World Congress on Biosensors 2014

World Congress on Biosensors 2014
Biosensors 2014

Monday, 7 January 2013

Just Published: Analytica Chimica Acta


A new issue of this journal has just been published. To see abstracts of the papers it contains (with links through to the full papers) click here:
Selected papers from the latest issue:

Current trends in separation of plasmid DNA vaccines: A review

07 January 2013, 10:02:34
14 January 2013
Publication year: 2013
Source:Analytica Chimica Acta, Volume 760

Plasmid DNA (pDNA)-based vaccines offer more rapid avenues for development and production if compared to those of conventional virus-based vaccines. They do not rely on time- or labour-intensive cell culture processes and allow greater flexibility in shipping and storage. Stimulating antibodies and cell-mediated components of the immune system are considered as some of the major advantages associated with the use of pDNA vaccines. This review summarizes the current trends in the purification of pDNA vaccines for practical and analytical applications. Special attention is paid to chromatographic techniques aimed at reducing the steps of final purification, post primary isolation and intermediate recovery, in order to reduce the number of steps necessary to reach a purified end product from the crude plasmid.

Graphical abstract

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Highlights

Strategies for purifying supercoiled plasmid DNA.
► Current trends in the separation and purification of Plasmid DNA vaccines. ► Practical large and analytical scale productions of the plasmid DNA vaccine are discussed. ► Separation challenges. ► New developments and process solutions. ► Future prospective in the field of plasmid DNA vaccines.

Quantitative analysis of the effect of zidovudine, efavirenz, and ritonavir on insulin aggregation by multivariate curve resolution alternating least squares of infrared spectra

07 January 2013, 10:02:34
14 January 2013
Publication year: 2013
Source:Analytica Chimica Acta, Volume 760

Quantification of the effect of antiretroviral drugs on the insulin aggregation process is an important area of research due to the serious metabolic diseases observed in AIDS patients after prolonged treatment with these drugs. In this work, multivariate curve resolution alternating least squares (MCR-ALS) was applied to infrared monitoring of the insulin aggregation process in the presence of three antiretroviral drugs to quantify their effect. To evidence concentration dependence in this process, mixtures at two different insulin:drug molar ratios were used. The interaction between insulin and each drug was analysed by 1H NMR spectroscopy. In all cases, the aggregation process was monitored during 45min by infrared spectroscopy. The aggregates were further characterised by scanning electron microscopy (SEM). MCR-ALS provided the spectral and concentration profiles of the different insulin–drug conformations that are involved in the process. Their feasible band boundaries were calculated using the MCR-BANDS methodology. The kinetic profiles describe the aggregation pathway and the spectral profiles characterise the conformations involved. The retrieved results show that each of the three drugs modifies insulin conformation in a different way, promoting the formation of aggregates. Ritonavir shows the strongest promotion of aggregation, followed by efavirenz and zidovudine. In the studied concentration range, concentration dependence was only observed for zidovudine, with shorter aggregation time obtained as the amount of zidovudine increased. This factor also affected the aggregation pathway.

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Highlights

► The structure of insulin can be changed via interaction with antiretroviral drugs. ► The chemical interaction promotes the formation of aggregates. ► This drug effect was evaluated by MCR-ALS coupled to IR spectroscopy. ► Formation of aggregates was favourable if drugs were able to form hydrogen bonds. ► Higher drug concentrations favoured formation of amorphous aggregates.

Sample size planning for classification models

07 January 2013, 10:02:34
14 January 2013
Publication year: 2013
Source:Analytica Chimica Acta, Volume 760

In biospectroscopy, suitably annotated and statistically independent samples (e.g. patients, batches, etc.) for classifier training and testing are scarce and costly. Learning curves show the model performance as function of the training sample size and can help to determine the sample size needed to train good classifiers. However, building a good model is actually not enough: the performance must also be proven. We discuss learning curves for typical small sample size situations with 5–25 independent samples per class. Although the classification models achieve acceptable performance, the learning curve can be completely masked by the random testing uncertainty due to the equally limited test sample size. In consequence, we determine test sample sizes necessary to achieve reasonable precision in the validation and find that 75–100 samples will usually be needed to test a good but not perfect classifier. Such a data set will then allow refined sample size planning on the basis of the achieved performance. We also demonstrate how to calculate necessary sample sizes in order to show the superiority of one classifier over another: this often requires hundreds of statistically independent test samples or is even theoretically impossible. We demonstrate our findings with a data set of ca. 2550 Raman spectra of single cells (five classes: erythrocytes, leukocytes and three tumour cell lines BT-20, MCF-7 and OCI-AML3) as well as by an extensive simulation that allows precise determination of the actual performance of the models in question.

