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papers from the latest issue:
Utilization of nanoparticle labels for signal amplification in ultrasensitive electrochemical affinity biosensors: A review
23 September 2013,
08:39:12
Publication date: 3 October
2013
Source:Analytica Chimica Acta, Volume 797
Author(s): Liang Ding , Alan M. Bond , Jianping Zhai , Jie Zhang
Nanoparticles with desirable properties not exhibited by the bulk material can be readily synthesized because of rapid technological developments in the fields of materials science and nanotechnology. In particular their highly attractive electrochemical properties and electrocatalytic activity have facilitated achievement of the high level of signal amplification needed for the development of ultrasensitive electrochemical affinity biosensors for the detection of proteins and DNA. This review article explains the basic principles of nanoparticle based electrochemical biosensors, highlights the recent advances in the development of nanoparticle based signal amplification strategies, and provides a critical assessment of the likely drawbacks associated with each strategy. Finally, future perspectives for achieving advanced signal simplification in nanoparticles based biosensors are considered.
Source:Analytica Chimica Acta, Volume 797
Author(s): Liang Ding , Alan M. Bond , Jianping Zhai , Jie Zhang
Nanoparticles with desirable properties not exhibited by the bulk material can be readily synthesized because of rapid technological developments in the fields of materials science and nanotechnology. In particular their highly attractive electrochemical properties and electrocatalytic activity have facilitated achievement of the high level of signal amplification needed for the development of ultrasensitive electrochemical affinity biosensors for the detection of proteins and DNA. This review article explains the basic principles of nanoparticle based electrochemical biosensors, highlights the recent advances in the development of nanoparticle based signal amplification strategies, and provides a critical assessment of the likely drawbacks associated with each strategy. Finally, future perspectives for achieving advanced signal simplification in nanoparticles based biosensors are considered.
Graphical abstract
Quantitative structure–retention relationships models for prediction of high performance liquid chromatography retention time of small molecules: Endogenous metabolites and banned compounds
23 September 2013,
08:39:12
Publication date: 3 October
2013
Source:Analytica Chimica Acta, Volume 797
Author(s): Krzysztof Goryński , Barbara Bojko , Alicja Nowaczyk , Adam Buciński , Janusz Pawliszyn , Roman Kaliszan
Quantitative structure–retention relationship (QSRR) is a technique capable of improving the identification of analytes by predicting their retention time on a liquid chromatography column (LC) and/or their properties. This approach is particularly useful when LC is coupled with a high-resolution mass spectrometry (HRMS) platform. The main aim of the present study was to develop and describe appropriate QSRR models that provide usable predictive capability, allowing false positive identification to be removed during the interpretation of metabolomics data, while additionally increasing confidence of experimental results in doping control area. For this purpose, a dataset consisting of 146 drugs, metabolites and banned compounds from World Anti-Doping Agency (WADA) lists, was used. A QSRR study was carried out separately on high quality retention data determined by reversed-phase (RP-LC–HRMS) and hydrophilic interaction chromatography (HILIC-LC–HRMS) systems, employing a single protocol for each system. Multiple linear regression (MLR) was applied to construct the linear QSRR models based on a variety of theoretical molecular descriptors. The regression equations included a set of three descriptors for each model: ALogP, BELe6, R2p and ALogP2, FDI, BLTA96, were used in the analysis of reversed-phase and HILIC column models, respectively. Statistically significant QSRR models (squared correlation coefficient for model fitting, R 2 =0.95 for RP and R 2 =0.84 for HILIC) indicate a strong correlation between retention time and the molecular descriptors. An evaluation of the best correlation models, performed by validation of each model using three tests (leave-one-out, leave-many-out, external tests), demonstrated the reliability of the models. This paper provides a practical and effective method for analytical chemists working with LC/HRMS platforms to improve predictive confidence of studies that seek to identify small molecules.
