This study describes the synthesis, characterization and application of a new chrysin-based silica core-shell magnetic nanoparticles (Fe3O4@SiO2-N-chrysin) as an adsorbent for the preconcentration of Cu(II) from aqueous environment. The morphology, thermal stability and magnetic property of Fe3O4@SiO2-N-chrysin were analyzed using FTIR, FESEM, TEM, XRD, thermal analysis and VSM. The extraction efficiency of Fe3O4@SiO2-N-chrysin was analyzed using the batch wise method with flame atomic absorption spectrometry. Parameters such as the pH, the sample volume, the adsorption-desorption time, the concentration of the desorption solvent, the desorption volume, the interference effects and the regeneration of the adsorbent were optimized. It was determined that Cu(II) adsorption is highly pH-dependent, and a high recovery (98%) was achieved at a pH 6. The limit of detection (S/N=3), the limit of quantification (S/N=10), the preconcentration factor and the relative standard deviation for Cu(II) extraction were 0.3 ng mL(-1), 1 ng mL(-1), 100 and 1.9% (concentration=30 ng mL(-1), n=7), respectively. Excellent relative recoveries of 97-104% (%RSD<3.12) were achieved from samples from a spiked river, a lake and tap water. The MSPE method was also validated using certified reference materials SLRS-5 with good recovery (92.53%).
The development of an optical biosensor based on immobilization of 3-methyl-2-benzothiazolinone hydrazone (MBTH) in hybrid nafion/sol-gel silicate film and tyrosinase in chitosan film for the detection of phenolic compounds has been described. Tyrosinase was immobilized in chitosan film deposited on the hybrid nafion/sol-gel silicate film containing MBTH. The enzymatic oxidation product of phenolic compounds were stabilized through formation of adduct with MBTH to produce a maroon color adduct. The color intensity of adduct was found to increase proportionally with the increase of the substrate concentrations after 5min exposure. The linearity of the biosensor towards phenol, catechol and m-cresol were in the respective concentration range of 0.5-7.0, 0.5-10.0 and 1.0-13.0mg/L with detection limit of 0.18, 0.23 and 0.43mg/L, respectively. The biosensor shows a good stability for at least 3 months.
A fast and simple solvent microextraction technique using salting out-vortex-assisted liquid-liquid microextraction (salting out-VALLME) was developed for the extraction of furfurals (2-furfural (2-F), 3-furfural (3-F), 5-methylfurfural (5-MF) and 5-hydroxymethylfurfural (5-HMF)) and patulin (PAT) in fruit juice samples. The optimum extraction conditions for 5 mL sample were: extraction solvent, 1-hexanol; volume of extractant, 200 µL; vortex time, 45 s; salt addition, 20%. The simultaneous determination of the furfurals and PAT were investigated using high performance liquid chromatography coupled with diode array detector (HPLC-DAD). The separation was performed using ODS Hypersil C18 column (4.6 mm i.d × 250 mm, 5 μm) under gradient elution. The detection wavelengths used for all compounds were 280 nm except for 3-F (210 nm). The furfurals and PAT were successfully separated in less than 9 min. Good linearities (r(2)>0.99) were obtained within the range 1-5000 μg L(-1) for all compounds except for 3-F (10-5000 µg L(-1)) and PAT (0.5-100 μg L(-1)). The limits of detection (0.28-3.2 µg L(-1)) were estimated at S/N ratio of 3. The validated salting out-VALLME-HPLC method was applied for the analysis of furfurals and PAT in fruit juice samples (apple, mango and grape).
An optical sensor for Hg(II) monitoring using a complex of zinc dithizonate immobilised on XAD 7 which is based on reflectance spectrophotometry has been developed in this study. Measurements were made using a kinetic approach whereby the reflectance signal is measured at a fixed time of 5 min. The sensor could be regenerated using a saturated solution of KCl in 1 M sulphuric acid. The sensor was found to have an optimum response at pH 3.0 with respective measurement repeatability and probe-to-probe reproducibility of 1.53% and 5.26%. A linear response was observed in the Hg(II) concentration range of 0.0-180.0 ppm with a calculated limit of detection (LOD) of 0.05 ppm. The results obtained for aqueous Hg(II) determination using this probe were found to be comparable with the well-established method of atomic absorption spectrometry.
