Analytica Chimica Acta (v.609, #1)

Contents (v-vi).

A Diltiazem kinetic spectrophotometric method was optimized by factorial analysis. The experimental method is based on a two-stage reaction of Diltiazem with hydroxylamine and a ferric salt: in the first stage there is a hydroxamic acid formation; and, in the second stage there is a red colour complex ferric hydroxamate formation. The variables under investigation were: solvent; hydroxylamine, sodium hydroxide and ammonium ferric sulphate concentrations; volume of perchloric acid; and, temperature. The responses of the reactional system were the maximum absorbance, the wavelength and the reaction time at maximum absorbance. Experimental design methodologies were used in the optimization. Fractional and full factorial designs followed by optimization Box-Behnken and central composite experimental designs were used. The observed optimum conditions were: methanol as reaction solvent; hydroxylamine concentration of 9.375%; sodium hydroxide concentration of 18.750%; ferric reagent concentration of 2.000%; minimum volume of perchloric acid to neutralize the sodium hydroxide; and, room temperature as reaction temperature. With this set of experimental conditions a reaction time of 10.5 s with maximum colour development at 512 nm wavelength was achieved.
Keywords: Antihypertensor Diltiazem; UV–Vis spectroscopy; Kinetic method; Optimization; Experimental design methodologies;

The use of some unconventional non-linear modeling techniques, i.e. classification and regression trees and multivariate adaptive regression splines-based methods, was explored to model the blood–brain barrier (BBB) passage of drugs and drug-like molecules. The data set contains BBB passage values for 299 structural and pharmacological diverse drugs, originating from a structured knowledge-based database. Models were built using boosted regression trees (BRT) and multivariate adaptive regression splines (MARS), as well as their respective combinations with stepwise multiple linear regression (MLR) and partial least squares (PLS) regression in two-step approaches. The best models were obtained using combinations of MARS with either stepwise MLR or PLS. It could be concluded that the use of combinations of a linear with a non-linear modeling technique results in some improved properties compared to the individual linear and non-linear models and that, when the use of such a combination is appropriate, combinations using MARS as non-linear technique should be preferred over those with BRT, due to some serious drawbacks of the BRT approaches.
Keywords: Quantitative structure–activity relationships; Blood–brain barrier passage; In silico prediction; Boosted regression trees; Multivariate adaptive regression splines; Two-step approaches;

Support vector machines (SVM), radial basis function neural networks (RBFNN) and multiple linear regression (MLR) methods were used to investigate the correlation between GC retention indexes (RI) and physicochemical descriptors for both 174 and 132 diverse organic compounds. The correlation coefficient r 2 between experimental and predicted retention index for training and test sets by SVM, RBFNN and MLR is 0.986, 0.976 and 0.971 (for 174 compounds), 0.986, 0.951 and 0.963 (for 132 compounds) respectively. The results show that non-linear SVM derives statistical models have similar prediction ability to those of RBFNN and MLR methods. This indicates that SVM can be used as an alternative modeling tool for quantitative structure–property/activity relationship (QSPR/QSAR) studies.
Keywords: Support vector machines; Radial basis neural networks; Multiple linear regression; Gas chromatography retention index;

The application of conducting polymer nanoparticle electrodes to the sensing of ascorbic acid by Adriano Ambrosi; Aoife Morrin; Malcolm R. Smyth; Anthony J. Killard (37-43).
An ascorbic acid sensor was fabricated via the drop-casting of dodecylbenzene sulphonic acid (DBSA)-doped polyaniline nanoparticles onto a screen-printed carbon-paste electrode. The modified electrode was characterised with respect to the numbers of drop cast layers, optimum potential and operating pH. The sensor was found to be optimal at neutral pH and at 0 V vs. Ag/AgCl. Under these conditions, the sensor showed good selectivity and sensitivity in that it did not respond to a range of common interferents such as dopamine, acetaminophen, uric acid and citric acid, but was capable of the detection of ascorbic acid at a sensitivity of 0.76 μA mM−1 or 10.75 μA mM−1  cm−2 across a range from 0.5 to 8 mM (r 2  = 0.996, n  = 6), and a limit of detection of 8.3 μM (S/N = 3). The sensor was compared to a range of other conducting polymer-based ascorbate sensors and found to be comparable or superior in terms of analytical performance.
Keywords: Polyaniline; Conducting polymer; Dodecylbenzene sulphonic acid; Nanoparticle; Ascorbic acid; Sensor;