Source:Analytica Chimica Acta, Volume 797
Author(s): Krzysztof Goryński , Barbara Bojko , Alicja Nowaczyk , Adam Buciński , Janusz Pawliszyn , Roman Kaliszan
Quantitative structure–retention relationship (QSRR) is a technique capable of improving the identification of analytes by predicting their retention time on a liquid chromatography column (LC) and/or their properties. This approach is particularly useful when LC is coupled with a high-resolution mass spectrometry (HRMS) platform. The main aim of the present study was to develop and describe appropriate QSRR models that provide usable predictive capability, allowing false positive identification to be removed during the interpretation of metabolomics data, while additionally increasing confidence of experimental results in doping control area. For this purpose, a dataset consisting of 146 drugs, metabolites and banned compounds from World Anti-Doping Agency (WADA) lists, was used. A QSRR study was carried out separately on high quality retention data determined by reversed-phase (RP-LC–HRMS) and hydrophilic interaction chromatography (HILIC-LC–HRMS) systems, employing a single protocol for each system. Multiple linear regression (MLR) was applied to construct the linear QSRR models based on a variety of theoretical molecular descriptors. The regression equations included a set of three descriptors for each model: ALogP, BELe6, R2p and ALogP2, FDI, BLTA96, were used in the analysis of reversed-phase and HILIC column models, respectively. Statistically significant QSRR models (squared correlation coefficient for model fitting, R 2 =0.95 for RP and R 2 =0.84 for HILIC) indicate a strong correlation between retention time and the molecular descriptors. An evaluation of the best correlation models, performed by validation of each model using three tests (leave-one-out, leave-many-out, external tests), demonstrated the reliability of the models. This paper provides a practical and effective method for analytical chemists working with LC/HRMS platforms to improve predictive confidence of studies that seek to identify small molecules.
Graphical abstract
Information fusion via constrained principal component regression for robust quantification with incomplete calibrations
23 September 2013,
08:39:12
Publication date: 3 October
2013
Source:Analytica Chimica Acta, Volume 797
Author(s): Frank Vogt
Incomplete calibrations are encountered in many applications and hamper chemometric data analyses. Such situations arise when target analytes are embedded in a chemically complex matrix from which calibration concentrations cannot be determined with reasonable efforts. In other cases, the samples’ chemical composition may fluctuate in an unpredictable way and thus cannot be comprehensively covered by calibration samples. The reason for calibration model to fail is the regression principle itself which seeks to explain measured data optimally in terms of the (potentially incomplete) calibration model but does not consider chemical meaningfulness. This study presents a novel chemometric approach which is based on experimentally feasible calibrations, i.e. concentration series of the target analytes outside the chemical matrix (‘ex situ calibration’). The inherent lack-of-information is then compensated by incorporating additional knowledge in form of regression constraints. Any outside knowledge can be utilized such as literature values of concentration ranges, concentration ratios implied e.g. by stoichiometry, sum parameters to which multiple analytes need to amount to, and/or reasonable signal reconstructions. The core idea is to mitigate the regression principle's strive for the best possible explanation of measured signals toward the best possible explanation under the condition of chemical meaningfulness. As proof-of-principle application, quantitative analyses of selected compounds in microalgae cells have been chosen. After acquiring FTIR calibration spectra from concentration series of 28 analytes, an ex situ calibration model has been built via principal component regression (PCR). Since microalgae biomass is a very complex matrix, the prediction step based on such an incomplete calibration fails. However, after incorporating several regression constraints into PCR predictions, chemically impossible results are avoided as depicted in the graphical abstract. Equally important are enhancements in concentration reproducibility. For most samples in the chosen application, the errorbars were reduced by one order of magnitude. By means of this novel chemometric method, quantitative analyses have been improved so much that cell responses to chemical shifts in their culturing environment can be studied.
