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Biomedical subjects

Shouzhuo Yao

Publications and source records attributed to Shouzhuo Yao.

At least 19 recordsLinked to original sources

Electrochemical quartz crystal impedance study on the overoxidation of polypyrrole-carbon nanotubes composite film for amperometric detection of dopamine.

The electrochemical quartz crystal impedance (EQCI) method was used to study the overoxidation of polypyrrole (PPy)-multiwalled carbon nanotubes (MWCNT) nanocomposite film in neutral and alkaline solutions. The values of molar mass per electron transferred (M/n) obtained during the overoxidation of PPy in 0.10 mol L(-1) Na(2)SO(4) and 0.20 mol L(-1) NaOH aqueous solutions were estimated to be ca. 17 and 22 g mol(-1), respectively, suggesting the nucleophilic attack of solution OH(-) to the pyrrole units during the overoxidation, and the possible partial formation of carboxylic groups after the overoxidation in the NaOH solution. Also, the overoxidized PPy-MWCNT composite film prepared in the NaOH solution showed a notably larger affinity to dopamine (DA) dissolved in a neutral phosphate buffer than that prepared in the Na(2)SO(4) solution. The modification of the overoxidized nanocomposite film improved substantially the sensitivity for DA assay in a neutral phosphate buffer, as compared with the modification of overoxidized PPy or MWCNT alone. At a -6 kHz (201-nm thickness) nanocomposite film prepared in a polymerization bath containing 1.0 mg mL(-1) MWCNT and overoxidized in 0.20 mol L(-1) aqueous NaOH, the peak current response from differential pulse voltammetric assay of DA was linear with DA concentration from 4.0 x 10(-8) to 1.4 x 10(-6) mol L(-1), with a lower limit of detection of 1.7 nmol L(-1), good anti-interferent ability, as well as good stability and reproducibility.

Biosensing Techniques↗

A label-free electrochemical immunoassay for carcinoembryonic antigen (CEA) based on gold nanoparticles (AuNPs) and nonconductive polymer film.

A simple and sensitive label-free electrochemical immunoassay electrode for detection of carcinoembryonic antigen (CEA) has been developed. CEA antibody (CEAAb) was covalently attached on glutathione (GSH) monolayer-modified gold nanoparticle (AuNP) and the resulting CEAAb-AuNP bioconjugates were immobilized on Au electrode by electro-copolymerization with o-aminophenol (OAP). Electrochemical impedance spectroscopy and cyclic voltammetry studies demonstrate that the formation of CEA antibody-antigen complexes increases the electron transfer resistance of [Fe(CN)(6)](3-/4-) redox pair at the poly-OAP/CEAAb-AuNP/Au electrode. The use of CEA antibody-AuNP bioconjugates and poly-OAP film could enhance the sensitivity and anti-nonspecific binding of the resulting immunoassay electrode. The preliminary application of poly-OAP/CEAAb-AuNP/Au electrode for detection of CEA was also evaluated.

Biosensing Techniques↗

Polyoxometalate-modified carbon nanotubes: new catalyst support for methanol electro-oxidation.

A new catalyst support, polyoxometalate-modified carbon nanotubes, is presented in this paper through the chemisorption between polyoxometalate and carbon. Pt and Pt-Ru nanoparticles were electrochemically deposited on polyoxometalate-modified carbon nanotubes electrodes, and their electrocatalytic properties for methanol electro-oxidation are investigated in detail. Due to the unique electrical properties of carbon nanotubes and the excellent redox properties and the high protonic conductivity of polyoxometalate, for the similar deposition charge of Pt and Pt-Ru catalysts, 1.4 times larger exchange current density, 1.5 times higher specific activity, and better cycle stabilities can be obtained at polyoxometalate-modified carbon nanotube electrodes as compared to the electrodes without polyoxometalate modification. These results show that polyoxometalate-modified carbon nanotubes as a new catalyst support have good potential application in direct methanol fuel cells.

Journal Article↗

Determination of octopamine, synephrine and tyramine in Citrus herbs by ionic liquid improved 'green' chromatography.

