Development of a background electrolyte for the determination of inorganic cations in high ionic strength samples by capillary electrophoresis with indirect UV-absorption detection

In this study, a background electrolyte capable to separate and quantify inorganic cations in high ionic strength samples by UV-absorption indirect detection was designed. In this regard, the four most abundant monovalent and divalent cations in earth crust (K+ , Na+ , Ca+2, Mg+2) were selected as model compounds. A group of small carboxylic acids and, several toluidines and pyridines were evaluated as mild strength complexing agents and chromophoric probes, respectively. The optimized background electrolyte was composed of 200 mM 2,4,6-trimethylpyridine as the chromophoric probe, 250 mM lactic acid as the weak complexing agent and pH buffering reagent (adjusted to pH 4.5), and 5 % v/v methanol as organic solvent modifier. Based on a minimum number of components, it provided outstanding separation performance in less than 4 min in a wide linear dynamic range (10 - 2500 µg∙mL-1 ). Performances were contrasted against a reference method based on conductometric detection. Furthermore, studies of separation efficiency and peak shape were carried out at different analyte concentrations in high electric conductivity solutions. The herein developed method demonstrated exceptional features in terms of limits of detection (~10 µg∙mL-1 ), resolution, speed of analysis, sensitivity and peak capacity in high electric conductivity samples. Moreover, the method was successfully applied to high ionic strength samples such as rock digest, sea water, soy sauce and isotonic drinks.Fil: Lancioni, Carlina. Universidad Nacional de la Plata. Facultad de Cs.exactas. Laboratorio de Investigacion y Desarrollo de Metodos Analiticos. - Comision de Investigaciones Cientificas de la Provincia de Buenos Aires. Laboratorio de Investigacion y Desarrollo de Metodos Analiticos.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; ArgentinaFil: Aspromonte, Juan. Universidad Nacional de la Plata. Facultad de Cs.exactas. Laboratorio de Investigacion y Desarrollo de Metodos Analiticos. - Comision de Investigaciones Cientificas de la Provincia de Buenos Aires. Laboratorio de Investigacion y Desarrollo de Metodos Analiticos.; Argentina. Katholikie Universiteit Leuven; Bélgica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; ArgentinaFil: Tascon, Marcos. Universidad Nacional de San Martín. Instituto de Investigación e Ingeniería Ambiental. - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación e Ingeniería Ambiental; ArgentinaFil: Gagliardi, Leonardo Gabriel. Universidad Nacional de la Plata. Facultad de Cs.exactas. Laboratorio de Investigacion y Desarrollo de Metodos Analiticos. - Comision de Investigaciones Cientificas de la Provincia de Buenos Aires. Laboratorio de Investigacion y Desarrollo de Metodos Analiticos.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentin

A rapid and simple method for the determination of psychoactive alkaloids by CE-UV: application to Peganum Harmala seed infusions

The β-carboline alkaloids of the harmala (HAlks) group are compounds widely spread in many natural sources, but found at relatively high levels in some specific plants like Peganum harmala (Syrian rue) or Banisteriopsis caapi. HAlks are a reversible Mono Amino Oxidase type A Inhibitor (MAOI) and, as a consequence, these plants or their extracts can be used to produce psychotropic effects when are combined with psychotropic drugs based on amino groups. Since the occurrence and the levels of the HAlks in natural sources are subject to significant variability, more widespread use is not clinical but recreational or ritual, for example B. caapi is a known part of the Ayahuasca ritual mixture. The lack of simple methods to control the variable levels of these compounds in natural sources restricts the possibilities to dose in strict quantities and, as a consequence, limits its use with pharmacological or clinical purposes. In this work, we present a fast, simple, and robust method of quantifying simultaneously the six HAlksmore frequently found in plants, i.e., harmine, harmaline, harmol, harmalol, harmane, and norharmane, by capillary electrophoresis instruments equipped with the more common detector UV. The method is applied to analyze these HAlks in P. Harmala seeds infusion which is a frequent intake form for these HAlks. The method is validated in three different instruments in order to evaluate the transferability and to compare the performances between them. In this case, harmaline, harmine, and harmol were found in the infusion samples.Laboratorio de Investigación y Desarrollo de Métodos Analíticos (LIDMA

A high performance system to study the influence of temperature in on-line solid-phase extraction capillary electrophoresis

