Methylmercury determination using a hyphenated high performance liquid chromatography ultraviolet cold vapor multipath atomic absorption spectrometry system

The present work investigates the use of a multipath cell atomic absorption mercury detector for mercury speciation analysis in a hyphenated high performance liquid chromatography assembly. The multipath absorption cell multiplies the optical path while energy losses are compensated by a very intense primary source. Zeeman-effect background correction compensates for non-specific absorption. For the separation step, the mobile phase consisted in a 0.010% m/v mercaptoethanol solution in 5% methanol (pH = 5), a C₁₈ column was used as stationary phase, and post column treatment was performed by UV irradiation (60 °C, 13 W). The eluate was then merged with 3 mol L^− 1 HCl, reduction was performed by a NaBH₄ solution, and the Hg vapor formed was separated at the gas–liquid separator and carried through a desiccant membrane to the detector. The detector was easily attached to the system, since an external gas flow to the gas–liquid separator was provided. A multivariate approach was used to optimize the procedure and peak area was used for measurement. Instrumental limits of detection of 0.05 µg L^− 1 were obtained for ionic (Hg²⁺) and HgCH₃⁺, for an injection volume of 200 µL. The multipath atomic absorption spectrometer proved to be a competitive mercury detector in hyphenated systems in relation to the most commonly used atomic fluorescence and inductively coupled plasma mass spectrometric detectors. Preliminary application studies were performed for the determination of methyl mercury in sediments.     

Portable flow-injection analyzer with liquid-core waveguide based fluorescence, luminescence, and long path length absorbance detector

We describe a multifunctional flow analysis instrument that is portable ( cm, 2.3 kg) for facile field deployment. Using a 50 cm long Teflon^® AF tubing as final reaction and optical measurement conduit, we combine a liquid-core waveguide (LCW) based fluorescence detector that is transversely illuminated by an addressable light emitting diode array, a chemiluminescence (CL) detector and an absorbance detector with a solid-state broadband (400-700 nm) source. Several illustrative experiments have been carried out to test the performance of the instrument in different detection modes. A S/N=3 limit of detection (LOD) of 0.25 μg l⁻¹ for chromium(VI) was established using the diphenylcarbazide chemistry, and an LOD of 5 μg l⁻¹ was similarly established for Al(III), using Pyrocatechol Violet (PCV) as the chelating chromogenic dye, in both cases using long path absorption detection. The LOD for aqueous hydrogen peroxide was 16 nM using a fluorescence method based on the formation of thiochrome from thiamine and 4 nM by a luminol chemiluminescence method. With a Nafion membrane diffusion scrubber (DS), the LOD was 8.0 pptv for gaseous hydrogen peroxide by the fluorescence method.     

A novel solid-state electrochemiluminescence detector for capillary electrophoresis based on tris(2,2'-bipyridyl)ruthenium(II) immobilized in Nafion/PTC-NH2 composite film

A new electrochemiluminescence (ECL) detector for capillary electrophoresis (CE) based on tris(2,2′-bipyridyl)ruthenium(II) (Ru(bpy)₃²⁺) immobilized in Nafion/PTC-NH₂ (an ammonolysis product of 3,4,9,10-perylenetetracarboxylic dianhydride (PTCDA)) composite film was presented for the first time. The Nafion/PTC-NH₂ composite film could effectively immobilize tris(2,2′-bipyridyl)ruthenium(II) via ion-exchange and electrostatic interaction. Cyclic voltammetric and ECL behavior of Nafion/PTC-NH₂/Ru composite film was investigated compared to Nafion/Ru composite. The Nafion/PTC-NH₂/Ru composite film exhibited good ECL stability and simple operability. Then the CE with solid-state ECL detector system was successfully used to detect sophora - a quinolizidine type - alkaloids as sophoridine (SR) and matrine (MT). The CE-ECL parameters that affected separation and detection were optimized. Under the optimized conditions, the linear range was from 2.5 × 10⁻⁸ to 2 × 10⁻⁶ mol/L for SR, 1.0 × 10⁻⁸ to 1.0 × 10⁻⁶ mol/L for MT. The detection limit (S/N = 3) was estimated to be 5 × 10⁻⁹ and 10⁻⁹ mol/L for SR and MT, respectively. It was shown that the CE coupling with solid-state ECL detector system exhibited satisfying sensitivity of analysis.     

