Sample Preparation Approach Research Articles

New Approach for Determining Tortuosity in Fibrous Porous Media

A method to determine tortuosity in a fibrous porous medium is proposed. A new approach for sample preparation and testing has been followed to establish a relationship between air permeability and fiberweb thickness which formed the basis for the determination of tortuosity in fibrous porous media. An empirical relationship between tortuosity and fiberweb structural properties including porosity, fiber diameter and fiberweb thickness has been proposed unlike the models in the literature which have expressed tortuosity as a function of porosity only. Transverse air flow through a fibrous porous media increasingly becomes less tortuous with increasing porosity, with the value of tortuosity approaching 1 at upper limits of porosity. Tortuosity also decreased with increase in fiber diameter whereas increase in fiberweb thickness resulted in the increase in tortuosity within the range of fiberweb thickness tested.

Large-scale pesticide testing in olives by liquid chromatography–electrospray tandem mass spectrometry using two sample preparation methods based on matrix solid-phase dispersion and QuEChERS

In this work we have evaluated the performance of two sample preparation methodologies for the large-scale multiresidue analysis of pesticides in olives using liquid chromatography–electrospray tandem mass spectrometry (LC–MS/MS). The tested sample treatment methodologies were: (1) liquid–liquid partitioning with acetonitrile followed by dispersive solid-phase extraction clean-up using GCB, PSA and C 18 sorbents (QuEChERS method – modified for fatty vegetables) and (2) matrix solid-phase dispersion (MSPD) using aminopropyl as sorbent material and a final clean-up performed in the elution step using Florisil. An LC–MS/MS method covering 104 multiclass pesticides was developed to examine the performance of these two protocols. The separation of the compounds from the olive extracts was achieved using a short C 18 column (50 mm × 4.6 mm i.d.) with 1.8 μm particle size. The identification and confirmation of the compounds was based on retention time matching along with the presence (and ratio) of two typical MRM transitions. Limits of detection obtained were lower than 10 μg kg −1 for 89% analytes using both sample treatment protocols. Recoveries studies performed on olives samples spiked at two concentration levels (10 and 100 μg kg −1) yielded average recoveries in the range 70–120% for most analytes when QuEChERS procedure is employed. When MSPD was the choice for sample extraction, recoveries obtained were in the range 50–70% for most of target compounds. The proposed methods were successfully applied to the analysis of real olives samples, revealing the presence of some of the target species in the μg kg −1 range. Besides the evaluation of the sample preparation approaches, we also discuss the use of advanced software features associated to MRM method development that overcome several limitations and drawbacks associated to MS/MS methods (time segments boundaries, tedious method development/manual scheduling and acquisition limitations). This software feature recently offered by different vendors is based on an algorithm that associates retention time data for each individual MS/MS transition, so that the number of simultaneously traced transitions throughout the entire chromatographic run (dwell times and sensitivity) is maximized.

Influence of Ethanol on Lipid/Sterol Membranes: Phase Diagram Construction from AFM Imaging

Herein we develop a sample preparation approach that enables the use of supported lipid bilayers for the quantitative study of the influence of ethanol (0-20 vol %) on the phase behavior of phospholipid (DPPC)/sterol (ergosterol, 0-20 mol %) bilayers. Three coexisting phases were observed with tapping-mode atomic force microscopy: gel (L(beta)'), liquid-ordered (L(o)), and interdigitated (L(beta)'I). High-resolution imaging permitted the construction of a refined phase diagram for DPPC/ergosterol/ethanol and the observation of L(o)-L(beta)' phase separation that has not been observed using optical techniques. Our results quantitatively show the concentration regime where ergosterol protects the membrane by reducing the membrane fraction that is interdigitated in the presence of ethanol.

