Leonard M. Sidisky

Trigonal tricationic ionic liquids: a generation of gas chromatographic stationary phases.

Trigonal tricationic ionic liquids (ILs) are a new class of ILs that appear to be unique when used as gas chromatographic stationary phases. They consist of four core structures; (1) A = mesitylene core, (2) B = benzene core, (3) C = triethylamine core, and (4) D = tri(2-hexanamido)ethylamine core; to which three identical imidazolium or phosphonium cationic moieties were attached. These were coated on fused silica capillaries, and their gas chromatographic properties were evaluated. They were characterized using a linear solvation parameter model and a number of test mixtures. On the basis of the literature, it is known that both monocationic and dicationic ILs possess almost identical polarities, solvation characteristics, and chromatographic selectivities. However, some of the trigonal tricationic ILs were quite different. The different solvation parameters and higher apparent polarities appear to generate from the more rigid trigonal geometry of these ILs, as well as their ability to retain the positive charges in relatively close proximity to one another in some cases. Their unique selectivities, retention behaviors, and separation efficiencies were demonstrated using the Grob mixture, a flavor and fragrance test mixture, alcohols/alkanes test, and FAME isomer separations. Two ILs C1 (methylimidazolium substitution) and C4 (2-hydroxyethylimidazolium substitution) had higher apparent polarities than any know IL (mono, di, and tricationic ILs) or commercial stationary phases. The tri(2-hexanamido)ethylamine core IL series proved to be very interesting in that it not only showed the highest separation efficiency for all test mixtures, but it also is the first IL stationary phase (containing NTf(2)(-) anions) that eliminates peak tailing for alcohols and other H-bonding analytes. The thermal stabilities were investigated using three methods: thermogravimetric analysis (TGA) method, temperature programmed gas chromatographic method (TPGC), and isothermal gas chromatographic method. The D core series had a high working temperature range, exceptional selectivities, and higher separation efficiencies than comparable polarity commercial columns. It appears that this specific type of multifunctional ILs may have the most promising future as a new generation of gas chromatographic stationary phases.

In Vivo Solid-Phase Microextraction: Capturing the Elusive Portion of Metabolome†

The main objective of metabolomics is the analysis of all lowmolecular-weight compounds present in a particular living system. Metabolomics data is complementary to proteomics, genomics, and transcriptomics data and provides a better understanding of dynamic processes occurring in living systems. The processes of sampling and sample preparation can significantly affect the composition of the measured metabolome, so the analytical results may not adequately reflect the true metabolome composition at the time of sampling. This is due primarily to poor efficiency (or even complete omission) of metabolism quenching step and multistep handling procedures, which contribute to inadvertent metabolite loss and/or degradation. Herein we introduce in vivo solid-phase microextraction (SPME) as a new sample preparation method for global metabolomics studies of living systems using liquid chromatography–mass spectrometry (LC-MS). SPME is a nonexhaustive sample preparation procedure in which the amount of analyte extracted is governed by the distribution coefficient of the analyte between the SPME coating and sample matrix if the equilibrium is reached or the rate of mass transfer if a short sampling time is used. In vivo SPME allows accurate extraction of the metabolome directly in the tissue or blood of freely moving animals without the need to withdraw a representative biological sample for analysis, under conditions of negligible depletion where the amount of analyte extracted by SPME is independent of the sample volume. The blood-draw-free nature of the sampling method facilitates multiple sampling of the same living system and can capture unstable or short-lived metabolites. Large biomolecules are not extracted by the specially selected biocompatible SPME coating, so the need for a metabolism quenching step is eliminated. The amount of metabolites extracted is proportional to the biologically active unbound concentration. For metabolomics studies, in vivo SPME provides the simplest and most rapid sample preparation tool available to date to study living systems in a format directly compatible with LC-MS detection. Although SPME was successfully applied to metabolomics studies using GCMS primarily in headspace mode, its capability to provide instantaneous metabolism quenching directly during the sampling process to capture true metabolome of blood or tissue has not been previously evaluated. First, we developed a successful in vivo SPME workflow for direct sampling of metabolome, and applied it to mice as a model system (Figure 1). In this approach, a coated SPME

Bonded ionic liquid polymeric material for solid-phase microextraction GC analysis

AbstractFour new ionic liquids (IL) were prepared and bonded onto 5-µm silica particles for use as adsorbent in solid-phase microextraction (SPME). Two ILs contained styrene units that allowed for polymerization and higher carbon content of the bonded silica particles. Two polymeric ILs differing by their anion were used to prepare two SPME fibers that were used in both headspace and immersion extractions and compared to commercial fibers. In both sets of experiments, ethyl acetate was used as an internal standard to take into account adsorbent volume differences between the fibers. The polymeric IL fibers are very efficient in headspace SPME for short-chain alcohols. Immersion SPME also can be used with the IL fibers for short-chain alcohols as well as for polar and basic amines that can be extracted at pH 11 without damage to the IL-bonded silica particles. The sensitivities of the two IL fibers differing by the anion were similar. Their efficacy compares favorably to that of commercial fibers for polar analytes. The mechanical strength and durability of the polymeric IL fibers were excellent. FigureChemistry of the polymerized ionic liquid absorbant and its morphology when bonded to the SPME fiber.

