Check out our New Publishers' Select for Free Articles

Analytical and Bioanalytical Chemistry (v.389, #2)

Rewarding and recognising researchers by Sara Shinton (pp. 349-351).

started her professional life as a chemist, following her first degree in chemical and analytical science with a PhD in physical chemistry. After a brief period in postdoctoral research, she discovered an interest in transferable skills whilst teaching undergraduate chemists communication skills. After retraining as a university careers adviser, in 2000 she founded a company ( http://www.shintonconsulting.com ) and now delivers careers guidance and skills training to academic researchers and scientists across the UK and the rest of Europe.

Molecular imprinting by F. Dickert (pp. 353-354).

has held a chair of Analytical Chemistry at the University of Vienna since 1994. His activities are focused on molecular recognition, especially sensitive materials as imprinted polymers for chemical sensors.

Small organic molecular imprinted materials: their preparation and application by Xiaoman Jiang; Na Jiang; Haixia Zhang; Mancang Liu (pp. 355-368).

Molecular imprinting is a technique for preparing polymeric materials that are capable of recognizing and binding the desired molecular target with a high affinity and selectivity. The materials can be applied to a wide range of target molecules, even those for which no natural binder exists or whose antibodies are difficult to raise. The imprinting of small organic molecules (e.g., pharmaceuticals, pesticides, amino acids, steroids, and sugars) is now almost routine. In this review, we pay special attention to the synthesis and application of molecular imprinted polymer (MIPs) imprinted with small organic molecules, including herbicides, pesticides, and drugs. The advantages, applications, and recent developments in small organic molecular imprinted technology are highlighted.

Keywords: Molecular imprinting; Solid-phase extraction; High-performance liquid chromatography; Sensor

Molecularly imprinted beads by surface imprinting by Chau Jin Tan; Yen Wah Tong (pp. 369-376).

Molecular imprinting is a state-of-the-art technique for imparting molecular recognition properties to a synthetic polymeric matrix. Conventionally, the technique is easily carried out using bulk imprinting, where molecularly imprinted polymers (MIPs) are prepared in large chunks and post-treatment processes like grinding and sieving are then required. However, this strategy tends to produce sharp-edged, irregular MIP bits with a limited scope of direct application. In addition, due to the creation of binding sites within the polymeric bulk, the issue of the hindrance of adsorbate diffusion (especially in the case of macromolecules) during template rebinding makes the MIPs prepared through this approach unsuitable for practical applications. Thus over the years, many efforts to address the limitations of conventional molecular imprinting techniques have resulted in new imprinting methodologies. Systems like suspension and precipitation polymerization, where MIPs with tunable morphologies can be prepared, have been developed. Additionally, strategies like surface imprinting have also been employed. Ultimately, both of these approaches have been combined to prepare regularly shaped surface-imprinted MIP beads. Such an approach incorporates the advantages of both methodologies at the same time. Given their desirable physical morphologies and favorable adsorption kinetics, MIPs prepared in this manner show significant promise for industrial applications. Therefore, they will be the main focus of this review.

Keywords: Molecular imprinting; Surface imprinting; Beads; Polymerization; Core–shell; Emulsion

Chiral recognition applications of molecularly imprinted polymers: a critical review by Norbert M. Maier; Wolfgang Lindner (pp. 377-397).

Molecular imprinting technology offers the unique opportunity to tailor chiral stationary phases with predefined chiral recognition properties by employing the enantiomers of interest as binding-site-forming templates. Added advantages, such as ease of preparation, chemical robustness, low-cost production, and the possibility of shaping molecularly imprinted polymers (MIPs) in various self-supporting formats, render them attractive materials for a broad range of chiral recognition applications. In this review a critical overview on recent developments in the field of MIP-based chiral recognition applications is given, focusing on separation techniques and molecular sensing. Inherent limitations associated with the use of enantioselective MIP materials in high-performance separation techniques are outlined, including binding site heterogeneity and slow mass transfer characteristics. The prospects of MIP materials as versatile recognition elements for the design of enantioselective sensor systems are highlighted.

Keywords: Molecular imprinting technology; Chiral recognition; Enantiomer separation; Liquid chromatography; Membrane separation; Batch extraction; Sensors

Molecular imprinting of peptides and proteins in aqueous media by Daniel S. Janiak; Peter Kofinas (pp. 399-404).

Molecular imprinting has received significant attention in recent years, as it provides a viable method for creating synthetic receptors capable of selectively recognizing specific target molecules. Despite significant growth within the field, the majority of template molecules studied thus far have been characterized by their low molecular weight and insolubility in aqueous systems. In biological systems, molecular recognition events occur in aqueous media. Therefore, in order to create molecularly imprinted polymers capable of mimicking biological processes, it is necessary to synthesize artificial receptors which can selectively recognize their respective target biological macromolecules such as peptides and proteins in aqueous media. In this review, we discuss the challenges associated with the imprinting of peptides and proteins in aqueous media. In addition, we discuss the significant progress which has been made within the field.

Keywords: Molecular imprinting; Protein; Peptide; Aqueous; Synthetic receptor

Molecularly imprinted microspheres and nanospheres for di(2-ethylhexyl)phthalate prepared by precipitation polymerization by Jia-Ping Lai; Min-Li Yang; Reinhard Niessner; Dietmar Knopp (pp. 405-412).