Chrome azurol S immobilised on XAD-2 has been used in this study as a reagent phase for the development of an optical fibre Al(III) sensor. Using a kinetic approach, this sensor was able to give a linear response in the Al(III) concentration range of 1.3 x 10(-5)-2.0 x 10(-4) M with a limit of detection of 1.0 x 10(-4) M. The optimum responses were obtained at pH 6.0 and when the solution was stirred. The sensor response was found to have a repeatability and reproducibility of 1.6% and 5.8%, respectively. The results obtained for Al(III) determination in aqueous sample were in good agreement with those obtained using graphite furnace-atomic absorption spectrometry.
A new sample preparation method, ion-pair vortex assisted liquid-liquid microextraction (VALLME-BE), for the determination of a highly polar anti-diabetic drug (metformin) in plasma sample was developed. The VALLME-BE was performed by diluting the plasma in borate buffer and extracted to 150µL 1-octanol containing 0.2M di-(2-ethylhexyl)phosphoric acid as intermediate phase. The drug was next back-extracted into 20µL of 0.075M HCl solution. The effects of pH, ion-pair concentration, type of organic solvent, volume of extraction phases, ionic strength, vortexing and centrifugation times on the extraction efficiency were investigated. The optimum conditions were at pH 9.3, 60s vortexing and 2min centrifugation. The microextract, contained metformin and buformin (internal standard), was directly injected into a HPLC unit using C1 column (250mm×4.6mm×10µm) and detected at 235nm. The method was validated and calibration curve was linear with r2>0.99 over the range of 20-2000µgL-1. The limits of detection and quantitation were 1.4 and 4.1µgL-1, respectively. The accuracy was within 94.8-108% of the nominal concentration. The relative standard deviation for inter- and intra-day precision was less than 10.8%. The method was conveniently applied for the determination of metformin in plasma samples.
A new lead(II) electrode has been constructed with poly(hydroxamic acid) (PHXA) as the active material and silicone rubber as the supporting material. The electrode gave near Nerstian response over the concentration range 4 x 10(-5)-1 x 10(-2)M lead(II). The detection limit of the electrode is approximately 4 x 10(-6)M and the electrode works well in the pH range 4.5-6.0. The response time was 50-120 sec over the whole concentration range and the electrode has a working life of at least 4 weeks. Iron(III) severely poisoned the electrode membrane. Nickel(II) and mercury(II) gave very strong interference compared to copper(II), silver(I), cobalt(II), sodium(I), potassium(I), zinc(II) and cadmium(II) which gave some or little interference. Values determined with atomic absorption (AAS) and a commercial lead(II) electrode were in good agreement with those measured with the lead(II) electrode reported here.
The characteristics of a potentiometric biosensor for the determination of permethrin in treated wood based on immobilised cells of the fungus Lentinus sajor-caju on a potentiometric transducer are reported this paper. The potentiometric biosensor was prepared by immobilisation of the fungus in alginate gel deposited on a pH-sensitive transducer employing a photocurable acrylic matrix. The biosensor gave a good response in detecting permethrin over the range of 1.0-100.0 µM. The slope of the calibration curve was 56.10 mV/decade with detection limit of 1.00 µM. The relative standard deviation for the sensor reproducibility was 4.86%. The response time of the sensor was 5 min at optimum pH 8.0 with 1.00 mg/electrode of fungus L. sajor-caju. The permethrin biosensor performance was compared with the conventional method for permethrin analysis using high performance liquid chromatography (HPLC), and the analytical results agreed well with the HPLC method (at 95% confidence limit). There was no interference from commonly used organophosphorus pesticides such as diazinon, parathion, paraoxon, and methyl parathion.
This research work demonstrates compositional engineering of an organic-inorganic hybrid nano-composites for modifying absolute threshold of humidity sensors. Vanadyl-2,9,16,23-tetraphenoxy-29H,31H-phthalocyanine (VOPcPhO), an organic semiconductor, doped with Titanium-dioxide nanoparticles (TiO2NPs) has been employed to fabricate humidity sensors. The morphology of the VOPcPhO:TiO2nano-composite films has been analyzed by atomic force microscopy (AFM) and field emission scanning electron microscopy (FESEM). The sensors have been examined over a wide range of relative humidity i.e. 20-99% RH. The sensor with TiO2(90nm) shows reduced sensitivity-threshold and improved linearity. The VOPcPhO:TiO2(90nm) nano-composite film is comprised of uniformly distributed voids which makes the surface more favorable for adsorption of moisture content from environment. The VOPcPhO:TiO2nano-composite based sensor demonstrates remarkable improvement in the sensing parameter when equated with VOPcPhO sensors.
