Optimization and validation of a capillary electrophoresis methodology for inorganic anions in atmospheric aerosol samples

Optimization and validation of a method for the determination of inorganic anions, such as chloride, nitrate, sulphate, fluoride and phosphate, in atmospheric particulate matter is described. Ultrasonic extraction followed by CE, using CTAB as EOF modifier, has been established for this separation using chromate as visualization reagent for the indirect UV/vis detection. Optimization of the experimental chemical conditions (pH, surfactant concentration, chromate concentration and acetonitrile concentration) and instrumental parameters (temperature, applied voltage and injection time) is described. Linear calibrations plots are obtained for the five ions, with detection limits in the high ng/mL range. A standard reference material, SRM 1648 Urban Particulate Matter, has been used to validate the proposed method. Satisfactory reproducibility and acceptable agreement to the matter atmospheric samples has been found (recoveries ranging from 86 to 96%). Application to real particulate matter atmospheric samples, collected on high volume samplers from the air quality surveillance network of Extremadura, southwest Spain, is shown.     

Determination of anions in rainwater by capillary electrophoresis with conductivity detection

A routine method for the determination of chloride, nitrate and sulfate anions in rainwater by capillary electrophoresis was developed. The system uses an end-column non-suppressed conductivity detector. Linear calibration plots were generated from 0.050 to 20 mg/l, which is the range generally found in wet depositions. Accuracy and precision were evaluated by analyzing certified standards of simulated rainwater and environmental samples, or by comparing CE results with those obtained by IC, the reference technique for anion analysis in wet deposition. The reproducibility of the method was satisfactory except at the lower and upper limits of the analytical range. Sensitivity lay in the range of few μg/l.     

毛细管电泳法快速检测饮用水中的常见阴离子

用毛细管电泳法快速检测饮用水中常见的阴离子,并对几种电解液进行了对比试验,试验结果表明,以TTAOH(十四烷基三甲基氢氧化胺)作电渗流改进剂,pH 9.1的电解液检测效果为最佳;该方法所检离子线性相关系数均在0.999以上.     

Parameters influencing separation and detection of anions by capillary electrophoresis

Electrolyte composition is critical in optimizing separation and detection of ions by capillary electrophoresis. The parameters which must be considered when designing an electrolyte system for capillary electrophoresis include electrophoretic mobility of electrolyte constituents and analytes, detection mode, and compatibility of electrolyte constituents with one another. An electrolyte system based on pyromellitic acid is well suited for use with indirect photometric detection, and provides excellent separations of anions. The ability to modify the electrophoretic mobility of pyromellitic acid as a function of ph provides flexibility in matching electrophoretic mobilities of analytes. Additionally, the use of alkyl amines as electroosmotic flow modifiers allows the rapid separation of anions by reversing the direction of electroosmotic flow in a fused-silica capillary. The optimization of a capillary electrophoresis electrolyte for anion analysis is also discussed in terms of pH, ionic strength and applied voltage. The effect of organic solvent on separation selectivity is also discussed.     

Sensitive determination of anions in saliva using capillary electrophoresis after transient isotachophoretic preconcentration

A transient isotachophoresis-capillary electrophoresis (tITP-CE) system for the determination of minor inorganic anions in saliva is described. The complete separation and quantification of bromide, iodide, nitrate, nitrite, and thiocyanate has been achieved with only centrifugation and dilution of the saliva sample. In-line tITP preconcentration conditions, created by introduction of the plugs of 5 mM dithionic acid (leading electrolyte) and 10 mM formic acid (terminating electrolyte) before and after the sample zone, respectively, allowed the limits of direct UV absorption detection (at 200 nm) to be up to 50-fold improved as compared with CE without tITP. As a result, nitrate and thiocyanate were still detectable at 4.6 and 3.8 μg l⁻¹, respectively, in 1000 times diluted saliva. The daily variations of anionic concentrations in saliva samples taken from a smoking health volunteer were discussed based on the results of tITP-CE analysis. It was confirmed that the thiocyanate concentration in saliva noticeably increased after smoking. This is apparently the first report on simultaneous quantification of more than four anionic salivary constituents using CE.     

Cation‐enhanced capillary electrophoresis separation of atropoisomer anions

CE was used to study the separation of the atropoisomers of four phosphoric acids and two sulfonic acids and the enantiomers of two phosphoric acids. All solutes are in their anionic forms in aqueous electrolytes. The chiral additives were two hydroxypropyl cyclodextrins (CDs) and cyclofructan 6 (CF6). The CDs were able to separate four solutes and the CF6 additive could separate only one: 1,1′‐binaphthyl‐2,2′‐diyl hydrogenphosphate (BHP). Since CF6 is able to bind with cations, nitrate of alkaline metals, Ba²⁺, and Pb²⁺ were added, greatly improving the BHP separation at the expense of longer migration times. There seems to be a link between CF6–cation‐binding constants and BHP resolution factors. Cation additions were also performed with CD selectors that are less prone to form complexes with cations. Significant improvements of enantiomer or atropoisomer separations were observed also associated with longer migration times. It is speculated that the anionic solutes associate with the added cations forming larger entities better differentiated by CDs.     