Cavity Ring-Down Spectroscopy coupled with helium microwave-induced plasma for fluorine detection

Cavity Ring-Down Spectroscopy with a tunable diode laser has been used for the detection of fluorine at trace levels. A homemade experimental setup was constructed to accommodate the microwave-induced plasma glow discharge inside the optical cavity. The energy delivered by helium plasma discharge was successfully used not only for the dissociation of fluorine molecules but, more importantly, also for the excitation of fluorine atoms into their metastable level 3s⁴P_5/2. The absorption observed at the 3s⁴P_5/2–3p⁴D^o_7/2 transition (corresponding to a 685.603-nm atomic line) was used for fluorine atom detection. Good agreement between theoretical and measured absorption line shapes confirmed the selectivity of fluorine atomic absorption. We observed an absorption coefficient of 4 ? 10⁻⁶ cm⁻¹, corresponding to a detection of F(3s⁴P_5/2) atoms at about 1.5 ? 10⁶ atoms cm⁻³. These data demonstrate that we achieved a detection limit of 100 ng g⁻¹ under the conditions of our initial homemade experimental setup. However, the optimization of the system may lead to the improvement of the detection limit by approximately two orders of magnitude.     

Monitoring inorganic mercury and methylmercury species with liquid chromatography-piezoelectric detection

A piezoelectric detection system coupled with a liquid chromatographic system is developed for the speciation of inorganic mercury(II) and methylmercury. Piezoelectric detection has been demonstrated to be a very sensitive detection system for total mercury determination when a gold-coated piezoelectric quartz crystal was used. The analytical features of this detection unit make it very suitable to be used as a detector coupled with a liquid chromatographic system for monitoring mercury species. After separation by a chromatography column (Spherisorb ODS-2, 5 μm,  mm i.d.), mercury species are liberated and reduced, by using stannous chloride, and are detected as an amalgam on the gold-coated piezoelectric quartz crystal, the sensor subsequently was regenerated by passing a peroxydisulfate solution. This detector exhibits good sensitivity, it allows the determination of mercury at sub-ppb concentration levels (0.30-1.20 μg l⁻¹). The precision, expressed as relative standard deviation, was ±2.7% (n=11) for a 0.5 μg l⁻¹ total mercury concentration. The proposed method is free of interferences and it allowed the determination of inorganic mercury and methylmercury species in natural waters.     

Macroporous ordered titanium dioxide (TiO2) inverse opal as a new label-free immunosensor

Photonic crystal sensing materials have been validated that they are very sensitive to refractive index changes. Herein, three-dimensionally ordered macroporous (3DOM) (>50 nm) TiO₂ inverse opal film has been fabricated by the self-assembly technique. Based on the TiO₂ inverse opal film, the optical spectrometer was established for label-free immunosensor. The sensing performance of the 3DOM TiO₂ was investigated using human IgG/goat anti-human IgG couple, which showed that the sensitivity of 3DOM TiO₂ inverse opal film could reach to 1 μg mL⁻¹ (equivalent to 1.5 pg mm⁻²) of protein concentration detection limit. The 3DOM TiO₂ inverse opal has a large internal surface area, low fluorescence background and unique optical properties. These characteristics indicated the feasibility of 3DOM TiO₂ inverse opal in label-free immunoassay.     

