Determination of Picoxystrobin and Pyraclostrobin by MEKC with On-Line Analyte Concentration

Micellar electrokinetic capillary chromatography was used for the determination of picoxystrobin and pyraclostrobin. The background electrolyte consisted of borate buffer (40 mmol L⁻¹ pH 8.5), SDS (30 mmol L⁻¹) and acetonitrile (15% in volume). Runs were made at 25 °C with 25 kV applied potential. The developed method was applied to analyte fortified urine samples. On-line analyte concentration, combined with a capillary of a longer optical path length, allowed limits of quantification of 8.6 × 10⁻⁸ mol L⁻¹ for picoxystrobin and 1.8 × 10⁻⁷ mol L⁻¹ for pyraclostrobin.

     

Determination of Fatty Acids (C1 – C10) from Bryophytes and Pteridophytes

A simple and mild method for the determination of fatty acids (C₁ – C₁₀) based on a condensation reaction using 7-aminonaphthalene-1,3-disulfonic acid (ANDSA) as labeling reagent with capillary zone electrophoresis has been developed. The detection was performed with a diode array detector at 254 nm. A 58.5 cm × 50 μm i.d. (50 cm effective length) untreated fused-silica capillary was used. To optimize the separation conditions, the background electrolyte concentration, column temperature, voltage and other factors were evaluated. The optimal separation conditions were as follows: 30 mmol L⁻¹ borate buffer (pH 9.5), 15 mmol L⁻¹ β-CD, temperature at 20 °C, pressure 50 mbar and injection time 8 s. Under the established conditions, 10 fatty acid derivatives could be well-separated within 17 min. The linearity was in the range of 0.07–5.0 μmol L⁻¹. Detection limits (at a signal-to-noise ratio of 3) were in the range of 0.027–0.042 μmol L⁻¹. The fatty acids from the extracted Funaria Hedw. and Selaginella samples were determined with satisfactory results.

     

GC–MS Method Development for the Analyses of Thiophenes from Solvent Extracts of Tagetes patula L

A method for isotachophoretic determination of potassium and ammonium cations in fertilizers and silage was developed. A capillary of 0.4 mm i.d. and 100 mm effective length made of fluorinated ethylene–propylene copolymer was filled with an electrolyte system consisting of 10 mmol L⁻¹ RbOH + 0.1% (w/v) hydroxyethylcellulose, adjusted to pH 9.0 with l-histidine (leading electrolyte) and 10 mmol L⁻¹ lithium citrate (terminating electrolyte). Using contactless conductivity detection, the calibration curves in the tested concentration range up to 0.5 mmol L⁻¹ were linear for both cations. The concentration detection limits for potassium and ammonium were 2.9 and 2.7 μmol L⁻¹, respectively. RSD values of step lengths (n = 6) were 1.3% for potassium and 1.5% for ammonium. The separation time was about 20 min. Similar results were obtained with cesium cation used as the leading ion, however, in the system with rubidium better resolution of other cations present in tested matrices was reached. The elaborated method is simple to perform, sufficiently sensitive and accurate and can be recommended as an alternative procedure to the methods used so far for the determination of potassium and ammonium.

     

CE Profiling of Organic Acids in Distilled Alcohol Beverages Using Pattern Recognition Analysis

The objective of this work has been to assess the potential of capillary isotachophoretic organic acids profiling using multivariate statistical methods to classify brandy samples and wine distillate samples. The leading electrolyte was 10 mmol L⁻¹ hydrochloric acid including 0.1% methylhydroxylethylcellulose adjusted with β-alanine to pH 2.9. The terminating electrolyte was 5 mmol L⁻¹ acetic acid. Principal component analysis, cluster analysis, and linear discriminant analysis were used for the classification of beverages. The results show that for the 12 acids analysed, 98.57% of the total variance is extracted by the six principal components (PC). After performing backward linear discriminant analysis, a classification function was obtained containing four variables: formic (PC2-loadings: 0.989), lactic (PC1-loadings: 0.886), malic (PC1-loadings: 0.989) and oxalic (PC2-loadings: 0.777) acids, which provide 100.0% correct classification of brandies and wine distillates.