Source:Analytica Chimica Acta, Volume 797
Author(s): Frank Vogt
Incomplete calibrations are encountered in many applications and hamper chemometric data analyses. Such situations arise when target analytes are embedded in a chemically complex matrix from which calibration concentrations cannot be determined with reasonable efforts. In other cases, the samples’ chemical composition may fluctuate in an unpredictable way and thus cannot be comprehensively covered by calibration samples. The reason for calibration model to fail is the regression principle itself which seeks to explain measured data optimally in terms of the (potentially incomplete) calibration model but does not consider chemical meaningfulness. This study presents a novel chemometric approach which is based on experimentally feasible calibrations, i.e. concentration series of the target analytes outside the chemical matrix (‘ex situ calibration’). The inherent lack-of-information is then compensated by incorporating additional knowledge in form of regression constraints. Any outside knowledge can be utilized such as literature values of concentration ranges, concentration ratios implied e.g. by stoichiometry, sum parameters to which multiple analytes need to amount to, and/or reasonable signal reconstructions. The core idea is to mitigate the regression principle's strive for the best possible explanation of measured signals toward the best possible explanation under the condition of chemical meaningfulness. As proof-of-principle application, quantitative analyses of selected compounds in microalgae cells have been chosen. After acquiring FTIR calibration spectra from concentration series of 28 analytes, an ex situ calibration model has been built via principal component regression (PCR). Since microalgae biomass is a very complex matrix, the prediction step based on such an incomplete calibration fails. However, after incorporating several regression constraints into PCR predictions, chemically impossible results are avoided as depicted in the graphical abstract. Equally important are enhancements in concentration reproducibility. For most samples in the chosen application, the errorbars were reduced by one order of magnitude. By means of this novel chemometric method, quantitative analyses have been improved so much that cell responses to chemical shifts in their culturing environment can be studied.
Graphical abstract
Amorphous carbon nitride as an alternative electrode material in electroanalysis: Simultaneous determination of dopamine and ascorbic acid
23 September 2013,
08:39:12
Publication date: 3 October
2013
Source:Analytica Chimica Acta, Volume 797
Author(s): Roberta A. Medeiros , Roberto Matos , Abdelkader Benchikh , Boualem Saidani , Catherine Debiemme-Chouvy , Claude Deslouis , Romeu C. Rocha-Filho , Orlando Fatibello-Filho
Boron-doped diamond (BDD) films are excellent electrode materials, whose electrochemical activity for some analytes can be tuned by controlling their surface termination, most commonly either to predominantly hydrogen or oxygen. This tuning can be accomplished by e.g. suitable cathodic or anodic electrochemical pretreatments. Recently, it has been shown that amorphous carbon nitride (a-CN x ) films may present electrochemical characteristics similar to those of BDD, including the influence of surface termination on their electrochemical activity toward some analytes. In this work, we report for the first time a complete electroanalytical method using an a-CN x electrode. Thus, an a-CN x film deposited on a stainless steel foil by DC magnetron sputtering is proposed as an alternative electrode for the simultaneous determination of dopamine (DA) and ascorbic acid (AA) in synthetic biological samples by square-wave voltammetry. The obtained results are compared with those attained using a BDD electrode. For both electrodes, a same anodic pretreatment in 0.1molL−1 KOH was necessary to attain an adequate and equivalent separation of the DA and AA oxidation potential peaks of about 330mV. The detection limits obtained for the simultaneous determination of these analytes using the a-CN x electrode were 0.0656μmolL−1 for DA and 1.05μmolL−1 for AA, whereas with the BDD electrode these values were 0.283μmolL−1 and 0.968μmolL−1, respectively. Furthermore, the results obtained in the analysis of the analytes in synthetic biological samples were satisfactory, attesting the potential application of the a-CN x electrode in electroanalysis.