A gravity driven micro flow injection wetting film extraction system on a polycarbonate chip

A micro flow injection wetting film liquid-liquid extraction system has been developed for trace analyte concentration and on-chip detection. A hydrophobic channel fabricated on a polycarbonate chip was used to support the wetting film, and hydrostatic pressure generated by the difference in liquid levels was employed to drive the fluids. Sequential injection of segments of aqueous sample solution and organic solvent was conducted by switching the sample- or solvent-containing vials to an on-chip sampling probe, and detection was performed by a co-focused, laser induced fluorescence detector. Using butyl rhodamine B as a model analyte and butanol as the solvent for both film-coating and elution, various experimental conditions such as hydrostatic pressure, coating time, channel length, sampling volume, and sample acidity were investigated. Under optimized conditions, a 24-fold enrichment factor was obtained with the consumption of about 3 μL sample solution, and a detection limit (3σ) of 6.0 × 10⁻⁹ M butyl rhodamine B was achieved at the sampling rate of 19 h⁻¹. Eleven consecutive runs of a 1.0 × 10⁻⁵ M butyl rhodamine B solution produced a relative standard deviation of 1.5% for the detected fluorescence signals.     

Dual fluorescence/contactless conductivity detection for microfluidic chip

A new dual detection system for microchip is reported. Both fluorescence detector (FD) and contactless conductivity detector (CCD) were combined together and integrated on a microfluidic chip. They shared a common detection position and responded simultaneously. A blue light-emitting diode was used as excitation source and a small planar photodiode was used to collect the emitted fluorescence in fluorescence detection, which made the device more compact and portable. The coupling of the fluorescence and contactless conductivity modes at the same position of a single separation channel enhanced the detection characterization of sample and offered simultaneous detection information of both fluorescent and charged specimen. The detection conditions of the system were optimized. K⁺, Na⁺, fluorescein sodium, fluorescein isothiocyanate (FITC) and FITC-labeled amino acids were used to evaluate the performance of the dual detection system. The limits of detection (LOD) of FD for fluorescein Na⁺, FITC, FITC-labeled arginine (Arg), glycine (Gly) and phenylalanine (Phe) were 0.02 μmol L⁻¹, 0.05 μmol L⁻¹, 0.16 μmol L⁻¹, 0.15 μmol L⁻¹, 0.12 μmol L⁻¹ respectively, and the limits of detection (LOD) of CCD achieved 0.58 μmol L⁻¹ and 0.39 μmol L⁻¹ for K⁺ and Na⁺ respectively.     

Low pressure ion chromatography with a low cost paired emitter-detector diode based detector for the determination of alkaline earth metals in water samples

The use of a low pressure ion chromatograph based upon short (25 mm × 4.6 mm) surfactant coated monolithic columns and a low cost paired emitter-detector diode (PEDD) based detector, for the determination of alkaline earth metals in aqueous matrices is presented. The system was applied to the separation of magnesium, calcium, strontium and barium in less than 7 min using a 0.15 M KCl mobile phase at pH 3, with post-column reaction detection at 570 nm using o-cresolphthalein complexone. A comparison of the performance of the PEDD detector with a standard laboratory absorbance detector is shown, with limits of detection for magnesium and calcium using the low cost PEDD detector equal to 0.16 and 0.23 mg L⁻¹, respectively. Finally, the developed system was used for the determination of calcium and magnesium in a commercial spring water sample.     

A thin-layer contactless conductivity cell for detection in flowing liquids

The design is described of a thin-layer contactless conductivity detector suitable for liquid chromatography and flow-injection analysis. Its principal analytical parameters have been determined using a potassium chloride solution: the linear dynamic range extends from 7.5 × 10⁻⁶ to 1.5 × 10⁻² S m⁻¹, corresponding to the KCl concentration range from 0.5 to 1000 μM, the limit of detection equals 3.5 × 10⁻⁶ S m⁻¹ (0.2 μM KCl), the detection repeatability, expressed in terms of the relative standard deviation, amounts to 1.13% and the detection volume is 0.6 μL. The detector was applied to detection of ionic compounds, benzoic, lactic and octanesulfonic acids, and sodium capronate, after their separation by liquid chromatography in a Biospher PSI 100C 18 columns using a 60% aqueous acetonitrile mobile phase. The frequency characteristics of the detector are reasonably theoretically described on the basis of a simple model which is commonly used in the field of contactless impedance detectors.     