The aim of this study was to optimize the antioxidant activity of Piper nigrum L. essential oil extracted using the supercritical carbon dioxide (SC-CO₂) technique. Response surface methodology was applied using a three-factor central composite design to evaluate the effects of three independent extraction variables: pressure of 15-30 MPa, temperature of 40-50 °C and dynamic extraction time of 40-80 min. The DPPH radical scavenging method was used to evaluate the antioxidant activity of the extracts. The results showed that the best antioxidant activity was achieved at 30 MPa, 40 °C and 40 min. The extracts were analyzed by GC-FID and GC-MS. The main components extracted using SC-CO₂ extraction in optimum conditions were β-caryophyllene (25.38 ± 0.62%), limonene (15.64 ± 0.15%), sabinene (13.63 ± 0.21%), 3-carene (9.34 ± 0.04%), β-pinene (7.27 ± 0.05%), and α-pinene (4.25 ± 0.06%). The essential oil obtained through this technique was compared with the essential oil obtained using hydro-distillation. For the essential oil obtained by hydro-distillation, the most abundant compounds were β-caryophyllene (18.64 ± 0.84%), limonene (14.95 ± 0.13%), sabinene (13.19 ± 0.17%), 3-carene (8.56 ± 0.11%), β-pinene (9.71 ± 0.12%), and α-pinene (7.96 ± 0.14%). Radical scavenging activity of the extracts obtained by SC-CO₂ and hydro-distillation showed an EC₅₀ of 103.28 and 316.27 µg mL(-1) respectively.
A new analytical method for the simultaneous determination of the antidiabetic drugs rosiglitazone (ROS) and metformin hydrochloride (MH) with marked differences in their affinity towards organic solvents (log P of 2.4 and -1.43, respectively) was developed. Prior to the HPLC separation, the drugs were subjected to a sequential hollow fiber liquid phase microextraction (HF-LPME) procedure. Two sequential HF-LPME approaches were considered, the preferred one involves the use of two vials containing solution mixtures for the extraction of ROS (vial 1) and MH (vial 2), respectively, but using the same fiber and acceptor phase. Important parameters that affect the extraction efficiency such as extracting solvent, donor phase conditions, HCl concentration, agitation, extraction time, addition of salt, etc. were studied. Under the optimum conditions, good enrichment factors (EF, 471 and 86.6 for ROS and MH, respectively) were achieved. Calibration curves were linear over the range 1-500 (r(2)=0.998) and 5-2500 ng mL(-1) (r(2)=0.999) for ROS and MH, respectively. The relative standard deviation values (RSD%) for six replicates were below 8.4%. Detection and quantitation limits based on S/N ratio of 3 and 10 were 0.12, 1.0 and 0.36, 3.0 ng mL(-1) for ROS and MH, respectively. The proposed method is simple, sensitive and opens up new opportunities for the microextraction of analytes with contrasting properties.
A flow injection (FI) method with on-line preconcentration using a mini-column loaded with 8-hydroxyquinoline immobilized on controlled pore glass (CPG-8HQ) is described for the determination of trace metals by ion chromatography (IC) with pyridine-2-6-dicarboxylic acid (PDCA) as the eluent. Copper, cadmium, lead, zinc, nickel and iron were determined at ppb level after post-column derivatization with 4-(2-pyridylazo)-resorcinol (PAR). The detection limits (3sigma) for the FI/IC system were 8.27, 0.89, 0.09, 0.06, 0.09 and 0.07 g l(-1) for Pb(2+), Cd(2+), Cu(2+) Ni(2+), Zn(2+) and Fe(3+), respectively, using 5 ml sample volume. The method was applied to the analysis of Malaysian natural waters.
