Determination of dyes in foodstuffs by capillary zone electrophoresis

A rapid method based on capillary zone electrophoresis coupled with photodiode-array detection has been developed to determine the dyes Tartrazine E-102, Sunset Yellow FCF E110, Amaranth E-123, New Coccine E-124, Patent Blue V calcium salt E-131 and Allura Red AC E-129 in foodstuffs. Separation was done by using a Bare CElect-FS75 CE column, using a 10 mM phosphate buffer at pH 11.0. Hydrodynamic injections at 0.5 p.s.i. for 4 s (21 nl of sample) and 20 kV separation voltage were used. The quantitation limits for the six dyes ranged from 3 to 6 microg/ml. A linear relationship between 3 to 95 microg/ml, with correlation coefficient better than 0.995 was obtained. This method has been applied to the determination of the studied dyes in beverages, jellies and syrups.     

Determination of sulphonate dyes in water by ion-interaction high-performance liquid chromatography

An ion-interaction high-performance liquid chromatography method for quick separation and determination of the sulphonated dyeAcid Yellow 1, and the sulphonated azo dyes Acid Orange 7, Acid Orange 12, Acid Orange 52, Acid Red 2, Acid Red 26, Acid Red 27 and Acid Red 88 has been developed. An RP-ODS stationary phase is used, and the mobile phase contains an acetonitrile-phosphate buffer (27:73, v/v) mixture at pH 6.7, containing 2.4 mM butylamine as ion-interaction reagent. Good separations were obtained using isocratic elution and spectrophotometric detection at 460 nm. The detection limits for the eight dyes ranged from 7 to 28 microg/l for an injection volume of 100 microl. Spiked tap water samples (100 ml), containing different concentration levels (0.3-1.2 microg/l) of the dyes were analyzed after acidification (pH 3) and preconcentration in disposable solid-phase extraction C18 cartridges.     

Development and optimization of a method for the analysis of low-molecular-mass organic acids in plants by capillary electrophoresis with indirect UV detection

Capillary zone electrophoresis was developed to analyze low-molecular-mass organic acids including oxalic, tartaric, formic, malic, citric, succinic, glutaric, acetic and lactic acid. The influences of some crucial parameters such as buffer concentration, pH value, surfactant concentration and separation temperature, on electrophoretic separation were investigated. Under the conditions of 15 mM phthalate containing 0.6 mM tetradecyltrimethylammonium bromide as the run buffer (pH 5.6); separation voltage -15 kV (-263.2 V/cm) and temperature 25 degrees C, a satisfactory separation of nine organic acids was accomplished within 7 min. The detection limits (S/N=3) ranged from 0.008 to 0.08 microg/ml and the quantification limits ranged from 0.01 to 0.1 microg/ml for electrokinetic injection. The method was successfully applied to analyze organic acids in different parts of Var splendens (an edible vegetable in China). The recoveries of organic acids in real samples ranged from 88 to 121%.     

Analysis of glyphosate and aminomethylphosphonic acid by capillary electrophoresis with electrochemiluminescence detection

A capillary electrophoresis (CE) method coupled with electrochemiluminescence (ECL) detection for the analysis of glyphosate (GLY) and its major metabolite aminomethylphosphonic acid (AMPA) is presented. Complete separation of GLY and AMPA was achieved in 8 min using a background electrolyte of 20 mM sodium phosphate (pH 9.0) and a separation voltage of 21 kV. ECL detection was performed with an indium tin oxide (ITO) working electrode bias at 1.6 V (vs. a Pt-wire reference) in a 30 0mM sodium phosphate buffer (pH 8.0) containing 3.5mM Ru(bpy)3 2+ (where bpy=2.2'-bipyridyl). Linear correlation (r>or=0.997) between ECL intensity and analyte concentration was obtained in the ranges 0.169-16.9 and 5.55-111 microg ml(-1) for GLY and AMPA, respectively. The limits of detection (LODs) for GLY and AMPA in water were 0.06 microg ml(-1) and 4.04 microg ml(-1), respectively. The developed method was applied to the analysis of GLY in soybeans. The LOD of GLY in soybean was 0.6 microg g(-1). Total analysis time including sample pretreatment was less than 1h.     

