Automated high-capacity sorption probe for extraction of organic compounds in aqueous samples followed by gas chromatographic analysis

An automated high-capacity sorption device for GC analysis of ultra trace components has been developed. The scope of the presented technique was to combine the simplicity of solid-phase microextraction (SPME) with the high extraction efficiency of the stir bar sorptive extraction technology. Sorptive extractions of water samples were performed using polydimethylsiloxane (PDMS) rubber tubing (120 microl) mounted onto a glass rod. The sampling procedure was carried out by a robotic autoinjector. Since the setup is fully automated, unattended and precise time-controlled extraction of samples is possible and makes quantitation with non-equilibrium extractions feasible. The sorption probes are easy to exchange, which facilitates off-line/in-field sampling. The system was evaluated with a test mixture of 44 environmentally hazardous compounds. Detection limits were found to be in the sub-ppt region. The performance of the system was demonstrated with the analysis of polycyclic aromatic hydrocarbons in urban snow.     

Multiple solid-phase microextraction of drugs from human urine

Summary The applicability of multiple solid-phase microextraction to the analysis of biological samples has been shown by extraction of a variety of compounds from human urine. Multiple solid-phase microextraction, in which extraction and desorption are repeated and analytes are collected at the head of the separation system before starting the analysis, has been combined with gas chromatography. Amphetamine, lidocaine, procaine, and mepivacaine were extracted from buffered urine by direct immersion of a 100-μm polydimethylsiloxane-coated fiber, to demonstrate that multiple SPME can be used for analytes with different extraction behavior. Multiple solid-phase microextraction was optimized for high extraction yield or short extraction time. For example, the total sample-handling time (extraction plus desorption) for the extraction of mepivacaine from urine can be reduced from approximately 60 min (one extraction) to 33 min (three extractions) without reducing extraction yield. In addition, the extraction yield for mepivacaine can be increased from 14.6% (one extraction) to 27.0% (five extractions) within the same total sample handling time of approximately 60 min. A good match between theoretical and experimental values was obtained. Chromatograms are shown to illustrate the usefulness of the procedure.     

Monolithic methacrylate packed 96-tips for high throughput bioanalysis

In the pharmaceutical industry the growing number of samples to be analyzed requires high throughput and fully automated analytical techniques. Commonly used sample-preparation methods are solid-phase extraction (SPE), liquid–liquid extraction (LLE) and protein precipitation. In this paper we will discus a new sample-preparation technique based on SPE for high throughput drug extraction developed and used by our group. This new sample-preparation method is based on monolithic methacrylate polymer as packing sorbent for 96-tip robotic device. Using this device a 96-well plate could be handled in 2–4 min. The key aspect of the monolithic phase is that monolithic material can offer both good binding capacity and low back-pressure properties compared to e.g. silica phases. The present paper presents the successful application of monolithic 96-tips and LC–MS/MS by the sample preparation of busulphan, rescovitine, metoprolol, pindolol and local anaesthetics from human plasma samples and cyklophosphamid from mice blood samples.     

New cold-fiber headspace solid-phase microextraction device for quantitative extraction of polycyclic aromatic hydrocarbons in sediment

A new automated headspace solid-phase microextraction (HS-SPME) sampling device was developed, with the capability of heating the sample matrix and simultaneously cooling the fiber coating. The device was evaluated for the quantitative extraction of polycyclic aromatic hydrocarbons (PAHs) from solid matrices. The proposed device improves the efficiency of the release of analytes from the matrix, facilitates the mass transfer into the headspace and significantly increases the partition coefficients of the analytes, by creating a temperature gap between the cold-fiber (CF) coating and the hot headspace. The reliability and applicability of previously reported cold-fiber devices are significantly enhanced by this improvement. In addition, it can be easily adopted for full automation of extraction, enrichment and introduction of different samples using commercially available autosampling devices. Sand samples spiked with PAHs were used as solid matrices and the effect of different experimental parameters were studied, including the extraction temperature, extraction time, moisture content, and the effect of sonication and modifier under optimal experimental conditions, linear calibration curves were obtained in the range of 0.0009-1000 ng/g, with regression coefficients higher than 0.99 and detection limits that ranged from 0.3 to 3 pg/g. Reproducible, precise and high throughput extraction, monitoring and quantification of PAHs were achieved with the automated cold-fiber headspace solid-phase microextraction (CF-HS-SPME) device coupled to GC-flame ionization detection. Determination of PAHs in certified reference sediments using the proposed approach exhibited acceptable agreement with the standard values.     

