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1.
This work reported for the first time the use of flow injection electrochemiluminescence (FI-ECL) sensor for the determination
of durabolin in an aqueous system based on CdTe quantum dot (QD) films. Aqueous CdTe colloidal solutions were prepared using
thioglycolic acid as a capping agent. Zetasizer Nano ZS (Malvern, UK) was employed to characterize the size of CdTe QDs. The
UV–vis and photoluminescence spectra of samples were systematically characterized. Indium tin oxide (ITO) slide glass was
modified with CdTe QDs by layer-by-layer self-assembly. CdTe QD films were packed into a homemade cell and used as a recognizer
of the FI-ECL sensor to determine durabolin. The intensive anodic ECL emission was obtained at a starting potential of +1.3 V
(vs. Ag/AgCl) in a carbonate bicarbonate buffer solution with a pH of 9.93 at an ITO electrode. The ECL intensity was correlated
linearly with the concentration of durabolin over the range of 1.0 × 10−8–1.0 × 10−5 g mL−1, and the detection limit was 2.5 × 10−9 g mL−1. The relative standard deviation for the determination of 1.0 × 10−6 g mL−1 durabolin was 1.04% (n = 11). This simple and sensitive sensor revealed good reproducibility for ECL analysis. As a result, the new FI-ECL sensor
had been successfully applied to the determination of durabolin in food samples. This strategy could be easily realized and
opened new avenues for the applications of QDs in ECL biosensing. 相似文献
2.
Gema Paniagua González Pilar Fernández Hernando Jesús Senén Durand Alegría 《Analytical and bioanalytical chemistry》2009,394(4):963-970
This work reports a comparative study of two automated flow-through fluorosensors for the determination of digoxin in serum
samples: an immunosensor with an anti-digoxin polyclonal antibody as the reactive phase permanently immobilised on controlled-pore
glass and a sensor with a selective reaction system based on a methacrylic molecularly imprinted polymer (MIP) synthesised
by bulk polymerisation. The variables affecting the sensitivity and dynamic range of the sensors (e.g. the carrier and elution
solutions, flow rates, pH and reagent concentrations) were optimized, and the binding characteristics of their reactive phases
were compared in a competitive fluorescent assay. Digoxin was reproducibly determined by both sensors at the milligram per
litre level (detection limit = 1.20 × 10−3 mg L−1 and RSD = 4–7% for the immunosensor; detection limit = 1.7 × 10−5 mg L−1 and RSD = 1–2% for the MIP sensor). No cross-reactivity with digoxin-related compounds was seen for either sensor at a digoxin/interferent
ratio of 1:100. The lifetime of the immunosensor was about 50 immunoassays; its shelf life, when unused, is about 3 months.
The lifetime of the MIP sensor was over 18 months. Both sensors were used to determine the digoxin concentration of human
serum samples with satisfactory results. 相似文献
3.
CdTe quantum dots (QDs) were modified with thioglycolic acid (TGA) and synthesized in aqueous medium. The optimum fluorescence
intensity was found to be at pH 6.24 with a CdTe QDs concentration of 4.96 × 10−7 mol L−1. The quenched fluorescence intensity of CdTe QDs is linearly proportional to V(V) concentration from 10 to 200 ng mL−1 with correlation coefficient R = 0.9985. The limit of detection for V(V) was 2.07 ng mL−1. The proposed method was successfully applied to the analysis of trace amounts of V(V) in water samples with recovery of
96.5–101.8%, and the results were in good agreement with those of electrothermal atomic absorption spectrometry. 相似文献
4.
