Physicochemical Properties of 20 Ionic Liquids Prepared by the Carbonate-Based IL (CBILS) Process.

Ionic liquids (ILs) are an emerging materials' class with applications in areas such as energy storage, catalysis, and biomass dissolution and processing. Their physicochemical properties including surface tension, viscosity, density and their interplay between cation and anion chemistry are decisive in these applications. For many commercially available ILs, a full set of physicochemical data is not available. Here, we extend the knowledge base by providing physicochemical properties such as density (20 and 25 °C), refractive index (20 and 25 °C), surface tension (23 °C, including polar and dispersive components), and shear viscosity (ambient atmosphere, shear rate 1-200 s-1), for 20 commercial ILs. A correlation between the crystal volume, dispersive surface tension, and shear viscosity is introduced as a predictive tool, allowing for viscosity estimation. Systematic exploration of cation/anion alkyl side chain lengths reveals the impact on the IL's physicochemical attributes. Increasing the anion's headgroup decreases surface tension up to 35.7% and consequently shear viscosity. We further demonstrate that the dispersive part of the surface tension linearly correlates with the refractive index of the ionic liquid. While we provide additional physicochemical data, the screening and modeling efforts will contribute to better structure property predictions enabling faster progress in design and applications of ILs.

Design and evaluation of improved magnetic stir bars for single-mode microwave reactors.

Magnetic stirring in sealed cylindrical vessels designed for use in single-mode microwave instruments is typically less than optimal, and is not comparable to the efficient agitation that can be generally obtained in a round-bottomed flask fitted with a suitable magnetic stir bar or using overhead mechanical stirring systems. A new "vertical blade" stir bar design that improves the stirring performance in the very narrow, flow-constricting microwave vessels has been developed and evaluated for several different transformations where stirring and efficient agitation are known to be of importance. The better performance of these novel stirrers compared to the traditional cylindrical stir bar design is not only due to the geometry of the stirrer but also to the utilization of a magnetic material with a stronger magnetic transmission force (Sm2Co17) compared to standard ferrite or AlNiCo alloys. For all three tested cases involving solid/liquid, liquid/liquid and highly viscous reaction systems, the new vertical blade stirrers showed a distinctively improved performance resulting in higher conversions and/or product yields.

Parallel microwave chemistry in silicon carbide microtiter platforms: a review.

In this review, applications of silicon carbide-based microtiter platforms designed for use in combination with dedicated multimode microwave reactors are described. These platforms are employed not only for the efficient parallel synthesis of compound libraries, but also in the context of high-throughput reaction screening/optimization and a number of other (bio)analytical and biomedical applications. Since the semiconducting plate material (silicon carbide) is strongly microwave absorbing and possesses high thermal conductivity, no temperature gradients across the microtiter plate exist. Therefore, many of the disadvantages experienced in attempting to perform microtiter plate chemistry under conventional microwave conditions can be eliminated. In general, the silicon carbide-based microtiter platforms allow sealed vessel processing (either directly in the well or in glass vials placed into the wells) of volumes ranging from 0.02-3.0 mL at a maximum temperature/pressure limit of 200°C/20 bar. Depending on the specific plate and rotor configuration, a maximum of 80-192 transformations can be carried out in parallel in a single microwave irradiation experiment under strict temperature control. A platform type utilizing HPLC/GC vials as reaction vessels allows analysis directly from the reaction vessel eliminating the need for a transfer step from the reaction to the analysis vial. The latter system is particularly useful for analytical applications as well as reaction optimization/screening.

Ganglioside lipidomics of CNS myelination using direct infusion shotgun mass spectrometry.

Gangliosides are present and concentrated in axons and implicated in axon-myelin interactions, but how ganglioside composition changes during myelin formation is not known. Here, we present a direct infusion (shotgun) lipidomics method to analyze gangliosides in small amounts of tissue reproducibly and with high sensitivity. We resolve the mouse ganglioside lipidome during development and adulthood and determine the ganglioside content of mice lacking the St3gal5 and B4galnt1 genes that synthesize most ganglioside species. Our results reveal substantial changes in the ganglioside lipidome during the formation of myelinated nerve fibers. In sum, we provide insights into the CNS ganglioside lipidome with a quantitative and sensitive mass spectrometry method. Since this method is compatible with global lipidomic profiling, it will provide insights into ganglioside function in physiology and pathology.