A simple atomic-absorption spectrophotometry method is described for the determination of silver, bismuth, cadmium, copper, iron, nickel and zinc in lead- and tin-base solders and white-metal bearing alloys, with use of a single sample solution. The sample is dissolved in a mixture of hydrobromic acid and bromine, then fumed with sulphuric acid. The lead sulphate is dissolved in concentrated hydrochloric acid. The method is particularly suitable for the determination of silver and bismuth, which are co-precipitated with lead sulphate. The other elements can also be determined after removal of the lead sulphate by filtration.
Electro-mediated microextraction (EMM) combined with micro-high performance liquid chromatography-ultraviolet detection was successfully developed for the determination of selected phenols, namely 4-chlorophenol (4CP), 2-nitrophenol (2NP) and 2,4-dichlorophenols (2,4 DCP) in water. A solvent-impregnated agarose gel disc was utilized as a solvent holder in this study. Under optimum extraction conditions, the method showed good linearity in the range of 0.1-250µgL-1, 0.3-250µgL-1and 0.2-500µgL-1for 4CP, 2NP and 2,4 DCP, respectively with correlation coefficients of ≥ 0.9975, ultra-trace LODs (0.03-0.1µgL-1) and satisfactory relative recovery average (85.0-114.1%) for the analysis of selected phenols. The proposed method was rapid and eco-friendly as the solvent holder was constructed using minute amounts of extraction solvent immobilized within the biodegradable agarose gel disc. A comparative microextraction technique termed solvent-impregnated agarose gel liquid phase microextraction (AG-LPME) was re-optimized and validated for the extraction of phenols in water. The method offered good linearity, ultra-trace LODs ranging 0.1-0.5µgL-1and satisfactory average of relative recovery (86.1-114.1%). The EMM was superior in terms of sensitivity and time-effectiveness compared to AG-LPME. Both techniques combine extraction and pre-concentration in mini-scaled approaches using an eco-friendly solvent holder that fulfil the green chemistry concept.
Label-free-based detection is pivotal for real-time monitoring of biomolecular interactions and to eliminate the need for labeling with tags that can occupy important binding sites of biomolecules. One simplest form of label-free-based detection is ultraviolet-visible-near-infrared (UV-vis-NIR) spectroscopy, which measure changes in reflectivity as a means to monitor immobilization and interaction of biomolecules with their corresponding partners. In biosensor development, the platform used for the biomolecular interaction should be suitable for different molecular recognition elements. In this study, gold (Au)-coated polycarbonate was used as a platform and as a proof-of-concept, erythropoietin (EPO), a doping substance widely abused by the athletes was used as the target. The interaction of EPO with its corresponding molecular recognition elements (anti-EPO monoclonal antibody and anti-EPO DNA aptamer) is monitored by UV-vis-NIR spectroscopy. Prior to this, to show that UV-vis-NIR spectroscopy is a suitable method for measuring biomolecular interaction, the interaction between biotin and streptavidin was demonstrated via this strategy and reflectivity of this interaction decreased by 25%. Subsequent to this, interaction of the EPO with anti-EPO monoclonal antibody and anti-EPO DNA aptamer resulted in the decrease of reflectivity by 5% and 10%, respectively. The results indicated that Au-coated polycarbonate could be an ideal biosensor platform for monitoring biomolecular interactions using UV-vis-NIR spectroscopy. A smaller version of the Au-coated polycarbonate substrates can be derived from the recent set-up, to be applied towards detecting EPO abuse among atheletes.
Aptamers are nucleic acid-based molecular recognition elements that are specific and have high binding affinity against their respective targets. On account of their target recognition capacity, aptamers are widely utilized in a number of applications including diagnostics. This review aims to highlight the recent developments of aptasensors expedient for point-of-care (POC) diagnostics. Significant focus is given on the primary assay formats of aptamers such as fluorescence, electrochemical, surface plasmon resonance (SPR) and colorimetric assays. A potpourri of platforms such as paper-based device, lateral flow assay, portable electrodes, portable SPR and smart phones expedient for point-of-care (POC) diagnostics are discussed. Emphasis is also given on the technicalities and assay configurations associated with the sensors.