Optimization of the Coating Procedure for a High-Throughput 96-Blade Solid Phase Microextraction System Coupled with LC–MS/MS for Analysis of Complex Samples

Biocompatible C18-polyacrylonitrile (PAN) coating was used as the extraction phase for an automated 96-blade solid phase microextraction (SPME) system with thin-film geometry. Three different methods of coating preparation (dipping, brush painting, and spraying) were evaluated; the spraying method was optimum in terms of its stability and reusability. The high-throughput sample preparation was achieved by using a robotic autosampler that enabled simultaneous preparation of 96 samples in 96-well-plate format. The increased volume of the extraction phase of the C18-PAN thin film coating resulted in significant enhancement in the extraction recovery when compared with that of the C18-PAN rod fibers. Various factors, such as reusability, reproducibility, pH stability, and reliability of the coating were evaluated. The results showed that the C18-PAN 96-blade SPME coating presented good extraction recovery, long-term reusability, good reproducibility, and biocompatibility. The limits of detection and quantitation were in the ranges of 0.1-0.3 and 0.5-1 ng/mL for all four analytes.

In vivo solid-phase microextraction for single rodent pharmacokinetics studies of carbamazepine and carbamazepine-10,11-epoxide in mice

The use of solid-phase microextraction (SPME) for in vivo sampling of drugs and metabolites in the bloodstream of freely moving animals eliminates the need for blood withdrawal in order to generate pharmacokinetics (PK) profiles in support of pharmaceutical drug discovery studies. In this study, SPME was applied for in vivo sampling in mice for the first time and enables the use of a single animal to construct the entire PK profile. In vivo SPME sampling procedure used commercial prototype single-use in vivo SPME probes with a biocompatible extractive coating and a polyurethane sampling interface designed to facilitate repeated sampling from the same animal. Pre-equilibrium in vivo SPME sampling, kinetic on-fibre standardization calibration and liquid chromatography-tandem mass spectrometry analysis (LC-MS/MS) were used to determine unbound and total circulating concentrations of carbamazepine (CBZ) and its active metabolite carbamazepine-10,11-epoxide (CBZEP) in mice (n=7) after 2mg/kg intravenous dosing. The method was linear in the range of 1-2000ng/mL CBZ in whole blood with acceptable accuracy (93-97%) and precision (<17% RSD). The single dose PK results obtained using in vivo SPME sampling compare well to results obtained by serial automated blood sampling as well as by the more conventional method of terminal blood collection from multiple animals/time point. In vivo SPME offers the advantages of serial and repeated sampling from the same animal, speed, improved sample clean-up, decreased animal use and the ability to obtain both free and total drug concentrations from the same experiment.

Evaluation of use of a dicationic liquid stationary phase in the fast and conventional gas chromatographic analysis of health-hazardous C18 cis/trans fatty acids.

The present research is focused on the evaluation of one 0.10 mm i.d. and two 0.25 mm i.d., ionic liquid (IL) stationary phase [1,9-di(3-vinyl-imidazolium) nonane bis(trifluoromethyl) sulfonyl imidate] columns, with lengths of 12 (the microbore capillary), 30 and 100 m, in the GC analysis of cis/trans fatty acid methyl esters (FAMEs). The selectivity of the IL columns toward a series of standard C(18:1), C(18:2), and C(18:3) geometric isomers (a group of 22 compounds was subjected to GC analysis) was compared to the performance of a widely used column in the cis/trans FAMEs analysis field, viz., a 100 m x 0.25 mm i.d. capillary with a 0.20 microm stationary phase film of bis-cyanopropyl polysiloxane (SP-2560). The selectivity provided by the IL phase was superior if compared to that of the other well-established capillary. An optimized IL method, using the longer column, was subjected to validation: retention time and peak area intraday precision (n = 5) were good, with RSD values lower than 0.07% and 6.6%, respectively; LODs (considering a S/N of 3) for C(18:1Delta)(9tr) and C(18:2Delta)(9tr,12tr) were 0.15 (7.3 ppm) and 0.18 ng (9.1 ppm) on-column, respectively, while LOQs (considering a S/N of 10) were 0.49 (24.3 ppm) and 0.60 ng (30.2 ppm), respectively; the method was found to be linear, for both trans FAMEs, in the 10-2000 ppm range. For the evaluation of accuracy, a hydrogenated margarine, spiked with known amounts of C(18:3Delta)(9c,12c,15c), was subjected to analysis using C(13:0) as an internal standard.