Microwave-assisted derivatization procedures for gas chromatography/mass spectrometry analysis.

In this review, published applications of microwave-assisted derivatization procedures for gas chromatography/mass spectrometry (GC/MS) are summarized. Among the broad range of analytical techniques available, GC/MS is still the method of choice for most high-throughput screening procedures in forensic/clinical toxicology, doping control and food and environmental analysis. Despite the many advantages of the GC/MS method, time-consuming derivatization steps are often required in order to obtain desirable chromatographic characteristics or to improve the stability and detectability of the target analytes. These derivatization processes typically require reaction times from 30 min up to several hours at elevated temperature. In contrast, microwave protocols have demonstrated to be able to reduce the time required for derivatization to a few minutes, and can thus very effectively shorten the overall analysis time, in particular when carried out in a high-throughput format. Herein, the literature in this field is summarized and recent experimental techniques for performing parallel GC/MS derivatization protocols are discussed.

Plasma lipidomics of monozygotic twins discordant for multiple sclerosis.

Blood biomarkers of multiple sclerosis (MS) can provide a better understanding of pathophysiology and enable disease monitoring. Here, we performed quantitative shotgun lipidomics on the plasma of a unique cohort of 73 monozygotic twins discordant for MS. We analyzed 243 lipid species, evaluated lipid features such as fatty acyl chain length and number of acyl chain double bonds, and detected phospholipids that were significantly altered in the plasma of co-twins with MS compared to their non-affected siblings. Strikingly, changes were most prominent in ether phosphatidylethanolamines and ether phosphatidylcholines, suggesting a role for altered lipid signaling in the disease.

Shotgun Lipidomics Discovered Diurnal Regulation of Lipid Metabolism Linked to Insulin Sensitivity in Nondiabetic Men.

CONTEXT: Meal timing affects metabolic homeostasis and body weight, but how composition and timing of meals affect plasma lipidomics in humans is not well studied. OBJECTIVE: We used high throughput shotgun plasma lipidomics to investigate effects of timing of carbohydrate and fat intake on lipid metabolism and its relation to glycemic control. DESIGN: 29 nondiabetic men consumed (1) a high-carb test meal (MTT-HC) at 09.00 and a high-fat meal (MTT-HF) at 15.40; or (2) MTT-HF at 09.00 and MTT-HC at 15.40. Blood was sampled before and 180 minutes after completion of each MTT. Subcutaneous adipose tissue (SAT) was collected after overnight fast and both MTTs. Prior to each investigation day, participants consumed a 4-week isocaloric diet of the same composition: (1) high-carb meals until 13.30 and high-fat meals between 16.30 and 22:00 or (2) the inverse order. RESULTS: 12 hour daily lipid patterns showed a complex regulation by both the time of day (67.8%) and meal composition (55.4%). A third of lipids showed a diurnal variation in postprandial responses to the same meal with mostly higher responses in the morning than in the afternoon. Triacylglycerols containing shorter and more saturated fatty acids were enriched in the morning. SAT transcripts involved in fatty acid synthesis and desaturation showed no diurnal variation. Diurnal changes of 7 lipid classes were negatively associated with insulin sensitivity, but not with glucose and insulin response or insulin secretion. CONCLUSIONS: This study identified postprandial plasma lipid profiles as being strongly affected by meal timing and associated with insulin sensitivity.

High-throughput experimentation platform: parallel microwave chemistry in HPLC/GC vials.

A high-throughput reaction platform for performing parallel microwave chemistry in sealed HPLC/GC vials is described. The system consists of a strongly microwave-absorbing silicon carbide plate with 20 cylindrical wells of appropriate dimensions to be fitted with standard HPLC/GC autosampler vials serving as reaction vessels. In combination with an aluminum sealing plate the setup can be used for microwave processing reaction volumes from 0.5-1.5 mL at a maximum temperature/pressure limit of 250 degrees C/20 bar. The parallel reaction platform displays excellent temperature and reaction homogeneity and has been used for high-throughput reaction optimization studies involving the parallel screening of catalyst, solvent and substrate reactivity for esterification reactions and metal-catalyzed dehydrative C-C couplings.