Simultaneous separation of inorganic anions and cations by capillary zone electrophoresis

A new capillary electrophoretic approach for simultaneous separation of fast anions and cations is demonstrated. Indirect UV detection at 214 nm in conjunction with electromigration sampling from both ends of the capillary was developed. Two electrolyte systems based on imidazole-nitrate and copper(II)-ethylenediamine-nitrate were investigated for the simultaneous separation of chloride, sulphate, hydrocarbonate, potassium, ammonium, calcium, sodium and magnesium ions. Experimental parameters that were evaluated included a nature of UV chromophore, pH of electrolyte, a nature of complexing agent. The method permits the excellent separation of three anions and five cations in only 4 min using electrolyte system containing 2.5 mmol l⁻¹ Cu(NO₃)₂, 5 mmol l⁻¹ ethylenediamine and 1 mmol l⁻¹ fumaric acid at pH 8.5 adjusted with tetraethylammonium hydroxide.     

Ion association with alkylammonium cations for separation of anions by capillary electrophoresis

A background electrolyte (BGE) containing a 100 mM concentration of an alkylammonium cation with ethyl, propyl or butyl groups provides an excellent medium for separation of anions by capillary electrophoresis (CE). Two major effects were noted. Use of one of a series of alkylammonium cations in the BGE at a selected pH provides a simple and effective way to vary and control electroosmotic flow (EOF) over a broad range. It is believed that the alkylammonium cations are coated onto the capillary surface through a reversible dynamic equilibrium. Secondly, alkylammonium cations modify the electrophoretic migration of sample anions and the electroosmotic migration of neutral organic analytes by association interaction. This selective interaction results in improved anion separations and permits the simultaneous separation of neutral analytes. The degree of association interaction varies with the bulk and hydrophobicity of the alkylammonium cations. Incorporation of an aliphatic amine salt of moderate molecular weight in the running electrolyte provides a valuable new way to vary the migration times of sample anions and to optimize their resolution. The interactions between alkylammonium cations and sample anions or neutral organics appear to take place entirely within the liquid phase and do not require a polymeric or micellar pseudo phase.     

Method development and validation for the separation and determination of omeprazole enantiomers in pharmaceutical preparations by capillary electrophoresis

A new capillary zone electrophoresis (CZE) method for the separation of omeprazole enantiomers has been developed. Methyl-β-cyclodextrin (methyl-β-CD) was chosen as the chiral selector, and several parameters, such as cyclodextrin structure and concentration, buffer concentration, pH, and capillary temperature were investigated in order to optimize separation and run times. Analysis times, shorter than 8 min were found using a background electrolyte solution consisting of 40 mM phosphate buffer adjusted to pH 2.2, 30 mM β-cyclodextrin and 5 mM sodium disulphide, hydrodynamic injection, and 15 kV separation voltage. Detection limits were evaluated on the basis of baseline noise and were established 0.31 mg/l for the omeprazole enantiomers. The proposed method was applied to five pharmaceutical preparations with recoveries between 84 and 104% of the labeled contents.     

Ion exchange-based preconcentration for the determination of anions by capillary electrophoresis

In the present paper a capillary zone electrophoresis (CZE)-compatible preconcentration technique for anions, based on ion exchange, is described. The described preconcentration approach has found limited use until recently because of the inherent elution step that leads to contamination of the sample with eluent components. In this paper, we describe an improved anion exchange-based preconcentration technique in which contamination of the sample with the eluent constituents, which occurs during anion elution from the preconcentration column, is eliminated by on-line chemical suppression on a packed-bed suppressor column. In the present communication, the basic principles of the proposed anion enrichment system are presented. The system was optimized, resulting in a minimal additional dilution of the eluted sample plug. This was achieved by the use of a computer-controlled, sensing/switching system. The effectiveness of the developed method was later tested on the determination of some anions in a synthetic sample using CE apparatus.     