We have developed a multiwalled carbon nanotube/dihydropyran (MWCNT/DHP) composite sensor for the electrochemical detection of insulin in a microfluidic device. This sensor has been employed for physiological measurements of secreted insulin from pancreatic islets in a Cytosensor® previously modified to be a multianalyte microphysiometer (MAMP). When compared with other established electrochemical insulin sensors, the MWCNT/DHP composite film sensor presented improved resistance to fluidic shear forces, while achieving enhanced electrode kinetics. In addition, the preparation of the composite film is straightforward and facile with a self-polymerizing monomer, DHP, used to add mechanical stability to the film. The sensor film was able to detect insulin concentrations as low as 1 μM in the MAMP during calibration experiments. The MWCNT/DHP composite sensor has been successfully used for the direct detection of insulin secreted by islets in the microphysiometer.
Keywords: Insulin; Multiwalled carbon nanotube; Multianalyte microphysiometer; Cytosensor®; Dihydropyran; Sensor;

A novel method, dispersive liquid–liquid microextraction (DLLME) coupled with high-performance liquid chromatography-variable wavelength detector (HPLC-VWD), has been developed for the determination of three phthalate esters (dimethyl phthalate (DMP), diethyl phthalate (DEP), and di-n-butyl phthalate (DnBP)) in water samples. A mixture of extraction solvent (41 μL carbon tetrachloride) and dispersive solvent (0.75 mL acetonitrile) were rapidly injected into 5.0 mL aqueous sample for the formation of cloudy solution, the analytes in the sample were extracted into the fine droplets of CCl4. After extraction, phase separation was performed by centrifugation and the enriched analytes in the sedimented phase were determined by HPLC-VWD. Some important parameters, such as the kind and volume of extraction solvent and dispersive solvent, extraction time and salt effect were investigated and optimized. Under the optimum extraction condition, the method yields a linear calibration curve in the concentration range from 5 to 5000 ng mL−1 for target analytes. The enrichment factors for DMP, DEP and DnBP were 45, 92 and 196, respectively, and the limits of detection were 1.8, 0.88 and 0.64 ng mL−1, respectively. The relative standard deviations (R.S.D.) for the extraction of 10 ng mL−1 of phthalate esters were in the range of 4.3–5.9% (n  = 7). Lake water, tap water and bottled mineral water samples were successfully analyzed using the proposed method.
Keywords: Dispersive liquid–liquid microextraction; High-performance liquid chromatography; Phthalate esters; Water samples;

This paper describes micropatterning of proteins on the surface of three-dimensional hydrogel microstructures. Poly(ethylene glycol) (PEG)-based hydrogel microstructures were fabricated on a glass substrate using a poly(dimethylsiloxane) (PDMS) replica as a molding insert and photolithography. The lateral dimension and height of the hydrogel microstructures were easily controlled by the feature size of the photomask and depth of the PDMS replica, respectively. Bovine serum albumin (BSA), a model protein, was covalently immobilized to the surface of the hydrogel microstructure via a 5-azidonitrobenzoyloxy N-hydroxysuccinimide bifunctional linker at a surface density of 1.48 mg cm−2. The immobilization of BSA on the PEG hydrogel surface was demonstrated with XPS by confirming the formation of a new nitrogen peak, and the selective immobilization of fluorescent-labeled BSA on the outer region of the three-dimensional hydrogel micropattern was demonstrated by fluorescence. A hydrogel microstructure could immobilize two different enzymes separately, and sequential bienzymatic reaction was demonstrated by reacting glucose and Amplex Red with a hydrogel microstructure where glucose oxidase was immobilized on the surface and peroxidase was encapsulated. Activity of immobilized glucose oxidase was 16.5 U mg−1, and different glucose concentration ranged from 0.1 to 20 mM could be successfully detected.
Keywords: Hydrogel microstructure; Micropatterning; Protein immobilization; Bienzymatic reaction;

