Analytica Chimica Acta (v.610, #1)

Contents (v-vi).

A review on development of solid phase microextraction fibers by sol–gel methods and their applications by Ashwini Kumar; Gaurav; Ashok Kumar Malik; Dhananjay Kumar Tewary; Baldev Singh (1-14).
Solid phase microextraction (SPME) is an innovative, solvent free technology that is fast, economical and versatile. SPME is a fiber coated with a liquid (polymer), a solid (sorbent) or a combination of both. The fiber coating takes up the compounds from the sample by absorption in the case of liquid coatings or adsorption in the case of solid coatings. The SPME fiber is then transferred with the help of a syringe like device into the analytical instrument for desorption and analysis of the target analytes. The sol–gel process provides a versatile method to prepare size, shape and charge selective materials of high purity and homogeneity by means of preparation techniques different from the traditional ones, for the chemical analysis. This review is on the current state of the art and future trends in the developments of solid phase microextraction (SPME) fibers using sol–gel method. To achieve more selective determination of different compound classes, the variety of different coating material for SPME fibers has increased. Further developments in SPME as a highly efficient extraction technique, will greatly depend on new breakthroughs in the area of new coating material developments for the SPME fibers. In sol–gel approach, appropriate sol–gel precursors and other building blocks can be selected to create a stationary phase with desired structural and surface properties. This approach is efficient in integrating the advantageous properties of organic and inorganic material systems and thereby increasing and improving the extraction selectivity of the produced amalgam organic–inorganic stationary phases. This review is mainly focused on recent advanced developments in the design, synthesis, characterisation, properties and application of sol–gel in preparation of coatings for the SPME fibers.
Keywords: Sol–gel technology; Solid phase microextraction; High pressure liquid chromatography; Gas chromatography; Capillary electrophoresis;

Spectroscopic imaging is well suited to the study of micro-samples from artworks, where the sample material is limited and the maximum amount of information needs to be obtained. In this study, a new approach to imaging elemental data from energy dispersive X-ray analysis maps was used in conjunction with micro-Raman spectroscopic imaging to characterise the paint layers within micro-samples. Cross-sections from the 16th century painting Portrait of a Youth were found to contain vermilion, lead–tin yellow type 1 and a blue-green pigment consistent with terre-verte. The mid-preparatory layer (imprimatura) contains a high proportion of elements and mineral inclusions that indicates a clay-type composition. The ground layer was identified as anhydrite with large gypsum inclusions. The pigments and composition of the preparatory layers are consistent with those used by Italian Renaissance artist Dosso Dossi.
Keywords: Multispectral; Hyperspectral; Image analysis; Paintings; Cross-sections; Renaissance art;

A quantitative structure–property relation (QSPR) study was conducted on the solubility in supercritical fluid carbon dioxide (SCF-CO2) of some recently synthesized anthraquinone, anthrone and xanthone derivatives. The data set consisted of 29 molecules in various temperatures and pressures, which form 1190 solubility data. The combined data splitting-feature selection (CDFS) strategy, which previously developed in our research group, was used as descriptor selection and model development method. Modeling of the relationship between selected molecular descriptors and solubility data was achieved by linear (multiple linear regression; MLR) and nonlinear (artificial neural network; ANN) methods. The QSPR models were validated by cross-validation as well as application of the models to predict the solubility of three external set compounds, which did not have contribution in model development steps. Both linear and nonlinear methods resulted in accurate prediction whereas more accurate results were obtained by ANN model. The respective root mean square error of prediction obtained by MLR and ANN models were 0.284 and 0.095 in the term of logarithm of g solute m−3 of SCF-CO2. A comparison was made between the models selected by CDFS method and the conventional stepwise feature selection method. It was found that the latter produced models with higher number of descriptors and lowered prediction ability, thus it can be considered as an over-fitted model.
Keywords: Solubility; Supercritical carbon dioxide; Quantitative structure–property relationship; Combined data splitting-feature selection; Anthraquinone;