Before the injection--modern methods of sample preparation for separation techniques

The importance of sample preparation methods as the first stage in an analytical procedure is emphasised and examined. Examples are given of the extraction and concentration of analytes from solid, liquid and gas phase matrices, including solvent phase extractions, such as supercritical fluids and superheated water extraction, solid-phase extraction and solid-phase microextraction, headspace analysis and vapour trapping. The potential role of selective extraction methods, including molecular imprinted phases and affinity columns, are considered. For problem samples alternative approaches, such as derivatisation are discussed, and potential new approaches minimising sample preparation are noted.     

SynCar: an approach to automated synthesis

The automation of all aspects of manual solution-phase synthesis into one integrated, efficient, and reliable system could be regarded as something of an unmet challenge in organic chemistry. The requirements for modern solution-phase libraries in mainstream drug discovery is typically 50-250 high-purity compounds on a 10-100-mg scale, whether for target class libraries or lead optimization, and short cycle time in combination with high capacity is critical. To achieve these goals, in a codevelopment between Aventis and Accelab GmbH, Kusterdingen, Germany, we designed a completely novel system of independent workstations connected by a shuttle transfer system produced by Montech, Derendingen, Switzerland. Seven modular workstations process four reactions on each shuttle in parallel, with the ability to perform synthesis (temperature control and liquid reagent handling), filtration, liquid-liquid extraction, evaporation, weighing, solid-phase extraction, and HPLC/MS analysis. The modular design enables the continuous loading of shuttles at any time, and each shuttle can have its own workflow. The design also allows easy expansion for future needs. The result is a combination of high flexibility and high throughput.     

Automated sample preparation-fractionation for the measurement of dioxins and related compounds in biological matrices: a review

This article reviews some of the recent developments in the extraction and clean-up areas of biological samples dedicated to dioxin and related compound analysis. A brief introduction on the major dioxin contamination events, which have occurred in the food chain, is given to illustrate the need of fast high throughput methods in case of crises. The emphasis of this paper is the method development based upon reliable instrumental extraction techniques for rapid sample processing and automation such as; supercritical fluid extraction (SFE), microwave-assisted extraction (MAE), pressurized liquid extraction (PLE) and, solid-phase extraction (SPE). The PLE and SPE are also discussed in conjunction with the use of a multi-column automated clean-up system that can accommodate up to 5 g of extracted lipids. The fractionation in sub-groups of analytes during the clean-up process allows the isolation of various types of toxicants from a single sample and illustrates the versatility of the system. An integrated extraction and clean-up instrument is finally presented in terms of feasibility and attainable sample turnover for the parallel processing of liquid and solid biological samples.     

Analysis of free and bound phenolics in wine and grapes by GC–MS after automated SPE

The results of validation of a method for the analysis of free and bound phenolics in wine and grapes are presented. Wine and grape extracts are fractionated by automated solid-phase extraction on Bond Elut PPL cartridges to give free and bound phenolic fractions. Bound fractions are subjected to acid hydrolysis, and the phenolics released are recovered by solid-phase extraction on Bond Elut PPL cartridges. The fractions are further purified by automated solid-phase extraction on Bond Elut silica cartridges. After derivatisation to form trimethylsilyl ethers, the phenolics are determined by gas chromatography–mass spectrometry with selected ion monitoring. The method is suitable for robust, high-throughput monitoring of the concentrations of phenolics that can affect the palatability of wine.     

