Electrokinetic Injection Mode Research Articles

Analysis of honey using capillary electrophoresis with electrokinetic injection – Direct analysis of viscous samples without sample pretreatment

Although the electrokinetic injection mode is applicable for direct introduction of ionic components from highly viscous samples, this is the first work which studies this special feature of capillary electrophoresis for this purpose. 9 metal cations were determined in honey utilizing direct injection, zone electrophoretic separation and indirect UV detection of the sample components. Reproducibility better than 4 RSD% was found for peak areas and the LOD values ranged between 0.045 and 0.44 mg/kg when injection parameters were set to 7.5 kV × 5 s. In order to remedy the quantitation problems arising from mobility and matrix bias, the applicability of the internal universal calibration was examined. It was found that in cases where the mobilities of cations do not differ from the mobilities of the monitoring ion and the internal universal standard (IUS) by more than 10%, the obtained results agreed well with the data measured with ICP-OES.

Quantitative on‐line concentration for capillary electrophoresis with inkjet sample introduction technique

A quantitative sample introduction method based upon inkjet injection was applied to capillary electrophoresis coupled with stacking and sweeping on-line concentration techniques. Methylxanthines were used as model compounds for the proof-of-concept of the method. The volume of injected sample could be easily manipulated by controlling the number of ejected droplets in the injection procedure. Under optimized conditions, a linear relationship between the ejected droplet number and peak area was obtained when the droplet number introduced into the capillary was less than 100. Under optimized quantitative on-line concentration conditions, the limits of detection for theobromine, caffeine, and theophylline were 1.0, 2.0, and 1.0 μM, respectively. The inkjet injection system was evaluated by comparing it with conventional injection methods. The electropherogram of the inkjet injection mode was the same as that for hydrodynamic injection mode, and no sample discrimination was observed compared with the electrokinetic injection mode. The established method was applied to the determination of methylxanthines in bottled green tea. The recoveries of theobromine, caffeine, and theophylline were 94.1, 110.6, and 86.8%, respectively. We conclude that proposed method can be used for quantitative concentration for capillary electrophoresis, thus resulting in an improved accuracy.

Analysis of buserelin in urine by online combination of capillary zone electrophoresis with electrospray mass spectrometry

A fast and precise analysis of the synthetic peptide buserelin in urine using CZE-ESI-MS method has been demonstrated. Formic acid at 50 mmol/L concentration served as background electrolyte in CZE stage and it is compatible with MS detection in positive ionization mode. Two injection modes were tested, i.e. pressure (50 mbar for 5 s) and electrokinetic injection (5 kV for 5 s), of which electrokinetic injection provided better calibration parameters. Buserelin LODs were 0.47 microg/mL in water and 0.63 microg/mL in ten times diluted urine samples using pressure injection, while they were 0.32 microg/mL in water and 0.34 microg/mL in ten times diluted urine samples using electrokinetic injection. Repeatability of buserelin migration times was below 6% (pressure injection mode) and 1% (electrokinetic injection mode). Repeatability of buserelin peak area in SIM mode (m/z=620.5+/-0.5) was less than 12% (pressure injection mode) and 5.8% (electrokinetic injection mode). In this work, no interferences were observed during the analyses of spiked urine samples.

Electrokinetic‐based injection modes for separative microsystems

Miniaturization of analytical instruments has attracted a wide interest in Analytical Chemistry over the past decade because of the advantages such as reduced reagent consumption and shorter analysis time. For chips involving separation, injection is a key step to achieve efficient and sensitive analysis. Electrokinetic injection mode is mostly used in chips because it is easier to generate flow motion in microsystems via electric potential control at channel extremities than pressure-driven flow. The injection step usually involves several intersecting channels. For each injection design, different injection modes can be done, depending on electric field sequences and distributions. This paper is an up-to-date review of these different modes on a chip.

Comparative study of floating and dynamic injection modes in electrokinetic separative microsystems

Miniaturization of analytical instruments has attracted a wide interest in analytical chemistry over the past decade because of the advantages of reduced reagent consumption, better analytical performance, and shorter analysis time. The widespread interest in this field has resulted in efforts to develop chips. For chips involving separation, injection is a key step to achieve efficient and sensitive analysis. This work presents a comparative study of two electrokinetic injection modes in chips: the floating, which has been mainly used up to now, and the dynamic. This study was done with a crossjunction, either with numerical simulations or with experiments. Experiments were carried out with homemade PDMS-glass microsystems involving zonal electrophoresis analysis of five derivatized amino acids. Injected amount, reproducibility, separation efficiency, and analyte discrimination were evaluated and discussed. The experimental results were successfully correlated with numerical simulations. It appeared that the dynamic injection mode is much more appropriate than the floating mode as it is faster (reduction by a factor 2 of the total analysis time here), more reproducible (RSD of peak areas equal to 1.3% (n = 4) instead of 10% (n = 4)), and leads to more efficient separation (about 20% with 3 cm separation channel length) for the same injected amount, whatever the amount, because the sample plug is less dispersed.