Source:Analytica Chimica Acta, Volume 797
Author(s): Roberta A. Medeiros , Roberto Matos , Abdelkader Benchikh , Boualem Saidani , Catherine Debiemme-Chouvy , Claude Deslouis , Romeu C. Rocha-Filho , Orlando Fatibello-Filho
Boron-doped diamond (BDD) films are excellent electrode materials, whose electrochemical activity for some analytes can be tuned by controlling their surface termination, most commonly either to predominantly hydrogen or oxygen. This tuning can be accomplished by e.g. suitable cathodic or anodic electrochemical pretreatments. Recently, it has been shown that amorphous carbon nitride (a-CN x ) films may present electrochemical characteristics similar to those of BDD, including the influence of surface termination on their electrochemical activity toward some analytes. In this work, we report for the first time a complete electroanalytical method using an a-CN x electrode. Thus, an a-CN x film deposited on a stainless steel foil by DC magnetron sputtering is proposed as an alternative electrode for the simultaneous determination of dopamine (DA) and ascorbic acid (AA) in synthetic biological samples by square-wave voltammetry. The obtained results are compared with those attained using a BDD electrode. For both electrodes, a same anodic pretreatment in 0.1molL−1 KOH was necessary to attain an adequate and equivalent separation of the DA and AA oxidation potential peaks of about 330mV. The detection limits obtained for the simultaneous determination of these analytes using the a-CN x electrode were 0.0656μmolL−1 for DA and 1.05μmolL−1 for AA, whereas with the BDD electrode these values were 0.283μmolL−1 and 0.968μmolL−1, respectively. Furthermore, the results obtained in the analysis of the analytes in synthetic biological samples were satisfactory, attesting the potential application of the a-CN x electrode in electroanalysis.
Graphical abstract
Development of hyperbranched polymers with non-covalent interactions for extraction and determination of aflatoxins in cereal samples
23 September 2013,
08:39:12
Publication date: 3 October
2013
Source:Analytica Chimica Acta, Volume 797
Author(s): Xiaoyan Liu , Huihui Li , Zhigang Xu , Jialin Peng , Shuqiang Zhu , Haixia Zhang
A novel approach for assembling homogeneous hyperbranched polymers based on non-covalent interactions with aflatoxins was developed; the polymers were used to evaluate the extraction of aflatoxins B1, B2, G1 and G2 (AFB1, AFB2, AFG1 and AFG2) in simulant solutions. The results showed that the extraction efficiencies of three kinds of synthesized polymers for the investigated analytes were not statistically different; as a consequence, one of the representative polymers (polymer I) was used as the solid-phase extraction (SPE) sorbent to evaluate the influences of various parameters, such as desorption conditions, pH, ionic strength, concentration of methanol in sample solutions, and the mass of the sorbent on the extraction efficiency. In addition, the extraction efficiencies for these aflatoxins were compared between the investigated polymer and the traditional sorbent C18. The results showed that the investigated polymer had superior extraction efficiencies. Subsequently, the proposed polymer for the SPE packing material was employed to enrich and analyze four aflatoxins in the cereal powder samples. The limits of detection (LODs) at a signal-to-noise (S/N) ratio of 3 were in the range of 0.012–0.120ngg−1 for four aflatoxins, and the limits of quantification (LOQs) calculated at S/N=10 were from 0.04 to 0.40ngg−1 for four aflatoxins. The recoveries of four aflatoxins from cereal powder samples were in the range of 82.7–103% with relative standard deviations (RSDs) lower than 10%. The results demonstrate the suitability of the SPE approach for the analysis of trace aflatoxins in cereal powder samples.
Source:Analytica Chimica Acta, Volume 797
Author(s): Xiaoyan Liu , Huihui Li , Zhigang Xu , Jialin Peng , Shuqiang Zhu , Haixia Zhang
A novel approach for assembling homogeneous hyperbranched polymers based on non-covalent interactions with aflatoxins was developed; the polymers were used to evaluate the extraction of aflatoxins B1, B2, G1 and G2 (AFB1, AFB2, AFG1 and AFG2) in simulant solutions. The results showed that the extraction efficiencies of three kinds of synthesized polymers for the investigated analytes were not statistically different; as a consequence, one of the representative polymers (polymer I) was used as the solid-phase extraction (SPE) sorbent to evaluate the influences of various parameters, such as desorption conditions, pH, ionic strength, concentration of methanol in sample solutions, and the mass of the sorbent on the extraction efficiency. In addition, the extraction efficiencies for these aflatoxins were compared between the investigated polymer and the traditional sorbent C18. The results showed that the investigated polymer had superior extraction efficiencies. Subsequently, the proposed polymer for the SPE packing material was employed to enrich and analyze four aflatoxins in the cereal powder samples. The limits of detection (LODs) at a signal-to-noise (S/N) ratio of 3 were in the range of 0.012–0.120ngg−1 for four aflatoxins, and the limits of quantification (LOQs) calculated at S/N=10 were from 0.04 to 0.40ngg−1 for four aflatoxins. The recoveries of four aflatoxins from cereal powder samples were in the range of 82.7–103% with relative standard deviations (RSDs) lower than 10%. The results demonstrate the suitability of the SPE approach for the analysis of trace aflatoxins in cereal powder samples.