Parallel microwave synthesis of 2-styrylquinazolin-4(3H)-ones in a high-throughput platform using HPLC/GC vials as reaction vessels.

The application of a high-throughput reaction platform for performing parallel microwave synthesis in sealed HPLC/GC vials contained in a strongly microwave-absorbing silicon carbide plate is described. The use of aluminum crimp caps with PTFE coated silicone septa in combination with an appropriate plate sealing mechanism allows processing of reaction volumes from 0.5-1.5 mL at temperatures of approximately 250 degrees C and pressures of up to approximately 20 bar. A library of 39 2-styrylquinazolin-4(3H)-one derivatives was prepared in a two-step/one-pot parallel fashion involving the initial three-component condensation of four anthranilic acids with acetic anhydride and ammonium acetate at 250 degrees C for 30 min. This was followed by catalyst-free condensation of the resulting 2-methylquinazolinones with a selection of 15 aromatic aldehydes. Compared to single-mode sequential microwave synthesis, the overall processing times for library synthesis could be significantly reduced.

Parallel microwave chemistry in silicon carbide reactor platforms: an in-depth investigation into heating characteristics.

The heating behavior of silicon carbide reaction platforms under 2.45 GHz microwave irradiation was investigated with the aid of online thermoimaging cameras and multiple-channel fiber-optic probe temperature sensors placed inside the wells/vials of the silicon carbide microtiter plates. Microwave irradiation leads to a rapid and homogeneous heating of the entire plate, with minimal deviations in the temperature recorded at different positions of the plate or inside the wells. In temperature-controlled experiments using dedicated multimode reactors, solvents with different microwave absorption characteristics can be heated in parallel in individual wells/vials of the silicon carbide plate reaching the same set temperature. Due to the large heat capacity and high thermal conductivity of silicon carbide, the plates are able to moderate any field inhomogeneities inside a microwave cavity. Although the heating of the plates can be performed extremely efficiently inside a microwave reactor, heating and synthetic applications can alternatively be carried out by applying conventional conductive heating of the silicon carbide plates on a standard hotplate. Due to the slower heating of the silicon carbide material under these conditions, somewhat longer reaction times will be required.

Continuous-flow synthesis of adipic acid from cyclohexene using hydrogen peroxide in high-temperature explosive regimes.

Safe only in a microreactor! The synthesis of adipic acid from cyclohexene by tungstic acid-catalyzed oxidation using hydrogen peroxide following the classical Noyori protocol can be accomplished in good yields with residence times as short as 20 min at 140 °C using a safe and scalable microreactor environment. Under these intensified conditions the use of a phase-transfer catalyst is not required.

Can electromagnetic fields influence the structure and enzymatic digest of proteins? A critical evaluation of microwave-assisted proteomics protocols.

This study reevaluates the putative advantages of microwave-assisted tryptic digests compared to conventionally heated protocols performed at the same temperature. An initial investigation of enzyme stability in a temperature range of 37-80 °C demonstrated that trypsin activity declines sharply at temperatures above 60 °C, regardless if microwave dielectric heating or conventional heating is employed. Tryptic digests of three proteins of different size (bovine serum albumin, cytochrome c and ß-casein) were thus performed at 37 °C and 50 °C using both microwave and conventional heating applying accurate internal fiber-optic probe reaction temperature measurements. The impact of the heating method on protein degradation and peptide fragment generation was analyzed by SDS-PAGE and MALDI-TOF-MS. Time-dependent tryptic digestion of the three proteins and subsequent analysis of the corresponding cleavage products by MALDI-TOF provided virtually identical results for both microwave and conventional heating. In addition, the impact of electromagnetic field strength on the tertiary structure of trypsin and BSA was evaluated by molecular mechanics calculations. These simulations revealed that the applied field in a typical laboratory microwave reactor is 3-4 orders of magnitude too low to induce conformational changes in proteins or enzymes.

A high-throughput platform for low-volume high-temperature/pressure sealed vessel solvent extractions.

