Commercial Capillary Electrophoresis System Research Articles

Emerging techniques for the detection of pyrotechnic residues from seized postal packages containing fireworks

High volume screening of parcels with the aim to trace the illegal distribution and selling of fireworks using postal services is challenging. Inspection services have limited manpower and means to perform extensive visual inspection. In this study, the presence of solid pyrotechnic residues collected from cardboard shipping parcels containing fireworks was investigated for direct in-field chemical detection. Two emerging trace detection techniques, i.e., capillary electrophoresis (CE)-based inorganic oxidizer detector and infrared thermal desorption (IRTD) coupled with direct analysis in real time mass spectrometry (DART-MS), were investigated for their potential as screening tools. Detection of non-visible pyrotechnic trace residues from real-case seized parcels was demonstrated using both screening techniques. However, the high nitrate background in the commercial CE system complicated its screening for black powder traces. IRTD-DART-MS allowed differentiation between flash and black powder by identification of the molecular inorganic ions. Compared to the portable CE instrument, rapid screening using IRTD-DART-MS is currently limited to laboratory settings. The capabilities of these emerging techniques established solid particle and trace residue chemical detection as interesting options for parcel screening in a logistic setting.

A Miniature 3D Printed LED-Induced Fluorescence Detector for Capillary Electrophoresis and Dual-Detector Taylor Dispersion Analysis.

Taylor dispersion analysis (TDA) provides absolute determination of diffusion coefficients for analytes ranging from small molecules to particulate matter. TDA has seen a resurgence in recent years, as modern commercial capillary electrophoresis (CE) instrumentation is well equipped to meet the precision flow requirements of TDA. Discontinuous flow velocities, which occur during sample injection, can lead to substantial inaccuracies in single-point detection TDA. Dual-point detection allows TDA to be carried out under continuous flow in the volume between the detection points, but dual-point fluorescence detection has not previously been feasible within the confines of commercial CE instrumentation. Here, we describe a compact light-emitting diode (LED)-induced fluorescence detector designed for online, dual-point capillary detection within a commercial CE system. The three-dimensional (3D) printed detector houses an inexpensive LED excitation source, a bandpass excitation filter, an integral 3D printed pinhole collimator, and a ball lens, which collects fluorescence emission. Multivariate optimization of operating conditions yielded a detection limit of 613 ± 13 pM for CE of fluorescein disodium salt solution in borate buffer. The miniature size of the device allowed integration of two detectors within a commercial CE system without modification to the instrument, thereby enabling dual-detector assays including TDA and CE-TDA. Monitoring of the bioconjugation reaction between fluorescein isothiocyanate (FITC) and a model protein illustrates the utility of direct, calibration-free size determination, which enabled the resolution of fluorescence originating from free FITC from that of protein-bound FITC. TDA detection coupled to CE enabled the determination of peak identities without the need for standard solutions.

The Use of a 3D-Printed Microfluidic Device and Pressure Mobilization for Integrating Capillary Electrophoresis with Electrochemical Detection.

Capillary electrophoresis coupled with electrochemical detection can be a powerful analysis tool; however, previous methods developed to integrate these two techniques can often times be fragile and have alignment issues such that there are no commercially available approaches. In this paper, we present the use of a 3D-printed Wall-Jet Electrode device for integrating capillary electrophoresis with electrochemical detection. A pressure mobilization step was also utilized to further reduce noise by allowing the electrophoresis separation step to continue only until the first analyte was close to elution. Then, the separation voltage was terminated and pressure-based flow was used for elution of the analyte bands onto the electrode surface with a wall-jet configuration. It is shown that the pressure-based elution is beneficial for the reduction of baseline noise and elimination of field effects. A mixture of catecholamines were separated to demonstrate effectiveness of the system. In addition, the system was coupled with a Beckman Coulter commercial capillary electrophoresis instrument in a straightforward manner. The system was also shown to be effective in separations done with a high ionic strength physiological buffer. This 3D printing approach can be used by researchers to utilize electrochemical detection on commercial capillary electrophoresis systems by downloading the provided STL and/or CAD files.

Optimization of ELFSE DNA sequencing with EOF counterflow and microfluidics.

We present a nonlinear optimization study of different implementations of the DNA electrophoretic method "End-labeled Free-solution Electrophoresis" in commercial capillary electrophoresis systems and microfluidics to improve the time required for readout. Here, the effect of electro-osmotic counterflows and snap-shot detection are considered to allow for detection of peaks soon after they are electorphoretically resolved. Using drag tags available in micelle form, we identify a design capable of sequencing 600 bases in 2.8 min.