Graphical abstract
Hyphenation of optimized microfluidic sample preparation with nano liquid chromatography for faster and greener alkaloid analysis
23 September 2013,
08:39:12
Publication date: 3 October
2013
Source:Analytica Chimica Acta, Volume 797
Author(s): Yao Shen , Teris A. van Beek , Han Zuilhof , Bo Chen
A glass liquid–liquid extraction (LLE) microchip with three parallel 3.5cm long and 100μm wide interconnecting channels was optimized in terms of more environmentally friendly (greener) solvents and extraction efficiency. In addition, the optimized chip was successfully hyphenated with nano-liquid chromatography with ultraviolet and mass spectrometric detection (nanoLC–UV–MS) for on-line analysis. In this system, sample pretreatment, separation and detection are integrated, which significantly shortens the analysis time, saves labor and drastically reduces solvent consumption. Strychnine was used as model analyte to determine the extraction efficiency of the optimized 3-phase chip. Influence of organic solvent, pH of feed phase, type of alkaloid, and flow rates were investigated. The results demonstrated that the 3-phase chip nanoLC–UV/MS hyphenation combines rapid (∼25s) and efficient (extraction efficiency >90%) sample prep, with automated alkaloid analyses. The method was applied to real samples including Strychnos nux-vomica seeds, Cephaelis ipecacuanha roots, Atropa belladonna leaves, and Vinca minor leaves.
Source:Analytica Chimica Acta, Volume 797
Author(s): Yao Shen , Teris A. van Beek , Han Zuilhof , Bo Chen
A glass liquid–liquid extraction (LLE) microchip with three parallel 3.5cm long and 100μm wide interconnecting channels was optimized in terms of more environmentally friendly (greener) solvents and extraction efficiency. In addition, the optimized chip was successfully hyphenated with nano-liquid chromatography with ultraviolet and mass spectrometric detection (nanoLC–UV–MS) for on-line analysis. In this system, sample pretreatment, separation and detection are integrated, which significantly shortens the analysis time, saves labor and drastically reduces solvent consumption. Strychnine was used as model analyte to determine the extraction efficiency of the optimized 3-phase chip. Influence of organic solvent, pH of feed phase, type of alkaloid, and flow rates were investigated. The results demonstrated that the 3-phase chip nanoLC–UV/MS hyphenation combines rapid (∼25s) and efficient (extraction efficiency >90%) sample prep, with automated alkaloid analyses. The method was applied to real samples including Strychnos nux-vomica seeds, Cephaelis ipecacuanha roots, Atropa belladonna leaves, and Vinca minor leaves.
Graphical abstract
Biparametric multicommutated flow analysis system for determination of human serum phosphoesterase activity
23 September 2013,
08:39:12
Publication date: 3 October
2013
Source:Analytica Chimica Acta, Volume 797
Author(s): Łukasz Tymecki , Kamil Strzelak , Robert Koncki
A multicommutated flow analysis (MCFA) system constructed of microsolenoid valves and pumps offering simultaneous determination of activity of acid phosphatase (ACP) and alkaline phosphatase (ALP) in human serum samples has been developed. The MCFA system is based on optoelectronic flow-through detector made of two light emitting diodes and operating according to paired emitter detector diode (PEDD) principle. This photometric PEDD device has been dedicated for detection of p-nitrophenol (NP) generated in the course of enzymatic hydrolysis of p-nitrophenyl phosphate and optimized for the determination of NP in human serum samples. The developed PEDD-based MCFA system allows independent optimization of conditions for reaction and detection steps of photometric ACP and ALP bioassays. Moreover, it allows elimination of photometric interferences from serum matrix components according to two-points kinetic mode of measurement. The single measurement cycle takes 12min, consists of four measurements (two for each phosphoesterase) and enables determination of serum ACP and ALP activities at physiological and pathological levels. The real analytical utility of the developed MCFA system has been confirmed by analysis of control sera as well as real human serum samples from healthy persons and oncological patients.
