Determination of levodopa by capillary electrophoresis with chemiluminescence detection

A rapid and simple method using capillary electrophoresis (CE) with chemiluminescence (CL) detection was developed for the determination of levodopa. This method was based on enhance effect of levodopa on the CL reaction between luminol and potassium hexacyanoferrate(III) (K₃[Fe(CN)₆]) in alkaline aqueous solution. CL detection employed a lab-built reaction flow cell and a photon counter. The optimized conditions for the CL detection were 1.0 × 10⁻⁵ M luminol added to the CE running buffer and 5.0 × 10⁻⁵ M K₃[Fe(CN)₆] in 0.6 M NaOH solution introduced postcolumn. Under the optimal conditions, a linear range from 5.0 × 10⁻⁸ to 2.5 × 10⁻⁶ M (r = 9991), and a detection limit of 2.0 × 10⁻⁸ M (signal/noise = 3) for levodopa were achieved. The precision (R.S.D.) on peak area (at 5.0 × 10⁻⁷ M of levodopa, n = 11) was 4.1%. The applicability of the method for the analysis of pharmaceutical and human plasma samples was examined.     

Detection of paracetamol by capillary electrophoresis with chemiluminescence detection

Indirect detection of paracetamol was accomplished using a capillary electrophoresis-chemiluminescence (CE-CL) detection system, which was based on its inhibitory effect on a luminol-potassium hexacyanoferrate(III) (K₃[Fe(CN)₆]) CL reaction. Paracetamol migrated in the separation capillary, where it mixed with luminol included in the running buffer. The separation capillary outlet was inserted into the reaction capillary to reach the detection window. A four-way plexiglass joint held the separation capillary and the reaction capillary in place. K₃[Fe(CN)₆] solution was siphoned into a tee and flowed down to the detection window. CL was observed at the tip of the separation capillary outlet. The CL reaction of K₃[Fe(CN)₆] oxidized luminol was employed to provide the high and constant background. Since paracetamol inhibits the CL reaction, an inverted paracetamol peak can be detected, and the degree of CL suppression is proportional to the paracetamol concentration. Maximum CL signal was observed with an electrophoretic buffer of 30 mM sodium borate (pH 9.4) containing 0.5 mM luminol and an oxidizer solution of 0.8 mM K₃[Fe(CN)₆] in 100 mM NaOH solution. Under the optimal conditions, a linear range from 6.6 × 10⁻¹⁰ to 6.6 × 10⁻⁸ M (r = 0.9999), and a detection limit of 5.6 × 10⁻¹⁰ M (signal-to-noise ratio = 3) for paracetamol were achieved. The relative standard deviation (R.S.D.) of the peak area for 5.0 × 10⁻⁹ M of paracetamol (n = 11) was 2.9%. The applicability of the method for the analysis of pharmaceutical and biological samples was examined.     

Ultrasensitive determination of cobalt in single hair by capillary electrophoresis using chemiluminescence detector

By use of the high separation capability of capillary electrophoresis (CE) and ultrasensitive chemiluminescence (CL) detection, a CE-CL method was proposed for the ultrasensitive determination of trace cobalt in a single hair, which is potentially useful in construction of the fingerprint of trace elements in the hair collected from crime scenes. In this work, the CE experimental conditions, CL experimental conditions and the digestion methods for the analysis of a single hair sample for cobalt were investigated in detail. The relative limit of detection (LOD, 3σ) was 0.01 ng/mL, and the absolute LOD was 2.4 × 10^− 16 g considering the sampling volume of 24 nL. Using a dry digestion method, the analytical results for certified reference hair samples by the proposed method were in good agreement with the certified values. Finally, this method was successfully used to detect trace cobalt in a single hair from three adults. It has potential applications in forensic analysis.     