A fast and sensitive liquid chromatography–mass spectrometry method was developed for the determination of ursolic acid (UA)
in rat plasma and tissues. Glycyrrhetinic acid was used as the internal standard (IS). Chromatographic separation was performed
on a 3.5 μm Zorbax SB-C18 column (30 mm × 2.1 mm) with a mobile phase consisting of methanol and aqueous 10 mM ammonium acetate
using gradient elution. Quantification was performed by selected ion monitoring with (m/z)− 455 for UA and (m/z)− 469 for the IS. The method was validated in the concentration range of 2.5 − 1470 ng mL−1 for plasma samples and 20 − 11760 ng g−1 for tissue homogenates. The intra- and inter-day assay of precision in plasma and tissues ranged from 1.6% to 7.1% and 3.7%
to 9.0%, respectively, and the intra- and inter-day assay accuracy was 84.2 − 106.9% and 82.1 − 108.1%, respectively. Recoveries
in plasma and tissues ranged from 83.2% to 106.2%. The limits of detections were 0.5 ng mL−1 or 4.0 ng g−1. The recoveries for all samples were >90%, except for liver, which indicated that ursolic acid may metabolize in liver. The
main pharmacokinetic parameters obtained were T
max = 0.42 ± 0.11 h, C
max = 1.10 ± 0.31 μg mL−1, AUC = 1.45 ± 0.21 μg h mL−1 and K
a = 5.64 ± 1.89 h−1. The concentrations of UA in rat lung, spleen, liver, heart, and cerebellum were studied for the first time. This method
is validated and could be applicable to the investigation of the pharmacokinetics and tissue distribution of UA in rats. 相似文献
5.
Determination of salbutamol using R-phycoerythrin immobilized on eggshell membrane surface as a fluorescence probe 总被引:1,自引:0,他引:1
Jieli Tang Zhenshuang Liu Jing Kang Yihua Zhang 《Analytical and bioanalytical chemistry》2010,397(7):3015-3022
A fluorescence sensor was fabricated using R-phycoerythrin (R-PE) immobilized on eggshell membrane as the fluorescence probe,
and salbutamol was determined based on the decrease in fluorescence intensity of R-phycoerythrin. The scanning electron and
fluorescence micrographs showed the microstructure of the eggshell membrane and indicated that the R-PE was successfully immobilized
on the eggshell membrane surface. The effects of some experimental parameters on the response of the biosensor were investigated
in detail. The fluorescence sensor has a linear response to salbutamol concentrations ranging from 5.00 to 100 ng mL−1. The detection limit for the salbutamol is 3.50 ng mL−1 (S/N = 3). The reproducibility of fabricating the biosensors using six different membranes was good with a relative standard
deviation (RSD) of 3.28%. The fluorescence sensor showed extremely good stability with a shelf life of at least 50 days and
reversible response to salbutamol. Some common potential interferents showed little effect on the response of the salbutamol
fluorescence sensor. The proposed method was successfully applied to the determination of the salbutamol in urine samples. 相似文献
6.
Bagheri H Aghakhani A Akbari M Ayazi Z 《Analytical and bioanalytical chemistry》2011,400(10):3607-3613
A micro-solid phase extraction technique was developed using a novel polypyrrole-polyamide nanofiber sheet, fabricated by
electrospinning method. The applicability of the new nanofiber sheet was examined as an extracting medium to isolate malathion
as a model pesticide from aqueous samples. Solvent desorption was subsequently performed in a microvial, and an aliquot of
extractant was injected into gas chromatography–mass spectrometry. Various parameters affecting the electrospinning process
including monomer concentration, polyamide content, applied voltage, and electrospinning time were examined. After fabricating
the most suitable preparation conditions, influential parameters on the extraction and desorption processes were optimized.
The developed method proved to be rather convenient and offers sufficient sensitivity and good reproducibility. The limit
of detection (S/N = 3) and limit of quantification (S/N = 10) of the method under optimized conditions were 50 and 100 ng L−1, respectively. The relative standard deviation at concentration level of 1 ng mL−1 was 2% (n = 3). The calibration curve of analyte showed linearity in the range of 0.1–1 ng mL−1 (R
2 = 0.9975). The developed method was successfully applied to tap and Zayanderood river water samples, while the relative recovery
percentages of 98% and 96% were obtained, respectively. The whole procedure showed to be conveniently applicable and quite
easy to be manipulated. 相似文献
7.