Source:Analytica Chimica Acta, Volume 797
Author(s): Łukasz Tymecki , Kamil Strzelak , Robert Koncki
A multicommutated flow analysis (MCFA) system constructed of microsolenoid valves and pumps offering simultaneous determination of activity of acid phosphatase (ACP) and alkaline phosphatase (ALP) in human serum samples has been developed. The MCFA system is based on optoelectronic flow-through detector made of two light emitting diodes and operating according to paired emitter detector diode (PEDD) principle. This photometric PEDD device has been dedicated for detection of p-nitrophenol (NP) generated in the course of enzymatic hydrolysis of p-nitrophenyl phosphate and optimized for the determination of NP in human serum samples. The developed PEDD-based MCFA system allows independent optimization of conditions for reaction and detection steps of photometric ACP and ALP bioassays. Moreover, it allows elimination of photometric interferences from serum matrix components according to two-points kinetic mode of measurement. The single measurement cycle takes 12min, consists of four measurements (two for each phosphoesterase) and enables determination of serum ACP and ALP activities at physiological and pathological levels. The real analytical utility of the developed MCFA system has been confirmed by analysis of control sera as well as real human serum samples from healthy persons and oncological patients.
Graphical abstract
Alkyd paints in art: Characterization using integrated mass spectrometry
23 September 2013,
08:39:12
Publication date: 3 October
2013
Source:Analytica Chimica Acta, Volume 797
Author(s): Jacopo La Nasa , Ilaria Degano , Francesca Modugno , Maria Perla Colombini
Alkyd resins have been commonly used as binders in artist paints since the 1940s. The characterization of alkyds in samples from artworks can help to solve attribution and dating issues, investigate decay processes, and contribute to the planning of conservation strategies. Being able to assess the components of industrially formulated paint materials and to differentiate between different trademarks and producers is extremely interesting and requires multi-analytical approaches. In this paper we describe the characterization of commercial alkyd paint materials using a multi-analytical approach based on the integration of three different mass spectrometric techniques: gas chromatography–mass spectrometry (GC/MS), high performance liquid chromatography coupled with electrospray ionization mass spectrometry with a tandem quadrupole-time of flight mass spectrometer (HPLC–ESI-Q-ToF), and flow injection analysis (FIA) in the ESI-Q-ToF mass spectrometer. GC/MS was successful in determining the fatty acid and aromatic fractions of the resins after hydrolysis; HPLC–ESI-Q-ToF analysis enabled us to identify the triglycerides (TAGs) and diglycerides (DAGs) profile of each resin, and FIA analysis was used as a rapid method to evaluate the presence of possible additives such as synthetic polymers.
Source:Analytica Chimica Acta, Volume 797
Author(s): Jacopo La Nasa , Ilaria Degano , Francesca Modugno , Maria Perla Colombini
Alkyd resins have been commonly used as binders in artist paints since the 1940s. The characterization of alkyds in samples from artworks can help to solve attribution and dating issues, investigate decay processes, and contribute to the planning of conservation strategies. Being able to assess the components of industrially formulated paint materials and to differentiate between different trademarks and producers is extremely interesting and requires multi-analytical approaches. In this paper we describe the characterization of commercial alkyd paint materials using a multi-analytical approach based on the integration of three different mass spectrometric techniques: gas chromatography–mass spectrometry (GC/MS), high performance liquid chromatography coupled with electrospray ionization mass spectrometry with a tandem quadrupole-time of flight mass spectrometer (HPLC–ESI-Q-ToF), and flow injection analysis (FIA) in the ESI-Q-ToF mass spectrometer. GC/MS was successful in determining the fatty acid and aromatic fractions of the resins after hydrolysis; HPLC–ESI-Q-ToF analysis enabled us to identify the triglycerides (TAGs) and diglycerides (DAGs) profile of each resin, and FIA analysis was used as a rapid method to evaluate the presence of possible additives such as synthetic polymers.