Extraction of domoic acid from seawater and urine using a resin based on 2-(trifluoromethyl)acrylic acid by Elena V. Piletska; Fernando Navarro Villoslada; Iva Chianella; Alessandra Bossi; Kal Karim; Michael J. Whitcombe; Sergey A. Piletsky; Gregory J. Doucette; John S. Ramsdell (35-43).
A new solid-phase extraction (SPE) matrix with high affinity for the neurotoxin domoic acid (DA) was designed and tested. A computational modelling study led to the selection of 2-(trifluoromethyl)acrylic acid (TFMAA) as a functional monomer capable of imparting affinity towards domoic acid. Polymeric adsorbents containing TFMAA were synthesised and tested in high ionic strength solutions such as urine and seawater. The TFMAA-based polymers demonstrated excellent performance in solid-phase extraction of domoic acid, retaining the toxin while salts and other interfering compounds such as aspartic and glutamic acids were removed by washing and selective elution. It was shown that the TFMAA-based polymer provided the level of purification of domoic acid from urine and seawater acceptable for its quantification by high performance liquid chromatography–mass spectrometry (HPLC–MS) and enzyme-linked immunosorbent assay (ELISA) without any additional pre-concentration and purification steps.
Keywords: Domoic acid; Solid-phase extraction; High performance liquid chromatography–mass spectrometry; Enzyme-linked immunosorbent assay;

A potential high-throughput method for the determination of lipase activity by potentiometric flow injection titrations by Katja Vahl; Heike Kahlert; Dominique Böttcher; Rainer Wardenga; Šebojka Komorsky-Lovrić; Uwe Bornscheuer; Fritz Scholz (44-49).
Potentiometric FIA titrations were performed to determine enzyme activities of lipase type B from Candida antarctica, CAL-B. Two substrates, triacetin and tributyrin were hydrolyzed in phosphate buffer solutions, and the concentration change of the base component of the buffer was titrated in a carrier solution containing hydrochloric acid and potassium chloride. The system was calibrated with butyric acid and acetic acid, respectively. FIA titration peaks were evaluated with respect to peak height and peak area. Butyric acid and acetic acid could be titrated in the buffer solution from 3 × 10−3  mol L−1 to 0.1 mol L−1. The detection limit of enzyme activity was determined to be 0.07 U mL−1 (15 min reaction time) and the minimum activity was calculated to be 0.035 units corresponding to 35 nmol min−1. The specific activities of lipase B for the hydrolysis of tributyrin and triacetin were determined as 16 ± 2 U mg−1 and 2 ± 0.2 U mg−1 (per mg commercial lipase preparation), respectively.
Keywords: Flow injection titrations; High-throughput screening; Lipase activity; Composite pH-detector;

Cadmium telluride quantum dots as pH-sensitive probes for tiopronin determination by Yun-Qing Wang; Chao Ye; Zheng-Hui Zhu; Yu-Zhu Hu (50-56).
The pH-sensitive cadmium telluride (CdTe) quantum dots (QDs) were used as proton probes for tiopronin determination. Based on the fluorescence quenching of CdTe QDs caused by tiopronin, a simple, rapid and specific quantitative method was proposed. Under the optimal conditions, the calibration plot of ln(F 0/F) with concentration of tiopronin was linear in the range of 0.15–20 μg mL−1(0.92–122.5 μmol L−1) with correlation coefficient of 0.998. The limit of detection (LOD) (3σ/k) was 0.15 μg mL−1(0.92 μmol mL−1). The content of tiopronin in pharmaceutical tablet was determined by the proposed method and the result agreed with that obtained from the oxidation–reduction titration method and the claimed value.
Keywords: Cadmium telluride; pH sensitive; Fluorescence quenching; Tiopronin; Determination;