Task-specific microextractions using ionic liquids

Ionic liquids (ILs) have been the focus of many scientific investigations including the field of analytical microextractions. ILs have many advantages over traditional organic solvents making them excellent candidates as extraction media for a variety of microextraction techniques. Many physical properties of ILs can be varied, and the structural design and make-up can be tuned to impart desired functionality for enhancement of analyte extraction selectivity, efficiency, and sensitivity. This paper provides a brief overview of ionic liquids and highlights trends in three important sample-preparation techniques, namely, single drop microextraction, solid-phase microextraction, and dispersive liquid–liquid microextraction in terms of performing task-specific extractions using these highly versatile solvents.     

Solid-phase synthesis of five-dimensional libraries via a tandem Petasis-Ugi multi-component condensation reaction

Five-dimensional libraries of dipeptide amides are readily prepared using a solid-phase tandem Petasis-Ugi multi-component condensation protocol on either a RINK amine or Universal RINK isonitrile resin. The method is practical and can be automated to prepare a large number of compounds useful for high throughput biological screening.     

A gas chromatographic assay for quantitative analysis of volatiles in solid materials by discontinuous gas extraction

Summary A method has been developed for the quantitative analysis of volatile compounds in solid samples. The method is based on a stepwise gas extraction of the volatiles with subsequent analysis of the extracted material and is termed discontinuous gas extraction. Any quantitative analysis requires an exhaustive extraction, which, however, is often too time-consuming for routine analysis. It is shown how the total amount of each volatile compound can be calculated from only a few extractions. Such a calculation is possible because for analytical purposes it is the information of the extraction process and not the extracted material that is needed. This method is useful for samples which are insoluble, such as certain polymers or residual solvents in printed foils, and which cannot be analyzed quantitatively by headspace gas chromatography, since no calibration solution can be prepared. It is further shown how discontinuous gas extraction can also be used to calibrate headspace analysis. Thus, both methods combine well together in that discontinuous gas extraction provides the accuracy while the headspace analysis gives convenience and speed of sample throughput, particularly if carried out with an automated headspace analyzer.     

Automated and parallel microfluidic DNA extraction with integrated pneumatic microvalves/pumps and reusable open-channel columns

A novel microfluidic DNA extraction protocol based on integrated diaphragm microvalves/pumps and silica-deposited open-channel columns was developed specifically for automated and parallel DNA solid-phase extraction (SPE). The method uses microfluidic chips with a sandwiched structure containing three layers, which are the upper fluidic layer with surface-deposited silica on glass open channels as the extraction phase, the lower actuation layer with valve actuation channels on a glass wafer, and the middle poly(dimethylsiloxane) (PDMS) membrane for reversible bonding of the two glass substrates. These two glass substrates can be reused after thoroughly cleaning and the PDMS membrane can be replaced conveniently, which could effectively decrease the time and cost of chip manufacturing. The normally closed microvalves/pumps were used to automatically control all processes of the on-chip DNA SPE without cross-contamination and leakage, enabling the processing of multiple samples in parallel without changing the microvalve control module. Using the microchip device with integrated microvalves/pumps, automated, programmable, and simultaneous λ-DNA extractions from different samples could be attained, even from complex solutions such as human blood, and the silica-deposited open-channel columns could be reused stably and reliably. Results have demonstrated that most of the eluted λ-DNA was recovered in the second 2 µL of elution buffer with high-purity suitable for successful polymerase chain reaction amplification, making it possible for further integration into microfluidic devices for fully functional and high-throughput genetic analysis.     

Comparison of two extraction methods independently developed on two conceptually different automated supercritical fluid extraction systems for the determination of polychlorinated biphenyls in sediments

Two extraction methods that independently have been developed on conceptually different automated supercritical fluid extraction systems, ISCO SFX 3560 (syringe pump and liquid trapping) and Hewlett-Packard 7680T SFE (reciprocating pump and solid-phase trapping), were compared for the extraction of polychlorinated biphenyls from two Swedish sediments. The results demonstrated that the high-temperature ISCO method in some cases yields a more exhaustive extraction, but also less clean extracts due to co-extraction of unwanted matrix components which are all present in the trapping solvent. The medium-temperature Hewlett-Packard method may sometimes cause problems with quantitative recoveries, but on the other hand it yields very clean extracts due to the extra selectivity resulting from collection on a solid-phase trap.     