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

Chromiumexistsindifferentoxidationstatesingroundwater,industrialwastewater,seawater,andsoilofourenvironment1,2.Chromium(III)isanessentialtraceelementforhumans,requiredforthemaintenanceofnormalglucose,cholesterol,andfattyacidmetabolism.Ontheotherhand,watersolublechromium(VI),intheformCr2O72-orCrO42-,ishighlyirritatingandtoxictohumansandanimals3.Itsacutetoxiceffectsincludeanimmediatecardiovascularshockandlatereffectsonkidney,liver,andblood-formingorgans.Therefore,itisnecessaryforriskassessme…     

Determination of clenbuterol by capillary electrophoresis immunoassay with chemiluminescence detection

A competitive immunoassay for clenbuterol (CLB) based on capillary electrophoresis with chemiluminescence (CL) detection was established. The method was based on the competitive reaction of horseradish peroxidase (HRP)-labeled CLB (CLB-HRP) and free CLB with anti-CLB antiserum. The factors affecting the electrophoresis and CL detection were systematically investigated with HRP as a model sample. Under the optimal conditions, the tracer CLB-HRP and the immunoassay complex were separated, and the linear range and the detection limit (S/N = 3) for CLB were 5.0-40 nmol l⁻¹ and 1.2 nmol l⁻¹, respectively. The proposed method has been applied satisfactorily in the analysis of urine sample.     

Running buffers for determination of chromium(VI)/(III), cobalt(II) and zinc(II) in complex matrices by capillary electrophoresis with contactless conductivity detection

Complex matrices and rather high acidity in environmental samples are often the impelling challenges for the used running buffers of capillary electrophoresis. Twelve binary acid-base buffers were evaluated for separation of Cr(VI)/Cr(III), Co²⁺ and Zn²⁺ in a sample containing various salts by capillary electrophoresis with contactless conductivity detector. The malic acid (MA) systems including MA-His (histidine), MA-Arg (arginine) and MA-Tris (tris(hydroxymethyl)aminomethane) were selected as the candidates with powerful separation efficiency and good response sensitivity. In the MA-Tris buffer, optimization were further carried out in terms of the pH value and the concentration of MA, and the optimal conditions were obtained as 6 mM MA-Tris and 2 mM 18-crown-6 at pH 3.5. Furthermore, a real application was demonstrated by analyzing the plating rinse water (pH 0.8), in which the Ca²⁺, Na⁺, Cr(VI)/Cr(III), Co²⁺ and Zn²⁺ were all detected by adjusting at pH 3.5 with 5% (v/v) diluent ammonia. Both the cations, e.g., K⁺, Ca²⁺, Na⁺, Mg²⁺, and the common high concentration anions in the sample, e.g., Cl⁻, SO₄²⁻ and NO₃⁻ did not cause any disturbance to the concerned analytes.     

Speciation of chromium (III) and chromium (VI) by capillary electrophoresis with contactless conductometric detection and dual opposite end injection

A sensitive, rapid and inexpensive capillary electrophoretic method for the determination of Cr(III) and Cr(VI) species is presented. The method is based on the dual opposite end injection principle and contactless conductometric detection. The sample containing cationic and anionic species is injected into the opposite ends of the separation capillary and after the high voltage is applied, the analytes migrate towards the capillary center, where the cell of a contactless conductivity detector is placed. The method does not require any sample pretreatment, except dilution with deionized water. The separation of Cr(III), Cr(VI) and other common inorganic anions and cations is achieved in less than 4 min. The parameters of the separation electrolyte solution, such as pH and concentration of L-histidine, were optimized. Best results were achieved with electrolyte solution consisting of 4.5 mM L-histidine, adjusted to pH 3.40 with acetic acid. The detection limits achieved for Cr(III) and Cr(VI) were 10 and 39 microg.L(-1), respectively. The repeatability of migration times and peak areas was better than 0.3% and 2.8%, respectively. The developed method was applied to the analyses of rinse water samples from the galvanic industry. The results for the determination of Cr(III) and Cr(VI) were in good agreement with the results obtained by certified differential spectrophotometric method using diphenylcarbazide (CN 83 0520-40) and with the results for the total chromium concentrations determined by electrothermal atomic absorbance spectrometry (ET-AAS) and inductively coupled plasma-mass spectrometry (ICP-MS).     