Dot blot assays have always been associated with antibodies as the main molecular recognition element, which are widely employed in a myriad of diagnostic applications. With the rising of aptamers as the equivalent molecular recognition elements of antibodies, dot blot assays are also one of the diagnostic avenues that should be scrutinized for their amenability with aptamers as the potential surrogates of antibodies. In this review, the stepwise procedures of an aptamer-based dot blot assays are underscored before reviewing the existing aptamer-based dot blot assays developed so far. Most of the applications center on monitoring the progress of SELEX and as the validatory assays to assess the potency of aptamer candidates. For the purpose of diagnostics, the current effort is still languid and as such possible suggestions to galvanize the move to spur the aptamer-based dot blot assays to a point-of-care arena are discussed.
A capillary electrophoretic method for the separation of the aminoglutethimide (AGT) enantiomers using methylated-beta-cyclodextrin (M-beta-CD) as chiral selector is described. Several parameters affecting the separation were studied, including the type and concentration of chiral selector, buffer pH, voltage and temperature. Good chiral separation of the racemic mixture was achieved in less than 9 min with resolution factor Rs=2.1, using a fused-silica capillary and a background electrolyte (BGE) of tris-phosphate buffer solution (50 mmol L(-1), pH 3.0) containing 30 mgm L(-1) of M-beta-CD. The separation was carried out in normal polarity mode at 25 degrees C, 16 kV and using hydrostatic injection. Acceptable validation criteria for selectivity, linearity, precision, and accuracy/recovery were included. The proposed method was successfully applied to the assay of AGT enantiomers in pharmaceutical formulations. The computational calculations for the inclusion complexes of the R- and S-AGT-M-beta-CD rationalized the reasons for the different migration times between the AGT enantiomers.
The developments in mass spectrometry (MS) in the past few decades reveal the power and versatility of this technology. MS methods are utilized in routine analyses as well as research activities involving a broad range of analytes (elements and molecules) and countless matrices. However, manual MS analysis is gradually becoming a thing of the past. In this article, the available MS automation strategies are critically evaluated. Automation of analytical workflows culminating with MS detection encompasses involvement of automated operations in any of the steps related to sample handling/treatment before MS detection, sample introduction, MS data acquisition, and MS data processing. Automated MS workflows help to overcome the intrinsic limitations of MS methodology regarding reproducibility, throughput, and the expertise required to operate MS instruments. Such workflows often comprise automated off-line and on-line steps such as sampling, extraction, derivatization, and separation. The most common instrumental tools include autosamplers, multi-axis robots, flow injection systems, and lab-on-a-chip. Prototyping customized automated MS systems is a way to introduce non-standard automated features to MS workflows. The review highlights the enabling role of automated MS procedures in various sectors of academic research and industry. Examples include applications of automated MS workflows in bioscience, environmental studies, and exploration of the outer space.
In this work, we proposed a biosensor for trypsin proteolytic activity assay using immobilization of model peptides on screen-printed electrodes (SPE) modified with gold nanoparticles (AuNPs) prepared by electrosynthetic method. Sensing of proteolytic activity was based on electrochemical oxidation of tyrosine residues of peptides. We designed peptides containing N-terminal cysteine residue for immobilization on an SPE, modified with gold nanoparticles, trypsin-specific cleavage site and tyrosine residue as a redox label. The peptides were immobilized on SPE by formation of chemical bonds between mercapto groups of the N-terminal cysteine residues and AuNPs. After the incubation with trypsin, time-dependent cleavage of the immobilized peptides was observed by decline in tyrosine electrochemical oxidation signal. The kinetic parameters of trypsin, such as the catalytic constant (kcat), the Michaelis constant (KM) and the catalytic efficiency (kcat/KM), toward the CGGGRYR peptide were determined as 0.33 ± 0.01 min-1, 198 ± 24 nM and 0.0016 min-1 nM-1, respectively. Using the developed biosensor, we demonstrated the possibility of analysis of trypsin specificity toward the peptides with amino acid residues disrupting proteolysis. Further, we designed the peptides with proline or glutamic acid residues after the cleavage site (CGGRPYR and CGGREYR), and trypsin had reduced activity toward both of them according to the existing knowledge of the enzyme specificity. The developed biosensor system allows one to perform a comparative analysis of the protease steady-state kinetic parameters and specificity toward model peptides with different amino acid sequences.