Selectivity and sensitivity of a reagentless electrochemical DNA sensor studied by square wave voltammetry and fluorescence

Poly(5-hydroxy-1,4-naphthoquinone-co-5-hydroxy-3-thioacetic acid-1,4-naphthoquinone)-modified electrode is used for the direct electrochemical detection of oligonucleotide hybridization. The polymer film presents well-defined electroactivity in the cathodic potential domain (between 0 and -0.8 V/SCE), due to the quinone group embedded into the polymer structure. The detection can be performed simply by square wave voltammetry. This sensor is a "signal-on" device and works with different oligonucleotide lengths, from 10 to 30 bases. Quantitative results from fluorescence are consistent with electrochemical data. It is confirmed that the signal increase in square wave voltammetry is unambiguously due to hybridization. The biosensor presents a detection limit of target of ca. 25 nM and is highly selective as it can discriminate single mismatch base.     

Functionalization of single-walled carbon nanotubes for direct and selective electrochemical detection of DNA

We report here a new strategy to graft both redox and DNA probes on carbon nanotubes to make a label-free DNA sensor. Oxidized single-walled carbon nanotubes are first immobilized on a self-assembled monolayer of cysteamine; then the redox probe, a quinone derivative 3-[(2-aminoethyl)sulfanyl-5-hydroxy-1,4-naphthoquinone], is grafted on the free carboxylic groups of the nanotubes. After that, for DNA probe grafting, new carboxylic sites are generated via an aryl diazonium route. After hybridization with a complementary sequence, the conformational changes of DNA could influence the redox kinetics of quinone, leading to a current increase of the redox signal, detected by square wave voltammetry. The system is selective, as it can discriminate a single mismatched sequence from the complementary one.     

Potentiometric determination of glucose by enzymatic oxidation in a flow system

A potentiometric determination is described for glucose based on oxidation by 1,4-benzoquinone with immobilized glucose oxidase as catalyst in an enzyme reactor. The electrode is preceded by an analytical dialysis unit to remove proteins. The ratio of quinone to hydroquinone was measured with a flow-through gold electrode. Another gold electrode preceded the enzyme reactor to correct for serum components (e.g. ascorbic acid) which can also reduce quinone. The operating range is 0.04–10 × 10^-3 M β-D-glucose. The dialysis proceeds with a linear dependence on glucose concentration, and the dialysis ratio can be adjusted by changing the buffer flow rate.     

Design of a new electrogenerated polyquinone film substituted with glutathione. Towards direct electrochemical biosensors

We developed a method to graft a tripeptide (glutathione) onto 5-hydroxy-1,4-naphthoquinone, an electropolymerizable molecule. The resulting thin conducting polymer presents a well-defined and stable electroactivity in neutral buffered solution, due to the embedded quinone group, and is able to covalently graft amino-modified DNA probe strands. It is shown that the bioelectrode presents positive current change following DNA hybridization. This makes a “signal-on” direct electrochemical DNA sensor. The results were obtained with low target concentration (50 nM) and the selectivity is excellent as a single-mismatch sequence can be discriminated from the full-complementary target.     

Detection of Adrenaline in Blood Plasma as Biomarker for Adrenal Venous Sampling

An amperometric bi‐enzyme biosensor based on substrate recycling principle for the amplification of the sensor signal has been developed for the detection of adrenaline in blood. Adrenaline can be used as biomarker verifying successful adrenal venous sampling procedure. The adrenaline biosensor has been realized via modification of a galvanic oxygen sensor with a bi‐enzyme membrane combining a genetically modified laccase and a pyrroloquinoline quinone‐dependent glucose dehydrogenase. The measurement conditions such as pH value and temperature were optimized to enhance the sensor performance. A high sensitivity and a low detection limit of about 0.5–1 nM adrenaline have been achieved in phosphate buffer at pH 7.4, relevant for measurements in blood samples. The sensitivity of the biosensor to other catecholamines such as noradrenaline, dopamine and dobutamine has been studied. Finally, the sensor has been successfully applied for the detection of adrenaline in human blood plasma.     

4-Amino-1-naphthylphosphate as a substrate for the amperometric detection of alkaline phosphatase activity and its application for immunoassay

Immunosensors and biochemical array detection systems based on electrochemical transducers have many advantages such as low detection limit, fast response, simple design and ease of miniaturization. However, further development of such sensors will depend on the availability of suitable substrates that can be converted by a labeling enzyme to an electrochemically active product. Here, we report the synthesis of 4-amino-1-naphthylphosphate and it’s application as a new substrate for alkaline phosphatase. The electrochemical and enzymatic properties of this compound were investigated and compared with the properties of other aromatic 1,4-dihydroxy and 1,4-hydroxy-amine derivatives. The product of the enzyme reaction was 4-aminonaphthol, which was rapidly converted in the presences of air to 1,4-iminonaphthoquinone. This compound could then be detected in an amperometric flow injection assay (AFIA) with −200 mV versus Ag/AgCl potential application. The analytical range for mouse IgG, in an alkaline phosphatase amplified sandwich immuoassay with amperometric detection, was 0.01-100 μg ml⁻¹.     