Detection of carbohydrates using new labeling reagent 1-(2-naphthyl)-3-methyl-5-pyrazolone by capillary zone electrophoresis with absorbance (UV) by Jinmao You; Xiao Sheng; Chenxu Ding; Zhiwei Sun; Yourui Suo; Honglun Wang; Yulin Li (66-75).
A novel labeling reagent 1-(2-naphthyl)-3-methyl-5-pyrazolone (NMP) coupled with capillary electrophoresis (CE) with DAD detection for the determination of carbohydrates has been developed. The chromophore in the 1-phenyl-3-methyl-5-pyrazolone (PMP) reagent is replaced by naphthyl functional group, which results in a reagent with very high molar absorptivity (ɛ 251 nm  = 5.58 × 104  L mol−1  cm−1). This permits NMP-labeled carbohydrates to be detected with UV absorbance in standard 50-μm-i.d. fused silica capillaries by zone electrophoresis. In this mode, nanomolar concentrations of detection limits are obtained. The method for the derivatization of carbohydrates with NMP is simplified. The derivatization reaction is rapid and mild in the presence of ammonia catalyst without further transfer steps. Nine monosaccharide derivatives such as mannose, galacturonic acid, glucuronic acid, rhamnose, glucose, galactose, xylose, arabinose and fucose can successfully be detected in CE mode. Good reproducibility can be obtained with relative standard deviation (R.S.D.) values of the migration times and peak area, respectively, from 0.44 to 0.48 and from 3.2 to 4.8. Furthermore, the developed method has been successfully applied to the analysis of carbohydrates in the hydrolyzed rape bee pollen samples.
Keywords: Capillary electrophoresis (CE); Carbohydrates; 1-(2-Naphthyl)-3-methyl-5-pyrazolone (NMP); Derivatization;

Open-tubular gas chromatography using capillary coated with octadecylamine-capped gold nanoparticles by Qi-Shu Qu; Fei Shen; Ming Shen; Xiao-Ya Hu; Gong-Jun Yang; Cheng-Yin Wang; Chao Yan; Yu-Kui Zhang (76-81).
The octadecylamine-capped gold nanoparticles (ODA-Au-NPs) were prepared and directly used to coat the capillary wall. The hydrophobic coating acted as the stationary phase for open-tubular gas chromatography (OTGC). The ODA-Au-NPs can be adsorbed tightly onto the inner surface of fused silica capillary column via electrostatic interaction and enhanced interaction of van der Waals between gold nanoparticles and the capillary wall. Thus, the modification of the inner surface of capillary column by ODA-Au-NPs can be achieved simply by flushing the capillary with a solution of ODA-Au-NPs and the resulted ODA-Au-NPs coating is very stable. No perceptible degradation in the ODA-Au-NPs-based separation was observed after ∼1900 sample runs. This type of columns also provided excellent chromatographic performances: high number of theoretical plates, outstanding run-to-run and column-to-column reproducibility, and high selectivity for a wide range of test mixtures. An efficiency of 2474 theoretical plates per meter for chlorobenzene was obtained on an ODA-Au-NPs-modified 1.6 m × 100 μm i.d. fused silica capillary column.
Keywords: Octadecylamine; Gold nanoparticle; Open-tubular gas chromatography;