Analysis of carboxylic acid metabolites from the tricarboxylic acid cycle in Bacillus subtilis cell extract by capillary electrophoresis using an indirect photometric detection method

With a growing interest in metabolome analysis, there is a need for developing robust methods for analysis of intracellular metabolites profiles in real samples like e.g., bacteria cell. Due to their weak absorbance properties, tri- and dicarboxylic acids from TCA cycle (citric, isocitric, 2-oxoglutaric, succinic, fumaric, malic) as well as carboxylic acid metabolites from glycolysis pathway, urea cycle and metabolism of amino compounds (formic, pyruvic, lactic, acetic, glutamic) were analyzed by capillary electrophoresis (CE) with indirect UV detection. Using 4 m M 2,6-pyridinedicarboxylic acid as a highly UV absorbing carrier electrolyte, 0.2 m M cetyltrimethylammonium bromide, 10% ethylene glycol and 10% acetonitrile, pH 3.5, carboxylic acids metabolites were analyzed in Bacillus subtilis cell extract from two different cultures: glucose and malate. CE with an electrokinetic injection mode achieved limits of detection in the range of 13–54 ppb (1.12·10 −7–5.96·10 −7 M). The reproducibility and linearity of method was investigated with RSD for migration time less than 1.3% and acceptable correlation coefficients. The optimized CE method was used to compare metabolome content of cell extract derived from two different culture media containing either glucose or malate as a carbon source. The changes in carboxylic acid metabolites profile were observed depending from used culture medium. Carboxylic acid concentrations ranged: in cell extract from malate culture from 59 to 0.5 μ M for lactate and citrate, respectively, and in cell extract from glucose culture from 133 to 0.5 μ M for glutamate and citrate, respectively. Appropriate concentrations of carboxylic acid in the single bacterium cell were estimated at m M and sub-m M levels.

Determination of hydroxymethanesulfonic acid in environmental samples by capillary electrophoresis

Capillary electrophoresis (CE) with indirect UV detection was applied to the quantitative direct determination of hydroxymethanesulfonate ion (HMSA) present in atmospheric aerosols and other environmental samples. The proposed method used 2,6-naphthalenedicarboxylic acid (NDC)-based background electrolyte that allowed indirect detection at 214 nm. Separation was achieved by applying constant current (6.6 ± 0.6 μA which corresponds to –15 kV). According to the HMSA levels, samples were injected either by hydrodynamic (a pressure of 70 mbar, 10 s) or electrokinetic injection (–10 kV, 15 s). Method performance was determined for both injection modes. With pressure injection, the limit of detection (LOD) for HMSA was 39 μg/L and the external calibration was linear from 0.5 to 10 mg/L. With electrokinetic injection, the LOD was 2 μg/L and the internal calibration was linear from 20 to 400 μg/L. Good intra- and inter-day precision, and inter-person reproducibility and recovery were obtained for both hydrodynamic and electrokinetic injection modes. The long-term reliability of the proposed CE method was checked over a one-year period during which over 1 000 samples were analyzed.

Use of eosin as a fluorophore in capillary electrophoresis with laser detection

Eosin has been used to generate the background signal for indirect fluorimetric detection of inorganic and organic ions, simultaneously separated by capillary zone electrophoresis (CZE). This reagent provides constant fluorescence over the pH range of 5–10 and is compatible with the excitation by an argon ion laser at 488 nm with emission at 520 nm. The use of esosine as fluorophore, H 3BO 3, and Na 2B 4O 7 as electrolyte and diethylentriamine as modifier of the electroosmotic flow in CZE were optimised. The analytical potential of the studied buffer was tested on a group of 12 anions, used as model compounds. Both, hydrodynamic and electrokinetic injection mode were optimised. The detection limits determined by the last injection mode, were in the range 0.008–0.037 mg l −1. By using this method, the quantitation of the common anions in tap and mineral water has been carried out successfully.

Quantitative determination of clenbuterol, salbutamol and tulobuterol enantiomers by capillary electrophoresis.

Enantiomers of clenbuterol, salbutamol and tulobuterol were directly separated and quantitated from a spiked sample by capillary electrophoresis (CE) using sulfaited beta-cyclodextrin (SCD) as chiral selector and phosphate as running buffer. The SCD and buffer concentration, pH and field strength were the parameters studied to optimize the separation. Optimal separation was obtained using 50 mM of phosphate monobasic at pH = 2.24, 0.25% (w/w) of sulfated cyclodextrin and a field strength of 10 kV, with 20 min total time analysis. Comparison between two different injection modes (hydrodynamic and electrokinetic) was made. In the hydrodynamic mode, repeatability (expressed as relative standard deviation, RSD) was less than 1.2% for migration times for all the analyte peaks and less than 2% for peak area percentages. With respect to reproducibility, RSD was less than 3.8% for migration time and less than 3% for peak area percentages. Calibration curves were set up for two different sample concentration ranges (1 to 10 microg mL(-1) and 160-800 ng mL(-1), of each of the racemates studied). Although the electrokinetic injection mode for an aqueous sample appeared to suffer from some enantiodiscrimination, calibration curves were linear in the range between 1 and 10 ng mL(-1) with regression coefficients ranging from 0.9996 to 0.9952. As in the case of hydrodynamic injection, the method was tested with a spiked sample.