Molecularly imprinted microspheres (MIMs, >3 μm) and nanospheres (MINs, ≈450 nm) for the environmental endocrine disruptor di(2-ethylhexyl)phthalate (DEHP) were prepared by a precipitation polymerization (PP) procedure. The effect of the dispersive solvents acetonitrile (ACN) and cyclohexane (CH), the cross-linkers ethylene glycol dimethacrylate (EDMA) and trimethylpropane trimethacrylate (TRIM), and the template on particle size and morphology of polymers was investigated in detail by scanning electron microscopy (SEM) and BET adsorption isotherm determination. When used as HPLC stationary phase, the microspheres exhibited strong affinity for the template DEHP with an imprint factor (IF) higher than 8.0 in ACN/water (60:40, v/v) as mobile phase. Furthermore, baseline separation of DEHP from benzyl butyl phthalate (BBP) and dibutyl phthalate (DBP) could be achieved. In contrast, no or only poor separation could be observed with non-imprinted polymeric polymers (NIPs) or imprinted bulk polymers (MIB), respectively. Similarly, the obtained MINs exhibited an imprinting effect in pure ACN, i.e. the bond amount of DEHP was significantly higher compared to NIPs, as was shown in rebinding experiments. Besides their use as an HPLC stationary phase, MIMs might further be applicable for SPE sample cleanup, while MINs could be used as a recognition layer on sensor surfaces. Figure Molecularly imprinting of di(2-ethylhexyl)phthalate (DEHP)

Keywords: Molecular imprinting; Precipitation polymerization; Microspheres; Nanospheres; Di(2-ethylhexyl)phthalate; HPLC

Molecular recognition of polycyclic aromatic hydrocarbons by pyrene-imprinted microspheres by C. Baggiani; L. Anfossi; P. Baravalle; C. Giovannoli; G. Giraudi (pp. 413-422).

Pyrene-imprinted microbeads that display molecular recognition towards polyaromatic hydrocarbons (PAHs) were obtained by the aqueous suspension thermopolymerization of a mixture of template, 4-vinylpyridine and divinylbenzene in the molar ratio of 1:8:40. The microbeads were packed into an HPLC column and the retention behaviour of pyrene in the presence of eluents of increasing polarity was investigated by measuring the binding capacity and the imprinting factor. Selectivity was evaluated by eluting pyrene and 22 other related PAHs in the HPLC column when equilibrated with acetonitrile–dichloromethane 4:1 (v/v). Twelve molecular descriptors were calculated for each PAH molecule: MW, the molecular weight; SAS, the solvent-accessible molecular surface area; Svdw, the van der Waals molecular surface area; Vol, the van der Waals molecular volume; MOv, the molecular ovality; RG, the radius of gyration; B/L, the breadth-to-length ratio; μ ², the square of the total dipole moment; HOMO, the highest occupied molecular orbital; LUMO, the lowest unoccupied molecular orbital; Δorb, the absolute value of the difference between the HOMO and LUMO; log P, the logarithm of the n-octanol–water partition coefficient. Quantitative structure–retention relationships between the logarithm of the capacity factors and these descriptors were searched for using a multiple linear regression (MLR) method. The best regression models obtained showed that the capacity factor correlated well with those molecular descriptors which had structural character, such as logP, while the effect of the molecular descriptors with electronic character was negligible. The results obtained indicate that the molecular recognition of PAHs by the imprinted polymer is controlled by the shape and dimension of the binding sites through hydrophobic interactions. Figure Retention and imprinting factors of PAHs on the impinted and not-imprinted columns. The area under the lower diagonal line (IF = 1.0) corresponds to the absence of imprinting effect for the interaction between the stationary phase and a given PAH.

Keywords: Molecular imprinting; Molecular recognition; Pyrene; Polycyclic aromatic hydrocarbons; Quantitative structure–retention relationship

Investigating the mechanisms of 17β-estradiol imprinting by computational prediction and spectroscopic analysis by Shuting Wei; Michael Jakusch; Boris Mizaikoff (pp. 423-431).

Molecular dynamics simulations combined with spectroscopic analysis were applied to understand the nature of recognition in molecularly imprinted polymers (MIPs), and for optimizing the MIP formulation. The best monomers for synthesizing imprinted materials for 17β-estradiol (BE2) were selected by evaluating the strength of the template–monomer interaction derived from molecular dynamics simulations. A number of potential functional monomers for BE2 were screened for hydrogen-bonding strength in order to analyze template–monomer interactions favorable for synthesizing noncovalent MIPs, with the simulations revealing that methacrylic acid, 2-(diethylamino)ethyl methacrylate, and methacrylamide provided the highest binding affinity to BE2. These theoretical predictions agree with previously reported results on batch rebinding studies using the corresponding functional monomers for synthesizing a series of MIPs. Molecular analysis such as ¹H NMR was used for experimentally confirming the prevalent template–monomer interactions derived from the modeling results. Molecular dynamics simulations indicating monomer dimerization in the prepolymerization solution correlated with the nature of the porogenic solvent, which was confirmed by NMR studies on hydrogen-bonding interactions of methacrylic acid in different solvents. Furthermore, batch rebinding studies revealed that the specific functionalities of the monomers essential to rebinding are retained after polymerization, which proves that the application of computational methods for modeling the prepolymerization solution provides useful information for optimizing real MIP systems.

Keywords: Molecularly imprinted polymers; 17β-Estradiol; Molecular modeling; ¹H NMR spectroscopy

Chiral effects of alkyl-substituted derivatives of N,O-bismethacryloyl ethanolamine on the performance of one monomer molecularly imprinted polymers (OMNiMIPs) by Jason LeJeune; David A. Spivak (pp. 433-440).

New monomers were synthesized and evaluated for their molecular imprinting performance by a recently discovered methodology referred to as one monomer molecularly imprinted polymers (OMNiMIPs). The structural design of the new monomers was based on a lead compound methacrylamidoethyl methacrylate (1) used for the synthesis of OMNiMIP1, and introduced alkyl groups of various sizes at the α-amino position of the lead compound. Enantioselectivity, determined by liquid chromatography, was used to compare the performance of the imprinted polymers. Methyl substitution provided crosslinker 5 (2-methacrylamidopropyl methacrylate), which upon imprint polymerization afforded OMNiMIP5 with approximately the same enantioselectivity (α = 3.8) as OMNiMP1 (α = 3.7) made with the lead compound (1). The other two monomers (6 and 7) with larger alkyl substitutions (isopropyl and sec-butyl respectively) resulted in OMNiMIPs with low selectivity values (α = 1.0 and 1.2 respectively). Last, a strong influence of diastereomeric complexes on OMNiMIP5 selectivity was determined, with L/L and D/D monomer/template pairs giving α values of 3.6–3.8, while L/D and D/L monomer/template pairs had α values of 2.3–2.4. There is no intrinsic enantioselectivity seen for the OMNiMIP5 control polymer made without template at all, giving an α value of 1.03.

