Development of a fast method for simultaneous determination of hippuric acid, mandelic acid, and creatinine in urine by capillary zone electrophoresis using polymer multilayer-coated capillary.

This paper reports the development of a fast separation method employing capillary zone electrophoresis for the simultaneous determination of hippuric acid, mandelic acid, and creatinine in samples of urine using a coated capillary. The background electrolyte was composed of 10 mmol L-1 tris(hydroxymethyl)aminomethane and 30 mmol L-1 2-hydroxy-isobutyric acid at pH 3.6. The internal standard was 3,5-dihydroxybenzoic acid. Separations were performed in a fused silica capillary (32 cm total length, 8.5 cm effective length, and 50 µm internal diameter) coated with crosslinked hydroxypropyltrimethyl ammonium chloride chitosan and κ-carrageenan. Direct UV detection was performed at a wavelength of 200 nm. Samples and standards were injected hydrodynamically (-50 mbar, 3 s) using the short-end injection procedure. The electrophoretic system was operated under constant voltage of 30 kV with positive polarity on the injection side. The separation time for hippuric acid, mandelic acid, and creatinine was less than 70 s. The evaluation of some analytical parameters of the method for the three analytes showed good linearity (R 2 > 0.99), limit of detections of 0.21 to 0.63 mg L-1, inter-day precision better than 3.0% (peak area), and recovery in the range of 98 to 106%. The method developed was applied in the analysis of the three analytes in urine samples. Graphical Abstract New method using capillary zone electrophoresis for analysis of creatinine, hippuric acid and mandelic acid in urine.

New multilayer coating using quaternary ammonium chitosan and κ-carrageenan in capillary electrophoresis: application in fast analysis of betaine and methionine.

The aim of this study was to develop a new multilayer coating with crosslinked quaternary ammonium chitosan (hydroxypropyltrimethyl ammonium chloride chitosan; HACC) and κ-carrageenan for use in capillary electrophoresis. A new semi-permanent multilayer coating was formed using the procedure developed and the method does not require the presence of polymers in the background electrolyte (BGE). The new capillary multilayer coating showed a cathodic electroosmotic flow (EOF) of around 30×10(-9) m(2) V(-1) s(-1) which is pH-independent in the range of pH 2 to 10. The enhanced EOF at low pH obtained contributed significantly to the development of a fast method of separation. The multilayer coating was then applied in the development of a fast separation method to determine betaine and methionine in pharmaceutical formulations by capillary zone electrophoresis (CZE). The BGE used to determine the betaine and methionine concentrations was composed of 10 mmol L(-1) tris(hydroxymethyl) aminomethane, 40 mmol L(-1) phosphoric acid and 10% (v/v) ethanol, at pH 2.1. A fused-silica capillary of 32 cm (50 µm ID×375 µm OD) was used in the experiments and samples and standards were analyzed employing the short-end injection procedure (8.5 cm effective length). The instrumental analysis time of the optimized method was 1.53 min (approx. 39 runs per hour). The validation of the proposed method for the determination of betaine and methionine showed good linearity (R(2)>0.999), adequate limit of detection (LOD <8 mg L(-1)) for the concentration in the samples and inter-day precision values lower than 3.5% (peak area and time migration). The results for the quantification of the amino acids in the samples determined by the CZE-UV method developed were statistically equal to those obtained with the comparative LC-MS/MS method according to the paired t-test with a confidence level of 95%.

Using multiple short-end injections to develop fast electrophoretic separations--applications in iodide analysis.

The aim of this study was to develop a fast CE separation method by using multiple short-end injections in a capillary coated with quaternary ammonium chitosan (HACC), in order to determine the iodide content of pharmaceutical formulations. The BGE was composed of 20 mM tris(hydroxymethyl)aminomethane and 11 mM hydrochloric acid, at pH 8. The internal standard used was thiocyanate. Separations were performed in a fused silica capillary (32 cm total length, 8.5 cm effective length and 50 µm i.d.) coated with HACC and direct UV detection at 220 nm. EOF was modified by flushing the capillary with polymeric solution, resulting in a semi-permanent coating of controlled and stable EOF. The EOF was anodic at pH 8. Different strategies, using single and multiple injection short-end configurations, were studied to develop a CE method that resulted in a maximum number of iodide samples analyzed per hour: one plug and flush (Sflush) 35 samples/h, one plug without flush (SWflush) 76 samples/h, four plugs and flush (Mflush) 61 samples/h, and four plugs without flush (MWflush) 80 samples/h. Using the multiple injection configuration, it was possible to inject up to four plugs using spacer electrolytes with good separation efficiency and selectivity. The voltage application time needed to separate the eight peaks (iodide and thiocyanate) with MWflush was only 12s. The method was validated and samples were analyzed using MWflush. Good linearity (R(2)>0.999); a limit of detection 0.4 mg L(-1); intermediate precision better than 3.8% (peak area) and recovery in the range of 99-102% were obtained.