Determination of textile dyes by means of non-aqueous capillary electrophoresis with electrochemical detection

Eight textile dye compounds including five cationic dyes, namely, basic blue 41, basic blue 9, basic green 4, basic violet 16 and basic violet 3, and three anionic dyes, acid green 25, acid red 1 and acid blue 324, were separated and detected by non-aqueous capillary electrophoresis (NACE) with electrochemical detection. Simultaneous separations of acid and basic dyes were performed using an acetonitrile-based buffer. Particular attention was paid to the determination of basic textile dyes. The optimized electrophoresis buffer for the separation of basic dyes was a solvent mixture of acetonitrile/methanol (75:25, v/v) containing 1 M acetic acid and 10 mM sodium acetate. The limits of detection for the basic dyes were in the range of 0.1–0.7 μg mL⁻¹. An appropriate solid-phase extraction procedure was developed for the pre-treatment of aqueous samples with different matrices. This analytical approach was successfully applied to various water samples including river and lake water which were spiked with textile dyes.     

Determination of gamma-hydroxybutyric acid in clinical samples using capillary electrophoresis with contactless conductivity detection

A method for the determination of gamma-hydroxybutyric acid (GHB) in urine and serum samples with capillary electrophoresis using capacitively coupled contactless conductivity detection (CE-C(4)D) was developed. The optimized separation buffer consisting of 20 mM of arginine, 10 mM of maleic acid and 30 microM of cetyltrimethylammonium bromide (CTAB) contained 5 mM vancomycin to facilitate the separation of gamma-hydroxybutyric acid from beta-hydroxybutyric acid (BHB), which is also present in clinical samples. The detection limits in the clinical samples were found to be about 2 microg/ml, which is well below the required sensitivity for the recognition of overdosage and adequate for the determination of endogenous concentrations in urine. The determination of GHB in both types of samples was carried out directly after a fourfold dilution without requiring any derivatization or extraction procedures.     

Determination of water-soluble UV-filters in sunscreen sprays by liquid chromatography

Liquid chromatography was used for the determination of the three most used water-soluble UV filters, benzophenone-4 (BZ4), terephthalylidene dicamphor sulfonic acid (TDS), and phenylbenzimidazole sulphonic acid (PBS), in aqueous sunscreen sprays. A C18 stationary phase and an isocratic mobile phase of EtOH-20 mM sodium acetate buffer of pH 4.6 (30:70, v/v) were used at a flow-rate of 0.5 ml min(-1). Mobile phase was also used as solvent for samples and standards. UV detection was at 313 nm. The analytical run took 5.5 min. The limits of detection were 0.5, 0.9 and 2 microg ml(-1) for BZ4, TDS and PBS, respectively. The proposed method does not involve highly toxicsolvents.     

Determination of poly-beta-hydroxybutyric acid in Bacillus thuringiensis by capillary zone electrophoresis with indirect ultraviolet absorbance detection

A new capillary electrophoresis method for determining poly-beta-hydroxybutyric acid (PHB) in Bacillus thuringiensis was established. Poly-beta-hydroxybutyric acid in samples was hydrolyzed by sulphuric acid and neutralized by Ba(OH)2. The content of produced beta-hydroxybutyrate was then determined by capillary zone electrophoresis (CZE) with indirect UV detection at 254 nm. With 5 mM p-hydroxybenzoate and 0.5 mM tetradecyltrimethylammonium bromide (TTAB) at pH 8.0 as carrier electrolyte, beta-hydroxybutyrate can be determined within 6 min. Standard regression equation was made by beta-hydroxybutyrate, and the linear range was 2-1000 microg/ml. The relative standard deviations (RSDs) for migration time and peak area are both less than 1.0%. The detection limit for beta-hydroxybutyrate was 0.2 microg/ml, which is two to three orders of magnitude lower than that of the gas chromatography (GC) method. The capillary electrophoresis method was successfully applied to determine poly-beta-hydroxybutyric acid in fermentation broth and single colony. The added standard recovery was 96%.     

Analysis of ethambutol and methoxyphenamine by capillary electrophoresis with electrochemiluminescence detection

A capillary electrophoresis (CE) coupled with electrochemiluminescence (ECL) detection method for the analysis of ethambutol (EB) and methoxyphenamine (MP) has been investigated. Complete separation of EB and MP was achieved in 8 min using a background electrolyte of 20 mM sodium phosphate at pH 10.0 and a separation voltage of 9 kV. ECL detection was performed with an indium/tin oxide (ITO) working electrode biased at 1.4 V (versus a Pt wire reference) in a 200 mM sodium phosphate buffer (pH 8.0) containing 3.5 mM Ru(bpy)3(2+) (where bpy = 2,2'-bipyridyl). Linear correlation (r > or = 0.993) between ECL intensity and drug concentration was obtained in the range 2-50 ng/ml. The limits of detection (LODs) for EB and MP in water were 1.0 and 0.9 ng/ml, respectively. The relative standard deviation values on peak size (10 ng/ml level) and migration time for the two drugs were in the ranges 5-8 and 0.2-0.7% (n = 7), respectively. Applicability of the CE-ECL method to the analysis of human plasma spiked with EB and MP was examined. The LODs for EB and MP in plasma were 0.4 and 0.3 microg/ml, respectively.     