Abdel-Aziz Youssef El-Sayed Ebtesam Ahmad Saad Basheer Mohamed Mohamed Ibrahime Mohamed Tarek Mohamed Zaki 《Mikrochimica acta》2000,135(1-2):19-27
Simple, rapid, sensitive and selective methods for the determination of Cr(III) and W(VI) with flavonol derivatives in the
presence of surface-active agents are proposed. In the pH ranges 3.4–4.2 and 1.9–2.5, the molar absorptivities of Cr(III)-morin-emulsifier
S (EFA) and W(VI)-morin-polyvinylpyrrolidone (PVP) systems are 1.13×105 and 2.13×104 L mol−1 cm−1 at 435 and 415 nm, respectively. The Cr(III)-quercetin-PVP and W(VI)-quercetin-cetylpyridinium bromide (CPB) systems are
formed in the pH ranges 4–4.6 and 2.2–2.8 with molar absorptivities 1.02×105 and 9.02×104 L. mol−1 cm−1 at 441 and 419 nm, respectively. The linear dynamic ranges for the determination of Cr(III) and W(VI) with morin in the presence
of EFA and PVP are 0.03–0.46 and 0.71–8.1 μg mL−1, respectively. The corresponding ranges with quercetin are 0.04–0.54 and 0.14–2.1 μg mL−1 of Cr(III) and W(VI), respectively. The r.s.d (n = 10) for the determination of 0.25 and 3.7 μg mL−1 of Cr(III) and W(VI) with morin and their detection limits are 0.88 and 0.99% and 0.016 and 0.63 μg mL−1, respectively. Using quercetin, the r.s.d (n = 10) for 0.22 and 1.2 μg mL−1 of Cr(III) and W(VI) and their detection limits are 0.92 and 0.91% and 0.015 and 0.08 μg mL−1, respectively. The critical evaluation of the proposed methods is performed by statistical analysis of the experimental data.
The proposed methods are applied to determine Cr in steel, non-ferrous alloys, wastewater and mud filtrate and to the determination
of W in steel.
Received March 8, 1999. Revision January 21, 2000. 相似文献
8.
This work presents a comparison of convective interaction media (CIM) and controlled pore glass (CPG) as solid supports for
immunoglobulin antibodies used in bioanalytical detection of allergens in foodstuffs. A flow-injection manifold with highly
sensitive thermal lens spectrometric detection was used for this purpose. Using beta-lactoglobulin, a milk allergen, as a
model analyte, CIM disc supports had a higher linear range (0.2–3.5 μg L−1), better reproducibility (intra-day RSD = 1%, inter-day RSD = 10%), lower consumption of reagents, and better immunocolumn
stability (1 month, over 240 injections of substrate), while providing comparable LODs (0.1 μg L−1). Application of CIM discs as solid supports in immunocolumns for allergen detection enables fast and sensitive screening
of allergens in foodstuffs with sample throughput of up to eight samples per hour. 相似文献
9.
Xiaoying Liu Yonghong Li Xinsheng Liu Xiandong Zeng Bo Kong Shenglian Luo Wanzhi Wei 《Journal of Solid State Electrochemistry》2012,16(3):883-889
A simple sensor based on bare carbon ionic liquid electrode was fabricated for simultaneous determination of dihydroxybenzene
isomers in 0.1 mol L−1 phosphate buffer solution (pH 6.0). The oxidation peak potential of hydroquinone was about 0.136 V, catechol was about 0.240 V,
and resorcinol 0.632 V by differential pulse voltammetric measurements, which indicated that the dihydroxybenzene isomers
could be separated absolutely. The sensor showed wide linear behaviors in the range of 5.0 × 10−7–2.0 × 10−4 mol L−1 for hydroquinone and catechol, 3.5 × 10−6–1.535 × 10−4 mol L−1 for resorcinol, respectively. And the detection limits of the three dihydroxybenzene isomers were 5.0 × 10−8, 2.0 × 10−7, 5.0 × 10−7 mol L−1, respectively (S/N = 3). The proposed method could be applied to the determination of dihydroxybenzene isomers in artificial wastewater and
the recovery was from 93.9% to 104.6%. 相似文献
10.