Graphical abstract

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Highlights

► We compare sample size requirements for classifier training and testing. ► Number of training samples: determine from learning curve. ► Test sample size: specify confidence interval width or model to compare to. ► Classifier testing needs far more samples than training. ► Start with at least 75 cases per class, then refine sample size planning.

Predictive-property-ranked variable reduction in partial least squares modelling with final complexity adapted models: Comparison of properties for ranking

07 January 2013, 10:02:34
14 January 2013
Publication year: 2013
Source:Analytica Chimica Acta, Volume 760

The calibration performance of partial least squares regression for one response (PLS1) can be improved by eliminating uninformative variables. Many variable-reduction methods are based on so-called predictor-variable properties or predictive properties, which are functions of various PLS-model parameters, and which may change during the steps of the variable-reduction process. Recently, a new predictive-property-ranked variable reduction method with final complexity adapted models, denoted as PPRVR-FCAM or simply FCAM, was introduced. It is a backward variable elimination method applied on the predictive-property-ranked variables. The variable number is first reduced, with constant PLS1 model complexity A, until A variables remain, followed by a further decrease in PLS complexity, allowing the final selection of small numbers of variables. In this study for three data sets the utility and effectiveness of six individual and nine combined predictor-variable properties are investigated, when used in the FCAM method. The individual properties include the absolute value of the PLS1 regression coefficient (REG), the significance of the PLS1 regression coefficient (SIG), the norm of the loading weight (NLW) vector, the variable importance in the projection (VIP), the selectivity ratio (SR), and the squared correlation coefficient of a predictor variable with the response y (COR). The selective and predictive performances of the models resulting from the use of these properties are statistically compared using the one-tailed Wilcoxon signed rank test. The results indicate that the models, resulting from variable reduction with the FCAM method, using individual or combined properties, have similar or better predictive abilities than the full spectrum models. After mean-centring of the data, REG and SIG, provide low numbers of informative variables, with a meaning relevant to the response, and lower than the other individual properties, while the predictive abilities are similar or better. SIG has the best selective ability of all individual and combined properties, while the predictive ability is similar. REG is faster than SIG. This means that variable reduction with the FCAM method is preferably conducted with properties REG or SIG. The selective ability of REG can be improved by combining it with NLW or VIP.

Graphical abstract

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Highlights

Selected variables after variable reduction by the PPRVR-FCAM method, using individual and combined predictor-variable properties.
► Variable reduction using the PPRVR-FCAM method is investigated. ► Performance of individual and combined predictor-variable properties is studied. ► Selective and predictive performances of resulting models statistically compared. ► Absolute PLS1 regression coefficient and its significance are most effective.

Methodology for the validation of analytical methods involved in uniformity of dosage units tests

07 January 2013, 10:02:34
14 January 2013
Publication year: 2013
Source:Analytica Chimica Acta, Volume 760

Validation of analytical methods is required prior to their routine use. In addition, the current implementation of the Quality by Design (QbD) framework in the pharmaceutical industries aims at improving the quality of the end products starting from its early design stage. However, no regulatory guideline or none of the published methodologies to assess method validation propose decision methodologies that effectively take into account the final purpose of developed analytical methods. In this work a solution is proposed for the specific case of validating analytical methods involved in the assessment of the content uniformity or uniformity of dosage units of a batch of pharmaceutical drug products as proposed in the European or US pharmacopoeias. This methodology uses statistical tolerance intervals as decision tools. Moreover it adequately defines the Analytical Target Profile of analytical methods in order to obtain analytical methods that allow to make correct decisions about Content uniformity or uniformity of dosage units with high probability. The applicability of the proposed methodology is further illustrated using an HPLC-UV assay as well as a near infra-red spectrophotometric method.

Graphical abstract

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Highlights

► Methodology to validate methods for uniformity of dosage units tests. ► Valid methods will ensure to make the correct decisions with high probability. ► A Quality by Design compliant validation methodology for UDU assays. ► Analytical Target Profile is defined for UDU assays. ► Application to the validation of an HPLC-UV and NIRS method.