Method development and validation for the determination of various sulfur-containing anions and other anions in the corrosion process by capillary ion electrophoresis with indirect detection

A method of capillary ion electrophoresis with indirect detection is developed for the simultaneous determination of the sulfur-containing anions S2O4(2-), S2O3(2-), SO4(2-), SO3(2-), and S2- and other anions (Cl-, Br-, NO2-, NO3-, (COO)2(2-), F-, and PO4(3-)) in the corrosion process. The effects of pH, tetradecyltrimethylammonium hydroxide, chromate, 2-[n-cyclohexylamino]-ethane sulfonate, calcium gluconate, and acetonitrile on the migration and resolution of the anions and the stability of sulfur-containing anions are systematically investigated. The detection limits, repeatability, and linearity for the anions are comparatively studied at 374, 274, and 254 nm, and the results show that 374 nm is the optimal length. The simultaneous multiwavelength detection at 374, 254, 214, and 195 nm can assist in confirming the identification of UV-absorbing anions.     

Simultaneous separation of inorganic anions and cations using capillary electrophoresis with a movable contactless conductivity detector

A number of small inorganic anions and cations were separated after injection of the sample into both ends of a separation capillary. The ions were detected using a capacitively coupled contactless conductivity detector (CCCCD) which could be placed at various positions along the capillary length. Counter-directional migration of anions and cations occurs towards the detector, which is placed at an appropriate position along the capillary so that the migration order is determined by the respective effective separation capillary lengths for both anions and cations. As the CCCCD detector can be easily moved to any position along the capillary, virtually any effective separation length can be attained. Depending on the number of analytes in the sample, one can choose to obtain either electropherograms with inter-migrating zones of cations and anions or separations with distinct regions of anion and cation zones, respectively. A new term 'apparent separation selectivity' is introduced to describe the manner in which the position of the detector can be varied in order to determine the final separation.     

Routine determination of major anions in atmospheric aerosols by capillary electrophoresis

Capillary electrophoresis (CE) with indirect UV detection utilizing a pyromellitate-based electrolyte was used for the routine analysis of major anions in atmospheric aerosols collected on filters with high-volume (Hi-Vol) samplers. The long-term reliability of the CE system was checked over an 8-month period during which over 2900 samples were analyzed. In addition, approximately 1100 samples were analyzed in parallel by ion chromatography (IC). It has been shown that acceptable analytical performance can be routinely obtained. The agreement between the CE and IC results is good, generally better than 20% at concentrations larger than 1 mg l⁻¹.     

Development and validation of a fast method for determination of free glycerol in biodiesel by capillary electrophoresis

In this study, a capillary electrophoresis (CE) methodology for the determination of free glycerol in biodiesel using oxidative cleavage with periodate was optimized and validated. The amount of iodate produced in the reaction was determined by CE. The optimized electrolyte was 20 mmol L(-1) glycine and 10 mmol L(-1) trifluoroacetic acid (direct UV detection, 210 nm). The short total analysis time (less than 28 s) was obtained using the short end injection mode. The optimization of the method was carried out using Peakmaster software. The choice of the components of the run electrolyte and of the internal standard (nitrate) was made through the use of effective mobility curves. A good correlation coefficient higher than 0.9991 and low LOD 4.3 mg L(-1) was obtained. The recovery of free glycerol was 95.4-102.4%. This method was used to determine glycerol in commercial biodiesel samples.     

Development and validation of a capillary zone electrophoresis method for the quantitative determination of anthocyanins in wine

A capillary zone electrophoresis (CZE) method is proposed for the quantitative determination of anthocyanins in wine as an alternative to high-performance liquid chromatography. The CZE separation was carried out using a 46 cm (effective length)×75 μm I.D. fused-silica capillary at 10 °C and a 50 mM sodium tetraborate buffer at pH 8.4 with 15% of methanol as modifier. A voltage of 25 kV and a hydrodynamic injection of 300 mbar s were used. The electropherograms were recorded at 599 nm. It was found that SO₂ (antibacterial and antioxidant agent added to wine during its production) increased the absorbance of anthocyanins at 599 nm in a basic medium. Therefore, a concentration of 250 mg/l of SO₂ was added to the samples and the calibration solution before the analysis in order to avoid errors by this matrix effect. The analytical response was linear (R=0.998) between 10 and 700 μg/ml of malvidin-3-O-glucoside. The limit of detection and the reproducibility (as a relative standard deviation, n=11) were 1 μg/ml and 1.5%, respectively. Finally, the CZE method was validated by the analysis of synthetic wine samples (errors less than 8%) and by the comparison of the results obtained in the analysis of different monovarietal wines by CZE with those obtained by the standard HPLC method. In this comparison, a good correlation (R=0.998) with a slope of 1.005±0.044 and an intercept of −0.752±6.690 was obtained for malvidin-3-O-glucoside.     