Quantum dot-enhanced chemiluminescence detection for simultaneous determination of dopamine and epinephrine by capillary electrophoresis

A sensitive method based on quantum dot (QD)-enhanced capillary electrophoresis-chemiluminescence (CE-CL) detection was developed for simultaneous determination of dopamine (DA) and epinephrine (E). In this work, CdTe QD was added into the running buffer of CE to catalyze the post-column CL reaction between luminol and hydrogen peroxide, achieving higher CL emission. Negative peaks were produced due to the inhibitory effects on CL emission from DA and E eluted from the electrophoretic capillary. The decrease in CL intensity was proportional to the concentration of DA and E in the range of 8.0 × 10⁻⁸-5.0 × 10⁻⁶ M and 4.0 × 10⁻⁸-5.0 × 10⁻⁶ M, respectively. Detection limits for DA and E were 2.3 × 10⁻⁸ M and 9.3 × 10⁻⁹ M, respectively. Using this method, the levels of DA and E in human urine from healthy donors were determined.     

Separation and kinetic study of chromium(III) chloro complexes by capillary electrophoresis

Summary Three Cr(III) species (dichlorotetraaquachromium (III), [CrCl₂(H₂O)₄]⁺; monochloropentaaquachromium(III), [CrCl(H₂O)₅]²⁺; and hexaaquachromium(III), [Cr(H₂O)₆]³⁺) have been separated and determined by capillary electrophoresis. The first two complexes could be detected in direct mode in phosphate buffer, but because the absorption of complex [Cr(H₂O)₆]³⁺ is poor in the UV range, indirect UV detection had to be used. For indirect detection 5 mM imidazole was added to the buffer solution. The formation and decomposition of the different Cr(III) complexes were monitored in time after the preparation of solutions of CrCl₃.6H₂O. The slowest process was the decomposition of [CrCl(H₂O)₅]²⁺; 300 h after preparation of a solution of CrCl₃.6H₂O of pH 1 the solution contained only [Cr(H₂O)₆]³⁺. The effects of pH and the content of some matrix ions on the rates of conversion of the complexes were studied. The kinetic characteristics of this complex system could be investigated adequately by means of capillary electrophoresis. Presented at Balaton Symposium on High-Performance Separation Methods, Siófok, Hungary, September 1–3, 1999     

The speciation of rhodium(III) in hydrochloric acid media by capillary zone electrophoresis

A fast and effective method to study the aquation of rhodium(III) chlorocomplex in hydrochloric solutions using capillary zone electrophoresis (CZE) is developed. At least five species, some of which seem to be oligomeric, are formed in solution during the aquation process at pH>1. The fast hydration of RhCl₆³⁻ makes this species impossible to detect. The first species detected in the optimised conditions is RhCl₅(H₂O)²⁻ although RhCl₄(H₂O)₂⁻ is the main species during the first stage of the aquation process. When equilibrium is reached either RhCl₃(H₂O)₃ or a cationic complex, RhCl₂(H₂O)₄⁺, is formed as the main species. Matrix Assisted Laser Desorption Ionisation Time of Flight Mass Spectrometry (MALDI-TOF MS) is used as a novel technique to elucidate the structure of the rhodium aqua/chloro complexes formed in solution. Results obtained by CZE are confirmed by spectrophotometry.     

