Establishment of signal-recovery functions for calculation of recovery factor. Application to monitoring of contaminant residues in vegetables by chemiluminescence detection

A new strategy is proposed for verifying if recovery factor is constant and independent of the real analyte content of samples. A signal-recovery function has been developed on the basis of measurement of spiked test samples before and after a pre-treatment step and considering, as starting point, a recent IUPAC recommendation which distinguishes between two terms—recovery factor, R, and apparent recovery, R*. Apparent recovery includes recovery factor and a new recovery term proposed in a previous paper by the authors, named calibration recovery, R ^C. The signal-recovery function is obtained directly from the measured analytical signals instead of from the concentrations, simplifying the calculations. A linear signal-recovery curve indicates that the recovery factor is constant in the analyte concentration range studied experimentally and, in this way, a single recovery factor can be calculated. The usefulness of the proposed method has been shown by quantification of the pesticide carbaryl by two different flow-injection analysis methods with chemiluminescent detection based on the luminol and TCPO systems. Good results were obtained from both methods.     

Simplified batch equilibration for D/H determination of non‐exchangeable hydrogen in solid organic material

Hydrogen isotopic analysis of organic materials has been widely applied in studies of paleoclimate, animal migration, forensics, food and flavor authentication, and the origin and diagenesis of organic matter. Hydrogen bound to carbon (C‐H) generally retains isotopic information about the water present during organic matter synthesis and associated biosynthetic fractionations, but hydrogen bound to other elements (O, S, or N) can readily exchange with atmospheric water vapor and reflects recent exposure to water or vapor. These two pools must be separated to obtain meaningful information from isotope ratios of organic materials. Previously published analytical methods either replace exchangeable H chemically or control its isotopic composition, usually by equilibration with water or waters of known isotopic composition. In addition, the fraction of H that is exchangeable can vary among samples and is itself of scientific interest. Here we report an improved and automated double‐equilibration approach. Samples are loaded in a 50‐position autosampler carousel in an air‐tight aluminum equilibration chamber. Water vapor of known isotopic composition is pumped through the chamber at 115°C for at least 6 h. After flushing with dry N₂ and being cooled, the carousel is rapidly transferred from the equilibration chamber to a He‐purged autosampler attached to a pyrolysis elemental analyzer connected to an isotope ratio mass spectrometer. By equilibrating two aliquots of each sample with two isotopically distinct waters, it is possible to calculate both (1) the D/H ratio of non‐exchangeable H, and (2) the fraction of H that is exchangeable. Relative to previous double‐equilibration techniques, this approach offers significant reductions in sample size and labor by allowing simultaneous equilibration of several tens of samples. Copyright © 2009 John Wiley & Sons, Ltd.     

Ion chromatographic system with a novel switching suppression device

Ion chromatography (IC) has been a powerful tool for measuring ionic species in environmental samples such as tap, river and drain waters. Suppressor modules (membrane and disposable column types) have been used for reducing the background of a baseline. A new type of suppressor device, which has a suppressor resin and switching valve was developed. Fresh ionic resin is introduced into a groove for each analysis to perform the suppression of the working eluent. The eluent composition for obtaining higher sensitivity and better resolutions among ionic species and carbonate ion was also investigated. Although carbonate buffers are used for ion separation in general, the separation of carbonate ion from other ions was not achieved. A borate eluent resulted in good resolutions and higher sensitivity. A new column was also developed for obtaining higher column efficiency and resolution. The optimization of anion separation using a new IC system (IC-2001) that consists of a new suppressor device, an anion-exchange column (TSKgel SuperIC-Anion, 150x4.6 mm), an autosampler, a conductivity cell and a pump in a compact module is described.     

Automated at-column injection into narrow bore capillary GC columns

An injector designed for automatic direct liquid injection into narrow bore capillary GC columns has been constructed and evaluated. The tip of the syringe needle is aligned with, and positioned close to, the column entrance in a small, pressurized cavity: when the sample is dispensed it is immediately forced into the column by the action of the surrounding carrier gas. A standard autosampler equipped with a standard stainless steel syringe needle was utilized for at-column sample transfer into 100 μm i.d. columns. RSD values for n-alkanes were between 0.1 and 0.3% for relative area counts and approximately 1% for absolute area counts.     