Marcela Z. Arend Simone G. Cardoso Felipe K. Hurtado Aline Ravanello Fibele A. Lanzanova Clarice M. B. Rolim 《Chromatographia》2009,69(Z2):195-199
A simple stability-indicating reversed-phase liquid chromatographic method with diode-array detection was developed and validated
for the quantitative determination of ebastine in tablets and syrup. The LC method was carried out on a C18 column with acetonitrile:phosphoric acid 0.1% pH 3.0 (55:45, v/v) as mobile phase, at a flow rate of 1.2 mL min−1. Ultraviolet detection of ebastine was at 254 nm. A linear response (r = 0.9999) was observed in the range of 10–80 μg mL−1. The RSD values for intra- and inter-day precision studies showed good results (RSD < 2%) and accuracy was greater than 98%.
Validation parameters such as specificity and robustness were also determined. The method was found to be stability-indicating
and can be applied to quantitative determination of ebastine in tablets and syrup. 相似文献
11.
Borges KB de Oliveira AR Barth T Jabor VA Pupo MT Bonato PS 《Analytical and bioanalytical chemistry》2011,399(2):915-925
The purpose of this study was the development and validation of an LC–MS–MS method for simultaneous analysis of ibuprofen
(IBP), 2-hydroxyibuprofen (2-OH-IBP) enantiomers, and carboxyibuprofen (COOH-IBP) stereoisomers in fungi culture medium, to
investigate the ability of some endophytic fungi to biotransform the chiral drug IBP into its metabolites. Resolution of IBP
and the stereoisomers of its main metabolites was achieved by use of a Chiralpak AS-H column (150 × 4.6 mm, 5 μm particle
size), column temperature 8 °C, and the mobile phase hexane–isopropanol–trifluoroacetic acid (95: 5: 0.1, v/v) at a flow rate of 1.2 mL min−1. Post-column infusion with 10 mmol L−1 ammonium acetate in methanol at a flow rate of 0.3 mL min−1 was performed to enhance MS detection (positive electrospray ionization). Liquid–liquid extraction was used for sample preparation
with hexane–ethyl acetate (1:1, v/v) as extraction solvent. Linearity was obtained in the range 0.1–20 μg mL−1 for IBP, 0.05–7.5 μg mL−1 for each 2-OH-IBP enantiomer, and 0.025–5.0 μg mL−1 for each COOH-IBP stereoisomer (r ≥ 0.99). The coefficients of variation and relative errors obtained in precision and accuracy studies (within-day and between-day)
were below 15%. The stability studies showed that the samples were stable (p > 0.05) during freeze and thaw cycles, short-term exposure to room temperature, storage at −20 °C, and biotransformation
conditions. Among the six fungi studied, only the strains Nigrospora sphaerica (SS67) and Chaetomium globosum (VR10) biotransformed IBP enantioselectively, with greater formation of the metabolite (+)-(S)-2-OH-IBP. Formation of the COOH-IBP stereoisomers, which involves hydroxylation at C3 and further oxidation to form the
carboxyl group, was not observed. 相似文献
12.
Cooper EM Covaci A van Nuijs AL Webster TF Stapleton HM 《Analytical and bioanalytical chemistry》2011,401(7):2123-2132
Organophosphate triesters tris(1,3-dichloro-2-propyl) phosphate (TDCPP) and triphenyl phosphate are widely used flame retardants
(FRs) present in many products common to human environments, yet understanding of human exposure and health effects of these
compounds is limited. Monitoring urinary metabolites as biomarkers of exposure can be a valuable aid for improving this understanding;
however, no previously published method exists for the analysis of the primary TDCPP metabolite, bis(1,3-dichloro-2-propyl)
phosphate (BDCPP), in human urine. Here, we present a method to extract the metabolites BDCPP and diphenyl phosphate (DPP)
in human urine using mixed-mode anion exchange solid phase extraction and mass-labeled internal standards with analysis by
atmospheric pressure chemical ionization liquid chromatography tandem mass spectrometry. The method detection limit was 8 pg mL−1 urine for BDCPP and 204 pg mL−1 for DPP. Recoveries of analytes spiked into urine ranged from 82 ± 10% to 91 ± 4% for BDCPP and from 72 ± 12% to 76 ± 8%
for DPP. Analysis of a small number of urine samples (n = 9) randomly collected from non-occupationally exposed adults revealed the presence of both BDCPP and DPP in all samples.