Comparative study between a commercial and a homemade capillary electrophoresis instrument for the simultaneous determination of aminated compounds by induced fluorescence detection

The performance of two capillary electrophoresis (CE) instruments, one commercial and one homemade device, were compared for the determination of derivatised aminated compounds with fluorescein isothiocyanate (FITC). The commercial CE system first uses an argon ion laser as excitation source; the homemade CE device uses an inexpensive blue-light-emitting diode (LED) as the light source and a charge-coupled device (CCD) as the detection system. After fine optimisation of several separation parameters in both devices, a co-electroosmotic flow CE methodology was achieved in coated capillary tubing with 0.001% hexadimetrine bromide (HDB), and 50 mmol L-1 sodium borate at pH 9.3 with 20% 2-propanol for the determination of several amines and aminoacids. Analytical performances, applicability in beer samples and other aspects such as cost or potential for miniaturization have been compared for both devices.

Miniaturized fluorescence detection chip for capillary electrophoresis immunoassay of agricultural herbicide atrazine

This paper reports a miniaturized fluorescence detection chip for capillary electrophoresis immunoassay of atrazine, which effectively reduces the size of fluorescence detection system. The photodiode with fluorescence filter was embedded in PDMS (polydimethylsiloxane) microfluidic chip and was placed just below the microfluidic channel. This detection chip is only few mm thick without loss of fluorescence due to the proximity of photodiode and channel. To investigate the feasibility of in situ detection of agricultural herbicide, atrazine was detected using capillary electrophoresis immunoassay in microfluidic chip. Mixture of 570 nM fluorescence-labeled atrazine (Ag*) and 700 nM anti-atrazine antibody (Ab) was injected and separated in 25 mm long microfluidic channel. The separated peaks of Ab-Ag* immunocomplex and Ag* were detected by the miniaturized detector and the change of peak magnitude was also observed with the variation of Ab concentration. The result was verified with those of external PMT (photomultiplier tube) and commercial capillary electrophoresis system. Hence, we have demonstrated the feasibility of portable CE immunoassay of atrazine with on-chip fluorescence detector.

Automated analysis of individual particles using a commercial capillary electrophoresis system

Capillary electrophoretic analysis of individual submicrometer size particles has been previously done using custom-built instruments. Despite that these instruments provide an excellent signal-to-noise ratio for individual particle detection, they are not capable of performing automated analyses of particles. Here we report the use of a commercial Beckman P/ACE MDQ capillary electrophoresis (CE) instrument with on-column laser-induced fluorescence (LIF) detection for the automated analysis of individual particles. The CE instrument was modified with an external I/O board that allowed for faster data acquisition rates (e.g. 100 Hz) than those available with the standard instrument settings (e.g. 4 Hz). A series of eight hydrodynamic injections expected to contain 32 +/- 6 particles, each followed by an electrophoretic separation at -300 V cm(-1) with data acquired at 100 Hz, showed 28 +/- 5 peaks corresponding to 31.9 particles as predicted by the statistical overlap theory. In contrast, a similar series of hydrodynamic injections followed by data acquisition at 4 Hz revealed only 8 +/- 3 peaks suggesting that the modified system is needed for individual particle analysis. Comparison of electropherograms obtained at both data acquisition rates also indicate: (i) similar migration time ranges; (ii) lower variation in the fluorescence intensity of individual peaks for 100 Hz; and (iii) a better signal-to-noise ratio for 4 Hz raw data. S/N improved for 100 Hz when data were smoothed with a binomial filter but did not reach the S/N values previously reported for post-column LIF detection. The proof-of-principle of automated analysis of individual particles using a commercially available CE system described here opens exciting possibilities for those interested in the study and analyses of organelles, liposomes, and nanoparticles.

Separation of fluorescent derivatives of hydroxyl-containing small molecules on a microfluidic chip

Capillary electrophoresis (CE) on a microchip has been employed in this work because of the need to separate and detect fluorescent derivatives on some licorice-derived compounds, namely glycyrrhizin (GL), isoliquiritigenin (IQ). These compounds, which contain only phenolic or aliphatic hydroxyl (secondary) groups, are hard to derivatize in aqueous solutions. Although there are some fluorescent reagents that will react with hydroxyl groups, these reactions are usually achieved in aprotic solvents, but not in aqueous solutions. The only fluorescent reagent that works in aqueous solvents is 5-([4,6-dichlorotriazin-2-yl]amino)fluorescein (DTAF), but it has mostly been employed to label macromolecules. For the first time, we have successfully fluorescently label two hydroxyl-containing small molecules, GL and IQ in aqueous solutions. The reaction products were separated by CE and detected fluorescently on a microfluidic chip within 100 s. For comparison, the separation was also carried out on a commercial CE system using UV absorbance detection.