A new method to determine biological sample volume by short end multiple injection capillary electrophoresis: application in determination of nitrate and thiocyanate in human saliva.

The aim of this study was to develop a simple and rapid method of capillary electrophoresis using a short end multiple injection in free solution to determine simultaneously the biological sample volume and analytes concentration. The method consists of a sequence of injection steps with an internal standard as the reference for correction of the volume of sample collected. The procedure was applies in the determination of NO(3)(-) and SCN in saliva samples. The background electrolyte was composed of 12 mM tris(hydroxymethyl)aminomethane and 8.5mM sulfuric acid, at pH 2.5. The internal standard used was BrO(3)(-). A fused silica capillary (48.5 cm total length, 8.5 cm effective length and 75 µm i.d.) coated with chitosan was used in a short-end injection configuration. Modification of the electroosmotic flow (EOF) using dynamic coating resulted in a controlled and stable EOF, contributing to the rapid separation of anions (0.36 min) in co-electroosmotic mode. The validation of the method for correcting the volume of saliva collected with a swab showed a difference of less than 3.5% compared with the predicted value and a correlation of 0.999. The limits of detection for NO(3)(-) and SCN(-) were 0.13 and 0.23 mg L(-1), respectively. The inter-day precision of the method determined for both analytes was less than 5% and the recovery ranged between 97 and 102%.

Adsorption and desorption of Cu(II), Cd(II) and Pb(II) ions using chitosan crosslinked with epichlorohydrin-triphosphate as the adsorbent.

In this study, chitosan (CTS) was crosslinked with both epichlorohydrin (ECH) and triphosphate (TPP), by covalent and ionic crosslinking, respectively. The resulting new CTS-ECH-TPP adsorbent was characterized by CHN analysis, EDS, FTIR spectroscopy, TGA and DSC, and the adsorption and desorption of Cu(II), Cd(II) and Pb(II) ions in aqueous solution were investigated. Potentiometric studies were also performed and revealed three titratable protons for each pK(a) value of 5.14, 6.76 and 9.08. The results obtained showed that the optimum pH values for adsorption were 6.0 for Cu(II), 7.0 for Cd(II) and 5.0 for Pb(II). The kinetics study demonstrated that the adsorption process proceeded according to the pseudo-second-order model. Three isotherm models (Langmuir, Freundlich and Dubinin-Radushkevich) were employed in the analysis of the adsorption equilibrium data. The Langmuir model resulted in the best fit and the new adsorbent had maximum adsorption capacities for Cu(II), Cd(II) and Pb(II) ions of 130.72, 83.75 and 166.94 mg g(-1), respectively. Desorption studies revealed that HNO(3) and HCl were the best eluents for desorption of Cu(II), Cd(II) and Pb(II) ions from the crosslinked chitosan.

Evaluation of the potential of chitosan hydrogels to extract polar organic species from nonpolar organic solvents: application to the extraction of aminopyridines from hexane.

The extraction of a series of aminopyridines (APs) utilizing chitosan hydrogels in hexane was investigated. The chitosan hydrogel was prepared using glutaraldehyde as a cross-linking agent. Experiments were carried out to determine the maximum extraction efficiency, distribution coefficient, sorption capacity, and adsorption and desorption mechanisms. The efficiency of extraction of aminopyridines attained a maximum value of ca. 100% with the distribution coefficients for the transfer of the aminopyridines from hexane to chitosan hydrogel increasing in the order of ortho-

Textile effluents induce biomarkers of acute toxicity, oxidative stress, and genotoxicity.

