Aqueous Buffer System Research Articles

A metal–organic framework composite loaded with coumarin guests as a turn-on luminescent sensor for detection of cysteine in aqueous solution

In this work, a composite material (ACE@UiO-66-NH2) has been synthesized conveniently by loading a cysteine-responsive molecular probe Acrylic acid 3-acetyl-2-oxo-2H-chromen-7-yl ester (ACE) on a UiO-type zirconium-based metal–organic framework (UiO-66-NH2). Furthermore, the resulting ACE@UiO-66-NH2 demonstrated to be a fluorescent turn-on probe for selective recognition of Cys from other amino acids in aqueous HEPES buffer systems. Additionally, the proposed recognition mechanism has been confirmed by 1H NMR and TLC. The study provided a promising approach to rationally design MOF-based fluorescent sensor for Cys in aqueous system.

Visible Light-Driven CO2 Capture and Release Using Photoactive Pyranine in Water in Continuous Flow.

The urgent need to address climate change and its environmental consequences demands innovative carbon capture technologies, given the relationship between rising global temperatures and increased atmospheric CO2 levels. