events have been developed for many solid tumors, including colon cancer. Previous reports in the literature have shown a relationship between different KRAS mutations and prognosis and response to medical treatment in colon cancer patients. Furthermore, the presence of a mutated KRAS has been correlated with different clinicopathological variables including age and gender of patients and tumor location. To our knowledge, few institutions screen for KRAS mutations on regular basis in colon cancer patients despite such evidence that knowledge of KRAS exon 1 status is informative. Here, we report on a mutation analysis method adapted to a 96-capillary electrophoresis instrument that allows identification of all 12 oncogenic mutations in KRAS exon 1 under denaturing conditions. To determine the optimal parameters, a series of DNA constructs generated by site-directed mutagenesis was analyzed and the migration times of all mutant peaks were measured. A classification tree was then made based on the differences in migration time between the mutants and an internal standard. A randomized series of 500 samples constructed with mutagenesis as well as 60 blind samples from sporadic colon carcinomas was analyzed to test the method. No wild-type samples were scored as mutants and all mutants were correctly identified. Post polymerase chain reaction (PCR) analysis time of 96 samples was performed within 40 min. PMID:12652573

  4. Application of capillary electrophoresis to the development and evaluation of aptamer affinity probes

    Science.gov (United States)

    Sooter, Letha J.; McMasters, Sun; Stratis-Cullum, Dimitra N.

    2007-09-01

    Nucleic acid aptamers can exhibit high binding affinities for a wide variety of targets and have received much attention as molecular recognition elements for enhanced biosensor performance. These aptamers recognize target molecules through a combination of conformational dependent non-covalent interactions in aqueous media which can be investigated using capillary electrophoresis-based methods. In this paper we report on the results of our studies of the relative binding affinity of Campylobacter jejuni aptamers using a capillary electrophoretic immunoassay. Our results show preferential binding to C. jejuni over other common food pathogen bacteria. Capillary electrophoresis can also be used to develop new aptamer recognition elements using an in vitro selection process known as systematic evolution of ligand by exponential enrichment (SELEX). Recently, this process has been adapted to use capillary electrophoresis in an attempt to shorten the overall selection process. This smart selection of nucleic acid aptamers from a large diversity of a combinatorial DNA library is under optimization for the development of aptamers which bind to Army-relevant targets. This paper will include a discussion of the establishment of CE-SELEX methods for the future development of smart aptamer probes.

  5. Recent advances in the analysis of therapeutic proteins by capillary and microchip electrophoresis.

    Science.gov (United States)

    Creamer, Jessica S; Oborny, Nathan J; Lunte, Susan M

    2014-07-01

    The development of therapeutic proteins and peptides is an expensive and time-intensive process. Biologics, which have become a multi-billion dollar industry, are chemically complex products that require constant observation during each stage of development and production. Post-translational modifications along with chemical and physical degradation from oxidation, deamidation, and aggregation, lead to high levels of heterogeneity that affect drug quality and efficacy. The various separation modes of capillary electrophoresis (CE) are commonly utilized to perform quality control and assess protein heterogeneity. This review attempts to highlight the most recent developments and applications of CE separation techniques for the characterization of protein and peptide therapeutics by focusing on papers accepted for publication in the in the two-year period between January 2012 and December 2013. The separation principles and technological advances of CE, capillary gel electrophoresis, capillary isoelectric focusing, capillary electrochromatography and CE-mass spectrometry are discussed, along with exciting new applications of these techniques to relevant pharmaceutical issues. Also included is a small selection of papers on microchip electrophoresis to show the direction this field is moving with regards to the development of inexpensive and portable analysis systems for on-site, high-throughput analysis.

  6. Advances in Automation and Throughput of the Mars Organic Analyzer Microchip Capillary Electrophoresis System

    Science.gov (United States)

    Haldeman, B. J.; Skelley, A. M.; Scherer, J. R.; Jayarajah, C.; Mathies, R. A.

    2005-12-01

    We have previously demonstrated the design, construction and testing of a portable microchip capillary electrophoresis (CE) instrument called the Mars Organic Analyzer (MOA) for analysis of amino acids and amine containing organic molecules (1). This instrument is designed to accept organic compounds isolated from samples by sublimation or by subcritical water extraction, to label the amine groups with fluorescamine, and to perform high resolution electrophoretic analysis. The CE instrument has shown remarkable robustness during successful field tests last year in the Panoche Valley, CA (1) and more recently in the Atacama Desert, Chile (2). For successful operation on Mars, however, it is necessary to operate autonomously and to analyze large numbers of samples, blanks, and standards. Toward this end we present here two advances in the MOA system that test key aspects of an eventual flight prototype. First, we have developed an automated microfluidic system and method for the autonomous loading, running and cleaning of the CE chip on the single channel MOA instrument. The integration of microfabricated PDMS valves and pumps with all-glass separation channels in a multilayer design enabled creation of structures for complex fluidic routing. Twenty sequential analyses of an amino acid standard were performed with an automated cleaning procedure between runs. In addition, dilutions were performed on-chip, and blanks were run to demonstrate the elimination of carry-over from run to run. These results demonstrate an important advance of the technology readiness level of the MOA. Second, we have designed, constructed and successfully tested a lab version of the multichannel instrument we initially proposed for the MSL opportunity. The portable Multi-Channel Mars Organic Analyzer (McMOA, 25 by 30 by 15 cm), was designed to sequentially interrogate eight radially oriented CE separation channels on a single wafer. Since each channel can be used to analyze 20 or more

  7. Quantitation of Leishmania lipophosphoglycan repeat units by capillary electrophoresis.

    Science.gov (United States)

    Barron, Tamara L; Turco, Salvatore J

    2006-04-01

    The glycosylphosphatidylinositol (GPI)-anchored lipophosphoglycan (LPG) of Leishmania is the dominant cell surface glycoconjugate of these pathogenic parasites. LPG is structurally characterized by a series of phosphoglycan repeat units. Determining the number of repeat units per LPG molecule has proven difficult using current technologies, such as mass spectrometry. As an alternative method to quantitate the number of repeat units in LPG, a procedure based on capillary electrophoretic analysis of the proportion of mannose to 2,5-anhydromannose (derived from the nonacetylated glucosamine of the GPI anchor of LPG) was developed. The CE-based technique is sensitive and relatively rapid compared to GC-MS-based protocols. Its application was demonstrated in quantitating the number of LPG repeat units from several species of Leishmania as well as from two life-cycle stages of these organisms. PMID:16310310

  8. Gel versus capillary electrophoresis genotyping for categorizing treatment outcomes in two anti-malarial trials in Uganda

    Directory of Open Access Journals (Sweden)

    Hubbard Alan E

    2010-01-01

    Full Text Available Abstract Background Molecular genotyping is performed in anti-malarial trials to determine whether recurrent parasitaemia after therapy represents a recrudescence (treatment failure or new infection. The use of capillary instead of agarose gel electrophoresis for genotyping offers technical advantages, but it is unclear whether capillary electrophoresis will result in improved classification of anti-malarial treatment outcomes. Methods Samples were genotyped using both gel and capillary electrophoresis from randomized trials of artemether-lumefantrine (AL vs. dihydroartemisinin-piperaquine (DP performed in two areas of Uganda: Kanungu, where transmission is moderate, and Apac, where transmission is very high. Both gel and capillary methods evaluated polymorphic regions of the merozoite surface protein 1 and 2 and glutamine rich protein genes. Results Capillary electrophoresis detected more alleles and provided higher discriminatory power than agarose gel electrophoresis at both study sites. There was only moderate agreement between classification of outcomes with the two methods in Kanungu (kappa = 0.66 and poor agreement in Apac (kappa = 0.24. Overall efficacy results were similar when using gel vs. capillary methods in Kanungu (42-day risk of treatment failure for AL: 6.9% vs. 5.5%, p = 0.4; DP 2.4% vs. 2.9%, p = 0.5. However, the measured risk of recrudescence was significantly higher when using gel vs. capillary electrophoresis in Apac (risk of treatment failure for AL: 17.0% vs. 10.7%, p = 0.02; DP: 8.5% vs. 3.4%, p = 0.03. Risk differences between AL and DP were not significantly different whether gel or capillary methods were used. Conclusions Genotyping with gel electrophoresis overestimates the risk of recrudescence in anti-malarial trials performed in areas of high transmission intensity. Capillary electrophoresis provides more accurate outcomes for such trials and should be performed when possible. In areas of moderate transmission

  9. Determination of aggregation thresholds of UV absorbing anionic surfactants by frontal analysis continuous capillary electrophoresis.

    Science.gov (United States)

    Le Saux, Thomas; Varenne, Anne; Gareil, Pierre

    2004-06-01

    Aggregation of anionic surfactants was investigated by frontal analysis continuous capillary electrophoresis (FACCE), a method involving the continuous electrokinetic introduction of the surfactant sample into the separation capillary. This process results in a partial separation of the monomeric and aggregated forms without perturbing the monomer-aggregate equilibrium. The critical micelle concentration (CMC) can then be easily derived from the height of the firstly detected migration front, corresponding to the monomeric form. This approach is exemplified with octyl and dodecylbenzenesulfonates and compared with conductimetry and surface tension measurements. FACCE turns out to be an effective method for the determination of CMC and intermediate aggregation phenomena with very small sample and short time requirements.

  10. Determination of phenolic acids in olive oil by capillary electrophoresis.

    Science.gov (United States)

    Buiarelli, Francesca; Di Berardino, Sonia; Coccioli, Franco; Jasionowska, Renata; Russo, Mario Vincenzo

    2004-01-01

    A CZE method for the separation and quantitation of phenolic acids (cinnamic, syringic, p-coumaric, vanillic, caffeic, 3,4-dihydroxyphenylacetic, protocatechuic), extracted from extra virgin olive oil, was developed. The sample preparation involved the LLE and SPE extraction methods. CE separation was performed in a fused silica capillary of I.D.= 50microm using as a BGE 40 mM borate buffer at pH=9.2. The separation voltage was 18kV with corresponding current of 27-28 microA. Detection was accomplished with UV-detector at lambda=200nm. The proposed method was fully validated. A good repeatability of migration time (RSD% ranged from 0.81 to 1.63) and of corrected peak area (RSD% from 2.89 to 5.77) was obtained. The linearity of detector response in the range from 5 to 50 ppm was checked, obtaining the correlation coefficient R2 values in the range: 0.9919-0.9997. Some phenolic acids in real oil samples were detected and quantified with the proposed method. PMID:15506620

  11. Nonaqueous capillary electrophoresis of imatinib mesylate and related substances.

    Science.gov (United States)

    Ye, Lei; Huang, Yifei; Li, Jian; Xiang, Guangya; Xu, Li

    2012-08-01

    In the present study, nonaqueous capillary electrophoretic separation of imatinib mesylate (IM) and related substances, N-(5-amino-2-methylphenyl)-4-(3-pyridyl)-2-pyrimidinamine (PYA), N-(4-methyl-3-(4-(pyridin-3-yl)pyrimidin-2-ylamino)phenyl)-4-((piperazin-1-yl)methyl) benzamide (NDI) and 4-chloromethyl-N-(4-methyl-3-((4-(pyridin-3-yl) pyrimidin-2-yl) amino) phenyl) benzamide (CPB) was developed. The influential factors affecting separation, including type and concentration of the electrolyte, applied voltage, and buffer modifier were investigated. Baseline separation of the studied analytes was obtained using a buffer of 50 mM Tris and 50 mM methanesulfonic acid in methanol at a apparent pH (pH*) of 1.65. To enhance the sensitivity, large-volume sample stacking was employed for online concentration. The strongest analytical signal with a suitable separation was achieved when the injection time was 100 s. The linearity ranges of PYA and NDI were 0.100-2.50 μg mL(-1), and that of CPB was 0.125-2.50 μg mL(-1), with good coefficients (r(2) > 0.9948). The relative standard deviations of intra- and interday were satisfactory. Under the optimized conditions, seven batches of the synthesized samples were analyzed and CPB was detected in two batches. Owing to its simplicity, effectiveness, and low price, the developed method is promising for quality control of IM.

  12. Analysis of flavonoids by capillary zone electrophoresis with electrokinetic supercharging.

    Science.gov (United States)

    Zhong, Hao; Yao, Qingqiang; Breadmore, Michael C; Li, Yumei; Lu, Yuanqi

    2011-11-01

    On-line concentration via Electrokinetic Supercharging (EKS) was used to enhance the sensitivity of the capillary electrophoretic separation of the four flavonoids naringenin, hesperetin, naringin and hesperidin. Separation conditions, including the background electrolyte pH and concentration, the length and choice of terminator and the electrokinetic injection time were optimized. The optimum conditions were: a background electrolyte of 30 mM sodium tetraborate (pH 9.5) containing 5% (v/v) of methanol, electrokinetic injection of the sample (130 s, -10 kV) followed by hydrodynamic injecting of 100 mM 2-(cyclohexylamino)ethanesulfonic acid (CHES) (17 s, 0.5 psi) as terminator, and separation with -20 kV. Under these conditions the four flavonoids could be separated with a sample-to-sample time of 15 min and detection limits from 2.0 to 6.8 ng mL(-1). When compared to a conventional hydrodynamic injection the sensitivity was enhanced between 824 and 1515 times which is 7.6-16 times higher than other CE methods for the on-line concentration of flavonoids. The applicability of the developed method was demonstrated by the detection of the four flavonoids in an aqueous extract of Clematis hexapetala pall. PMID:21949941

  13. Ultrafast Capillary Electrophoresis Isolation of DNA Aptamer for the PCR Amplification-Based Small Analyte Sensing

    Directory of Open Access Journals (Sweden)

    Emmanuelle eFiore

    2015-08-01

    Full Text Available Here, we report a new homogeneous DNA amplification-based aptamer assay for small analyte sensing. The aptamer of adenosine chosen as the model analyte was split into two fragments able to assemble in the presence of target. Primers were introduced at extremities of one fragment in order to generate the amplifiable DNA component. The amount of amplifiable fragment was quantifiable by Real-Time Polymerase Chain Reaction (RT-PCR amplification and directly reliable on adenosine concentration. This approach combines the very high separation efficiency and the homogeneous format (without immobilization of capillary electrophoresis and the sensitivity of real time PCR amplification. An ultrafast isolation of target-bound split aptamer (60 s was developed by designing a capillary electrophoresis input/ouput scheme. Such method was successfully applied to the determination of adenosine with a LOD of 1 µM.

  14. Ultrafast Capillary Electrophoresis Isolation of DNA Aptamer for the PCR Amplification-Based Small Analyte Sensing

    Science.gov (United States)

    Fiore, Emmanuelle; Dausse, Eric; Dubouchaud, Hervé; Peyrin, Eric; Ravelet, Corinne

    2015-08-01

    Here, we report a new homogeneous DNA amplification-based aptamer assay for small analyte sensing. The aptamer of adenosine chosen as the model analyte was split into two fragments able to assemble in the presence of target. Primers were introduced at extremities of one fragment in order to generate the amplifiable DNA component. The amount of amplifiable fragment was quantifiable by Real-Time Polymerase Chain Reaction (RT-PCR) amplification and directly reliable on adenosine concentration. This approach combines the very high separation efficiency and the homogeneous format (without immobilization) of capillary electrophoresis and the sensitivity of real time PCR amplification. An ultrafast isolation of target-bound split aptamer (60 s) was developed by designing a capillary electrophoresis input/ouput scheme. Such method was successfully applied to the determination of adenosine with a LOD of 1 µM.

  15. A New Immunoassay Method by Capillary Electrophoresis with Enhanced Chemiluminescence Detection

    Institute of Scientific and Technical Information of China (English)

    Jiao Ning WANG; Ji Cun REN

    2005-01-01

    This paper described a new immunoassay method by capillary electrophoresis with enhanced chemiluminescence (CL) detection system based on luminol-hydrogen peroxide reaction catalyzed by horseradish peroxides (HRP). Using para-iodophenol as a CL enhancer, the detection limit of about 1×10-12 mol/L for HRP was achieved, which corresponded to 1.32×10-5U/mL. In optimal conditions, the free HRP-labeled CA125 antibody (Ab*) and the bound enzyme-labeled complex (Ab*-Ag) were well separated by capillary electrophoresis within 4 min.The assay was successfully used to determine the contents of CA125 in human sera, which were associated with ovarian cancer, and the recoveries of the standard addition experiments were 96 to109 %.

  16. Steroid determination in fish plasma using capillary electrophoresis

    Science.gov (United States)

    Bykova, L.; Archer-Hartmann, S. A.; Holland, L.A.; Iwanowicz, L.R.; Blazer, V.S.

    2010-01-01

    A capillary separation method that incorporates pH-mediated stacking is employed for the simultaneous determination of circulating steroid hormones in plasma from Perca flavescens (yellow perch) collected from natural aquatic environments. The method can be applied to separate eight steroid standards: progesterone, 17α,20β-dihydroxypregn-4-en-3-one, 17α-hydroxyprogesterone, testosterone, estrone, 11-ketotestosterone, ethynyl estradiol, and 17β-estradiol. Based on screening of plasma, the performance of the analytical method was determined for 17α,20β-dihydroxypregn-4-en-3-one, testosterone, 11-ketotestosterone, and 17β-estradiol. The within-day reproducibility in migration time for these four steroids in aqueous samples was ≤2%. Steroid quantification was accomplished using a calibration curve obtained with external standards. Plasma samples from fish collected from the Choptank and Severn Rivers, Maryland, USA, stored for up to one year were extracted with ethyl acetate and then further processed with anion exchange and hydrophobic solid phase extraction cartridges. The recovery of testosterone and 17β-estradiol from yellow perch plasma was 84 and 85%, respectively. Endogenous levels of testosterone ranged from 0.9 to 44 ng/ml, and when detected 17α,20β-dihydroxypregn-4-en-3-one ranged from 5 to 34 ng/ml. The reported values for testosterone correlated well with the immunoassay technique. Endogenous concentrations of 17β-estradiol were ≤1.7 ng/ml. 11-Ketotestosterone was not quantified because of a suspected interferant. Higher levels of 17α,20β-dihydroxypregn-4-en-3-one were found in male and female fish in which 17β-estradiol was not detected. Monitoring multiple steroids can provide insight into hormonal fluctuations in fish.

  17. Capillary electrophoresis as a tool for screening aptamer with high affinity and high specificity to ricin

    Institute of Scientific and Technical Information of China (English)

    TANG Jijun; XIE Jianwei; SHAO Ningsheng; GUO Lei; YAN Yan

    2007-01-01

    Aptamers which specifically recognize target sare selected from random oligonucleotide library using systematic evolution of ligands by exponential enrichment(SELEX). In this paper, capillary electrophoresis (CE) as a separation approach has been introduced to SELEX pro-cedure. The high efficiency of CE gives rise to greatly shorten the selection procedure. The results from enzyme-linked assay and dot blot experiment show that an enrichment pool has been obtained after four rounds selection, which can specifically recognize riein.

  18. An accessible micro-capillary electrophoresis device using surface-tension-driven flow

    OpenAIRE

    Mohanty, Swomitra K; Warrick, Jay; Gorski, Jack; Beebe, David J.

    2009-01-01

    We present a rapidly fabricated micro-capillary electrophoresis chip that utilizes surface-tension-driven flow for sample injection and extraction of DNA. Surface-tension-driven flow (i.e. passive pumping) injects a fixed volume of sample that can be predicted mathematically. Passive pumping eliminates the need for tubing, valves, syringe pumps, and other equipment typically needed for interfacing with microelectrophoresis chips. This method requires a standard micropipette to load samples be...

  19. Determination of chlorophenols in environmental samples using electromembrane extraction and capillary electrophoresis

    OpenAIRE

    Šlampová, Andrea

    2013-01-01

    Combination of electromembrane extraction (EME) with capillary electrophoresis (CE) was used for determination of trace level chlorophenols (CPs) in environmental water samples. The analytes were transported across supported liquid membrane (SLM), composed of 1-ethyl-2-nitrobenzene (ENB), by the application of electrical field. A driving force of 150 V was applied to extract the analytes from neutral sample (donor solution) into strongly alkaline acceptor solutions. The acceptor soluti...

  20. Determination of Dissociation Constants of Complicated Compounds by Capillary Zone Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    YANG, Geng-Liang; WANG, De-Xian; SUN, Su-Fang; LIU, Hai-Xing; MA, Jian-Jun

    2001-01-01

    In this work,the whole theoretical metods forthe determinaion ofpKa1 and pKa2 of complicated complicated compounds are proposed by capillary zone electrophoresis.The pka values areachieved by non-linear regression analysis by takiny into consideration the effect of activity coefficient.This is the first report on determining the dissociation constants of gastrodin,magnolol,honkiol,puercetin,curcumin,diethylstilbestrol,diehylstilbestrol,4acetamidophenol,eugenol and paeonol.

  1. Aggregate of Amphiphilic Block Copolymer as a Pseudo-Stationary Phase in Capillary Electrophoresis

    OpenAIRE

    Nakamura, Tohru; OHKI, Akira; Mishiro, Masaki; Tsuyashima, Osamu; Maeda, Shigeru; ナカムラ, トオル; オオキ, アキラ; ミシロ, マサキ; ツヤシマ, オサム; マエダ, シゲル; 中村, 透; 大木, 章; 艶島, 修; 前田, 滋

    1999-01-01

    The use of an aggregate of amphiphilic block copolymer 1, which consists of poly[(N-acetylimino)ethylene] and poly[(N-pentanoylimino)ethylene], for a pseudo-stationary phase in capillary electrophoresis has been examined. From gel-filtration chromatography, the aggregate from 1 (1-AG) was found to incorporate phenol. When the running solution contains 1-AG and sodium dodecyl sulfate (SDS), the electrophoretic mobility becomes nearly zero. Thus, it is found that when 1-AG and SDS are added to ...

  2. Further development of capillary electrophoresis for the quantitative determination of small inorganic anions

    OpenAIRE

    King, Marion

    2003-01-01

    Factors influencing the separation and indirect UV absorbance detection of common inorganic anions using capillary zone electrophoresis (CZE) have been investigated. Initially a number of different aspects of indirect background electrolyte (BGE) systems were studied, with the resultant observations indicating the requirements of an 'ideal' BGE system for the separation and detection of common inorganic anions in water samples. In addition to the above the correct use of buffers within BGE us...

  3. Determination of Cordycepin in Cordyceps kyushuensis by Capillary Electrophoresis and its Antitumour Activity

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    A simple, rapid and low-cost method of determination for cordycepin in Cordyceps kyushuensis by capillary zone electrophoresis (CZE) was developed. Based on the finding that there is a high concentration of cordycepin in both natural and cultured Cordyceps kyushuensis, the in vitro antitumor activity of cordycepin and the water extracts of Cordyceps kyushuensis has been investigated. This is the first report about the antitumor effect of Cordyceps kyushuensis.

  4. Development of a capillary electrophoresis method for the simultaneous determination of cephalosporins

    OpenAIRE

    Hancu Gabriel; Kelemen Hajnal; Rusu Aura; Gyéresi Árpád

    2013-01-01

    A rapid and simple capillary electrophoresis method has been developed for the simultaneous determination of six extensively used cephalosporin antibiotics (cefaclor, cefadroxil, cefalexin, cefuroxim, ceftazidim, ceftriaxon). The determination of cephalosporins was performed at a pH 6.8, using a 25 mM phospate - 25 mM borate mixed buffer, + 25 kV voltage at a temperature of 25 °C. We achieved a baseline separation in approximately 10 minutes. The separation resolution was increased by a...

  5. DNA Separation by Capillary Electrophoresis with Ultraviolet Detection using Mixed Synthetic Polymers

    Institute of Scientific and Technical Information of China (English)

    Qian WANG; Xu XU

    2003-01-01

    The mixtures of two polymers, poly (N,N-dimethylacrylamide) (PDMA) and polyvinylpyrrolidone (PVP) were synthesized and used as the separation medium for double-stranded and single-stranded DNA fragments by capillary electrophoresis with UV detector. On optimal conditions, 2%w/v PDMA ( 2%w/v PVP can be used to separate the doublet 123/124bp in pBR322/Hae III Markers.

  6. Cyclodextrine Screening for the Chiral Separation of Amlodipine Enantiomers by Capillary Electrophoresis

    OpenAIRE

    Gabriel Hancu; Monica Budău; Lajos Kristóf Kántor; Anca Cârje

    2015-01-01

    Purpose: Amlodipine is a long acting, dihydropyridine type calcium channel blocker frequently used in the treatment of hypertension and coronary insufficiency. The calcium channel blocking activity resides primarily in the S-amlodipine enantiomer, while R-amlodipine is a potent inhibitor of smooth muscle cell migration. Methods: In this study capillary electrophoresis was applied for the enantiomeric separation of amlodipine using different native and derivatized; neutral and charged cyclo...

  7. Chiral Separation of Ibuprofen and Terbutaline by Nonaqueous Capillary Electrophoresis with Conductance Detection

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    In the nonaqueous N,N-dimethylformamide medium, the chiral drugs ibuprofen and terbutaline were successfully separated with sulfonyl-β-cyclodextrin(s-β-CD) as the chiral selector by capillary electrophoresis with conductance detection. The comparison of the effects of three CDS(β-CD, diethylic-β-CD, sulfonyl-β-CD) on the chiral separation was made and the resolution mechanism was proposed.

  8. A covalent modified hydrophilic capillary for enhanced capillary electrophoresis of biopolymers

    Institute of Scientific and Technical Information of China (English)

    Lian Guo Shan; Xue Yu; Yin Mao Wei; Xiao Hui Zheng; Jian Bin Zheng

    2009-01-01

    δ-Gluconolactone was covalently coupled to aminopropyl derivatized capillary,which created hydrophilic brushes on the inner wall of the capillary.The coated capillary was shown to generate a stable electroosmotic flow(EOF)in the investigated pH range of 2.0-9.0 and to suppress effectively the adsorption of proteins.And it enabled separation of some biopolymer mixtures including basic proteins,DNA and tryptic digested bovine serum albumin(BSA)within 15 min with efficiencies up to 450,000 plates/m.The intra-and inter-day reproducibility of the coating referring to the retention times of proteins were satisfactory with mean relative standard deviations(R.S.D.)of 0.8 and 1.7%,respectively.

  9. Characterization and Study of Transgenic Cultivars by Capillary and Microchip Electrophoresis

    Directory of Open Access Journals (Sweden)

    Elena Domínguez Vega

    2014-12-01

    Full Text Available Advances in biotechnology have increased the demand for suitable analytical techniques for the analysis of genetically modified organisms. Study of the substantial equivalence, discrimination between transgenic and non-transgenic cultivars, study of the unintended effects caused by a genetic modification or their response to diverse situations or stress conditions (e.g., environmental, climatic, infections are some of the concerns that need to be addressed. Capillary electrophoresis (CE is emerging as an alternative to conventional techniques for the study and characterization of genetically modified organisms. This article reviews the most recent applications of CE for the analysis and characterization of transgenic cultivars in the last five years. Different strategies have been described depending on the level analyzed (DNA, proteins or metabolites. Capillary gel electrophoresis (CGE has shown to be particularly useful for the analysis of DNA fragments amplified by PCR. Metabolites and proteins have been mainly separated using capillary zone electrophoresis (CZE using UV and MS detection. Electrophoretic chips have also proven their ability in the analysis of transgenic cultivars and a section describing the new applications is also included.

  10. Characterization and Study of Transgenic Cultivars by Capillary and Microchip Electrophoresis

    Science.gov (United States)

    Domínguez Vega, Elena; Marina, Maria Luisa

    2014-01-01

    Advances in biotechnology have increased the demand for suitable analytical techniques for the analysis of genetically modified organisms. Study of the substantial equivalence, discrimination between transgenic and non-transgenic cultivars, study of the unintended effects caused by a genetic modification or their response to diverse situations or stress conditions (e.g., environmental, climatic, infections) are some of the concerns that need to be addressed. Capillary electrophoresis (CE) is emerging as an alternative to conventional techniques for the study and characterization of genetically modified organisms. This article reviews the most recent applications of CE for the analysis and characterization of transgenic cultivars in the last five years. Different strategies have been described depending on the level analyzed (DNA, proteins or metabolites). Capillary gel electrophoresis (CGE) has shown to be particularly useful for the analysis of DNA fragments amplified by PCR. Metabolites and proteins have been mainly separated using capillary zone electrophoresis (CZE) using UV and MS detection. Electrophoretic chips have also proven their ability in the analysis of transgenic cultivars and a section describing the new applications is also included. PMID:25535077

  11. A Novel Protocol to Analyze Short- and Long-Chain Fatty Acids Using Nonaqueous Microchip Capillary Electrophoresis

    Science.gov (United States)

    Cable, M. L.; Stockton, A. M.; Mora, Maria F; Willis, P. A.

    2013-01-01

    We propose a new protocol to identify and quantify both short- and long-chain saturated fatty acids in samples of astrobiological interest using non-aqueous microchip capillary electrophoresis (micronNACE) with laser induced fluorescence (LIF).

  12. Determination of Enantiomeric Purity of n-Pyrrolidinyl Phenylpropanol by Capillary Electrophoresis Using b-Cyclodextrin Polymer as Chiral Selector

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Enantiomer of n-pyrrolidinyl phenylpropanol was studied by capillary electrophoresis using b-cyclodextrin polymer as chiral selector. We determined the enantiomeric excess value of n-pyrrolidinyl phenylpropanol with RSD 0.48%.

  13. Restriction Enzyme Pattern Analysis of Mycobacteria DNA by Capillary Electrophoresis with Laser-induced Fluorescence Detection

    Institute of Scientific and Technical Information of China (English)

    Li Yuanqian; Wang Guoqing; Mi Jianping; Zhou Ying; Zeng Hongyan; Zhang Chaowu

    2006-01-01

    A new method for rapidly detecting restriction enzyme patterns of Mycobacterium DNA using capillary electrophoresis with laser-induced fluorescence detection (CE-LIFD)was developed.Polymerase chain reaction was used to amplify a 439-bp fragment of a 65,000-kDa(Mr)heat shock protein gene(hsp65)of Mycobacterium.After digesting amplification products by BstEII and HaeIII,patterns of enzyme cleavage products were detected by both CE-LIFD and agarose gel electrophoresis(AGE),respectively.Experimental parameters of CE were optimized.Restriction enzyme patterns of Mycobacterium DNA were detected in optimum electrophoresis conditions:a coated capillary column with a length of 50 cm and an internal diameter of 100 μm,an electrophoresis buffer of 45 mmol/1 Tris-boric acid-ethylenediaminetetraacetic acid,and a running voltage of 11 kV.The restriction enzyme patterns for eight species of mycobacteria were studied.Relative standard deviations of the relative migration times of DNA segments were<3.6%.Compared with AGE,CE is more outstanding in resolution and detection time,and it can be applied as a more effective means to DNA restriction enzyme pattern analysis.

  14. Development of novel separation techniques for biological samples in capillary electrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Chang, H.T.

    1994-07-27

    This dissertation includes three different topics: general introduction of capillary electrophoresis (CE); gradient in CE and CE in biological separations; and capillary gel electrophoresis (CGE) for DNA separation. Factors such as temperature, viscosity, pH, and the surface of capillary walls affecting the separation performance are demonstrated. A pH gradient between 3.0 and 5.2 is useful to improve the resolution among eight different organic acids. A flow gradient due to the change in the concentration of surfactant, which is able to coat to the capillary wall to change the flow rate and its direction, is also shown as a good way to improve the resolution for organic compounds. A temperature gradient caused by joule heat is shown by voltage programming to enhance the resolution and shorten the separation time for several phenolic compounds. The author also shows that self-regulating dynamic control of electroosmotic flow in CE by simply running separation in different concentrations of surfactant has less matrix effect on the separation performance. One of the most important demonstrations in this dissertation is that the author proposes on-column reaction which gives several advantages including the use of a small amount of sample, low risk of contamination, and time saving and kinetic features. The author uses this idea with laser induced fluorescence (LIF) as a detection mode to detect an on-column digestion of sub-ng of protein. This technique also is applied to single cell analysis in the group.

  15. Feasibility of applying the new improved PCR/LDR/capillary electrophoresis on prenatal diagnosis of fetal beta thalassaemia

    OpenAIRE

    Liao, Xi; Yi, Ping; Zheng, Ying-Ru; Yu, Li-Li; Zhong, Xiao-Lin; Yin-hu HUANG; Han, Lei; Yu-juan ZHU; LI Li

    2014-01-01

    Objective  To investigate the feasibility of applying new improved polymerase chain reaction (PCR)/ligase detection reaction (LDR)/capillary electrophoresis to the prenatal diagnosis of fetal β thalassemia from maternal plasma. Methods  The mixture of different concentrations of normal DNA and trace β-thalassemia mutation heterozygous DNA were used for PCR, and the amplification products obtained thereafter were used for LDR. The ligation product was detected by capillary electrophoresis to d...

  16. On-line wall-free cell for laser-induced fluorescence detection in capillary electrophoresis.

    Science.gov (United States)

    Yu, Chang-Zhu; He, You-Zhao; Xie, Hai-Yang; Gao, Yong; Gan, Wu-Er; Li, Jun

    2009-05-15

    A wall-free detection method based on liquid junction in a capillary gap was proposed for laser-induced fluorescence (LIF) of capillary electrophoresis (CE). The capillary gap of the wall-free cell was fabricated by etching a 10-mm x 50-microm I.D. fused-silica capillary to obtain a polyimide coating sleeve, decoating about 6mm at one end of both 50 microm I.D. separation and liquid junction capillary, inserting the treated capillary ends into the coating sleeve oppositely, fixing the capillaries with a gap distance of 140 microm by epoxy glue and removing the coating sleeve by burning. The theoretical model, experimental results and wall-free cell images indicated that the gap distance and applied voltage were main influence factors on the wall-free detection. Since the wall-free cell increased the absorption light path and avoided the stray light from the capillary wall, it improved the ratio of signal to noise and limit of detection (LOD) of CE-LIF. Three flavin compounds of riboflavin (RF), flavin mononucleotide sodium (FMN) and flavin adenine dinucleotide disodium (FAD) were used to evaluate the wall-free detection method. Compared with on-column cell, the LODs of the wall-free cell were improved 15-, 6- and 9-fold for RF, FMN and FAD, respectively. The linear calibration concentrations of the flavins ranged from 0.005 to 5.0 micromol/L. The column efficiency was in the range from 1.0 x 10(5) to 2.5 x 10(5) plates. The wall-free detection of CE-LIF was applied to the analysis of the flavins in spinach and lettuce leaves. PMID:19329123

  17. Linearity evaluation in absorbance detection: the use of light-emitting diodes for on-capillary detection in capillary electrophoresis.

    Science.gov (United States)

    Macka, M; Andersson, P; Haddad, P R

    1996-12-01

    A model which takes into account both stray light and polychromatic light was used to predict and evaluate linearity in on-capillary detection in capillary electrophoresis (CE). According to the model the stray light is the major factor which determines linearity under typical CE operating conditions. By calculating theoretical absorbance versus concentration plots, the influence of different levels of stray light and polychromatic light on linearity is demonstrated. Experimentally, six light-emitting diodes (LEDs) in the range from 563 to 654 nm were examined as light sources for on-capillary detection in CE. Fitting theoretical curves to measured linearity plots enabled determination of the values of both effective path length and stray light for a particular detection system. The detector linearity for the four LEDs was compared to mercury and tungsten lamps used with interference filters. For potassium permanganate as the test compound, the linear range for a 563 nm LED was two times greater than that for a mercury lamp operated at 546 nm. The relatively poor linearity of the mercury lamp detector is explained by its high level of stray light. The noise of the LED563-based detector was the same as for the mercury lamp, whereas the other LEDs of higher light intensity gave approximately half the noise of the mercury lamp. The lowest noise level of 3 x 10(-5) AU was obtained for the LED at 554 nm (determined at a detector time constant of 0.1 s). PMID:9034772

  18. CSE-MECC two-dimensional capillary electrophoresis analysis of proteins in the mouse tumor cell (AtT-20) homogenate

    OpenAIRE

    Chen, Xingguo; Fazal, Md. Abul; Dovichi, Norman J.

    2007-01-01

    Two-dimensional capillary electrophoresis was used for the separation of proteins and biogenic amines from the mouse AtT-20 cell line. The first-dimension capillary contained a TRIS-CHES-SDS-dextran buffer to perform capillary sieving electrophoresis, which is based on molecular weight of proteins. The second-dimension capillary contained a TRIS-CHES-SDS buffer for micel1ar electrokinetic capillary chromatography. After a 61 seconds preliminary separation, fractions from the first-dimension c...

  19. A Theoretical Analysis of the Influence of Electroosmosis on the Effective Ionic Mobility in Capillary Zone Electrophoresis

    Science.gov (United States)

    Hijnen, Hens

    2009-01-01

    A theoretical description of the influence of electroosmosis on the effective mobility of simple ions in capillary zone electrophoresis is presented. The mathematical equations derived from the space-charge model contain the pK[subscript a] value and the density of the weak acid surface groups as parameters characterizing the capillary. It is…

  20. Protein Cross-Linking Capillary Electrophoresis for Protein-Protein Interaction Analysis.

    Science.gov (United States)

    Ouimet, Claire M; Shao, Hao; Rauch, Jennifer N; Dawod, Mohamed; Nordhues, Bryce; Dickey, Chad A; Gestwicki, Jason E; Kennedy, Robert T

    2016-08-16

    Capillary electrophoresis (CE) has been identified as a useful platform for detecting, quantifying, and screening for modulators of protein-protein interactions (PPIs). In this method, one protein binding partner is labeled with a fluorophore, the protein binding partners are mixed, and then, the complex is separated from free protein to allow direct determination of bound to free ratios. Although it possesses many advantages for PPI studies, the method is limited by the need to have separation conditions that both prevent protein adsorption to capillary and maintain protein interactions during the separation. In this work, we use protein cross-linking capillary electrophoresis (PXCE) to overcome this limitation. In PXCE, the proteins are cross-linked under binding conditions and then separated. This approach eliminates the need to maintain noncovalent interactions during electrophoresis and facilitates method development. We report PXCE methods for an antibody-antigen interaction and heterodimer and homodimer heat shock protein complexes. Complexes are cross-linked by short treatments with formaldehyde after reaching binding equilibrium. Cross-linked complexes are separated by electrophoretic mobility using free solution CE or by size using sieving electrophoresis of SDS complexes. The method gives good quantitative results; e.g., a lysozyme-antibody interaction was found to have Kd = 24 ± 3 nM by PXCE and Kd = 17 ± 2 nM using isothermal calorimetry (ITC). Heat shock protein 70 (Hsp70) in complex with bcl2 associated athanogene 3 (Bag3) was found to have Kd = 25 ± 5 nM by PXCE which agrees with Kd values reported without cross-linking. Hsp70-Bag3 binding site mutants and small molecule inhibitors of Hsp70-Bag3 were characterized by PXCE with good agreement to inhibitory constants and IC50 values obtained by a bead-based flow cytometry protein interaction assay (FCPIA). PXCE allows rapid method development for quantitative analysis of PPIs. PMID:27434096

  1. Application of capillary electrophoresis to anion speciation in soil water extracts: 2. Arsenic

    Energy Technology Data Exchange (ETDEWEB)

    Naidu, R.; Smith, J.; McLaren, R.G.; Stevens, D.P.; Sumner, M.E.; Jackson, P.E.

    2000-02-01

    A method has been developed for the speciation of arsenic (AsO{sub 2}{sup {minus}}, AsO{sub 4}{sup 3{minus}}, and dimethylarsinic [DMA]) in natural soil solutions from contaminated sites in Australia. The separation of these anions was achieved by capillary zone electrophoresis (CZE) using a fused silica capillary with a basic chromate buffer and on-column indirect UV detection at 254 nm. Method parameters, such as electrolyte pH, run voltage, and capillary temperature were studied in order to establish suitable analytical conditions. The ideal separation for As(III) and DMA was achieved with a buffer pH of 8.0, a run voltage of 25 kV, and a capillary temperature of 30 C. Under these conditions, As(V) and orthophosphate ions comigrated. However, the use of a chromate buffer at pH 10, a run voltage of 20 kV, and capillary temperature of 20 C led to complete separation of As(V) and phosphate peaks. Results of these investigations together with recovery test data suggest that separation of the As species from soil solutions can be achieved in less than 5 min with detection limits of 0.50, 0.10, and 0.10 mg L{sup {minus}1} for As(III), As(V), and DMA, respectively.

  2. Impact of capillary conditioning and background electrolyte composition on capillary electrophoresis analysis of prostate specific antigen isoforms.

    Science.gov (United States)

    Farina-Gomez, Noemi; Puerta, Angel; Gonzalez, Monica; Diez-Masa, Jose Carlos; de Frutos, Mercedes

    2016-04-22

    Glycoproteins expressed in the human body can experience modifications as result of pathological situations. Detection of those changes can be useful as disease biomarkers. As a result of these modifications, size and/or electrical charge of the glycoprotein can be altered. Migration in capillary zone electrophoresis (CZE) is governed by the size to charge ratio of the analyte and therefore this separation technique can be used to monitor those modifications. At its turn, the alteration of the electrophoretical pattern of a given glycoprotein could be used as disease biomarker. To this aim, high repeatability for separation of a large number of peaks for a given glycoprotein is desirable. For prostate cancer, new markers are needed to decrease the high number of false positive results provided by the biomarkers currently used in clinics. In this sense, CZE methods for analysis of the several prostate specific antigen (PSA) peaks which this glycoprotein exhibit, called isoforms and containing one or more glycoforms, could be useful to study the PSA pattern as prostate cancer marker. In this study two complementary strategies to achieve both lot-to-lot capillary repeatability and high resolution of a large number of PSA isoforms are developed. Better performance and precision have been obtained for capillaries conditioned with HCl than for those conditioned with NaOH. Optimization of the background electrolyte (BGE) pH value to 8.0 and inclusion of 3M urea on its composition were the two factors of highest impact for enhancing resolution of the highest number of PSA peaks. Under the optimized conditions for capillary conditioning and BGE pH and composition, long-term resolution of 10 isoforms of PSA was achieved. Inter-day (n=3) %RSD was 0.55 for the ratio tm/tEOF, 1.15 for μeff, and 5.02 for % Acorr of the PSA peaks. PMID:27018191

  3. Capillary zone electrophoresis analysis and detection of mid-spectrum biological warfare agents

    Energy Technology Data Exchange (ETDEWEB)

    Boulet, C.A.; Townsley, C.

    1995-04-01

    DRE Suffield has initiated a research program to develop methods and equipment for field detection and laboratory identification of mid-spectrum agents, molecules of biological origin such as proteins, peptides and toxins. In this study, a highly efficient and reproducible capillary zone electrophoresis method was developed to separate and identify a series of nine peptides of defence interest: bradykinin, bradykinin fragment 1-5, substance P,ARG8-vasopressin, luteinizing hormone releasing hormone, bombesin, leucine enkephalin, methionine enkephalin, and oxytocin. Using a 50 micrometer x 47 cm capillary column, 22.5 kV separation voltage and a 100 mM pH 2.5 phosphate buffer, all nine peptide could separated in under 10 minutes. Three strategies, which could be used in a fully automated field detection and identification system, were demonstrated for the identification of unknown peptides: comparison of migration times, comparison of electrophoretic mobilities, and co-injection of multiple reference standards. These experiments demonstrate that a separation based analytical method such as capillary electrophoresis could form the basis of a generic detection system for mid-spectrum protein and peptide toxins.

  4. Raman spectroscopy and capillary electrophoresis applied to forensic colour inkjet printer inks analysis.

    Science.gov (United States)

    Król, Małgorzata; Karoly, Agnes; Kościelniak, Paweł

    2014-09-01

    Forensic laboratories are increasingly engaged in the examination of fraudulent documents, and what is important, in many cases these are inkjet-printed documents. That is why systematic approaches to inkjet printer inks comparison and identification have been carried out by both non-destructive and destructive methods. In this study, micro-Raman spectroscopy and capillary electrophoresis (CE) were applied to the analysis of colour inkjet printer inks. Micro-Raman spectroscopy was used to study the chemical composition of colour inks in situ on a paper surface. It helps to characterize and differentiate inkjet inks, and can be used to create a spectra database of inks taken from different cartridge brands and cartridge numbers. Capillary electrophoresis in micellar electrophoretic capillary chromatography mode was applied to separate colour and colourless components of inks, enabling group identification of those components which occur in a sufficient concentration (giving intensive peaks). Finally, on the basis of the obtained results, differentiation of the analysed inks was performed. Twenty-three samples of inkjet printer inks were examined and the discriminating power (DP) values for both presented methods were established in the routine work of experts during the result interpretation step. DP was found to be 94.0% (Raman) and 95.6% (CE) when all the analysed ink samples were taken into account, and it was 96.7% (Raman) and 98.4% (CE), when only cartridges with different index numbers were considered.

  5. Study of the Electrophoretic Behavior of Cephalosporins by Capillary Zone Electrophoresis

    Directory of Open Access Journals (Sweden)

    Gabriel Hancu

    2015-06-01

    Full Text Available Purpose: The aim of the study was the characterization of the electrophoretic behavior of cephalosporins from different generation having different structural characteristics in order to develop a rapid, simple and efficient capillary electrophoretic method for their identification and simultaneous separation from complex mixtures. Methods: Ten cephalosporin derivatives (cefaclor, cefadroxil, cefalexin, cefazolin, cefoxitin, cefuroxime, cefoperazone, cefotaxime, ceftazidime, ceftriaxone were analyzed by capillary zone electrophoresis using different background electrolyte solutions at different pH values. Electrophoretic mobilities of the analytes were calculated, the influence of the electrophoretic parameteres on the separation was established and the analytical conditions were optimized. Results: Taking into consideration their structural and chemical properties cephalosporins can be detected over a pH range between 6 and 10. The best results were obtained using a buffer solution containing 25 mM disodium hydrogenophosphate - 25 mM sodium dihydrogenophosphate, at a pH – 7.00, + 25 kV voltage at a temperature of 25 C, UV detection at 210 nm. Using the optimized analytical conditions we achieved the simultaneous baseline separation for seven cephalosporins in less then 10 minutes. Conclusion: Using the described optimized electrophoretic procedures, capillary electrophoresis can be used for the identification and determination of cephalosporins in formulated pharmaceutical products and for their separation from complex mixtures.

  6. Analysis of serotonin in brain microdialysates using capillary electrophoresis and native laser-induced fluorescence detection.

    Science.gov (United States)

    Benturquia, Nadia; Couderc, François; Sauvinet, Valérie; Orset, Cyrille; Parrot, Sandrine; Bayle, Christophe; Renaud, Bernard; Denoroy, Luc

    2005-03-01

    Serotonin or 5-hydroxytryptamine (5-HT) is a major neurotransmitter in the central nervous system. In this work, a method for analyzing 5-HT in brain microdialysis samples using a commercially available capillary electrophoresis (CE) system has been developed. A pH-mediated in-capillary preconcentration of samples was performed, and after separation by capillary zone electrophoresis, native fluorescence of 5-HT was detected by a 266 nm solid-state laser. The separation conditions for the analysis of 5-HT in standard solutions and microdialysates have been optimized, and this method has been validated on both pharmacological and analytical bases. Separation of 5-HT was performed using a 80 mmol/L citrate buffer, pH 2.5, containing 20 mmol/L hydroxypropyl-beta-cyclodextrin (HP-beta-CD) and +30 kV voltage. The detection limit was 2.5 x 10(-10) mol/L. This method allows the in vivo brain monitoring of 5-HT using a simple, accurate CE measurement in underivatized microdialysis samples.

  7. Chiral separation of benzoporphyrin derivative mono- and diacids by laser induced fluorescence-capillary electrophoresis.

    Science.gov (United States)

    Peng, Xuejun; Sternberg, Ethan; Dolphin, David

    2002-01-01

    A method for the separation of benzoporphyrin derivative mono- and diacid (BPDMA, BPDDA) enantiomers by laser induced fluorescence-capillary electrophoresis (LIF-CE) has been developed. By using 300 mM borate buffer, pH 9.2, 25 mM sodium cholate and 10% acetronitrile as electrolyte, +10 kV electrokinetic sampling injection of 2 s and an applied +20 kV voltage across the ends of a 37 cm capillary (30 cm to the detector, 50 microm ID), all six BPD stereoisomers were baseline-separated within 20 min. Formation constants, free electrophoretic and complexation mobilities with borate and cholate were determined based on dynamic complexation capillary electrophoresis theory. The BPD enantiomers can be quantitatively determined in the range of 10(-2)-10(-5) mg mL(-1). The correlation coefficients (r2) of the least-squares linear regression analysis of the BPD enantiomers are in the range of 0.9914-0.9997. Their limits of detection are 2.18-3.5 x 10(-3) mg mL(-1). The relative standard deviations for the separation were 2.90-4.64% (n = 10). In comparison with high-performance liquid chromatography (HPLC), CE has better resolution and efficiency. This separation method was successfully applied to the BPD enantiomers obtained from a matrix of bovine serum and from liposomally formulated material as well as from studies with rat, dog and human microsomes. PMID:11824627

  8. Two-dimensional separation of ionic species by hyphenation of capillary ion chromatography × capillary electrophoresis-mass spectrometry.

    Science.gov (United States)

    Beutner, Andrea; Kochmann, Sven; Mark, Jonas Josef Peter; Matysik, Frank-Michael

    2015-03-17

    The separation of complex mixtures such as biological or environmental samples requires high peak capacities, which cannot be established with a single separation technique. Therefore, multidimensional systems are in demand. In this work, we present the hyphenation of the two most important (orthogonal) techniques in ion analysis, namely, ion chromatography (IC) and capillary electrophoresis (CE), in combination with mass spectrometry. A modulator was developed ensuring a well-controlled coupling of IC and CE separations. Proof-of-concept measurements were performed using a model system consisting of nucleotides and cyclic nucleotides. The data are presented in a multidimensional contour plot. Analyte stacking in the CE separation could be exploited on the basis of the fact that the suppressed IC effluent is pure water.

  9. Capillary electrophoresis of proteins in buffers containing high concentrations of zwitterionic salts.

    Science.gov (United States)

    Bushey, M M; Jorgenson, J W

    1989-10-20

    A method for improving protein separations in capillary zone electrophoresis utilizing high concentrations of zwitterionic buffer additives was examined. Lysozyme and alpha-chymotrypsinogen A were used as test proteins in untreated fused-silica capillaries in buffers of pH ca. 7.0 and 9.0 The zwitterion-containing buffers were compared with buffers containing high ionic salt concentrations and a buffer containing a combination of high ionic salt and high zwitterion concentrations. Over 100,000 theoretical plates were obtained in less than 30 min. for both test proteins in a pH 7 buffer containing both trimethylglycine and potassium sulfate. The advantages and disadvantages of this technique compared with those of other methods used to prevent protein adsorption are discussed. PMID:2592485

  10. Affinity capillary electrophoresis method for investigation of bile salts complexation with sulfobutyl ether-ß-cyclodextrin

    DEFF Research Database (Denmark)

    Østergaard, Jesper; Jensen, Henrik; Holm, Rene

    2012-01-01

    Sulfobutyl ether-ß-cyclodextrin (SBEßCD) is utilized in preformulation and drug formulation as an excipient for solubilization of drugs with poor aqueous solubility. Approximately seven negative charges of SBEßCD play a role with respect to solubilization and complexation, but also have...... an influence on the ionic strength of the background electrolyte when the cyclodextrin is used in capillary electrophoresis. Mobility-shift affinity capillary methods for investigation of the complexation of taurocholate and taurochenodeoxycholate with the negatively charged cyclodextrin derivative applying...... for the bile salts were in the same range as those previously reported for the interaction with neutral ß-cyclodextrins derivatives, i.e. the positions of the negative charges on SBEßCD and the bile salts within the complex did not lead to significant electrostatic repulsion....

  11. Inter-instrumental method transfer of chiral capillary electrophoretic methods using robustness test information.

    Science.gov (United States)

    De Cock, Bart; Borsuk, Agnieszka; Dejaegher, Bieke; Stiens, Johan; Mangelings, Debby; Vander Heyden, Yvan

    2014-08-01

    Capillary electrophoresis (CE) is an electrodriven separation technique that is often used for the separation of chiral molecules. Advantages of CE are its flexibility, low cost and efficiency. On the other hand, the precision and transfer of CE methods are well-known problems of the technique. Reasons for the more complicated method transfer are the more diverse instrumental differences, such as total capillary lengths and capillary cooling systems; and the higher response variability of CE methods compared to other techniques, such as liquid chromatography (HPLC). Therefore, a larger systematic change in peak resolutions, migration times and peak areas, with a loss of separation and efficiency may be seen when a CE method is transferred to another laboratory or another type of instrument. A swift and successful method transfer is required because development and routine use of analytical methods are usually not performed in the same laboratory and/or on the same type of equipment. The aim of our study was to develop transfer rules to facilitate CE method transfers between different laboratories and instruments. In our case study, three β-blockers were chirally separated and inter-instrumental transfers were performed. The first step of our study was to optimise the precision of the chiral CE method. Next, a robustness test was performed to identify the instrumental and experimental parameters that were most influencing the considered responses. The precision- and the robustness study results were used to adapt instrumental and/or method settings to improve the transfer between different instruments. Finally, the comparison of adapted and non-adapted transfers allowed deriving some rules to facilitate CE method transfers.

  12. New electrolyte systems for capillary zone electrophoresis of metal cations and non-ionic organic compounds

    Energy Technology Data Exchange (ETDEWEB)

    Shi, Y.

    1995-06-19

    Excellent separations of metal ions can be obtained very quickly by capillary electrophoresis provided a weak complexing reagent is incorporated into the electrolyte to alter the effective mobilities of the sample ions. Indirect photometric detection is possible by also adding a UV-sensitive ion to the electrolyte. Separations are described using phthalate, tartrate, lactate or hydroxyisobutyrate as the complexing reagent. A separation of twenty-seven metal ions was achieved in only 6 min using a lactate system. A mechanism for the separation of lanthanides is proposed for the hydroxyisobutyrate system.

  13. Rapid determination of salbutamol in pharmaceutical preparations by chiral capillary electrophoresis

    OpenAIRE

    Ekiert, Ela; García Ruiz, Carmen; García, María A.; Marina Alegre, María Luisa

    2003-01-01

    A fast and simple method of chiral capillary electrophoresis (CE) has been applied to the analysis of salbutamol in different pharmaceutical preparations. Using of a 25 mM acetate buffer (pH 5.0), containing 13.1 mg/mL carboxymethyl- -cyclodextrin (CM- - CD), an applied voltage of 20 kV and a temperature of 25 C, the enantiomers of salbutamol could be separated in about 2 min. Three different pharmaceutical preparations (two syrups, one oral solution, and two kind of tablets) containing a ...

  14. Studies of Active Ingredients in Cough Syrup by Capillary Zone Electrophoresis with Amperometric Detection

    Institute of Scientific and Technical Information of China (English)

    ZHOU Tian-shu; WANG Ai-fang; WU Fang; SHI Guo-yue; FANG Yu-zhi

    2003-01-01

    The present paper covers a simple, reliable and reproducible method, based on capillary zone electrophoresis(CZE) with amperometric detection(AD), for the separation and the determination of ephedrine hydrochloride, promethazine hydrochloride and codeine phosphate. Under the optimal conditions, the three analytes were base-line separated completely within 16 min. Good linear relationships between the peak heights and the concentrations of the three analytes were obtained with the correlation coefficients better than 0.9993. The method was directly applied to the determination of the active ingredients in pharmaceutical preparations and the assay results were satisfactory.

  15. Study on Rhizoma Chuanxiong based on capillary electrophoresis with amperometric detection

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    A high-performance capillary electrophoresis with amperometric detection(CE-AD) method has been developed for the analysis of seven bioactive ingredients,namely ferulic acid(FA),vanillin,vanillic acid,p-hydroxybenzoic acid,caffeic acid,gallic acid and protocatechuic acid,in Rhizoma Chuanxiong.The effects of several factors such as the acidity and concentration of running buffer,the separation voltage,the applied potential to working electrode and the injection time were investigated.Under the optimum con...

  16. CAPILLARY ELECTROPHORESIS METHOD FOR THE DETERMINATION OF LACTULOSE AND MANNITOL RATIO FOR ESTIMATING INTESTINAL PERMEABILITY

    OpenAIRE

    E. TÜRKÖZ ACAR1, D. ÖZER ÜNAL2,*, M. KOÇ3, E. TÜRKÖZ ACAR1, D. ÖZER ÜNAL2,*, M.

    2014-01-01

    SUMMARYIn this study, a simple, rapid and accurate capillary electrophoresis method has been developed for the determination of lactulose and mannitol in urine for estimating the intestinal permeability. The urine samples were prepared to analyze by using solid phase extraction. For analysis a background electrolyte (BGE) consisted of 6 mmol/L sorbic acid, 1.25mmol/L hexadecyltrimethylammoniumbromide (HDTMA), 50 mmol/L LiOH (pH 12.5) was used. Separation was performed at -5 kV over 20 min wit...

  17. Use of Capillary Electrophoresis in the Study of Interaction between dsDNA and Drug Molecules

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Two 17-mer dsDNA with different sequence characteristics were designed to investigate the binding characteristics of berberine, an anticancer drug with uncertain binding mode, and Hoechst 33258, a model DNA minor groove binder, with dsDNA, respectively by the capillary zone electrophoresis (CZE). Kenndler model analysis revealed that Hoechst 33258 exhibited intermediate affinity with dsDNA, while there was only low affinity and some weak binding preference for AATT-containing to GGCC-containing dsDNA for berberine.

  18. Optimized Separation of Pharmacologically Active Xanthones from Secuidaca inappendiculata by Capillary Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    TaoBO; XueDongYANG; 等

    2002-01-01

    A capillary electrophoresis (CE) method has been firstly used for the separation of the therapeutically important xanthones from Securidaca inappendiculata. The separation of the nine xanthones was systematically optimized with respect to pH, concentration of running buffers,addition of sulfated β-CD,applied voltage and column temperature.Baseline separation was achieved for the nine xanthones in less than 15 minutes using a background electrolyte consisting of 200 mmol/L borate (pH9.5) and 10 mmol/L sulfated β-CD.

  19. Optimized Separation of Pharmacologically Active Xanthones from Securidaca inappendiculata by Capillary Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    A capillary electrophoresis (CE) method has been firstly used for the separation of the therapeutically important xanthones from Securidaca inappendiculata. The separation of the nine xanthones was systematically optimized with respect to pH, concentration of running buffers, addition of sulfated b-CD, applied voltage and column temperature. Baseline separation was achieved for the nine xanthones in less than 15 minutes using a background electrolyte consisting of 200 mmol/L borate (pH 9.5) and 10 mmol/L sulfated b-CD.

  20. Determination of diastereoisomeric alkaloids in urine by capillary electrophoresis with electrochemiluminescence detection

    Institute of Scientific and Technical Information of China (English)

    Yan Ming Liu; Long Fei Peng; Lin Mei; Li Juan Liu

    2011-01-01

    A new and simple capillary electrophoresis with electrochemiluminescence detection was developed for the separation and the quantification of a pair of diastereoisomeric alkaloids (ephedrine and pseudoephedrine). The limits of detection (S/N = 3) were 4.5 × 10-8 mol/L for ephedrine and 5.2×10-8 mol/L for pseudoephedrine, respectively. The RSDs of migration time and peak area were less than 1.3 and 2.5% (n = 5), respectively. The applicability of the propose method was illustrated in the determination of ephedrine and pseudoephedrine in human urine, ephedrine in nasal drops, and the monitoring of pharmacokinetics for pseudoephedrine.

  1. Capillary electrophoresis with inhibited electrochemiluminescent detection for the trace analysis of epinephrine and dopamine

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    In this paper,a novel electrochemiluminescent (ECL) detection approach was developed for highly sensitive detection of ECL inhibitors based on the ECL inhibition of Ru(bpy)32+/2-(Dibutylamino)ethanol (DBAE) system. A microfluidic ECL detection cell was fabricated to couple with the capillary electrophoresis system,the electrochemical system and the postcolumn injection system. Both Ru(bpy)32+ and DBAE solutions were injected directly to the working electrode surface by a micro-infusion system to obtain a hi...

  2. A new injection method for soil nutrient analysis in capillary electrophoresis

    Science.gov (United States)

    Smolka, M.; Puchberger-Enengl, D.; Bipoun, M.; Fercher, G.; Klasa, A.; Krutzler, C.; Keplinger, F.; Vellekoop, M. J.

    2013-05-01

    We present a new method for the direct injection of liquid sample into a capillary electrophoresis (CE) device. Instead of a double-T injection mechanism, a single inlet provided with a membrane filter is used. From a reservoir on top of this inlet, the liquid directly enters the separation channel through the membrane. The driving force is a short electrical pulse. This avoids an additional sample channel, so that the chip needs only three microfluidic connects and no mechanical sample pumping is demanded. The high injection reproducibility and the comparatively simple setup open up the way for mobile application of soil analysis.

  3. Investigation of Melamine Presence in Canned Tuna Fish by Capillary Zone Electrophoresis Method

    OpenAIRE

    Er Demirhan, Buket; Demirhan, Burak; Yarımkaya Baş, Sezen; Yentür, Gülderen; Bayhan Öktem, Aysel

    2015-01-01

    Melamine is widely used as a chemical in food industry and may lead to kidney damage. The aim of this study was to determine melamine and pH value of 80 canned tuna fish samples of four different brands (A, B, C, D) sold in Ankara, Turkey. Quantitative determination of melamine in canned tuna fish samples was carried out by capillary zone electrophoresis with diode array detector (CZE-DAD). The limits of detection and quantitation for melamine were found to be 0.21 mg kg-1and 0.68 mg kg-1, re...

  4. Capillary electrophoresis with direct chemiluminescence detection for the analysis of catecholamines in human urine

    Institute of Scientific and Technical Information of China (English)

    Cheng Quan Wang; Hui Wang; Yan Ming Liu

    2007-01-01

    A rapid and sensitive method for the analysis of three catecholamines by capillary electrophoresis (CE) with direct chemiluminescence (CL) detection is described. The detection limits (S/N = 3) were 1.3 × 10-8 g/mL for isoprenaline,1.0 × 10-8 g/mL for epinephrine and 2.8 × 10-8 g/mL for dopamine. The proposed method was successfully applied to the analysis of catecholamines in urine samples of cigarette smokers and nonsmokers. The results showed that there is a close relation between the release of dopamine in human body fluids and cigarette smoking/nonsmoking.

  5. Separation of chiral drugs with β-CD phosphate by capillary electrophoresis

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    β -Cyclodextrin phosphate (β -CD-phosphate) was used as a selector for separating chiral drugs by capillary electrophoresis (CE). A solution comprising of 120 mmol/L Britton-Robinson buffer (BRB) containing 10 mmol/L β -CD phosphate with the pH adjusted to 7.0 was used as the background electrolyte (BGE), and a small amount of analyte was injected (600v/1s). Triethylamine, diethylamine, triethanolamine, diethanolamine, Tris added as modifier were compared. Isoprenaline, methoxamine, oxprenolol, practolol were successfully resolved.

  6. Self-assembly of cellulose nanoparticles as electrolyte additive for capillary electrophoresis separation.

    Science.gov (United States)

    Huang, Dihui; Yang, Qin; Jin, Shanxia; Deng, Qianchun; Zhou, Ping

    2014-11-01

    In this work, a new cellulose derivative, octadecyl modified quaternized cellulose (ODMQC), was synthesized and used as additive in the background electrolyte for capillary electrophoresis. The derivative bearing hydrophobic groups and hydrophilic groups can self-assemble into a stable nano-scaled micelle structure in aqueous solution. When ODMQC was added in running buffer, the capillaries were shown to generate applicable anodal EOF over the investigated range of pH 3.0-12.0. Due to the lack of UV active groups, the ODMQC did not disturb the UV detection. It is shown that ODMQC-added capillaries allow the separation of basic proteins by reducing their adsorption onto the capillary wall. Also, the addition of ODMQC provides adequate separation of aromatic acids with low pKa values and improved separation of sulfa drugs. Moreover, it is demonstrated that the addition of ODMQC can incorporate an additional reversed-phase mechanism that improves the separation of neutral analytes. PMID:25262028

  7. A new post-column reactor-laser induced fluorescence detector for capillary electrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Zhang Liling

    1996-01-02

    Capillary zone electrophoresis (CZE), a powerful separation method based on the differential migration of charged species under the influence of an electric field, has been widely used for separations covering from small ions to big biomolecules. Chapter 1 describes the method, then discusses detection of the separated analytes by laser induced fluorescence and by chemical derivatization, and the use of O-phthaldialdehyde (OPA) as a post-column reagent. Chapter 2 describes a post-column reactor which uses two narrow bore capillaries connected coaxially. This reactor differs from other coaxial reactors in terms of capillary dimensions, reagent flow control, ease of construction and most importantly, better limits of detection. The derivatization reagent is electroosmotically driven into the reaction capillary and the reagent flow rate is independently controlled by a high voltage power supply. Amino acids, amines and proteins, derivatized by OPA/2-mercaptoethanol using this post-column reactor coupled with LIF detection, show low attomole mass limits of detection, and for the first time, the authors demonstrate single cell capability with a post-column derivatization scheme. The single cell capability shows that this reactor could find applications in assaying non-fluorescent or electrochemically inactive components in individual biological cells in the future.

  8. Sensitive Method for Enantioseparation of Rivastigmine with Highly Sulfated Cyclodextrin as Chiral Selector by Capillary Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    WANG Zhao-Yan; XU Xing-Xiang; HU Zhi-De; KANG Jing-Wu

    2006-01-01

    A sensitive method for enantioseparation of a basic drug rivastigmine and determination of its optical impurity by capillary electrophoresis with highly sulfatedβ-cyclodextrin (HS-β-CD) as the chiral selector is described. In general, enantioseparation of basic chiral compounds is carried out in acidic condition (pH 2.5) to prevent analytes from adsorption on the capillary wall. However, in the case of rivastigmine, the detection sensitivity was too limited to determine the optical impurity of S-rivastigmine lower than 1% when buffer pH was 2.5. It was found that the detection sensitivity was improved 1.6 times just by raising the buffer pH value from 2.5 to 5.8. The poor column efficiency due to the adsorption of the analytes on the capillary wall was resolved by using dynamical coating of the capillary wall with the linear polyacrylamide solution. The experimental parameters such as the concentration of HS-β-CD, buffer pH and buffer ionic strength were optimized, respectively. The method was validated in terms of repeatability, linearity, limit of detection (LOD) and limit of quantitation (LOQ). Using the present method, the optical purity of nonracemic rivastigmine with the enantiomeric excess (ee) value of 99.14% was determined.

  9. Laser ablation construction of on-column reagent addition devices for capillary electrophoresis.

    Science.gov (United States)

    Rezenom, Yohannes H; Lancaster, Joseph M; Pittman, Jason L; Gilman, S Douglass

    2002-04-01

    A simple and reproducible technique for constructing perfectly aligned gaps in fused-silica capillaries has been developed for postcolumn reagent addition with capillary electrophoresis. This technique uses laser ablation with the second harmonic of a Nd:YAG laser (532 nm) at 13.5 mJ/pulse and a repetition rate of 15 Hz to create these gaps. A capillary is glued to a microscope slide and positioned at the focal point of a cylindrical lens using the focused beam from a laser pointer as a reference. Gaps of 14.0 +/- 2.2 microm (n = 33) at the bore of the capillary are produced with a success rate of 94% by ablation with 400 pulses. This simple method of gap construction requires no micromanipulation under a microscope, hydrofluoric acid etching, or use of column fittings. These structures have been used for reagent addition for postcolumn derivatization with laser-induced fluorescence detection and have been tested for the separation of proteins and amino acids. Detection limits of 6 x 10(-7) and 1 x 10(-8) M have been obtained for glycine and tranferrin, respectively. Separation efficiencies obtained using these gap reactors range from 38,000 to 213,000 theoretical plates. PMID:12043598

  10. Determination of vanillin in vanilla perfumes and air by capillary electrophoresis.

    Science.gov (United States)

    Minematsu, Saaya; Xuan, Guang-Shan; Wu, Xing-Zheng

    2013-12-01

    The present study investigated capillary electrophoretic detection of vanillin in vanilla perfume and air. An UV-absorbance detector was used in a home-made capillary electrophoretic instrument. A fused silica capillary (outer diameter: 364 μm, inner diameter: 50 μm) was used as a separation capillary, and a high electric voltage (20 kV) was applied across the two ends of the capillary. Total length of the capillary was 70 cm, and the effective length was 55 cm. Experimental results showed that the vanillin peak was detected at about 600, 450, and 500 seconds when pH of running buffers in CE were 7.2, 9.3, and 11.5, respectively. The peak area of vanillin was proportional to its concentration in the range of 0-10(-2) mol/L. The detection limit was about 10(-5) mol/L. Vanillin concentration in a 1% vanilla perfume sample was determined to be about 3×10(-4) mol/L, agreed well with that obtained by a HPLC method. Furthermore, determination of vanillin in air by combination of CE and active carbon adsorption method was investigated. PMID:25078845

  11. Determination of vanillin in vanilla perfumes and air by capillary electrophoresis.

    Science.gov (United States)

    Minematsu, Saaya; Xuan, Guang-Shan; Wu, Xing-Zheng

    2013-12-01

    The present study investigated capillary electrophoretic detection of vanillin in vanilla perfume and air. An UV-absorbance detector was used in a home-made capillary electrophoretic instrument. A fused silica capillary (outer diameter: 364 μm, inner diameter: 50 μm) was used as a separation capillary, and a high electric voltage (20 kV) was applied across the two ends of the capillary. Total length of the capillary was 70 cm, and the effective length was 55 cm. Experimental results showed that the vanillin peak was detected at about 600, 450, and 500 seconds when pH of running buffers in CE were 7.2, 9.3, and 11.5, respectively. The peak area of vanillin was proportional to its concentration in the range of 0-10(-2) mol/L. The detection limit was about 10(-5) mol/L. Vanillin concentration in a 1% vanilla perfume sample was determined to be about 3×10(-4) mol/L, agreed well with that obtained by a HPLC method. Furthermore, determination of vanillin in air by combination of CE and active carbon adsorption method was investigated.

  12. Capillary electrophoresis for the assay of fixed-dose combination tablets of artesunate and amodiaquine

    Directory of Open Access Journals (Sweden)

    Amin N’Cho

    2012-05-01

    Full Text Available Abstract Background Quality control of drugs in formulations is still a major challenge in developing countries. For the quality control of artesunate and amodiaquine tablets in fixed-dose combination, only liquid chromatographic methods have been proposed in the literature. There are no capillary electrophoretic methods reported for the determination of these active substances, although this technique presents several advantages over liquid chromatography (long lifetime, low price of the capillary, low volumes of electrolyte consumption in addition to simplicity. In this paper, a reliable capillary electrophoresis method has been developed and validated for the quality control of these drugs in commercial fixed-dose combination tablets. Methods Artesunate and amodiaquine hydrochloride in bilayer tablets were determined by micellar electrokinetic capillary chromatography (MEKC. Analytes were extracted from tablets by sonication with a solvent mixture phosphate buffer pH 7.0-acetonitrile containing benzoic acid as internal standard. Separation was carried out on Beckman capillary electrophoresis system equipped with fused silica capillary, 30 cm long (20 cm to detector × 50 μm internal diameter, using a 25 mM borate buffer pH 9.2 containing 30 mM sodium dodecyl sulfate as background electrolyte, a 500 V cm−1 electric field and a detection wavelength of 214 nm. Results Artesunate, amodiaquine and benzoic acid were separated in 6 min. The method was found to be reliable with respect to specificity,linearity of the calibration line (r2 > 0.995, recovery from synthetic tablets (in the range 98–102%, repeatability (RSD 2–3%, n = 7 analytical procedures. Application to four batches of commercial formulations with different dosages gave content in good agreement with the declared content. Conclusion The MEKC method proposed is reliable for the determination of artesunate and amodiaquine hydrochloride in fixed

  13. A fluorescence detection scheme for capillary electrophoresis of N-methylcarbamates with on-column thermal decomposition and derivatization

    Science.gov (United States)

    Wu; Lee; Li

    2000-04-01

    This paper describes a fluorescence detection method for N-methylcarbamate (NMC) pesticides in micellar electrokinetic chromatography (MEKC) separation. Fulfillment of the fluorescence detection hinged on the discovery that quaternary ammonium surfactants (particularly cetyltrimethylammonium bromide, CTAB), besides serving as hydrophobic pseudophases in MEKC, are also capable of catalyzing the thermal decomposition of NMCs to liberate methylamine. Thus, a multifunctional MEKC medium consisting of borate buffer, CTAB, and derivatizing components (o-phthaldialdehyde/2-mercaptoethanol) was formulated, which allowed first normal MEKC separation, subsequent thermal decomposition, and finally in situ derivatization of NMCs. With careful optimization of the operation conditions, fluorescence detection of 10 NMC compounds was achieved, with column efficiencies typically higher than 50,000 and detection limits better than 0.5 ppm. The present work represents an unprecedented effort in capillary electrophoresis (CE), in which an intact capillary was consecutively utilized as chambers for separation, decomposition, derivatization, and detection, without involving any interfacing features. The success in the implementation of such a detection system resulted in strikingly simple instrumentation as compared with the traditional postcolumn fluorescence determination of NMCs by reversed-phase HPLC. Similar protocols should be workable in the determination of a wide range of pesticides and pharmaceuticals in CE formats. PMID:10763238

  14. Rapid identification and quantitation for oral bacteria based on short-end capillary electrophoresis.

    Science.gov (United States)

    Chen, Jin; Ni, Yi; Liu, Chenchen; Yamaguchi, Yoshinori; Chen, Qinmiao; Sekine, Shinichi; Zhu, Xifang; Dou, Xiaoming

    2016-11-01

    High-speed capillary electrophoresis (HSCE) is a promising technology applied in ultra-rapid and high-performance analysis of biomolecules (such as nucleic acids, protein). In present study, the short-end capillary electrophoresis coupled with one novel space domain internal standard method (SDIS) was employed for the rapid and simultaneous analysis of specific genes from three oral bacteria (Porphyromonas gingivalis (P.g), Treponema denticola (T.d) and Tannerela forsythia (T.f)). The reliability, reproducibility and accuracy properties of above mentioned SDIS method were investigated in detail. The results showed the target gene fragments of P.g, T.d and T.f could be precisely, fast identified and quantitated within 95s via present short-end CE system. The analyte concentration and the ratio of space domain signals (between target sample and internal standard sample) featured a well linear relationship calculated via SDIS method. And the correlation coefficients R(2) and detection limits for P.g, T.d, T.f genes were 0.9855, 0.9896, 0.9969 and 0.077, 0.114 and 0.098ng/μl, respectively. PMID:27591633

  15. Determination of artificial sweeteners by capillary electrophoresis with contactless conductivity detection optimized by hydrodynamic pumping.

    Science.gov (United States)

    Stojkovic, Marko; Mai, Thanh Duc; Hauser, Peter C

    2013-07-17

    The common sweeteners aspartame, cyclamate, saccharin and acesulfame K were determined by capillary electrophoresis with contactless conductivity detection. In order to obtain the best compromise between separation efficiency and analysis time hydrodynamic pumping was imposed during the electrophoresis run employing a sequential injection manifold based on a syringe pump. Band broadening was avoided by using capillaries of a narrow 10 μm internal diameter. The analyses were carried out in an aqueous running buffer consisting of 150 mM 2-(cyclohexylamino)ethanesulfonic acid and 400 mM tris(hydroxymethyl)aminomethane at pH 9.1 in order to render all analytes in the fully deprotonated anionic form. The use of surface modification to eliminate or reverse the electroosmotic flow was not necessary due to the superimposed bulk flow. The use of hydrodynamic pumping allowed easy optimization, either for fast separations (80s) or low detection limits (6.5 μmol L(-1), 5.0 μmol L(-1), 4.0 μmol L(-1) and 3.8 μmol L(-1) for aspartame, cyclamate, saccharin and acesulfame K respectively, at a separation time of 190 s). The conditions for fast separations not only led to higher limits of detection but also to a narrower dynamic range. However, the settings can be changed readily between separations if needed. The four compounds were determined successfully in food samples. PMID:23830447

  16. Angiotensin-converting enzyme inhibition studies by natural leech inhibitors by capillary electrophoresis and competition assay.

    Science.gov (United States)

    Deloffre, Laurence; Sautiere, Pierre-Eric; Huybrechts, Roger; Hens, Korneel; Vieau, Didier; Salzet, Michel

    2004-06-01

    A protocol to follow the processing of angiotensin I into angiotensin II by rabbit angiotensin-converting enzyme (ACE) and its inhibition by a novel natural antagonist, the leech osmoregulator factor (LORF) using capillary zonal electrophoresis is described. The experiment was carried out using the Beckman PACE system and steps were taken to determine (a) the migration profiles of angiotensin and its yielded peptides, (b) the minimal amount of angiotensin II detected, (c) the use of different electrolytes and (d) the concentration of inhibitor. We demonstrated that LORF (IPEPYVWD), a neuropeptide previously found in leech brain, is able to inhibit rabbit ACE with an IC(50) of 19.8 micro m. Interestingly, its cleavage product, IPEP exhibits an IC(50) of 11.5 micro m. A competition assay using p-benzoylglycylglycylglycine and insect ACE established that LORF and IPEP fragments are natural inhibitors for invertebrate ACE. Fifty-four percent of insect ACE activity is inhibited with 50 micro m IPEP and 35% inhibition with LORF (25 mm). Extending the peptide at both N- and C-terminus (GWEIPEPYVWDES) and the cleavage of IPEP in IP abolished the inhibitory activity of both peptides. Immunocytochemical data obtained with antisera raised against LORF and leech ACE showed a colocalization between the enzyme and its inhibitor in the same neurons. These results showed that capillary zonal electrophoresis is a useful technique for following enzymatic processes with small amounts of products and constitutes the first evidence of a natural ACE inhibitor in invertebrates.

  17. Genomic DNA detection using cycling probe technology and capillary gel electrophoresis with laser-induced fluorescence.

    Science.gov (United States)

    Dickinson Laing, Terrina; Mah, David C W; Poirier, Robert T; Bekkaoui, Faouzi; Lee, William E; Bader, Douglas E

    2004-10-01

    Cycling probe technology (CPT) is an isothermal DNA analysis method that has been shown to be useful for identifying genetic markers of antibiotic-resistant bacteria in clinical settings. CPT assays have previously employed several assay methods that include polyacrylamide gel electrophoresis and magnetic beads for separations and radioisotopic and colorimetric detection for detection. Capillary gel electrophoresis with laser-induced fluorescence (CGE-LIF) is an alternative separation and detection method that offers attributes such as low sample consumption, short analysis times, no radiation hazards and potential for high throughput. We report on the development of a CGE-LIF CPT assay for genomic DNA from Erwinia herbicola and the comparison of this assay with a conventional 32p radioisotopic PAGE CPT assay. Separation and detection of intact and cleaved fluorescein-labeled CPT probe molecules by CGE-LIF was achieved in under 4 min through a gel-filled capillary (7 cm separation length to detector). Total time, from setup to detection, was about 1 h for the complete assay versus several hours (3-12 h) for the radioisotopic PAGE CPT assay. Similar detection limits of 10(5)-10(6) copies of genomic target DNA were observed with each assay method. This study demonstrated that CGE-LIF CPT is a suitable analysis method for the detection of genomic DNA sequences. PMID:15356906

  18. Protonated diamines as anion-binding agents and their utility in capillary electrophoresis separations.

    Science.gov (United States)

    Noblitt, Scott D; Speights, Rachel M; Henry, Charles S

    2011-11-01

    Capillary zone electrophoresis is a proven method for separating small ions because of the inherent charge and differences in mobility of these analytes. Despite its resolving power, CZE can be insufficient for separating ions with similar mobilities. One remedy is to modify mobilities via the addition of background electrolyte complexation agents. However, this approach is not straightforward for inorganic anions, which lack complexation options. To address this shortfall, the diprotonated diamine moiety was investigated for complexation of dianions. Dicationic diamines significantly complexed dianions, and this interaction was not purely electrostatic in nature because affinities varied with dianion identity. Aqueous association constants were measured with affinity capillary electrophoresis (ACE) and found to be similar in magnitude but different in selectivity to those of dianions with magnesium ion. Binding was also investigated for zwitterionic buffers containing the protonated diamine moiety. Zwitterions exhibited binding constants as high as 18 M(-1) (30-mM ionic strength). This work discusses the observed binding constants and their potential usefulness in CZE separations of inorganic anions. Also covered are improvements to ACE methodology and an evaluation of some of the assumptions employed. PMID:22002837

  19. Simultaneous determination of phenylethanoid glycosides and aglycones by capillary zone electrophoresis with running buffer modifier.

    Science.gov (United States)

    Dong, Shuqing; Gao, Ruibin; Yang, Yan; Guo, Mei; Ni, Jingman; Zhao, Liang

    2014-03-15

    Although the separation efficiency of capillary electrophoresis (CE) is much higher than that of other chromatographic methods, it is sometimes difficult to adequately separate the complex ingredients in biological samples. This article describes how one effective and simple way to develop the separation efficiency in CE is to add some modifiers to the running buffer. The suitable running buffer modifier β-cyclodextrin (β-CD) was explored to fast and completely separate four phenylethanoid glycosides and aglycones (homovanillyl alcohol, hydroxytyrosol, 3,4-dimethoxycinnamic acid, and caffeic acid) in Lamiophlomis rotata (Lr) and Cistanche by capillary zone electrophoresis with ultraviolet (UV) detection. It was found that when β-CD was used as running buffer modifier, a baseline separation of the four analytes could be accomplished in less than 20 min and the detection limits were as low as 10(-3) mg L(-1). Other factors affecting the CE separation, such as working potential, pH value and ionic strength of running buffer, separation voltage, and sample injection time, were investigated extensively. Under the optimal conditions, a successful practical application on the determination of Lr and Cistanche samples confirmed the validity and practicability of this method.

  20. Congophilicity (Congo red affinity) of different beta2-microglobulin conformations characterized by dye affinity capillary electrophoresis

    DEFF Research Database (Denmark)

    Heegaard, N H; Sen, J W; Nissen, Mogens Holst

    2000-01-01

    The amyloidogenic protein beta-microglobulin was characterized by affinity capillary electrophoresis (CE). CE could separate conformational variants of beta2-microglobulin and with the amyloid-specific dye Congo red as a buffer additive it was possible to measure different Congo red-affinities of......The amyloidogenic protein beta-microglobulin was characterized by affinity capillary electrophoresis (CE). CE could separate conformational variants of beta2-microglobulin and with the amyloid-specific dye Congo red as a buffer additive it was possible to measure different Congo red......-affinities of native and abnormally folded beta2-microglobulin. We find that native beta2-microglobulin has an intermediate affinity for Congo red at pH 7.3 and that binding involves electrostatic interactions. The conformational variant of beta2-microglobulin that appears in acetonitrile solutions binds Congo red...... more strongly. Affinity CE using Congo red as a buffer additive is a new, simple, fast, and quantitative micromethod for the characterization of soluble conformational intermediates of amyloidogenic proteins....

  1. Monitoring antigenic protein integrity during glycoconjugate vaccine synthesis using capillary electrophoresis-mass spectrometry.

    Science.gov (United States)

    Tengattini, Sara; Domínguez-Vega, Elena; Temporini, Caterina; Terreni, Marco; Somsen, Govert W

    2016-09-01

    A capillary electrophoresis-mass spectrometry (CE-MS) method was developed for the characterization and integrity assessment of the Mycobacterium tuberculosis (MTB) antigens TB10.4 and Ag85B and their chemically produced glycoconjugates, which are glycovaccine candidates against tuberculosis (TB). In order to prevent protein adsorption to the inner capillary wall and to achieve efficient separation of the antigen proteoforms, a polyionic multilayer coating of polybrene-dextran sulfate-polybrene (PB-DS-PB) was used in combination with 1.5 M acetic acid as background electrolyte (BGE). Coupling of CE to high-resolution time-of-flight MS was achieved by a coaxial interface employing a sheath liquid of isopropanol-water (50:50, v/v) containing 0.1 % formic acid. The MTB antigens were exposed to experimental conditions used for chemical glycosylation (but no activated saccharide was added) in order to investigate their stability during glycovaccine production. CE-MS analysis revealed the presence of several closely related degradation products, including truncated, oxidized and conformational variants, which were assigned by accurate mass. Analysis of synthesized mannose conjugates of TB10.4 and Ag85B allowed the determination of the glycoform composition of the neo-glycoproteins next to the characterization of degradation products which were shown to be partly glycoconjugated. Moreover, the selectivity of CE-MS allowed specific detection of deamidated species (protein mass change of 1.0 Da only), indicating that chemical glycosylation increased susceptibility to deamidation. Overall, the results show that CE-MS represents a useful analytical tool for the detailed characterization and optimization of neo-glycoconjugate products. Graphical Abstract Flowchart illustrating Mycobacterium tuberculosis (MTB) antigen glycosylation, glycoconjugate variant and degradation product separation by capillary electrophoresis (CE) and their characterization by intact mass

  2. Cyclodextrine Screening for the Chiral Separation of Amlodipine Enantiomers by Capillary Electrophoresis

    Directory of Open Access Journals (Sweden)

    Gabriel Hancu

    2015-03-01

    Full Text Available Purpose: Amlodipine is a long acting, dihydropyridine type calcium channel blocker frequently used in the treatment of hypertension and coronary insufficiency. The calcium channel blocking activity resides primarily in the S-amlodipine enantiomer, while R-amlodipine is a potent inhibitor of smooth muscle cell migration. Methods: In this study capillary electrophoresis was applied for the enantiomeric separation of amlodipine using different native and derivatized; neutral and charged cyclodextrines as chiral selectors. The effects of pH and composition of the background electrolyte, concentration and type of chiral selector, capillary temperature, running voltage and injection parameters have been investigated. Results: Stereoselective interactions were observed when using α-CD, β-CD, HP-β-CD, RAMEB, CM-β-CD and SBE-β-CD. Optimized separation conditions consisted on a 50 mM phosphate buffer, pH – 3.0, 20 mM RAMEB as chiral selector, + 25 kV applied voltage, 15°C temperature and UV detection at 238 nm. Using the optimized electrophoretic conditions we succeeded the chiral separation of amlodipine enantiomers in approximately 6 minute, the order of migration being R-amlodipine followed by S-amlodipine. The method was successfully applied for the determination of amlodipine enantiomers from commercially available pharmaceuticals. The linearity range, limits of detection and quantification, precision and accuracy were determined and the results obtained confirmed that the method was suitable for this purpose. Conclusion: It can be concluded that the proposed capillary electrophoresis methods can be useful for routine pharmaceutical applications with benefits of its effectivity, simplicity, short analysis time and low consumption of analytes, solvents and chiral selectors.

  3. Automated sample preparation and analysis using a sequential-injection-capillary electrophoresis (SI-CE) interface.

    Science.gov (United States)

    Kulka, Stephan; Quintás, Guillermo; Lendl, Bernhard

    2006-06-01

    A fully automated sequential-injection-capillary electrophoresis (SI-CE) system was developed using commercially available components as the syringe pump, the selection and injection valves and the high voltage power supply. The interface connecting the SI with the CE unit consisted of two T-pieces, where the capillary was inserted in one T-piece and a Pt electrode in the other (grounded) T-piece. By pressurising the whole system using a syringe pump, hydrodynamic injection was feasible. For characterisation, the system was applied to a mixture of adenosine and adenosine monophosphate at different concentrations. The calibration curve obtained gave a detection limit of 0.5 microg g(-1) (correlation coefficient of 0.997). The reproducibility of the injection was also assessed, resulting in a RSD value (5 injections) of 5.4%. The total time of analysis, from injection, conditioning and separation to cleaning the capillary again was 15 minutes. In another application, employing the full power of the automated SIA-CE system, myoglobin was mixed directly using the flow system with different concentrations of sodium dodecyl sulfate (SDS), a known denaturing agent. The different conformations obtained in this way were analysed with the CE system and a distinct shift in migration time and decreasing of the native peak of myoglobin (Mb) could be observed. The protein samples prepared were also analysed with off-line infrared spectroscopy (IR), confirming these results. PMID:16732362

  4. Rugged gap reactor device for postcolumn fluorescence detection in capillary electrophoresis.

    Science.gov (United States)

    Wei, H; Li, S F

    1998-12-01

    In this paper, the construction and performance of a rugged device for postcolumn derivatization in capillary electrophoresis (CE) are described. The device was based on a gap design, and a gap with a very small distance (derivatizing reagents into the reaction capillary was attributable to gravity flow. The concentration of derivatizing reagents can be controlled through manipulating the electroosmotic flow in the reaction capillary and the height of the liquid levels from the derivatizing reagents to the buffer reservoirs. The device has been applied in fluorescence detection of amino acids using a mixture of o-phthaldialdehyde/2-mercaptoethanol as derivatizing reagent. Theoretical plate numbers for 11 amino acids separated in a pH 9.5 borate buffer were obtained in the order of 40 000-250 000. The detection limit for glycine (S/N = 2) was found to be 6.7 × 10(-)(7) mol/L using a commercial HPLC fluorescence detector modified for CE. Free amino acids in a wine sample were also determined. Because the device is quite stable, we believe that it can be used routinely in analytical laboratories. PMID:21644687

  5. Quality criterion to optimize separations in capillary electrophoresis: Application to the analysis of harmala alkaloids.

    Science.gov (United States)

    Tascon, Marcos; Benavente, Fernando; Castells, Cecilia B; Gagliardi, Leonardo G

    2016-08-19

    In capillary electrophoresis (CE), resolution (Rs) and selectivity (α) are criteria often used in practice to optimize separations. Nevertheless, when these and other proposed parameters are considered as an elementary criterion for optimization by mathematical maximization, certain issues and inconsistencies appear. In the present work we analyzed the pros and cons of using these parameters as elementary criteria for mathematical optimization of capillary electrophoretic separations. We characterized the requirements of an ideal criterion to qualify separations within the framework of mathematical optimizations and, accordingly, propose: -1- a new elementary criterion (t') and -2- a method to extend this elementary criterion to compose a global function that simultaneously qualifies many different aspects, also called multicriteria optimization function (MCOF). In order to demonstrate this new concept, we employed a group of six alkaloids with closely related structures (harmine, harmaline, harmol, harmalol, harmane and norharmane). On the basis of this system, we present a critical comparison between the new optimization criterion t' and the former elementary criteria. Finally, aimed at validating the proposed methods, we composed an MCOF in which the capillary-electrophoretic separation of the six model compounds is mathematically optimized as a function of pH as the unique variable. Experimental results subsequently confirmed the accuracy of the model. PMID:27443250

  6. Determination of the specific activities of methionine sulfoxide reductase A and B by capillary electrophoresis.

    Science.gov (United States)

    Uthus, Eric O

    2010-06-01

    A capillary electrophoresis (CE) method for the determination of methionine sulfoxide reductase A and methionine sulfoxide reductase B activities in mouse liver is described. The method is based on detection of the 4-(dimethylamino)azobenzene-4'-sulfonyl derivative of l-methionine (dabsyl Met), the product of the enzymatic reactions when either dabsyl l-methionine S-sulfoxide or dabsyl l-methionine R-sulfoxide is used as a substrate. The method provides baseline resolution of the substrates and, therefore, can be used to easily determine the purity of the substrates. The method is rapid ( approximately 20min sample to sample), requires no column regeneration, and uses very small amounts of buffers. Separation was performed by using a 75-mum internal diameter polyimide-coated fused silica capillary (no inside coating) with 60cm total length (50cm to the detector window). Samples were separated at 22.5kV, and the separation buffer was 25mM KH(2)PO(4) (pH 8.0) containing 0.9ml of N-lauroylsarcosine (sodium salt, 30% [w/v] solution) per 100ml of buffer. Prior to use, the capillary was conditioned with the same buffer that also contained 25mM sodium dodecyl sulfate. The CE method is compared with high-performance liquid chromatography (HPLC) as determined by comparing results from measurements of hepatic enzyme activities in mice fed either deficient or adequate selenium. PMID:20167203

  7. Determination of biogenic amines in beer and wine by capillary electrophoresis-tandem mass spectrometry.

    Science.gov (United States)

    Daniel, Daniela; Dos Santos, Vagner Bezerra; Vidal, Denis Tadeu Rajh; do Lago, Claudimir Lucio

    2015-10-16

    A capillary electrophoresis-tandem mass spectrometry (CE-MS/MS) method for the simultaneous assessment of nine biogenic amines (spermine, spermidine, putrescine, cadaverine, histamine, phenylethylamine, tryptamine, tyramine, and urocanic acid) in commercial samples of beer and wine is introduced. The samples were submitted to a simple clean-up step with poly(vinylpolypyrrolidone) followed by filtration. Electrophoretic separation in a polyvinyl alcohol (PVA)-coated capillary using 0.5 mol L(-1) acetic acid (pH 2.5) as background electrolyte and detection by electrospray-tandem mass spectrometry was employed. The range of the correlation coefficients of the calibration curves of the analyzed compounds was 0.996-0.999, and the limits of detection and limits of quantification were in the range of 1-2 μg L(-1) and 3-8 μg L(-1), respectively. The recovery values for samples spiked at three concentration levels (0.2, 0.5, and 1.0 mg L(-1)) ranged from 87 to 113% with standard deviation not greater than 5.8%. The use of a PVA-coated silica capillary allows suppressing the electroosmotic flow and, consequently, increasing of the separation efficiency. The method was successfully used to determine biogenic amines in commercial samples of beer and wine. PMID:26362807

  8. Determination of biogenic amines in beer and wine by capillary electrophoresis-tandem mass spectrometry.

    Science.gov (United States)

    Daniel, Daniela; Dos Santos, Vagner Bezerra; Vidal, Denis Tadeu Rajh; do Lago, Claudimir Lucio

    2015-10-16

    A capillary electrophoresis-tandem mass spectrometry (CE-MS/MS) method for the simultaneous assessment of nine biogenic amines (spermine, spermidine, putrescine, cadaverine, histamine, phenylethylamine, tryptamine, tyramine, and urocanic acid) in commercial samples of beer and wine is introduced. The samples were submitted to a simple clean-up step with poly(vinylpolypyrrolidone) followed by filtration. Electrophoretic separation in a polyvinyl alcohol (PVA)-coated capillary using 0.5 mol L(-1) acetic acid (pH 2.5) as background electrolyte and detection by electrospray-tandem mass spectrometry was employed. The range of the correlation coefficients of the calibration curves of the analyzed compounds was 0.996-0.999, and the limits of detection and limits of quantification were in the range of 1-2 μg L(-1) and 3-8 μg L(-1), respectively. The recovery values for samples spiked at three concentration levels (0.2, 0.5, and 1.0 mg L(-1)) ranged from 87 to 113% with standard deviation not greater than 5.8%. The use of a PVA-coated silica capillary allows suppressing the electroosmotic flow and, consequently, increasing of the separation efficiency. The method was successfully used to determine biogenic amines in commercial samples of beer and wine.

  9. The multi-concentration and two-dimensional capillary electrophoresis method for the analysis of drugs in urine samples

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    A novel method has been developed by integration of multi-concentration and two-dimensional(2D) capillary electrophoresis(CE) for simultaneous enhancement of detection sensitivity and separation power in complex samples.Capillary zone electrophoresis(CZE) was used as the first dimension separation according to mobilities,from which the effluent fractions were further analyzed by micellar electrokinetic capillary chromatography(MEKC) acting as the second dimension.Cation-selective exhaustive injection(CSEI) preconcentration method was used to introduce more analytes into the capillary.Furthermore,pH junction and sweeping dual concentration strategies were employed to avoid sample zone diffusion at the interface.The resulting electrophoregram was quite different from that of either CZE or MEKC separation.Up to(0.5-1.2) ×104 fold improvements in sensitivity were obtained relative to the conventional electrokinetic injection method.The proposed method was successfully applied to the determination of drugs in human urine.

  10. Determination of Amino Acids in Panax notoginseng by Microwave Hydrolysis and Derivatization Coupled with Capillary Zone Electrophoresis Detection

    Institute of Scientific and Technical Information of China (English)

    LI Xiao-tian; ZHAO Ya-jing; JIANG Cheng-fei; ZHANG Han-qi; YU Ai-min

    2013-01-01

    The microwave hydrolysis and derivatization coupled with capillary electrophoresis detection were developed for the separation and determination of the amino acids in Panax notoginseng.The experimental conditions for the microwave hydrolysis and derivatization were examined and optimized.Several parameters of capillary electrophoresis,such as pH value of background electrolyte,borate concentration and applied voltage were optimized.Under the selected conditions,11 amino acids were completely separated.The real sample was analyzed and the results were satisfactory.Compared with that of conventional heat hydrolysis and derivatization,the analytical time of this method was significantly shortened.

  11. Capillary electrophoresis of heparin and other glycosaminoglycans using a polyamine running electrolyte

    Energy Technology Data Exchange (ETDEWEB)

    Loegel, Thomas N.; Trombley, John D.; Taylor, Richard T. [Department of Chemistry and Biochemistry, Miami University, Oxford, OH 45056 (United States); Danielson, Neil D., E-mail: danielnd@muohio.edu [Department of Chemistry and Biochemistry, Miami University, Oxford, OH 45056 (United States)

    2012-11-13

    Highlights: Black-Right-Pointing-Pointer Ethylenediamine is likely acting as an ion-pairing agent. Black-Right-Pointing-Pointer Oversulfated chondroitin sulfate is last peak instead of first peak. Black-Right-Pointing-Pointer There is about a factor of five improved detectability with a 12.5 min analysis time. Black-Right-Pointing-Pointer Use of a 50 {mu}m ID capillary is possible. - Abstract: This study involves the use of polyamines as potential resolving agents for the capillary electrophoresis (CE) of glycosaminoglycans (GAGs), specifically heparin, dermatan sulfate, chondroitin sulfate, over-sulfated chondroitin sulfate (OSCS), and hyaluronan. All of the compounds can be separated from each other with the exception of chondroitin sulfate and hyaluronan. Using optimization software, the final run conditions are found to be 200 mM ethylenediamine and 45.5 mM phosphate as the electrolyte with -14 V applied across a 50 {mu}m ID Multiplication-Sign 24.5 cm fused silica capillary at 15 Degree-Sign C. The ion migration order, with OSCS as the last instead of the first peak, is in contrast to previous reports using either a high molarity TRIS or lithium phosphate run buffer with narrower bore capillaries. Total analysis time is 12. 5 min and the relative standard deviation of the heparin migration time is about 2.5% (n = 5). The interaction mechanism between selected polyamines and heparin is explored using conductivity measurements in addition to CE experiments to show that an ion-pairing mechanism is likely.

  12. High-Throughput Genetic Analysis and Combinatorial Chiral Separations Based on Capillary Electrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Wenwan Zhong

    2003-08-05

    Capillary electrophoresis (CE) offers many advantages over conventional analytical methods, such as speed, simplicity, high resolution, low cost, and small sample consumption, especially for the separation of enantiomers. However, chiral method developments still can be time consuming and tedious. They designed a comprehensive enantioseparation protocol employing neutral and sulfated cyclodextrins as chiral selectors for common basic, neutral, and acidic compounds with a 96-capillary array system. By using only four judiciously chosen separation buffers, successful enantioseparations were achieved for 49 out of 54 test compounds spanning a large variety of pKs and structures. Therefore, unknown compounds can be screened in this manner to identify optimal enantioselective conditions in just one rn. In addition to superior separation efficiency for small molecules, CE is also the most powerful technique for DNA separations. Using the same multiplexed capillary system with UV absorption detection, the sequence of a short DNA template can be acquired without any dye-labels. Two internal standards were utilized to adjust the migration time variations among capillaries, so that the four electropherograms for the A, T, C, G Sanger reactions can be aligned and base calling can be completed with a high level of confidence. the CE separation of DNA can be applied to study differential gene expression as well. Combined with pattern recognition techniques, small variations among electropherograms obtained by the separation of cDNA fragments produced from the total RNA samples of different human tissues can be revealed. These variations reflect the differences in total RNA expression among tissues. Thus, this Ce-based approach can serve as an alternative to the DNA array techniques in gene expression analysis.

  13. Recent advances in enhancing the sensitivity of electrophoresis and electrochromatography in capillaries and microchips (2012-2014).

    Science.gov (United States)

    Breadmore, Michael C; Tubaon, Ria Marni; Shallan, Aliaa I; Phung, Sui Ching; Abdul Keyon, Aemi S; Gstoettenmayr, Daniel; Prapatpong, Pornpan; Alhusban, Ala A; Ranjbar, Leila; See, Hong Heng; Dawod, Mohamed; Quirino, Joselito P

    2015-01-01

    One of the most cited limitations of capillary (and microchip) electrophoresis is the poor sensitivity. This review continues to update this series of biannual reviews, first published in Electrophoresis in 2007, on developments in the field of on-line/in-line concentration methods, covering the period July 2012-July 2014. It includes developments in the field of stacking, covering all methods from field-amplified sample stacking and large-volume sample stacking, through to ITP, dynamic pH junction, and sweeping. Attention is also given to on-line or in-line extraction methods that have been used for electrophoresis.

  14. Screening determination of pharmaceutical pollutants in different water matrices using dual-channel capillary electrophoresis coupled with contactless conductivity detection.

    Science.gov (United States)

    Le, Minh Duc; Duong, Hong Anh; Nguyen, Manh Huy; Sáiz, Jorge; Pham, Hung Viet; Mai, Thanh Duc

    2016-11-01

    In this study, the employment of purpose-made dual-channel compact capillary electrophoresis (CE) instrument with capacitively coupled contactless conductivity detection (C(4)D) as a simple and inexpensive solution for screening determination of various pharmaceutical pollutants frequently occurring in surface water and hospital wastewater in Hanoi, Vietnam is reported. Five negatively charged pharmaceutically active compounds, namely ibuprofen, diclofenac, bezafibrate, ketoprofen and mefenamic acid were determined using the first channel whereas three positively charged ones, namely diphenhydramine, metoprolol and atenolol were determined with the second channel of the CE-C(4)D instrument. Two different background electrolytes (BGEs) were used in these two CE channels independently. The best detection limits achieved were in the range of 0.2-0.8mg/L without sample pre-concentration. Enrichment factors up to 200 were obtainable with the inclusion of a solid phase extraction step. Good agreement between results obtained from CE-C(4)D and those with the standard confirmation method (HPLC-DAD) was achieved, with correlation coefficients higher than 0.98. PMID:27591645

  15. Extraction of rutin from buckwheat (Fagopyrum esculentumMoench) seeds and determination by capillary electrophoresis.

    Science.gov (United States)

    Kreft, S; Knapp, M; Kreft, I

    1999-11-01

    The content of the flavonoid rutin was determined in different milling fractions of buckwheat seeds and in buckwheat stems, leaves, and flowers. The extraction was performed by using a solvent containing 60% of ethanol and 5% of ammonia in water. The extracts were analyzed by capillary electrophoresis (running buffer of 50 mM borate (pH 9.3), 100 mM sodium dodecyl sulfate; determination at 380 nm). In bran fractions the concentration of rutin was 131-476 ppm, and in flour fractions 19-168 ppm. On average, about 300, 1000, and 46000 ppm of rutin were found in leaves, stems, and flowers, respectively. The results indicate that buckwheat could be an important nutritional source of flavonoids, especially in countries with a low mean daily flavonoid intake.

  16. Correlation between Molecular Structures and Relative Electrophoretic Mobility in Capillary Electrophoresis: Alkylpyridines

    Institute of Scientific and Technical Information of China (English)

    YAO, Xiao-Jun; FAN, Bo-Tao; DOUCET, J. P.; PANAYE, A.; LIU, Man-Cang; ZHANG, Rui-Sheng; HU, Zhi-De

    2003-01-01

    The quantitative relationship between relative electrophoretic mobility in capillary electrophoresis for a series of 31 closely related alkylpyridines and their molecular structures was studied by using CODESSA. According to the t-test on the results, we found that the three most important descriptors affecting the mobility are the relative number of rings (NR), Min e-n attraction for a C-N bond (MEN) and average complementary information index (ACIC). With these structure descriptors a good three-parameter linear model was developed to correlate the mobility of these compounds with their structures. This model can not only correctly predict the migration behavior of these compounds, but also find the structural factors which are responsible for the migration behavior of these compounds,thus can help to explain the separation mechanism of these compounds. The method used in this work can also be extended to the mobility-structure relationship research of other compounds.

  17. Capillary electrophoresis as a tool for the characterization of pentosan nanoparticles.

    Science.gov (United States)

    Abdel-Haq, Hanin; Bossù, Elena

    2012-09-28

    Because capillary zone electrophoresis (CZE) showed higher resolution for highly charged large carbohydrates and complex structures when compared to other chromatographic separation methods, it was chosen for the characterization of nanoparticles (NPs) of pentosan polysulfate (PPS). Thus, using the CZE technique, we developed a reliable, sensitive and rapid protocol that allowed the detection and characterization of PPS NPs. This protocol was able to determine the profile of both the NPs and the species of PPS entrapped into them, and to quantify free and bound PPS showing high reproducibility, acceptable accuracy and a good degree of precision. Moreover, it allowed the evaluation of the size and charge of the NPs. This protocol might be suitable for the characterization of other kinds of NPs also.

  18. Protein A Detection Based on Quantum Dots-Antibody Bioprobe Using Fluorescence Coupled Capillary Electrophoresis

    Directory of Open Access Journals (Sweden)

    Lin Qiu

    2014-01-01

    Full Text Available In this report, fluorescence detection coupled capillary electrophoresis (CE-FL was used to detect Protein A. Antibody was first labeled with Cy5 and then mixed with quantum dots (QDs to form QDs-antibody bioprobe. Further, we observed fluorescence resonance energy transfer (FRET from QDs donor to Cy5 acceptor. The bioprobe was formed and brought QDs and Cy5 close enough to allow FRET to occur. After adding protein A, the FRET system was broken and caused the FRET signal to decrease. Thus, a new method for the determination of protein A was proposed based on the FRET signal changes. This study provides a new trail of thought for the detection of protein.

  19. Capillary Electrophoresis Single-Strand Conformational Polymorphisms as a Method to Differentiate Algal Species

    Directory of Open Access Journals (Sweden)

    Alice Jernigan

    2015-01-01

    Full Text Available Capillary electrophoresis single-strand conformational polymorphism (CE-SSCP was explored as a fast and inexpensive method to differentiate both prokaryotic (blue-green and eukaryotic (green and brown algae. A selection of two blue-green algae (Nostoc muscorum and Anabaena inaequalis, five green algae (Chlorella vulgaris, Oedogonium foveolatum, Mougeotia sp., Scenedesmus quadricauda, and Ulothrix fimbriata, and one brown algae (Ectocarpus sp. were examined and CE-SSCP electropherogram “fingerprints” were compared to each other for two variable regions of either the 16S or 18S rDNA gene. The electropherogram patterns were remarkably stable and consistent for each particular species. The patterns were unique to each species, although some common features were observed between the different types of algae. CE-SSCP could be a useful method for monitoring changes in an algae species over time as potential shifts in species occurred.

  20. Low-temperature microchip nonaqueous capillary electrophoresis of aliphatic primary amines: applications to Titan chemistry.

    Science.gov (United States)

    Cable, Morgan L; Stockton, Amanda M; Mora, Maria F; Willis, Peter A

    2013-01-15

    We demonstrate microchip nonaqueous capillary electrophoresis (μNACE) analysis of primary aliphatic amines (C1-C18) in ethanol down to -20 °C as a first step in adapting microfluidic protocols for in situ analysis on Titan. To our knowledge, this is the first report of a nonaqueous separation at -20 °C on-chip. Limits of detection (LODs) ranged from 1.0 nM to 2.6 nM, and we identified several primary amines ranging in length from C2 to C16 in Titan aerosol analogue (tholin) samples; new amines were also detected in a tholin sample exposed to oxygen and liquid water. This preliminary work validates the sensitivity and efficacy of microfluidic chemical analysis of complex organics with relevance to Titan aerosols and surface deposits. PMID:23214444

  1. Development of a capillary electrophoresis method for the simultaneous determination of cephalosporins

    Directory of Open Access Journals (Sweden)

    Hancu Gabriel

    2013-01-01

    Full Text Available A rapid and simple capillary electrophoresis method has been developed for the simultaneous determination of six extensively used cephalosporin antibiotics (cefaclor, cefadroxil, cefalexin, cefuroxim, ceftazidim, ceftriaxon. The determination of cephalosporins was performed at a pH 6.8, using a 25 mM phospate - 25 mM borate mixed buffer, + 25 kV voltage at a temperature of 25 °C. We achieved a baseline separation in approximately 10 minutes. The separation resolution was increased by addition of an anionic surfactant, 50 mM sodium dodecyl sulfate, to the buffer solution. The proposed separation was evaluated on the basis of detection and quantification limits, effective electrophoretic mobility and relative standard deviation for migration times and peak areas.

  2. Determination of dissociation constants of pharmacologically active xanthones by capillary zone electrophoresis with diode array detection.

    Science.gov (United States)

    Wu, Xiaomu; Gong, Suxuan; Bo, Tao; Liao, Yiping; Liu, Huwei

    2004-12-24

    In this article, the dissociation constants (pKa) of 10 pharmacologically active xanthones isolated from herbal medicine Securidaca inappendiculata were determined by capillary zone electrophoresis with diode array detection. The pKa values determined by the method based on the electrophoretic mobilities (calculated from migration times) have been proved by the method based on UV absorbance calculated from the online spectra corresponding peaks. No conspicuous difference was observed between the two methods with acceptable reproducibility. Two pKa values (pKa1 and pKa2) were found for four xanthones while generally the 10 compounds possess the pKa values ranging from 6.4 to 9.2. PMID:15641365

  3. Detection of biogenic amines in C57BL/6 mice brain by capillary electrophoresis electrokinetic supercharging.

    Science.gov (United States)

    Wang, Wei-feng; Ju, Fu-rong; Ran, Yan-li; Zhang, Hui-ge; Chen, Xing-guo

    2016-02-01

    Ischemic stroke is caused when blood flow to the brain is stopped and is a major cause of death and long term disability across the globe. Excessive release of neurotransmitters is triggered in the brain by ischemia that mediates neuronal damage and causes ischemic injury. In this study, a simple, sensitive, and on-line preconcentration capillary electrophoresis method based on electrokinetic supercharging (EKS) was developed for the determination of the biogenic amines including dopamine (DA), epinephrine (E), and norepinephrine (NE) in C57BL/6 mice brain. Under the optimized conditions, the analytes were concentrated and detected within 10 min. The detection limits for the analytes ranged from 0.42 to 0.57 ng mL(-1) for a mice brain matrix. With the proposed method, the analyses of three neurochemical amines in C57BL/6 mice brain tissue during cerebral ischemic/reperfusion had been performed successfully. PMID:26658278

  4. Analysis of ecstasy tablets using capillary electrophoresis with capacitively coupled contactless conductivity detection.

    Science.gov (United States)

    Porto, Suely K S S; Nogueira, Thiago; Blanes, Lucas; Doble, Philip; Sabino, Bruno D; do Lago, Claudimir L; Angnes, Lúcio

    2014-11-01

    A method for the identification of 3,4-methylenedioxymethamphetamine (MDMA) and meta-chlorophenylpiperazine (mCPP) was developed employing capillary electrophoresis (CE) with capacitively coupled contactless conductivity detection (C(4) D). Sample extraction, separation, and detection of "Ecstasy" tablets were performed in Ecstasy tablets seized in Rio de Janeiro, Brazil, were analyzed by CE-C(4) D and compared against routine gas chromatography-mass spectrometry (GC-MS). The CE method demonstrated sufficient selectivity to discriminate the two target drugs, MDMA and mCPP, from the other drugs present in seizures, namely amphepramone, fenproporex, caffeine, lidocaine, and cocaine. Separation was performed in <90 sec. The advantages of using C(4) D instead of traditional CE-UV methods for in-field analysis are also discussed.

  5. Recent Advances in the Determination of Pesticides in Environmental Samples by Capillary Electrophoresis

    Directory of Open Access Journals (Sweden)

    Po-Ling Chang

    2016-04-01

    Full Text Available Nowadays, owing to the increasing population and the attempts to satisfy its needs, pesticides are widely applied to control the quantity and quality of agricultural products. However, the presence of pesticide residues and their metabolites in environmental samples is hazardous to the health of humans and all other living organisms. Thus, monitoring these compounds is extremely important to ensure that only permitted levels of pesticide are consumed. To this end, fast, reliable, and environmentally friendly methods that can accurately analyze dilute, complex samples containing both parent substances and their metabolites are required. Focusing primarily on research published since 2010, this review summarizes the use of various sample pretreatment techniques to extract pesticides from various matrices, combined with on-line preconcentration strategies for sensitivity improvement, and subsequent capillary electrophoresis analysis.

  6. Analysis of Phenolic Compounds in Coke Plant Wastewater by Capillary Zone Electrophoresis with Inhibited Chemiluminescence Detection

    Institute of Scientific and Technical Information of China (English)

    Xiang Dong XU; Yong Gang HU; Ze Yu YANG

    2006-01-01

    A capillary electrophoresis(CE) with on-line inhibited chemiluminescence (CL) detection was firstly used for the simultaneous analysis of benzenediol isomers and phenol. It is based on the quenching effect of benzenediol isomers and phenol on the chemiluminescence reaction of luminol with potassium ferricyanide in sodium hydroxide medium. Under the optimum conditions, the four phenols were baseline separated and detected in less than 10 min.The detection limits (S/N=3) for hydroquinone, resorcinol, catechol and phenol were 2.9×10-8mol/L, 3.7×10-7 mol/L, 8.4×10-8 mol/L and 4.4×10-6 mol/L, respectively. Finally, the presented method has been successfully applied to real sample.

  7. Micro-solid phase extraction of ochratoxin A, and its determination in urine using capillary electrophoresis

    International Nuclear Information System (INIS)

    We describe a simple, environmentally friendly and selective technique for the determination of ochratoxin A (OTA) in urine. It involves (a) the use of a molecularly imprinted polymer as a sorbent in micro-solid-phase extraction in which the sorbent is contained in a propylene membrane envelope, and (b) separation and detection by capillary electrophoresis (CE). Under optimized conditions, response is linear in the range between 50 and 300 ng mL−1 (with a correlation coefficient of 0.9989), relative standard deviations range from 4 to 8 %, the detection limit for OTA in urine is 11.2 ng mL−1 (with a quantification limits of 32.5 ng mL−1) which is lower than those of previously reported methods for solid-phase extraction combined with CE. The recoveries of OTA from urine spiked at levels of 50, 150 and 300 ng mL−1 ranged from 93 to 97 %. (author)

  8. Capillary electrophoresis with electrochemiluminescence detection for the analysis of quinolone drugs and pharmacokinetics study

    Institute of Scientific and Technical Information of China (English)

    Yan Ming Liu; Jun Tao Cao; Hui Wang

    2008-01-01

    A novel method for the determination of two quinolone drugs norfloxacin (NOR) and levofloxacin (LVX) was described by capillary electrophoresis with electrochemiluminescence detection. The good relationship (r ≥ 0.9991) between peak area and concentration of analytes was established over two orders of magnitude. The limits of detection (LOD, S/N = 3) in standard solution are 4.8 × 10-7 mol/L for NOR and 6.4 × 10-7 mol/L for LVX, respectively. The limits of quantitation (LOQ, S/N = 10) in real human urine samples are 1.2 × 10-6 mol/L for NOR and 1.4 × 10-6 mol/L for LVX, respectively. The present method was successfully applied to the determination of NOR and LVX in human urine and the study of pharmacokinetics of NOR.

  9. Application of 2,3-Naphthalenediamine in Labeling Natural Carbohydrates for Capillary Electrophoresis

    Directory of Open Access Journals (Sweden)

    Jim-Min Fang

    2012-06-01

    Full Text Available Neutral and acidic monosaccharide components in Ganoderma lucidum polysaccharide are readily labeled with 2,3-naphthalenediamine, and the resulting saccharide-naphthimidazole (NAIM derivatives are quantified by capillary electrophoresis (CE in borate buffer. Using sulfated-α-cyclodextrin as the chiral selector, enantiomers of monosaccharide-NAIMs are resolved on CE in phosphate buffer, allowing a simultaneous determination of the absolute configuration and sugar composition in the mucilage polysaccharide of a medicinal herb Dendrobium huoshanense. Together with the specific enzymatic reactions of various glycoside hydrolases on the NAIM derivatives of glycans, the structures of natural glycans can be deduced from the digestion products identified by CE analysis. Though heparin dissachrides could be successfully derived with the NAIM-labeling method, the heparin derivatives with the same degree of sulfation could not be separated by CE.

  10. Electrostatic interaction mechanism on the separation of phenols by non-aqueous capillary electrophoresis

    Institute of Scientific and Technical Information of China (English)

    WEI WeiLi; YIN YongGuang; XIA ZhiNing; CHEN ZhiTao; LIU WeiQi

    2007-01-01

    The electrostatic interaction between additive and analyte is of great importance to non-aqueous capillary electrophoresis (NACE) separation. Three tetraalkylammonium bromides and acetonitrile were applied as additives and running solvent respectively. The effect of alkyl chain length and concentration of additive on electrostatic interactions was investigated by the separation of phenols. The separation ability was found to increase with decreasing alkyl chain length of the additive, and the resolution values were increased with increasing additive concentration. The separation was seriously deteriorated after a little amount of water was added in the running solution. Furthermore, the electrostatic interaction is strong under the conditions of low electron cloud density, weak steric hindrance and multi-interaction sites. Thus, the separation result can be predicted by theoretical analysis, which is helpful for the separation of other substances in NACE based on electrostatic interaction.

  11. A multiscale products technique for denoising of DNA capillary electrophoresis signals

    Science.gov (United States)

    Gao, Qingwei; Lu, Yixiang; Sun, Dong; Zhang, Dexiang

    2013-06-01

    Since noise degrades the accuracy and precision of DNA capillary electrophoresis (CE) analysis, signal denoising is thus important to facilitate the postprocessing of CE data. In this paper, a new denoising algorithm based on dyadic wavelet transform using multiscale products is applied for the removal of the noise in the DNA CE signal. The adjacent scale wavelet coefficients are first multiplied to amplify the significant features of the CE signal while diluting noise. Then, noise is suppressed by applying a multiscale threshold to the multiscale products instead of directly to the wavelet coefficients. Finally, the noise-free CE signal is recovered from the thresholded coefficients by using inverse dyadic wavelet transform. We compare the performance of the proposed algorithm with other denoising methods applied to the synthetic CE and real CE signals. Experimental results show that the new scheme achieves better removal of noise while preserving the shape of peaks corresponding to the analytes in the sample.

  12. Signal Detection of Multi-Channel Capillary Electrophoresis Chip Based on CCD

    Science.gov (United States)

    Lv, Hongfeng; Yan, Weiping; Yang, Xiaobo; Li, Jiechao; Zhu, Jieying

    2012-12-01

    A kind of multi-channel capillary electrophoresis (CE) chip signal detection system based on CCD was developed. The output signal of the CCD sensor was processed by a series of pre-processing circuits and ADC, and then it was collected by the Field Programmable Gate Array (FPGA) chip which communicated with a host computer. The core in FPGA was designed to control the signal flow of the CCD and transfer the data to PC based on a Nios II embedded soft-processor. The application of PC was used to store the data and demonstrate the curve. The measurement of the fluorescent signals for different concentration Rhodamine B dyes is presented and the comparison with other detection systems is also discussed.

  13. Automatic Combination of Microfluidic Nanoliter-Scale Droplet Array with High-Speed Capillary Electrophoresis

    Science.gov (United States)

    Li, Q.; Zhu, Y.; Zhang, N.-Q.; Fang, Q.

    2016-05-01

    In this paper, we developed a novel approach for interfacing a microfluidic two-dimensional droplet array to a high-speed capillary electrophoresis (HSCE) system. Picoliter-scale sample injection (ca. 200 pL) from a nanoliter-scale droplet array covered by nonvolatile oil was automatically achieved using the spontaneous injection mode, without the interference from the cover oil and the need of special droplet extraction interface as in previously reported systems. The system was applied in consecutive separations of 25 different samples of amino acids with a whole separation time less than 15 min, as well as on-line monitoring of in-droplet derivatizing reaction of amino acids by fluorescein isothiocyanate (FITC) over 3 hours. High separation speed (up to 100 samples per hour) and high separation efficiency (up to 9.22 × 105 N/m) were achieved.

  14. Hydrogel plug for independent sample and buffer handling in continuous microchip capillary electrophoresis

    Science.gov (United States)

    Puchberger-Enengl, Dietmar; Bipoun, Mireille; Smolka, Martin; Krutzler, Christian; Keplinger, Franz; Vellekoop, Michael J.

    2013-05-01

    In microchip capillary electrophoresis most frequently electrokinetic sample injection is utilized, which does not allow pressure driven sample handling and is sensitive for pressure drops due to different reservoir levels. For efficient field tests a multitude of samples have to be processed with the least amount of external equipment. We present the use of a hydrogel plug to separate the sample from clean buffer to enable independent sample change and buffer refreshment. In-situ polymerization of the gel does away with complex membrane fabrication techniques. The sample is electrokinetically injected through the gel and subsequently separated by a voltage between the second gel inlet and the buffer outlet. By blocking of disturbing flows by the gel barrier a well-defined ion plug is obtained. After each experiment, the sample and the separation channel can be flushed independently, allowing for a continuous operation mode in order to process multiple samples.

  15. Capillary electrophoresis-electrochemistry microfluidic system for the determination of organic peroxides

    Science.gov (United States)

    Wang, Joseph; Escarpa, Alberto; Pumera, Martin; Feldman, Jason; Svehla, D. (Principal Investigator)

    2002-01-01

    A microfluidic analytical system for the separation and detection of organic peroxides, based on a microchip capillary electrophoresis device with an integrated amperometric detector, was developed. The new microsystem relies on the reductive detection of both organic acid peroxides and hydroperoxides at -700 mV (vs. Ag wire/AgCl). Factors influencing the separation and detection processes were examined and optimized. The integrated microsystem offers rapid measurements (within 130 s) of these organic-peroxide compounds, down to micromolar levels. A highly stable response for repetitive injections (RSD 0.35-3.12%; n = 12) reflects the negligible electrode passivation. Such a "lab-on-a-chip" device should be attractive for on-site analysis of organic peroxides, as desired for environmental screening and industrial monitoring.

  16. Detection of Elevated Signaling Amino Acids in Human Diabetic Vitreous by Rapid Capillary Electrophoresis

    Directory of Open Access Journals (Sweden)

    Miao-Jen Lu

    2007-01-01

    Full Text Available Elevated glutamate is implicated in the pathology of PDR. The ability to rapidly assess the glutamate and amino acid content of vitreous provides a more complete picture of the chemical changes occurring at the diabetic retina and may lead to a better understanding of the pathology of PDR. Vitreous humor was collected following vitrectomies of patients with PDR and control conditions of macular hole or epiretinal membrane. A capillary electrophoresis method was developed to quantify glutamate and arginine. The analysis is relatively fast (<6 minutes and utilizes a poly(ethyleneoxide and sodium dodecylsulfate run buffer. Both amino acid levels show significant increases in PDR patients versus controls and are comparable to other reports. The levels of vitreal glutamate vary inversely with the degree of observed hemorrhage. The results demonstrate a rapid method for assessment of a number of amino acids to characterize the chemical changes at the diabetic retina to better understand tissue changes and potentially identify new treatments.

  17. Investigation of interaction between the drug and cell membrane by capillary electrophoresis

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    By introducing cell membrane into electrophoretic buffer as pseudo-stationary phase,a novel capillary electrophoresis method was established to explore the interaction between drugs and cell membrane,where the interaction between citalopram and rabbit red blood cell membrane was used as an example. A series of concentrations of cell membrane were suspended into the running buffer by peak-shift method. The binding constant of citalopram to rabbit red blood cell membrane of 0.977 g-1·L was obtained after treatment of Scatchard plot. This method could provide not only a new way for the investigation on the interactions between drugs and cell membrane,but also a new approach for high throughput screening of the drug membrane permeability,biological activity,and evaluating drugs in vivo.

  18. Functional integration of PCR amplification and capillary electrophoresis in a microfabricated DNA analysis device.

    Science.gov (United States)

    Woolley, A T; Hadley, D; Landre, P; deMello, A J; Mathies, R A; Northrup, M A

    1996-12-01

    Microfabricated silicon PCR reactors and glass capillary electrophoresis (CE) chips have been successfully coupled to form an integrated DNA analysis system. This construct combines the rapid thermal cycling capabilities of microfabricated PCR devices (10 degrees C/s heating, 2.5 degrees C/s cooling) with the high-speed (Real-time monitoring of PCR target amplification in these integrated PCR-CE devices is also feasible. Amplification of the beta-globin target as a function of cycle number was directly monitored for two different reactions starting with 4 x 10(7) and 4 x 10(5) copies of DNA template. This work establishes the feasibility of performing high-speed DNA analyses in microfabricated integrated fluidic systems. PMID:8946790

  19. Determination of Magnolol and Honokiol by Non-aqueous Capillary Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Two active principles in traditional Chinese medicine Magnolia officinalis, magnolol and honokiol, were successfully separated by means of nonaqueous capillary electrophoresis. The effect of the composition of a nonaqueous buffer on column efficiency and resolution, and the effect of acid additives on peak shapes were researched. The separation was conducted with a running buffer in a mixture of methanol/acetonitrile/formamide (volume ratio: 1: 2: 2), in which the concentrations of Tris, acetic acid, and water were 60 mmol/L, 0. 04 mmol/L and 5% (volume fration),respectively, and the pH * (apperent pH) of the running buffer was 8. 96. Magnolol and honokiol were separated on baseline within 20 min. The relative standard deviation of the analytes' concentrations in the sample is 1.32% for magnolol and 1.60% for honokiol, and the recoveries of the spiked sample are 98.4% for magnolol and 98.0% for honokiol, respectively.

  20. Typing of 49 autosomal SNPs by single base extension and capillary electrophoresis for forensic genetic testing.

    Science.gov (United States)

    Børsting, Claus; Tomas, Carmen; Morling, Niels

    2012-01-01

    We describe a method for simultaneous amplification of 49 autosomal single nucleotide polymorphisms (SNPs) by multiplex PCR and detection of the SNP alleles by single base extension (SBE) and capillary electrophoresis. All the SNPs may be amplified from only 100 pg of genomic DNA and the length of the amplicons range from 65 to 115 bp. The high sensitivity and the short amplicon sizes make the assay very suitable for typing of degraded DNA samples, and the low mutation rate of SNPs makes the assay very useful for relationship testing. Combined, these advantages make the assay well suited for disaster victim identifications, where the DNA from the victims may be highly degraded and the victims are identified via investigation of their relatives. The assay was validated according to the ISO 17025 standard and used for routine case work in our laboratory. PMID:22139655

  1. Analysis of Glutamic Acid in Cerebrospinal Fluid by Capillary Electrophoresis with High Frequency Conductivity Detection

    Institute of Scientific and Technical Information of China (English)

    Hai Yun ZHAI; Jun Mei WANG; Xiao Li YAO; Xue Cai TAN; Pei Xiang CAI; Zuan Guang CHEN

    2005-01-01

    A rapid method to determine glutamic acid (Glu) in cerebrospinal fluid (CSF) by capillary electrophoresis with high frequency conductivity detection (contactless conductivity detection) was described. The CSF sample was pretreated with silver cation resin to remove high concentration of Cl- ions in CSF. The separation was achieved in the buffer solution of 10 mmol/L Tris and 8 mmol/L boric acid at the separation voltage of 20.0 kV. Glu showed linear response in the range of 5.0×10-6 to 6.0×10-3 mol/L, the limit of detection was 1.0×10-6 mol/L. The method was used for analysis Glu in CSF satisfactorily with a recovery of 97.8-98.8%.

  2. Determination of metoprolol in rabbit blood using capillary electrophoresis with laser-induced fluorescence detection

    Institute of Scientific and Technical Information of China (English)

    Yu Yun Chen; Wei Ping Yang; Zhu Jun Zhang

    2011-01-01

    This work described a sensitive method for determination of metoprolol in rabbit plasma. The method involved purification by ultrafiltration, derivatization with fluorescein isothiocyanate, determination by capillary electrophoresis (CE) coupled with laser-induced fluorescence (LIF) detector. Other components in plasma including a variety of amino acids and proteins did not interfere with the determination of metoprolol in experimental condition. The assay had a wide range (2.0-500 ng/mL) of linearity and a detection limit of 0.8 ng/mL. The intra-and inter-day precisions were satisfactory with relative standard deviation (RSD) less than 10.0% and accuracy within 10.0%. This method was successfully applied to pharmacokinetic study of metoprolol in rabbit blood. (c) 2010 Yu Yun Chen. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

  3. Advances in chiral separations of small peptides by capillary electrophoresis and chromatography.

    Science.gov (United States)

    Ali, Imran; Al-Othman, Zeid A; Al-Warthan, Abdulrahman; Asnin, Leonid; Chudinov, Alexander

    2014-09-01

    Many chemical and biological processes are controlled by the stereochemistry of small polypeptides (di-, tri-, tetra-, penta-, hexapeptides, etc). The biological importance of peptide stereoisomers is of great value. Therefore, the chiral resolution of peptides is an important issue in biological and medicinal sciences and drug industries. The chiral resolutions of peptide racemates have been discussed with the use of capillary electrophoresis and chromatographic techniques. The various chiral selectors used were polysaccharides, cyclodextrins, Pirkle types, macrocyclic antibiotics, crown ethers, imprinted polymers, etc. The stereochemistry of dipeptides is also discussed. Besides, efforts are made to explain the chiral recognition mechanisms, which will be helpful in understanding existing and developing new stereoselective analyses. Future perspectives of enantiomeric resolution are also predicted. Finally, the review concludes with the demand of enantiomeric resolution of all naturally occurring and synthetic peptides. PMID:25044566

  4. Reliable low-cost capillary electrophoresis device for drug quality control and counterfeit medicines.

    Science.gov (United States)

    Marini, R D; Rozet, E; Montes, M L A; Rohrbasser, C; Roht, S; Rhème, D; Bonnabry, P; Schappler, J; Veuthey, J-L; Hubert, Ph; Rudaz, S

    2010-12-15

    The proportion of counterfeit medicines is dramatically increasing these last few years. According to numerous official sources, in some pharmaceutical wholesalers in African countries, the proportion has reached 80%. Unfortunately, this situation is far to be improved due to lack of suitable analytical equipment allowing rapid actions of the Regulatory Agencies based on scientific consideration, at affordable cost and all over the drug supply chain. For that purpose, a network group considered that mater by building a low-cost original capillary electrophoresis (CE) equipment equipped with a new deep UV detector based on LED technology. The generic conditions for analysis were investigated: capillary zone electrophoresis (CZE) performed at acidic pH for basic drug molecules (i.e., quinine, highly used as the last antimalarial rampart), basic pH for compounds such as furosemide (a common diuretic drug) and at neutral pH for a well known antibiotic combination, trimethoprim/sulfamethoxazol. To evaluate the ability of the CE equipment for quantification, a full validation and a method comparison study were carried out for the CZE method dedicated to quinine determination. The validation involved the use of accuracy profile and total error concept to monitor the adequacy of the results obtained by the new prototype. The method comparison was based on the Bland and Altman approach by comparing results obtained by the low-cost CE and a conventional set-up. Subsequent validation studies were realized with neutral and acidic drug molecules, each focusing on a single concentration level calibration curve in order to maintain as low as possible the expenses due to reagents and thus the cost of analysis, as important advantages of CE for drug quality control. PMID:20719445

  5. Powder-blasting technology as an alternative tool for microfabrication of capillary electrophoresis chips with integrated conductivity sensors

    NARCIS (Netherlands)

    Schlautmann, Stefan; Wensink, Henk; Schasfoort, Richard; Elwenspoek, Miko; Berg, van den Albert

    2001-01-01

    The fabrication and characterization of a microfluidic device for capillary electrophoresis applications is presented. The device consists of a glass chip which contains a single separation channel as well as an integrated conductivity detection cell. In contrast to most microfluidic glass devices t

  6. Electrochemical Detection of Alkaline Phosphatase in BALB/c Mouse Fetal Liver Stromal Cells with Capillary Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    Xue Mei SUN; Dong LI; Zeng Liang BAI; Wen Rui JIN

    2004-01-01

    A method for determination of alkaline phosphatase (ALP) in BALB/c mouse fetal liver stromal cells has been described based on the catalytic reaction. After the cell extract is incubated with the substrate disodium phenyl phosphate, the reaction product phenol generated by ALP is determined by capillary electrophoresis with electrochemical detection.

  7. Determination of the enantiomeric purity of (-) terbutaline by capillary electrophoresis using cyclodextrins as chiral selectors in a polyethylene glycol gel

    NARCIS (Netherlands)

    de Boer, T; Ensing, K

    1998-01-01

    A method was developed for determination of the enantiomeric purity of the therapeutic-pharmacological active (-)-enantiomer of terbutaline using cyclodextrins as a chiral selector dissolved in a removable liquid polyethylene glycol gel by use of capillary electrophoresis. The effect of temperature,

  8. Tuning of the selectivity in capillary electrophoresis by cyclodextrins illustrated by the separation of some structurally related phenothiazine

    NARCIS (Netherlands)

    de Boer, T; Bijma, R; Ensing, K

    1998-01-01

    Cyclodextrins were used to affect the selectivity of the capillary electrophoresis system in the separation of 10 widely used phenothiazines. It was shown that the addition of cyclodextrins substantially improved the selectivity. The effect of temperature and cyclodextrin concentration was studied o

  9. DNA Sequencing by Capillary Electrophoresis Using Quasi-inter penetrating Network Formed by Polyacrylamide and Poly(N-hydroxymethylacrylamide)

    Institute of Scientific and Technical Information of China (English)

    Wen Long ZHANG; Yan Mei WANG

    2006-01-01

    Quasi-interpenetrating network formed by polyacrylamide and poly (N-hydroxymethylacrylamide) was designed, synthesized, and tested for DNA sequencing by capillary electrophoresis. The performance of quasi-IPN on DNA sequencing was determined by the acrylamide to N-hydroxymethylacrylamide molar ratio and sequencing temperature.

  10. Determination of Roxithromycin Tablets by Capillary Electrophoresis Employing Non-aqueous Media with Square-wave Amperometric Detection

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    A new method of determination for roxithromycin tablets by non-aqueous capillary electrophoresis (NACE) with square-wave amperometric detection was carried out. Several parameters affecting the NACE-AD determination were studied. The data was modified by spline wavelet least square (SWLS). The method is simple, rapid and highly reliable for routine analysis.

  11. Fluorescence- and capillary electrophoresis (CE)-based SSR DNA fingerprinting and a molecular identity database for the Louisiana sugarcane industry

    Science.gov (United States)

    A database of Louisiana sugarcane molecular identity has been constructed and is being updated annually using FAM or HEX or NED fluorescence- and capillary electrophoresis (CE)-based microsatellite (SSR) fingerprinting information. The fingerprints are PCR-amplified from leaf DNA samples of current ...

  12. A chip-type thin-layer electrochemical cell coupled with capillary electrophoresis for online separation of electrode reaction products

    Energy Technology Data Exchange (ETDEWEB)

    He, Jian-Bo, E-mail: jbhe@hfut.edu.cn; Cui, Ting; Zhang, Wen-Wen; Deng, Ning

    2013-07-05

    Graphical abstract: -- Highlights: •A new coupling of thin-layer electrolysis with capillary electrophoresis (CE). •Rapid electrolysis, direct sampling followed by online CE separation. •At least 13 products of quercetin oxidation were separated. •Thermodynamic and kinetic parameters were determined from CE peak areas. -- Abstract: A coupling technique of thin-layer electrolysis with high-performance capillary electrophoresis/UV–vis technique(EC/HPCE/UV–vis) is developed for online separation and determination of electrode reaction products. A chip-type thin-layer electrolytic (CTE) cell was designed and fabricated, which contains a capillary channel and a background electrolyte reservoir, allowing rapid electrolysis, direct sampling and online electrophoretic separation. This chip-type setup was characterized based on an electrophoresis expression of Nernst equation that was applied to the redox equilibrium of o-tolidine at different potentials. The utility of the method was demonstrated by separating and determining the electro-oxidation products of quercetin in different pH media. Two main products were always found in the studied time, potential and pH ranges. The variety of products increased not only with increasing potential but also with increasing pH value, and in total, at least 13 products were observed in the electropherograms. This work illustrates a novel example of capillary electrophoresis used online with thin-layer electrolysis to separate and detect electrode reaction products.

  13. A capillary zone electrophoresis method to detect conformers and dimers of antithrombin in therapeutic preparations.

    Science.gov (United States)

    Marie, Anne-Lise; Tran, Nguyet Thuy; Saller, François; Abdou, Youmna Mohamed; Zeau, Pascal; Plantier, Jean-Luc; Urbain, Rémi; Borgel, Delphine; Taverna, Myriam

    2016-07-01

    Antithrombin (AT) is a human plasma glycoprotein that possesses anticoagulant and anti-inflammatory properties. However, the native (active) form of AT is unstable and undergoes conformational changes, leading to latent, cleaved, and heterodimeric forms. The presence of these alternative forms mostly inactive can highly impact the quality and therapeutic activity of pharmaceutical AT preparations. We developed a capillary zone electrophoresis method, based on a neutral polyethylene oxide-coated capillary and a buffer close to physiological conditions, enabling the separation of more than eight forms of AT. Several peaks were identified as native, latent, and heterodimeric forms. The CZE method was reproducible with intraday relative standard deviations less than 0.5 and 2% for migration times and peak areas, respectively. The method was applied to the comparison of AT preparations produced by five competitive pharmaceutical companies, and statistical tests were performed. Important differences in the proportion of each form were highlighted. In particular, one AT preparation was shown to contain a high quantity of heterodimer, and two preparations contained high quantities of latent form. In addition, one AT preparation exhibited additional forms, not yet identified. PMID:26989842

  14. Development of a fast capillary electrophoresis method to determine inorganic cations in biodiesel samples.

    Science.gov (United States)

    Piovezan, Marcel; Costa, Ana Carolina O; Jager, Alessandra Vincenzi; de Oliveira, Marcone Augusto Leal; Micke, Gustavo Amadeu

    2010-07-19

    The aim of this study was to develop a fast capillary electrophoresis method for the determination of inorganic cations (Na(+), K(+), Ca(2+), Mg(2+)) in biodiesel samples, using barium (Ba(2+)) as the internal standard. The running electrolyte was optimized through effective mobility curves in order to select the co-ion and Peakmaster software was used to determine electromigration dispersion and buffer capacity. The optimum background electrolyte was composed of 10 mmol L(-1) imidazole and 40 mmol L(-1) of acetic acid. Separation was conducted in a fused-silica capillary (32 cm total length and 23.5 cm effective length, 50 microm I.D.), with indirect UV detection at 214 nm. The migration time was only 36 s. In order to obtain the optimized conditions for extraction, a fractional factorial experimental design was used. The variables investigated were biodiesel mass, pH, extractant volume, agitation and sonication time. The optimum conditions were: biodiesel mass of 200 mg, extractant volume of 200 microL and agitation of 20 min. The method is characterized by good linearity in the concentration range of 0.5-20 mg kg(-1) (r>0.999), limit of detection was equal to 0.3 mg kg(-1), inter-day precision was equal to 1.88% and recovery in the range of 88.0-120%. The developed method was successfully applied to the determination of cations in biodiesel samples. PMID:20599036

  15. Analyses of Phytohormones in Coconut (Cocos Nucifera L. Water Using Capillary Electrophoresis-Tandem Mass Spectrometry

    Directory of Open Access Journals (Sweden)

    Swee Ngin Tan

    2014-12-01

    Full Text Available Capillary electrophoresis (CE coupled with mass spectrometry (MS or tandem mass spectrometry (MS/MS is reported as an alternative and potentially useful method for the simultaneous analysis of various classes of phytohormones with diversified structures, including indole-3-acetic acid (IAA, indole-3-butyric acid (IBA, abscisic acid (ABA, gibberellic acid (GA, zeatin (Z, N6-benzyladenine (BA, α-naphthaleneacetic acid (NAA and 2,4-dichlorophenoxyacetic acid (2,4-D. The key to the CE-MS/MS analysis was based on electroosmotic flow reversal using a cationic polymer-coated capillary. Under optimum conditions, a baseline separation of eight phytohormones was accomplished within 30 min using 60 mM ammonium formate/formic acid buffer of pH 3.8 with −20 kV as the separation voltage. The accessibility of MS/MS together with the characterization by migration properties obtained by CE allows for the development of CE-MS/MS as an emerging potential method for the analysis of different classes of phytohormones in a single run. The utility of the CE-MS/MS method was demonstrated by the comprehensive screening of phytohormones in coconut (Cocos nucifera L. water after pre-concentration and purification through solid-phase extraction (SPE cartridge. IAA, ABA, GA and Z were detected and quantified in the purified coconut water extract sample.

  16. [The determination of glucose, sucrose and fructose by the method of capillary electrophoresis].

    Science.gov (United States)

    Yakuba, Yu F; Markovsky, M G

    2015-01-01

    The possibilities of different regimes of micellar capillary electrophoresis using negative polarity and alkaline electrolyte for determination of glucose, sucrose, fructose in extracts of vegetative organs of plants and products of fruits and grapes processing have been studied. A comparative evaluation of the limits of detection of glucose, sucrose, fructose for developed electrolytes have been performed, the advantages and disadvantages of techniques have been discussed. It is recommended to use an aqueous electrolyte containing 0.5% potassium sorbate, 0.62% cetyltrimethylammonium bromide, and 0.02% potassium hydroxide. The analyzed components were detected at 254 nm. The sample was dosed hydrodynamically (30 mbar, 5 sec). Negative voltage 16 kV is recommended, current--54 ± 4 µA, capillary thermostating at 24 °C is applied, the analysis time--15 min. The detection limits for fructose and glucose is 0.03 g/dm3 to 0.07 g of sucrose/dm3. Linearity is stored for each component to 5.0 g/dm 3 inclusive. Electrophoretic mobility of carbohydrates was (10(-4) sm2V(-1)sec(-1)): fructose--3.12, glucose--3.03, sucrose--2.74. Approximate time of release: glucose--13 min, sucrose--13.5 min, fructose--12.5 min. The developed options for mass concentration determining of mono- and disaccharides provide complete separation of the components. Anions, glycerol, ethylene glycol, propylene glycol and butylene isomers do not affect the analysis results. PMID:26402948

  17. Influence of Analyte Concentration on Stability Constant Values Determined by Capillary Electrophoresis.

    Science.gov (United States)

    Sursyakova, Viktoria V; Burmakina, Galina V; Rubaylo, Anatoly I

    2016-08-01

    The influence of analyte concentration when compared with the concentration of a charged ligand in background electrolyte (BGE) on the measured values of electrophoretic mobilities and stability constants (association, binding or formation constants) is studied using capillary electrophoresis (CE) and a dynamic mathematical simulator of CE. The study is performed using labile complexes (with fast kinetics) of iron (III) and 5-sulfosalicylate ions (ISC) as an example. It is shown that because the ligand concentration in the analyte zone is not equal to that in BGE, considerable changes in the migration times and electrophoretic mobilities are observed, resulting in systematic errors in the stability constant values. Of crucial significance is the slope of the dependence of the electrophoretic mobility decrease on the ligand equilibrium concentration. Without prior information on this dependence to accurately evaluate the stability constants for similar systems, the total ligand concentration must be at least >50-100 times higher than the total concentration of analyte. Experimental ISC peak fronting and the difference between the direction of the experimental pH dependence of the electrophoretic mobility decrease and the mathematical simulation allow assuming the presence of capillary wall interaction. PMID:27090731

  18. Determination of cellular carbohydrates in peanut fungal pathogens and baker's yeast by capillary electrophoresis and electrochromatography.

    Science.gov (United States)

    Zhang, M; Melouk, H A; Chenault, K; El Rassi, Z

    2001-11-01

    In this work, the quantitation of cellular carbohydrates, namely chitin and glucan, in peanut fungal pathogens and baker's yeast was carried out by capillary electrophoresis (CE) and capillary electrochromatography (CEC). The chitin and glucan of the fungi were hydrolyzed by the enzymes chitinase and glucanase, respectively, to their corresponding sugar monomers N-acetylglucosamine (GlcNAc) and glucose (Glc). These two monosaccharides were then tagged with 6-aminoquinoline (6-AQ) to allow their separation and detection in CE and CEC. The 6-AQ derivatives of GlcNAc and Glc formed the basis for the determination by CE and CEC of chitin and glucan in peanut fungi and baker's yeast. Several parameters affecting the separation of the 6-AQ derivatives of GlcNAc and Glc, including the separation voltage and the composition of the running electrolyte, were investigated. Under the optimized separation conditions, the contents of cellular carbohydrates including N-acetylglucosamine, chitin, glucose, and glucan in some fungi, such as Sclerotinia minor, Sclerotium rolfsii, and baker's yeast, were successfully determined. The method described here allowed the assessment of genetic differences in Sclerotium rolfsii isolates from various locations. PMID:11714314

  19. Simultaneous determination of melamine and 5-hydroxymethylfurfural in milk by capillary electrophoresis with diode array detection.

    Science.gov (United States)

    Chen, Zhijun; Yan, Xiaomei

    2009-10-14

    This article describes the development of a simple analytical approach for the simultaneous determination of melamine and 5-hydroxymethylfurfural (HMF) in milk samples using capillary electrophoresis (CE) with diode array detection (DAD) for the first time. Ultraviolet absorption at wavelengths of 214 and 280 nm was applied for the detection of melamine and HMF, respectively. Milk samples were extracted with 1% trichloroacetic acid using a high-speed blender and ultrasonication. After centrifugation and filtration, the extract was analyzed by CE-DAD directly. Micellar electrokinetic capillary chromatography was employed as the separation mode by adding sodium dodecyl sulfate (SDS) to the electrolyte. Under optimal separation conditions, melamine, HMF, and interferents were well resolved. The linear dynamic ranges were 0.05-100 microg/mL for melamine (R(2) = 0.9996) and 0.1-100 microg/mL for HMF (R(2) = 0.9997). The assay detection limits were 0.047 microg/mL and 0.067 microg/mL for melamine and HMF, respectively. Satisfactory results were obtained for the assay recovery rate and repeatability. The proposed method was successfully applied for the analysis of melamine and HMF in real milk samples, and the results of melamine were comparable to those obtained using HPLC-UV reference method. PMID:19761188

  20. Capillary electrophoresis separation of neutral organic compounds, pharmaceutical drugs, proteins and peptides, enantiomers, and anions

    Energy Technology Data Exchange (ETDEWEB)

    Ding, W.L.

    1999-02-12

    Addition of a novel anionic surfactant, namely lauryl polyoxyethylene sulfate, to an aqueous-acetonitrile electrolyte makes it possible to separate nonionic organic compounds by capillary electrophoresis. Separation is based on differences in the association between analytes and the surfactant. Highly hydrophobic compounds such as polyaromatic hydrocarbons are well separated by this new surfactant. Migration times of analytes can be readily changed over an unusually large range by varying the additive concentration and the proportion of acetonitrile in the electrolyte. Several examples are given, including the separation of four methylbenz[a]anthracene isomers and the separation of normal and deuterated acetophenone. The effect of adding this new surfactant to the acidic electrolyte was also investigated. Incorporation of cetyltrimethylammonium bromide in the electrolyte is shown to dynamically coat the capillary and reverse electroosmotic flow. Chiral recognition mechanism is studied using novel synthetic surfactants as chiral selectors, which are made from amino acids reacting with alkyl chloroformates. A satisfactory separation of both inorganic and organic anions is obtained using electrolyte solutions as high as 5 M sodium chloride using direct photometric detection. The effect of various salts on electrophoretic and electroosmotic mobility is further discussed. Several examples are given under high-salt conditions.

  1. Multi-Site N-glycan mapping study 1: Capillary electrophoresis – laser induced fluorescence

    Science.gov (United States)

    Szekrényes, Ákos; Park, SungAe Suhr; Santos, Marcia; Lew, Clarence; Jones, Aled; Haxo, Ted; Kimzey, Michael; Pourkaveh, Shiva; Szabó, Zoltán; Sosic, Zoran; Feng, Peng; Váradi, Csaba; de l'Escaille, François; Falmagne, Jean-Bernard; Sejwal, Preeti; Niedringhaus, Thomas; Michels, David; Freckleton, Gordon; Hamm, Melissa; Manuilov, Anastasiya; Schwartz, Melissa; Luo, Jiann-Kae; van Dyck, Jonathan; Leung, Pui-King; Olajos, Marcell; Gu, Yingmei; Gao, Kai; Wang, Wenbo; Wegstein, Jo; Tep, Samnang; Guttman, András

    2016-01-01

    An international team that included 20 independent laboratories from biopharmaceutical companies, universities, analytical contract laboratories and national authorities in the United States, Europe and Asia was formed to evaluate the reproducibility of sample preparation and analysis of N-glycans using capillary electrophoresis of 8-aminopyrene-1,3,6-trisulfonic acid (APTS)-labeled glycans with laser induced fluorescence (CE-LIF) detection (16 sites) and ultra high-performance liquid chromatography (UHPLC, 12 sites; results to be reported in a subsequent publication). All participants used the same lot of chemicals, samples, reagents, and columns/capillaries to run their assays. Migration time, peak area and peak area percent values were determined for all peaks with >0.1% peak area. Our results demonstrated low variability and high reproducibility, both, within any given site as well across all sites, which indicates that a standard N-glycan analysis platform appropriate for general use (clone selection, process development, lot release, etc.) within the industry can be established. PMID:26466659

  2. Capillary electrophoresis as a novel technique for screening natural flavonoids as kinase inhibitors.

    Science.gov (United States)

    Nehmé, Reine; Nehmé, Hala; Roux, Grégory; Destandau, Emilie; Claude, Bérengère; Morin, Philippe

    2013-11-29

    Capillary electrophoresis (CE) was used for the first time to evaluate the inhibition activity of aglycone flavonoids (such as quercetin and isorhamnetin) and some of their glycosylated derivatives toward human kinases. The cyclin-dependant kinase 5 (CDK5/p25) and the glycogen synthase kinase 3β (GSK3β) were chosen since they are very promising biological targets for developing treatments against neurodegenerative diseases and cancer. In a previous work, we developed an in-capillary kinase CE assay where the capillary was used as an enzymatic nanoreactor in which the kinase, its substrate, adenosine 5'-triphosphate (ATP) and its potential inhibitor were mixed by using transverse diffusion of laminar flow profiles (TDLFP). The product adenosine 5'-diphosphate (ADP) was then detected at 254nm and quantified. In this work, this assay was improved to reduce, for the first time, the dilution effect commonly observed with the TDLFP approach. Under the new conditions established herein, IC50 values for quercetin, kaempferol and flavopiridol were successfully obtained and were in the same order of magnitude of those reported in the literature using the conventional assay using radioactive (33)P-ATP. It was shown that aglycone flavonoids have an inhibition activity more important than their glycosylated derivatives. CE was also proved to be very efficient for evaluating inhibition activity of complex samples such as crude extracts of sea buckthorn (SBT) berries obtained by solvent-free microwave extraction (SFME). This novel approach to combine SFME technique to a CE-based enzymatic assay is very interesting for evaluating the biological activity of natural material in a fast, simple, economic (no use of neither fluorescent nor radiometric labels) and green (no organic solvents) manner. PMID:24148298

  3. Rapid enantioseparation of amlodipine by highly sulfated cyclodextrins using short-end injection capillary electrophoresis

    Directory of Open Access Journals (Sweden)

    M Zandkarimi

    2009-12-01

    Full Text Available "n  "n Background and the purpose of the study:The use of highly sulfated cyclodextrins (HS-CDs as chiral selectors in capillary electrophoresis (CE has been examined for rapid and reproducible enantioseparation of the model drug amlodipine, a calcium channel blocker. "nMaterials and Methods: Fused silica capillaries with an inner diameter of 50 μm, and a total length of 45.5 cm (8.5 cm to the detector were used. Capillaries were rinsed with polyethylene oxide (PEO once daily. A systematic method development approach was conducted by modifying selected parameters such as the type and concentration of the chiral selector, the buffer pH and concentration of the background electrolyte. "nResults: Baseline separation was achieved at low (i.e. 0.05%w/v concentrations of HS-αCD, but migration time and peak area repeatability were more than 4% and 25% of the relative standard deviation (RSD, respectively. At higher concentrations (>0.3% of HS-αCD, amlodipine was transported to the anode by the carrier ability of HS-αCD. In carrier mode, the migration order of enantiomers was reversed, the migration time was reduced and the peak area repeatability of analysis was improved. The optimum electrophoretic conditions for the stereoselective analysis of amlodipine were obtained in carrier mode with 25 mM sodium phosphate buffer containing 1.25% w/v of HS-αCD at pH 2.5 with an applied voltage of +15 kV. Under these conditions migration time was less than 3 min and within-day migration time and peak area repeatability, were less than 0.4% and 2.1% RSD, respectively. Conclusions: Rapid enantioseparation was achieved with minimum variation in quantitative analysis. These optimized conditions are appropriate for the enantioselective analysis of amlodipine.

  4. Protein analysis by membrane preconcentration-capillary electrophoresis: systematic evaluation of parameters affecting preconcentration and separation.

    Science.gov (United States)

    Rohde, E; Tomlinson, A J; Johnson, D H; Naylor, S

    1998-08-25

    Fast and efficient analysis of proteins in physiological fluids is of great interest to researchers and clinicians alike. Capillary electrophoresis (CE) has proven to be a potentially valuable tool for the separation of proteins in specimens. However, a generally acknowledged drawback of this technique is the limited sample volumes which can be loaded onto the CE capillary which results in a poor concentration limit of detection. In addition, matrix components in samples may also interfere with separation and detection of analytes. Membrane preconcentration-CE (mPC-CE) has proved to be effective in overcoming these problems. In this report, we describe the systematic evaluation of parameters affecting on-line preconcentration/clean-up and separation of protein mixtures by mPC-CE. Method development was carried out with a standard mixture of proteins (lysozyme, myoglobin, carbonic anhydrase, and human serum albumin). First, using MALDI-TOF-MS, membrane materials with cation-exchange (R-SO3H) or hydrophobic (C2, C8, C18, SDB) characteristics were evaluated for their potential to retain proteins in mPC cartridges. Hydrophobic membranes were found most suitable for this application. Next, all mPC-CE analysis of protein samples were performed in polybrene coated capillaries and parameters affecting sample loading, washing and elution, such as the composition and volume of the elution solvent were investigated. Furthermore, to achieve optimal mPC-CE performance for the separation of protein mixtures parameters affecting postelution focusing and electrophoresis, including the composition of the background electrolyte and a trailing stacking buffer were varied. Optimal conditions for mPC-CE analysis of proteins using a C2 impregnated membrane preconcentration (mPC) cartridge were achieved with a background electrolyte of 5% acetic acid and 2 mM ammonium acetate, 60 nl of 80% acetonitrile in H2O as an elution solvent, and 60 nl of 0.5% ammonium hydroxide as a trailing

  5. Determination of arsenic species in Solanum Lyratum Thunb using capillary electrophoresis with inductively coupled plasma mass spectrometry.

    Science.gov (United States)

    Shuai, Pei-Yu; Yang, Xiao-Jun; Qiu, Zong-Qing; Wu, Xiao-Hui; Zhu, Xi; Pokhrel, Ganga Raj; Fu, Yu-Ying; Ye, Hui-Min; Lin, Wen-Xiong; Yang, Gui-Di

    2016-08-01

    A simple and highly efficient interface to couple capillary electrophoresis with inductively coupled plasma mass spectrometry by a microflow polyfluoroalkoxy nebulizer and a quadruple ion deflector was developed in this study. By using this interface, six arsenic species, including arsenite, arsenate, monomethylarsonic acid, dimethylarsinic acid, arsenobetaine, and arsenocholine, were baseline-separated and determined in a single run within 11 min under the optimized separation conditions. The instrumental detection limit was in the range of 0.02-0.06 ng/mL for the six arsenic compounds. Repeatability expressed as the relative standard deviation (n = 5) of both migration time and peak area were better than 2.5 and 4.3% for six arsenic compounds. The proposed method, combined with a closed-vessel microwave-assisted extraction procedure, was successfully applied for the determination of arsenic species in the Solanum Lyratum Thunb samples from Anhui province in China with the relative standard deviations (n = 5) ≤4%, method detection limits of 0.2-0.6 ng As/g and a recovery of 98-104%. The experimental results showed that arsenobetaine was the main speciation of arsenic in the Solanum Lyratum Thunb samples from different provinces in China, with a concentration of 0.42-1.30 μg/g. PMID:27378629

  6. Improvement on Simultaneous Determination of Cr(III) and Cr(VI) by Capillary Electrophoresis and Chemiluminescence Detection

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    A sensitive method for the simultaneous determination of Cr(III) and Cr(VI) using in-capillary reaction capillary electrophoresis separation and chemiluminescence detection was developed. The procedures were designed as follows: The sample, hydrochloric acid and sodium hydrogen sulfite solution segments were injected sequentially into the capillary. The reaction of Cr(VI) reduced to Cr(III) by HSO3- occurred inside the capillary after applying the running voltage. According to the migration time difference of both Cr(III) ions moving towards to the cathode (detection end), they could be separated and determined. The limits of detection for chromium(III) and chromium(VI) (S/N = 3) were 6.0(10-13 mol/L (12 zmol) and 1.9(10-11 mol/L (380 zmol), respectively.

  7. Automation and integration of polymerase chain reaction with capillary electrophoresis for high throughput genotyping and disease diagnosis

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, N.

    1999-02-12

    Genotyping is to detect specific loci in the human genome. These loci provide important information for forensic testing, construction of genetic linkage maps, gene related disease diagnosis and pharmacogenetic research. Genotyping is becoming more and more popular after these loci can be easily amplified by polymerase chain reaction (PCR). Capillary electrophoresis has its unique advantages for DNA analysis due to its fast heat dissipation and ease of automation. Four projects are described in which genotyping is performed by capillary electrophoresis emphasizing different aspects. First, the author demonstrates a principle to determine the genotype based on capillary electrophoresis system. VNTR polymorphism in the human D1S80 locus was studied. Second, the separation of four short tandem repeat (STR) loci vWF, THO1, TPOX and CSF1PO (CTTv) by using poly(ethylene oxide) (PEO) was studied in achieving high resolution and preventing rehybridization of the DNA fragments. Separation under denaturing, non-denaturing conditions and at elevated temperature was discussed. Third, a 250 {micro}m i.d., 365 {micro}m o.d. fused silica capillary was used as the microreactor for PCR. Fourth, direct PCR from blood was studied to simplify the sample preparation for genotyping to minimum.

  8. CEval: All-in-one software for data processing and statistical evaluations in affinity capillary electrophoresis.

    Science.gov (United States)

    Dubský, Pavel; Ördögová, Magda; Malý, Michal; Riesová, Martina

    2016-05-01

    We introduce CEval software (downloadable for free at echmet.natur.cuni.cz) that was developed for quicker and easier electrophoregram evaluation and further data processing in (affinity) capillary electrophoresis. This software allows for automatic peak detection and evaluation of common peak parameters, such as its migration time, area, width etc. Additionally, the software includes a nonlinear regression engine that performs peak fitting with the Haarhoff-van der Linde (HVL) function, including automated initial guess of the HVL function parameters. HVL is a fundamental peak-shape function in electrophoresis, based on which the correct effective mobility of the analyte represented by the peak is evaluated. Effective mobilities of an analyte at various concentrations of a selector can be further stored and plotted in an affinity CE mode. Consequently, the mobility of the free analyte, μA, mobility of the analyte-selector complex, μAS, and the apparent complexation constant, K('), are first guessed automatically from the linearized data plots and subsequently estimated by the means of nonlinear regression. An option that allows two complexation dependencies to be fitted at once is especially convenient for enantioseparations. Statistical processing of these data is also included, which allowed us to: i) express the 95% confidence intervals for the μA, μAS and K(') least-squares estimates, ii) do hypothesis testing on the estimated parameters for the first time. We demonstrate the benefits of the CEval software by inspecting complexation of tryptophan methyl ester with two cyclodextrins, neutral heptakis(2,6-di-O-methyl)-β-CD and charged heptakis(6-O-sulfo)-β-CD.

  9. Functionalization and characterization of persistent luminescence nanoparticles by dynamic light scattering, laser Doppler and capillary electrophoresis.

    Science.gov (United States)

    Ramírez-García, Gonzalo; d'Orlyé, Fanny; Gutiérrez-Granados, Silvia; Martínez-Alfaro, Minerva; Mignet, Nathalie; Richard, Cyrille; Varenne, Anne

    2015-12-01

    Zinc gallate nanoparticles doped with chromium (III) (ZnGa1.995O4:Cr0.005) are innovative persistent luminescence materials with particular optical properties allowing their use for in vivo imaging. They can be excited in the tissue transparency window by visible photons and emit light for hours after the end of the excitation. This allows to observe the probe without any time constraints and without autofluorescence signals produced by biological tissues. Modification of the surface of these nanoparticles is essential to be colloidally stable not only for cell targeting applications but also for proper distribution in living organisms. The use of different methods for controlling and characterizing the functionalization process is imperative to better understand the subsequent interactions with biological elements. This work explores for the first time the characterization and optimization of a classic functionalization sequence, starting with hydroxyl groups (ZGO-OH) at the nanoparticle surface, followed by an aminosilane-functionalization intermediate stage (ZGO-NH2) before PEGylation (ZGO-PEG). Dynamic light scattering and laser doppler electrophoresis were used in combination with capillary electrophoresis to characterize the nanoparticle functionalization processes and control their colloidal and chemical stability. The hydrodynamic diameter, zeta potential, electrophoretic mobility, stability over time and aggregation state of persistent luminescence nanoparticles under physiological-based solution conditions have been studied for each functional state. Additionally, a new protocol to improve ZGO-NH2 stability based on a thermal treatment to complete covalent binding of (3-aminopropyl) triethoxysilane onto the particle surface has been optimized. This thorough control increases our knowledge on these nanoparticles for subsequent toxicological studies and ultimately medical application.

  10. Analysis of Trinitrophenylated Adenosine and Inosine by Capillary Electrophoresis and γ-Cyclodextrin-Enhanced Fluorescence Detection.

    Science.gov (United States)

    Stephen, Terilyn K L; Guillemette, Katherine L; Green, Thomas K

    2016-08-01

    Monitoring molecules such as adenosine (Ado) and inosine (Ino) in the central nervous system has enabled the field of neuroscience to correlate molecular concentrations dynamics to neurological function, behavior, and disease. In vivo sampling techniques are commonly used to monitor these dynamics; however, many techniques are limited by the sensitivity and sample volume requirements of currently available detection methods. Here, we present a novel capillary electrophoresis-laser-induced fluorescence detection (CE-LIF) method that analyzes Ado and Ino by derivatization with 2,4,6-trinitrobenzenesulfonic acid to form fluorescent trinitrophenylated complexes of Ado (TNP-Ado) and Ino (TNP-Ino). These complexes exhibit ∼25-fold fluorescence enhancement upon the formation of inclusion complexes with γ-cyclodextrin (γ-CD). Association constants were determined as 4600 M(-1) for Ado and 1000 M(-1) for Ino by CE-LIF. The structure of the TNP-Ado:γ-CD complex was determined by 2D nuclear magnetic resonance (NMR) spectroscopy. Optimal trinitrophenylation reaction conditions and CE-LIF parameters were determined and resulted in the limit of detection of 1.6 μM for Ado and 4 μM for Ino. Ado and Ino were simultaneously quantified in homogenized rat forebrain samples to illustrate application of the technique. Simulated biological samples, desalted by ultrafiltration in the presence γ-CD, were concentrated on-capillary by large-volume sample stacking (LVSS) to achieve detection limits of 32 and 38 nM for TNP-Ado and TNP-Ino, respectively. PMID:27314490

  11. Portable, Battery Operated Capillary Electrophoresis with Optical Isomer Resolution Integrated with Ionization Source for Mass Spectrometry

    Science.gov (United States)

    Moini, Mehdi; Rollman, Christopher M.

    2016-03-01

    We introduce a battery operated capillary electrophoresis electrospray ionization (CE/ESI) source for mass spectrometry with optical isomer separation capability. The source fits in front of low or high resolution mass spectrometers similar to a nanospray source with about the same weight and size. The source has two high voltage power supplies (±25 kV HVPS) capable of operating in forward or reverse polarity modes and powered by a 12 V rechargeable lithium ion battery with operation time of ~10 h. In ultrafast CE mode, in which short narrow capillaries (≤15 μm i.d., 15-25 cm long) and field gradients ≥1000 V/cm are used, peak widths at the base are <1 s wide. Under these conditions, the source provides high resolution separation, including optical isomer resolution in ~1 min. Using a low resolution mass spectrometer (LTQ Velos) with a scan time of 0.07 s/scan, baseline separation of amino acids and their optical isomers were achieved in ~1 min. Moreover, bovine serum albumin (BSA) was analyzed in ~1 min with 56% coverage using the data-dependent MS/MS. Using a high resolution mass spectrometer (Thermo Orbitrap Elite) with 15,000 resolution, the fastest scan time achieved was 0.15 s, which was adequate for CE-MS analysis when optical isomer separation is not required or when the optical isomers were well separated. Figures of merit including a detection limit of 2 fmol and linear dynamic range of two orders of magnitude were achieved for amino acids.

  12. Physico-chemical characterization of liposomes and drug substance-liposome interactions in pharmaceutics using capillary electrophoresis and electrokinetic chromatography

    DEFF Research Database (Denmark)

    Franzen, Ulrik; Østergaard, Jesper

    2012-01-01

    of liposome drug delivery systems, e.g., for the investigation of encapsulation efficiency and drug leakage. The well-known characteristics of capillary electrophoresis, i.e., low sample volume requirement, high separation efficiency in aqueous media without a stationary phase, minimal sample preparation......Liposomes are self-assembled phospholipid vesicles and have numerous research and therapeutic applications. In the pharmaceutical and biomedical sciences liposomes find use as models of biological membranes, partitioning medium and as drug carriers. The present review addresses the use of capillary...

  13. Precision improvement for the analysis of flavonoids in selected Thai plants by capillary zone electrophoresis.

    Science.gov (United States)

    Suntornsuk, Leena; Anurukvorakun, Oraphan

    2005-02-01

    A capillary zone electrophoresis (CZE) method for the analyses of kaempferol in Centella asiatica and Rosa hybrids and rutin in Chromolaena odorata was developed. The optimization was performed on analyses of flavonoids (e.g., rutin, kaempferol, quercetin, myricetin, and apigenin) and organic carboxylic acids (e.g., ethacrynic acid and xanthene-9-carboxylic acid) by investigation of the effects of types and amounts of organic modifiers, background electrolyte concentrations, temperature, and voltage. Baseline separation (R(s) = 2.83) of the compounds was achieved within 10 min in 20 mM NaH2PO4 - Na2HPO4 (pH 8.0) containing 10% v/v ACN and 6% v/v MeOH using a voltage of 25 kV, a temperature of 30 degrees C, and a detection wavelength set at 220 nm. The application of the corrected migration time (t(c)), using ethacrynic acid as the single marker, was efficient to improve the precision of flavonoid identification (% relative standard deviation (RSD) = 0.65%). The method linearity was excellent (r2 > 0.999) over 50-150 microg/mL. Precision (%RSD 96% and %RSDs odorata was 0.088 g/100 g (%RSD = 0.06%). PMID:15690438

  14. Real-time monitoring of peptide grafting onto chitosan films using capillary electrophoresis.

    Science.gov (United States)

    Taylor, Danielle L; Thevarajah, Joel J; Narayan, Diksha K; Murphy, Patricia; Mangala, Melissa M; Lim, Seakcheng; Wuhrer, Richard; Lefay, Catherine; O'Connor, Michael D; Gaborieau, Marianne; Castignolles, Patrice

    2015-03-01

    Chitosan, being antimicrobial and biocompatible, is attractive as a cell growth substrate. To improve cell attachment, arginine-glycine-aspartic acid-serine (RGDS) peptides were covalently grafted to chitosan films, through the widely used coupling agents 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC-HCl) and N-hydroxysuccinimide (NHS), via the carboxylic acid function of the RGDS molecule. The grafting reaction was monitored, for the first time, in real time using free-solution capillary electrophoresis (CE). This enabled fast separation and determination of the peptide and all other reactants in one separation with no sample preparation. Covalent RGDS peptide grafting onto the chitosan film surface was demonstrated using solid-state NMR of swollen films. CE indicated that oligomers of RGDS, not simply RGDS, were grafted on the film, with a likely hyperbranched structure. To assess the functional properties of the grafted films, cell growth was compared on control and peptide-grafted chitosan films. Light microscopy and polymerase chain reaction (PCR) analysis demonstrated greatly improved cell attachment to RGDS-grafted chitosan films. PMID:25680633

  15. Capillary zone electrophoresis to study humic fraction in forest soils and its relationship with radionuclide mobility

    International Nuclear Information System (INIS)

    In order to determine the humic acid content in soil samples, a methodology has been developed using capillary zone electrophoresis (CZE), which is a technique with high separation efficiency and resolution. Working conditions of CZE separation were optimized and procedure was established. This methodology was applied to study the influence of organic matter matter (mainly humic acids) on the retention of radionuclides. The design mentioned above provided us with a great number of samples. For each sampling, samples were taken from the different horizons in several points. The fraction of radionuclide incorporated to each horizon was measured. The percentage of radionuclide associated with humic acids was estimated by means of single extractions using suitable reagents. The dark-coloured supernatant was separated from the residue by centrifugation and radionuclide content was quantified by gamma spectrometry. Humic content in these extracts was determined by acidifying with HCl to pH = 1 and allowing it to stand at room temperature for 16 hours. In this way, humic acid was obtained in a coagulated form, dissolved in NaOH solution and subsequently estimated using the CZE technique. After these measurements it is possible to test whether there is any correlation between humic acid and radionuclide contents in each sample and to obtain conclusions about the role of organic matter in radionuclide retention. 1 fig

  16. Capillary Electrophoresis-Mass Spectrometry for the Analysis of Heparin Oligosaccharides and Low Molecular Weight Heparin.

    Science.gov (United States)

    Sun, Xiaojun; Lin, Lei; Liu, Xinyue; Zhang, Fuming; Chi, Lianli; Xia, Qiangwei; Linhardt, Robert J

    2016-02-01

    Heparins, highly sulfated, linear polysaccharides also known as glycosaminoglycans, are among the most challenging biopolymers to analyze. Hyphenated techniques in conjunction with mass spectrometry (MS) offer rapid analysis of complex glycosaminoglycan mixtures, providing detailed structural and quantitative data. Previous analytical approaches have often relied on liquid chromatography (LC)-MS, and some have limitations including long separation times, low resolution of oligosaccharide mixtures, incompatibility of eluents, and often require oligosaccharide derivatization. This study examines the analysis of glycosaminoglycan oligosaccharides using a novel electrokinetic pump-based capillary electrophoresis (CE)-MS interface. CE separation and electrospray were optimized using a volatile ammonium bicarbonate electrolyte and a methanol-formic acid sheath fluid. The online analyses of highly sulfated heparin oligosaccharides, ranging from disaccharides to low molecular weight heparins, were performed within a 10 min time frame, offering an opportunity for higher-throughput analysis. Disaccharide compositional analysis as well as top-down analysis of low molecular weight heparin was demonstrated. Using normal polarity CE separation and positive-ion electrospray ionization MS, excellent run-to-run reproducibility (relative standard deviation of 3.6-5.1% for peak area and 0.2-0.4% for peak migration time) and sensitivity (limit of quantification of 2.0-5.9 ng/mL and limit of detection of 0.6-1.8 ng/mL) could be achieved.

  17. Poly(ethylene glycol)-functionalized polymeric microchips for capillary electrophoresis.

    Science.gov (United States)

    Sun, Xuefei; Li, Dan; Lee, Milton L

    2009-08-01

    Recently, we reported the synthesis, fabrication, and preliminary evaluation of poly(ethylene glycol) (PEG)-functionalized polymeric microchips that are inherently resistant to protein adsorption without surface modification in capillary electrophoresis (CE). In this study, we investigated the impact of cross-linker purity and addition of methyl methacrylate (MMA) as a comonomer on CE performance. Impure poly(ethylene glycol) diacrylate (PEGDA) induced electroosmotic flow (EOF) and increased the separation time, while the addition of MMA decreased the separation efficiency to approximately 25% of that obtained using microchips fabricated without MMA. Resultant improved microchips were evaluated for the separation of fluorescent dyes, amino acids, peptides, and proteins. A CE efficiency of 4.2 x 10(4) plates for aspartic acid in a 3.5 cm long microchannel was obtained. Chiral separation of 10 different D,L-amino acid pairs was obtained with addition of a chiral selector (i.e., beta-cyclodextrin) in the running buffer. Selectivity (alpha) and resolution (R(s)) for D,L-leucine were 1.16 and 1.64, respectively. Good reproducibility was an added advantage of these PEG-functionalized microchips.

  18. Rapid simultaneous determination of organic acids, free amino acids, and lactose in cheese by capillary electrophoresis.

    Science.gov (United States)

    Izco, J M; Tormo, M; Jiménez-Flores, R

    2002-09-01

    A capillary electrophoresis (CE) method for the simultaneous separation of 11 metabolically important organic acids (oxalic, formic, citric, succinic, orotic, uric, acetic, pyruvic, propionic, lactic, and butyric), 10 amino acids (Asp, Glu, Tyr, Gly, Ala, Ser, Leu, Phe, Lys, and Trp), and lactose has been optimized, validated, and tested in dairy products. Repeatability and linearity were calculated for each compound, with detection limit values as low as 0.2 x 10(-2) mM for citric acid and Gly. The method was applied to analyze yogurt and different varieties of commercial cheeses. This method yielded specific CE patterns for different varieties of cheese. Also, it has been shown to be sensitive enough to measure small changes in composition of some of those compounds in fresh cheese stored under accelerated ripening conditions for 2 d at 32 degrees C (e.g., from 1728.3 +/- 45.0 to 1166.7 +/- 4.5 mg/100 g of DM in the case of lactose, or from 23.5 +/- 0.6 to 76.8 +/- 16.7 mg/100 g of DM in the case of acetic acid).

  19. Metabolomics, peptidomics and proteomics applications of capillary electrophoresis-mass spectrometry in Foodomics: A review

    Energy Technology Data Exchange (ETDEWEB)

    Ibáñez, Clara; Simó, Carolina; García-Cañas, Virginia; Cifuentes, Alejandro, E-mail: a.cifuentes@csic.es; Castro-Puyana, María

    2013-11-13

    Graphical abstract: -- Highlights: •Foodomics allows studying food and nutrition through the application of advanced omics approaches. •CE-MS plays a crucial role as analytical platform to carry out omics studies. •CE-MS applications for food metabolomics, proteomics and peptidomics are presented. -- Abstract: In the current post-genomic era, Foodomics has been defined as a discipline that studies food and nutrition through the application of advanced omics approaches. Foodomics involves the use of genomics, transcriptomics, epigenetics, proteomics, peptidomics, and/or metabolomics to investigate food quality, safety, traceability and bioactivity. In this context, capillary electrophoresis-mass spectrometry (CE-MS) has been applied mainly in food proteomics, peptidomics and metabolomics. The aim of this review work is to present an overview of the most recent developments and applications of CE-MS as analytical platform for Foodomics, covering the relevant works published from 2008 to 2012. The review provides also information about the integration of several omics approaches in the new Foodomics field.

  20. Digitally synchronized LCD projector for multi-color fluorescence excitation in parallel capillary electrophoresis detection.

    Science.gov (United States)

    Lin, Shi-Wei; Chang, Chih-Hang; Wu, Dai-Yang; Lin, Che-Hsin

    2010-10-15

    A simple method is proposed for modulating the excitation light used for multi-color fluorescence detection in a single capillary electrophoresis (CE) channel. In the proposed approach, a low-cost commercial liquid crystal device (LCD) projector with digitally-modulated LCD switches is used to provide the illumination light source and the fluorescence emitted from the CE chip is synchronously detected using an ultraviolet-visible-near infrared (UV-vis-NIR) spectrometer. The modulated light source enables the detection of multiple fluorescence signals within a single CE channel without the need of mechanically switching optical components. In order to enhance the sensing performance of the proposed system, two short-pass filters and one band-pass filter are inserted into the LCD projector to modify the wavelength spectra for fluorescence excitation. With this simple approach, the signal-to-noise (SN) ratio of the fluorescence detection signals is greatly improved by a factor of approximately 22 when detecting Atto647N fluorescent dye. The feasibility of the proposed multi-color CE detection approach is demonstrated by detecting two different samples including a mixed sample comprising FITC, Rhodamine B and Atto647N fluorescent dyes and a bio-sample composed of two ssDNAs labeled with FITC and Cy3, respectively. Results confirm that the digitally-modulated excitation system proposed in this study has significant potential for the parallel analysis of fluorescently-labeled bio-samples using a multi-color detection scheme.

  1. [Recent advances in the application of high performance capillary electrophoresis for food safety].

    Science.gov (United States)

    Dong, Yalei; Chen, Xiaojiao; Hu, Jing; Chen, Xingguo

    2012-11-01

    In recent years, food safety incidents become a serious social problem. Foods are usually complex mixtures consisting of a large diversity of molecules. Analysis of foods is a topic that demands the development of rapid, robust, efficient, sensitive and cost-effective analytical methodologies. Therefore, new techniques for food safety purpose are required by analytical chemists. Capillary electrophoresis (CE) is a popular separation technique that possesses fast and efficient performances in an automated way with minimum consumption of sample and reagents. Nowadays, CE represents a desired strategy for the determination of many compounds or molecules in various kinds of food. In this paper, the review intends to provide the recent innovative developments reported in food safety analysis using CE methods for a full overview. As a fundamental review, it focuses on the introduction and detection of several common hazardous materials existing in food such as non-food additives, pesticide residues, veterinary drug residues, heavy metal ion contaminants, toxins, biphenol A and phthalates in packaging materials and so on. Furthermore, this review prospects the main development direction of CE in this field for the future. A total of 63 papers published during the period of Jan 2009 - Jun 2012 are included in the present review. PMID:23451513

  2. Determination of active ingredients of Rhododendron dauricum L. by capillary electrophoresis with electrochemical detection.

    Science.gov (United States)

    Cao, Yuhua; Lou, Changgang; Fang, Yuzhi; Ye, Jiannong

    2002-01-11

    High-performance capillary electrophoresis with electrochemical detection was employed to analyse active ingredients of Rhododendron dauricum L., an important crude herb frequently used in Chinese medicines. Farrerol, quercetin, syringic acid, vanillic acid, 4-hydroxybenzoic acid, protocatechuic acid are major important active ingredients. Operated in a wall-jet configuration, a 300-microm diameter carbon-disk electrode was used as the working electrode, which exhibits a good response at +950 mV (vs. saturated calomel electrodes) for six analytes. Under the optimum conditions, the analytes were baseline separated within 16 min in a borax buffer (pH 8.7). Notably, excellent linearity was obtained over two orders of magnitude with detection limits (S/N=3) ranged from 9 x 10(-7) to 3.0 x 10(-6) M for all analytes. This method was successfully used in the analysis of Rhododendron dauricum L. with relatively simple extraction procedures, and the assay results were satisfactory. PMID:11820277

  3. Determination of phycobiliproteins by capillary electrophoresis with laser-induced fluorescence detection.

    Science.gov (United States)

    Viskari, P J; Kinkade, C S; Colyer, C L

    2001-07-01

    Phycobiliproteins are derived from the photosynthetic apparatus of cyanobacteria and eukaryotic algae. They are composed of a protein backbone to which linear tetrapyrrole chromophores are covalently bound. Furthermore, they are water-soluble highly fluorescent, and relatively stable at room temperature and neutral pH. For this reason, capillary electrophoresis-laser induced fluorescence (CE-LIF) seems the idea method for determination of these important proteins. The effects of buffer additives such as sodium dodecyl sulfate (SDS)and putrescine on the separation of the three major phycobiliprotein types, namely allophycocyanin, phycocyanin, and phycoerythrin, with excitation and emission maxima at 652/660, 615/647, and 565(494)/575 nm, respectively, are considered. Detection limits for these proteins by CE-LIF are some 60-500 times better than by absorbance detection. The development of a fast and sensitive CE-LIF assay such as this is of potential significance to our understand ing of chemical and biological oceanographic processes.

  4. Analysis of Trace Ingredients in Green Tea by Capillary Electrophoresis with Amperometric Detection

    Institute of Scientific and Technical Information of China (English)

    LI Ping; DONG Shu-Qing; WANG Qing-Jiang; FANG Yu-Zhi

    2008-01-01

    In this paper, four trace ingredients (rutin, gallic acid, quercetin, chlorogenic acid) in green tea were simultaneously determined by capillary electrophoresis coupled with amperometric detection (CE-AD). Effects of several important factors such as the pH and concentration of running buffer, separation voltage, injection time and detection potential were investigated to acquire the optimum conditions. Under the optimum conditions, the analytes could be separated within 20 min at a separation voltage of 18 kV in a 60 mmol/L borate buffer (pH 8.7). A 300 μmdiameter carbon disk electrode generated good responses at 950 mV (vs. SCE) for all analytes. The relationship between the peak currents and concentrations of the analytes was linear over about three orders of magnitude with demonstrated long-term stability and reproducibility with relative standard deviations less than 3% for both migration time and peak current (n=7), which could be successfully used for the determination of the analytes in green tea with satisfactory assay results.

  5. Immunochromatographic removal of albumin in erythropoietin biopharmaceutical formulations for its analysis by capillary electrophoresis.

    Science.gov (United States)

    Lara-Quintanar, Pilar; Lacunza, Izaskun; Sanz, Jesus; Diez-Masa, Jose Carlos; de Frutos, Mercedes

    2007-06-15

    Human serum albumin (HSA) is added to some pharmaceutical preparations as an excipient. This is the case for some of the commercial preparations of recombinant erythropoietin (rEPO). Differences in the number of the sialic acid moieties in the different rEPO glycoforms confer to these forms different net charges and different bioactivity. Knowledge of the isoforms present in each pharmaceutical product is then of interest. Differences in net charge of the rEPO forms make possible their separation by electrophoretical methods. However it has been observed in our laboratory that the amount of HSA usually present in these drug formulations interferes or even precludes separation of rEPO bands by capillary zone electrophoresis (CZE). In this work, an immunochromatographic method to remove HSA from rEPO biopharmaceutical formulations and a procedure to concentrate the sample that is needed to be performed prior to the analysis by CZE are developed. A home-made computer program to compare the percentage of correct assignments of electrophoretical bands provided by different migration parameters is used to study the effect of HSA remaining in samples on the accuracy of assignment of rEPO bands. When there exists a residual concentration of HSA in the sample (studies and for the quality control laboratories of the manufacturers. PMID:16919660

  6. Determination of Four Active Ingredients in Vc Yinqiao Tablets by Capillary Zone Electrophoresis with Amperometric Detection

    Institute of Scientific and Technical Information of China (English)

    L(U),Jin; WANG,Qing-Jiang; CHENG,Xi; LIU,Hai-Yan; HE,Pin-Gang; FANG,Yu-Zhi

    2006-01-01

    A simple, reliable and reproducible method, based on capillary zone electrophoresis with amperometric detection (CZE-AD), has been developed for simultaneous determination of four active ingredients in Vc Yinqiao tablets including paracetamol, vitamin C, caffeic acid and chlorogenic acid. A carbon-disk electrode was used as working electrode and 0.95 V (versus SCE) was selected as detection potential. The optimal conditions of CZE experiment were 30 mmol·L-1 borate solution (pH 9.5) as running buffer, 14 kV as separation voltage and 8 s (14 kV) as electro-kinetic sampling time. Under the selected optimum conditions, paracetamol, vitamin C, caffeic acid and chlorogenic acid could be perfectly separated within 22 min, and their detection limits (S/N=3) ranged from 5 × 10-7 to 1×10-6 mol·L-1. This proposed method demonstrated good reproducibility with relative standard deviations of less than 3% for both migration time and peak current (n=7). The utility of this method was demonstrated by monitoring a kind of compound medicine named Vc Yinqiao tablets and the assay results were satisfactory.

  7. Determination of Active Ingredients of Hawthorn by Capillary Electrophoresis with Electrochemical Detection

    Institute of Scientific and Technical Information of China (English)

    TANG Zhu-Xing; ZENG Yi-Kun; ZHOU Yun; ZANG Shu-Liang; HE Pin-Gang; FANG Yu-Zhi

    2006-01-01

    A method based on capillary electrophoresis with electrochemical detection has been developed for the separation and determination of epicatechin, kaempferol, chlorogenic acid, 4-hydroxybenzoic acid, quercetin and protocatechuic acid in hawthorn for the first time. The effects of working electrode potential, pH and concentration of running buffer, separation voltage and injection time on CE-ED were investigated. Under the optimum conditions,the analytes could be separated in a 60 mmol·L-1 borate buffer (pH 8.7) within 21 min. A 300 μm diameter carbon disk electrode has a good response at +0.95 V (vs. SCE) for all analytes. The response was linear over three orders of magnitude with detection limits (S/N=3) ranging from 3×10-8 to2×10-7 g·mL-1 for the analytes. The method has been successfully applied to the analysis of real sample, with satisfactory results.

  8. Using capillary electrophoresis to study the chemical conditions within cracks in aluminum alloys.

    Science.gov (United States)

    Cooper, K R; Kelly, R G

    1999-07-30

    The environment-assisted cracking (EAC) susceptibility of some aluminum alloys used for airplane structural components currently limits their use in the peak strength condition. Understanding the mechanism of EAC will facilitate the development of crack-resistant alloys with optimum mechanical properties. One component towards understanding the fundamental processes responsible for EAC is a comprehensive knowledge of the chemical conditions within cracks. The present work uses capillary electrophoresis (CE) to quantify the crack chemistry in order to provide insight into the nature of the mechanism controlling cracking. The highly restricted geometry of cracks in metals means that a crack typically contains less than 10 microliters of solution. The high mass sensitivity combined with the inherently robust nature of CE makes it an ideal analytical technique for this application. Complicating factors in the accurate determination of the crack environment include high levels of sodium present from the test solution. Low sample volume and analyte matrix complexity necessitated the development of specific sampling, extraction and analysis methods. Analysis of the crack solutions in EAC-susceptible material revealed high levels of Al3+, Mg2+, Zn2+, and Cl- near the crack tip. Cations arise from the anodic dissolution of the alloy, whereas chloride ingress from the external environment occurs to maintain solution electroneutrality within the crack. In contrast, EAC-resistant material exhibited significantly lower concentrations of dissolution products. PMID:10457501

  9. High-speed DNA genotyping using microfabricated capillary array electrophoresis chips

    Energy Technology Data Exchange (ETDEWEB)

    Woolley, A.T.; Sensabaugh, G.F.; Mathies, R.A. [Univ. of California, Berkeley, CA (United States)

    1997-06-01

    Capillary array electrophoresis (CAE) chips have been designed and fabricated with the capacity to rapidly (<160 s) analyze 12 different samples in parallel. Detection of all lanes with 0.3 s temporal resolution was achieved using a laser-excited confocal-fluorescence scanner. The operation and capabilities of these CAE microdevices were first determined by performing electrophoretic separations of pBR322 MspI DNA samples. Genotyping of HLA-H, a candidate gene for the diagnosis of hereditary hemochromatosis, was then performed to demonstrate the rapid analysis of biologically relevant samples. Two-color multiplex fluorescence detection of HLA-H genotypes was accomplished by prelabeling the standard pBR322 MspI DNA ladder with a red emitting bisintercalation dye (butyl TOTIN) and on-column labeling of the HLA-H DNA with thiazole orange. This work establishes the feasibility of using CAE chips for high-speed, high-throughput genotyping. 44 refs., 7 figs.

  10. Postcolumn derivatization of proteins in capillary sieving electrophoresis/laser-induced fluorescence detection.

    Science.gov (United States)

    Kaneta, Takashi; Yamamoto, Daisuke; Imasaka, Totaro

    2009-11-01

    The separation methods for proteins with high resolution and sensitivity are absolutely important in the field of biological sciences. Capillary sieving electrophoresis (CSE) is an excellent separation technique for DNA and proteins with high resolution, while LIF permits the most sensitive detection in CSE. Therefore, proteins have to be labeled with fluorescent or fluorogenic reagent to produce fluorescent derivatives. Both precolumn and oncolumn derivatization have been employed for the labeling of proteins in CSE. However, there is no report on the postcolumn derivatization due to the limitation in the use of a standard migration buffer, despite it being a promising method for sensitive detection of proteins. Here, we show a novel postcolumn derivatization method for protein separation by CSE, using a tertiary amine as a buffer component in the running buffer. Tris, which is commonly used as a base in CSE separation buffers, was substituted by tertiary amines, 2-(diethylamino)ethanol and triethanolamine. A buffer solution containing 2-(diethylamino)ethanol or triethanolamine can be used for the CSE separation followed by the postcolumn derivatization of proteins, since both reagents are unreactive toward a fluorogenic labeling reagent, naphthalene-2,3-dicarbaldehyde. Thus, LIF detection using the postcolumn derivatization permits significant reduction in the LOD (by a factor of 2.4-28) of proteins, compared with conventional absorbance detection. PMID:19862753

  11. Determination of NTBC in serum samples from patients with hereditary tyrosinemia type I by capillary electrophoresis.

    Science.gov (United States)

    Cansever, M Serif; Aktuğlu-Zeybek, A Ciğdem; Erim, F Bedia

    2010-03-15

    Hereditary tyrosinemia type I is a serious metabolic disorder leading to liver failure. 2-(2-Nitro-4-trifluoromethylbenzoyl)-1,3-cyclohexanedione (NTBC) is a relatively new drug which is used to prevent the accumulation of toxic metabolites in patients with hereditary tyrosinemia type I. In the present study, we have developed a new, simple, fast, and cost-effective capillary electrophoresis method for the quantitative monitoring of this drug in serum samples. Micellar electrochromatographic separation of NTBC was performed using 20 mmol/L phosphate and 40 mmol/L sodium dodecylsulfate (SDS) at pH 12 as running electrolyte. Separation of NTBC was achieved in around 4 min. Reproducibilities of migration times and corrected peak areas of NTBC (as R.S.D.%) were found as 0.73 and 1.99, respectively. The detection limit was 3.17 and the quantification limit was 10.6 micromol/L for NTBC using UV detection at 278 nm. The utility of the method was demonstrated by the detection of NTBC in serum samples from patients with hereditary tyrosinemia type I using this drug. PMID:20152421

  12. Capillary electrophoresis fingerprinting and spectrophotometric determination of antioxidant potential for classification of Mentha products.

    Science.gov (United States)

    Roblová, Vendula; Bittová, Miroslava; Kubáň, Petr; Kubáň, Vlastimil

    2016-07-01

    In this work aqueous infusions from ten Mentha herbal samples (four different Mentha species and six hybrids of Mentha x piperita) and 20 different peppermint teas were screened by capillary electrophoresis with UV detection. The fingerprint separation was accomplished in a 25 mM borate background electrolyte with 10% methanol at pH 9.3. The total polyphenolic content in the extracts was determined spectrophotometrically at 765 nm by a Folin-Ciocalteu phenol assay. Total antioxidant activity was determined by scavenging of 2,2-diphenyl-1-picrylhydrazyl radical at 515 nm. The peak areas of 12 dominant peaks from CE analysis, present in all samples, and the value of total polyphenolic content and total antioxidant activity obtained by spectrophotometry was combined into a single data matrix and principal component analysis was applied. The obtained principal component analysis model resulted in distinct clusters of Mentha and peppermint tea samples distinguishing the samples according to their potential protective antioxidant effect. Principal component analysis, using a non-targeted approach with no need for compound identification, was found as a new promising tool for the screening of herbal tea products.

  13. Metabolomics, peptidomics and proteomics applications of capillary electrophoresis-mass spectrometry in Foodomics: A review

    International Nuclear Information System (INIS)

    Graphical abstract: -- Highlights: •Foodomics allows studying food and nutrition through the application of advanced omics approaches. •CE-MS plays a crucial role as analytical platform to carry out omics studies. •CE-MS applications for food metabolomics, proteomics and peptidomics are presented. -- Abstract: In the current post-genomic era, Foodomics has been defined as a discipline that studies food and nutrition through the application of advanced omics approaches. Foodomics involves the use of genomics, transcriptomics, epigenetics, proteomics, peptidomics, and/or metabolomics to investigate food quality, safety, traceability and bioactivity. In this context, capillary electrophoresis-mass spectrometry (CE-MS) has been applied mainly in food proteomics, peptidomics and metabolomics. The aim of this review work is to present an overview of the most recent developments and applications of CE-MS as analytical platform for Foodomics, covering the relevant works published from 2008 to 2012. The review provides also information about the integration of several omics approaches in the new Foodomics field

  14. Stereoselective analysis of herbicides by capillary electrophoresis using sulfobutyl ether beta-cyclodextrin as chiral selector.

    Science.gov (United States)

    Desiderio, C; Polcaro, C M; Fanali, S

    1997-02-01

    Capillary zone electrophoresis has been used for the enantiomeric separation of several herbicides. Different beta-cyclodextrin (CD) derivatives have been investigated for chiral separations and among them the negatively charged sulfobutyl ether beta-cyclodextrin (SBE-beta-CD) proved to be effective for the stereo-selective resolutions of the investigated herbicides. The effect of CD concentration, buffer pH and organic modifier on effective mobilities, resolution and selectivity of the analytes have been studied. Addition of SBE-beta-CD (5-50 mg/mL) to the buffer at pH 9 resulted in a general increase of migration times as well as resolution. A CD concentration as low as 5 mg/mL was effective to completely resolve napropamide and ethofumesate enantiomers. Buffer solutions containing 40 mg/mL of SBE-beta-CD were chosen to study the effect of buffer pH (7, 8, and 9) on chiral separation of the herbicides. No great differences in resolution and effective mobilities have been found in the pH 7-9 range. The addition of different organic modifiers to the background electrolyte at pH 9, containing 20 mg/mL of SBE-beta-CD, showed different effects. Methanol was the most effective in improving resolution but in some cases total loss in enantiomeric separation was observed. The qualitative analysis of an enantiomerically pure herbicide (flamprop isopropyl) commercial preparation is also shown. PMID:9080130

  15. Simultaneous determination of phenylurea herbicides in yam by capillary electrophoresis with electrochemiluminescence detection.

    Science.gov (United States)

    Hu, Yuefang

    2015-04-01

    A method of capillary electrophoresis (CE) coupled with electrochemiluminescence (ECL) detection has been applied to detect three major phenylurea herbicides (monuron, monolinuron and diuron) simultaneously. The effects of yam sample preparation, injection voltage and time, detection potential, detection buffer concentration and pH, Ru(bpy)₃(2+) concentration, separation buffer type, separation buffer pH and concentration, separation voltage were investigated in detail. Under optimum conditions, a good baseline separation and highly sensitive detection for monuron, monolinuron and diuron were achieved. The ECL intensity (I) was in proportion to three analytes concentration (ρ) in the range of 0.1-10,000 μg/L for monuron (r≥0.9993), 0.1-18,000 μg/L for monolinuron (r≥0.9995) and 0.1-20,000 μg/L for diuron (r≥0.9997). The detection limits for monuron, monolinuron and diuron were 0.05, 0.04 and 0.01 μg/L (S/N=3), respectively. The developed method was successfully applied for the analysis of monuron, monolinuron and diuron residues in yam simultaneously. The average recoveries are in the ranges of 90.0-99.2% with relative standard deviations less than 3.2%. The limits of detection of the proposed method were 0.010 μg/kg for monuron, 0.008 μg/kg for monolinuron and diuron in yam. PMID:25732033

  16. Screening of the binding properties of molecularly imprinted nanoparticles via capillary electrophoresis.

    Science.gov (United States)

    Musile, Giacomo; Cenci, Lucia; Andreetto, Erika; Ambrosi, Emmanuele; Tagliaro, Franco; Bossi, Alessandra Maria

    2016-05-01

    In response to the need for straightforward analytical methods to assess the affinity of molecularly imprinted nanoparticles (MIP NPs) for ligands, capillary electrophoresis (CE) was exploited using MIP NPs targeting the iron-regulating hormone hepcidin. In this work, MIP NPs were challenged with their template peptide, i.e., the N-terminal 5-mer of hepcidin, in comparison to unrelated ligand peptides. A CE separation method was developed ex novo achieving, after optimization of the background electrolyte (150 mM sodium phosphate pH 7.4) and of the running temperature (35 °C), the full separation of the free ligand from the complexed MIP NPs. The CE binding isotherm allowed the estimation of a micromolar dissociation constant for the 5-mer template-MIP NPs complex, in agreement with independent measurements. The CE offered the advantages of a direct injection of the MIP NPs/ligand incubation mix, without preliminary fractionation steps, requiring only minimal sample volumes and short analysis times. In conclusion CE proved to be a valid technique for characterizing the interactions of MIP NP libraries for selected target compounds. Graphical Abstract Five different nanodiamond samples were exhaustively characterized using a suite of analytical techniques. PMID:26960903

  17. Development of a simplified microfluidic injector for analysis of droplet content via capillary electrophoresis.

    Science.gov (United States)

    DeLaMarre, Michael F; Shippy, Scott A

    2014-10-21

    Droplet-based microfluidic platforms sequester nanoliter to picoliter samples in an immiscible carrier phase and have gained notoriety for their ability to be used in laboratory procedures on a miniaturized scale. Recently, droplet microfluidics has been used to prevent zone diffusion in time-resolved sample collection methods and in separation techniques. The assay of droplets remains challenging, however, because the carrier phase is often incompatible with separation techniques. In this work, we report the development of a droplet injector for capillary electrophoresis (CE) which delivers 750 pL droplets to a channel for separation while excluding the fluorous carrier phase. This design is simple compared to previous reports, consisting of only two straight channels and no additional working parts such as membranes or valves. To demonstrate a proof-of-concept and characterize performance, riboflavin was used as a biologically relevant model molecule. Droplets containing a step change in riboflavin concentration were injected and mobilized by CE. The current method is capable of riboflavin peak % relative standard deviations (RSDs) down to 4.4% and temporal resolutions down to 15 s. Human urine samples containing riboflavin and its photolysis products were successfully separated and found to be chemically compatible with the injector. Our simplified design could improve robustness and ruggedness and may allow device construction via nontraditional fabrication techniques. PMID:25226066

  18. Enantiodifferentiation of chiral baclofen by β-cyclodextrin using capillary electrophoresis: A molecular modeling approach

    Science.gov (United States)

    Suliman, FakhrEldin O.; Elbashir, Abdalla A.

    2012-07-01

    Using capillary electrophoresis baclofen (BF) enantiomers were separated only in the presence of β-cyclodextrin (βCD) as a chiral selector when added to the background electrolyte. Proton nuclear magnetic resonance and electrospray ionization mass spectrometry (ESI-MS) techniques were used to determine the structure of the BF-βCD inclusion complexes. From the MS data BF was found to form a 1:1 complex with α- and βCD, while the NMR data suggest location of the aromatic ring of BF into the cyclodextrin cavity. A molecular modeling study, using the semiempirical PM6 calculations was used to investigate the mechanism of enantiodifferentiation of BF with cyclodextrins. Optimization of the structures of the complexes by PM6 method indicated that separation is obtained in the presence of β-CD due to a large binding energy difference (ΔΔE) of 46.8 kJ mol-1 between S-BF-βCD and R-BF-βCD complexes. In the case of αCD complexes ΔΔE was 1.3 kJ mol-1 indicating poor resolution between the two enantiomers. Furthermore, molecular dynamic simulations show that the formation of more stable S-BF-βCD complex compared to R-BF-β-CD complex is primarily due to differences in intermolecular hydrogen bonding.

  19. Identification of primary amines in Titan tholins using microchip nonaqueous capillary electrophoresis

    Science.gov (United States)

    Cable, M. L.; Hörst, S. M.; He, C.; Stockton, A. M.; Mora, M. F.; Tolbert, M. A.; Smith, M. A.; Willis, P. A.

    2014-10-01

    Titan, the moon of Saturn with a thick atmosphere and an active hydrocarbon-based weather cycle, is considered the best target in the solar system for the study of organic chemistry on a planetary scale. Microfluidic devices that employ liquid phase techniques such as capillary electrophoresis with ultrasensitive laser-induced fluorescence detection offer a unique solution for in situ analysis of complex organics on Titan. We previously reported a protocol for nonaqueous microfluidic analysis of primary aliphatic amines in ethanol, and demonstrated separations of short- and long-chain amines down to -20 °C. We have optimized this protocol further, and used it to analyze Titan aerosol analogues (tholins) generated in two separate laboratories under a variety of different conditions. Ethylamine was a major product in all samples, though significant differences in amine content were observed, in particular for long-chain amines (C12-C27). This work validates microfluidic chemical analysis of complex organics with relevance to Titan, and represents a significant first step in understanding tholin composition via targeted functional group analysis.

  20. Simultaneous analysis of saturated and unsaturated fatty acids present in pequi fruits by capillary electrophoresis

    Directory of Open Access Journals (Sweden)

    Patrícia M. de Castro Barra

    2013-01-01

    Full Text Available In the current study, an alternative method has been proposed for simultaneous analysis of palmitic, stearic, oleic, linoleic, and linolenic acids by capillary zone electrophoresis (CZE using indirect detection. The background electrolyte (BGE used for the analysis of these fatty acids (FAs consisted of 15.0 mmol L−1 NaH2PO4/Na2HPO4 at pH 6.86, 4.0 mmol L−1 SDBS, 8.3 mmol L−1 Brij 35, 45% v/v acetonitrile (can, and 2.1% n-octanol. The FAs quantification of FAs was performed using a response factor approach, which provided a high analytical throughput for the real sample. The CZE method, which was applied successfully for the analysis of pequi pulp, has advantages such as short analysis time, absence of lipid fraction extraction and derivatization steps, and no significant difference in the 95% confidence intervals for FA quantification results, compared to the gas chromatography official method (AOCS Ce 1h-05.

  1. Review on the development of truly portable and in-situ capillary electrophoresis systems

    Science.gov (United States)

    Lewis, A. P.; Cranny, A.; Harris, N. R.; Green, N. G.; Wharton, J. A.; Wood, R. J. K.; Stokes, K. R.

    2013-04-01

    Capillary electrophoresis (CE) is a technique which uses an electric field to separate a mixed sample into its constituents. Portable CE systems enable this powerful analysis technique to be used in the field. Many of the challenges for portable systems are similar to those of autonomous in-situ analysis and therefore portable systems may be considered a stepping stone towards autonomous in-situ analysis. CE is widely used for biological and chemical analysis and example applications include: water quality analysis; drug development and quality control; proteomics and DNA analysis; counter-terrorism (explosive material identification) and corrosion monitoring. The technique is often limited to laboratory use, since it requires large electric fields, sensitive detection systems and fluidic control systems. All of these place restrictions in terms of: size, weight, cost, choice of operating solutions, choice of fabrication materials, electrical power and lifetime. In this review we bring together and critique the work by researchers addressing these issues. We emphasize the importance of a holistic approach for portable and in-situ CE systems and discuss all the aspects of the design. We identify gaps in the literature which require attention for the realization of both truly portable and in-situ CE systems.

  2. Microfluidic-based metal enhanced fluorescence for capillary electrophoresis by Ag nanorod arrays

    Science.gov (United States)

    Xiao, Chenyu; Cao, Zhen; Deng, Junhong; Huang, Zhifeng; Xu, Zheng; Fu, Junxue; Yobas, Levent

    2014-06-01

    As metal nanorods show much higher metal enhanced fluorescence (MEF) than metal nanospheres, microfluidic-based MEF is first explored with Ag nanorod (ND) arrays made by oblique angle deposition. By measuring the fluorescein isothiocyanate (FITC) solution sandwiched between the Ag NDs and a piece of cover slip, the enhancement factors (EFs) are found as 3.7 ± 0.64 and 6.74 ± 2.04, for a solution thickness at 20.8 μm and 10 μm, respectively. Because of the strong plasmonic coupling between the adjacent Ag NDs, only the emission of the fluorophores present in the three-dimensional NDs array gets enhanced. Thus, the corresponding effective enhancement factors (EEFs) are revealed to be relatively close, 259 ± 92 and 340 ± 102, respectively. To demonstrate the application of MEF in microfluidic systems, a multilayer of SiO2 NDs/Ag NDs is integrated with a capillary electrophoresis device. At a microchannel depth of 10 μm, an enhancement of 6.5 fold is obtained for amino acids separation detection. These results are very encouraging and open the possibility of MEF applications for the Ag ND arrays decorated microchannels. With the miniaturization of microfluidic devices, microfluidic-based MEF by Ag ND arrays will likely find more applications with further enhancement.

  3. Capillary electrophoresis fingerprinting and spectrophotometric determination of antioxidant potential for classification of Mentha products.

    Science.gov (United States)

    Roblová, Vendula; Bittová, Miroslava; Kubáň, Petr; Kubáň, Vlastimil

    2016-07-01

    In this work aqueous infusions from ten Mentha herbal samples (four different Mentha species and six hybrids of Mentha x piperita) and 20 different peppermint teas were screened by capillary electrophoresis with UV detection. The fingerprint separation was accomplished in a 25 mM borate background electrolyte with 10% methanol at pH 9.3. The total polyphenolic content in the extracts was determined spectrophotometrically at 765 nm by a Folin-Ciocalteu phenol assay. Total antioxidant activity was determined by scavenging of 2,2-diphenyl-1-picrylhydrazyl radical at 515 nm. The peak areas of 12 dominant peaks from CE analysis, present in all samples, and the value of total polyphenolic content and total antioxidant activity obtained by spectrophotometry was combined into a single data matrix and principal component analysis was applied. The obtained principal component analysis model resulted in distinct clusters of Mentha and peppermint tea samples distinguishing the samples according to their potential protective antioxidant effect. Principal component analysis, using a non-targeted approach with no need for compound identification, was found as a new promising tool for the screening of herbal tea products. PMID:27306863

  4. [Simultaneous separation and determination of vanillin and o-vanillin by capillary zone electrophoresis].

    Science.gov (United States)

    Chen, Xing; Guan, Jin; Wang, Huize; Li, Yun; Shi, Zhe

    2010-11-01

    A method for the simultaneous separation and determination of vanillin and o-vanillin by capillary zone electrophoresis (CZE) was developed. The influences of type, concentration and pH of running buffer, and applied voltage on separation were investigated. Under the conditions of 50 mmol/L borax-150 mmol/L disodium hydrogen phosphate (pH 7.5) and applied voltage of 15 kV, the vanillin and o-vanillin were separated in 6 min. The method was proved to be robust through verification of accuracy, precision and linearity. The calibration curves of vanillin and o-vanillin showed good linearity in the range of 10-240 mg/L, and the correlation coefficients were 0.999 9 and 0.999 7, respectively. The limits of detection for vanillin and o-vanillin were 1.0 mg/L (S/N = 3). The average recoveries at three spiked levels were 99.4%-101.2% with acceptable relative standard deviations of 0.19%-0.73%. The method has been successfully used for the determination of vanillin and o-vanillin in real samples, and the assay results are satisfactory.

  5. End-to-end differential contactless conductivity sensor for microchip capillary electrophoresis.

    Science.gov (United States)

    Fercher, Georg; Haller, Anna; Smetana, Walter; Vellekoop, Michael J

    2010-04-15

    In this contribution, a novel measurement approach for miniaturized capillary electrophoresis (CE) devices is presented: End-to-end differential capacitively coupled contactless conductivity measurement. This measurement technique is applied to a miniaturized CE device fabricated in low-temperature cofired ceramics (LTCC) multilayer technology. The working principle is based on the placement of two distinct detector areas near both ends of the fluid inlet and outlet of the separation channel. Both output signals are subtracted from each other, and the resulting differential signal is amplified and measured. This measurement approach has several advantages over established, single-end detectors: The high baseline level resulting from parasitic stray capacitance and buffer conductivity is reduced, leading to better signal-to-noise ratio and hence higher measurement sensitivity. Furthermore, temperature and, thus, baseline drift effects are diminished owing to the differentiating nature of the system. By comparing the peak widths measured with both detectors, valuable information about zone dispersion effects arising during the separation is obtained. Additionally, the novel measurement scheme allows the determination of dispersion effects that occur at the time of sample injection. Optical means of dispersion evaluation are ineffective because of the opaque LTCC substrate. Electrophoretic separation experiments of inorganic ions show sensitivity enhancements by about a factor of 30-60 compared to the single-end measurement scheme. PMID:20337422

  6. Capillary electrophoresis to determine entrapment efficiency of a nanostructured lipid carrier loaded with piroxicam

    Institute of Scientific and Technical Information of China (English)

    Jessica Otarola; Adriana Guillermina Lista; Beatriz Fernández Band; Mariano Garrido

    2015-01-01

    A simple and fast capillary electrophoresis method has been developed to determine the amount of piroxicam loaded in a drug delivery system based on nanostructured lipid carriers (NLCs). The entrapment efficiency of the nanostructured lipid carrier was estimated by measuring the concentration of drug not entrapped in a suspension of NLC. The influence of different parameters on migration times, peak symmetry, efficiency and resolution was studied; these parameters included the pH of the electrophoretic buffer solution and the applied voltage. The piroxicam peak was obtained with a satisfactory resolution. The separation was carried out using a running buffer composed of 50 mM ammonium acetate and 13.75 mM ammonia at pH 9. The optimal voltage was 20 kV and the cartridge temperature was 20 1C. The corresponding calibration curve was linear over the range of 2.7–5.4 mg/mL of NLC suspension. The reproducibility of migration time and peak area were investigated, and the obtained RSD% values (n ¼ 5) were 0.99 and 2.13, respectively.

  7. Capillary electrophoresis-based immobilized enzyme reactor using particle-packing technique.

    Science.gov (United States)

    Liu, Lina; Zhang, Bo; Zhang, Qian; Shi, Yanhong; Guo, Liping; Yang, Li

    2014-07-25

    A novel method using particle-packing technique to fabricate capillary electrophoresis (CE)-based immobilized enzyme reactor (IMER) was accomplished by utilizing perfusive silica single particles as the frits and large-pore beads as the enzyme supports. The fabrication procedure is rapid and simple; the length and enzyme loading amount of the CE-IMERs could be easily adjusted. Performance and feasibility of the CE-IMERs were investigated using on-line trypsin digestion as the model enzyme reaction. High reproducible on-line enzyme assay was demonstrated with RSD less than 4.1% and 3.8% for peak area and migration time of the substrate and product over 100 consecutive runs, respectively. The enzyme can still maintain the activity for at least 10 days, indicating remarkably stability of the CE-IMERs. The CE-IMERs were successfully applied for accurate analysis of trypsin inhibition as well as on-line digestion of standard proteins (myoglobin and BSA). The present method provides a new interesting alternative to open-tubular and monolithic CE-IMERs, thus expands the application of the CE technique for on-line enzyme assay and analysis and characterization of peptides and proteins.

  8. Centimeter-scale characterization of biogeochemical gradients at a wetland-aquifer interface using capillary electrophoresis

    Science.gov (United States)

    Baez-Cazull, S.; McGuire, J.T.; Cozzarelli, I.M.; Raymond, A.; Welsh, L.

    2007-01-01

    Steep biogeochemical gradients were measured at mixing interfaces in a wetland-aquifer system impacted by landfill leachate in Norman, Oklahoma. The system lies within a reworked alluvial plain and is characterized by layered low hydraulic conductivity wetland sediments interbedded with sandy aquifer material. Using cm-scale passive diffusion samplers, "peepers", water samples were collected in a depth profile to span interfaces between surface water and a sequence of deeper sedimentary layers. Geochemical indicators including electron acceptors, low-molecular-weight organic acids, base cations, and NH4+ were analyzed by capillary electrophoresis (CE) and field techniques to maximize the small sample volumes available from the centimeter-scale peepers. Steep concentration gradients of biogeochemical indicators were observed at various interfaces including those created at sedimentary boundaries and boundaries created by heterogeneities in organic C and available electron acceptors. At the sediment-water interface, chemical profiles with depth suggest that SO42 - and Fe reduction dominate driven by inputs of organic C from the wetland and availability of electron acceptors. Deeper in the sediments (not associated with a lithologic boundary), a steep gradient of organic acids (acetate maximum 8.8 mM) and NH4+ (maximum 36 mM) is observed due to a localized source of organic matter coupled with the lack of electron acceptor inputs. These findings highlight the importance of quantifying the redox reactions occurring in small interface zones and assessing their role on biogeochemical cycling at the system scale. ?? 2007 Elsevier Ltd. All rights reserved.

  9. Comparative study between capillary electrophoresis and high performance liquid chromatography in 'guarana' based phytopharmaceuticals.

    Science.gov (United States)

    Sombra, Lorena L; Gómez, María R; Olsina, Roberto; Martínez, Luis D; Silva, María F

    2005-01-01

    The last years have seen a significant increase in the use of herbal medicines and their preparations all over the world. Adulterations with synthetic drugs are common problems with phytopharmaceutical products and this can potentially cause adverse effects. In consequence, it is important to determine the presence of synthetic drugs in herbal medicines to ensure their efficacy and safety. In this study, guarana derivatives were analyzed by capillary electrophoresis (CE), and the results were compared with those obtained by the HPLC technique. In order to obtain adequate fingerprints, and search for adulterants, caffeine was used as the marker compound. This separation method was applied to analyze the seed powder and commercial tablets of Paulinia cupana Mart. The methodology performance was evaluated in terms of specificity, sensitivity and precision. The results are in agreement with those obtained by the HPLC method. Furthermore, the analysis time of the CE method is up to two times shorter than the respective parameter in HPLC and solvent consumption is more than 100-fold less.

  10. Determination of low molecular weight organic acids in soil, plants, and water by capillary zone electrophoresis.

    Science.gov (United States)

    Li, Ying-Hui; Huang, Bi-Xia; Shan, Xiao-Quan

    2003-03-01

    Determination of low molecular weight organic acids in soils and plants by capillary zone electrophoresis was accomplished using a phthalate buffer and indirect UV detection mode. The influence of some crucial parameters, such as pH, buffer concentration and surfactant were investigated. A good separation of seven organic acids was achieved within 5 min using an electrolyte containing 15 mmol L(-1) potassium hydrogen phthalate, 0.5 mmol L(-1) myristyltrimethylammonium bromide (MTAB), and 5% methanol (MeOH) (v/v) at pH 5.60, separation voltage -20 kV, and temperature 25 degrees C. The relative standard deviation (n=5) of the method was found to be in range 0.18-0.56% for migration time and 3.2-4.8% for peak area. The limit of detection ranged between 0.5 micro mol L(-1) to 6 micro mol L(-1) at a signal-to-noise ratio of 3. The recovery of standard organic acids added to real samples ranged from 87 to 119%. This method was simple, rapid and reproducible, and could be applied to the simultaneous determination of organic acids in environmental samples. PMID:12664177

  11. Simultaneous Detection of Genetically Modified Organisms in a Mixture by Multiplex PCR-Chip Capillary Electrophoresis.

    Science.gov (United States)

    Patwardhan, Supriya; Dasari, Srikanth; Bhagavatula, Krishna; Mueller, Steffen; Deepak, Saligrama Adavigowda; Ghosh, Sudip; Basak, Sanjay

    2015-01-01

    An efficient PCR-based method to trace genetically modified food and feed products is in demand due to regulatory requirements and contaminant issues in India. However, post-PCR detection with conventional methods has limited sensitivity in amplicon separation that is crucial in multiplexing. The study aimed to develop a sensitive post-PCR detection method by using PCR-chip capillary electrophoresis (PCR-CCE) to detect and identify specific genetically modified organisms in their genomic DNA mixture by targeting event-specific nucleotide sequences. Using the PCR-CCE approach, novel multiplex methods were developed to detect MON531 cotton, EH 92-527-1 potato, Bt176 maize, GT73 canola, or GA21 maize simultaneously when their genomic DNAs in mixtures were amplified using their primer mixture. The repeatability RSD (RSDr) of the peak migration time was 0.06 and 3.88% for the MON531 and Bt176, respectively. The RSD (RSDR) of the Cry1Ac peak ranged from 0.12 to 0.40% in multiplex methods. The method was sensitive in resolving amplicon of size difference up to 4 bp. The PCR-CCE method is suitable to detect multiple genetically modified events in a composite DNA sample by tagging their event specific sequences. PMID:26525256

  12. Determination of Nicotine in Tobacco by Capillary Electrophoresis with Electrochemical Detection

    Institute of Scientific and Technical Information of China (English)

    SUN Jin-ying; XU Xiao-yu; YU Huan; YOU Tian-yan

    2012-01-01

    A sensitive,simple and low-cost method based on capillary electrophoresis(CE) with electrochemical(EC) detection at a carbon fiber microdisk electrode(CFE) was developed for the determination of nicotine.Effects of detection potential,concentration and pH value of the phosphate buffer,and injection time as well as separation voltage were investigated.Under the optimized conditions:a detection potential of 1.20 V,40 mmol/L phosphate buffer(pH 2.0),a sample injection time of 10 s at 10 kV and a separation voltage of 16 kV,the linear range obtained was from 5.0× 10 7 mol/L to 1.0× 10-4 mol/L with a correlation coefficient of 0.9989 and the limit of detection(LOD,S/N=3)obtained was 5.0× 10-8 mol/L.The method was also used to determine the nicotine in cigarettes.Nicotine amount ranged from 0.211 mg/g to 0.583 mg/g in the pipe tobacco of seven brands of cigarette and the amount in one cigarette varied from 0.136 mg/cigarette to 0.428 mg/cigarette.

  13. Quality Analysis of Herbal Medicine Products Prepared from Herba Sarcandrae by Capillary Electrophoresis with Electrochemical Detection

    Institute of Scientific and Technical Information of China (English)

    ZHOU Xiao-guang; SUN Jin-ying; ZHU De-rong; YUAN Bai-qing; YOU Tian-yan

    2008-01-01

    A capillary electrophoresis with electrochemical detection(CE-ED)method was developed for the quality analysis of herbal medicine products prepared from the sanle herb of Herba Sarcandrae:Fufang Caoshanhu tablets,Qingrexiaoyanning capsules,and Xuekang oral liquids.Under the optimal analysis conditions,the low detection limit[1.0×10-7mol/L(S/N=3)]and the wide linear range(1.0×10-7-1.0×10-4 mol/L)were obtained for quality standard compound of isofraxidin.The precisions of the peak current and the migration time(as RSDs)for the real sample analysis were 2.0%-2.6%,and 1.2%-1.8%for isofraxidin,respectively.The contents of isofraxidin detected were 15.77 μg/tablet,0.48 mg/capsule,1.2 mg/ampoule(Jiangxi),and 0.44 mg/ampoule(Dalian)for Fufang Canshanhu tablets,Qingrexiao yanning capsules,and Xuekang oral liquids from different manufacturers,respectively.Quality estimate Was conducted by comparing the contents of isofraxidin in the herbal medicine products with the demanded values of Chinese pharmacopeia.In addition,based on their own unique CE-ED profiles(namely,CE-ED electropherograrns)the Xuekang oral liquids from the different manufacturers could be easily identified.

  14. Capillary electrophoresis method with UV-detection for analysis of free amino acids concentrations in food.

    Science.gov (United States)

    Omar, Mei Musa Ali; Elbashir, Abdalla Ahmed; Schmitz, Oliver J

    2017-01-01

    Simple and inexpensive capillary electrophoresis with UV-detection method (CE-UV) was optimized and validated for determination of six amino acids namely (alanine, asparagine, glutamine, proline, serine and valine) for Sudanese food. Amino acids in the samples were derivatized with 4-chloro-7-nitro-2,1,3-benzoxadiazole (NBD-Cl) prior to CE-UV analysis. Labeling reaction conditions (100mM borate buffer at pH 8.5, labeling reaction time 60min, temperature 70°C and NBD-Cl concentration 40mM) were systematically investigated. The optimal conditions for the separation were 100mM borate buffer at pH 9.7 and detected at 475nm. The method was validated in terms of linearity, limit of detection (LOD), limit of quantification (LOQ), precision (repeatability) (RSD%) and accuracy (recovery). Good linearity was achieved for all amino acids (r(2)>0.9981) in the concentration range of 2.5-40mg/L. The LODs in the range of 0.32-0.56mg/L were obtained. Recoveries of amino acids ranging from 85% to 108%, (n=3) were obtained. The validated method was successfully applied for the determination of amino acids for Sudanese food samples. PMID:27507479

  15. [Selection of back-ground electrolyte in capillary zone electrophoresis by triangle and tetrahedron optimization methods].

    Science.gov (United States)

    Sun, Guoxiang; Song, Wenjing; Lin, Ting

    2008-03-01

    The triangle and tetrahedron optimization methods were developed for the selection of back-ground electrolyte (BGE) in capillary zone electrophoresis (CZE). Chromatographic fingerprint index F and chromatographic fingerprint relative index F(r) were used as the objective functions for the evaluation, and the extract of Saussurea involucrate by water was used as the sample. The BGE was composed of borax, boric acid, dibasic sodium phosphate and sodium dihydrogen phosphate solution with different concentrations using triangle and tetrahedron optimization methods. Re-optimization was carried out by adding organic modifier to the BGE and adjusting the pH value. In triangle method, when 50 mmol/L borax-150 mmol/L sodium dihydrogen phosphate (containing 3% acetonitrile) (1 : 1, v/v) was used as BGE, the isolation was considered to be satisfactory. In tetrahedron method, the best BGE was 50 mmol/L borax-150 mmol/L sodium dihydrogen phosphate-200 mmol/L boric acid (1 : 1 : 2, v/v/v; adjusting the pH value to 8.55 by 0.1 mol/L sodium hydroxide). There were 28 peaks and 25 peaks under the different conditions respectively. The results showed that the methods could be applied to the selection of BGE in CZE of the extract of traditional Chinese medicine by water or ethanol.

  16. Synthesis of ino Acid Derived β-Cyclodextrins Used in Chiral Separation by Capillary Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    戴荣继; 佟斌; 魏征; 顾峻岭; 邓玉林; 李明愉; 傅若农

    2004-01-01

    Six new kinds of ino acid derived β-cyclodextrins were synthesized to improve their water solubility and chiral separation properties. They are heptakis{2,6-di-O-[3-L-(1-isopropyl carboxyl methyl ino)-2-hydroxy propyl]}-β-cyclodextrin (i.e. L-Val-β-CD), heptakis{2,6-di-O-[3-L-(1-benzyl carboxyl methyl ino)-2-hydroxy propyl]}-β-cyclodextrin (i.e. L-Phe-β-CD), heptakis{2,6-di-O-[3-(D, L-1-benzyl carboxyl methyl ino)-2-hydroxy propyl]}-β-cyclodextrin (i.e. D,L-Phe-β-CD), heptakis{2,6-di-O-[3-(L-1-hydroxymethyl carboxyl methyl ino)-2-hydroxy propyl]}-β-cyclodextrin (i.e. L-Ser-β-CD), heptakis{2,6-di-O-[3-(L-1-carboxylmethyl carboxyl methyl ino)- 2-hydroxy propyl]}-β-cyclodextrin (i.e. L-Asp-β-CD), heptakis{2,6-di-O-[3-(L-2-carboxyl tetrethylene ino)-2-hydroxy propyl]}-β-cyclodextrin (i.e. L-Pro-β-CD). Their chemical structures were certified using FTIR and 1H NMR. Except for L-Phe-β-CD and D,L-Phe-β-CD, that are in soluble in water, the other ino acid derived β-CDs all have good water solubility. D,L-tyrosine and promethazine were baselinely separated by L-Val-β-CD in capillary electrophoresis.

  17. Methoxypropylamino β-cyclodextrin clicked AC regioisomer for enantioseparations in capillary electrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, Jie; Wang, Yiying; Liu, Yun; Tang, Jian; Tang, Weihua, E-mail: whtang@mail.njust.edu.cn

    2015-04-08

    Highlights: In this paper, we demonstrate: • The click synthesis of a AC regioisomer cationic cyclodextrin (CD) as chiral selector. • The good enantioselectivities (chiral resolution over 5) for acidic racemates. • The strong chiral recognition of new CD by NMR study. • Baseline enantioseparation of some acidic racemates at CD of 0.5 mM. - Abstract: In this work, a novel methoxypropylamino β-cyclodextrin (β-CD) clicked AC regioisomer, 6{sup A}-4-hydroxyethyl-1,2,3-triazolyl-6{sup C}-3-methoxypropylamino β-cyclodextrin (HETz-MPrAMCD), was synthesized via nucleophilic addition and click chemistry. The chiral separation ability of this AC regioisomer cationic CD was evaluated toward 7 ampholytic and 13 acidic racemates by capillary electrophoresis. Dependence of enantioselectivity and resolution on buffer pH (5.5–8.0) and chiral selector concentration (0.5–7.5 mM) was investigated. Enantioselectivities (α ≥ 1.05) could be achieved for most analytes under optimal conditions except dansyl-DL-noreleucine and dansyl-DL-serine. The highest resolutions for 2-chloromandelic acid p-hydroxymandelic acid were 15.6 and 9.7 respectively. The inclusion complexation between HETz-MPrAMCD and each 3-phenyllactic acid enantiomer was also revealed with nuclear magnetic resonance study.

  18. Capillary electrophoresis with electrochemiluminescent detection for highly sensitive assay of genetically modified organisms.

    Science.gov (United States)

    Guo, Longhua; Yang, Huanghao; Qiu, Bin; Xiao, Xueyang; Xue, Linlin; Kim, Donghwan; Chen, Guonan

    2009-12-01

    A capillary electrophoresis coupled with electrochemiluminescent detection system (CE-ECL) was developed for the detection of polymerase chain reaction (PCR) amplicons. The ECL luminophore, tris(1,10-phenanthroline) ruthenium(II) (Ru(phen)(3)(2+)), was labeled to the PCR primers before amplification. Ru(phen)(3)(2+) was then introduced to PCR amplicons by PCR amplification. Eventually, the PCR amplicons were separated and detected by the homemade CE-ECL system. The detection of a typical genetically modified organism (GMO), Roundup Ready Soy (RRS), was shown as an example to demonstrate the reliability of the proposed approach. Four pairs of primers were amplified by multiple PCR (MPCR) simultaneously, three of which were targeted on the specific sequence of exogenous genes of RRS, and another was targeted on the endogenous reference gene of soybean. Both the conditions for PCR amplification and CE-ECL separation and detection were investigated in detail. Results showed that, under the optimal conditions, the proposed method can accurately identifying RRS. The corresponding limit of detection (LOD) was below 0.01% with 35 PCR cycles.

  19. Simultaneous Detection of Genetically Modified Organisms in a Mixture by Multiplex PCR-Chip Capillary Electrophoresis.

    Science.gov (United States)

    Patwardhan, Supriya; Dasari, Srikanth; Bhagavatula, Krishna; Mueller, Steffen; Deepak, Saligrama Adavigowda; Ghosh, Sudip; Basak, Sanjay

    2015-01-01

    An efficient PCR-based method to trace genetically modified food and feed products is in demand due to regulatory requirements and contaminant issues in India. However, post-PCR detection with conventional methods has limited sensitivity in amplicon separation that is crucial in multiplexing. The study aimed to develop a sensitive post-PCR detection method by using PCR-chip capillary electrophoresis (PCR-CCE) to detect and identify specific genetically modified organisms in their genomic DNA mixture by targeting event-specific nucleotide sequences. Using the PCR-CCE approach, novel multiplex methods were developed to detect MON531 cotton, EH 92-527-1 potato, Bt176 maize, GT73 canola, or GA21 maize simultaneously when their genomic DNAs in mixtures were amplified using their primer mixture. The repeatability RSD (RSDr) of the peak migration time was 0.06 and 3.88% for the MON531 and Bt176, respectively. The RSD (RSDR) of the Cry1Ac peak ranged from 0.12 to 0.40% in multiplex methods. The method was sensitive in resolving amplicon of size difference up to 4 bp. The PCR-CCE method is suitable to detect multiple genetically modified events in a composite DNA sample by tagging their event specific sequences.

  20. Simultaneous determination of flavonoids and anthraquinones in chrysanthemum by capillary electrophoresis with amperometry detection

    Institute of Scientific and Technical Information of China (English)

    Yin Yan Zhang; Zi Cheng Li; Jin Kun Zhu; Zhi Yong Yang; Qing Jiang Wang; Pin Gang He; Yu Zhi Fang

    2010-01-01

    A high-performance capillary electrophoresis with amperometry detection method(CE-AD)has been developed for the analysis of flavonoids and anthraquinones(emodin,kaempferol,apigenin,luteolin and rhein)in chrysanthemum.Under optimum conditions,these five analytes were base-line separated within 17 min using a borate-phosphate running buffer(1.5 × 10-2mol/L borate-3 × 10-2 mol/L phosphate running buffer,pH 9.0)at a working potential of+0.90 V(vs.SCE)and a separation voltage of 19 kV.The linear relationship between concentration and current response was obtained with detection limits(S/N = 3)ranging from 1.0 × 10-7 to 2.1 × 10-7 g/mL for all analytes.This proposed method was successfully used in the analysis of four kinds of chrysanthemum with relatively simple extraction procedures,the assay results were satisfactory.

  1. Uranyl complexation with acetate studied by means of affinity capillary electrophoresis.

    Science.gov (United States)

    Sladkov, Vladimir

    2013-05-10

    The interaction of uranyl with acetate is studied by affinity capillary electrophoresis in aqueous acid solutions at the pH values 2.0 and 2.5. The use of data on metal ion mobilities at different pHs allows to establish the ligand species interacting with metal ion and complex species formed. The formation of two complex species UO2CH3COO(+) and UO2(CH3COO)2 is observed (acetic acid concentration is up to 0.8M). In the case of uranyl-acetic acid system, the viscosity of solution is significantly changed with an increase of acid concentration. For calculation of ion mobilities the viscosity changes are taken into account. The stability constants are calculated at the ionic strengths 0.02 and 0.05 mol L(-1). The logarithms of the thermodynamic stability constants (β°) calculated with Davies equation for the activity coefficients of the ions are log β1(°)=2.94±0.08 and log β2(°)=5.50±0.15 at 25 °C. Obtained values are compared with literature data. PMID:23570853

  2. High-sensitivity quantum dot-based fluorescence resonance energy transfer bioanalysis by capillary electrophoresis.

    Science.gov (United States)

    Li, Yong-Qiang; Wang, Jian-Hao; Zhang, Hai-Li; Yang, Jie; Guan, Li-Yun; Chen, Hong; Luo, Qing-Ming; Zhao, Yuan-Di

    2010-02-15

    Here a new method for high-sensitivity quantum dot (QD)-based fluorescence resonance energy transfer (FRET) bioanalysis was developed. In this method, capillary electrophoresis (CE) with fluorescence detection was applied. The FRET system consisted of water-soluble 532-nm emitting CdTe QDs donor and 632-nm emitting CdSe/ZnS QDs acceptor which were covalently conjugated with mouse IgG and goat anti-mouse IgG, respectively. The bio-affinity between antigen and antibody brought two kinds of QDs close enough to make the FRET happen between them. In the CE experiments, highly efficient separation of donor-acceptor immunocomplexes was obtained, and the process of FRET was monitored. Results showed that FRET efficiency obtained by CE (38.56-69.58%) improved substantially in comparison with that obtained by ensemble measurement (12.77-52.37%). The high efficient separation of donor-acceptor immunocomplexes and the possible conformation change of antigen and antibody, contributes to the lower analysis uncertainty (variance) and higher FRET efficiency obtained in CE and consequentially, this makes the analysis of FRET more sensitive. This novel CE-based technique can be easily extended to other FRET system based on QDs and may have potential application in the study of biomolecule conformation change. PMID:19914053

  3. A Capillary Electrophoresis Detection Scheme for Water-soluble Vitamins Based on Luminol - BrO- Chemiluminescence System

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    A novel chemiluminescence detection scheme has been developed for detecting water-soluble vitamins following capillary electrophoresis (CE) separation. This detection was based on the inhibitory effect of vitamins on the CL reaction between luminol and BrO- in basic aqueous solution. Detection of vitamins was accomplished with a borate-based background electrolyte at pH 9.2. The luminol was used as a component of the separation carrier electrolyte.

  4. Further development and applications of capillary electrophoresis with capacitively coupled contactless conductivity detection and sequential injection analysis in analytical chemistry

    OpenAIRE

    Stojkovic, Marko

    2013-01-01

    This dissertation is based on the further development and applications of capillary electrophoresis (CE) with capacitively coupled contactless conductivity detection (C4D), i. e. sequential injection analysis (SIA) applications when coupled with CE-C4D, or determination and quantification of various ions that are not or barely UV absorbed. A purpose made CE-C4D system was used for determination of the DNA fragments of different length, using additives to modify the medium and to sieve ch...

  5. Capillary Zone Electrophoresis Investigation of Interactions between Granulocyte-colony Stimulating Factor and Dextran Sulfate / Carrageenan Oligosaccharide

    Institute of Scientific and Technical Information of China (English)

    Ai Ye LIANG; Yu Guang DU; Ke Yi WANG; Bing Cheng LIN

    2005-01-01

    The interactions between granulocyte-colony stimulating factor (G-CSF) and dextran sulfate / κ-carrageenan oligosa1ccharide were studied by capillary zone electrophoresis. Dextran sulfate could strongly interact with G-CSF and the complex was detected. The binding constant and stoichiometry were determined to be 1.2x106 (mol/L)-1 and 3:1, respectively. However, the interaction between κ-carrageenan oligosaccharide and G-CSF was not found.

  6. Enantiomeric Separation of Antidepressant Trimipramine by Capillary Electrophoresis Combined with Electrochemiluminescence Detection in Aqueous-organic Media

    Institute of Scientific and Technical Information of China (English)

    YU Cai-xia; YUAN Bai-qing; YOU Tian-yan

    2011-01-01

    The antidepressant trimipramine(Tri) enantiomers were successfully separated by capillary electrophoresis(CE) coupled with electrochemiluminescence(ECL) detection in aqueous-organic media. A dual cyclodextrin(CD)system combining β-CD and hydroxypropyl-β-cyclodextrin(HP-β-CD) was used as chiral selector. Acetonitrile(ACN)was added to the running buffer to improve the separation efficiency, detection sensitivity and repeatability. The method was also successfully applied to the chiral separation of Tri in spiked human urine sample.

  7. Elemental speciation by capillary electrophoresis with inductively coupled plasma spectrometry: A new approach by flow focusing® nebulization

    OpenAIRE

    Kovachev, Nikolay; Aguirre Pastor, Miguel Ángel; Hidalgo Núñez, Montserrat; Simitchiev, Kiril; Stefanova, Violeta M.; Kmetov, Veselin Y.; Canals Hernández, Antonio

    2014-01-01

    A novel system for Capillary Electrophoresis (CE) and Inductively Coupled Plasma (ICP) sample introduction that incorporates a dedicated Flow-Focusing® based nebulizer as aerosol generation unit is presented, aiming to provide high signal sensitivity and low detection limits for element speciation at short analysis times. To prove its viability, the system prototype constructed has been coupled to an inductively coupled plasma - optical emission spectrometer (ICP-OES) and an inductively coupl...

  8. Near infrared spectroscopy compared to liquid chromatography coupled to mass spectrometry and capillary electrophoresis as a detection tool for peptide reaction monitoring.

    Science.gov (United States)

    Petter, Christine H; Heigl, Nico; Bachmann, Stefan; Huck-Pezzei, Verena A C; Najam-Ul-Haq, Muhammad; Bakry, Rania; Bernkop-Schnürch, Andreas; Bonn, Günther K; Huck, Christian W

    2008-05-01

    Peptide interaction is normally monitored by liquid chromatography (LC), liquid chromatography coupled to mass spectrometry (LC-MS), mass spectrometric (MS) methods such as MALDI-TOF/MS or capillary electrophoresis (CE). These analytical techniques need to apply either high pressure or high voltages, which can cause cleavage of newly formed bondages. Therefore, near infrared reflectance spectroscopy (NIRS) is presented as a rapid alternative to monitor the interaction of glutathione and oxytocin, simulating physiological conditions. Thereby, glutathione can act as a nucleophile with oxytocin forming four new conjugates via a disulphide bondage. Liquid chromatography coupled to UV (LC-UV) and mass spectrometry via an electrospray ionisation interface (LC-ESI-MS) resulted in a 82% and a 78% degradation of oxytocin at pH 3 and a 5% and a 7% degradation at pH 6.5. Capillary electrophoresis employing UV-detection (CE-UV) showed a 44% degradation of oxytocin. LC and CE in addition to the NIRS are found to be authentic tools for quantitative analysis. Nevertheless, NIRS proved to be highly suitable for the detection of newly formed conjugates after separating them on a thin layer chromatography (TLC) plate. The recorded fingerprint in the near infrared region allows for a selective distinct qualitative identification of conjugates without the need for expensive instrumentation such as quadrupole or MALDI-TOF mass spectrometers. The performance of the established NIRS method is compared to LC and CE; its advantages are discussed in detail. PMID:18095054

  9. One step physically adsorbed coating of silica capillary with excellent stability for the separation of basic proteins by capillary zone electrophoresis.

    Science.gov (United States)

    Guo, Xiao-Feng; Guo, Xiao-Mei; Wang, Hong; Zhang, Hua-Shan

    2015-11-01

    The coating of capillary inner surface is considered to be an effective approach to suppress the adsorption of proteins on capillary inner surface in CE. However, most of coating materials reported are water-soluble, which may dissolve in BGE during the procedure of electrophoresis. In this study, a novel strategy for selection of physically coating materials has been illustrated to get coating layer with excellent stability using materials having poor solubility in commonly used solvents. Taking natural chitin as example (not hydrolyzed water soluble chitosan), a simple one step coating method using chitin solution in hexafluoroisopropanol was adopted within only 21 min with good coating reproducibility (RSDs of EOF for within-batch coated capillaries of 1.55% and between-batch coated capillaries of 2.31%), and a separation of four basic proteins on a chitin coated capillary was performed to evaluate the coating efficacy. Using chitin coating, the adsorption of proteins on capillary inner surface was successfully suppressed with reversed and stable EOF, and four basic proteins including lysozyme, cytochrome c, ribonuclease A and α-chymotrypsinogen A were baseline separated within 16 min with satisfied separation efficiency using 20 mM pH 2.0 H3PO4-Na2HPO4 as back ground electrolyte and 20 kV as separation voltage. What is more important, the chitin coating layer could be stable for more than two months during this study, which demonstrates that chitin is an ideal material for preparing semi-permanent coating on bare fused silica capillary inner wall and has hopeful potential in routine separation of proteins with CE.

  10. Boron-doped diamond microelectrodes for use in capillary electrophoresis with electrochemical detection.

    Science.gov (United States)

    Cvacka, Josef; Quaiserová, Veronika; Park, JinWoo; Show, Yoshiyuki; Muck, Alexander; Swain, Greg M

    2003-06-01

    The fabrication and characterization of boron-doped diamond microelectrodes for use in electrochemical detection coupled with capillary electrophoresis (CE-EC) is discussed. The microelectrodes were prepared by coating thin films of polycrystalline diamond on electrochemically sharpened platinum wires (76-, 25-, and 10-microm diameter), using microwave-assisted chemical vapor deposition (CVD). The diamond-coated wires were attached to copper wires (current collectors), and several methods were explored to insulate the cylindrical portion of the electrode: nail polish, epoxy, polyimide, and polypropylene coatings. The microelectrodes were characterized by scanning electron microscopy, Raman spectroscopy, and cyclic voltammetry. They exhibited low and stable background currents and sigmoidally shaped voltammetric curves for Ru(NH3)6(3+/2+) and Fe(CN)6(3-/4-) at low scan rates. The microelectrodes formed with the large diameter Pt and sealed in polypropylene pipet tips were employed for end-column detection in CE. Evaluation of the CE-EC system and the electrode performance were accomplished using a 10 mM phosphate buffer, pH 6.0, run buffer, and a 30-cm-long fused-silica capillary (75-microm i.d.) with dopamine, catechol, and ascorbic acid serving as test analytes. The background current (approximately 100 pA) and noise (approximately 3 pA) were measured at different detection potentials and found to be very stable with time. Reproducible separation (elution time) and detection (peak current or area) of dopamine, catechol, and ascorbic acid were observed with response precisions of 4.1% or less. Calibration curves constructed from the peak area were linear over 4 orders of magnitude, up to a concentration between 0.1 and 1 mM. Mass limits of detection for dopamine and catechol were 1.7 and 2.6 fmol, respectively (S/N = 3). The separation efficiency was approximately 33,000, 56,000, and 98,000 plates/m for dopamine, catechol, and ascorbic acid, respectively. In

  11. Quantification and speciation of technetium-99 in samples at low levels: contributions of capillary electrophoresis / ICP-MS system

    International Nuclear Information System (INIS)

    Given the low levels of 99Tc (long half-lived artificial radionuclide) in the environment (10-8 M to 10-12 M), its determination currently necessitates an enrichment and separation from the sample matrix prior to instrumental measurement. Therefore, nuclear safety monitoring requires the knowledge of the redox and chemical properties of this element in order to predict its behaviour and transfer in the environment. So, a separative and very sensitive measurement technique must thus be employed. We have developed a new environmental measurement method applied to the quantification and speciation of 99Tc in sample at environmental concentrations. Indeed, we have combined a Capillary Electrophoresis (CE) with an Inductively Coupled Plasma Mass Spectrometer (ICP-MS). The limit of detection of 99Tc is about 2.10-8 M by CE/ICP-MS system equipped with a PFA-50 nebuliser. In addition to the detection and measurement of technetium, we can separate online 99Tc(VII) of its interfering radionuclides like molybdenum and ruthenium by CE/ICP-MS. Indeed, due do the different migration time of each anions, it's possible to determinate a signal at m/z= 99 which is only given to 99Tc. Results obtained by this method have been compared to an usual radiochemical technique, extraction of Tc(VII) by a TEVA resin followed by ICP-MS measurement. Within the framework of storage of spent fuel, studies on the speciation of Tc(VII) by CE / ICP-MS iron-sulphide soils in anoxic conditions have shown that technetium VII is reduced by sulphured suspensions. (author)

  12. Interactions of non-charged tadalafil stereoisomers with cyclodextrins: capillary electrophoresis and nuclear magnetic resonance studies.

    Science.gov (United States)

    Fejős, Ida; Kazsoki, Adrienn; Sohajda, Tamás; Márványos, Ede; Volk, Balázs; Szente, Lajos; Béni, Szabolcs

    2014-10-10

    The single isomer drug R,R-tadalafil (Cialis) contains two chiral centers thus four stereoisomers (R,R-, S,S-, S,R- and R,S-tadalafil) exist, however, only the most potent inhibitor, the R,R-tadalafil is in clinical use. In our study, over 20 charged cyclodextrin (CD) derivatives were studied for enantiospecific host-guest type interactions in CD-modified capillary electrophoresis. Tadalafil stereoisomers are non-charged; therefore, their electrophoretic separation poses a challenge. Several candidates of both positively and negatively charged hosts were found to be effective for the enantioseparation. Eight out of the beta derivatives and three of alpha derivatives (including sulfated, sulfoalkylated, carboxyalkylated and amino derivatives) resolved all four stereoisomers partially or completely. Cavity size-dependent absolute enantiomer migration order (EMO) reversals were observed in the case of sulfopropyl-alpha (EMO: R,S; S,R; R,R; S,S) and sulfopropyl-beta (S,S; R,R; S,R; R,S) derivatives, while substituent-dependent partial EMO reversals were detected for sulfobutyl-ether-alpha (R,S; S,R; S,S; R,R) and sulfated-alpha-CD (R,R; S,S; R,S; S,R) selectors. Complexation-induced (1)H NMR chemical shift changes reflected that the benzodioxole moiety plays a major role in cavity size-dependent EMO reversal. Sulfobutyl-ether-alpha-CD was the only selector that provided the desired EMO in which the clinically applied eutomer R,R-tadalafil migrates last. Finally, an electrophoretic method applying a background electrolyte (BGE) containing 75 mM Tris-acetic acid buffer (pH 4.75) and 7 mM sulfobutyl-ether-alpha-CD was developed for the baseline resolution of all isomers at 25 °C and +25 kV applied voltage.

  13. Environmental water monitoring by capillary electrophoresis and result comparison with solvent chemistry techniques.

    Science.gov (United States)

    Sirén, Heli; Väntsi, Sirpa

    2002-05-24

    The aim of this work was to determine inorganic ions from natural waters by capillary electrophoresis (CE) and to compare the results obtained with those measured with conventional solvent chemistry techniques. The project was part of a larger CE study, during which we measured inorganic ions from some lake and river systems and groundwaters in Southern Finland. Results obtained from contaminated Finnish waters were compared with samples from the River Rhine in the Düsseldorf area. Two CE methods were used for analysis: one for determination of chloride, sulfate, nitrite and nitrate at pH 7.7 and the other for ammonium, potassium, calcium, sodium and magnesium at pH 3.6, both methods using identification based on indirect UV detection. Two separation methods were used in order to prevent complex formation of metals with sulfate, hydroxide and decomposed organic matter present in the environmental samples. On the basis of the CE studies dilution was needed for those samples having more than 100 mg/l of sulfate, chloride, calcium and sodium. On average, the natural waters in the study contained ammonium, magnesium, sodium, potassium and calcium below 0.3, 20, 200, 20, and 200 mg/l, respectively. The concentrations of chloride, sulfate, nitrite and nitrate were below 20, 100, 10, and 10 mg/l, respectively. Correlation of the CE results with those acquired by titration, atomic absorption spectrometry, ion chromatography and flow injection analysis were obtained; R2 values for the comparison tests varied from 0.8816 to 0.9994 depending on the ion. The repeatabilities of the anion and cation CE methods were tested using laboratory-made reference sample mixtures with high and low salt concentration. PMID:12102308

  14. Screening of Small-Molecule Inhibitors of Protein-Protein Interaction with Capillary Electrophoresis Frontal Analysis.

    Science.gov (United States)

    Xu, Mei; Liu, Chao; Zhou, Mi; Li, Qing; Wang, Renxiao; Kang, Jingwu

    2016-08-16

    A simple and effective method for identifying inhibitors of protein-protein interactions (PPIs) was developed by using capillary electrophoresis frontal analysis (CE-FA). Antiapoptotic B-cell-2 (Bcl-2) family member Bcl-XL protein, a 5-carboxyfluorescein labeled peptide truncated from the BH3 domain of Bid (F-Bid) as the ligand, and a known Bcl-XL-Bid interaction inhibitor ABT-263 were employed as an experimental model for the proof of concept. In CE-FA, the free ligand is separated from the protein and protein-ligand complex to permit the measurement of the equilibrium concentration of the ligand, hence the dissociation constant of the protein-ligand complex. In the presence of inhibitors, formation of the protein-ligand complex is hindered, thereby the inhibition can be easily identified by the raised plateau height of the ligand and the decayed plateau of the complex. Further, we proposed an equation used to convert the IC50 value into the inhibition constant Ki value, which is more useful than the former for comparison. In addition, the sample pooling strategy was employed to improve the screening throughput more than 10 times. A small chemical library composed of synthetic compounds and natural extracts were screened with the method, two natural products, namely, demethylzeylasteral and celastrol, were identified as new inhibitors to block the Bcl-XL-Bid interaction. Cell-based assay was performed to validate the activity of the identified compounds. The result demonstrated that CE-FA represents a straightforward and robust technique for screening of PPI inhibitors. PMID:27425825

  15. Polyvinylpyrrolidone-sodium dodecylsulfate complex is a family of pseudo-polyanions with different charge densities: Evidence from capillary electrophoresis, capillary viscosimetry and conductometry.

    Science.gov (United States)

    Wu, Yefan; Chen, Jie; Fang, Yun; Zhu, Meng

    2016-10-01

    Accordance with the previously supposed polyelectrolyte-like behaviour of neutral polymer-anionic surfactant complexes, direct evidence for the formation of the pseudo-polyanions in polyvinylpyrrolidone (PVP)-sodium dodecylsulfate (SDS) solution is put forward in this paper by capillary electrophoresis (CE) experiments in assistance with capillary viscosimetry and conductometry. The contradictory phenomena of the absolute value of relative electrophoretic mobility (re) increasing while the ionization degree (α) decreasing with the increasing specific clusterization [Г] in aqueous PVP-SDS solution are explained by the finding that the PVP-SDS complex is eventually a family of PVP-SDS pseudo-polyanions with different charge densities. And it is found countercations playing an important role in the formation of the PVP-SDS pseudo-polyanions in virtue of bridge effect. PMID:27348481

  16. Feasibility of applying the new improved PCR/LDR/capillary electrophoresis on prenatal diagnosis of fetal beta thalassaemia

    Directory of Open Access Journals (Sweden)

    Xi LIAO

    2014-03-01

    Full Text Available Objective  To investigate the feasibility of applying new improved polymerase chain reaction (PCR/ligase detection reaction (LDR/capillary electrophoresis to the prenatal diagnosis of fetal β thalassemia from maternal plasma. Methods  The mixture of different concentrations of normal DNA and trace β-thalassemia mutation heterozygous DNA were used for PCR, and the amplification products obtained thereafter were used for LDR. The ligation product was detected by capillary electrophoresis to demonstrate the sensitivity. Results  The amplification products containing same concentration of CD17 (A→T mutation were used as LDR template, 8U DNA ligase reaction was performed. LDR products were analyzed by genetic analyzer CEQ8000 type electrophoresis with a detection sensitivity of 1:5000. Product peak area decreased with the increase of normal peripheral blood DNA concentration gradient. When the concentration reached to 1:10 000, it would be hard to distinguish the product peak and miscellaneous peak. The LDR product was not detected in the product of negative control without CD17 (A→T mutation. The LDR results of normal maternal plasma DNA added 20pg CD17 (A→T heterozygous mutation in peripheral blood DNA amplification products revealed that the product peaks could be clearly distinguished from miscellaneous peaks. Conclusion  The new improved PCR/LDR/capillary electrophoresis technology is expected to be used in plasma DNA tests to prenatal diagnosis of fetal beta thalassaemia in pregnant women. DOI: 10.11855/j.issn.0577-7402.2014.03.06

  17. A New Method for Decreasing Mass Limit of Detection and Increasing Number of Theoretical Plates in Capillary Electrophoresis with Amperometric Detection

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    A new method was developed to decrease the mass limit of detection (LOD) and increase the number of theoretical plates (N) in capillary electrophoresis with amperometric detection. When the single microcylinder electrode, the 10 μm ID capillary with the etched detection end and the in-capillary alignment were used, the mass LOD for phenol was reduced 124 times and N was increased 36 times in comparison with the normal situation.

  18. Simultaneous determination of saccharin and aspartame in commercial noncaloric sweeteners using the PLS-2 multivariate calibration method and validation by capillary electrophoresis.

    Science.gov (United States)

    Cantarelli, Miguel A; Pellerano, Roberto G; Marchevsky, Eduardo J; Camiña, José M

    2008-10-22

    A new method to determine a mixture for sweetener sodium saccharin and aspartame in commercial noncaloric sweeteners is proposed. A classical full factorial design for standards was used in the calibration step to build the partial least-squares (PLS-2) model. Instrumental data were obtained by means of UV-visible spectrophotometry. Salicylic acid was used as an internal standard to evaluate the adjustment of the real samples to the PLS model. The concentration of analytes in the commercial samples was evaluated using the obtained model by UV spectral data. The PLS-2 method was validated by capillary zone electrophoresis (CZE), finding in all cases a relative error of less than 11% between the PLS-2 and the CZE methods. The proposed procedure was applied successfully to the determination of saccharin and aspartame in noncaloric commercial sweeteners.

  19. Supported liquid membrane extraction coupled in-line to commercial capillary electrophoresis for rapid determination of formate in undiluted blood samples.

    Science.gov (United States)

    Pantůčková, Pavla; Kubáň, Pavel; Boček, Petr

    2013-07-19

    A cheap, disposable sample pretreatment device with planar supported liquid membrane (SLM) was proposed, assembled and placed into an autosampler carousel of a commercial capillary electrophoresis (CE) instrument for automated pretreatment and analysis of formate in undiluted whole blood and serum samples. All analytical procedures except for filling the pretreatment device with donor and acceptor solutions, i.e., extraction across SLM, injection of the extracted sample and CE-UV determination of formate, were performed fully automatically. The pretreatment device required only μL volumes of blood sample and organic solvent per extraction and was disposed off after each extraction. Good repeatability of peak areas (≤7.7%) and migration times (≤1.5%), linear relationship (r(2)=0.998-0.999) and limits of detection (≤35μM) were achieved. The overall analytical process including blood withdrawal, filling the SLM device with respective solutions, extraction of blood sample, injection into separation capillary and CE separation of formate from other anions took less than 4min. The method was proved useful by direct determination of elevated formate concentrations in undiluted serum samples of a methanol intoxicated patient. Due to its compatibility with currently commercially available CE instrumentation, disposability of extraction devices, minimum sample handling/consumption, and short extraction/analysis times, the developed method might be attractive for rapid diagnosis of methanol poisoning in clinical and toxicological laboratories. PMID:23777836

  20. Elimination of the artefact peaks in capillary electrophoresis determination of glutamate by using organic solvents in sample preparation.

    Science.gov (United States)

    Campos, Camila Dalben Madeira; de Campos Braga, Patricia Aparecida; Reyes, Felix Guillermo Reyes; da Silva, José Alberto Fracassi

    2015-11-01

    Focusing on the demand from the food industry for fast and reliable alternative methods to control the quality of food products, we present in this paper a method for amino acid separation and glutamic acid quantification in complex matrices employing capillary electrophoresis with capacitively coupled contactless conductivity detection. We demonstrate by simulation and experimentally the use of organic solvents in sample preparation to prevent peak splitting and increase stacking in capillary electrophoretic separations of amino acids. Additionally, we obtained results for glutamic acid quantification comparable to those obtained via traditional methods used at industrial sites. We tested premium and low-cost samples with large variations in their glutamic acid content, which demonstrated the wide range of applicability of the method presented herein. The results of the proposed capacitively coupled contactless conductivity detection based capillary electrophoresis method agreed with those obtained by an enzymatic detector and ultra high performance liquid chromatography coupled to tandem mass spectrometry, considering a confidence level of 95%. PMID:26332708

  1. Capillary Zone Electrophoresis with Amperometric Detection for Composition Analysis of Laminarin

    Institute of Scientific and Technical Information of China (English)

    王清江; 丁飞; 李辉; 何品刚; 方禹之

    2003-01-01

    The composition of laminartn was firstly determined by analyz-ing its hydrolysis monosaccharides with capillary zone elec.lected opamum conditions, fucose, galactose, glucose, man-nose and xylose, which are hydrolysis products of iaminarin,could be perfectly separated within 20 min and showed signifi-cant current responses at copper electrodes. The linear ranges of fucose, galactose and glucose were from 1.0 × 10-6 to 2.0 ×10-4 mol·L-1, those of mannose and xylose were from 1.0×10-6 to 2.0× 10-4 mol·L-1, and their detect/on limits were at 10-7mol·L-1 level (S/N =3). The molar ratio of fucose,galactose,glucose,mannose and xylose in laminartn was 10.5:2.8:1.0:7.3:3.4 and the purity of this polysaccharide leached by the introduced leaching method was 95.7%. Compared to usual UV-vis and other spectrometric methods, analyzing polysaccharide by this method has some merit sof quickness,low-volume sampling,simple instrumentation,high sensitivity and high reproducibility.

  2. Capillary electrophoresis-mass spectrometry determination of morphine and its isobaric glucuronide metabolites.

    Science.gov (United States)

    Isbell, Theresa A; Strickland, Erin C; Hitchcock, Jennifer; McIntire, Gregory; Colyer, Christa L

    2015-02-01

    The determination of morphine and its isobaric metabolites morphine-3-beta-d-glucuronide (M3G) and morphine-6-beta-d-glucuronide (M6G) is useful for therapeutic drug monitoring and forensic identification of drug use. In particular, capillary electrophoresis with mass spectrometry (CE-MS) represents an attractive tool for opioid analysis. Whereas volatile background electrolytes in CE often improve electrospray ionization for coupled MS detection, such electrolytes may reduce CE separation efficiency and resolution. To better understand the effects of background electrolyte (BGE) composition on separation efficiency and detection sensitivity, this work compares and contrasts method development for both volatile (ammonium formate and acetate) and nonvolatile (ammonium phosphate and borate) buffers. Peak efficiencies and migration times for morphine and morphine metabolites were optimal with a 25mM ammonium borate buffer (pH=9.5) although greater sensitivities were achieved in the ammonium formate buffer. Optimized CE methods allowed for the resolution of the isobaric morphine metabolites prior to high mass accuracy, electrospray ionization quadrupole time-of-flight (ESI-QTOF) MS detection applicable to the analysis of urine samples in under seven minutes. Urine sample preparation required only a 10-fold dilution with BGE prior to analysis. Limits of detection (LOD) in normal human urine were found to be 1.0μg/mL for morphine and 2.5μg/mL for each of M3G and M6G by CE-ESI-QTOF-MS. These LODs were comparable to those for CE-UV analysis of opioid standards in buffer, whereas CE-ESI-QTOF-MS analysis of opioid standards in buffer yielded LODs an order of magnitude lower. Patient urine samples (N=12) were analyzed by this new CE-ESI-QTOF-MS method and no significant difference in total morphine content relative to prior liquid chromatography-mass spectrometry (LC-MS) results was found as per a paired-t test at the 99% confidence level. Whereas the LC-MS method applied

  3. Developments in and applications of capillary electrophoresis inductively coupled plasma mass spectrometry

    International Nuclear Information System (INIS)

    This project has set out to design and optimise a robust and efficient interface for capillary electrophoresis-inductively coupled plasma mass spectrometry (CE-ICP-MS) and to investigate the application of the technique in elemental speciation studies. An interface was constructed using a commercial microconcentric nebuliser (MCN) and a cyclonic spray chamber. The cyclonic spray chamber was designed specifically to provide rapid sample response and washout and to minimise sample dispersion. Isoforms of the heavy metal binding protein, metallothionein, were separated and the bound metals detected to characterise the interface. Suction from the self-aspirating nebuliser was identified as the principal factor controlling electrophoretic resolution. To maintain resolution, two methods for counterbalancing the nebuliser suction were investigated. In the first method an optimised make-up flow was employed, and in the second a negative pressure was applied to the buffer vial during the separation. The negative pressure method was preferred because it did not significantly compromise sensitivity. The MCN was found to be prone to regular blocking which compromised the analytical precision of the system. A second interface was constructed using a glass MicroMist nebuliser. The MicroMist nebuliser was found to be less prone to blocking than the MCN and significantly improved the precision of the system to less than 4.3% RSD. The MicroMist nebuliser did, however, provide a lower sensitivity. The advantage of employing an electroosmotic flow marker to correct for migration time drifts was demonstrated. A CE-ICP-MS method was developed for the speciation of selenium in selenium enriched yeasts and nutritional supplements. Selenoamino acids and inorganic selenium species were separated, as anions, under strong electroosmotic flow conditions. Methods to enhance the selenium sensitivity were investigated. A proteolytic enzyme extraction method was employed and the effect of the

  4. Characterization and stability of gold nanoparticles depending on their surface chemistry: Contribution of capillary zone electrophoresis to a quality control.

    Science.gov (United States)

    Pallotta, Arnaud; Boudier, Ariane; Leroy, Pierre; Clarot, Igor

    2016-08-26

    Four kinds of gold nanoparticles (AuNP) quite similar in terms of gold core size (ca. 5nm) and shape (spherical) but differing by their surface chemistry (either negatively, or positively charged, or neutral) were synthesized. They were analyzed using both the classical physicochemical approach (spectrophotometry, dynamic light scattering coupled or not to electrophoresis and transmission electron microscopy) and capillary zone electrophoresis equipped with photodiode array detection. The results obtained by both methodologies (related to Surface Plasmon Band-maximal absorbance wavelength-, and zeta potential and electrophoretic mobilities) were well correlated. Moreover, taking advantage of the separation method, the sample heterogeneity was evaluated and an impurity profile was extracted. This allowed setting some specifications which were then applied on the one hand to a batch-to-batch survey to declare NP as conform or not after production and on the other hand to a stability study. PMID:27435685

  5. Optimization strategies for separation of sulfadiazines using Box-Behnken design by liquid chromatography and capillary electrophoresis

    Institute of Scientific and Technical Information of China (English)

    GONG Wen-jun; ZHANG Yu-ping; ZHANG Yi-Jun; XU Guang-ri; WEI Xin-jun; LEE Kwang-pill

    2007-01-01

    Development of effective chromatographic or electrophoretic separation involves judicious deciding of selection of optimal experimental conditions that can provide an adequate resolution at a reasonable run time for the separation of interested components. Box-Behnken factorial design was effectively applied for the separation optimization of eight structurally related sulfonamides using capillary zone electrophorosis and reverse high performance liquid chromatography. Optimum values for volume ratio of THF to H2O in eluent, column temperature and flow rate of eluent are found as 12 to 88, 35 ℃ and 1.0 mL/min, respectively.Box-Behnken modified optimization model is extended to separation by capillary electrophoresis (CE). While using CE, a satisfactory separation is achieved with a minimum resolution larger than 1.0 for a separation time less than 10 min.

  6. Simultaneous determination of berberine,matrine and oxymatrine in traditional Chinese medicines by using nonaqueous capillary electrophoresis

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    A rapid method for the simultaneous determination of berberine(BBR),matrine(MT)and oxymatrine(OMT)by nonaqueous capillary electrophoresis(NACE)was developed.Optimum separation of the analytes was obtained on a 50cm×50μm i.d.fused-silica capillary using a non-aqueous buffer system of 70mM ammonium acetate,7.0% acetic acid and 10% acetonitrile at 25kV and 20℃.The relative standard deviations(R.S.D.)of the migration times and peak areas of the three active components were 0.06%-0.20% and 0.12%-3.41% for berber...

  7. Investigation of the in vitro biotransformation and simultaneous enantioselective separation of thalidomide and its neutral metabolites by capillary electrophoresis.

    Science.gov (United States)

    Weinz, C; Blaschke, G

    1995-12-15

    Reversed-phase liquid chromatography is a long established method for the analysis of drug metabolism. The current investigation demonstrates that micellar electrokinetic capillary chromatography can be an attractive alternative. Two methods were developed using sodium dodecyl sulfate and hexadecyltrimethylammonium bromide for the determination of possible hydroxylated metabolites of the former sedative drug thalidomide (Contergan) in order to study the in vitro metabolism of the drug by incubation with rat liver microsomes. The biotransformation was found to be stereoselective: S-(-)-thalidomide mainly formed 5-hydroxythalidomide, whereas R-(+)-thalidomide was preferentially transformed to two metabolites tentatively assigned to be diastereomers of 5'-hydroxythalidomide. Furthermore, the simultaneous enantioseparation of thalidomide and two of its possible hydroxylated metabolites was achieved using capillary electrophoresis with negatively charged carboxymethyl-beta-cyclodextrin. The dependencies of the selectivity of the enantioseparation on the concentration of the chiral additive and the pH of the run buffer were investigated. PMID:8788158

  8. Structural and conformational variants of human beta2-microglobulin characterized by capillary electrophoresis and complementary separation methods

    DEFF Research Database (Denmark)

    Heegaard, Niels H H; Rovatti, Luca; Nissen, Mogens H;

    2003-01-01

    of capillary temperature, organic solvent concentration, and analysis time. The results suggest that the apparent beta2-microglobulin heterogeneity observed by CE is caused by two distinct protein conformations that are present in beta2-microglobulin under partly denaturing conditions and that Met99-oxidized......The small (Mr = 11729) serum protein beta2-microglobulin is prone to precipitate as amyloid in a protein conformational disorder (PCD) that occurs in a significant number of patients on chronic hemodialysis. Analyses by capillary electrophoresis (CE) were undertaken to study beta2-microglobulin...... and normal (i.e. nonoxidized) beta2-microglobulin behave similarly with respect to the potential to attain this alternative conformation. CE is an attractive method to study early and intermediate soluble folding variants that may be involved in PCDs and CE thus may have an important role as a tool...

  9. Determination of carbohydrates in honey and milk by capillary electrophoresis in combination with graphene-cobalt microsphere hybrid paste electrodes.

    Science.gov (United States)

    Liang, Peipei; Sun, Motao; He, Peimin; Zhang, Luyan; Chen, Gang

    2016-01-01

    A graphene-cobalt microsphere (CoMS) hybrid paste electrode was developed for the determination of carbohydrates in honey and milk in combination with capillary electrophoresis (CE). The performance of the electrodes was demonstrated by detecting mannitol, sucrose, lactose, glucose, and fructose after CE separation. The five analytes were well separated within 9 min in a 40 cm long capillary at a separation voltage of 12 kV. The electrodes exhibited pronounced electrocatalytic activity, lower detection potentials, enhanced signal-to-noise characteristics, and higher reproducibility. The relation between peak current and analyte concentration was linear over about three orders of magnitude. The proposed method had been employed to determine lactose in bovine milk and glucose and fructose in honey with satisfactory results. Because only electroactive substances in the samples could be detected on the paste electrode, the electropherograms of both food samples were simplified to some extent.

  10. Development of chiral methodologies by capillary electrophoresis with ultraviolet and mass spectrometry detection for duloxetine analysis in pharmaceutical formulations.

    Science.gov (United States)

    Sánchez-López, Elena; Montealegre, Cristina; Marina, María Luisa; Crego, Antonio L

    2014-10-10

    Two chiral methodologies were developed by capillary electrophoresis (CE) with UV and mass spectrometry (MS) detection to ensure the quality control of the drug duloxetine, commercialized as a pure enantiomer. Both methods were optimized to achieve a high baseline enantioresolution (Rs>2) and an acceptable precision (RSD values developed methods were validated and applied for the first time to the analysis of four pharmaceutical formulations. The content of R-duloxetine in all these samples was below the detection limit and the amount of S-duloxetine was in good agreement with the labeled content, obtaining results by the two methods that did not differ significantly (p-values >0.05).

  11. A comparative study of capillary zone electrophoresis and pH-potentiometry for determination of dissociation constants.

    Science.gov (United States)

    Andrasi, Melinda; Buglyo, Peter; Zekany, Laszlo; Gaspar, Attila

    2007-09-01

    Acidity constants of six cephalosporin antibiotics, cefalexin, cefaclor, cefadroxil, cefotaxim, cefoperazon and cefoxitin are determined using capillary zone electrophoresis (CZE) and pH-potentiometric titrations. Since CZE is a separation method, it is not necessary for the samples to be of high purity and known concentration because only mobilities are measured. The effect on determination of dissociation constants of different matrices (serum, 0.9% NaCl, fermentation matrix) was examined. The advantages of CZE can be utilized in those fields where potentiometry has limitations (sample quantity, solubility, purity, simultaneous determinations), although pK(a) values that are close to each other can be determined by potentiometry with more accuracy.

  12. Using capillary electrophoresis mobility shift assay to study the interaction of CdTe quantum dots with bovine serum albumin

    Institute of Scientific and Technical Information of China (English)

    Li Wen Shao; Chao Qing Dong; Xiang Yi Huang; Ji Cun Ren

    2008-01-01

    In this work, the capillary electrophoresis mobility shift assay (CEMSA) was first adopted to study the interaction of protein with quantum dots (QDs). In this study, bovine serum albumin (BSA) and CdTe QDs were used as model samples. We observed that BSA was facilely adsorbed to CdTe QDs surface, and the QD-BSA complex was formed by a 1:1 stoichiometric ratio. A value of 2.17±0.27×106mol-1 L--1 (at 25℃) for the association constant was obtained by CEMSA.

  13. Precolumn affinity capillary electrophoresis for the identification of clinically relevant proteins in human serum: application to human cardiac troponin I.

    Science.gov (United States)

    Dalluge, J J; Sander, L C

    1998-12-15

    An approach has been developed to the on-line extraction and identification of clinical disease-state marker proteins in human serum. Fabrication of capillaries with integral packed beds for the online determination of human cardiac troponin I (cTnI), a diagnostic marker for myocardial infarction, at clinically relevant levels (2 nmol/L) in serum is demonstrated. The technique, termed precolumn affinity capillary electrophoresis (PA-CE), utilizes a short (approximately 5 mm) packed bed of porous silica containing covalently immobilized monoclonal anti-cTnI antibodies directly integrated within a separation capillary for the selective retention of cTnI from a complex matrix. Following a rinsing step to eliminate nonspecifically bound serum proteins and other impurities from the column, desorption of the antigen into the separation region of the PA-CE capillary for subsequent measurement of femto-molar amounts of cTnI by CE is effected by the injection of an appropriate elution buffer. Advantages of this approach over previously reported affinity preconcentration techniques, related applications for PA-CE technology, and its potential for use in the development of a certified reference material for cTnI in serum are discussed. PMID:9868922

  14. Diagnosis of a rare double heterozygous Hb D Punjab/Hb Q India hemoglobinopathy using Sebia capillary zone electrophoresis

    Directory of Open Access Journals (Sweden)

    Sushama Parab

    2014-01-01

    Full Text Available In India, hemoglobinopathies constitute a major genetic disorder and hemoglobin variants such as Hb S, Hb D Punjab, and Hb E are the most common ones. Other variants include Hb Q India, Hb Lepore, Hb J Meerut, Hb D Iran, etc. These variants show heterozygous state along with beta thalassemia. However, compound heterozygosities among these variants are very rare. Ethylenediaminetetraacetic acid whole blood sample received for routine thalassemia screening was subjected to alkaline electrophoresis using automated capillary zone electrophoresis. Suspecting the presence of rare variants, further analysis was carried out using Bio-Rad D10 and Tosoh G8 high-performance liquid chromatography (HPLC systems. Capillary zone electrophoretograms showed the presence of peaks in zone Hb A, Hb D, a fused peak in Hb A2, and a small peak in Z1 zone. Bio-Rad and Tosoh chromatograms also indicated the presence of four peaks which are identified as Hb A, Hb D Punjab, Hb Q India, and hybrid of Hb D Punjab/Hb Q India. A peak in Hb D zone of capillary was due to co-migration of Hb D Punjab and Hb Q India variants. Small peak in Z1 zone indicated the presence of alpha chain variant Hb Q India. The findings were further confirmed by HPLC results and molecular genetic studies. The present study reports for the 1 st time a rare hemoglobinopathy of double heterozygosity for Hb D Punjab, Hb Q India on Capillarys 2 Flex Piercing analyzer and is forth reported case for this rare hemoglobinopathy.

  15. Coated capillaries with highly charged polyelectrolytes and carbon nanotubes co-aggregated with sodium dodecyl sulphate for the analysis of sulfonylureas by capillary electrophoresis.

    Science.gov (United States)

    El-Debs, Racha; Nehmé, Reine; Claude, Bérengère; Motteau, Solène; Togola, Anne; Berho, Catherine; Morin, Philippe

    2014-11-01

    Sulfonylureas (SUs) are one of the most widely used herbicides to control weeds in crops. Herein, capillary electrophoresis (CE) was used to determine four sulfonylureas in natural waters, namely chlorsulfuron (CS), iodosulfuron methyl (IM), metsulfuron methyl (MSM) and mesosulfuron methyl (MSS). First of all, a bare silica capillary was chosen with 10mM of 1-butyl-3-methylimidazolium tetrafluoroborate (bminBF4) as electrophoretic buffer (pH 9.6) containing 2 mg L(-1) of surfactant-coated single-wall carbon nanotubes (SC-SWCNTs). A dramatic deviation in migration times was observed. Therefore, a poly(diallyldimethylammonium) chloride (PDADMAC) statically coated cationic capillary was used to improve repeatability and to alter the selectivity of the separation. The electroosmotic flow (EOF) measurement revealed that the SC-SWCNTs were strongly adsorbed at the surface of the PDADMAC coating even in the absence of the surfactant-coated nanotubes in the electrolyte buffer. Consequently, a stable strong cathodic EOF and excellent repeatabilities were obtained with relative standard deviations (RSDs) on migration times and on corrected peak areas below 0.9 and 1.5%, respectively. The separation of the SUs was conducted in only 6 min. No regeneration of the coating between analyses was necessary, and high peak efficiencies up to 173,000 theoretical plates were obtained. The bi-layer coating was subsequently used to analyze sulfonylureas in tap water, in several mineral waters as well as in underground waters spiked with SUs and directly injected into the CE capillary.

  16. New multilayer coating using quaternary ammonium chitosan and κ-carrageenan in capillary electrophoresis: application in fast analysis of betaine and methionine.

    Science.gov (United States)

    Vitali, Luciano; Della Betta, Fabiana; Costa, Ana Carolina O; Vaz, Fernando Antonio Simas; Oliveira, Marcone Augusto Leal; Vistuba, Jacqueline Pereira; Fávere, Valfredo T; Micke, Gustavo A

    2014-06-01

    The aim of this study was to develop a new multilayer coating with crosslinked quaternary ammonium chitosan (hydroxypropyltrimethyl ammonium chloride chitosan; HACC) and κ-carrageenan for use in capillary electrophoresis. A new semi-permanent multilayer coating was formed using the procedure developed and the method does not require the presence of polymers in the background electrolyte (BGE). The new capillary multilayer coating showed a cathodic electroosmotic flow (EOF) of around 30×10(-9) m(2) V(-1) s(-1) which is pH-independent in the range of pH 2 to 10. The enhanced EOF at low pH obtained contributed significantly to the development of a fast method of separation. The multilayer coating was then applied in the development of a fast separation method to determine betaine and methionine in pharmaceutical formulations by capillary zone electrophoresis (CZE). The BGE used to determine the betaine and methionine concentrations was composed of 10 mmol L(-1) tris(hydroxymethyl) aminomethane, 40 mmol L(-1) phosphoric acid and 10% (v/v) ethanol, at pH 2.1. A fused-silica capillary of 32 cm (50 µm ID×375 µm OD) was used in the experiments and samples and standards were analyzed employing the short-end injection procedure (8.5 cm effective length). The instrumental analysis time of the optimized method was 1.53 min (approx. 39 runs per hour). The validation of the proposed method for the determination of betaine and methionine showed good linearity (R(2)>0.999), adequate limit of detection (LOD <8 mg L(-1)) for the concentration in the samples and inter-day precision values lower than 3.5% (peak area and time migration). The results for the quantification of the amino acids in the samples determined by the CZE-UV method developed were statistically equal to those obtained with the comparative LC-MS/MS method according to the paired t-test with a confidence level of 95%. PMID:24725863

  17. Separation of poly(acrylic acid) salts according to topology using capillary electrophoresis in the critical conditions.

    Science.gov (United States)

    Maniego, Alison R; Ang, Dale; Guillaneuf, Yohann; Lefay, Catherine; Gigmes, Didier; Aldrich-Wright, Janice R; Gaborieau, Marianne; Castignolles, Patrice

    2013-11-01

    Branching was detected in polyacrylates synthesised through radical polymerization via solution-state NMR, while inconsistencies have been reported for the determination of the molar mass of hydrophilic polyacrylates using aqueous-phase and organic-phase size-exclusion chromatography. In this work, poly(sodium acrylate)s, PNaAs, of various topologies were separated for the first time using free-solution capillary electrophoresis (CE). Free-solution CE does not separate the PNaAs by their molar mass, similarly to separations by liquid chromatography in the critical conditions, rather by different topologies (linear, star branched, and hyperbranched). The electrophoretic mobility of PNaAs increases as the degree of branching decreases. Separation is shown to be not only by the topology but also by the end groups as expected for a separation in the critical conditions: replacing a relatively bulky nitroxide end group with hydrogen atom yielded a higher electrophoretic mobility. This novel method, capillary electrophoresis in the critical conditions enabled, for the first time, the separation of hydrophilic polyacrylates according to their topology (branching) and their chain ends. This will allow meaningful and accurate characterization of their branched topologies as well as molar masses and progress in for advanced applications such as drug delivery or flocculation. PMID:23732867

  18. The inclusion complex of rosmarinic acid into beta-cyclodextrin: A thermodynamic and structural analysis by NMR and capillary electrophoresis.

    Science.gov (United States)

    Aksamija, Amra; Polidori, Ange; Plasson, Raphaël; Dangles, Olivier; Tomao, Valérie

    2016-10-01

    This work focuses on the characterization of the rosmarinic acid (RA)-β-cyclodextrin (CD) complex in aqueous solution by (1)H NMR (1D- and 2D-ROESY), completed with studies by capillary electrophoresis (CE). From the (1)H NMR data, the stoichiometry of the complex was determined by a Job's plot and the binding constant was estimated from a linear regression (Scott's method). At pH 2.9, the results showed that RA binds CD with a 1:1 stoichiometry and a binding constant Kb of 445 (±53) M(-1) or 465 (±81) M(-1) depending on the CD protons (H-5 or H-3) selected for the evaluation. The Kb value was also calculated from the CD-induced chemical shifts of each RA proton in order to collect information on the structure of the complex. The pH dependence of Kb revealed that the RA carboxylic form displays the highest affinity for CD. An investigation by capillary electrophoresis fully confirmed these results. 2D ROESY analysis provided detailed structural information on the complex and showed a strong correlation between H-3 and H-5 of CD and most RA protons. In conclusion, RA, an efficient phenolic antioxidant from rosemary with a marketing authorization, spontaneously forms a relatively stable inclusion complex with CD in water. PMID:27132848

  19. On-line coupling of immobilized cytochrome P450 microreactor and capillary electrophoresis: A promising tool for drug development.

    Science.gov (United States)

    Schejbal, Jan; Řemínek, Roman; Zeman, Lukáš; Mádr, Aleš; Glatz, Zdeněk

    2016-03-11

    In this work, the combination of an immobilized enzyme microreactor (IMER) based on the clinically important isoform cytochrome P450 2C9 (CYP2C9) with capillary electrophoresis (CE) is presented. The CYP2C9 was attached to magnetic SiMAG-carboxyl microparticles using the carbodiimide method. The formation of an IMER in the inlet part of the separation capillary was ensured by two permanent magnets fixed in a cassette from the CE apparatus in the repulsive arrangement. The resulting on-line system provides an integration of enzyme reaction mixing and incubation, reaction products separation, detection and quantification into a single fully automated procedure with the possibility of repetitive use of the enzyme and minuscule amounts of reactant consumption. The on-line kinetic and inhibition studies of CYP2C9's reaction with diclofenac as a model substrate and sulfaphenazole as a model inhibitor were conducted in order to demonstrate its practical applicability. Values of the apparent Michalis-Menten constant, apparent maximum reaction velocity, Hill coefficient, apparent inhibition constant and half-maximal inhibition concentration were determined on the basis of the calculation of the effective substrate and inhibitor concentrations inside the capillary IMER using a model described by the Hagen-Poisseulle law and a novel enhanced model that reflects the influence of the reactants' diffusion during the injection process. PMID:26877175

  20. A capillary electrophoresis method to explore the self-assembly of a novel polypeptide ligand with quantum dots.

    Science.gov (United States)

    Wang, Jianhao; Zhang, Chencheng; Liu, Li; Kalesh, Karunakaran A; Qiu, Lin; Ding, Shumin; Fu, Minli; Gao, Li-Qian; Jiang, Pengju

    2016-08-01

    Polyhistidine peptides are effective ligands to coat quantum dots (QDs). It is known that both the number of histidine (His) residues repeats and their structural arrangements in a peptide ligand play important roles in the assembly of the peptide onto CdSe/ZnS QDs. However, due to steric hindrance, a peptide sequence with more than six His residue tandem repeats would hardly coordinate well with Zn(2+) in the QD shell to further enhance the binding affinity. To solve this problem, a His-containing peptide ligand, ATTO 590-E2 G (NH)6 (ATTO-NH), was specifically designed and synthesized for assembly with QDs. With sequential injection of QDs and ATTO-NH into the capillary electrophoresis with fluorescence detection, strong Förster resonance energy transfer phenomenon between the QDs and the ATTO 590 dye was observed, indicating efficient self-assembly of the novel peptide onto the QDs to form ATTO-NH capped QDs inside the capillary. The binding stability of the ligand onto the QD was then systematically investigated by titrating with imidazole, His, and a his-tag containing competitive peptide. It is believed that this new in-capillary assay significantly reduced the sample consumption and the analysis time. By functionalizing QDs with certain metal cation-specific group fused peptide ligand, the QD-based probes could be even extended to the online detection of metal cations for monitoring environment in the future. PMID:27334251

  1. Determination of arbutin and bergenin in Bergeniae Rhizoma by capillary electrophoresis with a carbon nanotube-epoxy composite electrode.

    Science.gov (United States)

    Zhang, Luyan; Zhang, Wei; Chen, Gang

    2015-11-10

    This report describes the fabrication and the application of a novel carbon nanotube (CNT)-epoxy composite electrode as a sensitive amperometric detector for the capillary electrophoresis (CE). The composite electrode was fabricated on the basis of the in situ polycondensation of a mixture of CNTs and 1,2-ethanediamine-containing bisphenol A epoxy resin in the inner bore of a piece of fused silica capillary under heat. It was coupled with CE for the separation and detection of arbutin and bergenin in Bergeniae Rhizoma, a traditional Chinese medicine, to demonstrate its feasibility and performance. The two phenolic constituents were well separated within 10min in a 45cm capillary length at a separation voltage of 12kV using a 50mM borate buffer (pH 9.2). The CNT-based detector offered higher sensitivity, significantly lower operating potential, satisfactory resistance to surface fouling, and lower expense of operation, indicating great promise for a wide range of analytical applications. It showed long-term stability and reproducibility with relative standard deviations of less than 5% for the peak current (n=15).

  2. Optimization of capillary electrophoresis method with contactless conductivity detection for the analysis of tobramycin and its related substances.

    Science.gov (United States)

    El-Attug, Mohamed Nouri; Hoogmartens, Jos; Adams, Erwin; Van Schepdael, Ann

    2012-01-25

    A method was validated and optimized to determine tobramycin (TOB) and its related substances. TOB is an aminoglycoside antibiotic which lacks a strong UV absorbing chromophore or fluorophore. Due to the physicochemical properties of TOB, capillary electrophoresis (CE) in combination with Capacitively Coupled Contactless Conductivity Detection (C(4)D) was chosen. The optimized separation method uses a background electrolyte (BGE) composed of 25 mM morpholinoethane-sulphonic acid (MES) adjusted to pH 6.4 by L-histidine (l-His). 0.3 mM cetyltrimethyl ammonium bromide (CTAB) was added as electroosmotic flow modifier in a concentration below the critical micellar concentration (CMC). Ammonium acetate 50 mg L(-1) was used as internal standard (IS). 30 kV was applied in reverse polarity (cathode at the injection capillary end) on a fused silica capillary (65/43 cm; 75 μm id). The optimized separation was obtained in less than 7 min with good linearity (R(2)=0.9995) for tobramycin. It shows a good precision expressed as RSD on relative peak areas equal to 0.2% and 0.7% for intraday and interday respectively. The LOD and LOQ are 0.4 and 1.3 mg L(-1) corresponding to 9 pg and 31 pg respectively. PMID:22015240

  3. Determination of arbutin and bergenin in Bergeniae Rhizoma by capillary electrophoresis with a carbon nanotube-epoxy composite electrode.

    Science.gov (United States)

    Zhang, Luyan; Zhang, Wei; Chen, Gang

    2015-11-10

    This report describes the fabrication and the application of a novel carbon nanotube (CNT)-epoxy composite electrode as a sensitive amperometric detector for the capillary electrophoresis (CE). The composite electrode was fabricated on the basis of the in situ polycondensation of a mixture of CNTs and 1,2-ethanediamine-containing bisphenol A epoxy resin in the inner bore of a piece of fused silica capillary under heat. It was coupled with CE for the separation and detection of arbutin and bergenin in Bergeniae Rhizoma, a traditional Chinese medicine, to demonstrate its feasibility and performance. The two phenolic constituents were well separated within 10min in a 45cm capillary length at a separation voltage of 12kV using a 50mM borate buffer (pH 9.2). The CNT-based detector offered higher sensitivity, significantly lower operating potential, satisfactory resistance to surface fouling, and lower expense of operation, indicating great promise for a wide range of analytical applications. It showed long-term stability and reproducibility with relative standard deviations of less than 5% for the peak current (n=15). PMID:26263060

  4. Simultaneous determination of caffeine, paracetamol, and ibuprofen in pharmaceutical formulations by high-performance liquid chromatography with UV detection and by capillary electrophoresis with conductivity detection.

    Science.gov (United States)

    Cunha, Rafael R; Chaves, Sandro C; Ribeiro, Michelle M A C; Torres, Lívia M F C; Muñoz, Rodrigo A A; Dos Santos, Wallans T P; Richter, Eduardo M

    2015-05-01

    Paracetamol, caffeine and ibuprofen are found in over-the-counter pharmaceutical formulations. In this work, we propose two new methods for simultaneous determination of paracetamol, caffeine and ibuprofen in pharmaceutical formulations. One method is based on high-performance liquid chromatography with diode-array detection and the other on capillary electrophoresis with capacitively coupled contactless conductivity detection. The separation by high-performance liquid chromatography with diode-array detection was achieved on a C18 column (250×4.6 mm(2), 5 μm) with a gradient mobile phase comprising 20-100% acetonitrile in 40 mmol L(-1) phosphate buffer pH 7.0. The separation by capillary electrophoresis with capacitively coupled contactless conductivity detection was achieved on a fused-silica capillary (40 cm length, 50 μm i.d.) using 10 mmol L(-1) 3,4-dimethoxycinnamate and 10 mmol L(-1) β-alanine with pH adjustment to 10.4 with lithium hydroxide as background electrolyte. The determination of all three pharmaceuticals was carried out in 9.6 min by liquid chromatography and in 2.2 min by capillary electrophoresis. Detection limits for caffeine, paracetamol and ibuprofen were 4.4, 0.7, and 3.4 μmol L(-1) by liquid chromatography and 39, 32, and 49 μmol L(-1) by capillary electrophoresis, respectively. Recovery values for spiked samples were between 92-107% for both proposed methods.

  5. On-line coupling of cyclodextrin mediated nonaqueous capillary electrophoresis to mass spectrometry for the determination of salbutamol enantiomers in urine

    OpenAIRE

    Servais, Anne-Catherine; Fillet, Marianne; Mol, Roelof; Somsen, G. W.; Chiap, Patrice; de Jong, G.J.; Crommen, Jacques

    2006-01-01

    The usefulness of the on-line coupling of nonaqueous capillary electrophoresis (NACE) with electrospray ionization (ESI) mass spectrometry (MS) using heptakis(2,3-di-O-acetyl-6-O-sulfo)-beta-cyclodextrin (HDAS-beta-CD) was demonstrated for the enantioselective determination of low concentrations of salbutamol in human urine. After optimization of several parameters, such as sheath-liquid composition and flow rate, nebulizing gas pressure, CE counter-pressure and position of the CE capillary o...

  6. Developments in coupled solid-phase extraction-capillary electrophoresis 2011-2013

    NARCIS (Netherlands)

    Ramautar, Rawi; Somsen, Govert W; de Jong, Gerhardus J

    2014-01-01

    This article presents an overview of the design and application of coupled SPE-CE systems that have been reported in the literature between January 2011 and June 2013. The present paper is an update of three previous review papers covering the years 2000-2011 (Electrophoresis 2008, 29, 108-128; Elec

  7. Study of the Electrophoretic Behavior of Cephalosporins by Capillary Zone Electrophoresis

    OpenAIRE

    Gabriel Hancu; Adina Sasebeşi; Aura Rusu; Hajnal Kelemen; Adriana Ciurba

    2015-01-01

    Purpose: The aim of the study was the characterization of the electrophoretic behavior of cephalosporins from different generation having different structural characteristics in order to develop a rapid, simple and efficient capillary electrophoretic method for their identification and simultaneous separation from complex mixtures. Methods: Ten cephalosporin derivatives (cefaclor, cefadroxil, cefalexin, cefazolin, cefoxitin, cefuroxime, cefoperazone, cefotaxime, ceftazidime, ce...

  8. Electrochemical Enzyme Immunoassay of Tumor Marker CA15-3 with Capillary Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Tumor marker CA15-3 was determined by using capillary electrophoretic enzyme immunoassay with electrochemical detection (CE-EIA-ED). The method can be used to detect CA15-3 with a limit of 0.024 U/mL.

  9. [The novel copolymer coated capillary columns of electrophoresis and their applications to separation of proteins].

    Science.gov (United States)

    Lu, G; Gao, D; Gu, J; Fu, R; Li, F; Zhang, H

    1999-01-01

    The copolymer of acrylonitrile, methyl acrylate, hydroxy ethyl acrylate (ZB-004), the copolymer of acrylonitrile, methyl acrylate, hydroxy ethyl acrylate, acrylamide (ZB-014) and the copolymer of acrylonitrile, hydroxy ethyl acrylate (ZB-016) were coated on the inner surface of fused-silica capillaries by just filling the capillary with solutions containing these copolymers followed by flushing the capillary with nitrogen. The physically adsorbed layer can reduce both protein adsorption and electroosmotic flow in the pH range of 3-5. Electroosmotic flow decreased by raising the concentrations of the copolymers. Separation performance of ZB-004 layer is better than those of other two layers due to its low hydrophilicity, but with higher pH values, appreciable peak deformation and increase in electroosmosis were observed. The intra day and inter day migration reproducibility were investigated in terms of relative standard deviation (RSD) with four basic proteins at pH 4.0. The RSDs of the intra day migration times were less than 2%. The RSDs of the inter day migration times were less than 4%. At pH 5.0, the RSDs of the migration times in two ZB-004-coated capillaries made on two different days were less than 1%. Separation efficiencies of four basic proteins in a ZB-004-coated capillary which stored in a buffer (pH 4.0) for fifteen days after being used for 14 days decreased 15%. These coatings were stable and exhibited reproducible separations from intra day, inter day and inter column under acidic conditions.

  10. Pneumatic Microvalve-Based Hydrodynamic Sample Injection for High-Throughput, Quantitative Zone Electrophoresis in Capillaries

    Energy Technology Data Exchange (ETDEWEB)

    Kelly, Ryan T.; Wang, Chenchen; Rausch, Sarah J.; Lee, Cheng S.; Tang, Keqi

    2014-07-01

    A hybrid microchip/capillary CE system was developed to allow unbiased and lossless sample loading and high throughput repeated injections. This new hybrid CE system consists of a polydimethylsiloxane (PDMS) microchip sample injector featuring a pneumatic microvalve that separates a sample introduction channel from a short sample loading channel and a fused silica capillary separation column that connects seamlessly to the sample loading channel. The sample introduction channel is pressurized such that when the pneumatic microvalve opens briefly, a variable-volume sample plug is introduced into the loading channel. A high voltage for CE separation is continuously applied across the loading channel and the fused silica capillary separation column. Analytes are rapidly separated in the fused silica capillary with high resolution. High sensitivity MS detection after CE separation is accomplished via a sheathless CE/ESI-MS interface. The performance evaluation of the complete CE/ESI-MS platform demonstrated that reproducible sample injection with well controlled sample plug volumes could be achieved by using the PDMS microchip injector. The absence of band broadening from microchip to capillary indicated a minimum dead volume at the junction. The capabilities of the new CE/ESI-MS platform in performing high throughput and quantitative sample analyses were demonstrated by the repeated sample injection without interrupting an ongoing separation and a good linear dependence of the total analyte ion abundance on the sample plug volume using a mixture of peptide standards. The separation efficiency of the new platform was also evaluated systematically at different sample injection times, flow rates and CE separation voltages.

  11. Isotachophoretic zone formation of serum albumin in different free fluid electrophoresis instruments.

    Science.gov (United States)

    Thormann, W; Firestone, M A; Sloan, J E; Long, T D; Mosher, R A

    1990-04-01

    The isotachophoretic behavior of a model protein, serum albumin, was examined (i) by computer simulation, (ii) by capillary isotachophoresis in HPE 100 and Tachophor 2127, (iii) by continuous flow isotachophoresis in Elphor VaP 22 and the BIO-STREAM Separator and (iv) by recycling isotachophoresis in an apparatus of our own design. Variations in monitored zone shapes can be explained by differences in engineering aspects and fluid stabilization principles of the instruments. PMID:2340824

  12. Capillary electrophoresis with laser-induced fluorescence detection for studying amino acid uptake by yeast during beer fermentation.

    Science.gov (United States)

    Turkia, Heidi; Sirén, Heli; Penttilä, Merja; Pitkänen, Juha-Pekka

    2015-01-01

    The amino acid composition of cultivation broth is known to affect the biomass accumulation, productivity, and vitality of yeast during cultivation. A separation method based on capillary electrophoresis with laser-induced fluorescence (LIF) detection was developed for the determination of amino acid consumption by Saccharomyces cerevisiae during beer fermentation. Intraday relative standard deviations were less than 2.1% for migration times and between 2.9% and 9.9% for peak areas. Interday relative standard deviations were less than 2.5% for migration times and between 4.4% and 18.9% for peak areas. The quantification limit was even as low as 62.5 pM which equals to below attomole level detection. The method was applied to study the rate of amino acid utilization during beer fermentation.

  13. A Compact System of Capacitively Coupled Contactless Conductivity Detection Based on the Square Wave Excitation Signal for Capillary Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    Shui Feng ZHANG; Li Shi WANG; Zhi DANG

    2006-01-01

    This paper reported a compact system of capacitively coupled contactless conductivity detection (C4D) based on the square wave excitation voltage for capillary electrophoresis, and it exhibited excellent sensitivity at the optimal frequency of 198 kHz. The feasibility and sensitivity of this detector was demonstrated by simultaneous detection of thirteen ions including alkali,alkaline earth and heavy metal ions. And the detection limits (S/N 3) were in the range of 0.2-1μmol/L for Mn2+,K+,Na+,Mg2+,Ca2+ ,Li+ ,Ba2+ ,and 7-25μmol/L for Ni2+ ,Cu2+ ,Cd2+ ,Pb2+,Co2+ ,Zn2+.

  14. Simultaneous determination of active ingredients in Erigeron breviscapus (Vant.) Hand-Mazz. by capillary electrophoresis with electrochemical detection.

    Science.gov (United States)

    Chu, Qingcui; Wu, Ting; Fu, Liang; Ye, Jiannong

    2005-03-01

    A high-performance capillary electrophoresis (CE) with electrochemical detection (ED) method was developed for the determination of the pharmacologically active ingredients in Erigeron breviscapus (Vant.) Hand-Mazz. and its extract phytopharmaceuticals in this work. Under the optimum conditions, nine analytes, baicalein, naringenin, scopoletin, kaempferol, apigenin, scutellarin, luteolin, caffeic acid and protocatechuic acid were separated within 24 min in a borax buffer (pH 8.7). Notably, excellent linearity was obtained over two orders of magnitude with detection limits (S/N=3) ranged from 1.0 x 10(-7) g/mL to 5.6 x 10(-7) g/mL for all nine analytes. This method was successfully used in the analysis of E. breviscapus (Vant.) Hand-Mazz. and its phytopharmaceuticals with a relatively simple extraction procedure, and the assay results were satisfactory. PMID:15740914

  15. Study of Interaction between Red-tide Toxin, Domoic Acid and Double -stranded DNA by Capillary Zone Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    Da Zhi LI; Xin Ya HE; Hui WANG; Li SUN; Bing Cheng LIN

    2004-01-01

    The interactions between amnesic red-tide toxin, domoic acid (DA) and 14mer double-stranded DNA (dsDNA with three kinds of sequences) were studied by capillary zone electrophoresis (CZE). For the dsDNA with a sequence of 5'-CCCCCTATACCCGC-3', the amount of free dsDNA decreases with the increase of added DA; and the signal of DA-dsDNA complex was observed. Meanwhile, the other two dsDNAs, 5'-(C)12GC-3' and 5'-(AT)7-3', the existence of DA could not lead to the change of dsDNA signal and indicated that there is no interaction between DA and these two dsDNAs.

  16. Migration behavior of alkylphenols, bisphenol A and bisphenol S studied by capillary electrophoresis using sulfated beta-cyclodextrin.

    Science.gov (United States)

    Mori, M; Naraoka, H; Tsue, H; Morozumi, T; Kaneta, T; Tanaka, S

    2001-06-01

    An application of capillary electrophoresis (CE) using sulfated beta-cyclodextrin (SCD) has been investigated for separating alkylphenols with different chain lengths, as well as bisphenol A and bisphenol S. In the absence of SCD in running buffer, all the phenols migrated at the same velocity as the electroosmotic flow (EOF), whereas the addition of SCD effectively led to the baseline separation of alkylphenols on the basis of the difference in the abilities to bind into the hydrophobic cavity of CD. The host-guest binding constants between analyte phenols and SCD were evaluated from Benesi-Hildebrand plots of the data obtained by two independent methods, CE and UV-visible measurements, demonstrating that the greater the hydrophobicity of the phenols, the larger the binding constants. The effects of organic solvents on the resolution for alkylphenols and bisphenols were also examined. This system using SCD was effective for the separation of 4-octylphenol and 4-nonylphenol isomers having longer alkyl chains.

  17. Simultaneous chromatographic fingerprinting and quantitative analysis of flavonoids in Pollen Typhae by high-performance capillary electrophoresis

    Directory of Open Access Journals (Sweden)

    Le Han

    2012-12-01

    Full Text Available To evaluate the quality of Pollen Typhae as used in traditional Chinese medicine, a high-performance capillary electrophoresis (HPCE method has been developed, validated and applied to chromatographic fingerprinting and quantitation of its eight main bioactive flavonoids (naringenin, isorhamnetin 3-O-(2G-α-l-rhamnosyl-rutinoside, rhamnetin 3-O-neohesperidoside, isorhamnetin, quercetin 3-O-(2G-α-l-rhamnosyl-rutinoside, quercetin 3-O-neohesperidoside, kaempferol and quercetin. Fingerprinting was based on the selection of nine characteristic chromatographic peaks. In quantitative analysis, the recovery of all eight compounds was in the range 98.5–102.2% with good linearity (r2>0.9919 over a relatively wide concentration range. The assay was successfully applied to the analysis of the eight bioactive flavonoids in 14 different samples. The results indicate that the assay is reproducible and precise and can be used for convenient quality assessment of Pollen Typhae.

  18. Determination of Aniline and Its Derivatives in Environmental Water by Capillary Electrophoresis with On-Line Concentration

    Directory of Open Access Journals (Sweden)

    Jianzhi Sun

    2012-06-01

    Full Text Available This paper describes a simple, sensitive and environmentally benign method for the direct determination of aniline and its derivatives in environmental water samples by capillary zone electrophoresis (CZE with field-enhanced sample injection. The parameters that influenced the enhancement and separation efficiencies were investigated. Surprisingly, under the optimized conditions, two linear ranges for the calibration plot, 1–50 ng/mL and 50–1000 ng/mL (R > 0.998, were obtained. The detection limit was in the range of 0.29–0.43 ng/mL. To eliminate the effect of the real sample matrix on the stacking efficiency, the standard addition method was applied to the analysis of water samples from local rivers.

  19. Monitoring the ionic content of exhaled breath condensate in various respiratory diseases by capillary electrophoresis with contactless conductivity detection.

    Science.gov (United States)

    Greguš, Michal; Foret, František; Kindlová, Dagmar; Pokojová, Eva; Plutinský, Marek; Doubková, Martina; Merta, Zdeněk; Binková, Ilona; Skřičková, Jana; Kubáň, Petr

    2015-06-01

    The analysis of an ionic profile of exhaled breath condensate (EBC) by capillary electrophoresis with contactless conductivity detection and double opposite end injection, is demonstrated. A miniature sampler made from a 2 ml syringe and an aluminium cooling cylinder was used for the fast collection of EBC (under one minute). Analysis of the collected EBC was performed in a 60 mM 2-(N-morpholino)ethanesulfonic acid, 60 mM L-histidine background electrolyte with 30 µM cetyltrimethylammonium bromide and 2 mM 18-crown-6 at pH 6, and excellent repeatability of migration times (RSD  acetate, lactate). The developed collection system and method provides a highly reproducible and fast way of collecting and analyzing EBC, with future applicability in point-of-care diagnostics. PMID:25944821

  20. Authentication of coffee by means of PCR-RFLP analysis and lab-on-a-chip capillary electrophoresis.

    Science.gov (United States)

    Spaniolas, Stelios; May, Sean T; Bennett, Malcolm J; Tucker, Gregory A

    2006-10-01

    Coffee is one of the most important world food commodities, commercial trade consisting almost entirely of Arabica and Robusta varieties. The former is considered to be of superior quality and thus attracts a premium price. Methods to differentiate these coffee species could prove to be beneficial for the detection of either deliberate or accidental adulteration. This study describes a molecular genetics approach to differentiate Arabica and Robusta coffee beans. This employs a Polymerase Chain Reaction-Restriction Fragment Length Polymorphism to monitor a single nucleotide polymorphism within the chloroplastic genome. Samples were analyzed with a lab-on-a-chip capillary electrophoresis system. Coffee powder mixtures were analyzed with this technique, displaying a 5% limit of detection. The plastid copy number was found to be relatively constant across a wide range of bean samples, suggesting that this methodology can also be employed for the quantification of any adulteration of Arabica with Robusta beans. PMID:17002409

  1. Simultaneous determination of electroactive and non-electroactive food preservatives by novel capillary electrophoresis with amperometric detection.

    Science.gov (United States)

    Wang, Weiyu; Wang, Yiping; Zhang, Junbo; Chu, Qingcui; Ye, Jiannong

    2010-09-23

    A novel capillary electrophoresis and amperometric detection method was achieved by adding an electroactive additive (3,4-dihydroxybenzylamine, 3,4-DHBA) to the running buffer, so that both electroactive and non-electroactive food preservatives were simultaneously determined. Under the selected optimum conditions, four electroactive preservatives (methylparaben, ethylparaben, propylparaben and butylparaben) and two non-electroactive preservatives (potassium sorbate and sodium lactate) were well separated and sensitively detected with detection limits (S/N=3) ranging from 1.06×10(-8) to 2.73×10(-6) g mL(-1). This method has been successfully employed for the determination of both electroactive and non-electroactive preservatives in several food commodities.

  2. Simultaneous analysis of acidulants and preservatives in food samples by using capillary zone electrophoresis with indirect UV detection.

    Science.gov (United States)

    Yoshikawa, Kenji; Saito, Shintaro; Sakuragawa, Akio

    2011-08-01

    Capillary zone electrophoresis with indirect UV detection was developed for the simultaneous analysis of acidulants and preservatives in food samples. When a solution of tris (hydroxymethyl) aminomethane, trimellitic acid and poly (vinyl alcohol) was used as the background electrolyte, the nine acidulants and four preservatives listed in the Japanese Food Sanitation Law were detected within 8min. The calibration curves plotted from the peak height of each analyte were linear with a correlation coefficient of 0.99. The relative standard deviations (n=10) of the peak height ranged from 1.2% to 4.7%. The detection limits for these species ranged from 0.6 to 5.3mg/L at a signal-to-noise ratio of three. The method developed method was applied to the simultaneous analysis of acidulants and preservatives in a wide variety of food samples.

  3. Rapid differentiation of commercial juices and blends by using sugar profiles obtained by capillary zone electrophoresis with indirect UV detection.

    Science.gov (United States)

    Navarro-Pascual-Ahuir, María; Lerma-García, María Jesús; Simó-Alfonso, Ernesto F; Herrero-Martínez, José Manuel

    2015-03-18

    A method for the determination of sugars in several fruit juices and nectars by capillary zone electrophoresis with indirect UV-vis detection has been developed. Under optimal conditions, commercial fruit juices and nectars from several fruits were analyzed, and the sugar and cyclamate contents were quantified in less than 6 min. A study for the detection of blends of high-value juices (orange and pineapple) with cheaper alternatives was also developed. For this purpose, different chemometric techniques, based on sugar content ratios, were applied. Linear discriminant analysis showed that fruit juices can be distinguished according to the fruit type, juice blends also being differentiated. Multiple linear regression models were also constructed to predict the adulteration of orange and pineapple juices with grape juice. This simple and reliable methodology provides a rapid analysis of fruit juices of economic importance, which is relevant for quality control purposes in food industries and regulatory agencies.

  4. Selective enzymatic cleavage and labeling for sensitive capillary electrophoresis laser-induced fluorescence analysis of oxidized DNA bases.

    Science.gov (United States)

    Li, Cuiping; Wang, Hailin

    2015-08-01

    Oxidatively generated DNA damage is considered to be a significant contributing factor to cancer, aging, and age-related human diseases. It is important to detect oxidatively generated DNA damage to understand and clinically diagnosis diseases caused by oxidative damage. In this study, using selective enzymatic cleavage and quantum dot (QD) labeling, we developed a novel capillary electrophoresis-laser induced fluorescence method for the sensitive detection of oxidized DNA bases. First, oxidized DNA bases are recognized and removed by one DNA base excision repair glycosylase, leaving apurinic and apyrimidinic sites (AP sites) at the oxidized positions. The AP sites are further excised by the AP nicking activity of the chosen glycosylase, generating a nucleotide gap with 5'- and 3'- phosphate groups. After dephosphorylation with one alkaline phosphatase, a biotinylated ddNTP is introduced into the nucleotide space within the DNA strand by DNA polymerase I. The biotin-tagged DNA is further labeled with a QD-streptavidin conjugate via non-covalent interactions. The DNA-bound QD is well-separated from excess DNA-unbound QD by highly efficient capillary electrophoresis and is sensitively detected by online coupled laser-induced fluorescence analysis. Using this method, we can assess the trace levels of oxidized DNA bases induced by the Fenton reaction and UV irradiation. Interestingly, the use of the formamidopyrimidine glycosylase (FPG) protein and endonuclease VIII enables the detection of oxidized purine and pyrimidine bases, respectively. Using the synthesized standard DNA, the approach has low limits of detection of 1.1×10(-19)mol in mass and 2.9pM in concentration. PMID:26105778

  5. Comparative investigations of T cell receptor gamma gene rearrangements in frozen and formalin-fixed paraffin wax-embedded tissues by capillary electrophoresis

    DEFF Research Database (Denmark)

    Christensen, M; Funder, A D; Bendix, K;

    2006-01-01

    AIM: To compare clonal T cell receptor gamma (TCRgamma) gene rearrangements in frozen and formalin-fixed paraffin wax-embedded (FFPE) tissue, using capillary electrophoresis for use in diagnostics, as T cell lymphomas may be difficult to diagnose by conventional methods. METHODS: The DNA for PCR...

  6. A method for studies on interactions between a gold-based drug and plasma proteins based on capillary electrophoresis with inductively coupled plasma mass spectrometry detection

    DEFF Research Database (Denmark)

    Nguyen, Tam T T N; Østergaard, Jesper; Gammelgaard, Bente

    2015-01-01

    An analytical method based on capillary electrophoresis (CE) and inductively coupled plasma mass spectrometry (ICP-MS) detection was developed for studies on the interaction of gold-containing drugs and plasma proteins using auranofin as example. A detection limit of 18 ng/mL of auranofin...

  7. Eletroforese capilar acoplada à espectrometria com plasma: uma ferramenta eficiente para a especiação Capillary electrophoresis coupled to plasma spectrometry: an efficient tool for speciation

    Directory of Open Access Journals (Sweden)

    Ana Paula G. Gervasio

    2003-01-01

    Full Text Available The most important features of the CE-ICP hyphenation, as well as its advantages and drawbacks as a tool for speciation are discussed. The fundamental principles of capillary electrophoresis and inductively coupled plasma mass spectrometry are also presented. Some applications involving different designs proposed in the literature to couple CE and ICP system for elemental speciation are reviewed.

  8. Carboxymethyl-β-cyclodextrin for Chiral Separation of Amino Acids Derivatized with Fluorescene-5-isothiocyanate by Capillary Electrophoresis and Laser-induced Fluorescence Detection

    Institute of Scientific and Technical Information of China (English)

    Yu Yun CHEN; Wei WANG; Wei Ping YANG; Zhu Jun ZHANG

    2004-01-01

    A method using carboxymethyl-β-cyclodextrin(CM-β-CD) as selector for chiral separation of amino acids by capillary electrophoresis and laser-induced fluorescence detection was studied. Resolution was better than that obtained byβ-CD or HP-β-CD.

  9. Analysis of Amino Acids in a Single Human Red Blood Cell by Capillary Zone Electrophoresis with Intracellular NDA derivatization and Electrochemical Detection

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    A novel method for detcrmination of amino acids in individual human red blood cells has been dcveloped. In this method, the derivatization reagents (NDA and CN-) are introduced into living cells by clcctroporation. After completion of derivatization, the amino acids in a single cell is determined by capillary zone electrophoresis with end-column ampcrometric detection.

  10. Analysis of Amino Acids in a Single Human Red Blood Cell by Capillary Zone Electrophoresis with Intracellular NDA—derivatization and Electrochemical Detection

    Institute of Scientific and Technical Information of China (English)

    QianDONG; XiaoLeiWANG; 等

    2002-01-01

    A novel method for determination of amino acids in individual red blood cells has been developed. In this method, the derivatization reagents (NDA and CN-) are introduced into living cells by electroporation. After completion of derivatization,the amino acids in a single cell is determined by capillary zone electrophoresis with end-column amperometric detection.

  11. Modelling of conditions for the enantiomeric separation of beta(2)-adrenergic sympathicomimetics by capillary electrophoresis using cyclodextrins as chiral selectors in a polyethylene glycol gel

    NARCIS (Netherlands)

    de Boer, T; Bijma, R; Ensing, K

    1999-01-01

    A two-factor central composite design was used to determine a mathematical model for prediction of the optimal conditions for the separation of the enantiomers of some widely used beta(2)-sympathicomimetic drugs (beta(2)-agonists) by capillary electrophoresis using cyclodextrins (CD) as a chiral sel

  12. Eletroforese capilar acoplada à espectrometria com plasma: uma ferramenta eficiente para a especiação Capillary electrophoresis coupled to plasma spectrometry: an efficient tool for speciation

    OpenAIRE

    Ana Paula G. Gervasio; André F. Lavorante; Maria Carolina B. Moraes; Maria Fernanda Giné; Carlos E. S. Miranda; Emanuel Carrilho

    2003-01-01

    The most important features of the CE-ICP hyphenation, as well as its advantages and drawbacks as a tool for speciation are discussed. The fundamental principles of capillary electrophoresis and inductively coupled plasma mass spectrometry are also presented. Some applications involving different designs proposed in the literature to couple CE and ICP system for elemental speciation are reviewed.

  13. 6-O-(Hydroxypropyltrimethylammonia)-β-cyclodextrin with Low Degree of Substitution: Convenient Preparation and its Application as a Chiral Selector in Capillary Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    A cationic cyclodextrin derivative 6-O-(hydroxypropyltrimethylammonia)-β-cyclodextrin (GTA-β-CD) with low degree of substitution was prepared through a convenient method in solid phase. The product could be used as a valuable chiral selector in the capillary electrophoresis (CE) separation of some acidic drug enantiomers such as naproxen, ofloxacin,ibuprofen and warfarin.

  14. Voltage-programming-based capillary gel electrophoresis for the fast detection of angiotensin-converting enzyme insertion/deletion polymorphism with high sensitivity.

    Science.gov (United States)

    Woo, Nain; Kim, Su-Kang; Kang, Seong Ho

    2016-08-01

    A voltage-programming-based capillary gel electrophoresis method with a laser-induced fluorescence detector was developed for the fast and highly sensitive detection of DNA molecules related to angiotensin-converting enzyme insertion/deletion polymorphism, which has been reported to influence predisposition to various diseases such as cardiovascular disease, high blood pressure, myocardial infarction, and Alzheimer's disease. Various voltage programs were investigated for fast detection of specific DNA molecules of angiotensin-converting enzyme insertion/deletion polymorphism as a function of migration time and separation efficiency to establish the effect of voltage strength to resolution. Finally, the amplified products of the angiotensin-converting enzyme insertion/deletion polymorphism (190 and 490 bp DNA) were analyzed in 3.2 min without losing resolution under optimum voltage programming conditions, which were at least 75 times faster than conventional slab gel electrophoresis. In addition, the capillary gel electrophoresis method also successfully applied to the analysis of real human blood samples, although no polymorphism genes were detected by slab gel electrophoresis. Consequently, the developed voltage-programming capillary gel electrophoresis method with laser-induced fluorescence detection is an effective, rapid analysis technique for highly sensitive detection of disease-related specific DNA molecules.

  15. Use of metal complexation in non-aqueous capillary electrophoresis systems for the separation and improved detection of tetracyclines.

    Science.gov (United States)

    Tjørnelund, J; Hansen, S H

    1997-08-29

    Metal complexation in non-aqueous capillary electrophoresis systems was evaluated for the separation and improved detection of tetracycline antibiotics using laser-induced fluorescence detection. It was found that three factors were important for the choice of complexing agent: (i) it should be soluble in the organic solvent used for the separation, (ii) it should have a sufficient fast complexing rate so as not to invalidate the electrophoretic separation and, (iii) it should give a large increase in the fluorescence intensity. Mg2+ ions were found to be the most suitable ions for the separation of the tetracyclines as the acetate salt of magnesium is very soluble in organic solvents and only a relatively low current was generated during electrophoresis making it possible to use high concentrations of the complexing metal ion. Metal complexation strongly intensified the fluorescence of tetracyclines and all organic solvents investigated further intensified the fluorescence, e.g. dimethylformamide improved the fluorescence of the oxytetracycline metal complex by a factor of 34 compared to water. However, magnesium acetate was not sufficiently soluble in dimethylformamide and therefore N-methylformamide, improving the fluorescence intensity by only a factor of 9, was used. It was demonstrated that the method can be used for the detection of tetracyclines at the ppb level in milk and plasma. PMID:9335125

  16. Analysis of a ribonuclease H digestion of N3'-->P5' phosphoramidate-RNA duplexes by capillary gel electrophoresis.

    Science.gov (United States)

    DeDionisio, L; Gryaznov, S M

    1995-07-01

    Phosphodiester oligonucleotides (ODNs) and their analogs are presently being investigated as potential antisense therapeutics in the treatment of viral infections and various forms of cancer. here, we would like to report results from an investigation of activity for a ribonuclease H (RNase H) mediated RNA digestion assay in the duplexes formed by an ODN or the ODN analog, N3'-->P5' phosphoramidate (3'-phosphoramidate), and complimentary RNA strands. Capillary gel electrophoresis (CGE) proved to be an effective method for determining RNA hydrolysis in the presence of RNase H. RNA and an ODN or RNA and a 3'-phosphoramidate were hybridized in a Tris-HCl, MgCl2 buffer at room temperature (RT) and incubated with RNase H. Digestions were carried out at RT or at 37 degrees C. Control samples were unhybridized RNA with RNase H, RNA without RNase H, and duplexes (RNA-ODN or 3'-phosphoramidate) without RNase H. All controls were incubated in Tris-HCl, MgCl2 buffer, and sample aliquots were analyzed at various time intervals. A homodecamer, (dT)10, was used as an internal standard to determine the relative migration time of the RNA strand. The final digestion products for the duplexes and the various controls were monitored by CGE. In addition, polyacrylamide gel electrophoresis (PAGE) was used in conjunction with Stains-All (staining) and a densitometric analysis to verify CGE results. PMID:7581876

  17. Study of the interactions between fluoroquinolones and human serum albumin by affinity capillary electrophoresis and fluorescence method

    Energy Technology Data Exchange (ETDEWEB)

    Zhang Liwei; Wang Kun [Beijing National Laboratory for Molecular Sciences (BNLMS), College of Chemistry, Peking University, Beijing 100871 (China); Zhang Xinxiang [Beijing National Laboratory for Molecular Sciences (BNLMS), College of Chemistry, Peking University, Beijing 100871 (China)], E-mail: zxx@pku.edu.cn

    2007-11-05

    The interactions between fluoroquinolones and human serum albumin (HSA) were investigated by affinity capillary electrophoresis (ACE) and fluorescence quenching technique. Based on the efficient separation of several fluoroquinolones using a simple phosphate buffer, the binding constants of fluoroquinolones with HSA were determined simultaneously during one set of electrophoresis by ACE method. The thermodynamic parameters were obtained from data at different temperatures, and the negative {delta}H and {delta}S values showed that both hydrogen bonds and van der Waals interaction played major roles in the binding of fluoroquinolones to HSA. The interactions were also studied by fluorescence quenching technique. The results of fluorescence titration revealed that fluoroquinolones had the strong ability to quenching the intrinsic fluorescence of HSA through the static quenching procedure. The binding site number n, apparent binding constant K{sub b} and the Stern-Volmer quenching constant K{sub sv} were determined. The thermodynamic parameters were also studied by fluorescence method, and the results were consonant with that of ACE.

  18. Development of a capillary electrophoresis method for analyzing adenosine deaminase and purine nucleoside phosphorylase and its application in inhibitor screening.

    Science.gov (United States)

    Qi, Yanfei; Li, Youxin; Bao, James J

    2016-08-01

    A novel capillary electrophoresis (CE) method was developed for simultaneous analysis of adenosine deaminase (ADA) and purine nucleoside phosphorylase (PNP) in red blood cells (RBCs). The developed method considered and took advantage of the natural conversion from the ADA product, inosine to hypoxanthine. The transformation ratio was introduced for ADA and PNP analysis to obtain more reliable results. After optimizing the enzymatic incubation and electrophoresis separation conditions, the determined activities of ADA and PNP in 12 human RBCs were 0.237-0.833 U/ml and 9.013-10.453 U/ml packed cells, respectively. The analysis of ADA in mice RBCs indicated that there was an apparent activity difference between healthy and hepatoma mice. In addition, the proposed method was also successfully applied in the inhibitor screening from nine traditional Chinese medicines, and data showed that ADA activities were strongly inhibited by Rhizoma Chuanxiong and Angelica sinensis. The inhibition effect of Angelica sinensis on ADA is first reported here and could also inhibit PNP activity. PMID:27173606

  19. Developments in coupled solid-phase extraction-capillary electrophoresis 2011-2013.

    Science.gov (United States)

    Ramautar, Rawi; Somsen, Govert W; de Jong, Gerhardus J

    2014-01-01

    This article presents an overview of the design and application of coupled SPE-CE systems that have been reported in the literature between January 2011 and June 2013. The present paper is an update of three previous review papers covering the years 2000-2011 (Electrophoresis 2008, 29, 108-128; Electrophoresis 2010, 31, 44-54; Electrophoresis 2012, 33, 243-250). The use of in-line and on-line SPE-CE approaches is described in this review. Emerging technological developments, such as the use of in-line frit-free SPE and chip-based SPE for extraction of sample components prior to CE analysis, are outlined. Selected examples illustrate the applicability of SPE-CE in biomedical, pharmaceutical, and environmental analysis. A complete overview of recent SPE-CE studies is given in table format, providing information on sample type, SPE sorbent, coupling mode, detection mode, and LOD. Finally, some general conclusions and future perspectives are provided. PMID:24114847

  20. Separation and determination of aloperine, sophoridine, matrine and oxymatrine by combination of flow injection with microfluidic capillary electrophoresis.

    Science.gov (United States)

    Cheng, Yuqiao; Chen, Hongli; Li, Yuqin; Chen, Xingguo; Hu, Zhide

    2004-05-28

    A novel, rapid and accurate method for the separation and determination of aloperine (ALP), sophoridine (SRI), matrine (MT) and oxymatrine (OMT) has been developed by combination of flow injection (FI) with microfluidic capillary electrophoresis (CE) for the first time. In the present paper, a continuous sample introduction interface was described. The interface with an H-channel structure was produced using a non-lithographic approach. The H-channel structure was fixed on a planar plastic base utilizing a horizontal 6.5cm-long separation capillary with two vertical sidearm tubes on each end that served as inlet and outlet flow-through electrode reservoirs. The inlet reservoir also functioned as interface for coupling to the FI system. The buffer solution used was a 50mmoll(-1) borate solution with the pH adjusted to 8.80 with 2moll(-1) HCl. The performance of the system was demonstrated in the separation and determination of ALP, SRI, MT and OMT with UV detection at 215nm, achieving baseline separation within 2min. A series of samples was injected repeatedly without current interruption and subsequent rinsing, and the contents of these four bio-alkaloids in two marketed drugs were determined with satisfactory recovery by this proposed method.

  1. [Rapid determination of lactose, sucrose, glucose and fructose in foods by capillary zone electrophoresis with indirect ultraviolet detection].

    Science.gov (United States)

    Zhang, Huanhuan; Li, Jiang; Zhao, Shan; Ding, Xiaojing; Wang, Zhi

    2015-08-01

    A new and rapid method for the simultaneous determination of lactose, sucrose, glucose and fructose by capillary zone electrophoresis ( CZE) with indirect ultraviolet detection was developed. The separation was completed with an uncoated fused-silica capillary with 30.2 cm of total length (effective length of 20 cm) x 50 µm. The separation buffer consisted of 4 mmol/L potassium sorbate, 10 mmol/L sodium phosphate, 30 mmol/L NaOH (pH 12. 56) and 0. 5 mmol/L hexadecytrimethylammonium bromide (CTAB). The separation was performed at a voltage of -8 kV with the ultraviolet detection at 254 nm. The analysis of the four carbohydrates was completed within 10 min. The limits of detection (S/N= 3) for lactose, sucrose, glucose and fructose were 50, 75, 25 and 25 mg/L, and the limits of quantification (S/N = 10) were 150, 225, 75 and 75 mg/L, respectively. The average recoveries for the four carbohydrates were in the range of 87.0%-107.0% with the relative standard deviations of 1.2%-4.7%. No organic solvent was consumed throughout the whole process of the analysis. The method was used for the analysis of nine food samples and a quality control sample. The results demonstrated that the method is simple, rapid, accurate, and suitable for the routine analysis of the four carbohydrates in food samples.

  2. [Rapid determination of lactose, sucrose, glucose and fructose in foods by capillary zone electrophoresis with indirect ultraviolet detection].

    Science.gov (United States)

    Zhang, Huanhuan; Li, Jiang; Zhao, Shan; Ding, Xiaojing; Wang, Zhi

    2015-08-01

    A new and rapid method for the simultaneous determination of lactose, sucrose, glucose and fructose by capillary zone electrophoresis ( CZE) with indirect ultraviolet detection was developed. The separation was completed with an uncoated fused-silica capillary with 30.2 cm of total length (effective length of 20 cm) x 50 µm. The separation buffer consisted of 4 mmol/L potassium sorbate, 10 mmol/L sodium phosphate, 30 mmol/L NaOH (pH 12. 56) and 0. 5 mmol/L hexadecytrimethylammonium bromide (CTAB). The separation was performed at a voltage of -8 kV with the ultraviolet detection at 254 nm. The analysis of the four carbohydrates was completed within 10 min. The limits of detection (S/N= 3) for lactose, sucrose, glucose and fructose were 50, 75, 25 and 25 mg/L, and the limits of quantification (S/N = 10) were 150, 225, 75 and 75 mg/L, respectively. The average recoveries for the four carbohydrates were in the range of 87.0%-107.0% with the relative standard deviations of 1.2%-4.7%. No organic solvent was consumed throughout the whole process of the analysis. The method was used for the analysis of nine food samples and a quality control sample. The results demonstrated that the method is simple, rapid, accurate, and suitable for the routine analysis of the four carbohydrates in food samples. PMID:26749857

  3. Sensitive simultaneous determination of three sulfanilamide artificial sweeters by capillary electrophoresis with on-line preconcentration and contactless conductivity detection.

    Science.gov (United States)

    Yang, Lirong; Zhou, ShengJi; Xiao, Yuezhou; Tang, Yufeng; Xie, Tianyao

    2015-12-01

    A sensitive method followed by capillary electrophoresis with on-line perconcentration and capacitively coupled contactless conductivity detection (CE-C(4)D) was evaluated as a novel approach for the determination of three sulfanilamide artificial sweeteners (acesulfame-K, sodium saccharin and sodium cyclamate) in beverages. The on-line preconcentration technique, namely field-amplified sample injection, coupled with CE-C(4)D were successfully developed and optimized. The separation was achieved within 10 min under the following conditions: an uncoated fused-silica capillary (45 cm × 50 μm i.d., Leff=40 cm), 20 mmol L(-1) HAc as running buffer, separation voltage of -12 kV, electrokinetic injection of -11 kV × 8 s. The detection limits of acesulfame-K, sodium saccharin and sodium cyclamate were 4.4, 6.7 and 8.8 μg L(-1), respectively. The relative standard deviation varied in the range of 3.0-5.0%. Results of this study show a great potential method for the fast screening of these artificial sweeteners contents in commercial beverages. PMID:26041216

  4. Capillary zone electrophoresis for analysis of phytochelatins and other thiol peptides in complex biological samples derivatized with monobromobimane.

    Science.gov (United States)

    Perez-Rama, Mónica; Torres Vaamonde, Enrique; Abalde Alonso, Julio

    2005-02-01

    A new method to improve the analysis of phytochelatins and their precursors (cysteine, gamma-Glu-Cys, and glutathione) derivatized with monobromobimane (mBrB) in complex biological samples by capillary zone electrophoresis is described. The effects of the background electrolyte pH, concentration, and different organic additives (acetonitrile, methanol, and trifluoroethanol) on the separation were studied to achieve optimum resolution and number of theoretical plates of the analyzed compounds in the electropherograms. Optimum separation of the thiol peptides was obtained with 150 mM phosphate buffer at pH 1.60. Separation efficiency was improved when 2.5% v/v methanol was added to the background electrolyte. The electrophoretic conditions were 13 kV and capillary dimensions with 30 cm length from the inlet to the detector (38 cm total length) and 50 microm inner diameter. The injection was by pressure at 50 mbar for 17 s. Under these conditions, the separation between desglycyl-peptides and phytochelatins was also achieved. We also describe the optimum conditions for the derivatization of biological samples with mBrB to increase electrophoretic sensitivity and number of theoretical plates. The improved method was shown to be simple, reproducible, selective, and accurate in measuring thiol peptides in complex biological samples, the detection limit being 2.5 microM glutathione at a wavelength of 390 nm.

  5. Determination of heroin metabolites in human urine using capillary zone electrophoresis with beta-cyclodextrin and UV detection.

    Science.gov (United States)

    Alnajjar, Ahmed; McCord, Bruce

    2003-10-15

    A method has been developed for the detection of a mixture of morphine, codeine, 6-acetyl morphine (6-AM) and normorphine using capillary zone electrophoresis (CZE). The method utilized urinary 6-AM as a diagnostic indicator of heroin abuse because it is not a product of either morphine or codeine metabolism. The electrophoretic separation was achieved using an uncoated (50 microm I.D.) fused-silica capillary, 77 cm long, containing the detector window 10.0 cm from the outlet end. The running buffer (pH 6.0) contained 50 mM sodium phosphate and 0.015 M beta-cyclodextrins (beta-CD). The samples were first extracted using a mixed-mode solid-phase extraction procedure and then analyzed by CZE. The UV absorbance detection was monitored at 214 nm. It has been found that beta-CDs can improve separation efficiency due to their hydrophobic cavity. The effect of the concentration of beta-CD and pH was also evaluated. The application of electrokinetic injection with field amplified sample stacking results in low detection limits (40 ng/ml for each analyte) and the method has good reproducibility, precision, accuracy, and high recovery. PMID:14550865

  6. Separation and quantification of viral double-stranded RNA fragments by capillary electrophoresis in hydroxyethylcellulose polymer solutions.

    Science.gov (United States)

    Shambaugh, C L; Bodmer, J L; Hsu, D; Ranucci, C S

    2004-10-01

    Capillary electrophoresis (CE) is an analytical technique widely utilized to resolve complex mixtures of nucleic acids. CE uses a variety of polymers in solution that act as a molecular sieve to separate nucleic acid fragments according to size. It has been shown previously that purified dsDNA can be resolved efficiently by solutions of hydroxyethylcellulose (HEC) polymer, providing a rapid and high resolution method of separation. We have applied this separation technique to viral double-stranded (ds) RNA segments derived from rotavirus process samples. HEC polymers of various molecular masses and concentrations were identified and compared for their ability to separate dsRNA based on the extent of expected polymer network formation. The HEC polymer exhibiting the most desirable separation characteristics was then used for subsequent optimization of various method parameters, such as, injection time, electric field strength, dye concentration and capillary equilibration. The optimized method was then applied to the quantification of genome concentration based on a representative segment of the rotavirus genome. This study demonstrated that purified viral dsRNA material of known concentration could be used to generate an external standard curve relating concentration to peak area. This standard curve was used to determine the concentration of unknown samples by interpolation. This novel RNA quantification assay is likely to be applicable to other types of virus, including those containing dsDNA.

  7. Validated Method for the Determination of Piroxicam by Capillary Zone Electrophoresis and Its Application to Tablets

    Directory of Open Access Journals (Sweden)

    Arın Gül Dal

    2014-01-01

    piroxicam in tablets. The separation of piroxicam was conducted in a fused-silica capillary by using 10 mM borate buffer (pH 9.0 containing 10% (v/v methanol as background electrolyte. The optimum conditions determined were 25 kV for separation voltage and 1 s for injection time. Analysis was carried out with UV detection at 204 nm. Naproxen sodium was used as an internal standard. The method was linear over the range of 0.23–28.79 µg/mL. The accuracy and precision were found to be satisfied within the acceptable limits (<2%. The LOD and LOQ were found to be 0.07 and 0.19 µg/mL, respectively. The method described here was applied to tablet dosage forms and the content of a tablet was found in the limits of USP-24 suggestions. To compare the results of capillary electrophoretic method, UV spectrophotometric method was developed and the difference between two methods was found to be insignificant. The capillary zone electrophoretic method developed in this study is rapid, simple, and suitable for routine analysis of piroxicam in pharmaceutical tablets.

  8. Determination of polyphenols in Spanish wines by capillary zone electrophoresis. Application to wine characterization by using chemometrics.

    Science.gov (United States)

    Franquet-Griell, Helena; Checa, Antonio; Núñez, Oscar; Saurina, Javier; Hernández-Cassou, Santiago; Puignou, Lluis

    2012-08-29

    A capillary zone electrophoresis (CZE) method for the simultaneous determination of 20 polyphenols in wine was developed. The separation was performed using fused-silica capillaries of 75 μm i.d. and a 30 mM sodium tretraborate buffer solution at pH 9.2 with 5% isopropanol as a background electrolyte. A capillary voltage of +25 kV with pressure-assisted (3.5 kPa) separation from minute 18 was applied, thus achieving a total analysis time of 0.990), and run-to-run and day-to-day precisions (RSD values lower than 6.5 and 15.7%, respectively) were established. Three different calibration procedures were evaluated for polyphenol quantitation in wines: external calibration using standards prepared in Milli-Q water, standard addition, and pseudomatrix-matched calibration using wine as a matrix. For a 95% confidence level, no statistical differences were observed, in general, between the three calibration methods (p values between 0.11 and 0.84), whereas for some specific polyphenols, such as cinnamic acid, syringic acid, and gallic acid, results were not comparable when external calibration was used. The CZE method using pseudomatrix-matched calibration was then proposed and applied to the analysis of polyphenols in 49 Spanish wines, showing satisfactory results and a wide compositional variation between wines. Electrophoretic profiles and other compositional data (e.g., peak areas of selected peaks) were considered as fingerprints of wines to be used for characterization and classification purposes. The corresponding data were analyzed by principal component analysis (PCA) to extract information on the most significant features contributing to wine discrimination according to their origins. Results showed that a reasonable distribution of wines depending on the elaboration areas was found, tyrosol and gallic, protocatechuic, p-coumaric, and caffeic acids being some representative discriminant compounds.

  9. Electrokinetic supercharging in nonaqueous capillary electrophoresis for online preconcentration and determination of tamoxifen and its metabolites in human plasma.

    Science.gov (United States)

    Thang, Lee Yien; Breadmore, Michael C; See, Hong Heng

    2016-08-26

    An online preconcentration method, namely electrokinetic supercharging (EKS), was evaluated for the determination of tamoxifen and its metabolites in human plasma in nonaqueous capillary electrophoresis with ultraviolet detection (NACE-UV). This method was comprehensively optimized in terms of the leading electrolyte (LE) and terminating electrolyte (TE) injection lengths, as well as electrokinetic sample injection time. The optimized EKS conditions employed were as follows: hydrodynamic injection (HI) of 10mM potassium chloride as LE at 150mbar for 36s (4% of total capillary volume). The sample was injected at 10kV for 300s, followed by HI of 10mM pimozide as TE at 150mbar for 36s (4% of total capillary volume). Separation was performed in 7.5mM deoxycholic acid sodium salt, 15mM acetic acid and 1mM 18-crown-6 in 100% methanol at +25kV with UV detection at 205nm. Under optimized conditions, the sensitivity was enhanced between 160- and 600-fold when compared with our previously developed method based on HI at 150mbar for 12s. The detection limit of the method for tamoxifen and its metabolites were 0.05-0.25ng/mL, with RSDs between 2.1% and 3.5%. Recoveries in spiked human plasma were 95.6%-99.7%. A comparison was also made between the proposed EKS approach and the standard field-amplified sample injection (FASI) technique. EKS proved to be 3-5 times more sensitive than the FASI. The new EKS method was applied to the analysis of tamoxifen and its metabolites in plasma samples from breast cancer patients after liquid-liquid extraction. PMID:27485148

  10. The analysis of cations and anions in illicit heroin using capillary electrophoresis with indirect UV detection.

    Science.gov (United States)

    Lurie, I S

    1996-01-01

    Methodology is presented for the analysis of cations and anions in illicit heroin using CE with indirect UV detection. The cations investigated include ammonium, calcium, potassium, magnesium, and sodium; the anions included acetate, chloride, citrate, phosphate, sulfate, and tartrate. For cations, the Ion Phor run buffer (Dionex Corp., Sunnyvale, CA, U.S.A.) consisting of 4 mM copper sulfate, 4 mM formic acid, and 3 mM 18-crown-6 (pH 3.0) was used. For anions, proprietary reagents were used, including the Anitron run buffer (PE Applied Biosystems, Foster City, CA, U.S.A.) and Micro-Coat capillary charge-reversal agent (PE Applied Biosystems), which was utilized to flush the capillary prior to each analysis. Lithium nitrate was used as an internal standard; excellent long- and short-term precision in relative retention times were obtained for both cations and anions. The short-term precision in peak areas was satisfactory. For the various ions examined, a linearity range of a little less than two orders of magnitude was observed. The methodology is capable of analyzing ions down to the 10(-3)% level relative to heroin.

  11. Optoelectronic Capillary Sensors in Microfluidic and Point-of-Care Instrumentation

    Directory of Open Access Journals (Sweden)

    Michał Borecki

    2010-04-01

    Full Text Available This paper presents a review, based on the published literature and on the authors’ own research, of the current state of the art of fiber-optic capillary sensors and related instrumentation as well as their applications, with special emphasis on point-of-care chemical and biochemical sensors, systematizing the various types of sensors from the point of view of the principles of their construction and operation. Unlike classical fiber-optic sensors which rely on changes in light propagation inside the fiber as affected by outside conditions, optical capillary sensors rely on changes of light transmission in capillaries filled with the analyzed liquid, which opens the possibility of interesting new applications, while raising specific issues relating to the construction, materials and instrumentation of those sensors.

  12. Effects of improved microchannel structures on the separation characteristics of microchip capillary electrophoresis

    CERN Document Server

    Utsumi, Y; Ozaki, M; Terabe, S

    2003-01-01

    We fabricated the electrophoresis microchips using the UV polymerization technique. We employed plastic substrates that were suitable for rapid prototyping instead of glass and quartz. A thick UV negative photo resist was used to form molds and poly-dimethylsilozane (PDMS) was polymerized by a thermal curing process on the mold to obtain replica microchips. Electroosmotic flow (EOF) was measured to evaluate the surface. Rhodamine B and sulforhodamine B are successfully separated using the microchip. Characteristic differences between UV-fabricated and SR-fabricated microchips were evaluated by EOF measurement. It was observed that accurately defined microchannels fabricated by synchrotron radiation (SR) lithography show constant peak heights and FWHMs. Thus the advantage of the application of SR lithography to the mold fabrication is also demonstrated. (author)

  13. Electrostatic interaction mechanism on the separation of phenols by non-aqueous capillary electrophoresis

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    The electrostatic interaction between additive and analyte is of great importance to non-aqueous cap- illary electrophoresis(NACE)separation.Three tetraalkylammonium bromides and acetonitrile were applied as additives and running solvent respectively.The effect of alkyl chain length and concentra- tion of additive on electrostatic interactions was investigated by the separation of phenols.The sepa- ration ability was found to increase with decreasing alkyl chain length of the additive,and the resolu- tion values were increased with increasing additive concentration.The separation was seriously dete- riorated after a little amount of water was added in the running solution.Furthermore,the electrostatic interaction is strong under the conditions of low electron cloud density,weak steric hindrance and multi-interaction sites.Thus,the separation result can be predicted by theoretical analysis,which is helpful for the separation of other substances in NACE based on electrostatic interaction.

  14. Evaluation of the performance of single-walled carbon nanohorns in capillary electrophoresis.

    Science.gov (United States)

    Jiménez-Soto, Juan Manuel; Moliner-Martínez, Yolanda; Cárdenas, Soledad; Valcárcel, Miguel

    2010-05-01

    This paper describes for the first time the use of single-walled carbon nanohorns (SWNHs) as pseudostationary and stationary phases for EKC and CEC, respectively, taking advantage of their characteristic features, such as conical-end termination, formation of spherical assemblies dahlia-flower like superstructure and easy functionalization. The use of SWNHs as pseudostationary phase for EKC required the study of their dispersion in different surfactants as well as their compatibility with the electrophoretic system. The carboxylation and subsequent immobilization of carboxylated SWNHs in fused-silica capillary to obtain useful, reproducible and stable stationary phases for CEC has also been investigated, with promising results. The electrophoretic separations obtained for water-soluble vitamins in both modalities (EKC and CEC) have been systematically compared with those obtained with single-walled carbon nanotubes. PMID:20419702

  15. Separations of flavan-3-ols and dimeric proanthocyanidins by capillary electrophoresis.

    Science.gov (United States)

    Kreimeyer, J; Petereit, F; Nahrstedt, A

    1998-02-01

    Micellar electrokinetic capillary chromatography (MECC), using a phosphate buffer pH 7.0 and sodium dodecyl sulfate as the surfactant, was found suitable for the separation of structurally similar flavan-3-ols and related dimeric proanthocyanidins. A total of 20 substances has been tested, 17 of which were successfully separated simultaneously in less than 7 minutes. Retention of the investigated compounds by the SDS micelles is affected by (i) the stereochemistry of the heterocycle of the flavan-3-ol skeleton, (ii) the molecular size, the number and position of free phenolic hydroxy groups, and (iii) the type of acyl residues attached at the C-3 hydroxy function. The method was successfully applied to the phytochemical analysis of tannin enriched extracts of Cistus incanus L. ssp. incanus and C. salvifolius L. (Cistaceae), two traditionally used medicinal plants. PMID:17253220

  16. Determination of Five Organic Acids in Radix Isatidis by Column Partition Chromatography and Capillary Zone Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    CHAIYi-fen; JISong-gang; ZHANGGuo-qing; LIUChang-hai

    2003-01-01

    Aim To determine five organic acids in Radix lsatidis. Method The extraction method and the column partition chromatographic conditiom were studied. Then a capillary zone dectrophorefic method was set up for the determina-tion. Results The linear ranges of quinazolinone acid, n-anthranilic acid, benzoic acid, salicylic acid, and syringic acid were 5.52 - 92.0μg·mL-1 , 5.12 - 102μg·mL-1 , 2.28 - 84.4μg·mL-1, 4.78 - 159 μg·mL-1, and 1.74- 87.0μg·mL-1 respectively. Conclusion The established method is accurate and simple.

  17. Analysis of proteins in biological samples by capillary sieving electrophoresis with postcolumn derivatization/laser-induced fluorescence detection.

    Science.gov (United States)

    Kaneta, Takashi; Ogura, Takehito; Imasaka, Totaro

    2011-04-01

    Previously, we have demonstrated postcolumn derivatization of proteins separated by capillary sieving electrophoresis (CSE), in which naphthalene-2,3-dicarbaldehyde was employed as a fluorogenic labeling reagent. Standard proteins separated by CSE were reacted with naphthalene-2,3-dicarbaldehyde in the presence of 2-mercaptoethanol (2-ME) which plays a role of a reducing agent in the derivatization reaction. To improve the sensitivity, we attempted the use of ethanethiol instead of 2-ME. Ethanethiol showed 1.4- to 4.5-fold lower limits of detection for proteins than 2-ME. Furthermore, we found that 8-aminopyrene-1,3,6-trisulfonate (APTS) is a good marker for relative electrophoretic mobilities of proteins in CSE. Since APTS is a fluorescent and trivalent anion, it generates strong fluorescence and migrates faster than any of the proteins. Therefore, we employed APTS as a marker to obtain the relative electrophoretic mobilities of proteins. The present method was applied to the analyses of proteins in biological samples. Human Ewing's family tumor cell line 'RDES' was used as a sample. The cultured cells were lysed with a buffer containing Tris-HCl, NaCl, sodium dodecyl sulfate, and 2-ME. After denaturation, the lysate was directly introduced into the capillary. Several peaks, which would correspond to proteins with molecular mass ranging from 10 to 93 kDa, were found in the cell lysate. In addition, we measured a milk sample by the CSE with postcolumn derivatization. The electropherogram showed five major peaks which corresponded to α-lactalbumin, β-lactoglobulin, κ-casein, bovine serum albumin, and mixture of α- and β-casein. PMID:21449073

  18. A high-efficiency cross-flow micronebulizer interface for capillary electrophoresis and inductively coupled plasma mass spectrometry.

    Science.gov (United States)

    Li, J; Umemura, T; Odake, T; Tsunoda, K

    2001-12-15

    A pneumatic nebulizer interface for capillary electrophoresis (CE) and inductively coupled plasma mass spectrometry (ICPMS) is reported. The interface is constructed using a high-efficiency cross-flow micronebulizer (HECFMN) and has the following features. (1) Makeup solutions can be fed to the interface by nebulizer self-aspiration and liquid gravity pressurization. (2) The liquid dead volume of the interface is approximately 65 nL, much smaller than those (200-2500 nL) reported for other interfaces. (3) The interface can be stably operated at a liquid flow rate down to 5 microL/min with a high analyte transport efficiency up to 95% to the plasma and (4) does not induce noticeable laminar flow in the CE capillary at typical nebulizer gas flow rates of 0.8-1.2 L/min. Because of these features, baseline resolution of 10 lanthanides with a CE-ICPMS system using the HECFMN interface is achieved, and detection limits and peak asymmetry are 0.05-1 microg/L and 0.93-1.23, respectively, improved significantly over those reported previously for a CE-ICPMS system using a high-efficiency nebulizer interface. Peak precision for the 10 lanthanides is in the range of 6.2-12.3% RSD (N = 5). Peak widths are from 9.1 s for 139La to 17.9 s for 175Lu. The effects of nebulizer gas flow rate, makeup solution flow rate, and spray chamber volume on CE-ICPMS signal intensity and separation are also evaluated for the HECFMN interface by the separation of Cr3+ and Cr2O7(2-).

  19. Determination of urinary 8-hydroxy-2′-deoxyguanosine by capillary electrophoresis with molecularly imprinted monolith in-tube solid phase microextraction

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Urinary 8-hydroxy-2'-deoxyguanosine(8-OHdG) is an excellent marker of oxidative DNA damage.In this study,employing guanosine as dummy template a novel molecularly imprinted(MIP) monolithic capillary column had been synthesized,and that was used as medium of in-tube solid phase microextraction(SPME).Coupled with capillary electrophoresis-electrochemical detection(CE-ECD),the system of extraction and detection of 8-OHdG in urinary sample had been developed.Because of its greater phase ratio combined with c...

  20. Drop-by-drop chemical reaction and sample introduction for capillary electrophoresis.

    Science.gov (United States)

    Chen, Fengming; Rang, Ying; Weng, Ying; Lin, Luyao; Zeng, Hulie; Nakajim, Hizuru; Lin, Jin-Ming; Uchiyama, Katsumi

    2015-06-21

    In this paper, we report a novel sample introduction and chemical reaction strategy by drop-by-drop inkjet injection for an electrophoretically mediated microanalysis (EMMA). This method makes it possible to achieve an on-line introduction of reactant solutions by alternately ejecting small plugs, with an overlapping region of the plugs for mixing the reactants by electrophoresis, supporting chemical reactions, followed by electrophoretic separation of the final compounds. As a proof-of-concept of the method, the EMMA of an inkjetted mixture of 4-fluoro-7-nitrobenzofurazan (NBD-F) and amino acids was carried out as a model chemical reaction. The product NBD-amino acids were quantified by detection with laser induced fluorescence. The optimal conditions for the procedure were: inkjet driving voltage: +40-44 V; pulse width: 20-24 μs; drop-by-drop injection of reactant solutions: alternately 2 drops × 25 times for the amino acid solution and the NBD-F solution; zone overlapping voltage and time: 3 kV and 2 s; incubation time after overlapping: 5 min; separation voltage: 18 kV. Under the optimized conditions, a significant enhancement in sensitivity and a sensitive quantitative analysis were realized. The results obtained were comparable with those using the off-line labeling method. This method is rapid, cost-effective, and readily automated for EMMA. PMID:25728632

  1. Online preconcentration of arsenic compounds by dynamic pH junction-capillary electrophoresis.

    Science.gov (United States)

    Jaafar, Jafariah; Irwan, Zildawarni; Ahamad, Rahmalan; Terabe, Shigeru; Ikegami, Tohru; Tanaka, Nobuo

    2007-02-01

    An online preconcentration technique by dynamic pH junction was studied to improve the detection limit for anionic arsenic compounds by CE. The main target compound is roxarsone, or 3-nitro-4-hydroxyphenylarsonic acid, which is being used as an animal feed additive. The other inorganic and organoarsenic compounds studied are the possible biotransformation products of roxarsone. The arsenic species were separated by a dynamic pH junction in a fused-silica capillary using 15 mM phosphate buffer (pH 10.6) as the BGE and 15 mM acetic acid as the sample matrix. CE with UV detection was monitored at a wavelength of 192 nm. The influence of buffer pH and concentration on dynamic pH junction were investigated. The arsenic species focusing resulted in LOD improvement by a factor of 100-800. The combined use of C18 and anion exchange SPE and dynamic pH junction to CE analysis of chicken litter and soils helps to increase the detection sensitivity. Recoveries of spiked samples ranged between 70 and 72%.

  2. A simple and compact fluorescence detection system for capillary electrophoresis and its application to food analysis.

    Science.gov (United States)

    Zhai, Haiyun; Yuan, Kaisong; Yu, Xiao; Chen, Zuanguang; Liu, Zhenping; Su, Zihao

    2015-10-01

    A novel fluorescence detection system for CE was described and evaluated. Two miniature laser pointers were used as the excitation source. A Y-style optical fiber was used to transmit the excitation light and a four-branch optical fiber was used to collect the fluorescence. The optical fiber and optical filter were imported into a photomultiplier tube without any extra fixing device. A simplified PDMS detection cell was designed with guide channels through which the optical fibers were easily aligned to the detection window of separation capillary. According to different requirements, laser pointers and different filters were selected by simple switching and replacement. The fluorescence from four different directions was collected at the same detecting point. Thus, the sensitivity was enhanced without peak broadening. The fluorescence detection system was simple, compact, low-cost, and highly sensitive, with its functionality demonstrated by the separation and determination of red dyes and fluorescent whitening agents. The detection limit of rhodamine 6G was 7.7 nM (S/N = 3). The system was further applied to determine illegal food dyes. The CE system is potentially eligible for food safety analysis. PMID:26109527

  3. High-Throughput Analysis With 96-Capillary Array Electrophoresis and Integrated Sample Preparation for DNA Sequencing Based on Laser Induced Fluorescence Detection

    Energy Technology Data Exchange (ETDEWEB)

    Gang Xue

    2001-12-31

    The purpose of this research was to improve the fluorescence detection for the multiplexed capillary array electrophoresis, extend its use beyond the genomic analysis, and to develop an integrated micro-sample preparation system for high-throughput DNA sequencing. The authors first demonstrated multiplexed capillary zone electrophoresis (CZE) and micellar electrokinetic chromatography (MEKC) separations in a 96-capillary array system with laser-induced fluorescence detection. Migration times of four kinds of fluoresceins and six polyaromatic hydrocarbons (PAHs) are normalized to one of the capillaries using two internal standards. The relative standard deviations (RSD) after normalization are 0.6-1.4% for the fluoresceins and 0.1-1.5% for the PAHs. Quantitative calibration of the separations based on peak areas is also performed, again with substantial improvement over the raw data. This opens up the possibility of performing massively parallel separations for high-throughput chemical analysis for process monitoring, combinatorial synthesis, and clinical diagnosis. The authors further improved the fluorescence detection by step laser scanning. A computer-controlled galvanometer scanner is adapted for scanning a focused laser beam across a 96-capillary array for laser-induced fluorescence detection. The signal at a single photomultiplier tube is temporally sorted to distinguish among the capillaries. The limit of detection for fluorescein is 3 x 10{sup -11} M (S/N = 3) for 5-mW of total laser power scanned at 4 Hz. The observed cross-talk among capillaries is 0.2%. Advantages include the efficient utilization of light due to the high duty-cycle of step scan, good detection performance due to the reduction of stray light, ruggedness due to the small mass of the galvanometer mirror, low cost due to the simplicity of components, and flexibility due to the independent paths for excitation and emission.

  4. Analysis of water from the Space Shuttle and Mir Space Station by ion chromatography and capillary electrophoresis

    Science.gov (United States)

    Orta, D.; Mudgett, P. D.; Ding, L.; Drybread, M.; Schultz, J. R.; Sauer, R. L.

    1998-01-01

    Drinking water and condensate samples collected from the US Space Shuttle and the Russian Mir Space Station are analyzed routinely at the NASA-Johnson Space Center as part of an ongoing effort to verify water quality and monitor the environment of the spacecraft. Water quality monitoring is particularly important for the Mir water supply because approximately half of the water consumed is recovered from humidity condensate. Drinking water on Shuttle is derived from the fuel cells. Because there is little equipment on board the spacecraft for monitoring the water quality, samples collected by the crew are transported to Earth on Shuttle or Soyuz vehicles, and analyzed exhaustively. As part of the test battery, anions and cations are measured by ion chromatography, and carboxylates and amines by capillary electrophoresis. Analytical data from Shuttle water samples collected before and after several missions, and Mir condensate and potable recovered water samples representing several recent missions are presented and discussed. Results show that Shuttle water is of distilled quality, and Mir recovered water contains various levels of minerals imparted during the recovery processes as designed. Organic ions are rarely detected in potable water samples, but were present in humidity condensate samples.