Non-normalized urinary concentrations ranged from 46–1,662 pg BDCPP mL−1 to 287–7,443 pg DPP mL−1, with geometric means of 147 pg BDCPP mL−1 and 1,074 pg DPP mL−1. Levels of DPP were higher than those of BDCPP in 89% of samples. The presented method is simple and sufficiently sensitive
to detect these FR metabolites in humans and may be applied to future studies to increase our understanding of exposure to
and potential health effects from FRs. 相似文献
13.
Ying Xu Fei Wang Le Wang Fangyuan Zhao Baocheng Yang Baoxian Ye 《Journal of Solid State Electrochemistry》2012,16(4):1473-1480
A novel voltammetric sensor, based on single-walled carbon nanotubes (SWNT) dispersed in Nafion and modified glassy carbon
electrode (GCE), was fabricated and used to determine the trace amounts of dihydromyricetin (DMY). The electrochemical behavior
of DMY at this sensor was investigated in 0.1 mol L−1 sulfuric acid solutions + 0.1 mol L−1 NaCl by cyclic voltammetry and squarewave voltammetry. Compared with bare GCE, the electrode presented an excellent response
of DMY through an adsorption-controlled quasi-reversible process. Under the optimum conditions, the response peak currents
were linear relationship with the DMY concentrations in the range of 1.0 × 10−7–1.0 × 10−5 mol L−1 with a detection limit of 9 × 10−8 mol L−1. Based on this voltammetric sensor, a simple and sensitive electroanalytical method for DMY was proposed and applied to quantitative
determination of DMY in Ampelopsis grossedentata samples. In addition, the oxidation mechanism was proposed and discussed, which could be a reference for the pharmacological
action of DMY in clinical study. 相似文献
14.
It was found that tripropylamine (TPA) could be used as a coreactant to initiate the electrochemiluminescence (ECL) of acridinium
NHS ester (AE NHS) labels attached to DNA. The radicals generated in the electro-oxidation process of TPA reacted with AE
NHS to form the excited N-methylacridone, giving rise to light emission. The AE/TPA ECL system was for the first time used as the detection system
for developing an ECL-based DNA sensor. In the protocol, streptavidin-modified gold nanoparticles were firstly immobilized
onto a thiol-treated gold electrode. The streptavidin could specifically interact with the biontinylated capture DNA. Afterwards,
the target DNA and the AE-labeled report DNA were conjugated onto the electrode step by step due to the hybridization reactions,
and a sandwich-type sensor was fabricated. The ECL signals of the sensor were obtained under pulse potential condition in
alkaline solution containing 50.0 mmol L−1 TPA. Under optimized experimental conditions, the linear range of the DNA sensor for the determination of the target DNA
was from 5.0 × 10−15 to 5.0 × 10−12 mol L−1. The detection limit (S/N = 3) was 3.0 × 10−15 mol L−1. Moreover, the sensor could specifically recognize the target DNA against one base-pair mismatched sequences, two base-pair
mismatched sequences, and the noncomplementary sequences. It is of great application potential in clinic analysis. 相似文献
15.
We have developed a circular-dichroism thermal lens microscope for UV wavelengths (UV-CD-TLM), for the first time, to realize
sensitive chiral analysis on a microchip. Quasi-continuous-wave phase modulation of a pulsed UV laser was used to generate
left-circularly polarized light and right-circularly polarized light and to detect the generated TL signal amplitude and phase
with a lock-in amplifier. The amplitude and phase were used to determine the concentration and chirality, respectively, of
a sample. The basic principle of UV-CD-TLM for chiral analysis on a microchip was verified by measuring aqueous solutions
of optically active camphorsulfonic acids (CSA). Lower limits of detection (LOD) were calculated at S/N = 2 and were 8.7 × 10−4 mol L−1 (ΔA = 5.2 × 10−6 Abs.) for (+)-CSA and 8.4 × 10−4 mol L−1 (ΔA = 5.0 × 10−6 Abs.) for (−)-CSA. In terms of number of molecules, LODs for UV-CD-TLM were calculated to be 8.7 fmol and 8.4 fmol, respectively.