A fast microwave-assisted,acid-vapor,steam-cleaning procedure for autosampler cups

A new and fast procedure is proposed for cleaning autosampler cups using acid-vapor steam-cleaning with a miniaturized assembly in a microwave-heated sealed Teflon vessel. A glass cactus-shaped holder was made to support six polyethylene autosampler cups (volume, 2.0 mL) inside a 100 mL microwave vessel. Regent-grade nitric acid was added to the vessels, and the system was heated in a microwave oven for 5 min at 300 W. Chromium was determined by graphite-furnace atomic-absorption spectrophotometry. The blank values were lower with cleaned cups compared to untreated cups (i.e., as received from supplier). The quantification limits, estimated from detection limits established with Milli-Q water, were 0.66 and 0.95 μg Cr L⁻¹ for cleaned and untreated auto-sampler cups, respectively.     

A system for the automated static headspace analysis of dissolved N2O in seawater

A new automated static headspace N₂O analysis technique has been developed that includes a CTC autosampler composed of agitator and headspace modules, a combined-valve switching system and ECD Gas Chromatography. This new CTC autosampling technique is more efficient for sample analysis than similar systems and takes approximately 10 mins to analyze each sample without pre-equilibration. The accuracy and precision of this method are approximately 2% each. Laboratory and field results demonstrate that this technique is suitable for seawater sample analysis.     

Recent developments in solid-phase microextraction

The main objective of this review is to describe the recent developments in solid-phase microextraction technology in food, environmental and bioanalytical chemistry applications. We briefly introduce the historical perspective on the very early work associated with the development of theoretical principles of SPME, but particular emphasis is placed on the more recent developments in the area of automation, high-throughput analysis, SPME method optimization approaches and construction of new SPME devices and their applications. The area of SPME automation for both GC and LC applications is particularly addressed in this review, as the most recent developments in this field have allowed the use of this technology for high-throughput applications. The development of new autosamplers with SPME compatibility and new-generation metal fibre assemblies has enhanced sample throughput for SPME-GC applications, the latter being attributed to the possibility of using the same fibre for several hundred extraction/injection cycles. For LC applications, high-throughput analysis (>1,000 samples per day) can be achieved for the first time with a multi-SPME autosampler which uses multi-well plate technology and allows SPME sample preparation of up to 96 samples in parallel. The development and evolution of new SPME devices such as needle trap, thin-film microextraction and cold-fibre headspace SPME have offered significant improvements in performance characteristics compared with the conventional fibre-SPME arrangement. Figure Photo of a high-throughput multi-fibre SPME PAS autosampler     

A simple and sensitive assay for eflornithine quantification in rat brain using pre‐column derivatization and UPLC‐MS/MS detection

Eflornithine (α‐difluoromethylornithine) has been used to treat second‐stage (or meningoencephalitic‐stage) human African trypanosomiasis and currently is under clinical development for cancer prevention. In this study, a new ultraperformance liquid chromatography–tandem mass spectrometry (UPLC‐MS/MS)‐based assay was developed and validated for the quantification of eflornithine in rat brain. To improve chromatographic retention and MS detection, eflornithine was derivatized with 6‐aminoquinolyl‐N‐hydroxysuccinimidyl carbamate for 5 min at room temperature prior to injection. Derivatized eflornithine was separated on a reverse‐phase C₁₈ UPLC column with a 6‐min gradient; elution occurred at approximately 1.5 min. Prior to derivatization, eflornithine was reproducibly extracted from rat brain homogenate by methanol protein precipitation (~70% recovery). Derivatized eflornithine was stable in the autosampler (6 °C) for at least 24 h. This new assay had acceptable intra‐ and interday accuracy and precision over a wide dynamic range (5000‐fold) and excellent sensitivity with a lower limit of quantification of 0.1 µm (18 ng/mL) using only 10 μL of rat brain homogenate. The validated eflornithine assay was applied successfully to determine eflornithine distribution in different regions of rat brain in an in situ rat brain perfusion study. Copyright © 2014 John Wiley & Sons, Ltd.     

