Progress in amperometric detection for capillary electrophoresis

As a high efficiency separation technique, capillary electrophoresis(CE) has been widely used in various fields of analytical science. Amperometry is one of the most sensitive electrochemical detection methods in CE. The capillary/electrode decoupling mechanism, applications,of new electrode systems in CE, detection cell technique are discussed in detail. Amperometric detection is compatible with microfabricated CE chips and will make the concept of lab-on-a-chip become a reality. Because of these progresses, amperometry is becoming a widely acceptable detection method,for more chemical and biological analytes.

Determination of sulpiride by capillary electrophoresis with end-column electrogenerated chemiluminescence detection

Background: Capillary electrophoresis (CE) with tris(2,2'-bipyridyl)ruthenium(II) [Ru(bpy)(3)(2+)]-electro-generated chemiluminescence (ECL) detection is a promising method for clinical analysis. In this study, a method combining CE with Ru(bpy)(3)(2+) ECL (CE-ECL) detection that can be applied to amine-containing clinical species was developed, and the performance of CE-ECL as a quantitative method for determination of sulpiride in human plasma or urine was evaluated. Methods: Sulpiride was separated by capillary zone electrophoresis in uncoated fused-silica capillaries [510 cm x 25 mum (i.d.)] filled with phosphate buffer (pH 8.0 and a driving voltage of +15 kV, with end-column Ru(bpy)(3)(2+) ECL detection. A platinum disc electrode was used as working electrode. Sulpiride in human plasma or urine samples (100 muL) was extracted by a double-step liquid-liquid extraction procedure, dried under nitrogen at 35 degreesC in a water bath, and reconstituted with 100 muL of filtered water. The extraction solvent was ethyl acetate-dichloromethane (5:1 by volume). Results: Under optimum conditions (pH 8.0 phosphate buffer, injection for 6 s at 10 kV, and +1.2 V as detection potential), separation of sulpiride was accomplished within 4 min. The calibration curve was linear over a concentration range of 0.05-25.0 mumol/L, and the limit of detection was 2.9 x 10(-8) mol/L for sulpiride. Intra- and interday CVs for ECL intensities were <6%. Extraction recoveries of sulpiride were 95.6-101% with CVs of 2.9-6.0%. The method was,clinically validated for patient plasma and urine samples. Conclusions: CE combined with Ru(bpy)(3)(2+) ECL is reproducible, precise, selective, and enables the analysis of sulpiride in human plasma and urine. It thus is of value for rapid and efficient analysis of amine-containing analytes of clinical interest.

Determination of three beta-blockers by capillary electrophoresis with end-column electrochemical detection

Three beta -blockers (propranolol, timolol, acebutolol) were separated by capillary electrophoresis (CE) and detected by end-column electrochemical detection (EC). In the present work, a carbon fiber (33 mum) electrode was used as the working electrode. The effect of the buffer concentration, buffer pH, detection potential and separation voltage on the separation of analytes and behavior of electrochemical detection was systematically investigated. The optimum conditions determined were as following: 40 cm length, 25 mum i.d. capillary; 17.5 kV separation voltage; 2 s injection at 15 kV; 70mM phosphate buffer, pH 3.5; detection potential + 1.2V (vs. Ag/AgCl). Under these conditions, the linear ranges of beta -blockers were over three orders of magnititude and the low detection limit of 10(-8)M was obtained. This method was also applied to detect the simulated urine sample.

Simultaneous determination of 2-aminothiazole, 2-aminobenzothiazole and 2-mercaptobenzothiazole by capillary electrophoresis with end-column amperometric detection

Capillary electrophoresis with amperometric detection is evaluated for the simultaneous determination of 2-aminothiazole (A), 2-amino-benzothiazole (AB), 2-mercaptobenzothiazole (AM). The cyclic voltammogram, hydrodynamic voltammogram, effect of pH, concentration of buffer and separation voltage on the separation and the detection were studied. The conditions were optimized as follows: 50 mM phosphate buffet; pH 6.0, 2s at 17.5 kV sample injection, separation at 17.5 kV, 1.2 V as detection potential. The method provided low detection limit as 0.5 mu M, 0.05 mu M and 0.01 mu M, wide linear range 2-200 mu M, 10-200 mu M and 0.025-100 mu M for A, AB, and AM, respectively. The variations in peak current and migration time for 15 consecutive injections of a standard containing 5 mu M each compound were 3.7, 2.1, and 3.9%, and 1.2, 0.8, and 1.2%, for A, AB and AM, respectively. This method was employed to analyze river water.