Fast on-line ultrasound-assisted extraction coupled to a flow injection-atomic absorption spectrometric system for zinc determination in meat samples

A continuous ultrasound-assisted extraction system connected to a flow injection manifold has been used for the on-line determination of zinc in meat samples by flame atomic absorption spectrometry. An experimental design was used for the optimisation of the continuous manifold. This flow injection methodology allowed a sampling frequency of ca. 80 samples per hour with a relative standard deviation for the whole procedure of 0.3% (for a sample containing 163.6 μg g⁻¹ Zn). The detection limit was 0.6 μg g⁻¹ for a sample amount of 5 mg. Accurate results were obtained by measuring certified reference materials (BCR-186 (pig kidney) and BCR-184 (bovine muscle)). The analytical procedure was applied to different real meat samples with satisfactory results.     

Optical fiber biosensor coupled to chromatographic separation for screening of dopamine, norepinephrine and epinephrine in human urine and plasma

An optical fiber biosensor has been developed for the determination of catecholamines (dopamine, norepinephrine and epinephrine) based on the recognition capacity of the enzyme laccase. In this study, a glass tube constituted by a fused silica fiber coated with a film of polystyrene/divinylbenzene resin (PS/DVB) was used for catecholamines separation. Firstly, the analyzer was tested for calibration and its analytical performance for catecholamines detection was compared with a classical analytical method, namely high performance liquid chromatography-electrochemical detector (HPLC-ED). The developed analytical device shows a high potential for catecholamines quantification with a detection limit of 2.1, 2.6 and 3.4 pg mL⁻¹ for dopamine, norepinephrine and epinephrine, respectively. The analytical sensitivity, inferred from the slope of the calibration curves established for a range of concentrations between 5 and 125 pg mL⁻¹, was found to be 0.344, 0.252 and 0.140 dB/pg mL⁻¹ for dopamine, norepinephrine and epinephrine, respectively. Furthermore, catecholamines speciation with the PS/DVB fiber was completely achieved in 3 min. The analytical performance of the reported sensor was also evaluated and found adequate for catecholamines determination in human urine and plasma samples.     

Quantitative Moisture Measurement with a Cavity Ring-down Spectrometer using Telecom Diode Lasers

Moisture measurement is of great needs in semiconductor industry, combustion diagnosis, meteorology, and atmospheric studies. We present an optical hygrometer based on cavity ring-down spectroscopy (CRDS). By using different absorption lines of H₂O in the 1.56 and 1.36 μm regions, we are able to determine the relative concentration (mole fraction) of water vapor from a few percent down to the 10^-12 level. The quantitative accuracy is examined by comparing the CRDS hygrometer with a commercial chilled-mirror dew-point meter. The high sensitivity of the CRDS instrument allows a water detection limit of 8 pptv.     

Frequency-selective absorbance detection: Refractive index and turbidity compensation with dual-wavelength measurement

A frequency-selective absorbance detection approach and its applications are described. First, a digital signal processor-lock-in amplifier (DSP-LIA)-based absorbance detector was evaluated. Compared to a simple operational amplifier (TL082CP)-based detector, the DSP-LIA-based detector showed lower noise levels, but the relative advantage was reduced under very low photocurrent levels (down to few nA). A 7 cm pathlength flow cell with this commercial LIA-based detector exhibited excellent Beer's law linearity (r² = 0.9999) and a noise level of 7 micro absorbance units (μAU). The limit of detection (LOD, S/N = 3) for methyl orange (MO) was 7 nM with this detector. Finally, as a more affordable alternative to an LIA, a balanced demodulator integrated circuit chip was used to fabricate a dual wavelength-frequency-selective LED-based absorbance detector. This device successfully compensated refractive index (RI) effect and turbidity effect in test flow systems. The LOD for MO with this system was 8 nM.     