Without adding any volatile organic solvents, aqueous solutions of room temperature ionic liquids (RTILs) were used as 'green' mobile phases to determine octopamine, synephrine and tyramine by liquid chromatography. The problems of the adrenergic amines separation, such as band tailing, low retention and low resolution were solved successfully by using RTIL. The effect of 1-ethyl-3-methylimidazolium tertafluoroborate ([EMIM][BF4]) was the best in the six investigated RTILs. The concentration of [EMIM][BF4], mobile phase pH and column temperature, which influenced the chromatographic behaviors of the analytes, were investigated in detail. The change of retention factors caused by pH shift was obviously suppressed by [EMIM][BF4]. The sensitivity, accuracy and repeatability of this method were found to be satisfactory. The contents of adrenergic amines in several Citrus herbs and extracts, such as Fructus aurantii immaturus, were simultaneously determined by this 'green' chromatographic method.

Adrenergic alpha-Agonists↗

Monitoring and estimation of the kinetics parameters in the binding process of tannic acid to bovine serum albumin with electrochemical quartz crystal impedance system.

Combined measurements of piezoelectric quartz crystal impedance (PQCI) and electrochemical impedance (EI) were utilized in situ to monitor the adsorption of bovine serum albumin (BSA) onto the newly prepared Au colloid-modified electrode and study the binding process of tannic acid (TA) to BSA on the BSA-modified electrode surface. The time courses of the resonant frequency and the equivalent parameters of the sensor were simultaneously obtained during BSA adsorption and TA-BSA binding. Compared with the bare gold electrode, the Au colloid-modified gold electrode showed better biocompatibility, and the absorption capacity for BSA was increased by approximately 2.4 times. The observed frequency decrease was ascribed to the mass increase of the sensor surface resulting from the TA-BSA binding, which is believed to result mainly from the hydrogen bonding from FT-IR characterization. The maximal molar binding ratio of TA binding to immobilized BSA obtained from the frequency shift of the adsorbed BSA and TA was estimated to be 10.3:1. On the basis of the frequency decrease with time, the kinetics of the binding was quantitatively studied. By way of fitting the experimental data, the kinetics parameters, that is, binding and dissociation constant (k1, k(-1)), and the binding equilibrium constant (ka) were determined, giving values of 9.51 x 10(4) M(-1) s(-1), 3.15 s(-1), and 3.1 x 10(4) M(-1), respectively.

Electric Impedance↗

Electrochemical piezoelectric quartz crystal impedance study on the interaction between concanavalin A and glycogen at Au electrodes.

The carbohydrate research has emerged as a "new frontier" in chemical/biological field. The binding of lectin with carbohydrate is one of the important courses of life activities. The report studies concanavalin A (Con A)-glycogen interaction on gold electrode surfaces by electrochemical piezoelectric quartz crystal impedance (EPQCI) method. The piezoelectric quartz crystal (PQC) parameters, resonant frequency shift (Deltaf(0)) and the motional resistance change (DeltaR(1)), and the electrochemical impedance (EI) parameters, electrolyte resistance change (DeltaR(s)) and the double layer capacitance change (DeltaC(s)), were measured and discussed simultaneously. Two methods were adopted for measuring the Con A-glycogen association. Based on EPQCI measurement during Con A reaction with glycogen adsorbed on Au electrode, association constant K(a) and the amount of the binding sites s calculated are 1.48 x 10(6) M(-1) and 4.09, respectively. Based on single PQC measurement of glycogen reaction with Con A assembled on Au electrode, K(a) was estimated to be 1.26 x 10(6) M(-1).

Biosensing Techniques↗

Comparative study on the interaction of DNA with three different kinds of surfactants and the formation of multilayer films.

The interactions between double-stranded DNA (dsDNA) and three different kinds of surfactants, i.e., cationic, anionic, and nonionic surfactants, were investigated by cyclic voltammetry, electrochemical impedance spectroscopy and UV-vis spectroscopy. Multilayer films composed of DNA and surfactants were prepared at gold electrode by electrostatic or hydrophobic interactions. It was found that the cationic surfactant, CTAB, can bind to DNA by electrostatic interaction, and the electron transfer resistance of CTAB-DNA complex film increases first and then decreases with CTAB concentration. The anionic surfactant, LAS, can bind to DNA but by hydrophobic interaction, and the electron transfer resistance of the complex film keeps decreasing with LAS concentration. Nonionic surfactants can also directly bind to DNA by hydrophobic interaction. All the three different kinds of surfactants can form multilayer films with DNA on the electrode surface. The chemical structure of DNA keeps unchanged during interacting with these surfactants. The binding modes of DNA with these three different kinds of surfactants were also deduced.