Direct detection of formaldehyde in air by a novel NAD+- and glutathione-independent formaldehyde dehydrogenase-based biosensor

An amperometric enzyme-based sensor-system for the direct detection of formaldehyde in air is under investigation. The biosensor is based on a native bacterial NAD(+)- and glutathione-independent formaldehyde dehydrogenase as biorecognition element. The enzyme was isolated from Hyphomicrobium zavarzinii strain ZV 580, grown on methylamine hydrochloride in a fed-batch process. The sensor depends on the enzymatic conversion of the analyte to formic acid. Released electrons are detected in an amperometric measurement at 0.2V vs. Ag/AgCl reference electrode by means of a redox-mediator. To optimize the sensing device, Ca(2+) and pyrroloquinoline quinone (PQQ) were added to the buffer solution as reconstitutional substances. At this stage, the sensor shows linear response in the tested ppm-range with a sensitivity of 0.39 microA/ppm. The signal is highly reproducible with respect to sensitivity and base line signal. Reproducibility of sensitivity is more than 90% within the same bacterial batch and even when enzyme of different bacterial batches is used.     

Synthesis of Heterocycles Based on Products of Anion Arylation of Unsaturated Compounds. 5. Reaction of 2-Aryl-1,4-benzoquinones with Thiourea

2-Aryl-1,4-benzoquinones, that can be obtained by arylation of quinone by arenediazonium salts, regioselectively react with thiourea in acid medium to form 7-aryl-5-hydroxy-1,3-benzoxathiol-2-ones. In the presence of excess arylquinones at room temperature, 2-amino-6-hydroxybenzothiazoles are formed in the same reaction.     

First synthesis of N-(3-carboxylpropyl)-5-amino-2-hydroxy-3- tridecyl-1,4-benzoquinone, an unusual quinone isolated from Embelia ribes

The first synthesis of the unusual nitrogen-containing 3-alkyl-1,4-benzoquinone, N-(3-carboxylpropyl)-5-amino-2-hydroxy-3-tridecyl-1,4-benzoquinone, isolated from Embelia ribes, is reported. The key steps are a microwave-assisted combined Mitsunobu reaction-Claisen rearrangement to introduce the alkyl side chain into 2,5-dimethoxyphenol, followed by alkene reduction, oxidation to the quinone, and sequential displacement of the methoxy groups with hydroxide and GABA tert-butyl ester. Two other naturally occurring benzoquinones, O-methylrapanone and rapanone, were also prepared en route.     

Structure of plumbazeylanone: a novel trimer of plumbagin from plumbago zeylanica

Plumbazeylanone, a quinone from Plumbago zeylanica is probably 5b,11a,12,12a-tetrahydro-1,7-dihydroxy-5b-(8-hydroxy-3-methyl-1,4-naphthoquinon-2-yl)-5a,12a-dimethyl-5a$\text{H}$-dibenzo[$\text{b}$,$\text{h}$]fluorene-5,13:6,11-diquinone, a novel trimer of plumbagin with an additional methyl group.     

Temporary inactivation of plasma amine oxidase by alkylhydrazines. A combined enzyme/model study implicates cofactor reduction/reoxidation but cofactor deoxygenation and subsequent reoxygenation in the case of hydrazine itself

It has been known for some time that hydrazine and its methyl and 1,1-dimethyl analogues induce inactivation of the copper-containing quinone-dependent plasma amine oxidase but that the activity recovers over time, suggesting metabolism of all three inhibitors. However, the mechanism responsible for loss and regain of activity has not been investigated. In this study a combination of enzyme studies under a controlled atmosphere along with model studies using 5-tert-butyl-2-hydroxy-1,4-benzoquinone to mimic the 2,4,5-trihydroxyphenylalanine quinone (TPQ) cofactor of the enzyme suggest that regain of enzyme activity represents two different O(2)-dependent processes. In the case of methylhydrazine and 1,1-dimethylhydrazine, we propose that the inactive methylhydrazone/azo form of the enzyme slowly rehydrates and eliminates MeN=NH to give the triol cofactor form, which instantly reoxidizes to the catalytically active quinone form in the presence of O(2). Metabolism of methylhydrazine represents its conversion to CH(4) and N(2), and of 1,1-dimethylhydrazine to CH(2)=O, CH(4), and N(2). In the case of hydrazine itself, however, we propose that the inactive hydrazone/azo form of the enzyme instead undergoes a slow decomposition, probably facilitated by the active-site copper, to give N(2) and a novel 5-desoxy resorcinol form of the cofactor. The latter undergoes a rapid, but noninstantaneous reoxygenation at C5 to restore the active cofactor form, also probably mediated by the active-site copper.     