Previous studies have indicated that nitrated explosives may be detected by fluorescence quenching of pyrene and related compounds. The use of pyrene, however, invokes numerous health and waste disposal hazards. In the present study, ten safer fluorophores are identified for quenching detection of target nitrated compounds. Initially, Stern–Volmer constants are measured for each fluorophore with nitrobenzene and 4-nitrotoluene to determine the sensitivity of the quenching interaction. For quenching constants greater than 50 M−1, sensitivity and selectivity are investigated further using an extended set of target quenchers. Nitromethane, nitrobenzene, 4-nitrotoluene, and 2,6-dinitrotoluene are chosen to represent nitrated explosives and their degradation products; aniline, benzoic acid, and phenol are chosen to represent potential interfering compounds. Among the fluorophores investigated, purpurin, malachite green, and phenol red demonstrate the greatest sensitivity and selectivity for nitrated compounds. Correlation of the quenching rate constants for these fluorophores to Rehm–Weller theory suggests an electron-transfer quenching mechanism. As a result of the large quenching constants, purpurin, malachite green, and phenol red are the most promising for future detection of nitrated explosives via fluorescence quenching.
Keywords: Fluorescence; Fluorescence quenching; Stern–Volmer constant; Rehm–Weller quenching theory; Explosives;

Influence of quantum dot's quantum yield to chemiluminescent resonance energy transfer by Hai-Qiao Wang; Yong-Qiang Li; Jian-Hao Wang; Qiao Xu; Xiu-Qing Li; Yuan-Di Zhao (68-73).
The resonance energy transfer between chemiluminescence donor (luminol–H2O2 system) and quantum dots (QDs, emission at 593 nm) acceptors (CRET) was investigated. The resonance energy transfer efficiencies were compared while the oil soluble QDs, water soluble QDs (modified with thioglycolate) and QD–HRP conjugates were used as acceptor. The fluorescence of QD can be observed in the three cases, indicating that the CRET occurs while QD acceptor in different status was used. The highest CRET efficiency (10.7%) was obtained in the case of oil soluble QDs, and the lowest CRET efficiency (2.7%) was observed in the QD–HRP conjugates case. This result is coincident with the quantum yields of the acceptors (18.3% and 0.4%). The same result was observed in another similar set of experiment, in which the amphiphilic polymer modified QDs (emission at 675 nm) were used. It suggests that the quantum yield of the QD in different status is the crucial factor to the CRET efficiency. Furthermore, the multiplexed CRET between luminol donor and three different sizes QD acceptors was observed simultaneously. This work will offer useful support for improving the CRET studies based on quantum dots.
Keywords: Luminol; Quantum dots; Quantum yield; Chemiluminescence resonance energy transfer;

Two holy water fonts (dated at the beginning of the XVII century) in the Santa Maria della Steccata Church in Parma (Italy) have recently been restored. Before the intervention, a detailed investigation on their degradation products was carried out to understand the mechanisms of alteration of the materials and to suggest appropriate restoration procedures.The analyses were performed by means of powder X-ray diffraction (XRD), micro-Fourier transform infrared (FTIR) and micro-Raman spectroscopies. Calcite, iron oxides, silicates and sodium chloride (from salted holy water) were found in the red coloured calcarenite. On and under the lead coverings, different lead oxides (mainly massicot), other lead salts (lead basic carbonate, cerussite, plumbonacrite Pb10O(OH)6(CO3)6 and lead–chlorine compounds as laurionite PbOHCl and phosgenite Pb2CO3Cl2) were identified by Raman spectroscopy and XRD. Haematite α-Fe2O3, goethite α-FeO(OH) and lepidocrocite γ-FeO(OH) were found on and around the iron hinges.Lead compounds and sodium chloride, through crystallization and solubilization cycles, were responsible for the stone's degradation, whereas the iron corrosion materials on the hinges produced mechanical stress and cracks in the stone.Various suggestions have been given on how to restore these fonts and to remove the causes of damage.
Keywords: Raman spectroscopy; Holy water fonts; Degradation products; X-ray diffraction (XRD); Fourier transform infrared spectroscpy (FTIR);