Binding Sites↗

Determination of angiotensin converting enzyme inhibitory activity by high-performance liquid chromatography/electrospray-mass spectrometry.

A sensitive and rapid method for determination of angiotensin converting enzyme (ACE) inhibitory activity was developed based on a combination of enzymatic reaction followed by high performance liquid chromatography/electrospray-mass spectrometry (HPLC-ESI-MS) determination of its product. The most commonly used substrate hippuryl-histidyl-leucine (HHL) or hippuryl-glycyl-glycine (HGG) hydrolysis catalyzed by purified rabbit lung ACE or human plasma ACE was investigated in the presence of benazeprilat. The incubation time was 8 min for purified lung ACE, and 16 min for human plasma ACE. The produced hippuric acid (HA) was separated from substrate HHL or HGG by HPLC on a C(18) column with isocratic elution within 6.5 min, and quantified by electrospray ionization mass spectrometry (ESI-MS) with p-phthalic acid as an internal standard (IS). The limit of detection of HA was 6.0 ng/ml. HHL or HGG hydrolysis catalyzed by purified lung ACE displayed excellent accuracy and reproducibility. The small total reaction volume, the low concentration of substrate, and the simple treating procedures present the advantages of the new method. Furthermore, the total time of the whole procedure for one sample with the novel method is less than 1/2 of that of the conventional HPLC or spectrophotometry method, while the accuracy and the precision of the new method are almost the same as the conventional HPLC method with UV detection.

Angiotensin-Converting Enzyme Inhibitors↗

High-performance liquid chromatography-electronspray ionization mass spectrometry for determination of tiopronin in human plasma.

A simple and sensitive HPLC/ESI-MS method for the determination of tiopronin in human plasma was described. Vitamin C and 2-mercaptoethanol (2-Me) were used as the reducer and the stabilitizer to release and stabilify tiopronin from a dimmer and mix forms with endogenous thiols in the treatment of plasma samples. The analytes were separated on a Johnson Spherigel analytical column packed with 5 microm C8 silica, using the formic acid aqueous solution (pH 4.5) including tris(hydroxymethyl) aminomethane (Tris) and 2-Me (0.5 and 1 mM, respectively) as a mobile phase. Cyclamate was used as the internal standard (I.S.) for the quantification of tiopronin. The correlation coefficient of the calibration curve were better than 0.998 in the range of 0.107-5.35 microg/ml in human plasma. The limit of quantification (LOQ) was 0.107 microg/ml (S/N 10:1, RSD 7.1%). The inter-day and intra-day accuracy was below 7.1 and 6.8%, respectively. As a preliminary application, this method has been successfully applied to the determination of tiopronin in the human plasma.

Calibration↗

Nonaqueous capillary electrophoresis coupled with laser-induced native fluorescence detection for the analysis of berberine, palmatine, and jatrorrhizine in Chinese herbal medicines.

LIF detection is one of the most sensitive detection methods for CE. However, its application is limited because the analyte is usually required to be derivatized with a fluorescent label. As a result, LIF is seldom used to analyze active ingredients in plants. In this work, we introduce a rapid, simple, and sensitive method of nonaqueous CE (NACE) coupled with laser-induced native fluorescence detection for the simultaneous analysis of berberine, palmatine, and jatrorrhizine. This method skillfully utilizes the native fluorescence of these alkaloids and requires no troublesome fluorescent derivatization. As these alkaloids can fluoresce to some degree, they were simply detected by a commercially available 488 nm Ar+ laser. The native fluorescence of the analytes was greatly enhanced by nonaqueous media. Compared with the reported UV detection method, much lower LOD was achieved (6.0 ng/mL for berberine, 7.5 ng/mL for palmatine, and 380 ng/mL for jatrorrhizine). This method was successfully applied to analyze berberine, palmatine, and jatrorrhizine in two Chinese herbal medicines, Rhizoma coptidis and Caulis mahoniae.

Acetates↗

Determination of difenidol hydrochloride by capillary electrophoresis with electrochemiluminescence detection.