Keywords: Molecular imprinting; Resolution; OMNiMIPs; Crosslinkers; Enantioselectivity; Chromatography

Molecularly imprinted sol–gel nanoparticles for mass-sensitive engine oil degradation sensing by Peter A. Lieberzeit; Adeel Afzal; Gerd Glanzing; Franz L. Dickert (pp. 441-446).

Titanate sol–gel layers imprinted with midchain carbonic acids have proven highly useful for detecting engine oil degradation processes owing to selective incorporation of oxidised base oil components. Synthesising the material from TiCl₄ in CCl₄ and precipitating with water leads to imprinted TiO₂ nanoparticles with a diameter of 200–300 nm. Replacing the water by a 1 M ammonium hydroxide solution reduces the average particle size to 50–100 nm with retention of the interaction capabilities. Experiments with the latter solution revealed that the 100-nm particles take up substantially more analyte, indicating a size-dependent phenomenon. As the number of interaction sites within each material is the same, this cannot be a consequence of thermodynamics but must be one of accessibility. The sensor characteristic of water-precipitated particles towards engine oil degradation products shows substantially increased sensitivity and dynamic range compared with the corresponding thin films. Coating quartz crystal microbalances with such nanoparticle materials leads to engine oil degradation sensors owing to incorporation of acidic base oil oxidation products. Interaction studies over a large range of layer thicknesses revealed that both the absolute signal and the steepness of the correlation between the sensor signal and the layer height is 2 times higher for the particles. Figure Generation of molecularly imprinted sol–gel nanoparticles

Keywords: Titanate sol–gel; Nanoparticles; Engine oil degradation sensor; Mass-sensitive detection; Sensitivity enhancement

‘Gate effect’ in templated polyacrylamide membranes influences the electrotransport of proteins and finds applications in proteome analysis by Alessandra Bossi; Matteo Andreoli; Francesca Bonini; Sergey Piletsky (pp. 447-454).

Templating is an effective way for the structural modifications of a material and hence for altering its functional properties. Here protein imprinting was exploited to alter polymeric polyacrylamide (PAA) membranes. The sieving properties and selection abilities of the material formed were evaluated by studying the electrically driven transport of various proteins across templated PAA membranes. The sieving properties correlated with the templating process and depended on the quantity of template used during the polymerisation. For 1 mg/mL protein-templated membranes a ‘gate effect’ was shown, which induced a preferential migration of the template and of similar-size proteins. Such template preferential electrotransport was exploited for the selective removal of certain proteins in biological fluids prior to proteome analysis (depletion of albumin from human serum); the efficiency of the removal was demonstrated by analysing the serum proteome by two-dimensional electrophoresis experiments. Figure PAA templeted membrane for the electroremoval of serum albumin before proteome analysis

Keywords: Molecularly imprinted polymers; Polyacrylamide membranes; Electrotransport; Human serum albumin depletion; Proteome

Application of the Doehlert experimental design to molecularly imprinted polymers: surface response optimization of specific template recognition as a function of the type and degree of cross-linking by Claire Rossi; Karsten Haupt (pp. 455-460).

We propose the use of Doehlert’s experimental design, a second-order uniform shell design, for the optimization of molecularly imprinted polymers (MIPs). We have chosen a simple model system where the influence of kind and degree of cross-linking on template recognition was studied using S-propranolol as the template. We found that Doehlert’s design allows—with very few experiments—one to screen the evolution of the binding capacity of a MIP as a function the different parameters, and thus appears to be a powerful means to screen for the best composition and synthesis method for MIPs. We believe that this chemometric tool can significantly accelerate the development of new MIPs as synthetic recognition elements, particularly in the context of a given application, and will be a versatile complement or alternative to first-order designs to fit complex processes.

Keywords: Chemometrics; Statistics; Molecularly imprinted polymers (MIP); Polymers

Co-immunoprecipitations revisited: an update on experimental concepts and their implementation for sensitive interactome investigations of endogenous proteins by Kelly Markham; Yu Bai; Gerold Schmitt-Ulms (pp. 461-473).

The study of protein–protein interactions involving endogenous proteins frequently relies on the immunoaffinity capture of a protein of interest followed by mass spectrometry-based identification of co-purifying interactors. A notorious problem with this approach is the difficulty of distinguishing physiological interactors from unspecific binders. Additional challenges pose the need to employ a strategy that is compatible with downstream mass spectrometry and minimizes sample losses during handling steps. Finally, the complexity of data sets demands solutions for data filtering. Here we present an update on co-immunoprecipitation procedures for sensitive interactome mapping applications. We define the relevant terminology, review methodological advances that reduce sample losses, and discuss experimental strategies that facilitate recognition of candidate interactors through a combination of informative controls and data filtering. Finally, we provide starting points for initial validation experiments and propose conventions for manuscripts which report on co-immunoprecipitation work.

Keywords: Co-immunoprecipitations; Mass spectrometry; Isotopic labeling; Antibodies; In vivo crosslinking

Advanced TMAH and TMAAc thermochemolysis–pyrolysis techniques for molecular characterization of size-separated fractions from aquatic dissolved organic matter by Juhani Peuravuori; Kalevi Pihlaja (pp. 475-491).