An improved biosensor for acetaldehyde determination using a bienzymatic strategy at poly(neutral red) modified carbon film electrodes

Improved biosensors for acetaldehyde determination have been developed using a bienzymatic strategy, based on a mediator-modified carbon film electrode and co-immobilisation of NADH oxidase and aldehyde dehydrogenase. Modification of the carbon film electrode with poly(neutral red) mediator resulted in a sensitive, low-cost and reliable NADH detector. Immobilisation of the enzymes was performed using encapsulation in a sol-gel matrix or cross-linking with glutaraldehyde. The bienzymatic biosensors were characterized by studying the influence of pH, applied potential and co-factors. The sol-gel and glutaraldehyde biosensors showed a linear response up to 60 μM and 100 μM, respectively, with detection limits of 2.6 μM and 3.3 μM and sensitivities were 1.7 μA mM⁻¹ and 5.6 μA mM⁻¹. The optimised biosensors showed good stability and good selectivity and have been tested for application for the determination of acetaldehyde in natural samples such as wine.     

Flow cell within an LED: a proposal for an optical absorption detector

Droplets formed at the tip of a tube under the same conditions possess extreme uniformity of form, volume and weight. These properties of liquid drop formation have been known for a long time and consequently many applications for the drop have been found in instrumentation and chemical analysis methods. In the present paper, we report on the analytical use of a dynamic LED-based flow-through optical absorption detector with optical path length controlled by continuous dropping of a solution. This arrangement consists of a flow cell built within a high-intensity red LED (λ _max = 630 nm). The feasibility of the detector is demonstrated by colorimetric determination of methylene blue, and ammonium by Berthelot’s reaction, in a flow-injection system. For ammonium, the reaction forms a blue dye (indophenol) with a maximum absorption at 630–650 nm. The detection limit, considered as 3 times the signal of the blank, is better than 125 μg l^-1. The small flow cell represents a good combination of optical path length, low volume and fast washout. This detector can be used advantageously in automated methods and can represent a solution to problems of optical detection involving gas bubbles and precipitation of particles in turbidimetric applications.     

Development of an optical chemical sensor based on 2-(5-bromo-2-pyridylazo)-5-(diethylamino)phenol in Nafion for determination of nickel ion

An optical chemical sensor based on immobilization of 2-(5-bromo-2-pyridylazo)-5-(diethylamino)phenol (Br-PADAP) in Nafion membrane is described. The membranes were cast onto glass substrates and were used for the determination of nickel in aqueous solutions by spectrophotometry. The sensor system is highly transparent, mechanically stable and showed no evidence of reagent leaching. The influence of several parameters such as pH, ligand concentration, and type and concentration of regenerating solution were optimized. The sensor system showed good sensitivity in the range 0.5-20 μg ml⁻¹ with a detection limit of 0.3 μg ml⁻¹ Ni(II). The sensor has been incorporated into a home-made flow-through cell for determination of nickel in flowing streams with improved sensitivity, precision and detection limit. The calibration curve in the flow system was linear in the range 0.1-16 μg ml⁻¹ with a detection limit of 0.07 μg ml⁻¹. The sensor is easily regenerated by dilute nitric acid solution. The proposed method was successfully applied to the determination of nickel content in vegetable oil and chocolate samples and the results were compared with those obtained using atomic absorption spectrometry.     

Fibrinolysis and thrombosis of fibrinogen-modified gold nanoparticles for detection of fibrinolytic-related proteins