Determination of valproic acid (2-propylpentanoic acid) in human plasma by capillary electrophoresis with indirect UV detection

A rapid capillary zone electrophoresis method with indirect UV detection was developed and validated for the determination of valproic acid (VPA) in human plasma. The analyses were carried out under optimized conditions, using a buffer system composed of 15 mM benzoate and 0.5 mM cetyltrimethylammonium bromide at pH 6.0, and 25% v/v methanol; 2-hydroxybutyric acid was selected as the internal standard (IS). The capillary electrophoresis (CE) separation was carried out at a negative potential of 30 kV and the indirect UV detection was operated at 210 +/- 20 nm for all assays. The influence of buffer pH, ionic strength, concentration of electroosmotic flow (EOF) modifier and organic modifier on indirect signal response and migration behavior of the organic acid was investigated. Isolation of VPA from plasma was accomplished by a carefully implemented procedure using methanol as the precipitant agent. Using a high ratio of methanol to plasma for deproteinization (4:1), good absolute recovery of the analyte and satisfactory selectivity was obtained. The calibration line for VPA was linear over the 1-100 microg/mL concentration range. Sensitivity was high; in fact, the limit of detection (LOD) of VPA was 150 ng/mL and 450 ng/mL the limit of quantitation (LOQ). The results obtained analyzing real plasma samples from schizophrenic patients under polytherapy with VPA as well as antipsychotic drugs were satisfactory in terms of precision, accuracy and sensitivity.     

Identification and quantification of natural isoxazolinone compounds by capillary zone electrophoresis.

A capillary zone electrophoresis (CZE) method that is specific, simple, rapid and also cheap was developed to analyse some natural UV-absorbing isoxazolinone compounds with toxic potential present in legume seedlings. The six most common natural isoxazolinone compounds were separated within 10 min with 25 mM potassium phosphate (pH 7.5) containing 8% 1-propanol as running buffer. A 60 cm coated fused-silica capillary (52.6 cm effective length x 75 microm I.D.), with an electric field of 375 V/cm at 30 degrees C was used. The limit of detection ranged from 0.01 mM (3.0 microg/ml) to 0.03 mM (7.7 microg/ml). Linearity between peak areas and concentrations ranging from 0.05 mM to 1.75 mM were determined for each isoxazolinone. The correlation coefficient was 0.9954 or greater. Both relative migration time and peak area were reproducible. The RSD of relative migration time is between 0.44 and 1.94% and RSD of peak area is between 1.26 and 6.86%. The concentrations of isoxazolinones in Lathyrus odoratus and L. sativus seedlings obtained by CZE were in agreement with the previous results from HPLC.     

1,3-dialkylimidazolium-based room-temperature ionic liquids as background electrolyte and coating material in aqueous capillary electrophoresis

The 1-ethyl-3-methylimidazolium (EMIM) cation was found to have constant mobility of 4.5 x 10(-4) cm2 V(-1) s(-1) over the pH range of 3 to 11. The electroosmotic flow of bare silica capillary was reversed by the covalently bonded room-temperature ionic liquid (RTIL) coating. With run buffer of 5 mM EMIM (pH 8.5), NH4+ in human urine was separated from the K+ matrix and was detected to be 0.37 +/- 0.012%. K+, Na+, Li+, Ca2+, Mg2+ and Ba2+ were baseline separated in RTIL-coated capillary with run buffer of 10 mM EMIMOH-acetic acid at pH 5, and the concentration of the above ions in a red wine were detected to be 907, 27.9, 0, 71.0, 83.4 and 31.1 microg/ml, respectively. The RTIL-coated capillary showed stable electroosmotic flow for at least 80 h in the run buffer.     

