Eletrocromatografia capilar: contextualização, estado da arte e perspectivas

Capillary electrochromatography (CEC) is a separation technique in which the mobile phase flow is based on the application of a voltage across a packed capillary, which generates an electroosmotic flow that transports the analytes along the capillary toward the detector. As it combines the separation mechanisms of high-performance liquid chromatography (HPLC) and of capillary electrophoresis (CE), CEC can be considered a hybrid of HPLC and CE. This review presents some fundamental aspects of CEC and is focused on the instrumental advances of the technique, such as column technology, operation modes and detection systems, presenting recent papers on these topics and some applications and perspectives about CEC.

Fotodegradación heterogénea de bisfenol A en agua con dióxido de titanio

Bisphenol A (BPA) is a monomer used in epoxy resin and polycarbonate manufacture. This molecule is considered as an endocrine disruptor that causes different diseases. The human exposition to this non biodegrable substance is increasing in the time; in particular, water is contaminated by industrial remainder flow. In this article heterogeneous photo degradation of a solution of BPA in water solution using a catalytic photo reactor with UV light and titanium dioxide (TiO2) was evaluated. High performance liquid chromatography (HPLC) was used to analyze the photo degradation of BPA solutions. The influence of titanium dioxide amount, BPA concentration, reaction temperature and the catalyst state like suspension and immobilized were also determinated. The highest elimination of BPA was 83.2%, in 240 min, beginning with 0.05 mM of BPA and 100 mg/L of TiO2 in suspension.

Aplicações de cromatografia líquida de alta eficiência para o estudo de poluentes orgânicos emergentes

Emerging organic pollutants (EOP) include many environmental contaminants based on commercial products such as pharmaceuticals, personal care products, detergents, gasoline, polymers, etc. EOP may be candidates for future regulation as they offer potential risk to environmental and human health due to their continual entrance into the environment and to the fact that even the most modern wastewater treatment plants are not able to totally transform / remove these compounds. High performance liquid chromatography is recommended to separate emerging organic pollutants with characteristics of high polarity and low volatility, especially pharmaceuticals, from environmental matrices.

Desenvolvimento e validação de método analítico por CLAE para a quantificação simultânea de dipropionato de betametasona e fosfato sódico de betametasona em suspensão injetável

A reversed-phase HPLC method was developed and validated to separate and simultaneously quantify the association of betamethasone sodium phosphate (BP) and betamethasone dipropionate (BD) in injectable suspensions. Chromatographic conditions were ternary gradient elution at 1.6 mL/min on a C18 column with 254 nm. The linearity of the method was established in the range 120 to 280 mg/mL BD, and 48 to 112 mg/mL BP. The RSD of intermediate precision of the method was <1% and recoveries were 99-101% for both drugs. The method proved selective, linear, precise, accurate and robust for quantifying BP and BD in commercial injectable suspensions.

Biodegradação de paclobutrazol por Pseudomonas spp. em sistemas de solo saturados

Paclobutrazol is growth regulator of plants that has low mobility in soil and therefore has accumulated. The objective of this study was to investigate the paclobutrazol biodegradation in two soils from the São Francisco River Valley. The biodegradation experiments were conducted in batch using paclobutrazol and paclobutrazol added glycerol. The experiments were performed in sterile and nonsterile conditions using a mixed culture of Pseudomonas. The concentration of paclobutrazol was determined by high performance liquid chromatography. The biodegradation reached 43% in 14 days of experiments with only paclobutrazol and 70% in 28 days of experiments that contained glycerol and paclobutrazol.

Desenvolvimento e validação de métodos analíticos para determinação de ácido glicirrízico, ácido salicílico e cafeína em nanopartículas de quitosana e alginato

The present work consists of the development and validation of analytical method for evaluation of glycyrrhizic acid, salicylic acid, and caffeine in chitosan-alginate nanoparticles by high performance liquid chromatography. Method validation investigated parameters such as linearity, precision, accuracy, robustness and specificity, which gave results within the acceptable range. The methods were applied to nanoparticles suspensions containing the drugs and were able to determine the entrapment efficiency successfully. The best entrapment efficiency was achieved with the glycyrrhizic acid (95.4%).

Preparação, caracterização e aplicação de fases estacionárias tipo C8 modificadas por óxidos metálicos para cromatografia líquida

This work describes three C8-stationary phases for high performance liquid chromatography based on silica metallized with ZrO2, TiO2 or Al2O3 layers, having poly(methyloctylsiloxane) immobilized onto their surfaces. The stationary phases were characterized using XRF, XAS, FTIR, SEM and elemental analysis to determine the physical characteristics of the oxide and polysiloxane layers formed on the surfaces and chromatographically to evaluate the separation parameters. The results show the changes on the silica surface and allowed proposing a structure for the oxide layer, being observed tetrahedral and octahedral structures, what is completely new in the literature. The formation of a homogeneous layer of metallic oxide (TiO2 and ZrO2) was observed on the silica. The C8-titanized and C8-aluminized stationary phases presented good chromatographic performances, with good values of asymmetry and efficiency. All stationary phase presented few loss of the polymeric layer after the HPLC, indicating that this layer is well attached on the metalized support.

Determinação simultânea de carbamazepina, fenitoína e fenobarbital em sangue seco em papel por cromatografia líquida de alta eficiência

Carbamazepine, phenobarbital and phenytoin were determined in dried blood spots (DBS) by high performance liquid chromatography, after extraction of 8 mm DBS using a mixture of acetonitrile and methanol. Analytes were separated by reversed-phase chromatography, with a run time of 17 minutes. Intra-assay and inter-assay precisions were in the 5.3 to 8.4% and 3.3 to 5.2% ranges, respectively. Accuracy was in the 98.8 to 104.3% range. The method had sensitivity to detect all analytes at levels below minimum therapeutic concentrations. The analytes were stable at 4 ºC and room temperature for up to 12 days and at 45 ºC for 9 days. The method was applied to 14 paired clinical samples of blood serum and DBS.

CROMATOGRAFIA POR INTERAÇÕES HIDROFILICAS (HILIC): ESTADO DA ARTE E APLICAÇÕES

Hydrophilic interaction liquid chromatography (HILIC) has been gaining increased attention for its effective separation of highly polar compounds, which include carbohydrates, amino acids, pharmaceutical compounds, proteins, glycoproteins, nucleosides, etc. Polar compounds are usually poorly retained on reverse-phase liquid chromatography (RP-HPLC) columns or have poor solubility in the apolar mobile phase of normal-phase high performance liquid chromatography (NP-HPLC). Since HILIC uses organic solvents such as ACN or MeOH ( > 70%), also used in RP-HPLC and polar stationary phases similar to NP-HPLC (bare silica, diol, amino, amide, saccharide, zwitterionic stationary phases, etc.), it represents a hybrid of the two separation modes. The high organic content in the MP leads to good compatibility with mass spectrometry (MS), increasing the detectivity. This review describes the fundamentals of HILIC and highlights some interesting applications.

Amperometric biosensor for ascorbic acid

A L-ascorbic acid biosensor based on ascorbate oxidase has been developed. The enzyme was extracted from the mesocarp of cucumber (Cucumis sativus) by using 0.05 mol L-1 phosphate buffer, pH 5.8 containing 0.5 mol L-1 NaCl. After the dialysis versus phosphate buffer 0.05 mol L-1 pH 5.8, the enzyme was immobilized onto nylon net through glutaraldehyde covalent bond. The membrane was coupled to an O2 electrode and the yielding reaction monitored by oxygen depletion at -600 mV using flow injection analysis optimized to 0.1 mol L-1 phosphate buffer pH 5.8, as the carrier solution and flow-rate of 0.5 mL min-1. The ascorbic acid calibration curve was linear from 1.2x10-4 to 1.0x10-3 mol L-1. The evaluation of biosensor lifetime leads to 500 injections. Commercial pharmaceutical samples were analyzed with the proposed method and the results were compared with those obtained by high-performance liquid chromatography (HPLC).

Leaching of Picloram in ultisol under different rainfall volumes

The present study aimed to evaluate the leaching potential of Picloram in Ultisol columns under different rainfall amounts. For such, 30 treatments were evaluated (one soil associated with three levels of rainfall and ten depths).The experiments were arranged in a split-plot design, in a completely randomized design, with four replications. PVC columns of 10 cm in diameter and 50 cm in length were filled with these soils, moistened, and placed upright for 48 hours to drain the excess water. The herbicide was applied and rainfall simulations were carried out at specified intensities, according to the treatments, to force Picloram leaching. After 72 hours, all the columns were arranged in a horizontal position and opened lengthwise. Then, soil sampling was carried out every 5 cm of depth for subsequent herbicide extraction and quantification and analysis by high performance liquid chromatography. The remaining soil samples were placed in plastic pots, and, at the respective depths, the indicator species Cucumis sativus was sown. Twenty-one days after the emergence (DAE) of the indicator plants, evaluations were conducted to verify the symptoms of toxicity caused by Picloram in the plants. It was concluded that Picloram leaching is directly dependent on the volume of rain applied. The herbicide reached the deepest regions in the soil with the highest intensity of rain. The results obtained by bioassay were in agreement with those found by liquid chromatography.