     

A Rapid Determination of Taurine in Human Plasma by LC

Taurine is an amino acid which is not incorporated into proteins but found in the cytosol of many mammalian cells, in high concentrations (2–30 mM). Increase in plasma taurine concentration has already been reported after surgical trauma, X-radiation, muscle necrosis, carbon tetrachloride-induced liver damage, and paracetamol overdose. Plasma taurine concentration was measured using LC with fluorescence detection following derivatization by o-phtalaldehyde plus 3-mercapto-propionic acid and α-aminobutyric acid as internal standard. Under these conditions the retention time of taurine was 10 min. This method was sensitive enough, to quantify 150 pg mL⁻¹ and detect 50 pg mL⁻¹ of taurine ranging normally between 65 and 179 mmol L⁻¹ (8–22 μg mL⁻¹). The validated method allowed simple determination of human plasma taurine in pharmacokinetic and biomarker studies.

     

Simultaneous Determination of Toxic Alkaloids in Blood and Urine by HPLC–ESI–MS/MS

A sensitive and selective method for simultaneous determination of 29 toxic alkaloids in human blood and 31 in urine using high-performance liquid chromatography–electrospray ionization-tandem mass spectrometry was developed and validated. The samples were diluted with 0.1 mol L⁻¹ HCl, and the target alkaloids were purified by solid phase extraction. The separation of 31 alkaloids was carried out on a C18 column using a gradient mobile phase with 10 mmol L⁻¹ ammonium formate in water with 0.1% formic acid and methanol at the rate of 0.25 mL min⁻¹. The triple-quadrupole mass spectrometer equipped with an electrospray source in the positive mode was set up in the dynamic multiple reactions monitoring mode (dynamic MRM) to detect the ion transitions of 31 alkaloids. The calibration curves were linear over a range of 0.5–400, 1–400, or 4–400 μg L⁻¹ for target alkaloids in human blood and urine, and the correlation coefficients (r ²) was higher than 0.9943. The limit of determination and limit of quantification were 0.2–1 and 0.5–4 μg L⁻¹ for blood and urine, respectively. The only exceptions were sanguinarine and chelerythrine in human blood. All the target alkaloids were stable under the test condition. In addition, the solvent effect and reconstituted solution were investigated. The method was validated and proved to be accurate and precise over the studied concentrations and suitable for poisoning diagnosis and forensic toxicology.

     

Pulsed amperometric detection (PAD) of diuretic drugs in herbal formulations using a gold electrode following ion-pair chromatographic separation

Herein, we present a new method based on separation by ion-pair chromatography with pulsed amperometric detection for evaluating various diuretics, including hydrochlorothiazide, chlorthalidone, furosemide, and amiloride, which are adulterants in herbal-based pharmaceutical formulations. The amperometric detection cycle (time?=?2 s) was performed at a gold electrode by applying a detection potential (E1) of +800 mV for 0.4 s and an oxidation potential (E2) of +1,000 mV for 0.40 s, followed by a reduction potential (E3) of ?200 mV for 1.20 s. The mobile phase for separating the diuretics was composed of 5 mmol L^?1 phosphate buffer and 0.3 mmol L^?1 sodium dodecyl sulfate in 50 % (v/v) methanol (pH 4.5). This method enabled the quantification of the drugs at low concentrations (i.e., 0.08 mg/capsule for hydrochlorothiazide, 0.01 mg/capsule for chlorthalidone, and 0.007 mg/capsule for furosemide). Twenty-six herbal formulations were analyzed, and eight samples (30.8 %) were found to contain diuretics that were added to the final composition (declared or not).     

Sensitive Analysis of Wilforidine in Human Plasma by LC-APCI-MS/MS

Wilforidine is a potentially efficient medicine to cure autoimmune diseases. In this paper, a sensitive and selective liquid chromatographic method coupled with atmospheric -pressure chemical ionization mass spectrometry (LC–APCI–MS/MS) has been developed for quantification of wilforidine in human plasma. Samples were deproteinized with acetonitrile and cleaned by solid-phase extraction. The chromatographic separation was performed on an analytical RRHD C₁₈ column (50 × 2.1 mm) using ammonium acetate solution (10.0 mmol L⁻¹)/acetonitrile (30/70, v/v) as the mobile phase at a flow rate of 0.7 mL min⁻¹. Detection was carried out by the positive multiple reaction monitoring mode with transitions of m/z 780 → 684 for wilforidine, and 646 → 586 for aconitine (internal standard), respectively. The calibration curve was linear (r = 0.9991) in the concentration range of 0.5–100.0 μg L⁻¹ with a lower limit of quantification of 0.5 μg L⁻¹ in plasma. Intra- and inter-day relative standard deviations were less than 6.8 and 13.1 %, respectively, and the recoveries were between 88.0 and 96.0 %. This accurate and highly specific assay provides a useful method for evaluating the pharmacokinetics of wilforidine in human plasma.