The present work consists of a comparative evaluation of the toxicity of a nonremediated textile effluent (NRTE) with an effluent remediated by a pulverized chitosan system (RCTS) or by a conventional effluent process (remediated biologic and physico-chemical effluent [RBPC]). Acute toxicity assays, oxidative stress biomarkers, physico-chemical parameters, and genotoxicity indices were analyzed to achieve the toxicity of all effluents. After RCTS treatment, approximately 80% of dyes were removed, together with a significant decreased of the metal content, compared with a relatively increase in metal content after RBPC treatment. RBPC and RCTS treatments did not cause acute toxicity to Vibrio fischeri and Artemia sp., whereas RBPC caused acute toxicity to Daphnia magna but RCTS did not. Compared with NRTE, chitosan remediation decreased oxidative stress biomarkers, such as the contents of lipoperoxidation (measured as thiobarbituric acid-reactive substances [TBARS], 29.9%) and the reduced form of glutathione (GSH; 73.5%) levels in D. rerio, whereas animals exposed to RBPC showed enhanced TBARS (57.2%) and decreased GSH concentrations (56.4%). RCTS and RBPC remediation elicited catalase activity induction (161.8% and 127.3%, respectively) compared with NRTE. Accordingly, DNA fragmentation and micronucleus frequency in D. rerio decreased after remediation with RBPC or RCTS compared with NRTE, but RCTS treatment was more effective than RBPC in decreasing genotoxicity (90.5% and 73.8% decrease in DNA fragmentation and 67.8% and 50.4% decrease in micronucleus frequency, respectively). The results indicate that chitosan adsorption system is a useful tool for textile effluent remediation compared with the conventional remediation by biologic and physico-chemical processes.

Cross-linked quaternary chitosan as an adsorbent for the removal of the reactive dye from aqueous solutions.

Adsorption of reactive orange 16 by quaternary chitosan salt (QCS) was used as a model to demonstrate the removal of reactive dyes from textile effluents. The polymer was characterized by infrared (IR), energy dispersive X-ray spectrometry (EDXS) analyses and amount of quaternary ammonium groups. The adsorption experiments were conducted at different pH values and initial dye concentrations. Adsorption was shown to be independent of solution pH. Three kinetic adsorption models were tested: pseudo-first-order, pseudo-second-order and intraparticle diffusion. The experimental data best fitted the pseudo-second-order model, which provided a constant velocity, k2, of 9.18 x 10(-4)g mg(-1)min(-1) for a 500 mg L(-1) solution and a value of k2, of 2.70 x 10(-5)g mg(-1)min(-1) for a 1000 mg L(-1) solution. The adsorption rate was dependent on dye concentration at the surface of the adsorbent for each time period and on the amount of dye adsorbed. The Langmuir isotherm model provided the best fit to the equilibrium data in the concentration range investigated and from the isotherm linear equation, the maximum adsorption capacity determined was 1060 mg of reactive dye per gram of adsorbent, corresponding to 75% occupation of the adsorption sites. The results obtained demonstrate that the adsorbent material could be utilized to remove dyes from textile effluents independent of the pH of the aqueous medium.

Spray-dried chitosan microspheres containing 8-hydroxyquinoline -5 sulphonic acid as a new adsorbent for Cd(II) and Zn(II) ions.

In the present study, a new chelating adsorbent was prepared from chitosan microspheres cross-linked with glutaraldehyde by spray drying using 8-hydroxyquinoline -5 sulphonic acid as chelant agent (CTS-SX-CL). Microspheres of the new adsorbent were characterized by Raman spectroscopy, scanning electron microscopy (SEM) and energy-dispersive X-ray microanalysis (EDX). The effect of pH, contact time and concentration of metallic ions in solution were evaluated on the adsorption behavior of Cd(II) and Zn(II) by CTS-SX-CL. Adsorption was maximum for both Cd(II) and Zn(II) at pH 8.0. Adsorption kinetic curves were obtained and could be fit by the pseudo second-order adsorption model. An analysis of equilibrium adsorption data using the Langmuir isotherm model indicated that the maximum adsorption capacity of CTS-SX-CL was higher than that of CTS-CL for both ions investigated. The adsorption capacity increased 74% for Cd(II).

Reduction of acidity and removal of metal ions from coal mining effluents using chitosan microspheres.

Effluents from coal mining operations are not only highly acid but also depict elevated concentrations of metals which may contaminate the environment. Due to the polybasic characteristic of chitosan, this biopolymer is capable of both neutralizing and removing iron, aluminum and copper ions from such effluents. The present study aimed at evaluating the use of chitosan microspheres for their importance in continuous systems. The microspheres were prepared by the phase inversion method. Their average diameter and morphology were determined. Water samples from decantation pool (DP) and acidic mine drainage (AMD) effluents were treated using different amounts of microspheres. The pH and concentration of Fe, Al and Cu ions were evaluated both before and after treatment of effluent samples. The results revealed that the microspheres were capable of increasing the pH of DP and AMD samples from 2.34 and 2.58, respectively, to 6.20, i.e., close to neutrality. The treatment also resulted in full removal of the metals investigated.