Determination of propranolol by capillary electrophoresis with end-column amperometric detection

A capillary electrophoresis-amperometric detection system was developed for the determination of propranolol (PRO) at a 33 mu m carbon fiber microdisk electrode (CFE). The cyclic voltammogram, the hydrodynamic voltammograms and the effect of pH were studied. Under the optimum conditions: separation Voltage 15 kV; injection 3 s at 15 kV; 10 mM pH 7.5 phosphate buffer, 1.15 V (vs. Ag/AgCl) detection potential, the detection limit (LOD) for PRO was 0.05 mu M (S/N = 3). The response for PRO was linear over two orders of magnitude with a linear correlation coefficient of 0.994. The feasibility of this method was demonstrated by the detection of PRO in urine sample.

Determination of barbituric acid and 2-thiobarbituric acid with end-column electrochemical detection by capillary electrophoresis

Capillary electrophoresis (CE) with end-column electrochemical detection (EC) of barbituric acid (BA) and 2-thiobarbituric acid (TA) has been described. Under optimum condition, BA and TA were separated satisfactorily, and a response of high sensitivity and stability was obtained at a detection potential of 1.25 V versus Ag/AgCl. Optimized end-column detection provides detection limit as low as 0.5 and 0.1 mu M for BA and TA, respectively. The calibration graph was linear over three orders of magnitude. The relative standard deviations (n = 10) of peak currents and migration times obtained for both BA and TA were 3.4, 3.7, and 1.7, 1.2%, respectively. The proposed method has been applied to analyze water sample with satisfactory results. (C) 2000 Elsevier Science B.V. All rights reserved.

Separation of phenothiazines in aqueous and non-aqueous capillary electrophoresis

Four phenothiazines, promethazine, dioxypromethazine, chlorpromazine, and trifluoperazine have been separated by capillary electrophoresis using N, N, -dimethylformamide (DMF) as separation medium with UV absorbance detection. High voltage and concentrated buffer were used with small current and low electroosmosis. Good resolution and high column efficiency were obtained. Separation selectivity in DMI; was different from that in water because of the different solvation interactions. The influence of buffer composition on separation selectivities and electroosmosis were also studied.

Chiral separation of promethazine by capillary electrophoresis with end-column amperometric detection

Sensitive end-column amperometric detection has been successfully coupled to capillary electrophoresis for chiral separation of promethazine, with a carbon fiber microdisk electrode as working electrode. Baseline separation and sensitive detection were achieved under optimum conditions: 0.030 M Na2HPO4 and 0.015 M citric acid at pH = 2.50, 1.0 mM beta -CD, 10 kV separation voltage, and detection potential 1.10 V (vs Ag/AgCl). The numbers of theoretical plates were higher than 700000, and the detection limit was 5 x 10(-8) M. On-line treatment of the electrode has also been studied and discussed.

Determination of allopurinol and its active metabolite oxypurinol by capillary electrophoresis with end-column amperometric detection

A simple method was proposed for the separation of allopurinol (AP) and its active metabolite oxypurinol (OP) by capillary electrophoresis with end-column amperometric detection. A running buffer composed of 15 rum Na2HPO4/NaH2PO4 at a pH 9.55, electrokinetic injection 7 s at 5 kV, separation voltage at 15 kV and detection potential at 1.20 V were investigated to be the optimal condition for the separation. The method exhibited low detection Emit (S/N = 3) as 1 x 10(-8) mol/l for AP and OP, wide linearity range of 2 x 10(-7) to 1 x 10(-4) mol/l, 1 x 10(-7) to 1 x 10(-4) and high efficiency of 1.2 x 10(5) and 1.8 x 10(5) N/m for AP and OP, respectively. The potential application examined for the method was the determination of the spiked urine sample, which was proved to be sensitive and efficient. (C) 2001 Elsevier Science B.V. All rights reserved.