A novel and sensitive method for the determination of difenidol hydrochloride has been established using capillary electrophoresis coupled with end-column electrogenerated chemiluminescence (ECL) detection, based on the ECL reaction of tris(2,2'-bypyridine)ruthenium(II) (Ru(bpy)(3)(2+)) with the tertiary amino groups of the difenidol analyte. Parameters that affect separation and detection were optimized. Calibration curve was linear over the range from 1 x 10(-6)M to 6 x 10(-5)M with a detection limit of 1 x 10(-7)M (S/N=3). Separation of difenidol hydrochloride from clomifene citrate and lidocaine was achieved using the proposed method. This method was successfully utilized to the assay of the active ingredients of the "difenidol hydrochloride" tablets and to the investigation on the interaction of difenidol hydrochloride with hemoglobin. The number of binding sites and the binding constant were estimated as (11.2 and 2.5) x 10(3)M(-1), respectively.

2,2'-Dipyridyl↗

Monitoring of cell growth and assessment of cytotoxicity using electrochemical impedance spectroscopy.

Electrochemical impedance spectroscopy (EIS) was used to monitor the growth of mammalian cancer cells and evaluate the cytotoxicity of chemicals using Fe(CN)6(3-/4-) as a redox probe. Cancer cells, the human hepatocarcinoma cell line (BEL7404), were grown on optically transparent indium tin oxide (ITO) semiconductor slides, which were used as the working electrodes in electrochemical experiments. Attachment and proliferation of cancer cells on ITO surfaces resulted in increase of electron-transfer resistance (R(et)) between the redox probe of Fe(CN)6(3-/4-) in electrolyte solution and ITO electrode surface. For cytotoxicity assessment, cells grown on ITO substrates were further cultured in the presence of different cytotoxicants and electrochemical impedance measurements were carried out at different time intervals. Gemcitabine, a promising antineoplastic drug showing activity against a wide spectrum of human solid tumors, was selected as a model for long-term cytotoxicity effect study, whereas mercury chloride represented a model for acute toxicants. The inhibitions of gemcitabine and mercury chloride on the viability and proliferation of BEL7404 cells were observed from the electrochemical impedance experiments, and the different action modes were discriminated. Additionally, microscope images were also used to observe the effects of these two chemicals on the morphology of the cells. General consistency has been found between the electrochemical impedance response and the morphological observation. Such an impedance method provides a simple and inexpensive way for in vitro assessment of chemical cytotoxicity.

Antineoplastic Agents↗

Simultaneous determination of water-soluble and fat-soluble synthetic colorants in foodstuff by high-performance liquid chromatography-diode array detection-electrospray mass spectrometry.

An accurate method was developed for the simultaneous determination of water-Tartrazine, Amaranth, Ponceau 4R, Sunset Yellow FCF, and fat-Sudan (I-IV), synthetic soluble colorants in foodstuff. This method uses dimethylsulfoxide (DMSO) as the extraction solvent in the sample preparation process and high performance liquid chromatography (HPLC)-diode array detector (DAD)-electrospray mass spectrometry (ESI-MS), applying selected ion recording in positive/negative alternate mode to acquire mass spectral data, as the analytical technique. Linearity of around three orders in the magnitude of concentration was generally obtained. Detection and quantification limits of the investigated dyes, which were evaluated at signal to noise ratio of 3 for detection limit and 10 for quantification limit, were in the ranges of 0.01-4 and 0.03-11.2 ng, respectively. The recoveries of the eight synthetic colorants in four matrices ranged from 93.2 to 108.3%. Relative standard deviations of less than 8.2% were also achieved. This method has been applied successfully in the determination of water-soluble colorants in the soft drink and the delicious ginger, and fat-soluble dyes in chilli powders and chilli spices.

Beverages↗

An electrochemical quartz crystal impedance study on anti-human immunoglobulin G immobilization in the polymer grown during dopamine oxidation at an Au electrode.

The polymeric film grown during dopamine oxidation at an Au electrode was studied as a novel matrix for immobilizing anti-human immunoglobulin G (IgG) via the electrochemical quartz crystal impedance analysis (EQCIA) method. The growth of the polymeric films at Au electrodes during dopamine oxidation in neutral phosphate buffer (pH 7.4) and the immobilization of anti-human IgG into the polymeric films during their growth have been traced at real time. Lysozyme control experiments suggested that anti-human IgG was electrostatically incorporated into the polymeric film. Also, the porosity of the polymeric films has been discussed by measuring the "wet" and "dry" frequency shifts. Compared with a polypyrrole film immobilized with anti-human IgG, the proposed matrix possessed a larger amount of specific binding sites for human IgG by subsequent immunoreaction tests. The association constant of the anti-human IgG immunoreaction was obtained with satisfactory results.