The structural similarities and differences between the original DOM and the eight size fractions separated were studied in detail with the pyrolysis technique in combination with gas chromatography and mass spectrometry (Py–GC–MS) using two alkylating reagents: TMAH (tetramethylammonium hydroxide), to find both esterified and free carboxylic acids; and TMAAc (tetramethylammonium acetate), to specify only free carboxylic acids. A statistical analysis of the original multidimensional TMAH and TMAAc pyrograms disclosed that the overall structural compositions of the five most important size fractions, accounting for 84% of the original DOM, greatly resembled each other. The remaining three minor size fractions were not classified as homogeneous associations, but they also contained the same total, covalently bound and free carboxylic acid species as the other size fractions and the original DOM mixture, thus representing some kind of intermediate forms. This fundamental outcome strongly supports the opinion that the native dissolved humic-like macromolecules resemble supramolecular associations of smaller molecular size moieties with similar structural functionalities. The concentrations of free aliphatic and aromatic dicarboxylic acids in the DOM solution were so low that their effects on the potential formation of multiply charged ions in electrospray ionization–MS (ESI–MS) studies are likely insignificant.

Keywords: Thermochemolysis; Carboxylic acids; Humic substances; Supramolecular associations; Molecular mixtures; Molecular sizes

A nanogold-quenched fluorescence duplex probe for homogeneous DNA detection based on strand displacement by Z.-H. Mo; X.-C. Yang; K.-P. Guo; Z.-Y. Wen (pp. 493-497).

A nanogold-quenched fluorescence duplex probe has been developed for lighting up homogenous hybridization assays. This novel probe is constructed from two strands of different lengths, and labeled by nanogold and a fluorophore at the long-strand 5′-end and the short-strand 3′-end, respectively. The two tags are in close contact, resulting in complete quenching of the probe fluorescence. If perfectly complemented to the nanogold-labeled strand, a long target oligonucleotide would displace the short fluorophore-labeled strand, and as a result, restore the fluorescence. By using nanogold in the probe, an extremely high quenching efficiency (99.1%) and removal of free fluorophore-labeled strand is achieved. The signal-to-noise ratio and the detection limit (50 pmol L⁻¹) of homogenous assays are therefore improved significantly, in comparison with similar probes using organic acceptors. Moreover, the probe has a great inhibition effect on hybridization to a mismatched oligonucleotide. This effect provides the assay with a high specificity, and particularly the assay has great potential in applications for discriminating variations in sequences. The assay sensitivity could be markedly enhanced by using fluorescent materials in the signal strand that are brighter and not quenched by nucleobases.

Keywords: DNA detection; Fluorescence quenching; Nanogold; Displacement hybridization

Optimized carbon nanotube fiber microelectrodes as potential analytical tools by Lucie Viry; Alain Derré; Patrick Garrigue; Neso Sojic; Philippe Poulin; Alexander Kuhn (pp. 499-505).

The preparation and interesting electrochemical properties of carbon nanotube (CNT) fiber microelectrodes are reported. By combining the advantages of CNT with those of fiber electrodes, this type of microelectrode differs from CNT-modified or CNT-containing composite electrodes, because they are made solely of CNT without other components, for example additives or binders. The performance of these electrodes has been characterized with regard to, among others, the electrocatalytic oxidation of analytes via dehydrogenase-mediated reactions. In this context the reversible regeneration of the coenzyme NAD⁺ using a mediator is a key step in the development of new amperometric sensor devices and we have successfully immobilized mediator molecules that are very efficient for this purpose on the surface of the CNT fiber electrode. The microelectrodes thus obtained have been compared with classic carbon microelectrodes and have promising behavior in biosensing applications, especially after specific pretreatments such as CNT alignment inside the fiber or expansion of the specific surface by chemically induced swelling.

Keywords: Carbon nanotubes; Microelectrodes; Electrocatalysis; NADH oxidation; Glucose sensor

Development of automated amperometric detection of antibodies against Bacillus anthracis protective antigen by Zoraida P. Aguilar; Mali Sirisena (pp. 507-515).

Picogram levels of antibodies against the protective antigen (PA) of Bacillus anthracis were detected in an automated electrochemical sandwich-type enzyme-linked immunosorbant assay. The antibodies were captured and detected using an 8 × 3 array of 50-μm-diameter cavities. The reagent and sample volumes were as low as 200 nL in a less than 25-min assay from capture to signal generation. The electrochemical detection of the antibodies was demonstrated at 0.05–10 μg/mL containing only 10–5,000 pg antibodies. The limit of detection is 10 fg for a 200-nL sample. Detection of anti-PA immunoglobulin G performed in spiked normal human serum and fresh whole human blood did not show a significant difference from detection in a buffer. The initial automation of the assay involved the use of a digital syringe pump for the delivery of reagents to the capture surface.

Keywords: Automated; Amperometric; Bacillus anthracis ; Self-contained microelectrodes; Immunoassay

Capacitive biosensor for detection of endotoxin by Warakorn Limbut; Martin Hedström; Panote Thavarungkul; Proespichaya Kanatharana; Bo Mattiasson (pp. 517-525).

A capacitive biosensor for the detection of bacterial endotoxin has been developed. Endotoxin-neutralizing protein derived from American horseshoe crab was immobilized to a self-assembled thiol layer on a biosensor transducer (Au). Upon injection of a sample containing endotoxin, a decrease in the observed capacitive signal was registered. Endotoxin could be determined under optimum conditions with a detection limit of 1.0 × 10⁻¹³ M and linearity ranging from 1.0 × 10⁻¹³ to 1.0 × 10⁻¹⁰ M. Good agreement was achieved when applying endotoxin preparations purified from an Escherichia coli cultivation to the capacitive biosensor system, utilizing the conventional method for quantitative endotoxin determination, the Limulus amebocyte lysate test as a reference. The capacitive biosensor method was statistically tested with the Wilcoxon signed rank test, which proved the system is acceptable for the quantitative analysis of bacterial endotoxin (P < 0.05). Figure The flow-injection capacitive biosensor system and the capacitive properties of the transducer surface, where C_SAM is the capacitance change of the self-assembled thiol monolayer, C_P is the capacitance change of the protein layer, C_a is the capacitance change of the analyte layer and C_Total is the total capacitance change measured at the working electrode/solution interface (modified from Limbut et al., 2006. Biosens Bioelectron 22: 233-240)

Keywords: Capacitive biosensor; Endotoxin; Limulus ; Self-assembled monolayer; Monolayer

Room temperature ionic liquid doped DNA network immobilized horseradish peroxidase biosensor for amperometric determination of hydrogen peroxide by Cunlan Guo; Yonghai Song; Hui Wei; Peicai Li; Li Wang; Lanlan Sun; Yujing Sun; Zhuang Li (pp. 527-532).