Two dimensional (2D) homonuclear 1H J-resolved (JRES) nuclear magnetic resonance spectroscopy is increasingly used in metabolomics. This approach visualises metabolite chemical shifts and scalar couplings along different spectral dimensions, thereby increasing peak dispersion and facilitating spectral assignments and accurate quantification. Here, we optimise the processing of 2D JRES spectra by evaluating different window functions, a traditional sine-bell (SINE) and a combined sine-bell–exponential (SEM) function. Furthermore, we evaluate different projection methods for generating 1D projected spectra (pJRES). Spectra were recorded from three disparate types of biological samples and evaluated in terms of sensitivity, reproducibility and resolution. Overall, the SEM window function yielded considerably higher sensitivity and comparable spectral reproducibility and resolution compared to SINE, for both 1D pJRES and 2D JRES datasets. Furthermore, for pJRES spectra, the highest spectral quality was obtained using SEM combined with skyline projection. These improvements lend further support to utilising 2D J-resolved spectroscopy in metabolomics.
Keywords: J-resolved spectroscopy; nuclear magnetic resonance; Metabolomics; Sine-bell window function; Combined sine-bell–exponential window function; Skyline projection; Summation projection;

A thermostat chip of indium-tin oxide glass substrate for static chip polymerase chain reaction (PCR) is, for the first time, introduced in this paper. The transparent conductive layer was used as an electro-heating element. Pulse width modulation and fuzzy proportional integration–differentiation algorithm were adopted in the temperature programming of the chip. The temperature distribution was investigated, and a dynamic control precision within ±2 °C was achieved. The highest ramping rates were 37 °C s−1 for heating and 8 °C s−1 for cooling with an electric fan. The PCR reaction vials were constructed with polyethylene tubes or poly(dimethylsiloxane) directly on the thermostat chip; the chip had a typical size of 25 mm × 25 mm and a thickness of 1.1 mm. Static chip PCR was successfully demonstrated either in a single vial or in an up to 8-parallel array vials. In situ real time fluorescence monitoring during PCR of a λ DNA fragments (236 bp) with SYBR Green I was demonstrated using a blue light emission diode as a light source and a photomultiplier as a detector. The method proposed here is characterized by open access, easy fabrication and low cost. This work could be the basis for developing a portable real time PCR system with disposable chips for point of care tests.
Keywords: Indium-tin oxide glass; Chip thermostat; Static chip polymerase chain reaction; In situ real time fluorescence monitoring; Light emission diode;

Fungal pathogenic nucleic acid detection achieved with a microfluidic microarray device by Lin Wang; Paul C.H. Li; Hua-Zhong Yu; Ash M. Parameswaran (97-104).
Detection of polymerase chain reaction (PCR) products obtained from cultured greenhouse fungal pathogens, Botrytis cinerea and Didymella bryoniae has been achieved using a previously developed microfluidic microarray assembly (MMA) device. The flexible probe construction and rapid DNA detection resulted from the use of centrifugal pumping in the steps of probe introduction and sample delivery, respectively. The line arrays of the oligonucleotide probes were “printed” on a CD-like glass chip using a polydimethylsiloxane (PDMS) polymer plate with radial microfluidic channels, and the sample hybridizations were conducted within the spiral channels on the second plate. The experimental conditions of probe immobilization and sample hybridization were optimized, and both complementary oligonucleotides and PCR products were tested. We were able to achieve adequate fluorescent signals with a sample load as small as 0.5 nM (1 μL) for oligonucleotide samples; for PCR products, we achieved detection at the level of 3 ng.
Keywords: Fungal pathogen; Microfluidic microarray; Centrifugal pumping; Nucleic acid hybridization;

A double-labelled synthetic oligonucleotide is used as a fluorescent molecular aptamer beacon for the reagentless determination of total hardness in tap and bottled waters. Modified thrombin binding aptamer (5′-NH-C3-GGTTGGTGTGGTTGG-C3-SH-3′) carrying 6-carboxyfluorescein (FAM) and 7-amino-4-methyl-coumarin labels at 5′ and 3′, respectively, was used for the simultaneous combined measurement of Mg2+ and Ca2+ cations. Interference from the K+ cation is eliminated via selective tuning of the assay conditions, increasing the temperature beyond the melting point of the potassium-stabilised quadruplex facilitating its liberation from the quadruplex, whilst maintaining the integrity of the magnesium/calcium-stabilised structure. No interference from other cations found in tap or bottled water was observed. The detection limit of the aptamer beacon is 0.04 mmol L−1, with a dynamic linear range of 0–0.5 μM and is very reproducible, with an R.S.D. = 8%, n  = 3. The fluorescent molecular beacon is applied to the determination of total hardness in tap and bottled waters and its’ performance compared to that of the standard method of complexiometric titration and atomic absorption spectroscopy, with an excellent correlation observed. Further work is focused on the immobilization of the aptamer for the development of a re-usable fluorescent/electrochemical aptasensor, for the determination of water hardness.
Keywords: Fluorescent molecular aptamer beacon; Hardness determination; Rapid water analysis;