Graphical abstract
Quantitative analysis of polypeptide antibiotic residues in a variety of food matrices by liquid chromatography coupled to tandem mass spectrometry
23 September 2013,
08:39:12
Publication date: 3 October
2013
Source:Analytica Chimica Acta, Volume 797
Author(s): A. Kaufmann , M. Widmer
A quantitative LC–MS/MS method was developed for the determination of five polypeptide antibiotics (bacitracin, colistin A, colistin B, polymyxin B1 and polymyxin B2) in a variety of food matrices (muscle, liver, kidney, egg and milk). The described method is sufficiently sensitive, selective and provides acceptable recoveries for all compounds. The extraction is based on acidified methanolic solvent. This is followed by a reversed phase solid phase extraction step to clean-up and concentrate the extracts. The use of a modern core shell column in combination with an eluent consisting of trifluoroacetic acid, formic acid and acetonitrile provides chromatographically well resolved analyte peaks The single-step clean-up is fast and produces a sufficiently clean extract in order to control matrix-related signal suppression in the electrospray interface.
Source:Analytica Chimica Acta, Volume 797
Author(s): A. Kaufmann , M. Widmer
A quantitative LC–MS/MS method was developed for the determination of five polypeptide antibiotics (bacitracin, colistin A, colistin B, polymyxin B1 and polymyxin B2) in a variety of food matrices (muscle, liver, kidney, egg and milk). The described method is sufficiently sensitive, selective and provides acceptable recoveries for all compounds. The extraction is based on acidified methanolic solvent. This is followed by a reversed phase solid phase extraction step to clean-up and concentrate the extracts. The use of a modern core shell column in combination with an eluent consisting of trifluoroacetic acid, formic acid and acetonitrile provides chromatographically well resolved analyte peaks The single-step clean-up is fast and produces a sufficiently clean extract in order to control matrix-related signal suppression in the electrospray interface.
Graphical abstract
Sensing of a nucleic acid binding protein via a label-free perylene probe fluorescence recovery assay
23 September 2013,
08:39:12
Publication date: 3 October
2013
Source:Analytica Chimica Acta, Volume 797
Author(s): Dongli Liao , Wenying Li , Jian Chen , Huping Jiao , Huipeng Zhou , Bin Wang , Cong Yu
A novel label-free fluorescence recovery assay for the sensing of a DNA binding protein has been developed. A transcription factor c-Jun protein, and a 21 base pair duplex DNA containing the c-Jun protein binding site (J-DNA) were selected. J-DNA was mixed with a cationic fluorescent perylene probe (compound 1), and induced aggregation of the probe. Quenching of the probe's fluorescence was observed. However, when c-Jun protein was mixed with the J-DNA, c-Jun bound to the duplex DNA, which reduced the degree of the induced perylene probe aggregation, and a turn on fluorescence signal was observed. The recovered fluorescence intensity was directly related to the amount of c-Jun added. The method is highly selective, six non-DNA binding proteins and one randomly selected 21 base pair duplex DNA (con-1) were tested. No noticeable compound 1 fluorescence recovery was observed. Mutations were also introduced to the c-Jun recognition sequence and much reduced fluorescence recovery was observed. Our assay is label-free, convenient, inexpensive, and fast. It can be used in biomedical research such as high throughput screening of drugs targeted at DNA-binding proteins.