A novel electrochemical H₂O₂ biosensor was constructed by embedding horseradish peroxide (HRP) in a 1-butyl-3-methylimidazolium tetrafluoroborate doped DNA network casting on a gold electrode. The HRP entrapped in the composite system displayed good electrocatalytic response to the reduction of H₂O₂. The composite system could provide both a biocompatible microenvironment for enzymes to keep their good bioactivity and an effective pathway of electron transfer between the redox center of enzymes, H₂O₂ and the electrode surface. Voltammetric and time-based amperometric techniques were applied to characterize the properties of the biosensor. The effects of pH and potential on the amperometric response to H₂O₂ were studied. The biosensor can achieve 95% of the steady-state current within 2 s response to H₂O₂. The detection limit of the biosensor was 3.5 μM, and linear range was from 0.01 to 7.4 mM. Moreover, the biosensor exhibited good sensitivity and stability. The film can also be readily used as an immobilization matrix to entrap other enzymes to prepare other similar biosensors. Figure Horseradish peroxidase (HRP) embedded in a 1-butyl-3-methylimidazolium tetrafluoroborate (BMIM·BF 4) doped DNA network can be used to fabricate a HRP sensor for the determination of H₂O₂

Keywords: DNA; Room temperature ionic liquid; Horseradish peroxidase; Hydrogen peroxide; Biosensor

Enzymatic detection of glucose using fluoride-selective electrodes with polymeric membranes by Łukasz Górski; Dorota Klimaszewska; Mariusz Pietrzak; Elżbieta Malinowska (pp. 533-539).

The feasibility of using polymeric membrane fluoride-selective electrodes based on zirconium(IV) 5,10,15,20-tetraphenylporphyrin as a detector in a flow-injection analysis (FIA) system for glucose determination was examined. The optimization of enzymatic reactions, FIA system configuration and enzyme-immobilization process was performed. It was shown that the resulting flow-injection system exhibits good working parameters, such as reproducibility, linear range of glucose concentration (3 × 10⁻³–10⁻¹ M), sampling rate (60 samples per minute) and lifetime (over 1 month). The performance of the polymeric membrane electrode was similar to that of a crystalline LaF₃ electrode. The results of glucose determination in synthetic samples with the proposed system show good agreement with real glucose concentrations.

Keywords: Ion-selective electrodes; Biosensors; Chemical sensors; Enzymes; Flow injection

Comparative study of different approaches to the determination of robustness for a sensitive-stacking capillary electrophoresis method. Estimation of system suitability test limits from the robustness test by Teresa Galeano-Díaz; María-Isabel Acedo-Valenzuela; Nielene Mora-Díez; Antonio Silva-Rodríguez (pp. 541-553).

A robustness study for a sensitive-stacking capillary electrophoresis method based on “acetonitrile-stacking” was carried out. Ten variables (pH, acetonitrile and triethanolamine in the buffer, injection time, injection pressure, acetonitrile and NaCl in the sample, capillary and tray temperature and separation voltage), whose levels were varied by 10% around the nominal level, were examined by a Plackett–Burman design (two-level design). The effects on corrected peak area and resolution (responses) were calculated and interpreted using three statistical approaches: dummy variables, distribution effects (Dong’s algorithm) and calibration curve. Dong’s method was found to be the most suitable to evaluate the robustness, since it considers qualitative (resolution) and quantitative (corrected peak area) responses and does not need a minimum number of dummy variables in the experimental design. From these studies, we can deduce that the first four variables were significant at 10% around the nominal level, and therefore a new design was made with those four variables at 5% nominal level. Then, only two variables proved to be significant for the resolution between some peaks, so the system suitability test limits were defined for these resolutions.

Keywords: Robustness test; Sensitive-stacking capillary electrophoresis method; Plackett–Burman design; System suitability test limits

A new conceptual approach to assigning biomass combustion-derived methoxyphenol structures by using a gas chromatographic retention index model by S. Rayne; N. J. Eggers (pp. 555-561).

A new conceptual approach towards iteratively constructing chromatographic retention time/index models is presented. The approach is applicable where there is potential structural uncertainty in a number of members of the dataset used in constructing the model, and where limited spectroscopic information is available to guide the process. The model is demonstrated on a suite of biomass combustion-derived methoxyphenols for which gas chromatographic polydimethylsiloxane retention index data was available in the literature, but where there was ambiguity regarding the identity of several members of the dataset. The retention property model is populated by sequentially screening a series of candidate structures that meet basic mass spectrometric requirements by using a multiple linear regression model containing molecular and physicochemical properties that have been previously shown to yield reliable predictions of chromatographic behaviour within a compound class. The criteria for deciding on the likely structure(s) out of a suite of candidate structures is based upon the improved quality of fit the most probable structure gives the regression model relative to other candidate structures.

Keywords: Chromatographic retention; Prediction; Guaiacyls; Methoxyphenols; Structural determination

Pipette tip solid-phase extraction and gas chromatography – mass spectrometry for the determination of methamphetamine and amphetamine in human whole blood by Chika Hasegawa; Takeshi Kumazawa; Xiao-Pen Lee; Akemi Marumo; Natsuko Shinmen; Hiroshi Seno; Keizo Sato (pp. 563-570).