Innovative Cavitand-Based Sol-Gel Coatings for the Environmental Monitoring of Benzene and Chlorobenzenes via Solid-Phase Microextraction

An innovative and very selective solid-phase microextraction coating synthesized by sol-gel technology was developed for the determination of environmental pollutants such as aromatic hydrocarbons at trace levels in air, water, and soil samples. The obtained fibers, composed of quinoxaline-bridged cavitand units, were characterized in terms of film thickness, morphology, thermal stability, and pH resistance. Fibers, characterized by an average thickness of 56 +/- 6 mum, exhibited an excellent thermal stability until 400 degrees C and a very good fiber-to-fiber and batch-to-batch repeatability with RSD lower than 6%. Finally, the capabilities of the developed coating for the selective sampling of aromatic hydrocarbons were proved, obtaining LOD values in the subnanogram per liter range. Extraction efficiency at least 2-fold higher than that obtained using commercial devices was proved for chlorobenzenes sampling in river water, obtaining extraction recoveries ranging from 87.4 +/- 2.6% to 94.7 +/- 1.9%. The selective desorption of benzene in the presence of high amounts of other airborne pollutants was also demonstrated.

Cavitand-based solid-phase microextraction coating for the selective detection of nitroaromatic explosives in air and soil.

A selective cavitand-based solid-phase microextraction coating was synthesized for the determination of nitroaromatic explosives and explosive taggants at trace levels in air and soil. A quinoxaline cavitand functionalized with a carboxylic group at the upper rim was used to enhance selectivity toward analytes containing nitro groups. The fibers were characterized in terms of film thickness, morphology, thermal stability, and pH resistance. An average coating thickness of 50 (±4) μm, a thermal stability until 400 °C, and an excellent fiber-to-fiber and batch to batch repeatability with RSD lower than 4% were obtained. The capabilities of the developed coating for the selective sampling of nitroaromatic explosives were proved achieving LOD values in the low ppbv and ng kg(-1) range, respectively, for air and soil samples.

Improvement of solid phase microextraction fiber assembly and interface for liquid chromatography

Modifications were made on commercial SPME fiber assembly and SPME-LC interface to improve the applicability of SPME for LC. Polyacrylonitrile (PAN)/C18 bonded fuse silica was used as the fiber coating for LC applications because the fiber coating was not swollen in common LC solvents at room temperature. The inner tubing of SPME fiber assembly was replaced with a 457 μm outside diameter (o.d.) solid nitinol rod. And the coated fiber (o.d. 290 μm) was installed onto the nitinol rod. The inner diameter (i.d.) of the through hole of the ferrule in the SPME-LC interface was enlarged to 508 μm to accommodate the nitinol rod. The much larger inner rod protected the fiber coating from being stripped when the fiber was withdrawn from the SPME-LC interface. The system was evaluated in term of pressure test, desorption optimization, peak shape, carryovers, linear range, precision, and limit of detection (LOD) with polycyclic aromatic hydrocarbons (PAHs) as the test analytes. The results demonstrated that the improved system was robust and reliable. It overcame the drawbacks, such as leak of solvents and damage of fiber coatings, associated with current SPME fibers and SPME-LC interface. Another sealing mechanism was proposed by sealing the nitinol rod with a specially designed poly(ether ether ketone) (PEEK) fitting. The device was fabricated and tested for manual use.

Examination of Selectivities of Thermally Stable Geminal Dicationic Ionic Liquids by Structural Modification

Dicationic ionic liquids (ILs) are widely used as gas chromatography (GC) stationary phases as they show higher thermal stabilities, variety of polarities, and unique selectivities towards certain compounds. An important aspect contributing to them is that they show multiple solvation interactions compared to the traditional GC stationary phases. Dicationic ILs are considered as combination of three structural moieties: (1) cationic head groups; (2) a linkage chain; and (3) the counter anions. Modifications in these structural moieties can alter the chromatographic properties of IL stationary phases. In this study, a series of nine thermally stable IL stationary phases were synthesized by the combination of five different cations, two different linkage chains, and two different anions. Different test mixtures composed of a variety of compounds having different functional groups and polarities were analyzed on these columns. A comparison of the separation patterns of these different compounds on nine different IL columns provided some insights about the effects of structural modifications on the selectivities and polarities of dicationic ILs.