     

Simultaneous determination of first-line anti-tuberculosis drugs by capillary zone electrophoresis using direct UV detection

An alternative methodology for simultaneous analysis of ethambutol, isoniazid, rifampicin and pyrazinamide in pharmaceutical formulations by capillary zone electrophoresis under UV direct detection with an analysis time of 8.0 min is proposed. Background running was based on the effective mobility curve of the analytes and an optimum separation condition was achieved using a 3³ Box-Behnken design, with Brij 35, Cu²⁺ and acetic acid/sodium acetate buffer as factors. An electrolyte consisting of 50.0 mmol L⁻¹ of acetic acid/sodium acetate buffer, 12.5 mmol L⁻¹ of CuSO₄, and standard and sample solutions prepared in 2.00 mmol L⁻¹ of Brij 35 and 12.5 mmol L⁻¹ of CuSO₄ were optimized. After evaluating validation parameters, the method was successfully applied to the analysis of samples in the form of tablets and sachets.     

A fast,highly selective and sensitive dansyl-based fluorescent sensor for copper (II) ions and its imaging application in living cells

Since the copper ions (Cu²⁺) play a fatal role in many foundational physiological processes, it is important to develop a simple, highly sensitive and selective sensor for Cu²⁺ detection in living systems. Herein, an intramolecular charge transfer (ICT) and dansyl-based fluorescent chemosensor 1 was designed, synthesized and characterized for the sensitive and selective quantification of Cu²⁺. It exhibited remarkable fluorescence quenching upon addition of Cu²⁺ over other selected metal ions, attributed to the complex formation between 1 and Cu²⁺ with the association constant 6.7 × 10⁵ M^?1. The sensor 1 showed a fast and linear response towards Cu²⁺ in the concentration range from 0 to 12.5 × 10^?6 mol L^?1 with the detection limit of 2.5 × 10^?7 mol L^?1. This detection could be carried out in a wide pH range of 5.0–14. Furthermore, sensor 1 can be used for detecting Cu²⁺ in living cells.     

Rapid Determination of Trifluoromethanesulfonate and p-Toluenesulfonate by Ion-Pair Chromatography Using a Reversed-Phase Silica-Based Monolithic Column: Application to the Analysis of Ionic Liquids

A method of ion-pair chromatography was developed on a reversed-phase silica-based monolithic column for the fast and simultaneous determination of trifluoromethanesulfonate (CF₃SO₃ ⁻) and p-toluenesulfonate (C₇H₇SO₃ ⁻). The analysis was performed using a mobile phase of tetrabutylammonium hydroxide + citric acid + acetonitrile on the Chromolith Speed ROD RP-18e column with direct conductivity detection. The effects of the eluent, column temperature and flow rate on the retention of the anions were investigated. The experimental phenomenon was discussed according to hydrophobic interaction and ion-exchange mechanism in the separation. The optimized chromatographic conditions were selected. The optimized eluent for the separation consisted of 0.2 mmol L⁻¹ tetrabutylammonium hydroxide + 0.10 mmol L⁻¹ citric acid + 9% acetonitrile (pH 5.5). The flow rate was set at 6.0 mL min⁻¹. The column temperature was 25 °C. Under the optimal conditions, the better separation of CF₃SO₃ ⁻ and C₇H₇SO₃ ⁻ was achieved without any interference by other anions (Cl⁻, Br⁻, I⁻, NO₃ ⁻, SO₄ ²⁻, ClO₃ ⁻, BF₄ ⁻ and PF₆ ⁻). The detection limit (S/N = 3) was 0.28 and 0.71 mg L⁻¹ for CF₃SO₃ ⁻ and C₇H₇SO₃ ⁻, respectively. The method has been applied to the determination of CF₃SO₃ ⁻ and C₇H₇SO₃ ⁻ in ionic liquids. The spiked recoveries of CF₃SO₃ ⁻ and C₇H₇SO₃ ⁻ were 101.1 and 100.2%, respectively.

     

Optimization and Validation of a Capillary MEKC Method for Determination of Proteins in Urine

Proteinuria, i.e. increased excretion of proteins in urine, is a common sign indicating renal or urinary tract diseases. In this study, a fast and simple procedure for urine sample preparation and capillary micellar electrokinetic chromatographic analysis is presented, without any sample pretreatment prior to the analysis. The developed MEKC method was employed for simultaneous determination of albumin (ALB), haemoglobin (HGB), and myoglobin (MYO) in human urine samples obtained from patients with diagnosed proteinuria. Optimum conditions for detection and separation of ALB, HGB, and MYO are 50 mmol L⁻¹ borate buffer containing 20 mmol L⁻¹ SDS (pH 9.3), injection 40 mbar × 20 s, voltage 25 kV, temperature 30 °C, and detection wavelength 200 nm. The method was shown to be specific, accurate, linear (correlation coefficients r ² > 0.99), and precise (RSD below 3.75 and 7.23% for migration time and peak area, respectively). Multi-variable-at-a-time (MVAT) approach for robustness testing shows no significant variations in accuracy, specificity, and precision as RSD values were lower than 5 and 10% for migration time and peak area, respectively. The presented method is applicable for routine analyses of urine samples as a screening method for patients with excess ALB, HGB, and MYO.