Methamphetamine and amphetamine were extracted from human whole blood samples using pipette tip solid-phase extraction (SPE) with MonoTip C₁₈ tips, on which C₁₈-bonded monolithic silica gel was fixed. Human whole blood (0.1 mL) containing methamphetamine and amphetamine, with N-methylbenzylamine as an internal standard, was mixed with 0.4 mL of distilled water and 50 μL of 5 M sodium hydroxide solution. After centrifugation, the supernatant was extracted to the C₁₈ phase of the tip (pipette tip volume, 200 μL) by 25 repeated aspirating/dispensing cycles using a manual micropipettor. Analytes retained in the C₁₈ phase were eluted with methanol by five repeated aspirating/dispensing cycles. After derivatization with trifluoroacetic anhydride, analytes were measured by gas chromatography – mass spectrometry with selected ion monitoring in the positive-ion electron impact mode. Recoveries of methamphetamine and amphetamine spiked into whole blood were more than 87.6 and 81.7%, respectively. Regression equations for methamphetamine and amphetamine showed excellent linearity in the range of 0.5–100 ng/0.1 mL. The limits of detection for methamphetamine and amphetamine were 0.15 and 0.11 ng/0.1 mL, respectively. Intra- and interday coefficients of variation for both stimulants were not greater than 9.6 and 13.8%, respectively. The determination of methamphetamine and amphetamine in autopsy whole blood samples is presented, and was shown to validate the present methodology.

Keywords: Solid-phase extraction (SPE); Methamphetamine; Amphetamine; Gas chromatography; Mass spectrometry

Simultaneous determination of bioactive constituents in Danggui Buxue Tang for quality control by HPLC coupled with a diode array detector, an evaporative light scattering detector and mass spectrometry by Ling Yi; Lian-Wen Qi; Ping Li; Yi-Han Ma; Yong-Jing Luo; Hai-Yun Li (pp. 571-580).

Danggui Buxue Tang (DBT), a classical traditional Chinese formula comprising Radix Angelicae Sinensis (RAS) and Radix Astragali (RA), has been widely used to treat menopausal irregularity in Chinese women for nearly 800 years. In this study, a comprehensive analytical method of simultaneously determining the main types of bioactive constituents, eighteen in all from the formula, involving flavonoids, saponins, organic acid and some volatile compounds, was developed. This method was based on HPLC coupled to a diode array and evaporative light scattering detectors (HPLC–DAD–ELSD) on a common reverse-phase C₁₈ column. Liquid chromatography coupled with on-line electrospray ionization mass spectrometry (LC–ESI–MS) was also used to further validate and analyze the constituents. It was found that 0.3% aqueous formic acid and acetonitrile was the optimum mobile phase for gradient elution. This method, which showed good precision and accuracy, was successfully used to quantify the bioactive constituents in six products. As a result, the validated HPLC method, together with the LC–ESI–MS analysis, provided a new basis for assessing the quality of traditional Chinese medicinal compound preparations (TCMCPs) consisting of many bioactive components. Figure 1 Products of Danggui Buxue Tang Figure 2 HPLC Chromatograms of Eighteen Reference Standards

Keywords: Danggui Buxue Tang; Bioactive constituents; Quality control; HPLC–DAD–ELSD; LC–ESI–MS

Development of a novel immunobiosensor method for the rapid detection of okadaic acid contamination in shellfish extracts by Nuria M. Llamas; Linda Stewart; Terry Fodey; H. Cowan Higgins; María Luisa R. Velasco; Luis M. Botana; Christopher T. Elliott (pp. 581-587).

The mouse bioassay is the methodology that is most widely used to detect okadaic acid (OA) in shellfish samples. This is one of the best-known toxins, and it belongs to the family of marine biotoxins referred to as the diarrhetic shellfish poisons (DSP). Due to animal welfare concerns, alternative methods of toxin detection are being sought. A rapid and specific biosensor immunoassay method was developed and validated for the detection of OA. An optical sensor instrument based on the surface plasmon resonance (SPR) phenomenon was utilised. A polyclonal antibody to OA was raised against OA–bovine thyroglobulin conjugate and OA–N-hydroxy succinimide ester was immobilised onto an amine sensor chip surface. The assay parameters selected for the analysis of the samples were: antibody dilution, 1/750; ratio of antibody to standard, 1:1; volume of sample injected, 25 μl min⁻¹; flow rate, 25 μl min⁻¹. An assay action limit of 126 ng g⁻¹ was established by analysing of 20 shellfish samples spiked with OA at the critical concentration of 160 ng g⁻¹, which is the action limit established by the European Union (EU). At this concentration of OA, the assay delivered coefficient of variations (CVs) of <10%. The chip surface developed was shown to be highly stable, allowing more than 50 analyses per channel. When the concentrations of OA determined with the biosensor method were compared with the values obtained by LC–MS in contaminated shellfish samples, the correlation between the two analytical methods was found to be highly satisfactory (r ² = 0.991). Figure Biacore

Keywords: Biosensors; Okadaic acid; Diarrhetic shellfish poisons; Mouse bioassay

Migration of organic compounds from a multilayer plastic–paper material intended for food packaging by Cristina Nerín; Esther Asensio (pp. 589-596).

This paper deals with the study of the kinetic migration of a series of organic compounds representative of potential contaminants in packaging materials and used as surrogates (o-xylene, acetophenone, dodecane, naphthalene, 2,3,4-trichloroanisole, benzophenone, isomeric mix of diisopropylnaphthalene, diisobutyl phthalate and methyl stearate). Migration to one side of a solid simulant, Tenax®, also referred to as MPPO (modified polyphenylene oxide), was investigated in this study. One spiked sample of multilayer material was used to optimise the extraction procedures for the multilayer paper-based material and the Tenax as well as to perform kinetic migration studies. Three sequential extractions using ethanol were necessary for the strips of the multilayer material but only one extraction was necessary for the solid simulant to obtain >70% recovery of the surrogates. These results enabled us to specify the extraction requirements of the multilayer sample and the solid simulant and as well as those of the migration tests at high temperature using Tenax as solid simulant. The matrix effect associated with the extraction of the contaminants from the multilayer sample is also discussed.