A high-throughput platform for performing parallel solvent extractions in sealed HPLC/GC vials inside a microwave reactor is described. The system consist of a strongly microwave-absorbing silicon carbide plate with 20 cylindrical wells of appropriate dimensions to be fitted with standard HPLC/GC autosampler vials serving as extraction vessels. Due to the possibility of heating up to four heating platforms simultaneously (80 vials), efficient parallel analytical-scale solvent extractions can be performed using volumes of 0.5-1.5 mL at a maximum temperature/pressure limit of 200°C/20 bar. Since the extraction and subsequent analysis by either gas chromatography or liquid chromatography coupled with mass detection (GC-MS or LC-MS) is performed directly from the autosampler vial, errors caused by sample transfer can be minimized. The platform was evaluated for the extraction and quantification of caffeine from commercial coffee powders assessing different solvent types, extraction temperatures and times. For example, 141±11 µg caffeine (5 mg coffee powder) were extracted during a single extraction cycle using methanol as extraction solvent, whereas only 90±11 were obtained performing the extraction in methylene chloride, applying the same reaction conditions (90°C, 10 min). In multiple extraction experiments a total of ~150 µg caffeine was extracted from 5 mg commercial coffee powder. In addition to the quantitative caffeine determination, a comparative qualitative analysis of the liquid phase coffee extracts and the headspace volatiles was performed, placing special emphasis on headspace analysis using solid-phase microextraction (SPME) techniques. The miniaturized parallel extraction technique introduced herein allows solvent extractions to be performed at significantly expanded temperature/pressure limits and shortened extraction times, using standard HPLC autosampler vials as reaction vessels. Remarkable differences regarding peak pattern and main peaks were observed when low-temperature extraction (60°C) and high-temperature extraction (160°C) are compared prior to headspace-SPME-GC-MS performed in the same HPLC/GC vials.

Microwave-assisted forced degradation using high-throughput microtiter platforms.

Parallel microwave-assisted forced degradation in sealed HPLC/GC vials utilizing a high-throughput platform is described. The platform is made out of strongly microwave absorbing silicon carbide (SiC) plates providing 20 bore holes having the appropriate dimensions to be fitted with standard autosampler HPLC/GC vials serving as reaction vessels. Due to the possibility of heating up to four SiC platforms simultaneously (80 reactions) in a dedicated multimode microwave cavity with online temperature control, efficient parallel forced degradation studies can be performed at temperatures and pressures of up to 200 °C and 20 bar, respectively. Since degradation reactions and analyses are performed in the same vessel, the sample handling effort is reduced and errors caused by a required transfer step are avoided. As proof-of-concept, the platform was evaluated for the parallel testing of various stress conditions on the drug indomethacin. The obtained data provided a rapid overview over suitable stress conditions at high temperatures, implicating a significant reduction in time required for the forced degradation compared to conventional methods at room temperature. Applying acidic (0.01-0.1 M HCl, 1-15 M AcOH), basic (0.001-0.01 M NaOH, 0.001-0.01 M NaHCO(3)) and oxidative (0.001-0.02% H(2)O(2)) stress conditions at 150 °C for 5 min resulted in similar indomethacin degradation levels requiring 0.5-20 h at lower temperatures (25-100 °C). In addition solvent stability tests exposing indomethacin to 20 different, mostly organic, solvents at 150 °C and 160 °C for 30 min and the exposure of the solid drug to various gases (N(2), Ar, O(2), NH(3), air), applying high temperatures are presented.

A miniaturized microtiter plate protocol for the determination of in selenized yeast via enzymatic hydrolysis of protein-bound.

This paper describes a simple/low volume enzymatic extraction method for selenomethionine (SeMet) determination in selenized yeast samples. In contrast to traditional methods which generally utilize large sample volumes consuming significant amounts of costly enzymes, the modified protocol employs a microtiter plate format allowing a reduction of the required sample volumes to 1 mL per extract. The extraction is performed in a parallel (5 × 4 = 20 position microtiter plate) reaction platform made out of sintered silicon carbide, fitted with standard disposable glass HPLC/GC vials. Due to the high thermal conductivity of silicon carbide, this set-up can be placed on a standard hotplate to accurately maintain the desired extraction conditions (37 °C, 20 h) for all positions of the microtiter plate. Hydrolysis of selenium-enriched yeast with a combination of protease XIV and lipase VII (ratio 2 : 1, w/w) using these low-volume conditions provided identical results to the more traditional high-volume method. The amount of SeMet was determined by HPLC/ICPMS and confirmed a high recovery rate for SeMet (93 ± 2%, n = 3) for the certified reference material SELM-1.