New microsystem of capillary electrophoresis with amperometric detection

We have made a cheap microsystem of capillary electrophoresis with a new method, integrating the electrodes, injection channel, separation channel, buffer reservoirs and detection cell on a polymethylmethacrylate (PMMA) chip. Using an integrated micro carbon fiber disk electrode as the working electrode in three electrodes system, 1 x 10(-4) mol/L dopamine(DA) could be detected with end-column amperometric detection. The reproducibility was good. Peak current was 6.73 nA,theoretical plate number was 71300/m and height equivalent of one theoretical plate height was 14.0 mum for 1 x 10(-4) mol/L DA. The limit of detection was 3.6 x 10(-8) mol/L and the linear range was extended from 5 x 10(-7) mol/L to 1 x 10(-4) mol/L for DA. 1 x 10(-4) mol/L catechol (CA) and 5 x 10(-5) mol/L DA were also separated completely with R-s = 10.1.

End-column amperometric detection of aesculin and aesculetin by capillary electrophoresis

Determination of aesculin (AL) and aesculetin (AT) by capillary electrophoresis with end-column amperometric detection using a 33 mu m microdisk carbon fiber electrode is described. The HDVs, the effect of pH, buffer concentration, injection voltage, injection time and separation voltage on the peak current response (i(p)) of the analytes and the number of theoretical plates (N) were studied. The method has high sensitivity and good reproducibility. Under the optimum condition - 10 mM, pH 9.00 phosphate buffer, 4 s at 9 kV injection, separation at 15 kV and +1.0 V as the detection potential - low detection limits (S/N = 3) of 0.06 and 0.3 mu M were obtained for AL and AT, respectively. The calibration curve was linear over three orders of magnitude. The relative standard deviations (n = 15) of peak current and migration time were 3.9% and 4.6%, and 0.96% and 0.75% for 15 consecutive injections of 5 mu M AL and AT, respectively. The use of this method for the separation and detection of the two compounds present in the traditional Chinese medicine and human urine samples is also reported. (C) 1999 Elsevier Science B.V. All rights reserved.

End-column electrochemical detection for aromatic amines with high performance capillary electrophoresis

Electrochemical detection of five species of aromatic amines at a carbon fiber microdisk electrode after separation by capillary electrophoresis is described. Under the optimum conditions, the detection limit for 3,4-dihydroxybenzylamine, N,N-dimethylaniline, p-phenylenediamine, p-aminophenol and aniline sulfate was 0.9, 0.03, 0.075, 1.2 and 0.15 mu M (S/N = 3), respectively. The linear response range was 5-1000, 0.1-500, 0.5-500, 5-500 and 1-200 mu M, respectively The effect of the electrode position and buffer pH on the detection was also studied. This method is very simple, sensitive and stable for the detection of these compounds.

Combination of amperometric detector & UV detector for capillary electrophoresis

A new electrochemical cell assembly with the combination of UV and amperometric detector (AD) based on their complementarity was described. A Nafion tubing junction was used to decouple the high voltage from the separation capillary in the rear of on-column UV detector. In this mode, the electroactive and inert compounds could be detected by UV and AD at the same time. Aromatic amines were determined with the UV and the end-column AD detection to evaluate the performances of the cell assembly. Such an improved electrochemical detector could match the capillary with different diameters. By simple adjustment of the screws, the positioning of the working electrode and the detection capillary was easily gained without microscope. It is also very easy to assemble and disassemble the working electrode when needed. (C) 1999 Elsevier Science B.V. All rights reserved.

Applications of microelectrodes in capillary electrophoresis electrochemical detection

The applications of the microelectrodes for capillary electrophoresis/electrochemical detection are reviewed. The attractive advantages of the microelectrode provide a wide scope for the developments in capillary electrophoresis with electrochemical detection.

Separation of phenothiazines using aqueous and nonaqueous capillary electrophoresis

Separations of phenothiazines, promethazine(PZ), dioxypromethazine (OZ), chlorpromazine(CZ), trifluoperazine(TfZ) and thioridazine(TZ) by capillary electrophoresis in water and FA media were carried out and compared. Thus different selectivity and resolution were observed as media varying from water to FA. Migration order was PZ, OZ, CZ and TZ in water but (TZ+PZ), CZ and OZ in FA, when the same buffer, 25 mmol/L Tris and 25 mmol/L citric acid, was used. It also has been observed that pH has great effect on selectivity both in water and FA and a possible explanation was given. Separation efficiency was higher in FA media than in water because of the permission of high voltage applied. For all separations in FA 30 kV was applied, and when 25 mmol/L Tris was used as buffer, current was only 20 mu A and complete separation of TZ, CZ, PZ and OZ was achieved with effencicy higher than 3.5 x 10(5) theoretical plates for all analytes. The high performance of capillary electrophoresis in FA suggests that FA is an excellent media separation.