We present a poly(guanine)-functionalized silica nanoparticle (NP) label-based electrochemical immunoassay for sensitively detecting 2,4,6-trinitrotoluene (TNT). This immunoassay takes advantage of magnetic bead-based platform for competitive displacement immunoreactions and separation, and use electroactive nanoparticles as labels for signal amplification. For this assay, anti-TNT-coated magnetic beads interacted with TNT analog-conjugated poly(guanine)-silica NPs and formed analog-anti-TNT immunocomplexes on magnetic beads. The immunocomplexes coated magnetic beads were exposed to TNT samples, which resulted in displacing the analog conjugated poly(guanine) silica NPs into solution by TNT. In contrast, there are no guanine residues releasing into the solution in the absence of TNT. The reaction solution was then separated from the magnetic beads and transferred to the electrode surface for electrochemical measurements of guanine oxidation with Ru(bpy)3 2+ as mediator. The sensitivity of this TNT assay was greatly enhanced through dual signal amplifications: (1) a large amount of guanine residues on silica nanoparticles are introduced into the test solution by displacement immunoreactions and (2) a Ru(bpy)3 2+-induced guanine catalytic oxidation further enhances the electrochemical signal. Some experimental parameters for the nanoparticle label-based electrochemical immunoassay were studied and the performance of this assay was evaluated. The method is found to be very sensitive and the detection limit of this assay is ∼0.1 ng mL−1 TNT. The electrochemical immunoassay based on the poly[guanine]-functionalized silica NP label offers a new approach for sensitive detection of explosives.
Keywords: Functionalized silica nanoparticle; Electrochemical immunoassay; Poly(guanine); 2,4,6-Trinitrotoluene; Magnetic bead;

A surface plasmon resonance probe without optical fibers as a portable sensing device by Takuo Akimoto; Syunsuke Wada; Isao Karube (119-124).
A surface plasmon resonance (SPR) sensor integrating a small sensor probe, a laser emission diode, a photo detector, and a polarizer was developed as a portable sensing device. The sensor probe was made with a glass cylinder, 50 mm long and 1.5 mm in diameter, that was connected directly to a beam splitter without optical fibers. The SPR spectrum obtained with this probe system showed a 10% reflectivity minimum at 690 nm. Shifts of the SPR spectrum induced by refractive index (RI) changes in the sample were measured by detecting the reflection light intensity at 670 nm. When the sensitivity was compared using a BIAcore™ SPR instrument, the lowest sensor response of 1 mV observed with the SPR probe system coincided with 1.4 × 10−6 of the RI changes. The RI resolution of the SPR probe was estimated with experimentally evaluated noise on the signal, and, consequently, it was concluded that the RI resolution was 1.2 × 10−5. Moreover, immunoreaction was demonstrated with adsorbed bovine serum albumin (BSA) and anti-BSA antibody as an analyte. As a result, 50 ng mL−1 of the lower detection limit was estimated.
Keywords: Surface plasmon resonance; Sensor probe; Biosensor;