Microwave-assisted high-throughput derivatization techniques utilizing silicon carbide microtiter platforms.

Parallel microwave-assisted gas chromatography (GC) derivatization protocols utilizing a silicon carbide (SiC)-based microtiter plate platform fitted with screw-capped GC vials were developed. For three selected standard derivatization protocols such as acetylation (exemplified for morphine), pentafluoropropionylation (for 6-monoacetylmorphine) and trimethylsilylation (for Delta(9)-tetrahydrocannabinol) complete derivatization was achieved within 5min at 100 degrees C in a dedicated multimode microwave instrument using online temperature monitoring. Microwave irradiation leads to rapid and homogeneous heating of the strongly microwave-absorbing SiC plate, with minimal deviations in the temperature recorded at different positions of the plate. The current platform allows the simultaneous derivatization of 80 reaction mixtures under strictly controlled temperature conditions. Similar results can also be obtained using a standard hotplate as heating source, although heating to the target temperature of 100 degrees C is slightly slower. The results demonstrate that parallel microwave derivatization procedures can significantly reduce the overall analysis time and increase sample throughput for GC-MS-based analytical methods.

Microwave-assisted high-throughput acid hydrolysis in silicon carbide microtiter platforms--a rapid and low volume sample preparation technique for total amino acid analysis in proteins and peptides.

An efficient microwave-assisted high-throughput protein hydrolysis protocol was developed utilizing strongly microwave absorbing silicon carbide-based microtiter platforms. The plates are equipped with 20 bore holes having the proper dimensions for holding standard screw-capped HPLC/GC vials. Due to the possibility of heating up to four heating platforms simultaneously (80 vials), parallel microwave-assisted acid hydrolyses can be performed under carefully controlled conditions significantly reducing the overall time required for protein hydrolysis and the subsequent evaporation step required for larger volumes of acid. An extensive optimization of the hydrolysis conditions has demonstrated that 5min irradiation at 160°C with 6N HCl leads to comparable results in terms of total and individual amino acid recovery as the traditional method requiring 24h heating at 110°C. Complete hydrolysis of several proteins and synthetic peptides was performed using 25µg of sample material and 100µL of 6N HCl in a dedicated low-volume HPLC/GC vial. Since the hydrolysis and subsequent analysis can be performed from the same vial, errors caused by sample transfer can be minimized. Control experiments have demonstrated that the observed rate enhancements are the result of a purely thermal/kinetic effect as a consequence of the considerable higher reaction temperatures.

An evaluation of microwave-assisted derivatization procedures using hyphenated mass spectrometric techniques.

The potential of microwave-assisted derivatization techniques in systematic toxicological analysis using gas chromatography coupled with mass spectrometry (GC-MS) was evaluated. Special emphasis was placed on the use of dedicated microwave reactors incorporating online temperature and pressure control. The use of such equipment allowed a detailed analysis of several microwave-assisted derivatization protocols comparing the efficiency of microwave and conventional heating methods utilizing a combination of GC-MS and liquid chromatography coupled with mass detection (LC-MS and LC-MS/MS) techniques. These studies revealed that for standard derivatization protocols such as acetylation (exemplified for codeine and morphine), pentafluoropropionylation (for 6-monoacetylmorphine) and trimethylsilylation (for Delta9-tetrahydrocannabinol) a reaction time of 5 min at 100 degrees C in a microwave reactor was sufficient to allow for an effective derivatization. Control experiments using standard operating procedures (30 min at 60 degrees C conventional heating) indicated that the faster derivatization under microwave irradiation is a consequence of the higher reaction temperatures that can rapidly be attained in a sealed vessel and the more efficient heat transfer to the reaction mixture applying direct in core microwave dielectric heating. The results suggest that microwave derivatization procedures can significantly reduce the overall analysis time and increase sample throughput for GC-MS-based analytical methods.