Predicting eutectic behavior of drugs and excipients by unique calculations

A eutectic is formed from a mixture of two or more solids and has a melting point lower than that of each of its constituents. It is generally represented by a phase diagram where the liquid and solid phases impact upon each other with a value known as the eutectic point. In pharmaceuticals, poor water solubility is a major obstacle for releasing new dosage forms into the market. Eutectic formation overcomes these problems. Preparation of a phase diagram by Differential Scanning Calorimetry can determine eutectic properties, but it is tedious. A modified Van’t Hoff (VH) equation was used in this study. Devalina Law developed a dimensionless index for the VH equation. The difference in melting points of an excipient polymer and drug are divided by the slope of the VH equation. In previous studies, five excipient–drug compositions were evaluated. The final index relationship was in good agreement except for the salt, quinine sulfate. In order to test the validity of the VH index, further studies of PEG with acetylsalicylic acid, acetaminophen, diflunisal, dimenhydrinate, ketoconazole, and mefenamic acid were performed.     

Optimization of the cold split-splitless injector in the solvent-split mode

In this study, the cold split-splitless injector recently introduced by Carlo Erba was optimized for the trace analysis of polycyclic aromatic hydrocarbons. Injections of hundreds of microliters of hexane solutions were performed in the solvent split mode using an autosampler programmed for large volume injection. Injection parameters such as type of insert, initial oven temperature, split time, and flow are shown to play important roles in defining the conditions for obtaining quantitative results. The loss of early eluting compounds can be minimized by achieving a solvent trapping effect, making use of n-octane as co-solvent in n-hexane.     

Coupled in situ electrodeposition-electrothermal atomic absorption spectrometry: a new approach in quantitative matrix free analysis

A new approach for in situ matrix elimination in electrothermal atomic absorption spectroscopy (ETAAS) is described. In an initial electrodeposition step (possible by use of a Pt/Ir delivery tube on the autosampler) the furnace is coated with about 0.25 μg Pd. Quantitative deposition of metallic analytes onto this renewable substrate is achieved from 25–40 μl samples by electrolysis for 60 s at 3.5–5.0 V (35–45 mA). Reprogramming of the autosampler to remove spent electrolyte after the electrolyses and to provide a rinse cycle facilitates removal of > 99.5% of a 0.5 M NaCl matrix prior to atomization. It is proposed that the analyte is bound onto the metallic modifier, rather than encapsulated within it. Binding of the analyte with Pd significantly increases the appearance temperature for Cd and Pb. The ashing loss for these analytes deposited onto Pd from a Cl⁻ matrix is observed above 900°C and 1300°C, respectively. This stabilization facilitates separation of the residual NaCl matrix before atomization. It has been established for Cd that sensitivity of the determination remains constant for matrices as diverse as 1% HNO₃, 0.5 M NaCl and sea water.     

Quantum chemical structure–activity relationships on β-carbolines as natural monoamine oxidase inhibitors

The electron density and the molecular electrostatic potential of the β-carbolines are studied using ab initio STO -3G wave functions. The analysis was done from the point of view of a previous model built with monoamine oxidase substrates and irreversible inhibitors. The results confirm the usefulness of the model and make it possible to propose new precision to the molecular electrostatic potential patterns needed to have monoamine oxidase inhibitory activity.     

Upward extrapolation of saturated liquid densities

The design of new energy conversion processes requires equations of state for the working fluids. For their construction saturated liquid densities are needed which are not available for some potential working fluids at higher temperatures. Hence, we investigate how Rackett-type equations behave in extrapolations from saturated liquid densities in the temperature range 0.5 ≤ T/T_c ≤ 0.75 up to the critical temperature T_c. Extrapolation methods with different inputs from the critical point are used: (a) no critical point data, (b) critical temperature, (c) critical temperature and pressure, and (d) critical temperature and compression factor. It is found that upward extrapolations of the saturated liquid densities without using critical point data can be done with some care and that the additional use of the critical temperature improves the quality of the predictions substantially.     