Source:Analytica Chimica Acta, Volume 797
Author(s): Dongli Liao , Wenying Li , Jian Chen , Huping Jiao , Huipeng Zhou , Bin Wang , Cong Yu
A novel label-free fluorescence recovery assay for the sensing of a DNA binding protein has been developed. A transcription factor c-Jun protein, and a 21 base pair duplex DNA containing the c-Jun protein binding site (J-DNA) were selected. J-DNA was mixed with a cationic fluorescent perylene probe (compound 1), and induced aggregation of the probe. Quenching of the probe's fluorescence was observed. However, when c-Jun protein was mixed with the J-DNA, c-Jun bound to the duplex DNA, which reduced the degree of the induced perylene probe aggregation, and a turn on fluorescence signal was observed. The recovered fluorescence intensity was directly related to the amount of c-Jun added. The method is highly selective, six non-DNA binding proteins and one randomly selected 21 base pair duplex DNA (con-1) were tested. No noticeable compound 1 fluorescence recovery was observed. Mutations were also introduced to the c-Jun recognition sequence and much reduced fluorescence recovery was observed. Our assay is label-free, convenient, inexpensive, and fast. It can be used in biomedical research such as high throughput screening of drugs targeted at DNA-binding proteins.
Graphical abstract
Cyclically amplified fluorescent detection of theophylline and thiamine pyrophosphate by coupling self-cleaving RNA ribozyme with endonuclease
23 September 2013,
08:39:12
Publication date: 3 October
2013
Source:Analytica Chimica Acta, Volume 797
Author(s): Xuemei Li , Jian Song , Yan Wang , Tao Cheng
A structure-switching-based approach for the design of fluorescent biosensors from known RNA aptazymes were demonstrated for the detection of theophylline and thiamine pyrophosphate (TPP). Taking advantages of the ability of graphene oxide (GO) to protect ssDNA from nuclease cleavage and the cyclic amplification induced by deoxyribonuclease I (DNase I), the amplified assay showed high sensitivity. In the presence of target, the target-dependent hammerhead aptazyme cleaves off. The released Shine–Dalgarno (SD) sequence was introduced into the detection system, in which a FAM labeled probe ssDNA was noncovalently assembled on GO, and the fluorescence of the dye was completely quenched. In the presence of the released sequence, the binding between the dye-labeled DNA and the SD sequence alter the conformation of dye-labeled DNA, and disturb the interaction between the dye-labeled DNA and GO, liberating dye-labeled DNA from GO. The fluorescent intensity was increased, whereupon the DNase I can cleave the free DNA in the DNA/RNA complex, thereby liberating the fluorophore and ultimately releasing the SD RNA sequence. The released SD RNA sequence then binds another DNA probe, and the cycle starts anew, which leads to significant amplification of the fluorescent signal. The strategy showed good sensitivity and the dynamic ranges were of 0.1–10μM and 0.5–100μM for theophylline and TPP, respectively. The approach opens up a wide range of possibilities for sensing of other small molecules in biological entities.
Source:Analytica Chimica Acta, Volume 797
Author(s): Xuemei Li , Jian Song , Yan Wang , Tao Cheng
A structure-switching-based approach for the design of fluorescent biosensors from known RNA aptazymes were demonstrated for the detection of theophylline and thiamine pyrophosphate (TPP). Taking advantages of the ability of graphene oxide (GO) to protect ssDNA from nuclease cleavage and the cyclic amplification induced by deoxyribonuclease I (DNase I), the amplified assay showed high sensitivity. In the presence of target, the target-dependent hammerhead aptazyme cleaves off. The released Shine–Dalgarno (SD) sequence was introduced into the detection system, in which a FAM labeled probe ssDNA was noncovalently assembled on GO, and the fluorescence of the dye was completely quenched. In the presence of the released sequence, the binding between the dye-labeled DNA and the SD sequence alter the conformation of dye-labeled DNA, and disturb the interaction between the dye-labeled DNA and GO, liberating dye-labeled DNA from GO. The fluorescent intensity was increased, whereupon the DNase I can cleave the free DNA in the DNA/RNA complex, thereby liberating the fluorophore and ultimately releasing the SD RNA sequence. The released SD RNA sequence then binds another DNA probe, and the cycle starts anew, which leads to significant amplification of the fluorescent signal. The strategy showed good sensitivity and the dynamic ranges were of 0.1–10μM and 0.5–100μM for theophylline and TPP, respectively. The approach opens up a wide range of possibilities for sensing of other small molecules in biological entities.
Graphical abstract
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