This is at least three orders of magnitude lower than previously obtained. The applicability of UV-CD-TLM for chiral analysis
on a microchip was verified.
Figure Sensitive chiral analysis by thermal lens microscope (TLM) 相似文献
16.
Pedrón I Chisvert A March JG Salvador A Benedé JL 《Analytical and bioanalytical chemistry》2011,400(2):595-601
A novel and simple device for membrane-assisted liquid-phase microextraction is used for the first time in a three-phase system.
The device consists of a glass vial containing the aqueous acceptor phase, whose septum of its screw stopper has been replaced
by a sized piece of polytetrafluoroethylene membrane impregnated with n-decane. The vial is assembled to a volumetric flask containing the aqueous donor phase, and the membrane comes in contact
alternatively with both donor and acceptor aqueous phases by orbital agitation. The device has been tested for the determination
of nitrite in tap water samples, which is extensively carried out in routine analysis, as model analytical application. Experimental
variables, such as the organic solvent used to form the supported liquid membrane, the volumes of both donor and acceptor
phases, the orbital agitation rate, and the extraction time were studied and optimized in terms of enrichment factor. Under
the selected working conditions, the analytical figures of merit for nitrite determination were a linearity range up to 50 ng mL−1, limits of detection and quantification of 0.15 and 0.50 ng mL−1, respectively, and a good repeatability (RSD < 10%). The method has been applied to four tap water samples of different origins,
and accurate and precise results were achieved. Besides, the very low volume of organic solvent used, its low cost and the
no-risk of cross-contamination are significant operational advantages. 相似文献
17.
A potentiometric azide-selective sensor based on the use of iron(III) hydrotris(3,5-dimethylpyrazolyl)borate acetylacetonate
chloride [TpMe2Fe(acac)Cl] as a neutral carrier for an azide-selective electrode is reported. Effect of various plasticizers, viz. o-nitrophenyloctyl ether (o-NPOE), dioctylphthalate (DOP), dibutylphthalate (DBP), and benzylacetate (BA), and an anion excluder, hexadecyltrimethylammonium
bromide (HTAB), with [TpMe2Fe(acac)Cl] complex in poly(vinyl chloride) (PVC) were studied. The best performance was obtained with a membrane composition
of [TpMe2Fe(acac)Cl]/HTAB/DOP/PVC in a ratio of 5:2:190:100 (w/w). The sensor exhibits significantly enhanced selectivity toward azide
ions over the concentration range 6.3 × 10−7 to 1.0 × 10−2 M with a lower detection limit of 3.8 × 10–7 M and a Nernstian slope of 59.4 ± 1.1 mV decade−1. Influences of the membrane composition, pH and possible interfering anions were investigated on the response properties
of the electrode. Fast and stable response, good reproducibility, long-term stability and applicability over a wide pH range
(3.5–9.0) are demonstrated. The sensor has a response time of 14 s and can be used for at least 45 days without any considerable
divergence in the potential response. The proposed electrode shows fairly good discrimination of azide from several inorganic
and organic anions. It was successfully applied to the direct determination of azide in orange juice, tea extracts and human
urine samples. 相似文献
18.
Microwave digestion and isotope dilution inductively coupled plasma mass spectrometry (ID-ICP-SFMS) has been applied to the
determination of Pb in rice flour. In order to achieve highly precise determination of low concentrations of Pb, the digestion
blank for Pb was reduced to 0.21 ng g−1 after optimization of the digestion conditions, in which 20 mL analysis solution was obtained after digestion of 0.5 g rice
flour. The observed value of Pb in a non-fat milk powder certified reference material (CRM), NIST SRM 1549, was 16.8 ± 0.8 ng
g−1 (mean ± expanded uncertainty, k = 2; n = 5), which agreed with the certified value of 19 ± 3 ng g−1 and indicated the effectiveness of the method. Analytical results for Pb in three brown rice flour CRMs, NIST SRM 1568a,
NIES CRM 10-a, and NIES CRM 10-b, were 7.32 ± 0.24 ng g−1 (n = 5), 1010 ± 10 ng g−1 (n = 5), and 1250 ± 20 ng g−1 (n = 5), respectively. The concentration of Pb in a candidate white rice flour reference material (RM) sample prepared by the
National Metrology Institute of Japan (NMIJ) was observed to be 4.36 ± 0.28 ng g−1 (n = 10 bottles).