Automated in-tube solid-phase microextraction-high-performance liquid chromatography for carbamate pesticide analysis

In-tube solid-phase microextraction (SPME) is an automated version of SPME that can be easily coupled to a conventional HPLC autosampler for on-line sample preparation, separation and quantitation. It has been termed "in-tube" SPME because the extraction phase is coated inside a section of fused-silica tubing rather than coated on the surface of a fused-silica rod as in the conventional syringe-like SPME device. The new in-tube SPME technique has been demonstrated as a very efficient extraction method for the analysis of polar and thermally labile analytes. The in-tube SPME-HPLC method used with the FAMOS autosampler from LC Packings was developed for detecting polar carbamate pesticides in clean water samples. The main parameters relating to the extraction and desorption processes of in-tube SPME (selection of coatings, aspirate/dispense steps, selection of the desorption solvents, and the efficiency of desorption solvent, etc.) were investigated. The method was evaluated according to the reproducibility, linear range and limit of detection. This method is simple, effective, reproducible and sensitive. The relative standard deviation for all the carbamates investigated was between 1.7 and 5.3%. The method showed good linearity between 5 and 10000 microg/l with correlation coefficients between 0.9824 and 0.9995. For the carbamates studied, the limits of detection observed are lower than or similar to that of US Environmental Protection Agency or National Pesticide Survey methods. Detection of carbaryl present in clean water samples at 1 microg/l is possible.     

“Groove-injection” as a novel method for sub-microliter (nanoliter) injections in liquid chromatography using a conventional multi-port valve

In liquid chromatography with “low-dispersion methods”, there is an increasing need to reproducibly inject nanoliter sample volumes. Low-dispersion methods produce very narrow peaks because of short column length, narrow column bore, small particle packing, low particle surface area, open tubular configuration, or combinations of these parameters. This paper describes a new injector method, the “groove-injector” which involves simple plumbing changes for use of a conventional multi-port valve (8-ports or more) to inject sample volumes approximating a single groove in such a valve (e.g 30 nanoliters with aceptable reproducibility, ca. 8% RSD). In addition, by changing a resistor, volumes between 30, and over 2,000 nanoliters (nearly two orders of magnitude) can be injected with reproducibilities generally below 2% RSD. Different samples can be injected by using an autosampler. Compared to commonly used 4-port valves for nanoliter injections, multi-port valves have a number of advantages. Multiport valves generally are more commonly available and they are a better financial investment because of their versatility for column switching, sample enrichment, or variable volume injections. Previously, submicroliter (nanoliter) injections have not been possible with multi-port valves in as direct and simple a manner as described here.     

Optimizing an ultrafast generic high-performance liquid chromatography/tandem mass spectrometry method for faster discovery pharmacokinetic sample throughput

For higher throughput screening, where the number of new chemical entities (NCEs) to test is rapidly increasing, fast sample turnaround time is essential. In order to increase efficiency a generic high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) method, with a cycle time of 85 s (42 injections/h), was created. This was accomplished through the use of a 1-min ballistic gradient and the optimization of the autosampler. The gradient was optimized by varying the organic mobile phase concentration and examining its ballistic characteristics with respect to matrix ion suppression and compound retention time. The autosampler time could be reduced by optimizing several parameters and by determining the source of most of the carryover in order to reduce the number of syringe and injector washes. Finally, the reliability of the new generic method is demonstrated by comparison of sample data with a standard 2-min linear gradient method that showed that the data sets were well correlated. For plasma AUC (ng.h/mL) of 28 NCEs, the regression line had a slope of 0.92 and the R2 was 0.929. The described method was found to be useful for both rat plasma and tissue samples.     

Efficient Total Synthesis of (S)‐14‐Azacamptothecin

An efficient total synthesis of (S)‐14‐azacamptothecin has been accomplished in 10 steps and 56 % overall yield from 5H‐pyrano[4,3‐d]pyrimidine 8 . A mild Hendrickson reagent‐triggered intramolecular cascade cyclization, a highly enantioselective dihydroxylation, and an efficient palladium‐catalyzed transformation of an O‐allyl into N‐allyl group are the key steps in the synthesis. This work provides a much higher overall yield than the previous achievement and shows sound flexibility for the further applications that will lead to new bioactive analogues.     