Chromatographic determination of vitamins in foods.

Chromatographic methods for the determination of water- and fat-soluble vitamins in foods are reviewed. For each vitamin, sample preparation, detection problems and chromatographic conditions are presented and discussed. High-performance liquid chromatography (HPLC) is becoming a standard method in vitamin assay, especially for routine work. HPLC systems can be automated using in-line solid-phase extraction and column switchings, resulting in very sensitive methods, even when simple UV detection is employed.     

Preparation and evaluation of a molecularly imprinted sol-gel material for on-line solid-phase extraction coupled with high performance liquid chromatography for the determination of trace pentachlorophenol in water samples

A highly selective imprinted amino-functionalized silica gel sorbent was prepared by combining a surface molecular imprinting technique with a sol-gel process for on-line solid-phase extraction-HPLC determination of trace pentachlorophenol (PCP) in water samples. The PCP-imprinted amino-functionalized silica sorbent was characterized by FT-IR, SEM, nitrogen adsorption and the static adsorption experiments. The imprinted functionalized silica gel sorbent exhibited high selectivity and offered a fast kinetics for the adsorption and desorption of PCP. The prepared sorbent was shown to be promising for on-line solid-phase extraction for HPLC determination of trace levels of PCP in environmental samples. With a sample loading flow rate of 5 ml min(-1) for 2 min, an enhancement factor of 670 and a detection limit (S/N = 3) of 6 ng l(-1) were achieved at a sample throughput of five samples h(-1). The precision (RSD) for nine replicate on-line sorbent extractions of 10 microgl(-1) PCP was 3.8%. The sorbent also offered good linearity (r = 0.9997) for on-line solid-phase extraction of trace levels of PCP. The method was applied to the determination of PCP in local lake water, river water and wastewater samples.     

Application of polyaniline–multiwalled carbon nanotubes composite fiber for determination of benzaldehyde in injectable pharmaceutical formulations by solid-phase microextraction GC–FID using experimental design

A polyaniline–multiwalled carbon nanotubes composite fiber was electrodeposited onto a platinum wire using cyclic voltammetry. This fiber was used for headspace solid-phase microextraction combined with gas chromatography and flame ionization detector of trace levels of benzaldehyde in some injectable pharmaceutical formulations. Three solid-phase microextraction parameters including temperature, time and stirring rate were investigated simultaneously using a three-level-three-factor Box–Behnken as experimental design on the extraction capability. The as-made fiber has a lifetime of over 300 extractions without any obvious decline in extraction efficiency. At the optimum conditions (extraction temperature 60°C, extraction time 15 min, stirring rate 700 rpm), the method displays excellent linearity over the concentration range of 25–1000 ng/mL of benzaldehyde with RSD values ranging from 1.0 to 6.8%. The limits of quantitation and detection were 25 and 10 ng/mL, respectively.     

Monolith as a new sample preparation material: recent devices and applications

Monolith was first used as a material for chromatographic separation two decades ago and solid-phase extraction over 10 years, and since then, separation science has undergone a dramatic change owing to advancements in analytical technology. Recently, monolith has been modified to suit various devices for the extraction and enrichment of analytes in any matrices of environmental, food, and biological analyses. This approach has contributed to miniaturization and automation for sample preparation, and it can reduce the time and cost requirements of sample preparation. Recently, numerous applications have been demonstrated for online and inline preconcentration coupled with monolith, and many kinds of devices have been designed and developed for offline devices. In this review, these applications and devices are listed and discussed in reference to other fields.     

Some applications of supercritical fluid extraction

Supercritical fluid extraction (SFE) exploits the solvation power of fluids at temperatures and pressures close to their critical point. Use of SFE with supercritical CO₂ is reported for the extraction of caffeine and quinine from various plant materials and of morphine from serum. Results are compared with those obtained by extractions with subcritical methanol and tetrahydrofuran, normal organic Soxhlet extractions and solid-phase extraction.