     

Determination of DNA Methylation and Hydroxymethylation Levels in Biological Samples by Field-Amplified Sample Injection-Capillary Zone Electrophoresis with UV Detection

In this study, a method of field-amplified sample injection coupled with capillary zone electrophoresis with ultraviolet detection was established for evaluation of DNA methylation and hydroxymethylation levels in biological materials. By modifying an existing method, the separation of cytosine (C), 5-methylcytosine (5-mC) and 5-hydroxymethylcytosine (5-hmC) was performed on an uncoated capillary column (40 cm × 75 μm I.D.) using 300 mmol L⁻¹ tris solution (pH 2.90) as running buffer and detected at 280 nm. The detection limits (S/N = 3) of the method were 0.004 ng mL⁻¹ for cytosine (C), 0.01 ng mL⁻¹ for 5-methylcytosine (5-mC), and 0.02 ng mL⁻¹ for 5-hydroxymethylcytosine (5-hmC). The proposed method has been successfully applied to the evaluation of DNA methylation and hydroxymethylation levels of blood samples from 15 hepatocellular carcinoma patients and 5 liver cirrhosis patients and liver tissues from 50 pairs of tumor and matched tumor-adjacent samples.

     

Determination of Lipoic Acid in Biological Samples with Acetonitrile–Salt Stacking Method in CE

An acetonitrile–salt stacking method was established for the assay of lipoic acid (LA) in biological samples. Samples were deproteinized with acetonitrile at a final concentration of 60 % (v/v) and then injected hydrodynamically at 3.45 × 10³ Pa for 180.0 s. The optimum background electrolyte was found to be 90.0 mmol L^?1 pH 9.1 borate buffer. LA could be detected within 35 min at +7.0 kV with satisfactory repeatability (relative standard deviations, RSDs, of migration times and peak areas were both below 10 % for intraday and interday; n = 6/9) and a relatively low limit of detection of ca. 0.5 μmol L^?1.     

Electroanalytical Characteristics of Lercanidipine and its Voltammetric Determination in Pharmaceuticals and Human Serum on Boron-Doped Diamond Electrode

The electrooxidative behavior and determination of lercanidipine (LRC) were investigated in aqueous acetonitrile medium at a boron-doped diamond electrode using voltammetric techniques. The LRC in selected supporting electrolyte presents a well-defined anodic response at 0.944 V, studied by the proposed method. The linear response was obtained in the ranges of 4 × 10^?6 to 2 × 10^?4 mmol L^?1 range in 0.5 mmol L^?1 sulfuric acid supporting electrolyte and 1 × 10^?5 to 8 × 10^?5 mmol L^?1 range in spiked serum sample for square wave voltammetric technique. No electroactive interferences from the excipients and endogenous substances were found in the pharmaceutical dosage form and in the biological sample, respectively.     

A Simple Method for Determination of Homologue Imidazolium Cations in Ionic Liquids Using IC with Direct Conductivity Detection

An investigation was carried out into the fast determination of five homologue imidazolium cations in ionic liquids by ion chromatography using a cation-exchange column and direct conductivity detection. Ethylenediamine, complex organic acid (citric acid, oxalic acid and tartaric acid) and organic modifiers (acetonitrile) were used as mobile phase. The influences of the eluent types, eluent concentration, eluent pH and column temperature on separation of the cations were discussed. Simultaneous separation and determination of the five homologue imidazolium cations in ionic liquids were achieved under an optimum condition. The optimized mobile phase was consisted of 0.25 mmol L^?1 ethylenediamine + 0.5 mmol L^?1 citric acid + 3% acetonitrile (v/v) (pH 4.1), set at a flow rate of 1.0 mL min^?1. The column temperature was 40 °C and detection limits were obtained in the range of 1.1–45.6 mg L^?1. The relative standard deviations of the chromatographic peak areas for the cations were <3.0% (n = 5). This method was successfully applied to separate imidazolium cations in ionic liquids produced by organic synthesis. The recoveries of spiked components were 92.5–101.9%.     