Emerging Pesticide Residue Issues and Analytical Approaches†

The 46th Annual Florida Pesticide Residue Workshop of 2009 (FPRW 2009) held in St. Pete Beach, FL, is the latest in an annual tradition drawing scientists from U.S. federal and state government laboratories, industry, and other laboratories worldwide. In 2009, selected FPRW presenters were invited to contribute to this special issue of the Journal of Agricultural and Food Chemistry with a section devoted to emerging pesticide residue issues and analytical approaches. What follows is the written record of what should become a scientific conversation launched at FPRW 2009. There are two distinct approaches to organic residue analysis: instrumental methods and assays. In much of the world, scientists primarily rely on laboratories equipped with instrumentation for analysis, usually gas chromatography and liquid chromatography with some type of selective detector. In the discussion of instrumental approaches, the focus is on chromatography with mass spectrometry as a detection method. Approaches such as biomonitoring and assays fall outside the traditional instrumental method approach to residue analysis. Assays that do not require laboratory equipment are of greater interest for screening and are well-suited to field use. Regardless of the analytical method, the success of multiresidue analysis relies on the appropriate choice of sample preparation and cleanup methodologies. Many new sample preparation and cleanup approaches used for pesticide and other small molecule contaminant residue analyses in a variety of complex sample matrices are discussed in this special issue. The goal of these approaches is to reduce overall analysis time and solvent consumption without compromising the analytical results.

Analysis of 12 beta-lactam antibiotics in human plasma by HPLC with ultraviolet detection

A simple and economical high performance liquid chromatography method was developed and validated for routine analysis of 12 Penicillin, Cephalosporin and Carbapenem antibiotics in 200 μL of human plasma. Antibiotics determined were Ceftazidime, Meropenem, Ceftriaxone, Ampicillin, Cefazolin, Ertapenem, Cephalothin, Benzylpenicillin, Flucloxacillin, Dicloxacillin, Piperacillin and Ticarcillin. There was a common sample preparation approach involving precipitation of proteins with acetonitrile and removal of lipid-soluble components by a chloroform wash. Separations were performed on a Waters X-bridge C18 column with, depending on analytes, one of three acetonitrile–phosphate buffer mobile phases. Detection was by UV at 210, 260 and 304 nm. Validation has demonstrated the method to be linear, accurate and precise. The method has been used in a pathology laboratory for therapeutic drug monitoring (TDM) of beta-lactams in critically ill patients.

In-sample acetylation-non-porous membrane-assisted liquid–liquid extraction for the determination of parabens and triclosan in water samples

A procedure for the determination of seven parabens (esters of 4-hydroxybenzoic acid), including the distinction between branched and linear isomers of propyl- and butyl-parabens and triclosan in water samples, was developed and evaluated. The procedure includes in-sample acetylation-non-porous membrane-assisted liquid-liquid extraction and large volume injection-gas chromatography-ion trap-tandem mass spectrometry. Different derivatisation strategies were considered, i.e. post-extraction silylation with N-methyl-N-(tert-butyldimethylsilyl)-trifluoroacetamide and in situ acylation with acetic anhydride (Ac(2)O) and isobutylchloroformate. Moreover, acceptor solvent and the basic catalyser of the acylation reaction were investigated. Thus, in situ derivatisation with Ac(2)O and potassium hydrogenphosphate (as basic catalyser) was selected. Potassium hydrogenphosphate overcomes some drawbacks of other basic catalysers, e.g. toxicity and bubble formation, while leads to higher responses. Subsequently, other experimental variables affecting derivatisation-extraction yield such as pre-stirring time, salt addition and volume of Ac(2)O were optimised by an experimental design approach. Under optimised conditions, the proposed method achieved detection limits from 0.1 to 1.4 ng L(-1) for a sample volume of 18 mL and extraction efficiencies, estimated by comparison with liquid-liquid extraction, between 46% (for methyl- and ethyl-parabens) and 110% (for benzylparaben). The reported sample preparation approach is free of matrix effects for parabens but affected for triclosan with a reduction of approximately 40% when wastewater samples are analysed; therefore, both internal and external calibration can be used as quantification techniques for parabens, but internal standard calibration is mandatory for triclosan. The application of the method to real samples revealed the presence of these compounds in raw wastewater at concentrations up to 26 ng mL(-1), the prevalence of the linear isomer of propylparaben (n-PrP), and the coexistence of the two isomers of butylparaben (i-BuP and n-BuP) at similar levels.