A novel high performance system to control the temperature of the microcartridge in on-line solid phase extraction capillary electrophoresis (SPE–CE) is introduced. The mini-device consists in a thermostatic bath that fits inside of the cassette of any commercial CE instrument, while its temperature is controlled from an external circuit of liquid connecting three differentwater baths. The circuits are controlled from a switchboard connected to an array of electrovalves that allow to rapidly alternate the water circulation through the mini-thermostatic-bath between temperatures from 5 to 90 ºC. The combination of the mini-device and the forced-air thermostatization system of the commercial CE instrument allows to optimize independently the temperature of the sample loading, the clean-up, the analyte elution and the electrophoretic separation steps. The system is used to study the effect of temperature on the C18-SPE–CE analysis of the opioid peptides, Dynorphin A (Dyn A), Endomorphin1 (END) and Met-enkephalin (MET), in both standard solutions and in spiked plasma samples. Extraction recoveries demonstrated to depend, with a non-monotonous trend, on the microcartridge temperature during the sample loading and became maximum at 60 ºC. Results prove the potential of temperature control to further enhance sensitivity in SPE–CE when analytes are thermally stable.Facultad de Ciencias ExactasLaboratorio de Investigación y Desarrollo de Métodos Analíticos (LIDMA)Centro de Investigación y Desarrollo en Tecnología de Pintura

Enantiomeric separations by capillary electrophoresis: Theoretical method to determine optimum chiral selector concentration

A method to optimize the ligand concentration [S] in the background electrolyte of capillary electrophoresis separations is presented. It is based on the use of a model which predicts apparent electrophoretic mobilities as a function of ligand concentration (expressed as p[S] = −log[S]). This model is employed to compose the expression of a recently proposed criterion to qualify separations in electrophoresis. Two strategies to find the optimum p[S], leading to the best separation of all compounds, are explained: 1.- a graphical method using a windows map depicting the single separation criteria between all possible combination of compounds by pairs, and 2.- an analytical method where an extended multicriterion optimization function is composed and optimum p[S] is found by mathematical maximization. The procedure is applied to a hard-to-separate model system: enantiomeric separations of racemic mixtures. 2-Hydroxypropyl-β-cyclodextrin was chosen as a model ligand, and four pharmaceutical drugs as model analytes. In order to demonstrate the performance of the procedure, results of electrophoretic separations obtained at p[S] found as optimum are compared with separations obtained at p[S] values slightly higher and lower than the optimum.Fil: Lancioni, Carlina. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Exactas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Keunchkarian, Sonia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Exactas; ArgentinaFil: Castells, Cecilia Beatriz Marta. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Exactas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Gagliardi, Leonardo Gabriel. Universidad Nacional de La Plata. Facultad de Ciencias Exactas; Argentina. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Determination of thermodynamic binding constants by affinity capillary electrophoresis

A strategy to study thermodynamic binding constants by affinity capillary electrophoresis (ACE) is presented. In order to simplify mathematical treatment, analogy with acid-base dissociation equilibrium is proposed: instead of ligand concentration [X], negative logarithm of ligand concentration (or activity), pX = -log[X], is used. On this base, and taking into account ionic activities, a general procedure for obtaining thermodynamic binding constants is proposed. In addition, the method provides electrophoretic mobilities of the free analyte and analyte-ligand complex, even when binding constants are low and thus, the complexed analyte fraction is also low. This is useful as a base to rationally analyze a diversity of situations, i.e., different mathematical dependencies are obtained when analytes and ligands with different charges are combined. Practical considerations are given for carrying out a full experimental design. Enantiomeric ACE separation based on the use of chiral selectors is addressed. 2-hydroxypropyl-β-cyclodextrin was chosen as a model ligand, and both enantiomeric forms of four pharmaceutical drugs (propranolol, pindolol, oxprenolol and homatropine methylbromide) were considered as model analytes. Practical aspects are detailed and thermodynamic binding constants as well as free and complexed analytes mobilities are determined.Fil: Lancioni, Carlina. Universidad Nacional de la Plata. Facultad de Cs.exactas. Laboratorio de Investigacion y Desarrollo de Metodos Analiticos. - Comision de Investigaciones Cientificas de la Provincia de Buenos Aires. Laboratorio de Investigacion y Desarrollo de Metodos Analiticos.; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Química. Grupo Cromatografía; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; ArgentinaFil: Keunchkarian, Sonia. Universidad Nacional de la Plata. Facultad de Cs.exactas. Laboratorio de Investigacion y Desarrollo de Metodos Analiticos. - Comision de Investigaciones Cientificas de la Provincia de Buenos Aires. Laboratorio de Investigacion y Desarrollo de Metodos Analiticos.; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Química. Grupo Cromatografía; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; ArgentinaFil: Castells, Cecilia Beatriz Marta. Universidad Nacional de la Plata. Facultad de Cs.exactas. Laboratorio de Investigacion y Desarrollo de Metodos Analiticos. - Comision de Investigaciones Cientificas de la Provincia de Buenos Aires. Laboratorio de Investigacion y Desarrollo de Metodos Analiticos.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; ArgentinaFil: Gagliardi, Leonardo Gabriel. Universidad Nacional de la Plata. Facultad de Cs.exactas. Laboratorio de Investigacion y Desarrollo de Metodos Analiticos. - Comision de Investigaciones Cientificas de la Provincia de Buenos Aires. Laboratorio de Investigacion y Desarrollo de Metodos Analiticos.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentin

Modeling retention and selectivity as a function of pH and column temperature in liquid chromatography

In reversed-phase liquid chromatography (RPLC), the retention of weak acids and bases is a sigmoidal function of the mobile-phase pH. Therefore, pH is a key chromatographic variable to optimize retention and selectivity. Furthermore, at an eluent pH close to the pKa of the solute, the dependence of ionization of the buffer and solute on temperature can be used to improve chromatographic separations involving ionizable solutes by an adequate handling of column temperature. In this paper, we derive a general equation for the prediction of the retentive behavior of ionizable compounds upon simultaneous changes in mobile-phase pH and column temperature. Four experiments, two limiting pH values and two temperatures, provide the input data that allow predictions in the whole range of these two variables, based on the thermodynamic fundamentals of the involved equilibria. Also, the study demonstrates the significant role that the choice of the buffer compound would have on selectivity factors in RPLC at temperatures higher than 25 °C. © 2006 American Chemical Society.Fil: Gagliardi, Leonardo Gabriel. Universidad de Barcelona; España. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Centro de Investigaciones en Tecnología de Pinturas. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones en Tecnología de Pinturas; ArgentinaFil: Castells, Cecilia Beatriz Marta. Universidad Nacional de La Plata; Argentina. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Centro de Investigaciones en Tecnología de Pinturas. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones en Tecnología de Pinturas; ArgentinaFil: Ràfols, Clara. Universidad de Barcelona; EspañaFil: Rosés, Martí. Universidad de Barcelona; EspañaFil: Bosch, Elisabeth. Universidad de Barcelona; Españ

Síntesis y caracterización de polímeros de coordinación de lantánidos: potencial aplicación a la determinación de contaminantes ambientales mediante metodologías fotoluminiscentes

En el presente trabajo, se llevó a cabo la síntesis del polímero de coordinación TbBTC a partir de ácido bencen-1,3,5-tricarboxílico (H3BTC) y nitrato de terbio hexahidratado en medio agua/etanol. Se obtuvieron nanoestructuras alargadas (nanobelts) que exhiben una emisión fotoluminiscente característica del terbio, con tiempos de vida del orden de los mili-microsegundos. Por otra parte, se incluyó al TbBTC en matrices poliméricas sol-gel funcionalizadas con distintos grupos, utilizando tetraetoxisilano (TEOS) como entrecruzante y dos precursores funcionales (3-aminopropil trietoxisilano, APTES; 3-mercaptopropil trimetoxisilano, MPTS), obteniéndose un material híbrido fotoluminiscente. Los materiales desarrollados se caracterizaron morfológicamente por microscopía electrónica de barrido (SEM); se estudiaron sus propiedades ópticas, en suspensión y en estado sólido, y cómo éstas se ven afectadas por la presencia de ciertos analitos de interés a partir de ensayos de contacto en solución acuosa.Laboratorio de Investigación y Desarrollo de Métodos Analítico

Development of a background electrolyte for the determination of inorganic cations in high ionic strength samples by capillary electrophoresis with indirect UV-absorption detection