Study of biodegradation products from azo dyes in fungal degradation by capillary electrophoresis/electrospray mass spectrometry

Biodegradation products from four model sulfonated azo dyes Orange II, Acid Orange 8, Food Yellow 3, and 4-[(4-hydroxyphenyl)azo]-benzenesulfonic acid, sodium salt (4HABA), during fungal degradation were determined by capillary electrophoresis coupled with ion trap mass spectrometry (CE-MS) with electrospray ionization and a coaxial sheath flow interface. The development and optimization of this analytical method including the sheath liquid composition and flow rate, nebulizing gas flow rate, carrier electrolyte, and MS voltage are described herein. Detection of unknown biodegradation products was carried out under negative ion mode with base peak electrophorogram (BPE) or extractive ion electrophorogram (EIE) monitoring. A volatile ammonium acetate buffer (10 mM) without organic modifier and a shealth liquid made from 2-propanol and water (80:20, v/v) were suited for the separation and ESI interface. The sulfonated ion was the base peak for model azo dyes and their metabolites containing sulfonic group. Results showed that the tested azo dyes were degraded quickly in the culture of white rot fungus, Pleurotus ostreatus in 3 days with the major biodegradation products being 4-hydroxy-benzenesulfonic acid, 3-methyl-4-hydroxy-benzenesulfonic acid, benzenesulfonic acid, 1,2-naphthoquinone-6-sulfonic acid and 3-methyl-benzenesulfonic acid.     

Simultaneous determination of iron(II) and iron(III) by micellar electrokinetic chromatography using an off-line selective complexing reaction.

A simultaneous determination with UV detection/capillary electrophoresis for Fe(II) and Fe(III) was achieved using a sulfonated bathophenantholine and desfferioxamine B. When the electrophoretic buffer was 60 mmol l(-1) SDS, 10 mmol l(-1) acetic acid (pH 4.0) and 10 mmol l(-1) ascorbic acid and at -10 kV, the iron(II) and iron(III) could not only be completely separated, but also be sensitively determined.     

Determination of quinolizidine alkaloids in traditional Chinese herbal drugs by nonaqueous capillary electrophoresis.

A rapid method for the determination of quinolizidine alkaloids by nonaqueous capillary electrophoresis was developed. A total of 10 alkaloids (matrine, sophocarpine, oxymatrine, oxysophocarpine, sophoridine, cytisine, sophoramine, aloperine, lehmannine and dauricine) could be easily separated within 18 min. A running buffer composed of 50 mM ammonium acetate, 10% tetrahydrofuran and 0.5% acetic acid in methanol was found to be the most suitable for this separation. Five of these alkaloids were selected for further studies. The linear calibration ranges were 2.51-50.1 microg/ml for sophoridine and sophocarpine, 2.71-54.2 microg/ml for matrine, 3.30-65.9 microg/ml for oxymatrine, and 3.10-62.0 microg/ml for oxysophocarpine. The recovery of the five alkaloids was 98.0-101.3% with relative standard deviations from 1.03 to 2.68% (n=5). The limits of detection for all 10 alkaloids were over the range 0.93-2.31 microg/ml. The method was successfully applied to the phytochemical analysis of alkaloid extracts from three commonly used traditional Chinese herbal drugs: Sophora flavescens Ait. (Kushen), S. alopecuroides L. (Kudouzi or Kugancao) and S. tonkinensis Gapnep (Shandougen).     

Determination of synthetic food dyes by capillary electrophoresis

A method for the determination of synthetic tar dyes used as food additives using capillary electrophoresis with photodiode-array detection was investigated. The dyes Erythrosine (R-3), Phloxine (R-104), Rose Bengal (R-105), Acid Red (R-106), Amaranth (R-2), New Coccine (R-102) and Allura Red AC (R-40) were separated on a capillary column (50 cm × 75 μm I.D.) and identified from the absorbance spectra of each peak. The electrophoresis buffer used was a mixture of 25 mM sodium phosphate buffer and 25 mM sodium borate buffer (1:1) (pH 8.0) containing 10 mM sodium dodecyl sulfate (SDS). Substitution of β-cyclodextrin for SDS in the electrolyte buffer was effective for the separation of R-2 and R-102. This modified method could be employed as an additional assay method for these two dyes.     