A sensitive and rapid immunoassay for quantification of CA125 in human sera by capillary electrophoresis with enhanced chemiluminescence detection

In this paper we have presented a sensitive and rapid immunoassay (IA) method by capillary electrophoresis with an enhanced chemiluminescence detection system (CE-CL) based on the catalytic effects of horseradish peroxidase (HRP) on the luminol-hydrogen peroxide reaction. The conditions for the CL reaction and electrophoresis were systematically investigated using HRP as a model sample. The linear range from 2.5 x 10(-11) to 1.0 x 10(-9) mol/L (R = 0.999), and the detection limit of 1.0 x 10(-12) mol/L (signal-to-noise ratio = 3) for HRP were achieved using para-iodophenol as CL enhancer. The relative standard deviations of the migration time and peak area for 5.0 x 10(-10) mol/L HRP (n = 7) were 0.26 and 4.8%, respectively, using a CE system with a home-built CL detector. Under the optimal condition, the HRP-labeled CA125 antibody (Ab) and the Ab-antigen complex were well separated within 4 min by CE using a high-pH buffer (pH 10.20). The assay was successfully used for quantification of CA125 in human sera from health controls and patients associated with ovarian cancer, and the recoveries of the standard addition experiments were 93-109%. Our primary results demonstrated that IA based on CE-CL detection is a powerful tool for clinical diagnosis combined with these commercial IA kits.     

Speciation of Cr(III) and Cr(VI) in waters using immobilized moss and determination by ICP-MS and FAAS

The possibility of using moss (Funaria hygrometrica), immobilized in a polysilicate matrix as substrate for speciation of Cr(III) and Cr(VI) in various water samples has been investigated. Experiments were performed to optimize conditions such as pH, amount of sorbent and flow rate, to achieve the quantitative separation of Cr(III) and Cr(VI). During all the steps of the separation process, Cr(III) was selectively sorbed on the column of immobilized moss in the pH range of 4-8 while, Cr(VI) was found to remain in solution. The retained Cr(III) was subsequently eluted with 10 ml of 2 mol l⁻¹ HNO₃. A pre-concentration factor of about 20 was achieved for Cr(III) when, 200 ml of water was passed. The immobilized moss was packed in a home made mini-column and incorporated in flow injection system for obtaining calibration plots for both Cr(III) and Cr(VI) at low ppb levels that were compared with the plots obtained without column. After separation, the chromium (Cr) species were determined by inductively coupled plasma mass spectrometry (ICP-MS) and flame atomic absorption spectrometry (FAAS). The sorption capacity of the immobilized moss was found to be ∼11.5 mg g⁻¹ for Cr(III). The effect of various interfering ions has also been studied. The proposed method was applied successfully for the determination of Cr(III) and Cr(VI) in spiked and real wastewater samples and recoveries were found to be >95%.     

癌胚抗原毛细管电泳-化学发光均相免疫分析

设计了一种测定人血清中癌胚抗原的毛细管电泳-化学发光检测均相免疫分析新方法。采用四苯硼酸钠增强luminol-H2O2-HRP体系化学发光的原理,化学发光检测经毛细管电泳分离的,用辣根过氧化物酶(HRP)标记的癌胚抗体及免疫复合物。测定癌胚抗原的线性范围2.0~80.0μg/L(R=0.9921),检出限为0.1μg/L(绝对检出限为0.75 fg)。     

Speciated isotope dilution analysis of Cr(III) and Cr(VI) in water by ICP-DRC-MS

An isotope dilution method has been developed for the speciation analysis of chromium in natural waters which accounts for species interconversions without the requirement of a separation instrument connected to the mass spectrometer. The method involves (i) in-situ spiking of the sample with isotopically enriched chromium species; (ii) separation of chromium species by precipitation with iron hydroxide; (iii) careful measurement of isotope ratios using an inductively coupled plasma mass spectrometer (ICP-MS) with a dynamic reaction cell (DRC) to remove isobaric polyatomic interferences. The method detection limits are 0.4 μg L⁻¹ for Cr(III) and 0.04 μg L⁻¹ for Cr(VI). The method is demonstrated for the speciation of Cr(III) and Cr(VI) in local nullah and synthetically spiked water samples. The percentage of conversion from Cr(III) to Cr(VI) increased from 5.9% to 9.3% with increase of the concentration of Cr(VI) and Cr(III) from 1 to 100 μg L⁻¹, while the reverse conversion from Cr(VI) to Cr(III) was observed within a range between 0.9% and 1.9%. The equilibrium constant for the conversion was found to be independent of the initial concentrations of Cr(III) and Cr(VI) and in the range of 1.0 (at pH 3) to 1.8 (at pH 10). The precision of the method is better than that of the DPC method for Cr(VI) analysis, with the added bonuses of freedom from interferences and simultaneous Cr(III) determination.     