In this study, a background electrolyte capable to separate and quantify inorganic cations in high ionic strength samples by UV-absorption indirect detection was designed. In this regard, the four most abundant monovalent and divalent cations in earth crust (K+, Na+, Ca+2, Mg+2) were selected as model compounds. A group of small carboxylic acids and, several toluidines and pyridines were evaluated as mild strength complexing agents and chromophoric probes, respectively. The optimized background electrolyte was composed of 200 mM 2,4,6-trimethylpyridine as the chromophoric probe, 250 mM lactic acid as the weak complexing agent and pH buffering reagent (adjusted to pH 4.5), and 5% v/v methanol as organic solvent modifier. Based on a minimum number of components, it provided outstanding separation performance in less than 4 min in a wide linear dynamic range (10 - 2500 µg·mL−1). Performances were contrasted against a reference method based on conductometric detection. Furthermore, studies of separation efficiency and peak shape were carried out at different analyte concentrations in high electric conductivity solutions. The herein developed method demonstrated exceptional features in terms of limits of detection (~10 µg·mL−1), resolution, speed of analysis, sensitivity and peak capacity in high electric conductivity samples. Moreover, the method was successfully applied to high ionic strength samples such as rock digest, sea water, soy sauce and isotonic drinks.Laboratorio de Investigación y Desarrollo de Métodos Analíticos (LIDMA

Fast determination of harmala alkaloids in edible algae by capillary electrophoresis mass spectrometry

The use of algae as a foodstuff is rapidly expanding worldwide from the East Asian countries, where they are also used for medical care. Harmala alkaloids (HAlk) are a family of bioactive compounds found in the extracts of some plants, including wakame (Undaria pinnatifida), an edible marine invasive algae. HAlks are based on a characteristic βcarboline structure with at least one amino ionizable group. In this work, we report the successful separation of a mixture of six HAlks (harmine, harmaline, harmol, harmalol, harmane, and norharmane) by capillary electrophoresis ion-trap mass spectrometry (CE-IT-MS) in less than 8 min. Optimum separation in fused-silica capillaries and detection sensitivity in positive-ion mode were achieved using a background electrolyte (BGE) with 25 mmol L⁻¹ ammonium acetate (pH 7.8) and 10 % (v/v) methanol, and a sheath liquid with 60:40 (v/v) isopropanol–water and 0.05 % (v/v) formic acid. The separation method was validated in terms of linearity, limits of detection and quantification, repeatability, and reproducibility. Later, a sample pretreatment was carefully optimized to determine HAlks in commercial wakame samples with excellent recovery and repeatability. For the complex wakame extracts, the MS–MS fragmentation patterns of the different HAlks were useful to ensure a reliable identification. The complete procedure was validated using the standardaddition calibration method, determining matrix effects on the studied compounds. Harmalol, harmine, and harmaline were naturally present in the samples and were quantified at very low concentrations, ranging from 7 to 24 μg kg⁻¹ dry algae.Facultad de Ciencias ExactasLaboratorio de Investigación y Desarrollo de Métodos Analíticos (LIDMA)Centro de Investigación y Desarrollo en Tecnología de Pintura

Determination of thermodynamic binding constants by affinity capillary electrophoresis

A strategy to study thermodynamic binding constants by affinity capillary electrophoresis (ACE) is presented. In order to simplify mathematical treatment, analogy with acid-base dissociation equilibrium is proposed: instead of ligand concentration [X], negative logarithm of ligand concentration (or activity), pX = -log[X], is used. On this base, and taking into account ionic activities, a general procedure for obtaining thermodynamic binding constants is proposed. In addition, the method provides electrophoretic mobilities of the free analyte and analyte-ligand complex, even when binding constants are low and thus, the complexed analyte fraction is also low. This is useful as a base to rationally analyze a diversity of situations, i.e., different mathematical dependencies are obtained when analytes and ligands with different charges are combined. Practical considerations are given for carrying out a full experimental design. Enantiomeric ACE separation based on the use of chiral selectors is addressed. 2-hydroxypropyl-β-cyclodextrin was chosen as a model ligand, and both enantiomeric forms of four pharmaceutical drugs (propranolol, pindolol, oxprenolol and homatropine methylbromide) were considered as model analytes. Practical aspects are detailed and thermodynamic binding constants as well as free and complexed analytes mobilities are determined.Laboratorio de Investigación y Desarrollo de Métodos Analíticos (LIDMA