Gene set enrichment meta-learning analysis: next- generation sequencing versus microarrays

BackgroundReproducibility of results can have a significant impact on the acceptance of new technologies in gene expression analysis. With the recent introduction of the so-called next-generation sequencing (NGS) technology and established microarrays, one is able to choose between two completely different platforms for gene expression measurements. This study introduces a novel methodology for gene-ranking stability analysis that is applied to the evaluation of gene-ranking reproducibility on NGS and microarray data.ResultsThe same data used in a well-known MicroArray Quality Control (MAQC) study was also used in this study to compare ranked lists of genes from MAQC samples A and B, obtained from Affymetrix HG-U133 Plus 2.0 and Roche 454 Genome Sequencer FLX platforms. An initial evaluation, where the percentage of overlapping genes was observed, demonstrates higher reproducibility on microarray data in 10 out of 11 gene-ranking methods. A gene set enrichment analysis shows similar enrichment of top gene sets when NGS is compared with microarrays on a pathway level. Our novel approach demonstrates high accuracy of decision trees when used for knowledge extraction from multiple bootstrapped gene set enrichment analysis runs. A comparison of the two approaches in sample preparation for high-throughput sequencing shows that alternating decision trees represent the optimal knowledge representation method in comparison with classical decision trees.ConclusionsUsual reproducibility measurements are mostly based on statistical techniques that offer very limited biological insights into the studied gene expression data sets. This paper introduces the meta-learning-based gene set enrichment analysis that can be used to complement the analysis of gene-ranking stability estimation techniques such as percentage of overlapping genes or classic gene set enrichment analysis. It is useful and practical when reproducibility of gene ranking results or different gene selection techniques is observed. The proposed method reveals very accurate descriptive models that capture the co-enrichment of gene sets which are differently enriched in the compared data sets.

Headspace solid-phase microextraction followed by gas chromatography tandem mass spectrometry for the sensitive determination of benzotriazole UV stabilizers in water samples

A sensitive procedure for the determination of five ultraviolet (UV) absorbers, belonging to the benzotriazole class, in environmental water samples is proposed. Analytes were first extracted and concentrated from the matrix and then selectively determined by gas chromatography in combination with tandem mass spectrometry detection. The high lipophilic character of some of the investigated species resulted in a strong trend to remain sorbed on solid surfaces, even after addition of considerable percentages of methanol (up to 30%) to water. Thus, minimizing sample handling during the enrichment step is mandatory in order to obtain acceptable accuracy and precision. Solid-phase microextraction (SPME), as sample preparation approach, fulfilled the above requirement and provided acceptable figures of merit for the determination of target species in environmental water samples, including raw wastewater. Optimization of SPME conditions showed that the combination of headspace extraction, with a sample temperature of 100 degrees C and addition of 15 mg of NaCl per milliliter of sample rendered the best compromise in terms of extraction efficiency for all species. Considering a sampling time of 30 min with a poly(dimethylsiloxane)-divinylbenzene-coated SPME fiber, limits of quantification below 2 ng l(-1) and relative standard deviations between 5% and 12% were achieved. Three of the five species included in this research were determined in raw wastewater with a maximum concentration of 57 ng l(-1) for the Tinuvin 326 UV absorber.

Comparison of QuEChERS sample preparation methods for the analysis of pesticide residues in fruits and vegetables

This article describes the comparison of different versions of an easy, rapid and low-cost sample preparation approach for the determination of pesticide residues in fruits and vegetables by concurrent use of gas and liquid chromatography (GC and LC) coupled to mass spectrometry (MS) for detection. The sample preparation approach is known as QuEChERS, which stands for “quick, easy, cheap, effective, rugged and safe”. The three compared versions were based on the original unbuffered method, which was first published in 2003, and two interlaboratory validated versions: AOAC Official Method 2007.01, which uses acetate buffering, and European Committee for Standardization (CEN) Standard Method EN 15662, which calls for citrate buffering. LC–MS/MS and GC–MS analyses using each method were tested from 50 to 1000 ng/g in apple–blueberry sauce, peas and limes spiked with 32 representative pesticides. As expected, the results were excellent (overall average of 98% recoveries with 10% RSD) using all 3 versions, except the unbuffered method gave somewhat lower recoveries for the few pH-dependent pesticides. The different methods worked equally well for all matrices tested with equivalent amounts of matrix co-extractives measured, matrix effects on quantification and chemical noise from matrix in the chromatographic backgrounds. The acetate-buffered version gave higher and more consistent recoveries for pymetrozine than the other versions in all 3 matrices and for thiabendazole in limes. None of the versions consistently worked well for chlorothalonil, folpet or tolylfluanid in peas, but the acetate-buffered method gave better results for screening of those pesticides. Also, due to the recent shortage in acetonitrile (MeCN), ethyl acetate (EtOAc) was evaluated as a substitute solvent in the acetate-buffered QuEChERS version, but it generally led to less clean extracts and lower recoveries of pymetrozine, thiabendazole, acephate, methamidophos, omethoate and dimethoate. In summary, the acetate-buffered version of QuEChERS using MeCN exhibited advantages compared to the other tested methods in the study.