Total synthesis of maesanin and analogues

An efficient total synthesis of maesanin and related quinones is reported, through direct alkylation of 1,2,4,5-tetramethoxybenzene with alkylbromides, followed by oxidation with ceric ammonium nitrate (CAN) which provokes formation of the quinone and deprotection of the more hindered methyl ether in one step, to furnish the desired 2-hydroxy-5-methoxy-1,4-benzoquinones 1a-h.     

Amperometric Detection of Catecholamine Neurotransmitters Using Electrocatalytic Substrate Recycling at a Laccase Electrode

An enzyme electrode based on the coimmobilization of an osmium redox polymer and laccase on glassy carbon electrodes has been applied to ultra sensitive amperometric detection of the catecholamine neurotransmitters dopamine, epinephrine and norepinephrine, resulting in nanomolar detection limits, as low as 4 nM for dopamine. The sensitivity of the electrode is due to signal amplification via oxidation of the catecholamine by the immobilized laccase, which is regenerated by concomitant reduction of oxygen to water, coupled to the electrocatalytic re‐reduction of the oxidized catecholamine by the osmium redox complex: electrocatalytic substrate recycling. In addition because the sensor can be operated in reductive mode at ?0.2 V (vs. Ag/AgCl), noise and interferences are diminished. Combined with its high sensitivity this enzyme electrode also exhibited excellent selectivity allowing the detection of catecholamines in the presence of ascorbic acid. However, differentiation between the current responses achieved for the three catecholamines is not possible. The effective mode of constant recycling, resulting in amplification of the current response, of the laccase enzyme electrode sensor combined with the inherent advantages of using electrochemical techniques holds great promise for the future of catecholamine detection and monitoring.     

Photochemistry of methoxyhydroquinone and methoxy-p-benzoquinone in solution related to the photoyellowing of the lignocellulosics.

The photoreactivity of methoxy-p-benzoquinone (MQ) and methoxyhydroquinone (MHQ) in dilute solution (10(-4)-10(-3) M) was studied using continuous irradiation and laser flash photolysis (LFP). The quinone irradiated in degassed tetrahydrofuran (THF) gives MHQ and an adduct with the solvent. Only the formation of hydroquinone is observed in ethanol, and hydroxylation is evidenced in water, whereas the compound is stable in CCl4. The bis-quinone, 4,4'-dimethoxybiphenyl-2,5,2',5'-bisquinone, and the dibenzofurane-quinone, 8-hydroxy-3,7-dimethoxydibenzofuran-1,4-quinone, are formed in the presence of MHQ, whereas the reactivity is low with ethylconiferyl alcohol. When MHQ is irradiated selectively in degassed THF, the formation of MQ and of the bis-hydroquinone, 4,4'-dimethoxy-2,5,2',5'-tetrahydroxy-biphenyl, are observed. The dimer is oxidized photochemically or thermally into the mono- or bis-quinones, the process being accelerated in alkaline medium. The formation of the dimers is strongly favored by the contiguous presence of quinone and hydroquinone. When MHQ is selectively irradiated in the presence of transethylconiferaldehyde (EtC), quinone formation and isomerization of EtC are observed. LFP experiments, performed with a selective excitation of MQ, indicate that the triplet state of the quinone is strongly quenched by MHQ to conduce to a semiquinone radical. The interaction between 3MQ* and MQ is mainly driven by an electron transfer process according to the similar value of the quenching rate constant found with another electron donor compound such as 1,4-dimethoxybenzene. By contrast, no strong quenching of 3EtC* by MHQ was observed. It is proposed that the photochemistry of the couple MQ/MHQ is governed by the formation of encounter complex between either 3MQ* and MHQ or 3MHQ* and MQ. Consequently, the fast part of the photoyellowing of lignocellulosics does not appear to involve the couple MHQ/MQ or MHQ/etherified coniferaldehyde, but more likely a combination of oxidation of the hydroquinone by ground-state oxygen and photohydration of the formed quinone from its triplet state, giving inter-alia more colored o-quinonoid type molecules.     