Quantitative capillary zone electrophoresis of inorganic anions.

Despite the availability of commercial capillary electrophoresis systems for over ten years, where quantitative analysis is required, capillary zone electrophoresis (CZE) has often failed to replace ion chromatography as the method of choice for a large number of analytes, not least inorganic anions. To investigate the reasons for this apparent failing, a review is presented of work that has been carried out to-date involving the quantitative application of CZE to the determination of inorganic anions in industrial and environmental samples. This review summarizes work both investigating and improving the quantitative aspects of the CZE of inorganic anions. A complete survey of how CZE has been applied to the determination of inorganic anions in real samples is given, including what, if any, analytical performance parameters were investigated and quoted, and if quality assurance data and validation methods were briefly considered, thoroughly investigated or simply ignored.

Improving signal to background ratio for on-the-fly fluorescence lifetime detection in capillary electrophoresis.

On-the-fly fluorescence lifetime detection (OFLD) in capillary electrophoresis (CE) was previously demonstrated using a commercial multiharmonic Fourier transform (MHF) spectrofluorometer interfaced to a commercial CE system. This paper discusses optimization of the interface design for minimization of background fluorescence and scattered light, thereby maximizing the signal-to-background ratio (S/B) of the dynamic measurement. Strategies included using various combinations of optical filters including a holographic filter and longpass or bandpass filters, tilting the capillary relative to the incident laser beam, employing a confocal design and adding an iris to remove out-of-focus light, using a microscope objective in the emission beam to increase the collection efficiency, and using square instead of ciruclar capillary columns. Significant improvements in S/B for on-column, on-the-fly detection of fluorescein in CE were achieved with most modifications.

Capillary electrophoresis--time-of-flight mass spectrometry of drugs of abuse.

The use of capillary electrophoresis (CE) for the determination of drugs of abuse was explored. A commercial CE system was interfaced with a laboratory-built time-of-flight mass spectrometer (TOFMS) which was equipped with a high-speed data acquisition system to provide accurate monitoring of efficient separations. Ionization of the CE eluent was achieved with an electrospray ionization source. Standard mixtures and seized samples were analyzed either by direct infusion of the analyte solutions or after separation by CE. Detection at the low femtomole level was obtained using CE-TOFMS.

Automated microanalysis using magnetic beads with commercial capillary electrophoretic instrumentation

The potential of a new microanalytical method using magnetic beads (MBs) and commercial capillary electrophoresis (CE) instrumentation for performing enzymatic and inhibition assays, as well as for analysis of biological molecules such as antigens, substrates, etc., has been explored. A small quantity of magnetic beads containing immobilized biomolecules was injected into a neutral hydrophilic-coated fused-silica capillary. The short plug (2–3 mm) of beads was held fixed by a magnet placed in the cartridge of the CE system, without the use of frits. The beads could be replaced after each run, eliminating the need to regenerate the solid support. Two protocols were used for analysis: sequential injection (SI) and SI followed by isotachophoretic (ITP) focusing. Alkaline phosphatase (AP) and HIV-protease were used to demonstrate the SI procedure for enzymatic and inhibition assays. The second protocol, SI/ITP, was employed to quantitate an antigen (mouse mAB) using antibodies (sheep IgG towards mouse AB) immobilized on the beads. The MB-CE method, requiring only femtomole (fmol) quantities of material, can potentially be employed in diagnostic and forensic assays, kinetic studies and searching for inhibitors, ligands, receptors, etc.

Applicability of Capillary Electrophoresis with Amperometric Detection to Study Degradation of Chlorophenols in Contaminated Soil

Details are presented for assembling an amperometric detector compatible with commercial capillary electrophoresis (CE) instruments. The detector assembled with a gold electrode achieves a sufficient sensitivity in analyzing liquid extracts to enable determination of pentachlorophenol (PCP) in soils at 35 mg/kg (PCP/soil). The CE system equipped with amperometric detection is used to study the oxidation of chlorophenols with ceric sulfate. The selected chlorophenols are completely oxidized by ceric sulfate in 5 min and the ceric sulfate can be recovered and reused. The same CE system is also applied for monitoring the photodegradation of PCP in contaminated soil, in the presence of titanium dioxide. Complete degradation of PCP, with no trace of electroactive products, is achieved in 7 h. This study demonstrates that capillary electrophoresis can be an important technique for monitoring processes of degradation and bioremediation.