Contribution of CuxO distribution,shape and ratio on TiO2 nanotubes to improve methanol production from CO2 photoelectroreduction

Many studies are focused on the development of materials for converting carbon dioxide into multicarbon oxygenates such as methanol and ethanol, because of their higher energy density and wider applicability. In this work, TiO₂ nanotubes (NT/TiO₂) were modified with Cu_xO nanoparticles in order to investigate the contribution of different ratio of Cu₂O/CuO and its distribution over NT/TiO₂ for CO₂ photoelectro-conversion to methanol. The photoelectrodes were built by anodization process to obtain NT/TiO₂ layer, and the decoration with Cu_xO hybrid system was carried out by electrodeposition process, using Na₂SO₄ or acid lactic as electrolyte, followed by annealing at different temperatures. X-ray photoelectron spectroscopy analysis revealed the predominance of Cu⁺¹ and Cu⁺² at 150 °C and 300 °C, respectively. X-ray diffraction and scanning electron microscopy indicated that under lactic acid solution, the oxide nanoparticles exhibited small size, cubic shape, and uniform distribution on the nanotube wall. While under Na₂SO₄ electrolyte, large nanoparticles with two different morphologies, octahedral and cubic shapes, were deposited on the top of the nanotubes. All modified electrodes converted CO₂ in methanol in different quantities, identified by gas chromatograph. However, the NT/TiO₂ modified with CuO/Cu₂O (80:20) nanoparticles using lactic acid as electrolyte showed better performance in the CO₂ reduction to methanol (0.11 mmol L⁻¹) in relation to the other electrodes. In all cases, a blend among the structures and nanoparticle morphologies were achieved and essential to create new site of reactions what improved the use of light irradiation, minimization of charge recombination rate and promoted high selectivity of products.

     

Simultaneous Analysis of Sugars and Polyphenols in Tobacco by CZE with Amperometric Detection Based on a Cu2O/MWCNT-COOH Composite Electrode

A composite electrode was fabricated from Cu₂O powder, carboxyl-functionalized multi-wall carbon nanotubes (MWCNT-COOH), and paraffin oil in the proportions 51:17:32 (w/w). This composite electrode was used for amperometric detection (CZE–AD) in simultaneous capillary zone electrophoretic analysis of chlorogenic acid, rutin, sucrose, glucose, mannose, and fructose in tobacco samples. Under the optimum conditions, the six analytes could be separated in 100 mmol L^?1 NaOH buffer within 30 min. Good linearity was achieved in the range 1 × 10^?7–1 × 10^?4 mol L^?1 for the two polyphenols and 5 × 10^?6–1 × 10^?3 mol L^?1 for the four sugars. The detection limits (S/N = 3) for the polyphenols and sugars were as low as 10^?8 mol L^?1 and 10^?6 mol L^?1, respectively.     

Influence of sodium addition on taurine adduct formation generated in acetic acid/acetate salt buffer applied in LC–MS/MS analysis

To investigate the influence of sodium ion addition on analyte adduct formation generated in acetic acid/acetate salt buffer appropriate infusion experiments of a 1 and 10 μg/mL taurine solution prepared both in methanol with a 1 mmol L^?1/10 mmol L^?1 CH₃COONH₄/CH₃COOH buffer and with a 1 mmol L^?1/10 mmol L^?1 CH₃COONa/CH₃COOH buffer were performed. The results achieved revealed that sodium ion concentration has a relevant influence on taurine adduct formation in the negative electrospray ionisation mode since depending on conditions applied different analyte adducts are favored. Sodium ions which originate from the glassware/chemicals make an effective detection of taurine dimer adducts possible, but taurine adducts with sodium acetate are not formed effectively. On the other side sodium ion addition enables an effective taurine adduct formation with sodium acetate, but taurine dimer based adducts can be observed only at higher analyte concentrations.     

Application of Double Deposition and Stripping Steps System for Minimization of Interferences of Peaks’ Overlapping in Anodic Stripping Voltammetry

An anodic stripping voltammetric procedure for the determination of bismuth in the presence of excess of Cu²⁺ ions at two ex situ plated gold film electrodes was described. The procedure is based on utilization of two deposition and two stripping steps system. The presented procedure ensures increasing the sensitivity of Bi³⁺ determination and minimization of interferences related to peaks’ overlapping. The calibration graph for bismuth determination was linear from 2.5×10⁻⁹ to 2×10⁻⁸ mol L⁻¹ for deposition time of 300 s at both working electrodes while detection limit was 7.7×10⁻¹⁰ mol L⁻¹.