Use of the modified quick easy cheap effective rugged and safe sample preparation approach for the simultaneous analysis of type A- and B-trichothecenes in wheat flour

A suitable extraction and purification method for the simultaneous liquid chromatography–mass spectrometry (LC–MS) determination of five mycotoxins, three type A, diacetoxyscirpenol (DAS), T-2 toxin (T-2) and HT-2 toxin (HT-2), and two type B-trichothecenes, deoxynivalenol (DON) and nivalenol (NIV), has been optimised using a modified “Quick Easy Cheap Effective Rugged and Safe” (QuEChERS) method. Different solvents were studied in the extraction procedure to obtain better recoveries, which ranged from 86 to 108%, using a 85/15 (v/v) mixture of methanol/acetonitrile. The values obtained for recovery, repeatability and reproducibility of the optimized method are in agreement with Commission Directive 2005/26/EC for methods of analysis of Fusarium toxins. Finally, this optimized procedure was applied in wheat flour samples commercialized in Spain.

Determination strategies of phytohormones: recent advances

Phytohormones are key regulators in various physiological processes of plant growth and development. To better understand their regulatory actions, fast, sensitive and accurate methods for phytohormone analysis are critically required, and great efforts have been invested in this field. In this review, recent advances and developments in phytohormone determination strategies are presented, in terms of sample preparation and analytical approaches. Many types of analytical techniques are involved, including chromatography, mass spectrometry, immunoassay and others. Some challenges and prospects are also discussed.

Hydrophobic solvent induced phase transition extraction to extract drugs from plasma for high performance liquid chromatography–mass spectrometric analysis

Novel sample preparation approaches for HPLC bioanalysis based on the phenomenon that acetonitrile can be separated from water by adding salts or cooling at subzero temperatures have been reported. These two methods are superior to conventional liquid–liquid extraction since the separated acetonitrile phase can be directly injected to the RP–LC system. However, the salting-out method suffers from a potential problem that the remained salt in the acetonitrile phase may harm the MS detector, while the subzero-temperature method is troublesome to operate. Here, we have reported a similar phase separation phenomenon that the acetonitrile aqueous mixture can be separated by adding a hydrophobic solvent; and capitalising on this phase transition phenomenon, we have proposed an alternative approach, named solvent induced phase transition extraction (SIPTE), to extract drug from plasma for HPLC–MS analysis. The proposed SIPTE method is much simpler and avoids contaminating the MS detector. Three structurally diverse drugs were selected as test compounds to design the SIPTE method and to validate the efficiency of this method. The four goals of plasma sample pretreatment for HPLC–MS analysis, i.e. removal of proteins, removal of other low-molecular interferences, preconcentration of the analytes of interest, and matching the sample solvent with the HPLC–MS system, can be rapidly performed in a very simple step by using the SIPTE method.

Atom-Probe Tomography of Semiconductor Materials and Device Structures

AbstractThe development of laser-assisted atom-probe tomography (APT) analysis and new sample preparation approaches have led to significant advances in the characterization of semiconductor materials and device structures by APT. The high chemical sensitivity and three-dimensional spatial resolution of APT makes it uniquely capable of addressing challenges resulting from the continued shrinking of semiconductor device dimensions, the integration of new materials and interfaces, and the optimization of evolving fabrication processes. Particularly pressing concerns include the variability in device performance due to discrete impurity atom distributions, the phase and interface stability in contacts and gate dielectrics, and the validation of simulations of impurity diffusion. This overview of APT of semiconductors features research on metal-silicide contact formation and phase control, silicon field-effect transistors, and silicon and germanium nanowires. Work on silicide contacts to silicon is reviewed to demonstrate impurity characterization in small volumes and indicate how APT can facilitate defect mitigation and process optimization. Impurity contour analysis of a pFET semiconductor demonstrates the site-specificity that is achievable with current APTs and highlights complex device challenges that can be uniquely addressed. Finally, research on semiconducting nanowires and nanowire heterostructures demonstrates the potential for analysis of materials derived from bottom-up synthesis methods.