Standard addition method applied to the urinary quantification of nicotine in the presence of cotinine and anabasine using surface enhanced Raman spectroscopy and multivariate curve resolution

07 January 2013, 10:02:34
14 January 2013
Publication year: 2013
Source:Analytica Chimica Acta, Volume 760

In this work, urinary nicotine was determined in the presence of the metabolite cotinine and the alkaloid anabasine using surface enhanced Raman spectroscopy and colloidal gold as substrate. Spectra were decomposed using the multivariate curve resolution-alternating least squares method, and pure contributions were recovered. The standard addition method was applied by spiking urine samples with known amounts of the analyte and relative responses from curve resolution were employed to build the analytical curves. The use of multivariate curve resolution in conjunction with standard addition method showed to be an effective strategy that minimized the need for reagent and time-consuming procedures. The determination of the alkaloid nicotine was successfully accomplished at concentrations 0.10, 0.20 and 0.30μgmL−1 and total error values less than 10% were obtained.

Graphical abstract

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Highlights

► Determination of urinary nicotine in the presence of cotinine and anabasine. ► Surface enhanced Raman spectroscopy for analysis of nicotine. ► Multivariate curve resolution in conjunction with standard addition method. ► Determination of nicotine was accomplished with error values less than 10%.

An absorbing microwave micro-solid-phase extraction device used in non-polar solvent microwave-assisted extraction for the determination of organophosphorus pesticides

07 January 2013, 10:02:34
14 January 2013
Publication year: 2013
Source:Analytica Chimica Acta, Volume 760

A single-step extraction-cleanup method, including microwave-assisted extraction (MAE) and micro-solid-phase extraction (μ-SPE), was developed for the extraction of ten organophosphorus pesticides in vegetable and fruit samples. Without adding any polar solvent, only one kind of non-polar solvent (hexane) was used as extraction solvent in the whole extraction step. Absorbing microwave μ-SPE device, was prepared by packing activated carbon with microporous polypropylene membrane envelope, and used as not only the sorbent in μ-SPE, but also the microwave absorption medium. Some experimental parameters effecting on extraction efficiency was investigated and optimized. 1.0g of sample, 8mL of hexane and three absorbing microwave μ-SPE devices were added in the microwave extraction vessel, the extraction was carried out under 400W irradiation power at 60°C for 10min. The extracts obtained by MAE-μ-SPE were directly analyzed by GC–MS without any clean-up process. The recoveries were in the range of 93.5–104.6%, and the relative standard deviations were lower than 8.7%.

Graphical abstract

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Highlights

► An absorbing microwave μ-SPE device packed with activated carbon was used. ► Absorbing microwave μ-SPE device was made and used to enrich the analytes. ► Absorbing microwave μ-SPE device was made and used to heat samples directly. ► MAE-μ-SPE was applied to the extraction of OPPs with non-polar solvent only.

Dumbbell probe-mediated cascade isothermal amplification: A novel strategy for label-free detection of microRNAs and its application to real sample assay

07 January 2013, 10:02:34
14 January 2013
Publication year: 2013
Source:Analytica Chimica Acta, Volume 760

Considering the great significance of microRNAs (miRNAs) in cancer detection and typing, the development of sensitive, specific, quantitative, and low-cost methods for the assay of expression levels of miRNAs is desirable. We describe a highly efficient amplification platform for ultrasensitive analysis of miRNA (taking let-7a miRNA as a model analyte) based on a dumbbell probe-mediated cascade isothermal amplification (DP-CIA) strategy. The method relies on the circularization of dumbbell probe by binding target miRNA, followed by rolling circle amplification (RCA) reaction and an autonomous DNA machine performed by nicking/polymerization/displacement cycles that continuously produces single-stranded G-quadruplex to assemble with hemin to generate a color signal. In terms of the high sensitivity (as low as 1zmol), wide dynamic range (covering 9 orders of magnitude), good specificity (even single-base difference) and easy operation (one probe and three enzymes), the proposed label-free assay is successfully applied to direct detection of let-7a miRNA in real sample (total RNA extracted from human lung tissue), demonstrating an attractive alternative for miRNA analysis for gene expression profiling and molecular diagnostics, particularly for early cancer diagnosis.

Graphical abstract

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Highlights

► This assay relies on the circularization of dumbbell probe by target microRNA. ► Rolling circle amplification and autonomous DNA machine are then occurred. ► G-quadruplex is continuously produced to bind hemin to generate color signal. ► High sensitivity, wide dynamic range, and good specificity is achieved.

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