Speciation of oxidation states of elements by capillary electrophoresis

Progress made in the last five years in the application of capillary electrophoresis methods to chemical speciation of elements is reported on the basis of over 100 literature references. The main trends observed include development of new on‐ and off‐capillary derivatization methods, application of new detection methods, and especially coupling of CE separation systems to powerful atomic spectroscopy and mass spectrometry instruments with various ionization techniques, providing either a sensitive element‐specific detection method or a third dimension for high performance separation. Besides numerous CZE and MEKC capillary electrophoresis methods only very few examples of CE speciation with capillary electrochromatography can be found. Concerning the chemical forms of elements determined, the new procedures developed are mostly focused on redox speciation of various oxidation states of elements, metal‐bound high molecular compounds, and organometallic species.     

癌胚抗原毛细管电泳-化学发光均相免疫分析

建立了一种测定人血清中癌胚抗原的毛细管电泳-化学发光检测的均相免疫分析新方法.采用四苯硼钠增强luminol-H2 O2-HRP体系化学发光的原理,化学发光检测经毛细管电泳分离的,用辣根过氧化物酶(HRP)标记的癌胚抗体及免疫复合物.测定癌胚抗原的线性范围2.0～80.0 μg/L(R=0.9921),检出限为0.1 μg/L(绝对检出限为0.75 fg).     

Simultaneous determination of chromium(VI) and chromium(III) at trace levels by adsorptive stripping voltammetry

A new methodology was proposed for the speciation of chromium by differential pulse adsorptive stripping voltammetry (DPAdSV) using pyrocatechol violet (PCV) and N-(2-hydroxyethyl)ethylenediamine-N,N′,N′-triacetic acid (HEDTA) as complexing agents. In this procedure, a partial least squares (PLS) regression was used for the resolution of the strongly overlapping voltammetric signals from mixtures of Cr(III) and Cr(VI) in the presence of PCV and HEDTA. The relative error in absolute value was <6% when concentrations of several mixtures were calculated. The analysis of the possible effect of the presence of foreign ions in the solution was performed. The procedure was successfully applied to the speciation of chromium in different samples of natural water.     

Analysis of antioxidants using a capillary electrophoresis with chemiluminescence detection system

We developed a capillary electrophoresis with chemiluminescence detection system using 2-methyl-6-p-methoxyphenylethynylimidazopyrazinone as a chemiluminescence reagent for determination of antioxidants of superoxide anions. 2-Methyl-6-p-methoxyphenylethynylimidazopyrazinone reacted with superoxide anions generated through the reaction of hypoxanthine and xanthine oxidase, and then emitted chemiluminescence. Suppression of the chemiluminescence in the presence of antioxidants for superoxide anions was introduced as a detection principle for antioxidants into the capillary electrophoresis with chemiluminescence detection system. After optimizing the analytical conditions, various antioxidants, such as superoxide dismutase, nitroblue tetrazolium, ascorbic acid, and catechin, were subjected to the present system. They gave negative peaks due to the quenching effect; the detection limits of superoxide dismutase, nitroblue tetrazolium, ascorbic acid, and catechin were 1, 100, 100, and 10 μM, respectively (S/N = 2). A model sample consisting of superoxide dismutase and nitroblue tetrazolium was satisfactorily separated and detected within ca. 10 min. We also applied the present system to analysis of catechin in green tea as a real sample.