1,4-Carbazolchinone ausp-Benzochinon und primären aliphatischen Aminen. Das vermeintliche und das wahre 2-Methylamino-benzochinon

The compound which is obtained in the reaction of 1,4-benzoquinone with methylamine in chloroform was supposed to be 2-methylaminobenzoquinone; actually it is 6-hydroxy-9-methyl-3-methylamino-1,4-carbazolquinone. The structure is proved by spectroscopic methods, through derivates, zinc dust distillation and via synthesis from 2,5,2,5-biphenyl-diquinone. A second main product, 2,5-bis(methylamino)quinone, is obtained; varying the reaction conditions shows especially the influence of the solvent. One of the intermediate products is the true 2-methylamino-1,4-quinone. Other substituted 9-alkyl-3-alkylamino-6-hydroxy-1,4-carbazolquinones can be prepared by reaction with the corresponding primary amines.

     

Role of LHCII organization in the interaction of substituted 1,4-anthraquinones with thylakoid membranes

The chlorophyll fluorescence, photochemical activity and surface electric properties of thylakoid membranes with different stoichiometry of pigment-protein complexes and organization of the light-harvesting chlorophyll a/b protein complex of photosystem II (LHCII) were studied in the presence of substituted 1,4-anthraquinones. Data show strong dependence of the quenching of the chlorophyll fluorescence on the structural organization of LHCII. The increase of the LHCII oligomerization, which is associated with significant reduction of the transmembrane electric charge asymmetry and electric polarizability of the membrane, correlates with enhanced quenching effect of substituted 1,4-athraquinones. Crucial for the large quinone-induced changes in the membrane electric dipole moments is the structure of the quinone molecule. The strongest reduction in the values of the dipole moments is observed after interaction of thylakoids with 3-chloro-9-hydroxy-1,4-anthraquinone (TF33) which has the highest quenching efficiency. The quinone induced changes in the photochemical activity of photosystem II (PSII) correlate with the total amount of the supramolecular LHCII-PSII complex and depend on the number of substituents in the 1,4-anthraquinone molecule.     

Flow-injection determination of l-ascorbate with cucumber juice as carrier

Cucumber (Cucumis sativus L.) juice has a high l-ascorbate oxidase activity and can be used as the carrier solution in an amperometric flow-injection system for the determination of l-ascorbate. The determination time is 1 min. The calibration graph is linear over the range 5×10^?4–7×10^?3 M (RSD 4%). No enzyme purification is needed. The juice solution retains its activity for 8 days with recycling.     

L-Cysteine Determination Based on Tyrosinase Amperometric Biosensors without Interferences from Thiolic Compounds

This paper presents an enzymatic analysis method for selective detection of L-cysteine (L-Cys) without interferences from other thiols like cysteamine (CA) and mercaptoacetic acid (MAC). The amperometric biosensors are based on tyrosinase (Tyr) that converts catechol to o-quinone. The L-Cys detection is based on the fact that all thiols (including L-Cys) react with o-quinone producing electroinactive compounds and only interfering thiols (CA and MAC) inhibit Tyr. One sample was analyzed twice: with Tyr immobilized on WE surface to quantify enzyme inhibition by thiolic interferents and with Tyr free in solution to investigate the reactions between quinone and all thiolic compounds.     

带有长侧链醌类化合物的Langmuir膜及其表面电势

泛醌(Ubiquinone)和质体醌(Plastoquinone)作为非蛋白电子载体存在于动物线粒体呼吸链和植物光合作用链中.这两类物质在生物膜上有活性.     

2‐(Pyrrolidin‐1‐yl)‐1,4‐naphtho­quinone and 2‐phenyl­sulfanyl‐3‐(pyrrolidin‐1‐yl)‐1,4‐naphtho­quinone

The structure of 2‐(pyrrolidin‐1‐yl)‐1,4‐naphtho­quinone, C₁₄H_12.95Cl_0.05NO₂, (I), is actually a 0.95:0.05 mixture including 2‐chloro‐3‐(pyrrolidin‐1‐yl)‐1,4‐naphtho­quinone as a minor impurity, but (I) was resolved as a single molecule containing a Cl atom with 5% occupancy at the 3‐position. Compound (I) was prepared from the fully chloro‐substituted analogue in an attempt to produce the disubstituted pyrrolidinyl derivative. 2‐Phenyl­sulfanyl‐3‐(pyrrolidin‐1‐yl)‐1,4‐naphtho­quinone, C₂₀H₁₇NO₂S, (II), was also prepared from 2‐chloro‐3‐(pyrrolidin‐1‐yl)‐1,4‐naphtho­quinone, using a strong exocyclic nucleophile. The structure of (II) differs from previous structures of 2,3‐di­chloro‐1,4‐naphtho­quinone and its derivatives in that the naphtho­quinone ring is non‐planar.