In the present study, a simple and sensitive methodology based on dynamic headspace solid-phase microextraction (HS-SPME) followed by thermal desorption gas chromatography with quadrupole mass detection (GC–qMSD), was developed and optimized for the determination of volatile (VOCs) and semi-volatile (SVOCs) compounds from different alcoholic beverages: wine, beer and whisky. Key experimental factors influencing the equilibrium of the VOCs and SVOCs between the sample and the SPME fibre, as the type of fibre coating, extraction time and temperature, sample stirring and ionic strength, were optimized. The performance of five commercially available SPME fibres was evaluated and compared, namely polydimethylsiloxane (PDMS, 100 μm); polyacrylate (PA, 85 μm); polydimethylsiloxane/divinylbenzene (PDMS/DVB, 65 μm); carboxen™/polydimethylsiloxane (CAR/PDMS, 75 μm) and the divinylbenzene/carboxen on polydimethylsiloxane (DVB/CAR/PDMS, 50/30 μm) (StableFlex).An objective comparison among different alcoholic beverages has been established in terms of qualitative and semi-quantitative differences on volatile and semi-volatile compounds. These compounds belong to several chemical families, including higher alcohols, ethyl esters, fatty acids, higher alcohol acetates, isoamyl esters, carbonyl compounds, furanic compounds, terpenoids, C13-norisoprenoids and volatile phenols. The optimized extraction conditions and GC–qMSD, lead to the successful identification of 44 compounds in white wines, 64 in beers and 104 in whiskys. Some of these compounds were found in all of the examined beverage samples.The main components of the HS-SPME found in white wines were ethyl octanoate (46.9%), ethyl decanoate (30.3%), ethyl 9-decenoate (10.7%), ethyl hexanoate (3.1%), and isoamyl octanoate (2.7%). As for beers, the major compounds were isoamyl alcohol (11.5%), ethyl octanoate (9.1%), isoamyl acetate (8.2%), 2-ethyl-1-hexanol (5.9%), and octanoic acid (5.5%). Ethyl decanoate (58.0%), ethyl octanoate (15.1%), ethyl dodecanoate (13.9%) followed by 3-methyl-1-butanol (1.8%) and isoamyl acetate (1.4%) were found to be the major VOCs in whisky samples.
Keywords: Volatile and semi-volatile compounds; Headspace solid-phase microextraction; White wine; Beer; Whisky; GC–qMSD;

Magnetic particles coated with specific biomolecules are often used as solid supports for bioassays but conventional test tube based techniques are time consuming and labour intensive. An alternative is to work on magnetic particle plugs immobilised inside microfluidic channels. Most research so far has focussed on immobilising one type of particle to perform one type of assay. Here we demonstrate how several assays can be performed simultaneously by flushing a sample solution over several plugs of magnetic particles with different surface coatings. Within a microchannel, three plugs of magnetic particles were immobilised with external magnets. The particles featured surface coatings of glycine, streptavidin and protein A, respectively. Reagents were then flushed through the three plugs. Molecular binding occurred between matching antigens and antibodies in continuous flow and was detected by fluorescence. This first demonstration opens the door to a quicker and easier technique for simultaneous bioassays using magnetic particles.
Keywords: Magnetic particles; Microfluidics; Assay; Simultaneous analysis;

Steady-state diffusion regime in solid-phase microextraction kinetics by Karim Benhabib; Thomas L. ter Laak; Herman P. van Leeuwen (113-119).
The temporal evolution of diffusion-controlled analyte accumulation in solid-phase microextraction (SPME) is critically discussed in terms of the various aspects of steady-state diffusion in the two phases under conditions of fast exchange of the analyte at the solid phase film/water interface. For partition coefficients (K sw) much larger than unity and a sufficiently thin polymer film, the concentration gradient of the analyte in the polymer phase is largely insignificant. The growth of the accumulated amount of analyte is then adequately described by the well-known exponential expression for steady-state diffusion under non-depletive conditions, provided the initial transient stage is properly taken into account. In case of fiber-type solid phases, the cylindrical nature of the diffusion complicates the nature of the transient stage as well as the magnitude of the steady-state flux.
Keywords: Solid-phase microextraction; Atrazine; Kinetics; Steady-state; Analytical model;

Use of polystyrene spin-coated compact discs for microimmunoassaying by Jesús Tamarit-López; Sergi Morais; Rosa Puchades; Ángel Maquieira (120-130).
The analytical potential of polystyrene (PS) spin-coated modified compact discs (CDs) surface as platforms for the development of microarray immunoassays is presented. The surface maintained the optical characteristics of compact discs, obtaining a transparent and smooth film polymer of 70 nm thickness, the track being read (λ 780 nm) without errors in a commercial CD reader/writer. The analytical capability of the methodology was demonstrated through an analysis of a neurotoxic compound (2560 spots per disc), reaching 0.08 μg L−1 as limit of detection. These figures demonstrate the enormous potential of using PS spin-coated compact discs in combination with CD players as an easy-to-operate and portable device to develop lab-on-a-disc analytical applications.
Keywords: Compact disc; Polystyrene; Microimmunoassay; Chlorpyriphos;