Kinetics and equilibrium adsorption of Cu(II), Cd(II), and Ni(II) ions by chitosan functionalized with 2[-bis-(pyridylmethyl)aminomethyl]-4-methyl-6-formylphenol.

Chitosan biopolymer chemically modified with the complexation agent 2[-bis-(pyridylmethyl)aminomethyl]-4-methyl-6-formylphenol (BPMAMF) was employed to study the kinetics and the equilibrium adsorption of Cu(II), Cd(II), and Ni(II) metal ions as functions of the pH solution. The maximum adsorption of Cu(II) was found at pH 6.0, while the Cd(II) and Ni(II) maximum adsorption occurred in acidic media, at pH 2.0 and 3.0, respectively. The kinetics was evaluated utilizing the pseudo-first-order and pseudo-second-order equation models and the equilibrium data were analyzed by Langmuir and Freundlich isotherms models. The adsorption kinetics follows the mechanism of the pseudo-second-order equation for all studied systems and this mechanism suggests that the adsorption rate of metal ions by CHS-BPMAMF depends on the number of ions on the adsorbent surface, as well as on their number at equilibrium. The best interpretation for the equilibrium data was given by the Langmuir isotherm and the maximum adsorption capacities were 109 mg g-1 for Cu(II), 38.5 mg g-1 for Cd(II), and 9.6 mg g-1 for Ni(II). The obtained results show that chitosan modified with BPMAMF ligand presented higher adsorption capacity for Cu(II) in all studied pH ranges.

Sulphoxine immobilized onto chitosan microspheres by spray drying: application for metal ions preconcentration by flow injection analysis.

A new chelating resin based on chitosan biopolymer modified with 5-sulphonic acid 8-hydroxyquinoline using the spray drying technique for immobilization is proposed. The chelating resin was characterized by thermogravimetric analysis (TGA) and X-ray diffraction (XRD) and surface area by nitrogen sorption. The efficiency of the chelating resin was evaluated by the preconcentration of metal ions Cu(II) and Cd(II) present in aqueous samples in trace amounts. The metal ions were previously enriched in a minicolumn and the concentrations of the analytes were determined on-line by flame atomic absorption spectrometry (FAAS). The maximum retention for Cu(II) occurred in the pH range 8-10, and for Cd(II) at pH 7. The optimum flow rate for sorption was found to be 7.2mlmin(-1) for the preconcentration of the metal ions. The analytes gave relative standard deviations (R.S.D.) of 0.7 and 0.6% for solutions containing 20mugl(-1) of Cu(II) and 15mugl(-1) of Cd (II), respectively (n=7). The enrichment factors for Cu(II) and Cd (II) were 19.1 and 13.9, respectively, and the limits of detection (LOD) were 0.2mugl(-1) for Cd(II) and 0.3mugl(-1) for Cu(II), using a preconcentration time of 90s (n=11). The accuracy of the proposed method was evaluated by the metal ion recovery technique, in the analysis of potable water and water from a lake, with recoveries being between 97.2 and 107.3%.

Synthesis of Calcium-Phosphate and Chitosan Bioceramics for Bone Regeneration

Bioceramic composites were obtained from chitosan and hydroxyapatite pastes synthesized at physiological temperature according to two different syntheses approaches. Usual analytical techniques (X-ray diffraction analysis, Fourier transformed infrared spectroscopy, Thermo gravimetric analysis, Scanning electron microscopy, X-ray dispersive energy analysis and Porosimetry) were employed to characterize the resulting material. The aim of this investigation was to study the bioceramic properties of the pastes with non-decaying behavior from chitosan-hydroxyapatite composites. Chitosan, which also forms a water-insoluble gel in the presence of calcium ions, and has been reported to have pharmacologically beneficial effects on osteoconductivity, was added to the solid phase of the hydroxyapatite powder. The properties exhibited by the chitosan-hydroxyapatite composites were characteristic of bioceramics applied as bone substitutes. Hydroxyapatite contents ranging from 85 to 98% (w/w) resulted in suitable bioceramic composites for bone regeneration, since they showed a non-decaying behavior, good mechanical properties and suitable pore sizes