Development and validation of a multi-residue method for the determination of pesticides in honeybees using acetonitrile-based extraction and gas chromatography–tandem quadrupole mass spectrometry

An optimized analytical method employing gas chromatography–tandem quadrupole mass spectrometry (GC–MS/MS) has been developed for the simultaneous screening of roughly 150 pesticides in honeybees suspected of poisoning by pesticides during field spraying. In this work, a sample preparation approach based on acetonitrile extraction followed by dispersive solid-phase extraction (d-SPE) cleanup was implemented and validated for pesticides in honeybees for the first time. The procedure involved homogenization of a 2 g sample (23 insects on average) with acetonitrile–water mixture followed by salting out with citrate buffer, magnesium sulphate and sodium chloride. An amount of matrix constituents with limited solubility in acetonitrile was reduced in the extract by precipitation at low-temperature (freezing-out cleanup). Hereafter, d-SPE cleanup was carried out using primary secondary amine (PSA), octadecyl (C18) and graphitized carbon black (GCB). This combination of cleanup steps ensured efficient extract purification. Linearity of the calibration curves was studied using matrix-matched standards in the concentration range between 4 and 500 ng mL −1 (equivalent to 10 and 1250 ng g −1), and coefficients of determination ( R 2) were ≥0.99 for approximately 90% of the targeted compounds. The recovery data were obtained by spiking honeybees samples free of pesticides at three concentration levels of 10, 50, and 500 ng g −1 (approximately 0.9, 4.3, 43.5 ng per bee). At these spiking levels 47, 77 and 92% of the targeted compounds were recovered, respectively. Generally the recoveries were in the range between 70 and 120% with precision values, expressed as relative standard deviation (RSD) ≤ 20%. The expanded uncertainty was estimated following a “top down” empirical model as being 28% on average (coverage factor k = 2, confidence level 95%). Preliminary results from practical application to analysis of real samples are presented. A total of 25 samples of honeybees from suspected pesticides poisoning incidents were analyzed, in which 10 different pesticides were determined.

F-27 Water Analysis Using a Prototype Monochromatic Microfocus Grazing Incidence X-ray Fluorescence Device Comparing Picoliter and Nanoliter Deposition as Sample Preparation Approach

F-27 Water Analysis Using a Prototype Monochromatic Microfocus Grazing Incidence X-ray Fluorescence Device Comparing Picoliter and Nanoliter Deposition as Sample Preparation Approach - Volume 24 Issue 2

In vitro and in vivo investigation of metabolic fate of rifampicin using an optimized sample preparation approach and modern tools of liquid chromatography–mass spectrometry

Biotransformation products of rifampicin formed in vitro and in vivo were identified using modern tools of liquid chromatography–mass spectrometry (LC–MS). For the same, first the mass fragmentation pattern of rifampicin was delineated by conducting multiple-stage MS (MS n ), high resolution MS (HR-MS) and hydrogen/deuterium exchange MS (HDE-MS) experiments in an electro-spray ionization (ESI) mode. Then in vitro metabolism was investigated by incubating the drug with rat liver S9 fraction and microsomes at pH 7.4 in the presence of NADPH for 2 h at 37 °C. In vivo metabolites were generated by administering the drug to Sprague–Dawley (SD) rats ( n = 5) at a dose of 50 mg kg −1 and collecting blood, urine and feaces at different time points up to 72 h. The samples were processed using an optimized sample preparation approach, involving protein-precipitation using acetonitrile; followed by liquid-freeze separation at −20 °C; and solid-phase extraction of aqueous layer. HPLC conditions were optimized using known standards of rifampicin metabolites or degradants. The extracted and concentrated samples were then subjected to LC-DAD–MS/time-of-flight (LC-DAD–MS/TOF) analyses in both ESI positive and negative modes. The structures of metabolites were elucidated by comparing their accurate mass values and fragmentation patterns with those of rifampicin, use of in silico identification tools and relative retention time matching with the standards. The nature of the metabolites and their relative amounts varied. In total, 21 metabolites were identified, of which many structures were new, not reported till date.