Keywords: Food packaging; Kinetic migration tests; Multilayer; Tenax®; Surrogates; Paper; Polyethylene

Stable carbon isotope ratios of methyl-branched fatty acids are different to those of straight-chain fatty acids in dairy products by Walter Vetter; Simon Gaul; Saskia Thurnhofer; Kathrin Mayer (pp. 597-604).

Methyl-branched fatty acids (MBFAs) are the dominant form of fatty acid found in many bacteria. They are also found at low levels in a range of foodstuffs, where their presence has been linked to bacterial sources. In this study we evaluated the potential of compound-specific isotope analysis to obtain insights into the stable carbon isotope ratios (δ¹³C values in ‰) of individual MBFAs and to compare them to the stable carbon isotope ratios of straight-chain fatty acids in food. Due to their low abundance in foodstuffs, the MBFAs were enriched prior to gas chromatography coupled to isotope ratio mass spectrometric (GC–IRMS) analysis. After transesterification, urea complexation was used to suppress the 16:0 and 18:0 methyl esters that were dominant in the samples. Following that, silver-ion high performance liquid chromatography was used to separate the saturated from the unsaturated fatty acids. The resulting solutions of saturated fatty acids obtained from suet, goat’s milk, butter, and human milk were studied by GC–IRMS. The δ¹³C values of fatty acids with 12–17 carbons ranged from −25.4‰ to −37.6‰. In all samples, MBFAs were most depleted in carbon-13, followed by the odd-chain fatty acids 15:0 and 17:0. 14:0 and 16:0 contained the highest proportions of carbon-13. The results from this study illustrate that MBFAs have distinctive δ¹³C values and must originate from other sources and/or from very different substrates. These measurements support the initial hypothesis that δ¹³C values can be used to attribute MBFAs to particular sources.

Keywords: Dairy products; Fatty acids; Branched-chain fatty acids; Stable carbon isotopes; Compound-specific isotope analysis; Isotope ratio mass spectrometry

Determination of 2-isopropyl thioxanthone and 2-ethylhexyl-4-dimethylaminobenzoate in milk: comparison of gas and liquid chromatography with mass spectrometry by Ana Gil-Vergara; Cristina Blasco; Yolanda Picó (pp. 605-617).

Liquid chromatography–tandem mass spectrometry (LC-MS/MS) and gas chromatography–mass spectrometry (GC-MS) have been compared for the analysis of 2-isopropyl thioxanthone (ITX) and 2-ethylhexyl-4-dimethylaminobenzoate (EHDAB). Pressurized liquid extraction (PLE) was applied for the extraction of ITX and EHDAB from milk and milk-based beverages. Samples were homogenized with sea sand and anhydrous sodium sulfate, and were extracted with ethyl acetate at 100 °C and 10.3 × 10⁶ Pa in one cycle of 10 min at 90% flush. Both, GC-MS and LC-MS/MS were suitable to determine these photoinitiators in the PLE extracts, providing appropriate identification and quantification. The recoveries obtained ranged from 70 to 99% for ITX and from 70 to 95% for EHDAB. These recoveries were equal as those obtained by a conventional liquid–liquid partitioning with acetonitrile and tert-butyl methyl ether–hexane. The quantification limits using GC-MS, based on a signal-to-noise ratio of 10, were 0.5 μg/L for ITX and 1 μg/L for EHDAB. The repeatability of the method, as indicated by the relative standard deviations, was within the range 0.9–16.1%. The same parameters calculated using LC-MS/MS result in quantification limits of 0.1 μg/L for ITX and 0.02 μg/L for EHDAB and repeatability within the range 5.2–19.4%. These results pointed out that both techniques are appropriate to determine these compounds in food samples. The method was applied to milk and milk-based beverages from different supermarkets. The ITX and EHDAB contents ranged from 2.5 to 325 μg/L and from 8 to 126 μg/L, respectively.

Keywords: 2-Isopropyl thioxanthone; 2-Ethylhexyl-4-dimethylaminobenzoate; Gas chromatography–mass spectrometry; Liquid chromatography–tandem mass spectrometry; Food analysis; Milk

Response surface methodology for the microwave-assisted extraction of insecticides from soil samples by M. Carmen Hernández-Soriano; Aránzazu Peña; M. Dolores Mingorance (pp. 619-630).

The extraction of two pyrethroid insecticides (deltamethrin and α-cypermethrin) together with three organophosphorus insecticides (dimethoate, diazinon and malathion) from soil samples was carried out with microwave-assisted technology. Experimental designs showed that extraction temperature, addition of water to the extractant and solvent/soil ratio were the variables that affected the recoveries of the pesticide the most. Response surface methodology was applied to find the optimum values of the variables involved in the extractions of the analytes. In addition, in order to achieve near-optimal extraction conditions, a desirability function was used to optimize the five pesticides simultaneously. The optimized conditions were applied to different types of soils.

Keywords: Microwave-assisted extraction; Soil samples; Pyrethroid pesticides; Organophosphorous pesticides; Box–Behnken design; Desirability function

Relative characterization of rosemary samples according to their geographical origins using microwave-accelerated distillation, solid-phase microextraction and Kohonen self-organizing maps by N. Tigrine-Kordjani; F. Chemat; B. Y. Meklati; L. Tuduri; J. L. Giraudel; M. Montury (pp. 631-641).

For centuries, rosemary (Rosmarinus officinalis L.) has been used to prepare essential oils which, even now, are highly valued due to their various biological activities. Nevertheless, it has been noted that these activities often depend on the origin of the rosemary plant and the method of extraction. Since both of these quality parameters can greatly influence the chemical composition of rosemary oil, an original analytical method was developed where “dry distillation” was coupled to headspace solid-phase microextraction (HS-SPME) and then a data mining technique using the Kohonen self-organizing map algorithm was applied to the data obtained. This original approach uses the newly described microwave-accelerated distillation technique (MAD) and HS-SPME; neither of these techniques require external solvent and so this approach provides a novel “green” chemistry sampling method in the field of biological matrix analysis. The large data set obtained was then treated with a rarely used chemometric technique based on nonclassical statistics. Applied to 32 rosemary samples collected at the same time from 12 different sites in the north of Algeria, this method highlighted a strong correlation between the volatile chemical compositions of the samples and their origins, and it therefore allowed the samples to be grouped according to geographical distribution. Moreover, the method allowed us to identify the constituents that exerted the most influence during classification.