Rapid preseparation of interferences for ion mobility spectrometry by Abu B. Kanu; Ching Wu; Herbert H. Hill (125-134).
Two new approaches to reduce false positive interferences commonly observed with explosives and drugs detection in the field were reported for ion mobility spectrometry (IMS). One of the approaches involved the rapid preseparation of potential interferences prior to detection by IMS. Firstly, it was found that the introduction of a short column packed with adsorption packing material before an IMS could help to reduce the false positive rates. Secondly, the retention time at which the most intense response occurred over the analysis time period could be utilized to separate false positive responses from target analytes with the same drift times. Rapid preseparation of potential interferences provided a greater degree of confidence for the detection (in less than 30 s) of drugs, explosives and chemical warfare agents (CWAs). Detection limits as low as 10 pg of TNT with a sensitivity of 12 A g−1 were reported. Successful development of this technique may lead to the construction of a simple interface fitted with a short column of adsorption packing material to enhance either initial separation or to hold-back interferences mixed with explosive and drug responses in the field.
Keywords: Ion mobility spectrometry; Preseparation; Explosives; Drugs; Chemical warfare agents;

The dispersive liquid–liquid microextraction (DLLME) was combined with the flame atomic absorption spectrometry (FAAS) for determination of lead in the water samples. Diethyldithiophosphoric acid (DDTP), carbon tetrachloride and methanol were used as chelating agent, extraction solvent and disperser solvent, respectively. A new FAAS sample introduction system was employed for the microvolume nebulization of the non-flammable chlorinated organic extracts. Injection of 20 μL volumes of the organic extract into an air–acetylene flame provided very sensitive spike-like and reproducible signals.Some effective parameters on the microextraction and the complex formation were selected and optimized. These parameters include extraction and disperser solvent type as well as their volume, extraction time, salt effect, pH and amount of the chelating agent. Under the optimized conditions, the enrichment factor of 450 was obtained from a sample volume of 25.0 mL. The enhancement factor, calculated as the ratio of the slopes of the calibration graphs with and without preconcentration, which was about 1000. The calibration graph was linear in the range of 1–70 μg L−1 with a detection limit of 0.5 μg L−1. The relative standard deviation (R.S.D.) for seven replicate measurements of 5.0 and 50 μg L−1 of lead were 3.8 and 2.0%, respectively. The relative recoveries of lead in tap, well, river and seawater samples at the spiking level of 20 μg L−1 ranged from 93.8 to 106.2%. The characteristics of the proposed method were compared with those of the liquid–liquid extraction (LLE), cloud point extraction (CPE), on-line and off-line solid-phase extraction (SPE) as well as co-precipitation, based on bibliographic data. Operation simplicity, rapidity, low cost, high enrichment factor, good repeatability, and low consumption of the extraction solvent at a microliter level are the main advantages of the proposed method.
Keywords: Dispersive liquid–liquid microextraction; Preconcentration; Flame atomic absorption spectrometry; Microsample introduction; Lead; Water analysis;

Facile synthesis and functionalization of water-soluble gold nanoparticles for a bioprobe by Nishima Wangoo; K.K. Bhasin; Robin Boro; C. Raman Suri (142-148).
In this work, we report the size tunable synthesis of water-dispersed gold nanoparticles by using octadecylamine (ODA) as the reducing agent, that electrostatically complexes with the chloroaurate ions, reduces them, and subsequently caps the gold nanoparticles. Amine-capped gold nanoparticles, thus formed, were subsequently coordinated with a secondary monolayer of an anionic surfactant, sodium bis(2-ethylhexyl)-sulfosuccinate (AOT) which helps in providing sufficient hydrophilicity to the gold nanoparticles. Functionalized gold nanoparticles were characterized by UV–vis, IR spectrophotometric, dynamic light scattering, zeta-potential and transmission electron microscopic techniques, which demonstrated high stability of gold nanoparticles in aqueous media, indicating stabilization via bilayers of ODA and AOT. The gold nanoparticles were further conjugated with a protein (bovine serum albumin) and the interaction was investigated by circular dichroism studies as well as by measuring the fluorescence quenching of the tryptophan residues of protein molecules after the binding of nanoparticles to specific sites of the protein. The binding constant and the stoichiometry values indicated that the particles with larger core size are less site-specific but show higher binding affinity with protein molecules. The use of a bio-compatible synthetic process and the stabilization of the gold nanoparticles by ODA and AOT are interesting from the point of view of making bioprobes for life science applications.
Keywords: Amine-capped gold nanoparticles; Octadecylamine; Bioconjugation; Binding constants;