Simple double-beam absorption detection systems for capillary electrophoresis based on diode lasers and light-emitting diodes

Simple double-beam absorption detection systems for capillary electrophoresis (CE) based on diode lasers and light-emitting diodes (LEDs) are developed. Using digital normalization, the intensity fluctuations are greatly reduced and a relative standard deviation of the background noise of 10 −5 to 10 −6 can be achieved. The detection performance is thus improved over those of commercial CE systems. For the diode-laser-based system, the gain in detectability results from both a reduction in intensity fluctuations and a better optical coupling of the laser beam with the small capillary tube, maximizing the effective optical path length. The improvement in the LED-based system is largely due to the excellent stability of the LED augmented by double-beam cancellation of the baseline fluctuations. Indirect absorption detection for cationic and anionic species are also demonstrated, extending the applicability of the system.

Sensitive fluorescence detection in capillary electrophoresis using laser diodes and multiplex dyes

Abstract A detector for laser induced fluorescence based on laser diole technology was designed and sensitivity measurements with rhodamine 800 were performed. The detection limit is around 100 molecules in the detection volume. Using the laser diode in combination with a commercial capillary electrophoresis system, about 1200 molecules were detectable. Using time-resolved fluorescence measurements multiplex dyes were recognized with a pulsed laser diode and a pattern recognition technique collecting only a few hundred photons.

On-column double-beam laser absorption detection for capillary electrophoresis

Double-beam laser absorption detection in capillary electrophoresis (CE) has been developed. This is based on the direct subtraction of reference and signal photocurrents by an electronic circuit, under feedback control, to reduce background noise. A simple equation for calculating concentrations has been proposed and was confirmed by experimental results. A practical noise-to-signal ratio of 1 [times] 10[sup [minus]5] in intensity is achieved. This is 5 times lower than that of commercial CE systems. For absorbance detection, as low as 2 [times] 10[sup [minus]8] M malachite green can be detected. This corresponds to a 25-fold improvement of detection limit over commercial systems. This gain in detectability results from both a reduction in intensity fluctuations (noise) and an increase in the effective absorption path length (signal). 22 refs., 6 figs.

Comparison of liquid-junction and coaxial interfaces for capillary electrophoresis-mass spectrometry with application to compounds of concern to the aquaculture industry

The application of capillary electrophoresis-mass spectrometry (CE-MS) to the analysis of compounds of concern to the aquaculture industry is reported. Two different approaches to coupling the CE column to an IonSpray atmospheric pressure ionization (API) interface, viz., a liquid-junction and a coaxial arrangement, are described and compared with regard to ruggedness, ease of use, sensitivity and electrophoretic performance. The different injection modes used in three commercial capillary electrophoresis systems were also evaluated for their applicability to CE-MS. The use of CE-MS for the analysis of a variety of classes of antibiotics used in the fish aquaculture industry, such as the sulfonamides and their potentiators ( e.g., trimethoprim), is demonstrated and was used to confirm the presence of these components in shellfish extracts at the low ppm level. CE-MS was also applied to the analysis of marine toxins such as saxitoxin and its analogues which are associated with paralytic shellfish poisoning, and also to the toxins responsible for amnesic and diarrhetic shelfish poisoning. Tandem mass spectrometry (MS-MS) was used to provide structural information on these analytes, and the ability to distinguish isometric compounds based on their different migration and fragmentation characteristics using CE-MS-MS is demonstrated.

The nature and speed of flashpowders

High volume screening of parcels with the aim to trace the illegal distribution and selling of fireworks using postal services is challenging. Inspection services have limited manpower and means to perform extensive visual inspection. In this study, the presence of solid pyrotechnic residues collected from cardboard shipping parcels containing fireworks was investigated for direct in-field chemical detection. Two emerging trace detection techniques, i.e., capillary electrophoresis (CE)-based inorganic oxidizer detector and infrared thermal desorption (IRTD) coupled with direct analysis in real time mass spectrometry (DART-MS), were investigated for their potential as screening tools. Detection of non-visible pyrotechnic trace residues from real-case seized parcels was demonstrated using both screening techniques. However, the high nitrate background in the commercial CE system complicated its screening for black powder traces. IRTD-DART-MS allowed differentiation between flash and black powder by identification of the molecular inorganic ions. Compared to the portable CE instrument, rapid screening using IRTD-DART-MS is currently limited to laboratory settings. The capabilities of these emerging techniques established solid particle and trace residue chemical detection as interesting options for parcel screening in a logistic setting.