Allelopathic effect of black mustard tissues and root exudates on some crops and weeds

Laboratory and greenhouse experiments were conducted to evaluate the phytotoxic effect of black mustard extracts and root exudates on two crops: Trifolium alexandrinum and Triticum aestivum, and two weeds: Phalaris paradoxa and Sisymbrium irio. The seeds were treated with aqueous and ethanolic extracts and chloroform for eight days, or subjected to root exudates of just harvested mustard in a greenhouse for five weeks. High-performance liquid chromatography (HPLC) was used to quantify phytotoxins from plant tissues. Seed germination of P. paradoxa was reduced with the lowest concentration of the different extracts. However, the aqueous extract at 4% completely curtailed the germination of all the target species. In general, plant extracts had a concentration-dependent reduction of seedling growth of the target species. However, the ethanolic extract, at the lowest concentration, has stimulated the shoot length of both T. alexandrinum and T. aestivum, and the root length of the former. Mustard root exudates inhibited emergence and growth of the target species throughout the experiment. Ferulic and syringic acids were the dominant allelochemicals found when HPLC was used.

Influence of spray mixture volume and flight height on herbicide deposition in aerial applications on pastures

The objective of the present study was to analyze the influence of spray mixture volume and flight height on herbicide deposition in aerial applications on pastures. The experimental plots were arranged in a pasture area in the district of Porto Esperidião (Mato Grosso, Brazil). In all of the treatments, the applications contained the herbicides aminopyralid and fluroxypyr (Dominum) at the dose of 2.5 L c.p. ha-1, including the adjuvant mineral oil (Joint Oil) at the dose of 1.0 L and a tracer to determine the deposition by high-performance liquid chromatography (HPLC) (rhodamine at a concentration of 0.6%). The experiment consisted of nine treatments that comprised the combinations of three spray volumes (20, 30 and 50 L ha-1) and three flight heights (10, 30 and 40 m). The results showed that, on average, there was a tendency for larger deposits for the smallest flight heights, with a significant difference between the heights of 10 and 40 m. There was no significant difference among the deposits obtained with the different spray mixture volumes.

Shikimate accumulation, glyphosate absorption and translocation in horseweed biotypes

In Brazil, few research works on mechanisms of weed resistance to glyphosate have been conducted so far. Therefore, this research aimed to study analytical procedures determining the relation between the concentration of plant shikimate after glyphosate application and the plant resistance to this herbicide; and evaluate the glyphosate absorption and translocation into two resistant ® and susceptible (S) horseweed biotypes to glyphosate. Horseweed plants with nine true leaves received glyphosate (720 g a.e. ha-1), and 2, 3, 4, 7 and 10 days after application (DAA) the concentration of shikimic acid was measured by HPLC. In another experiment, plants were treated with radiolabeled glyphosate (14C) (1.456 MBq mmol-1 specific activity) and radioactivity was measured 4, 8, 24, 48 and 72 hours after treatment (HAT) by liquid scintillation spectrometry. The shikimate concentration in plants increased 16,351.14 and 7,892.25 mg kg-1 of dry weight, for R and S plants respectively, at seven DAA. Therefore, the procedure for quantification of shikimic acid was suitable for R and S plants differentiation to glyphosate, indicating that the R population is actually resistant to glyphosate. On average, 98% of glyphosate applied was absorbed by the studied biotypes, at 72 HAT. Around 68% of the absorbed radioactivity remained on the biotypes leaves treated, the S biotype showing the highest translocation. Therefore, the R biotype resistance mechanism studied is associated to the differential translocation.

Leaching of Sulfentrazone in Soils from the Sugarcane Region in the Northeast Region of Brazil

Sulfentrazone leaching potential is dependent on soil properties such as strength and type of clay, organic matter content and pH, and may result in ineffectiveness of the product and contamination of groundwater. The objective of this study was to evaluate sulfentrazone leaching in five soils of the sugarcane region in the Northeast Region of Brazil, with different physical and chemical properties, by means of bioassay and high-performance liquid chromatography (HPLC) resolution. The experiment was conducted in a split plot in a completely randomized design. The plots had PVC columns with a 10 cm diameter and being 50 cm deep, filled with five different soil classes (quartzarenic neosol, haplic cambisol, yellowish-red latosol, yellowish-red acrisol, and haplic gleysol), and subplots for 10 depths in columns, 5 cm intervals. On top of the columns, sulfentrazone application was conducted and 12 hours later there was a simulated rainfall of 60 mm. After 72 hours, the columns were horizontally placed and longitudinally open, divided into sections of 5.0 cm. In the center of each section of the columns, soil samples were collected for chromatographic analyses and sorghum sowing was carried out as an indicator plant. The bioassay method was more sensitive to detect the presence of sulfentrazone in an assessment for chromatography soil, having provided greater herbicide mobility in quartzarenic neosol and yellowish-red latosol, whose presence was detected by the indicator plant to a depth of 45 and 35 cm, respectively. In the other soils, sulfentrazone was detected up to 20 cm deep. The intense mobility of sulfentrazone in quartzarenic neosol may result in herbicide efficiency loss in the soil because the symptoms of intoxication and the amount of herbicide detected via silica were highest between 15 cm and 35 cm depth regarding the soil surface layer (0-10 cm), indicating that sulfentrazone should be avoided in soils with such characteristics.