Determination of azide as the 3,5-dinitrobenzoyl derivative by capillary electrophoresis

A simple, rapid and reliable capillary electrophoresis method with a photodiode array detector was developed for determination of azide as the 3,5-dinitrobenzoyl derivative in drink samples fortified with sodium azide. Sample preparation was simple and rapid because no more than a simple dilution of samples is needed after quick derivatization. Separation was carried out using a buffer system comprising 25 mM phosphate buffer and 4 mM cetyltrimethylammonium hydroxide at pH 3.0. Methyl benzoate was selected as the internal standard (IS). This study investigated the influence of the concentration of phosphate buffer and electroosmotic flow (EOF) modifier, and the buffer pH on migration time and signal response. The optimized method made it possible to determine azide within 5 min. The limit of detection was determined to be 1.9 microg/ml with SIN > 3. The quantitation range was 6.5-323 microg/ml. By the method recoveries of azide in drink samples fortified with sodium azide were investigated. Mean recovery values ranged from 93.6 to 105.8% and results were satisfactory. In addition, no interference was observed in electropherograms of drink samples fortified with sodium azide. Thus, by this method, azide in drink samples can be determined rapidly with high recoveries and good selectivity despite extremely simple sample preparation.     

Analysis of vancomycin and related impurities by micellar electrokinetic capillary chromatography. Method development and validation

A fast and highly selective micellar electrokinetic capillary chromatography (MEKC) method for quantitative analysis of vancomycin and related impurities is described. Among the tested surfactants, cetyltrimethylammonium chloride (CTAC) offered the best selectivity. Another important parameter, which strongly influenced the selectivity, was buffer pH. It was found that the selectivity increased with buffer pH decreasing from 9 to 5. Using Tris-phosphate buffer containing CTAC, satisfactory separation could be obtained in the pH range from 5.0 to 5.5. Excellent repeatability in terms of migration time and peak area could be obtained when the capillary was carefully washed between two runs. In order to obtain optimal conditions and to evaluate the method robustness, a central composite experimental design was carried out. The optimal conditions were: 44 cm length of fused-silica capillary with 50 microm ID, 120 mM Tris-phosphate buffer (pH 5.2) containing 50 mM CTAC, -15 kV applied voltage, UV detection at 210 nm, and a column temperature of 25 degrees C. Under the optimal conditions, more than 20 peaks could be separated within 8 min. The method has a linearity range from 0.004 to 1.2 mg/ml (concentration of vancomycin B, active component). The limit of detection (LOD) and limit of quantitation (LOQ) were 0.4 microg/mL vancomycin, equivalent to 0.3 microg/mL vancomycin B (0.04%) and 1.1 microg/mL vancomycin, equivalent to 0.9 microg/mL vancomycin B (0.1%), respectively.     

Validated method for L-ornithine-L-aspartate analysis in human plasma by capillary electrophoresis

A rapid capillary electrophoresis method for routine determination of two amino acids, L-ornithine and L-aspartic acid, in human plasma is reported. The method runs automatically, requires a minimum of sample preparation and moreover includes no extensive extraction and no gradient or derivatization procedure. Analyses were performed on an uncoated silica capillary using buffer solution composed with 10 mM sodium tetraborate and 1 M sodium hydroxide (pH=10.0). A capillary electrophoresis P/ACE system equipped with UV detection (200 nm), an automatic injector, a fluid cooled cartridge and System Gold data station was used in this study. The total analysis time under these conditions was 8.0 min. The calibration curve was linear in the range 10-280 microg mL-1 for L-aspartic acid and 20-280 microg mL-1 for L-ornithine (for both amino acids, r=0.999). The method was validated by inaccuracy (bias) and precision (RSD) studies by analysing samples. The method was successfully applied to the quantitative determination of L-ornithine-L-aspartate in human plasma and could be useful for clinical and bioavailability investigations.     

Rapid separation of tetracycline derivatives and their main degradation products by capillary zone electrophoresis.

A mixture of five tetracycline (TC) derivatives: minocycline (MC), demeclocycline (DMCTC), doxycycline (DC), and sancycline (SC), as well as each TC derivative from its main degradation product were separated by capillary zone electrophoresis (CZE). The influence of the pH and the concentration and nature of the background electrolyte (BGE) on the separations was investigated. Ethylenediaminetetraacetic acid (EDTA; 1 mM) was used as additive in a 25 mM phosphate buffer (pH 2.3) because this BGE enabled the rapid separation of the TC derivatives and of each TC derivative from its respective degradation product in less than 6 min. After optimization of the separation conditions, the analytical characteristics of the method were investigated. The parameters involved were linearity, precision (repeatability and reproducibility), and limits of detection (LODs). LODs obtained for the five TC derivatives studied were about 3 microg/mL. Finally, the CZE method developed was applied to study the stability of TC derivatives and to analyze the TC derivative content in three different pharmaceutical preparations.