Crystallization↗

Determination of tetramethylenedisulfotetramine in human urine with gas chromatograph-flame thermionic detection coupling with direct immersed solid-phase micro-extraction.

An analytical method for determination of tetramethylenedisulfotetramine (tetramine) in human urine by gas chromatography-flame thermionic detection (GC-FTD) coupling with a direct immersed solid-phase micro-extraction (DI-SPME) was developed. The enrichment effects of three fiber coatings of SPME for tetramine were compared. Results showed that the enrichment effect of polar 85 microm polyacrylate (PA) coating was better than that of apolar 100 and 7 microm polydimethylsiloxane (PDMS) coatings. Other experimental parameters, such as ionic strength, volume, temperature of sample solution and time for extraction, time and temperature for desorption, were also optimized. The correlation coefficient of the calibration curve was 0.9998 in the range of 0.082-41.0 ng/mL for tetramine. The limit of quantitation of tetramine in urine was 0.082 ng/mL. In this method, the sample pretreatment is simple and convenient. As a monitoring means, it has been successfully applied to detection of tetramine toxicosis in criminal cases, as well as clinical therapy of poisoned sufferer.

Bridged-Ring Compounds↗

Separation and purification of chlorogenic acid by molecularly imprinted polymer monolithic stationary phase.

Separation and purification of chlorogenic acid by removal of the impurities compounds co-existed in the product using molecular imprinting technique was firstly reported. In this work, an in situ synthesis method was utilized for the preparation of molecularly imprinted polymer monolithic stationary phase using the impurity molecule (caffeic acid) as template, the mixture of tetrahydrofuran and isooctane as solvent, and methacrylic acid and ethyl glycol dimethacrylate as functional monomer and cross-linker, respectively. The retention behavior of the monolithic polymer to chlorogenic acid molecule, the template and several main impurities compounds in the product was studied and the adsorption capacity of compounds on the stationary phase determined by frontal chromatographic technique. A relatively weak retention of the target product molecule (chlorogenic acid) on the polymer and a strong adsorption capability of the monolith to the template and several main impurities were observed. This might mainly result from the 'shape' difference of chlorogenic acid molecule with the impurities compounds molecules. This approach was shown to be successful for the separation and purification of chlorogenic acid from the extract of Eucommia ulmodies leaves.

Adsorption↗

DNA biosensor based on chitosan film doped with carbon nanotubes.

A biosensor based on chitosan doped with carbon nanotube (CNT) was fabricated to detect salmon sperm DNA. Methylene blue (MB) was employed as a DNA indicator. It was found that CNTs can enhance the electroactive surface area threefold (0.28 +/- 0.03 and 0.093 +/- 0.06 cm(2) for chitosan-CNT- and chitosan-modified electrodes, respectively) and can accelerate the rate of electron transfer between the redox-active MB and the electrode. A low detection limit of 0.252 nM fish sperm DNA was achieved, and no interference was found in the presence of 5 microg/ml human serum albumin. The differential pulse voltammetry signal of MB was linear over the fish sperm DNA concentration range of 0.5-20 nM.

Animals↗

Liquid-phase microextraction combined with high-performance liquid chromatography for the determination of local anaesthetics in human urine.

A simple liquid-phase microextraction (LPME) device combined with high-performance liquid chromatography (HPLC) is presented for the simultaneous analysis of local anaesthetics, lidocaine, bupivacaine, and tetracaine, from human urine sample. An organic solvent showed good compatibility with the mobile phase of the HPLC, o-dibutyl phthalate, was selected. Local anaesthetics are extracted from 6 ml of the feed aqueous solution and human urine sample into a water-immiscible organic solvent suspended at the needle tip of the microsyringe, then the organic solvent was directly introduced to a reversed-phase HPLC system. The kind of the organic extraction solvent, the stirring rate, the pH value of the aqueous feed solution, and the extraction time have been discussed. Under the optimized extraction conditions, high enrichment factors (more than 86.0-fold) and significant sample clean-up for all of studied local anaesthetics were achieved within 30 min. The detection limits (lower than 0.05 microg/ml) were comparable with previously reported gas chromatography methods. This method was applied to specimen of patient who was treated with extradural anaesthesia of lidocaine, bupivacaine, and tetracaine, and revealed that simultaneous determination of above three local anaesthetics in human urine was possible.

Anesthetics, Local↗