Figure Digestion blank of Pb was carefully reduced to approximately 0.2 ng g-1 which permitted the highly precise determination of Pb at low ng g-1 level in foodstuff samples by ID-SFMS 相似文献
19.
Serra B Gamella M Reviejo AJ Pingarrón JM 《Analytical and bioanalytical chemistry》2008,391(5):1853-1860
The use of lectins for microorganism biosensors fabrication is proposed. Lectins are immobilised onto a gold-plated quartz
crystal for direct piezoelectric label-free transduction of the bacteria–lectin binding event using an electrochemical quartz
crystal microbalance (EQCM). Concanavalin A (Con A) and Escherichia coli were used for the evaluation of the lectin immobilisation method and the biosensor performance. Adsorption on nonpolarised
and polarised (−0.200 V) gold-coated quartz crystals and immobilisation through avidin–biotin binding were checked for Con
A surface attachment. Lectin–bacteria binding was evaluated in all cases. With a crystal modified with Con A via avidin–biotin
immobilisation we obtained a linear calibration plot between 5.0 × 106 and 2.0 × 107 cfu mL−1 by measuring frequency changes with E. coli concentration 1 h after bacteria addition. A remarkable increase in sensitivity was achieved when the analytical solution
contained free biotinylated Con A, as a consequence of multiple lectin adhesion to Escherichia coli cell wall, which produced an accumulation of Con A–E. coli conjugates in the form of multilayers at the electrode surface. A detection limit of approximately 1.0 × 104 cfu mL−1 was achieved. Moreover nonspecific adsorptions were minimised. Using Con A and lectin from Arachis hypogaea, different response profiles were achieved for Escherichia coli, Staphylococcus aureus and Mycobacterium phlei, thus demonstrating the feasibility of bacteria discrimination. An approach involving filtering of free and lectin-bound
bacteria and introduction of a filter in the measuring cell allowed a significant frequency change to be obtained for an E. coli concentration of 1.0 × 103 cfu mL−1 in order to further increase the sensitivity and discriminate between viable and nonviable cells; an approach using electrochemical
measurements of bacterial catalase activity was also checked. 相似文献
20.
A new H2O2 biosensor was fabricated on the basis of nanocomposite films of hemoglobin (Hb), silver nanoparticles (AgNPs), and multiwalled
carbon nanotubes (MWNTs)–chitosan (Chit) dispersed solution immobilized on glassy carbon electrode (GCE). The immobilized
Hb displayed a pair of well-defined and reversible redox peaks with a formal potential (E
θ′) of −22.5 mV in 0.1 M pH 7.0 phosphate buffer solution. The apparent heterogeneous electron transfer rate constants (k
s) in the Chit–MWNTs film was evaluated as 2.58 s−1 according to Laviron’s equation. The surface concentration (Γ*) of the electroactive Hb in the Chit–MWNTs film was estimated to be (2.48 ± 0.25) × 10−9 mol cm−2. Meanwhile, the Chit–MWNTs/Hb/AgNPs/GCE demonstrated excellently electrocatalytical ability to H2O2. Its apparent Michaelis–Menten constant (K
Mapp) for H2O2 was 0.0032 mM, showing a good affinity. Under optimal conditions, the biosensors could be used for the determination of H2O2 ranging from 6.25 × 10−6 to 9.30 × 10−5 mol L−1 with a detection limit of 3.47 × 10−7 mol L−1 (S/N = 3). Furthermore, the biosensor possessed rapid response to H2O2 and good stability, selectivity, and reproducibility. 相似文献