Electrokinetic injection for stacking neutral analytes in capillary and microchip electrophoresis.

An on-column mechanism for electrokinetically injecting long sample plugs with simultaneous stacking of neutral analytes in capillary electrokinetic chromatography is presented. On-column stacking methods allow for the direct injection of long sample plugs into the capillary, with narrowing of the analyte peak width to allow for an increase in the detected signal. Low-pressure injections (approximately 50 mbar) are commonly used to introduce sample plugs containing neutral analytes. We demonstrate that injection can be accomplished by applying an electric field from the sample vial directly into the capillary, with neutral analytes injected by electroosmotic flow at up to 1 order of magnitude faster than the corresponding pressure injections. Since stacking occurs simultaneously with electrokinetic injection, stacking is initiated at the capillary inlet, resulting in an increased length of capillary remaining for separation. Reproducibility obtained for peak height and peak area with electroosmotic flow injection is comparable to that obtained with the pressure injection mode, while reproducibility of analysis time is markedly improved. Electrokinetic stacking of neutral analytes utilizing electroosmotic flow is demonstrated with discontinuous (high conductivity, high mobility) as well as continuous (equal conductivity, equal mobility) sample electrolytes. Injecting neutral analytes by electroosmotic flow affords a 10-fold or greater decrease in analysis times when capillaries of 50-microm i.d. or smaller are used. This stacking method should be exportable to dynamic pH junction stacking and electrokinetic chromatography with capillary arrays. Equations describing this electrokinetic injection mode are introduced and stacking of a neutral analyte on a microchip by electrokinetic injection using a simple cross-T channel configuration is demonstrated.

Determination of metal ions in algal solution samples by capillary electrophoresis.

Trace metals determination in aqueous samples can be readily accomplished by capillary electrophoresis (CE) via indirect absorbance detection. A method for the separation of metal ions is presented and applied to the determination of seven metals in algal solution samples. 2-Hydroxyisobutyric acid background electrolyte (BGE) containing UV CAT-1 (an ultraviolet-absorbing amine) is used to perform capillary ion analysis. Acetic acid is used to adjust the pH value of BGE to 4.4. All ions can be separated in less than 15 min. All peaks are well separated and baseline resolved (i.e., no peaks overlapped). This work presents the applicability of CE to the quantitative analysis of algal solution samples and shows the adsorption process of seven metals in solution (Mn, Cd, Cr, Ni, Zn, Pb, Cu) to Chlorella vulgaris. The innovation of the application of CE in the determination of metals bound by Chlorella vulgaris is shown to be an improvement of the pH over what has been published previously. The detection limit is in the range of 13 (Mn) to 102 (Pb) ppb with electrokinetic injection mode (15 kV, 7 s). Reproducibility was 1.4% for the migration time, better than 5% for peak area for four of the metal ions (Cr, Mn, Cd, and Cu), and lower than 5% for the other three (Ni, Zn, and Pb). Calibration curves are linear for most ions in the 10(-7)-10(-5)M range (correlation coefficient r2 = 0.9933-0.9986) using electrokinetic injection mode.

Determination of metal cations by capillary electrophoresis effect of background carrier and completing agents

The determination of trace metals in aqueous samples can be readily accomplished by capillary electrophoresis (CE) via indirect absorbance detection. Methods for simultaneously determining alkali, alkaline earth and transition metal ions and Group IB, IIB and IVA metals ions were developed. Imidazole, benzylamine, ephedrine or pyridine was used as the carrier buffer and the background absorbance provider. Glycolic acid, α-hydroxy-isobutyric acid or succinic acid was used as the complexing agent. The elements determined were Li, Na, K, Cs, Mg, Ca, Sr, Ba, Cr, Mn, Fe, Co, Ni, Cu, Zn, Cd, Ag, Al and Pb. All ions could be separated in less than 15 min. In most instances reported here, more than a dozen ions could be separated in 5–10 min. All peaks were well separated and baseline resolved ( i.e., no peaks overlapped), except Ag and Al, which required a separate and additional analysis. The detection limit was in the range 0.02 (Na)−208 ppb (Cr) with the electrokinetic injection mode (10 kV, 5 s). The reproducibility was 1% for the migration time and better than 5% for the peak height for most metal ions. The calibration graphs were linear for most ions in the concentration range 10 −5−10 −3 M ( R 2 = 0.9995−0.9999) using the hydrodynamic injection mode. Concentrations lower than 10 −5 M can be determined using the electrokinetic injection mode, but the calibration graph is not linear. The methods developed here are well suited for determining metal ions in a variety of real samples.