  5. Optimization and validation of a rapid method to determine citrate and inorganic phosphate in milk by capillary electrophoresis.

    Science.gov (United States)

    Izco, J M; Tormo, M; Harris, A; Tong, P S; Jimenez-Flores, R

    2003-01-01

    Quantification of phosphate and citrate compounds is very important because their distribution between soluble and colloidal phases of milk and their interactions with milk proteins influence the stability and some functional properties of dairy products. The aim of this work was to optimize and validate a capillary electrophoresis method for the rapid determination of these compounds in milk. Various parameters affecting analysis have been optimized, including type, composition, and pH of the electrolyte, and sample extraction. Ethanol, acetonitrile, sulfuric acid, water at 50 degrees C or at room temperature were tested as sample buffers (SB). Water at room temperature yielded the best overall results and was chosen for further validation. The extraction time was checked and could be shortened to less than 1 min. Also, sample preparation was simplified to pipet 12 microl of milk into 1 ml of water containing 20 ppm of tartaric acid as an internal standard. The linearity of the method was excellent (R2 > 0.999) with CV values of response factors yogurt, or Cheddar cheese were analyzed and accuracy was similar to milk in all products tested. The procedure is rapid and offers a very fast and simple sample preparation. Once the sample has arrived at the laboratory, less than 5 min (including handling, preparation, running, integration, and quantification) are necessary to determine the concentration of citric acid and inorganic phosphate. Because of the speed and accuracy of this method, it is promising as an analytical quantitative testing technique.

  6. Enantiomeric separation of tapentadol by capillary electrophoresis--study of chiral selectivity manipulation by various types of cyclodextrins.

    Science.gov (United States)

    Znaleziona, Joanna; Fejős, Ida; Ševčík, Juraj; Douša, Michal; Béni, Szabolcs; Maier, Vítězslav

    2015-02-01

    The chiral recognition of the centrally acting analgesic agent tapentadol and its isomers with various cyclodextrins (CDs) was studied by capillary electrophoresis, focusing on the migration order of four stereoisomers. In the case of non-charged hydroxypropylated CDs (2-hydroxypropyl-β-CD, 2-hydroxypropyl-γ-CD) the beta derivative was able to discriminate the S,R- and R,S-isomers in acidic background electrolyte, whereas the gamma allowed the separation of S,S- and R,R-tapentadol, respectively. Dual CD system containing both hosts was used to separate all of four isomers. Negatively charged sulfated-α-CD at 1.0% (w/v) concentration in 100mM sodium borate buffer (pH 9.5) was capable of separating the isomers with favorable enantiomer migration order and the optimized method was able to determine 0.15% of chiral impurities of tapentadol in the presence of the last migrating clinically important R,R-isomer. PMID:25527976

  7. Capillary zone electrophoresis for fatty acids with chemometrics for the determination of milk adulteration by whey addition.

    Science.gov (United States)

    de Oliveira Mendes, Thiago; Porto, Brenda Lee Simas; Bell, Maria José Valenzuela; Perrone, Ítalo Tuler; de Oliveira, Marcone Augusto Leal

    2016-12-15

    Adulteration of milk with whey is difficult to detect because these two have similar physical and chemical characteristics. The traditional methodologies to monitor this fraud are based on the analysis of caseinomacropeptide. The present study proposes a new approach to detect and quantify this fraud using the fatty acid profiles of milk and whey. Fatty acids C14:0, C16:0, C18:0, C18:1, C18:2 and C18:3 were selected by gas chromatography associated with discriminant analysis to differentiate milk and whey, as they are present in quite different amounts. These six fatty acids were quantified within a short time by capillary zone electrophoresis in a set of adulterated milk samples. The correlation coefficient between the true values of whey addition and the experimental values obtained by this technique was 0.973. The technique is thus useful for the evaluation of milk adulteration with whey, contributing to the quality control of milk in the dairy industry. PMID:27451230

  8. Enantiomeric separation of tapentadol by capillary electrophoresis--study of chiral selectivity manipulation by various types of cyclodextrins.

    Science.gov (United States)

    Znaleziona, Joanna; Fejős, Ida; Ševčík, Juraj; Douša, Michal; Béni, Szabolcs; Maier, Vítězslav

    2015-02-01

    The chiral recognition of the centrally acting analgesic agent tapentadol and its isomers with various cyclodextrins (CDs) was studied by capillary electrophoresis, focusing on the migration order of four stereoisomers. In the case of non-charged hydroxypropylated CDs (2-hydroxypropyl-β-CD, 2-hydroxypropyl-γ-CD) the beta derivative was able to discriminate the S,R- and R,S-isomers in acidic background electrolyte, whereas the gamma allowed the separation of S,S- and R,R-tapentadol, respectively. Dual CD system containing both hosts was used to separate all of four isomers. Negatively charged sulfated-α-CD at 1.0% (w/v) concentration in 100mM sodium borate buffer (pH 9.5) was capable of separating the isomers with favorable enantiomer migration order and the optimized method was able to determine 0.15% of chiral impurities of tapentadol in the presence of the last migrating clinically important R,R-isomer.

  9. Separation and quantitation of phycobiliproteins using phytic acid in capillary electrophoresis with laser-induced fluorescence detection.

    Science.gov (United States)

    Viskari, Pertti J; Colyer, Christa L

    2002-10-01

    The similar electrophoretic mobilities and sizes of several of the phycobiliproteins, which are derived from the photosynthetic apparatus of cyanobacteria and eukaryotic algae, render their separation and quantitation a challenging problem. However, we have developed a suitable capillary electrophoresis (CE) method that employs a phytic acid-boric acid buffer and laser-induced fluorescence (LIF) detection with a single 594 nm He-Ne laser. This method takes advantage of the remarkably high quantum yields of these naturally fluorescent proteins, which can be attributed to their linear tetrapyrrole chromophores covalently bound to cysteinyl residues. As such, limits of detection of 1.18 x 10(-14), 5.26 x 10(-15), and 2.38 x 10(-15) mol/l were obtained for R-phycoerythrin, C-phycocyanin, and allophycocyanin proteins, respectively, with a linear dynamic range of eight orders of magnitude in each case. Unlike previously published CE-LIF methods, this work describes the separation of all three major classes of phycobiliproteins in under 5 min. Very good recoveries, ranging from 93.2 to 105.5%, were obtained for a standard mixture of the phycobiliproteins, based on seven-point calibration curves for both peak height and peak area. It is believed that this development will prove useful for the determination of phycobiliprotein content in naturally occurring cyanobacteria populations, thus providing a useful tool for understanding biological and chemical oceanographic processes.

  10. DNA mutation detection with chip-based temperature gradient capillary electrophoresis using a slantwise radiative heating system.

    Science.gov (United States)

    Zhang, Hui-Dan; Zhou, Jing; Xu, Zhang-Run; Song, Jin; Dai, Jing; Fang, Jin; Fang, Zhao-Lun

    2007-09-01

    A simple and robust chip-based temperature gradient capillary electrophoresis (TGCE) system was developed for DNA mutation/single-nucleotide polymorphism (SNP) analysis using a radiative heating system. Reproducible, stable and uniform temperature gradients were established along a 3 cm length of the electrophoretic separation channel using a single thermostated aluminium heater plate. The heater was slightly slanted relative to the plane of the glass chip at 0.2-1.3 degrees by inserting thin spacers between the plate and chip at one end to produce differences in radiative heating that created the temperature gradient. On-chip TGCE analyses of 4 mutant DNA model samples amplified from plasmid templates, each containing a single base substitution, with a wide range of melting temperatures, showed that mutations were successfully detected under a wide temperature gradient of 10 degrees C and within a short gradient region of about 3 cm (3.3 degrees C cm(-1) gradient). The radiative heating system was able to establish stable spatial temperature gradients along short microfluidic separation channels using simple peripheral equipment and manipulation while ensuring good resolution for detecting a wide range of mutations. Effectiveness of the system was demonstrated by the successful detection of K-ras gene mutations in 6 colon cancer cell lines.

  11. Separation of Key Biogenic Amines by Capillary Electrophoresis and Determination of Possible Indicators of Sport Fatigue in Athlete's Urine.

    Science.gov (United States)

    He, Lili; Ren, Jie; Shi, Zhihao; Xu, Zhongqi

    2016-09-01

    This article aims to build up a simple, rapid and accurate capillary zone electrophoresis (CZE) method for the separation of biogenic amines (BAs). Here, 10 key BAs (phenethylamine, histamine, tryptamine, tyramine, 5-hydroxytryptamine, octopamine, dopamine, norepinephrine, epinephrine and carnosine) owning significant functions were chosen for method development. The baseline separation and identification of 10 standards of the mixture by CZE were eventually achieved in 150.0 mmol/L phosphate buffer (Na2HPO4-NaH2PO4) containing 1.0 mmol/L borax at a pH of 6.1. The addition of borax was found effective for improving the isomeric separation of octopamine and dopamine. The proposed method allowed the limits of detections of BAs to be in the range of 0.2-1.2 µmol/L at UV detection (200 nm); the relative standard deviation of the migration time and the peak area were in the ranges 0.08-0.12 and 2.74-4.63% (n = 5), respectively. Formaldehyde (a possible antiseptic in urine) and five main matrices in urine were studied for the identification of BAs. Finally, profiling of BAs in actual urine from athletes was carried out. Currently, only phenethylamine, norepinephrine and carnosine were designated by spiking the standards. In addition, their variation in athletes' urine has been checked at different states of sport fatigue with the goal of obtaining possible indicators of sport fatigue. PMID:27139740

  12. Fast Determination of Clenbuterol and Salbutamol in Feed and Meat Products Based on Miniaturized Capillary Electrophoresis with Amperometric Detection

    Institute of Scientific and Technical Information of China (English)

    CHU Qing-Cui; GENG Cheng-Huai; ZHOU Hui; YE Jian-Nong

    2007-01-01

    The fast separation capability of a novel miniaturized capillary electrophoresis with an amperometric detection (μCE-AD) system was demonstrated by determining clenbuterol and salbutamol in real samples.The effects of several factors such as the acidity and concentration of the running buffer,the separation voltage,the applied potential and the injection time on CE-AD were examined and optimized.Under the optimum conditions,the two β-agonists could be baseline separated within 60 s at a separation voltage of 2 kV in a 90 mmol/L H3BO3-Na2B4O7 running buffer (pH 7.4),which was not interfered by ascorbic acid and uric acid.Highly linear response was obtained for above compounds over three orders of magnitude with detection limits ranging from 1.20 × 10-7 to 6.50× 10-8 mol/L (S/N=3).This method was successfully used in the analysis of feed and meat products with relatively simple extraction procedures.

  13. Simultaneous determination of vigabatrin and amino acid neurotransmitters in brain microdialysates by capillary electrophoresis with laser-induced fluorescence detection.

    Science.gov (United States)

    Benturquia, Nadia; Parrot, Sandrine; Sauvinet, Valérie; Renaud, Bernard; Denoroy, Luc

    2004-07-01

    Capillary electrophoresis with laser-induced fluorescence detection (CE-LIFD) coupled to in vivo microdialysis sampling was used in order to monitor simultaneously a drug and several neurotransmitters in the brain extracellular fluid. Determination of the antiepileptic drug vigabatrin and the amino acid neurotransmitters glutamate (Glu), l-aspartate (l-Asp) and gamma-aminobutyric acid (GABA) was performed on low-concentration samples which were derivatized with naphthalene-2,3-dicarboxaldehyde (NDA) and separated using a pH 9.2 75 mM sodium borate running buffer containing 60 mM sodium dodecyl sulfate (SDS) and 5mM hydroxypropyl-beta-cyclodextrin (HP-beta-CD). Glu, l-Asp and vigabatrin derivatized at a concentration of 1.0 x 10(-9) M, and GABA derivatized at a concentration of 5.0 x 10(-9) M, produced peaks with signal-to-noise ratios of 8:1, 8:1, 4:1 and 5:1, respectively. The nature of the neurotransmitter peaks found in rat brain microdialysates was confirmed by both electrophoretic and pharmacological validations. This method was used for monitoring vigabatrin and amino acid neurotransmitters in microdialysates from the rat striatum during intracerebral infusion of the drug and revealed rapid vigabatrin-induced changes in GABA and Glu levels. This original application of CE-LIFD coupled to microdialysis represents a powerful tool for pharmacokinetic/pharmacodynamic investigations.

  14. Cationic polyelectrolyte functionalized magnetic particles assisted highly sensitive pathogens detection in combination with polymerase chain reaction and capillary electrophoresis.

    Science.gov (United States)

    Chen, Jia; Lin, Yuexin; Wang, Yu; Jia, Li

    2015-06-01

    Pathogenic bacteria cause significant morbidity and mortality to humans. There is a pressing need to establish a simple and reliable method to detect them. Herein, we show that magnetic particles (MPs) can be functionalized by poly(diallyl dimethylammonium chloride) (PDDA), and the particles (PDDA-MPs) can be utilized as adsorbents for capture of pathogenic bacteria from aqueous solution based on electrostatic interaction. The as-prepared PDDA-MPs were characterized by Fourier-transform infrared spectroscopy, zeta potential, vibrating sample magnetometry, X-ray diffraction spectrometry, scanning electron microscopy, and transmission electron microscopy. The adsorption equilibrium time can be achieved in 3min. According to the Langmuir adsorption isotherm, the maximum adsorption capacities for E. coli O157:H7 (Gram-negative bacteria) and L. monocytogenes (Gram-positive bacteria) were calculated to be 1.8×10(9) and 3.1×10(9)cfumg(-1), respectively. The bacteria in spiked mineral water (1000mL) can be completely captured when applying 50mg of PDDA-MPs and an adsorption time of 5min. In addition, PDDA-MPs-based magnetic separation method in combination with polymerase chain reaction and capillary electrophoresis allows for rapid detection of 10(1)cfumL(-1) bacteria.

  15. Automated high-throughput dense matrix protein folding screen using a liquid handling robot combined with microfluidic capillary electrophoresis.

    Science.gov (United States)

    An, Philip; Winters, Dwight; Walker, Kenneth W

    2016-04-01

    Modern molecular genetics technology has made it possible to swiftly sequence, clone and mass-produce recombinant DNA for the purpose of expressing heterologous genes of interest; however, recombinant protein production systems have struggled to keep pace. Mammalian expression systems are typically favored for their ability to produce and secrete proteins in their native state, but bacterial systems benefit from rapid cell line development and robust growth. The primary drawback to prokaryotic expression systems are that recombinant proteins are generally not secreted at high levels or correctly folded, and are often insoluble, necessitating post-expression protein folding to obtain the active product. In order to harness the advantages of prokaryotic expression, high-throughput methods for executing protein folding screens and the subsequent analytics to identify lead conditions are required. Both of these tasks can be accomplished using a Biomek 3000 liquid handling robot to prepare the folding screen and to subsequently prepare the reactions for assessment using Caliper microfluidic capillary electrophoresis. By augmenting a protein folding screen with automation, the primary disadvantage of Escherichia coli expression has been mitigated, namely the labor intensive identification of the required protein folding conditions. Furthermore, a rigorous, quantitative method for identifying optimal protein folding buffer aids in the rapid development of an optimal production process. PMID:26678961

  16. Clinical application of a rapid microbiological test based on capillary zone electrophoresis to assess local skin infection

    Directory of Open Access Journals (Sweden)

    Szeliga Jacek

    2011-10-01

    Full Text Available Abstract Background The basic clinical problem associated with infection treatment is the fact that classic, commonly and routinely used isolation and identification methods are based on long-term processes of a phenotypic analysis of microorganisms. Consequently sometimes, especially in small centres, rapid implementation of antibacterial treatment becomes delayed. The work presents the initial results of rapid microbiological identification based on an original method of capillary zone electrophoresis (CZE. The study involved the analysis of 78 biological samples from post-operative wounds and trophic ulcers. Results The attempt was made to identify individual bacterial species based on characteristic features of electropherograms achieved. Finally, G(+ cocci type bacteria and different G(- rods were identified with sensitivity of 88.1% and specificity of 100%. Conclusions Based on the clinical trials using an electrophoretic technique in the field of microbiological diagnostics of infected exudate from a post-operative wound it can be concluded that it is a rapid and relatively sensitive method for initial identification of infectious pathogens.

  17. Rapid and sensitive quantification of C3- and C6-phosphoesters in starch by fluorescence-assisted capillary electrophoresis.

    Science.gov (United States)

    Verbeke, Jeremy; Penverne, Christophe; D'Hulst, Christophe; Rolando, Christian; Szydlowski, Nicolas

    2016-11-01

    Phosphate groups are naturally present in starch at C3- or C6-position of the glucose residues and impact the structure of starch granules. Their precise quantification is necessary for understanding starch physicochemical properties and metabolism. Nevertheless, reliable quantification of Glc-3-P remains laborious and time consuming. Here we describe a capillary electrophoresis method for simultaneous measurement of both Glc-6-P and Glc-3-P after acid hydrolysis of starch. The sensitivity threshold was estimated at the fg scale, which is compatible with the analysis of less than a μg of sample. The method was validated by analyzing antisense potato lines deficient in SBEs, GWD or GBSS. We show that Glc-3-P content is altered in the latter and that these variations do not correlate with modifications in Glc-6-P content. We anticipate the method reported here to be an efficient tool for high throughput study of starch phosphorylation at both C3- and C6-position. PMID:27516330

  18. Actinide oxalate complexes formation as a function of temperature by capillary electrophoresis coupled with inductively coupled plasma mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Brunel, Benoit; Mendes, Mickael; Aupiais, Jean [CEA, DAM, DIF, Arpajon (France); Philippini, Violaine [Nice Univ. Sophia Antipolis (France). Inst. de Chimie de Nice

    2015-05-01

    Complexation of various actinides (U(VI), Np(V), Pu(V), Am(III)) by oxalato ligand was studied by capillary electrophoresis (ICPMS detection) in 0.1 mol L{sup -1} NaClO{sub 4} ionic strength solutions at various temperatures (15, 25, 35, 45 and 55 C). For each solution a unique peak was observed as a result of a fast equilibrium between the free ions and the complexes (labile systems). The results confirmed the formation of the 1:1, 1:2 and 1:3 complexes for U(VI) and Am(III); the formation of the 1:1 and 1:2 complexes for Np(V) and the formation of only 1 complex for Pu(V). For each complex, the thermodynamic parameters (the Gibbs energy Δ{sub r}G(T), the molar entropy change Δ{sub r}S(T) and the molar enthalpy change Δ{sub r}H(T{sup 0})) were fitted to the experimental data. The effect of the ionic medium was treated using the specific ion interaction theory and the thermodynamic parameters at zero ionic strength were compared to previously published data.

  19. Capillary electrophoresis analysis of conventional splicing assays: IARC analytical and clinical classification of 31 BRCA2 genetic variants.

    Science.gov (United States)

    de Garibay, Gorka Ruiz; Acedo, Alberto; García-Casado, Zaida; Gutiérrez-Enríquez, Sara; Tosar, Alicia; Romero, Atocha; Garre, Pilar; Llort, Gemma; Thomassen, Mads; Díez, Orland; Pérez-Segura, Pedro; Díaz-Rubio, Eduardo; Velasco, Eladio A; Caldés, Trinidad; de la Hoya, Miguel

    2014-01-01

    Rare sequence variants in "high-risk" disease genes, often referred as unclassified variants (UVs), pose a serious challenge to genetic testing. However, UVs resulting in splicing alterations can be readily assessed by in vitro assays. Unfortunately, analytical and clinical interpretation of these assays is often challenging. Here, we explore this issue by conducting splicing assays in 31 BRCA2 genetic variants. All variants were assessed by RT-PCR followed by capillary electrophoresis and direct sequencing. If assays did not produce clear-cut outputs (Class-2 or Class-5 according to analytical International Agency for Research on Cancer guidelines), we performed qPCR and/or minigene assays. The latter were performed with a new splicing vector (pSAD) developed by authors of the present manuscript (patent #P201231427 CSIC). We have identified three clinically relevant Class-5 variants (c.682-2A>G, c.7617+1G>A, and c.8954-5A>G), and 27 analytical Class-2 variants (not inducing splicing alterations). In addition, we demonstrate that rs9534262 (c.7806-14T>C) is a BRCA2 splicing quantitative trait locus.

  20. Simultaneous analysis of Cu and Pb as ABEDTA complexes in Rhizoma coptidis by capillary electrophoresis coupled with solid phase extraction

    Institute of Scientific and Technical Information of China (English)

    Fan YunMeng; Ying Qin Wei; Heng Lu; Xin Xin Liu; Jin Xin Liu

    2012-01-01

    A novel capillary electrophoresis method for simultaneous determination of Cu and Pb has been developed in this paper.Cu(Ⅱ)and Pb(Ⅱ) ions were reacted with ABEDTA to form complex to achieve an ideal sensitivity of heavy metal complexes.The digestion solution of Rhizoma coptidis drug sample was purified by neutral Al2O3 column chromatography and the chromatographic behavior of metal-L complexes was investigated.The calibration curve was linear in the range of 5-60 μg/mL for Cu2+ and 5-25 μg/mL for Pb2+ with the correlation coefficients 0.9970 and 0.9972 for each (n =5).The average recoveries were 86.2% for Pb and 90.1% for Cu,while the relative standard deviations were 5.1% and 3.6% respectively.The method was successfully applied to determine Cu and Pb in R.coptidis drug samples.

  1. Development and validation of a capillary electrophoresis method for the determination of codeine, diphenhydramine, ephedrine and noscapine in pharmaceuticals.

    Science.gov (United States)

    Gomez, María R; Sombra, Lorena; Olsina, Roberto A; Martínez, Luis D; Silva, María F

    2005-01-01

    The present work describes a simple, accurate and rapid method for the separation and simultaneous determination of codeine, diphenhydramine, ephedrine and noscapine present in cough-cold syrup formulations by capillary zone electrophoresis. Factors affecting the separation were the buffer pH and concentration, applied voltage, and presence of additives. Separations were carried out in less than 10 min with a 20 mM sodium tetraborate buffer, pH 8.50. The carrier electrolyte gave baseline separation with good resolution, great reproducibility and accuracy. Calibration plots were linear over at least three orders of magnitude of analyte concentrations, the lower limits of detection being within the range 0.42-1.33 microg ml(-1). Detection was performed by UV absorbance at wavelengths of 205 and 250 nm. Quantification of the components in actual syrup formulations was calculated against the responses of freshly prepared external standard solutions. The method was validated and met all analysis requirements of quality assurance and quality control. The procedure was fast and reliable and commercial pharmaceuticals could be analyzed without prior sample clean-up procedure.

  2. An optimized capillary electrophoresis method for the simultaneous analysis of biomass degradation products in ionic liquid containing samples.

    Science.gov (United States)

    Aid, Tiina; Paist, Loore; Lopp, Margus; Kaljurand, Mihkel; Vaher, Merike

    2016-05-20

    An indirect capillary electrophoresis method for a quantitative determination of mono-, di- and oligosaccharides was developed to investigate biomass degradation, the isomerization of glucose into fructose and conversion of fructose to 5-hydroxymethylfurfural (5-HMF) in ionic liquids (ILs). Three chromophores, namely 2,6-pyridinedicarboxylic acid (PDC), maleic acid and phthalic acid, were used to perform indirect detection. The electroosmotic flow (EOF) was reversed to reduce analysis time, using 1-tetradecyl-3-methylimidazolium chloride (C14MImCl). The simultaneous separation of the underivatized mono-, di- and oligosaccharides was performed using four cellodextrin oligomers (cellotriose, cellotetraose, cellopentaose, cellohexaose), eight carbohydrates (xylose, fructose, glucose, galactose, lactose, cellobiose, raffinose, sucrose), two organic acids (acetic acid, levulinic acid) and 5-HMF. The best performance was obtained using background electrolyte (BGE) composed of 138.2mM NaOH, 40mM maleic acid and 5mMC14MImCl, the applied voltage was -21.7kV. The linear ranges for analyzed compounds were following: organic acids, raffinose and sucrose from 0.20 to 7mM, cellodextrin oligomers from 0.25 to 5mM, other analyzed carbohydrates from 0.25 to 7mM and 5-HMF from 0.05 to 7mM. The relative standard deviations (RSD) of peak areas varied from 3.47 to 9.62% during a 5-day analysis period and 0.58-5.29% during one day. PMID:27095128

  3. Stacking and Analysis of Melamine in Milk Products with Acetonitrile-Salt Stacking Technique in Capillary Electrophoresis

    Directory of Open Access Journals (Sweden)

    Yu Kong

    2014-01-01

    Full Text Available Melamine was measured in real milk products with capillary electrophoresis (CE based on acetonitrile-salt stacking (ASS method. Real milk samples were deproteinized with acetonitrile at a final concentration of 60% (v/v and then injected hydrodynamically at 50 mBar for 40.0 s. The optimized buffer contains 80.0 mmol/L pH 2.8 phosphates. Melamine could be detected within 20.0 min at +10 kV with a low limit of detection (LOD of 0.03 μmol/L. Satisfactory reproducibility (inter- and intraday RSD% both for migration time and peak area was lower than 5.0% and a wide linearity range of 0.05 μmol/L ~ 10.0 μmol/L were achieved. The proposed method was suitable for routine assay of MEL in real milk samples that was subjected to a simple treatment step.

  4. Separation of acidic and basic proteins by capillary electrophoresis using gemini surfactants and gemini-capped nanoparticles as buffer additives

    Institute of Scientific and Technical Information of China (English)

    LIU Qian; LI YanQing; YANG YanMin; YAO ShouZhuo

    2009-01-01

    This paper demonstrated simultaneous separation of acidic and basic proteins using cationic gemini surfactants as buffer additives in capillary electrophoresis. We showed that even at a low concentration (0.1 mmol·L~(-1)) of alkanediyl-α,ω-bis(dimethyloctadecylammonium bromide) (18-s-18), the wall adsorp-tion of both acidic and basic proteins could be effectively suppressed under acidic conditions. Smaller micelle size (e.g., s=5-8) is more effective for the separation of acidic proteins than larger micelle size (e.g., s 10). Varying the spacer length of gemini surfactants can influence the electrophoretic mobility and selectivity of proteins to achieve the desired separation. Under the optimized conditions, RSDs of the migration time were less than 0.8% and 2.2% for run-to-run and day-to-day assays, re-spectively, and protein recoveries ranged from 79% to 100.4%. Furthermore, we also investigated the use of gemini surfactant-capped gold nanoparticles (gemini@AuNPs) as buffer additives in protein separation. Introduction of AuNPs into the buffer shortened the analysis time and slightly improved the separation efficiencies. Finally, we presented the applications of this method in the analysis of bio-logical samples, including plasma, red blood cells and egg white.

  5. Separation of acidic and basic proteins by capillary electrophoresis using gemini surfactants and gemini-capped nanoparticles as buffer additives

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    This paper demonstrated simultaneous separation of acidic and basic proteins using cationic gemini surfactants as buffer additives in capillary electrophoresis. We showed that even at a low concentration (0.1 mmol·L-1) of alkanediyl-α,ω-bis(dimethyloctadecylammonium bromide) (18-s-18), the wall adsorption of both acidic and basic proteins could be effectively suppressed under acidic conditions. Smaller micelle size (e.g., s=5-8) is more effective for the separation of acidic proteins than larger micelle size (e.g., s<4 or >10). Varying the spacer length of gemini surfactants can influence the electrophoretic mobility and selectivity of proteins to achieve the desired separation. Under the optimized conditions, RSDs of the migration time were less than 0.8% and 2.2% for run-to-run and day-to-day assays, respectively, and protein recoveries ranged from 79% to 100.4%. Furthermore, we also investigated the use of gemini surfactant-capped gold nanoparticles (gemini@AuNPs) as buffer additives in protein separation. Introduction of AuNPs into the buffer shortened the analysis time and slightly improved the separation efficiencies. Finally, we presented the applications of this method in the analysis of bio-logical samples, including plasma, red blood cells and egg white.

  6. Establishment and application of a new method for the determination of kinetic parameters by plug-plug kinetic capillary electrophoresis (ppKCE)

    Institute of Scientific and Technical Information of China (English)

    XIONG CaiQiao; XIA ZhiNing; HUANG Rui; CHEN Hua; XU Pan

    2008-01-01

    A novel plug-plug kinetic capillary electrophoresis method was established, which can be used to si-multaneously determine the kinetic parameters kon and koff in interaction systems. The method is comparatively simple and some restrictions in conventional ppKCE methods can be effectively avoided, The requirements for resolution and detection sensitivity in this method are much lower than those of conventional ppKCE, The successful determination of the kinetic parameters and the binding constant Kb between citalopram and BSA showed availability of this method, The results were confirmed by us-ing the time ratio method. The application field of kinetic capillary electrophoresis can be expanded with this new method,

  7. Establishment and application of a new method for the determination of kinetic parameters by plug-plug kinetic capillary electrophoresis (ppKCE)

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    A novel plug-plug kinetic capillary electrophoresis method was established, which can be used to si-multaneously determine the kinetic parameters kon and koff in interaction systems. The method is comparatively simple and some restrictions in conventional ppKCE methods can be effectively avoided. The requirements for resolution and detection sensitivity in this method are much lower than those of conventional ppKCE. The successful determination of the kinetic parameters and the binding constant Kb between citalopram and BSA showed availability of this method. The results were confirmed by us-ing the time ratio method. The application field of kinetic capillary electrophoresis can be expanded with this new method.

  8. An interlaboratory comparison of ITS2-PCR for the identification of yeasts, using the ABI Prism 310 and CEQ8000 capillary electrophoresis systems

    Directory of Open Access Journals (Sweden)

    Verschraegen Gerda

    2005-03-01

    Full Text Available Abstract Background Currently, most laboratories identify yeasts routinely on the basis of morphology and biochemical reactivity. This approach has quite often limited discriminatory power and may require long incubation periods. Due to the increase of fungal infections and due to specific antifungal resistence patterns for different species, accurate and rapid identification has become more important. Several molecular techniques have been described for fast and reliable identification of yeast isolates, but interlaboratory exchangeability of identification schemes of molecular techniques has hardly been studied. Here, we compared amplified ITS2 fragment length determination by an ABI Prism 310 (Applied Biosystems, Foster City, Ca. capillary electrophoresis system with that obtained by a CEQ8000 (Beckman Coulter, Fullerton, Ca. capillary electrophoresis system. Results Although ITS2 size estimations on both systems differed and separate libraries had to be constructed for each system, both approaches had the same discriminatory power with regard to the 44 reference strains, identical identifications were obtained for 39/ 40 clinical isolates in both laboratories and strains from 51 samples were correctly identified using CEQ8000, when compared to phenotypic identification. Conclusion Identification of yeasts with ITS2-PCR followed by fragment analysis can be carried out on different capillary electrophoresis systems with comparable discriminatory power.

  9. Capillary Electrophoresis for the Simultaneous Determination of Metals by Using Ethylenediamine Tetraacetic Acid as Complexing Agent and Vancomycin as Complex Selector

    Institute of Scientific and Technical Information of China (English)

    THREEPROM, Jirasak; SOM-AUM, Waraporn; LIN, Jin-Ming

    2006-01-01

    A new separation system of capillary electrophoresis for the simultaneous determination of metals by using ethylenediamine tetraacetic acid (EDTA) as complexing agent and employing vancomycin as complex selector was described. The Z-shape cell capillary electrophoresis was used to enhance the sensitivity for the determination of the complexes of Cu(Ⅱ), Ni(Ⅱ), Co(Ⅱ) and Fe(Ⅲ) with EDTA. The partial filling method (co-current mode) was used in order to increase the selectivity of the electrophoretic method, meanwhile vancomycin was not present at the detector path during the detection of metal-EDTA complexes. The vancomycin concentration, phosphate concentration and pH of the buffer strongly influenced mobility, resolution and selectivity of the studied analytes. Under the optimal condition, the relative standard deviations (n=5) of the migration time and the peak area were less than 3.14% and 7.35%, respectively. Application of the Z-shape cell capillary electrophoresis method with UV detection and vancomycin loading led to the reliable determination of these metal ions in tap water and the recoveries were 97%-101%. The detection limits based on a signal to noise ratio of 3: 1 were found in the range of 2-10 μg·L-1.

  10. Quantitative analysis of pungent and anti-inflammatory phenolic compounds in olive oil by capillary electrophoresis.

    Science.gov (United States)

    Vulcano, Isabella; Halabalaki, Maria; Skaltsounis, Leandros; Ganzera, Markus

    2015-02-15

    The first CE procedure for the quantitative determination of pharmacologically relevant secoiridoids in olive oil, oleocanthal and oleacein, is described. Together with their precursors tyrosol and hydroxytyrosol they could be baseline separated in less than 15min using a borax buffer with pH 9.5, at 25kV and 30°C. Method validation confirmed that the procedure is selective, accurate (recovery rates from 94.0 to 104.6%), reproducible (σmax⩽6.8%) and precise (inter-day precision⩽6.4%), and that the compounds do not degrade quickly if non-aqueous acetonitrile is used as solvent. Quantitative results indicated a low occurrence of oleocanthal (0.004-0.021%) and oleacein (0.002-0.048%) in olive oil samples, which is in agreement to published HPLC data. The CE method impresses with its simple instrumental and methodological design, combined with reproducible and valid quantitative results. PMID:25236241

  11. Two phase capillary pumped heat transfer in the Instrument Thermal Test Bed

    Science.gov (United States)

    Didion, Jeffrey R.; Martins, Mario S.

    1992-01-01

    An experimental study of the thermal performance of two evaporators installed in the Instrument Thermal Test Bed (ITTB) was conducted. The ITTB was operated as a capillary pumped loop (CPL) with a transport length of approximately 12 meters. Empirical determinations of a general start up procedure, overall heat transfer coefficient, and minimum operating power were accomplished for each evaporator. Additionally, a detailed thermal model was developed for the High Power Spacecraft Thermal Management (HPSTM) evaporator and validated.

  12. Sensitive redox speciation of iron, neptunium, and plutonium by capillary electrophoresis hyphenated to inductively coupled plasma sector field mass spectrometry.

    Science.gov (United States)

    Graser, Carl-Heinrich; Banik, Nidhu Lal; Bender, Kerstin Anne; Lagos, Markus; Marquardt, Christian Michael; Marsac, Rémi; Montoya, Vanessa; Geckeis, Horst

    2015-10-01

    The long-term safety assessment for nuclear waste repositories requires a detailed understanding of actinide (geo)chemistry. Advanced analytical tools are required to gain insight into actinide speciation in a given system. The geochemical conditions in the vicinity of a nuclear repository control the redox state of radionuclides, which in turn has a strong impact on their mobility. Besides the long-lived radionuclides plutonium (Pu) and neptunium (Np), which are key elements in high level nuclear waste, iron (Fe) represents a main component in natural systems controlling redox-related geochemical processes. Measuring the oxidation state distribution for redox sensitive radionuclides and other metal ions is challenging at trace concentrations below the detection limit of most available spectroscopic methods (≥10(-6) M). Consequently, ultrasensitive new analytical techniques are required. Capillary electrophoresis (CE) is a suitable separation method for metal cations. CE hyphenated to inductively coupled plasma sector field mass spectrometry (CE-ICP-SF-MS) was used to measure the redox speciation of Pu (III, IV, V, VI), Np (IV, V, VI), and Fe (II, III) at concentrations lower than 10(-7) M. CE coupling and separation parameters such as sample gas pressure, make up flow rate, capillary position, auxiliary gas flow, as well as the electrolyte system were optimized to obtain the maximum sensitivity. We obtain detection limits of 10(-12) M for Np and Pu. The various oxidation state species of Pu and Np in different samples were separated by application of an acetate-based electrolyte system. The separation of Fe (II) and Fe (III) was investigated using different organic complexing ligands, EDTA, and o-phenanthroline. For the Fe redox system, a limit of detection of 10(-8) M was calculated. By applying this analytical system to sorption studies, we were able to underline previously published results for the sorption behavior of Np in highly diluted concentrations, and

  13. Studies on the structural variations of pentosan polysulfate sodium (NaPPS) from different sources by capillary electrophoresis.

    Science.gov (United States)

    Degenhardt, M; Ghosh, P; Wätzig, H

    2001-01-01

    Commercial pentosan polysulfate sodium salts (NaPPS) are highly sulfated polysaccharides derived from beechwood hemicellulose by sulfate esterification with a Mrel range of 1500-6000. The polysaccharide backbone of NaPPS consists of repeating linear units of 1-4 linked beta-D-xylopyranose with laterally substituted 4-methylglucopyranosyluronic acid units glycosidically linked to the 2 position of the main chain at every 10th xylopyranose unit on average. For many years NaPPS has been used for antithrombotic prophylaxis in Europe and interstitial cystitis in the USA and Australia. More recently NaPPS has found veterinary application for the treatment of osteoarthritis and related conditions in domestic animals and is registered for this use in Australia, New Zealand, Canada, UK, Eire, and several Scandanavian countries. At present the use of NaPPS for human disorders is confined to material manufactured by one company. However, for veterinary applications, NaPPS from three manufactures have been described. Since it is well known that the biological activities of sulfated polysaccharides are dependent on their molecular structures we considered it important to characterise these various NaPPS preparations using an established method of analysis. Unfortunately, traditional analytical techniques such as TLC, OR, UV/Vis spectroscopy, and size exclusion chromatography were incapable of providing structural information which would distinguish these NaPPS preparations from each other. In contrast, a capillary zone electrophoresis (CZE) method facilitated characterisation of the different NaPPS by a highly reproducible fingerprint, using a benzene-1,2,4-tricarboxylic acid buffer (8.75 mmol/L, pH = 4.9) with indirect UV detection (lambda = 217 nm) and a special capillary pre-treatment (1 M NaOH for 1 h at 25 degrees C, then running buffer for 120 min at 25 degrees C applying -20 kV). In the present study more than 20 batches of NaPPS from the three manufacturers have been

  14. Sensitive redox speciation of iron, neptunium, and plutonium by capillary electrophoresis hyphenated to inductively coupled plasma sector field mass spectrometry.

    Science.gov (United States)

    Graser, Carl-Heinrich; Banik, Nidhu Lal; Bender, Kerstin Anne; Lagos, Markus; Marquardt, Christian Michael; Marsac, Rémi; Montoya, Vanessa; Geckeis, Horst

    2015-10-01

    The long-term safety assessment for nuclear waste repositories requires a detailed understanding of actinide (geo)chemistry. Advanced analytical tools are required to gain insight into actinide speciation in a given system. The geochemical conditions in the vicinity of a nuclear repository control the redox state of radionuclides, which in turn has a strong impact on their mobility. Besides the long-lived radionuclides plutonium (Pu) and neptunium (Np), which are key elements in high level nuclear waste, iron (Fe) represents a main component in natural systems controlling redox-related geochemical processes. Measuring the oxidation state distribution for redox sensitive radionuclides and other metal ions is challenging at trace concentrations below the detection limit of most available spectroscopic methods (≥10(-6) M). Consequently, ultrasensitive new analytical techniques are required. Capillary electrophoresis (CE) is a suitable separation method for metal cations. CE hyphenated to inductively coupled plasma sector field mass spectrometry (CE-ICP-SF-MS) was used to measure the redox speciation of Pu (III, IV, V, VI), Np (IV, V, VI), and Fe (II, III) at concentrations lower than 10(-7) M. CE coupling and separation parameters such as sample gas pressure, make up flow rate, capillary position, auxiliary gas flow, as well as the electrolyte system were optimized to obtain the maximum sensitivity. We obtain detection limits of 10(-12) M for Np and Pu. The various oxidation state species of Pu and Np in different samples were separated by application of an acetate-based electrolyte system. The separation of Fe (II) and Fe (III) was investigated using different organic complexing ligands, EDTA, and o-phenanthroline. For the Fe redox system, a limit of detection of 10(-8) M was calculated. By applying this analytical system to sorption studies, we were able to underline previously published results for the sorption behavior of Np in highly diluted concentrations, and

  15. Analysis of thiols by microchip capillary electrophoresis for in situ planetary investigations.

    Science.gov (United States)

    Mora, Maria F; Stockton, Amanda M; Willis, Peter A

    2013-01-01

    The detection of thiols on extraterrestrial bodies could provide evidence for life, as well as a host of potential prebiological or abiological processes. Here, we report a novel protocol to analyze organic thiols by microchip CE with LIF detection. Thiols were labeled with Pacific Blue C5 maleimide and analyzed by MEKC. The separation buffer consisted of 15 mM tetraborate pH 9.2 and 25 mM SDS. The optimized method provided LODs ranging from 1.4 to 15 nM. The method was validated using samples collected from geothermal pools at Hot Creek Gorge, California, which were found to contain 2-propanethiol and 1-butanethiol in the nanomolar concentration range. These samples serve as chemical analogues to material potentially present in the reducing environment of primitive Earth and also at sulfurous regions of Mars. Hence, the protocol developed here enables highly sensitive thiol analysis in samples with complexity comparable to that expected in astrobiologically relevant extraterrestrial settings. This new protocol could be readily added to the existing suite of microfluidic chemical analyses developed for in situ planetary exploration; all that is required is the incorporation of two new reagents to the payload of an existing instrument concept. PMID:23161601

  16. Quantum dot-DNA aptamer conjugates coupled with capillary electrophoresis: A universal strategy for ratiometric detection of organophosphorus pesticides.

    Science.gov (United States)

    Tang, Tingting; Deng, Jingjing; Zhang, Min; Shi, Guoyue; Zhou, Tianshu

    2016-01-01

    Based on the highly sensitivity and stable-fluorescence of water-soluble CdTe/CdS core-shell quantum dots (QDs) with broad-specificity DNA aptamers, a novel ratiometric detection strategy was proposed for the sensitive detection of organophosphorus pesticides by capillary electrophoresis with laser-induced fluorescence (CE-LIF). The as-prepared QDs were first conjugated with the amino-modified oligonucleotide (AMO) by amidation reaction, which is partial complementary to the DNA aptamer of organophosphorus pesticides. Then QD-labeled AMO (QD-AMO) was incubated with the DNA aptamer to form QD-AMO-aptamer duplex. When the target organophosphorus pesticides were added, they could specifically bind the DNA aptamer, leading to the cleavage of QD-AMO-aptamer duplex, accompany with the release of QD-AMO. As a result, the ratio of peak height between QD-AMO and QD-AMO-aptamer duplex changed in the detection process of CE-LIF. This strategy was subsequently applied for the detection of phorate, profenofos, isocarbophos, and omethoate with the detection limits of 0.20, 0.10, 0.17, and 0.23μM, respectively. This is the first report about using QDs as the signal indicators for organophosphorus pesticides detection based on broad-specificity DNA aptamers by CE-LIF, thus contributing to extend the scope of application of QDs in different fields. The proposed method has great potential to be a universal strategy for rapid detection of aptamer-specific small molecule targets by simply changing the types of aptamer sequences.

  17. Analysis of recombinant human erythropoietin glycopeptides by capillary electrophoresis electrospray-time of flight-mass spectrometry.

    Science.gov (United States)

    Giménez, Estela; Ramos-Hernan, Raquel; Benavente, Fernando; Barbosa, José; Sanz-Nebot, Victoria

    2012-01-01

    Capillary electrophoresis electrospray-mass spectrometry was used to detect and characterize the great variety of O- and N-glycopeptide glycoforms of recombinant human erythropoietin (rhEPO) using an orthogonal accelerating time-of-flight mass spectrometer to obtain their exact molecular masses (CE-TOF-MS). rhEPO was digested with trypsin and Glu-C and analyzed by CE-TOF-MS to detect O(126), N(83), N(24)-N(38) and N(24) and N(38) glycopeptide glycoforms, respectively. Neuraminidase was first used to enhance the detection of the glycopeptides and detect all possible glycoforms contained in each glycosylation site. O(126) and N(83) glycopeptides were extensively characterized. Twelve sialoforms corresponding to 5 different glycoforms were detected in N(83), and for the first time, a sulfated sialoform of this glycopeptide was also detected. In the case of O(126), different sialoforms with different types of sialic acids (Neu5Gc and Neu5Ac) were detected and an estimation of the relative percentage of Neu5Gc versus Neu5Ac was also carried out for this glycopeptide. N(24) and N(38) glycosylation sites were also characterized by CE-TOF-MS after Glu-C digestion and these results permitted to rule out some glycan combinations for N(24)-N(38) glycopeptide glycoforms. This study provided a reliable glycopeptide map of rhEPO and may be regarded as an excellent starting point to analyze rhEPO glycopeptides in biological fluids and detect the use of this hormone in sports. PMID:22122935

  18. Thermodynamics of binding of a sulfonamide inhibitor to metal-mutated carbonic anhydrase as studied by affinity capillary electrophoresis.

    Science.gov (United States)

    Sato, Yosuke; Hoshino, Hitoshi; Iki, Nobuhiko

    2015-09-01

    By affinity capillary electrophoresis (ACE), the thermodynamic binding constants of a sulfonamide (SA) inhibitor to bovine carbonic anhydrase II (CA) and metal mutated variants (M-CAs) were evaluated. 1-(4-Aminosulfonylphenylazo)-2-naphthol-6,8-disulfonate was used as the SA in the electrophoretic buffer for ACE. The Scatchard analysis of the dependence of the electrophoretic mobility of native CA on the SA concentration provided the binding constant to be Kb=(2.29±0.05)×10(6) M(-1) (at pH8.4, 25°C). On the other hand, apoCA showed far smaller value [Kb=(3.76±0.14)×10(2) M(-1)], suggesting that the coordination of SA to the Zn(II) center controlled the binding thermodynamics. The ACE of M-CAs showed the same behaviors as native CA but with different Kb values. For example, Co-CA adopting the same tetrahedral coordination geometry as native CA exhibited the largest Kb value [(2.55±0.05)×10(6) M(-1)] among the M-CAs. In contrast, Mn- and Ni-CA, which adopted the octahedral coordination geometry, had Kb values that were about two orders of magnitude lower. Because the hydrophobic cavity of CA around the active center pre-organized the orientation of SA, thereby fixing the ligating NH(-) moiety to the apex of the tetrahedron supported by three basal His3 of CA, metals such as Zn and Co at the center of M-CA gave the most stable CA-SA complex. However, pre-organization was not favored for octahedral geometry. Thus, pre-organization of SA was the key to facilitating the tetrahedral coordination geometry of the Zn(II) active center of CA.

  19. Improvement of Mitochondria Extract from Saccharomyces cerevisiae Characterization in Shotgun Proteomics Using Sheathless Capillary Electrophoresis Coupled to Tandem Mass Spectrometry.

    Science.gov (United States)

    Ibrahim, Marianne; Gahoual, Rabah; Enkler, Ludovic; Becker, Hubert Dominique; Chicher, Johana; Hammann, Philippe; François, Yannis-Nicolas; Kuhn, Lauriane; Leize-Wagner, Emmanuelle

    2016-04-01

    In this work, we describe the characterization of a quantity-limited sample (100 ng) of yeast mitochondria by shotgun bottom-up proteomics. Sample characterization was carried out by sheathless capillary electrophoresis, equipped with a high sensitivity porous tip and coupled to tandem mass spectrometry (CESI-MS-MS) and concomitantly with a state-of-art nano flow liquid chromatography coupled to a similar mass spectrometry (MS) system (nanoLC-MS-MS). With single injections, both nanoLC-MS-MS and CESI-MS-MS 60 min-long separation experiments allowed us to identify 271 proteins (976 unique peptides) and 300 proteins (1,765 unique peptides) respectively, demonstrating a significant specificity and complementarity in identification depending on the physicochemical separation employed. Such complementary, maximizing the number of analytes detected, presents a powerful tool to deepen a biological sample's proteomic characterization. A comprehensive study of the specificity provided by each separating technique was also performed using the different properties of the identified peptides: molecular weight, mass-to-charge ratio (m/z), isoelectric point (pI), sequence coverage or MS-MS spectral quality enabled to determine the contribution of each separation. For example, CESI-MS-MS enables to identify larger peptides and eases the detection of those having extreme pI without impairing spectral quality. The addition of peptides, and therefore proteins identified by both techniques allowed us to increase significantly the sequence coverages and then the confidence of characterization. In this study, we also demonstrated that the two yeast enolase isoenzymes were both characterized in the CESI-MS-MS data set. The observation of discriminant proteotypic peptides is facilitated when a high number of precursors with high-quality MS-MS spectra are generated.

  20. High-throughput molecular determination of salmonella enterica serovars by use of multiplex PCR and capillary electrophoresis analysis.

    Science.gov (United States)

    Leader, Brandon T; Frye, Jonathan G; Hu, Jinxin; Fedorka-Cray, Paula J; Boyle, David S

    2009-05-01

    Salmonella enterica is a leading cause of food-borne illness worldwide and is also a major cause of morbidity and mortality in domestic and wild animals. In the current study, a high-throughput molecular assay was developed to determine the most common clinical and nonhuman serovars of S. enterica in the United States. Sixteen genomic targets were identified based on their differential distribution among common serovars. Primers were designed to amplify regions of each of these targets in a single multiplex PCR while incorporating a 6-carboxyfluorescein-labeled universal primer to fluorescently label all amplicons. The fluorescently labeled PCR products were separated using capillary electrophoresis, and a Salmonella multiplex assay for rapid typing (SMART) code was generated for each isolate, based upon the presence or absence of PCR products generated from each target gene. Seven hundred fifty-one blind clinical isolates of Salmonella from Washington State, collected in 2007 and previously serotyped via antisera, were screened with the assay. A total of 89.6% of the isolates were correctly identified based on comparison to a panel of representative SMART codes previously determined for the top 50 most common serovars in the United States. Of the remaining isolates, 6.2% represented isolates that produced a new SMART code for a previously determined serotype, while the final 8.8% were from serotypes not screened in the original panel used to score isolates in the blinded study. This high-throughput multiplex PCR assay allowed simple and accurate typing of the most prevalent clinical serovars of Salmonella enterica at a level comparable to that of conventional serotyping, but at a fraction of both the cost and time required per test. PMID:19261787