New preconditioning strategy for the determination of inorganic anions with capillary zone electrophoresis using indirect UV detection

It is widely accepted that preconditioning procedures are indispensable in capillary electrophoresis in order to achieve reproducibility of migration times and peak areas. Several preconditioning strategies have been employed for electrophoretic determinations of inorganic anions using indirect UV detection including simple flushing with buffer or alkaline or acid pre-rinsing followed by flushing with electrolyte. We investigated the influence of various preconditioning strategies on the reproducibility of migration times and peak areas of inorganic anions. The electrolyte systems for indirect UV detection were based on pyromellitic acid and chromic acid respectively as UV absorbing probes and hexamethonium hydroxide as electroosmatic flow modifier. Preconditioning agents under investigation were electrolyte buffer, NaOH, HCl and the free acids of the UV absorbing probes. Investigations showed that reproducibility of migration times and peak areas can be significantly improved by acid pre-rinsing using the corresponding acid of the UV absorbing probes compared to preconditioning by flushing the capillary with buffer. In contrast to acid pre-rinsing using hydrochloric acid no interfering signals within the migration time window of inorganic anions under investigation can be observed. The optimized preconditioning procedure yields relative standard deviations of migration times less than 0.25% (n=10). Relative standard deviations of corrected peak areas were below 5% applying acid preconditioning using pyromellitic acid.     

Light-emitting diode-based indirect fluorescence detection for simultaneous determination of anions and cations in capillary electrophoresis

This report presents simultaneous analysis of cations and anions by capillary electrophoresis (CE) in conjunction with indirect fluorescence detection using a blue light-emitting diode (LED), based on the displacement of fluorescein with anionic EDTA-metal complexes and anions. A new focusing system combined with a plastic lens and a 40x objective was developed and used effectively to focus the diverging beam of the LED on the capillary. The optimum compositions for simultaneous analysis of metal ions and anions are the samples prepared in 5 mM borate, pH 9.2, containing 2 mM EDTA and the background electrolytes (BGEs) consisting of 5 mM borate buffer, 5 microM fluorescein, and 1 microM NaCl at pH 9.2. Using this pre-capillary complexation method, the analysis of a sample containing five metal ions and eight anions was accomplished in 8 min, with the relative standard deviation values for the migration times less than 2.0%. The peak heights against the concentrations of the metal ions and anions are linear in 10-1000 and 50-2000 microM, with correlation coefficients better than 0.998, and 0.982, respectively. The limits of detection at a signal-to-noise ratio 3 of up to 14.6 microM for formate and as low as 3.7 microM for Ni2+. The results of the analyses of pond water and a Chinese herbal soup present the advantages of this method, including simplicity, rapidity, reproducibility, and low costs.     

Cationic diazacrown ethers as selectors for the separation of inorganic anions by capillary electrophoresis

Two types of positively charged diazacrown ether derivatives involving two quaternary ammonium ions in the 12- and 18-membered rings have been investigated as selectors for the separation of inorganic anions by capillary electrophoresis. Both the cationic macrocycles largely affected the electrophoretic mobilities of analyte anions, and a group of seven inorganic anions examined in this study was completely separated in less than 2.2 min by the use of the low concentration of the additives. From the determination of ion association constants, the cationic diazacrowns were found to show greater electrostatic interaction with divalent anions than with monovalent ones.     

Instrumental validation in capillary electrophoresis and checkpoints for method validation

Capillary electrophoresis (CE) is increasingly being used in regulated and testing environments which demand validation. The design, development and production of CE instrumentation should be governed by qualifications which ensure the quality of the finished product. The vendor should therefore provide guidelines and procedures which assist the user in ensuring the adequate operation of the instrumentation and especially in designing installation qualification (IQ) and operational qualification/performance verification (OQ/PV) procedures. OQ/PV should test those functions of an instrument which directly affect the CE analysis, i.e. voltage, temperature, injection precision and detector function. In validation of CE methods care should be taken that those aspects which directly affect the precision of peak parameters are appreciated. The relationship between CE instrumentation, chemistry and validation parameters is discussed and guidelines are presented for definition of a CE method for submission to regulatory authorities.     

Field-amplified on-line sample stacking for separation and determination of cimaterol, clenbuterol and salbutamol using capillary electrophoresis

A capillary electrophoresis method, using field-amplified sample injection (FASI), was developed for separation and determination of some beta 2-agonists, such as cimaterol, clenbuterol and salbutamol. The optimum conditions for this system had been investigated in detail. The precision of the migration time, peak height and accuracy were determined in both intra-day (n = 5) and inter-day (n = 15) assays. Under the optimum conditions, the detection limits (defined as S/N = 3) of this method were found to be lower than 2.0 ng/mL for all of these three beta 2-agonists, which were much lower than that of the conventional electro-migration injection method, the enhancement factors were greatly improved to be 30-40-fold. Such lower detection limit lets this method to be suitable for determination of above-mentioned beta 2-agonists in the urine sample. The mean recoveries in urine were higher than 96.2%, 95.6% and 95.3% for cimaterol, clenbuterol and salbutamol, respectively, with relative standard deviations lower than 3.5%.