Quantitative determination of methylphenidate in plasma by gas chromatography negative ion chemical ionisation mass spectrometry using <Emphasis Type="Italic">o</Emphasis>-(pentafluorobenzyloxycarbonyl)-benzoyl derivatives

The use of a novel electrophoric derivatisation reagent, o-(pentafluorobenzyloxycarbonyl)-benzoyl chloride, for the quantitative determination of methylphenidate in plasma is described. The drug can be quantitatively measured down to 72 pg/mL plasma using only 250 μL of sample due to the extraordinary sensitivity of the derivatives under negative ion chemical ionisation mass spectrometry. Plasma samples were made alkaline with carbonate buffer and treated with extraction solvent n-hexane and reagent solution for 30 min, which, after concentration, was measured by GC-NICI-MS. The method is rapid as extraction and derivatisation occur in one single step. A stable isotope-labelled internal standard was used and its synthesis described. Full validation data are given to demonstrate the usefulness of the assay, including specificity, linearity, accuracy and precision, long-term stability, short-term stability, freeze–thaw stability, stock solution stability, autosampler stability, aliquot analysis, robustness, matrix effect, and prospective analytical batch size accuracy. The method has been successfully applied to pharmacokinetic profiling of the drug after oral application.     

Generative topographic mapping of binding pocket of β2 receptor and three‐way partial least squares modeling of inhibitory activities

The visualization and characterization of protein pockets is the starting point for many structure‐based drug design projects. The size and shape of protein pockets dictate 3D geometry of ligands that can strongly inhibit the following biological events. Thus, a minimal requirement for inhibition is that a molecule sterically binds the active site with some allowance for induced fit. Methods for direct display of active sites in a protein have become prevalent in recent years. In this study, a new mapping method, generative topographic mapping, is investigated to describe the 3D surface of protein pocket. The β2 receptor protein is used as a benchmark. By mapping the molecular surface points and assigning the associated molecular electrostatic potential (MEP) values, the original 3D structure of the active site is well reproduced by the 2D latent map in generative topographic mapping. The distributions of MEP values of two 2D latent maps derived from the inhibitor and the β2 receptor protein are well complemented. Using three‐way partial least squares modeling, a predictive model linking the inhibitory activity and their MEP values can be constructed, which was not feasible in the previous spherical self‐organizing map studies. The resulting regression coefficient matrix of the three‐way partial least squares model has many insights for understanding the structural requirements for β2 inhibitory activity. Copyright © 2014 John Wiley & Sons, Ltd.     

Automated high throughput analysis of antiretroviral drugs in dried blood spots

For therapeutic drug monitoring in remote settings, dried blood spots (DBS) are particularly advantageous, as blood sample collection and handling is uncomplicated. The aim of this study was to develop and validate an automated extraction method for the analysis of nevirapine, efavirenz and lopinavir in DBS samples. Automated extraction was performed with methanol : water (70 : 30 v /v ), using a DBS‐MS 500 autosampler coupled to a liquid chromatography tandem mass spectrometry system. The autosampler used digital images of each DBS to position the extraction head, sprayed 10 μl of internal standard onto each DBS and extracted a 4‐mm disc (Ø) from the centre of each spot by unilateral flow using 25‐μl extraction solvent. The analytes were baseline separated on a pentafluorophenyl column and analysed by using electrospray ionization with multiple reaction monitoring in positive polarity mode for nevirapine and lopinavir and in negative mode for efavirenz. The method was linear between 10 and 10 000 ng/ml for all analytes. Automated sample extraction resulted in consistent recoveries (nevirapine: 70 ± 6%, efavirenz: 63 ± 11% and lopinavir: 60 ± 10%) and matrix effects between different donors and concentration levels. Intra‐day and inter‐day accuracy and precision deviations were ≤15%. Manual and automated extractions of DBS samples collected within the framework of an adherence assessment study in rural Tanzania showed good agreements with deviations of less than 10%. Our study highlights that therapeutic drug monitoring samples obtained in the resource‐constrained setting of rural Africa can be reliably determined by automated extraction of DBS. Overall, automatization improved method sensitivity and facilitates analysis of large sample numbers. Copyright © 2017 John Wiley & Sons, Ltd.