Pressurized liquid extraction followed by gas chromatography with atomic emission detection for the determination of fenbutatin oxide in soil samples

A novel method for the determination of the miticide bis[tris(2-methyl-2-phenylpropyl)tin] oxide, also known as fenbutatin oxide (FBTO), in agricultural soils is presented. Pressurized liquid extraction (PLE) followed by analyte derivatization and extraction into isooctane was the used sample preparation approach. Selective determination was achieved by gas chromatography with atomic emission detection (GC-AED). Influence of different parameters on the performance of the extraction process is thoroughly discussed; moreover, some relevant aspects related to derivatization, determination and quantification steps are also presented. As regards PLE, the type of solvent and the temperature were the most relevant variables. Under optimized conditions, acetone, without any acidic modifier, was employed as extractant at 80°C. Cells were pressurized at 1500psi, and 2 static cycles of 1min each were applied. Acetone extracts (ca. 25mL) were concentrated to 1mL, derivatized with sodium tetraethyl borate (NaBEt4) and the FBTO derivative, resulting from cleavage of the Sn–O–Sn bond followed by ethylation of the hydroxyl fragments, extracted into isooctane and determined by GC-AED. Under final working conditions, the proposed method provided recoveries from 76 to 99% for spiked soil samples, a limit of quantification of 2ngg−1 and an acceptable precision. Analysis of samples from vineyards sprayed with FBTO, confirmed the persistence of the miticide in soil for more than 1 year after being applied.

Methods for the Analysis of Fluoroquinolones in Biological Fluids

Methods for the analysis of ten selected fluoroquinolone antibiotics in biological fluids are reviewed. Approaches for sample preparation, detection methods, limits of detection and quantitation, and recovery information are provided for both single analyte and multi-analyte fluoroquinolone methods.

Neuropeptidomics of the Supraoptic Rat Nucleus

The mammalian supraoptic nucleus (SON) is a neuroendocrine center in the brain regulating a variety of physiological functions. Within the SON, peptidergic magnocellular neurons that project to the neurohypophysis (posterior pituitary) are involved in controlling osmotic balance, lactation, and parturition, partly through secretion of signaling peptides such as oxytocin and vasopressin into the blood. An improved understanding of SON activity and function requires identification and characterization of the peptides used by the SON. Here, small-volume sample preparation approaches are optimized for neuropeptidomic studies of isolated SON samples ranging from entire nuclei down to single magnocellular neurons. Unlike most previous mammalian peptidome studies, tissues are not immediately heated or microwaved. SON samples are obtained from ex vivo brain slice preparations via tissue punch and the samples processed through sequential steps of peptide extraction. Analyses of the samples via liquid chromatography mass spectrometry and tandem mass spectrometry result in the identification of 85 peptides, including 20 unique peptides from known prohormones. As the sample size is further reduced, the depth of peptide coverage decreases; however, even from individually isolated magnocellular neuroendocrine cells, vasopressin and several other peptides are detected.

A high-throughput approach for cross-sectional transmission electron microscopy sample preparation of thin films

Cross-sectional transmission electron microscopy (XTEM) is a very useful technique to study the interfacial diffusion and reactions and the grain growth of thin films. However, the preparation of XTEM samples of thin films is tedious and challenging. Difficulties may include the delamination of films from the substrate, fracture of brittle substrates and differential milling rates of the substrate and the film. This paper describes an improved technique using a combination of tripod polishing and focused ion beam milling to prepare XTEM samples of thin films. The technique can be widely used for high-throughput production of samples having varying film and substrate properties. Two different geometries are introduced. The first one is suitable for XTEM sample preparation of most films at a high yield rate, but with a limited view area. The other geometry is able to give a larger view area and is more suitable for thicker films. The technique is illustrated by an example of the sample preparation of Fe/Pt multilayer films on SiO2/Si substrates.