Fibrinolysis (plasmin-mediated cleavage of fibrin structures) is a process in which fibrin clots can be removed from blood vessels, allowing the return of normal vascular function. Although several methods have been developed to measure plasmin activity and plasminogen (the plasmin precursor) concentrations, they are only moderately sensitive and quantitative and require large amounts of reagents, limiting their applicability. We developed two simple, label-free homogeneous assays using gold nanoparticles (Au NPs) for detection of fibrinolysis-related proteins and their activator (urokinase that converts plasminogen to plasmin) and inhibitor (α₂-plasmin inhibitor that inhibits plasmin and plasminogen bound to fibrin). We used a fibrinolysis-based sensor, based on plasmin-mediated cleavage of fibrinogen-modified Au NPs (Fib-Au NPs) leading to aggregation of Au NPs, to determine plasmin activity in a biological medium mimic solution. A combination of thrombin (Thr) and Fib-Au NPs allowed us to analyze plasmin activity and plasminogen concentrations in serum through Thr-induced agglutination of Fib-Au NPs. The limit of detection (LOD; S/N = 3) of this sensor for plasmin in serum was 0.4 nM (ca. 1.7 × 10⁻⁴ unit mL⁻¹). These label-free assays offer several advantages over conventional assays, including allowing rapid and simple readings with the naked eye or measurement by UV–vis absorption spectroscopy.     

Poly(ester imide)s possessing low coefficients of thermal expansion (CTE) and low water absorption (III). Use of bis(4-aminophenyl)terephthalate and effect of substituents

A variety of poly(ester imide)s (PEsIs) were prepared using bis(4-aminophenyl)terephthalate (BPTP) and substituted BPTP (BPTP series) for applications to novel base film materials in flexible printed circuit boards (FPC). BPTP series were all highly reactive with various tetracarboxylic dianhydrides and led to considerably high molecular weights of PEsI precursors. The thermally imidized BPTP-based PEsI films achieved lower extents of water absorption (W_A) than the corresponding 4-aminophenyl-4′-aminobenzoate (APAB)-based PEsI systems while keeping other target properties, in particular, the linear coefficient of thermal expansion (CTE) much lower than that of copper foil as a conductive layer in FPC. The lower W_A is attributed to the decreased imide contents in the structure by using BPTP. The considerably low CTE can be explained in terms of intimate stacking between the p-aromatic ester fragments with an extended conformation. The BPTP-based PEsI system also exhibited a considerably low dissipation factor (tan δ = 1.91 × 10⁻³) at a high-frequency electric field of 18.3 GHz, comparable to a liquid-crystalline polyester. An effect of substituents on the film properties was also investigated in this work. Incorporation of methyl substituents on BPTP was very effective for property improvement, whereas methoxy substituents on BPTP, as well as methyl substituents onto hydroquinone bis(trimellitate anhydride) (TAHQ), showed a trend to significantly increase the CTE. Copolymerization with an adequate amount of a typically flexible monomer, 4,4′-oxydianiline (4,4′-ODA), allowed the CTE matching with copper foil and the film toughness improvement at the same time. The PEsI copolymer prepared from TAHQ (10 mmol) with methyl-substituted BPTP (7 mmol) and 4,4′-ODA (3 mmol) achieved excellent combined properties, namely, a very high T_g at 410 °C, a slightly lower CTE (10.0 ppm/K) than that of copper foil, suppressed water absorption (0.35%), an extremely low linear coefficient of humidity expansion (CHE = 3.4 ppm/RH%), and good film toughness (the elongation at break, ε_b = 50.7%). Thus, BPTP- and methyl-substituted BPTP-based PEsI systems can be promising candidates as a next generation of FPC base film materials.     

Effects of substrate temperature on properties of ITO-ZnO composition spread films fabricated by combinatorial RF magnetron sputtering

We have fabricated ITO-ZnO composition spread films to investigate the effects of substrate temperature on their electrical and optical properties by using combinatorial RF magnetron sputtering. It turned out by X-ray measurement that the film with zinc contents above 16.0 at% [Zn/(In+Zn+Sn)] showed amorphous phase regardless of substrate temperature. The amorphous ITO-ZnO film had lower resistivity than polycrystalline films. When the films were deposited at 250 °C, the minimum resistivity of 3.0×10⁻⁴ Ω cm was obtained with the zinc contents of 16.0 at%. The indium content could be reduced as high as ~30 at% compared to that of ITO for the films having similar resistivity (~10⁻⁴ Ω cm). However, a drastic increase of resistivity was observed for the ITO-ZnO films deposited at 350 °C, having zinc contents below 15.2 at%.