Keywords: Rosemary essential oil; Dry distillation; HS-SPME; KSOMS; Data mining

Simplified multiresidue method for determination of pesticide residues in lettuce by gas chromatography with nitrogen–phosphorus detection by José Fenoll; Pilar Hellín; Josefa López; Alberto González; Pilar Flores (pp. 643-651).

A rapid and simple method has been developed for simultaneous determination of different classes of pesticide in different varieties of lettuce (Lactuca sativum). Lettuce samples were extracted by homogenization with acetone and partitioned into ethyl acetate–cyclohexane. Subsequent sample clean-up was not needed. Pesticide residues were determined by capillary gas chromatography with nitrogen–phosphorus detection (NPD). Confirmatory analysis of the pesticides was performed by capillary gas chromatography coupled with mass spectrometry in selected-ion-monitoring (SIM) mode. Recovery at two levels of fortification (ca. 0.05 and 0.20 mg kg⁻¹) ranged from 63.9 to 118.6%, and relative standard deviations were below 9.5%. The proposed method was used to determine pesticide levels in different types of lettuce grown in soil from experimental fields.

Keywords: Multiresidue; Lettuce; Pesticides; Gas chromatography

Applicability of a liquid membrane in enrichment and determination of nickel traces from natural waters by F. C. Domínguez-Lledó; María D. Galindo-Riaño; I. C. Díaz-López; M. García-Vargas; M. D. Granado-Castro (pp. 653-659).

In this work, a bulk liquid membrane method has been applied for Ni enrichment and separation from natural waters. The carrier-mediated transport was accomplished by pyridine-2-acetaldehyde benzoylhydrazone dissolved in toluene as a complexing agent. The preconcentration was achieved through pH control of source and receiving solutions via a counterflow of protons. The main variables were optimized by using a modified simplex technique. High transport efficiencies (101.2 ± 1.8–99.7 ± 4.2%) were provided by the carrier for nickel ions in a receiving phase of 0.31 mol L⁻¹ nitric acid after 9–13 h depending on sample salinity. The precision of the method was 2.05% (without a saline matrix) and 4.04% (with 40 g L⁻¹ NaCl) at the 95% confidence level and the detection limit of the blank was 0.015 μg L⁻¹ Ni for detection by atomic absorption spectroscopy. The applicability of the method was tested on certified reference and real water samples with successful results, even for saline samples. The relative errors were −0.60% for certified reference materials and ranged from −0.39 to 2.90% and from 0.3 to 11.05% for real samples, obtained by comparison of inductively coupled plasma mass spectrometry and adsorptive cathodic stripping voltammetry measurements, respectively.

Keywords: Nickel; Natural water; Liquid membrane; Preconcentration; Pyridine-2-acetaldehyde benzoylhydrazone

Preparation and certification of arsenobetaine reference material NMIJ CRM 7901-a by Tomohiro Narukawa; Koichi Chiba; Takayoshi Kuroiwa; Takashi Yarita; Akiko Takatsu (pp. 661-666).

An arsenobetaine [(CH₃)₃As⁺CH₂COO⁻] solution reference material, NMIJ CRM 7901-a, intended for use in the speciation of arsenic compounds, was developed and certified by the National Metrology Institute of Japan (NMIJ), part of the National Institute of Advanced Industrial Science and Technology (AIST). The high-purity arsenobetaine powder was synthesized from trimethylarsine [(CH₃)₃As], and it was dissolved in water in order to prepare 20 mg kg⁻¹ of arsenobetaine standard solution. The solution was bottled in 500 bottles (each containing 10 ml). Certification of the CRM for arsenobetaine was conducted by NMIJ. The concentration of As was determined by four independent analytical techniques (ICP–MS, ICP–OES, GFAAS and LC–ICP–MS), and each result was converted to the arsenobetaine concentration by applying an appropriate factor. The arsenobetaine concentration in the CRM was thus certified.

Keywords: Certification; Certified reference material; Arsenobetaine; Arsenic compounds; Speciation

Study of the feasibility of distinguishing cigarettes of different brands using an Adaboost algorithm and near-infrared spectroscopy by Chao Tan; Menglong Li; Xin Qin (pp. 667-674).

The feasibility of utilizing an Adaboost algorithm in conjuction with near-infrared (NIR) spectroscopy to automatically distinguish cigarettes of different brands was explored. Simple linear discriminant analysis (LDA) was used as the base algorithm to train all weak classifiers in Adaboost. Both principal component analysis (PCA) and its kernel version (kernel principal component analysis, KPCA) were used for feature extraction and were also compared to each other. The influence of the training set size on the final classification model was also investigated. Using a case study, it was demonstrated that Adaboost coupled with PCA or KPCA can obviously improve the ability to discriminate between samples that cannot be separated by a single linear classifier. However, in term of the overall performance, KPCA appears preferable to PCA for feature extraction, especially when the samples used for training are relatively small. The results also indicate that more training samples should be applied, if possible, in order to fully demonstrate the superiority of Adaboost. It seems that the use of an Adaboost algorithm in conjunction with NIR spectroscopy in combination with KPCA for feature extraction comprises a promising tool for distinguishing cigarettes of different brands, especially in situations where there is an obvious overlap between the NIR spectra afforded by cigarettes of different brands.

Keywords: Near-infrared; Cigarette; Identification; Adaboost

Sections

Personal tools

Analytical and Bioanalytical Chemistry (v.389, #2)

Sections

Personal tools

Local resources

Analytical and Bioanalytical Chemistry (v.389, #2)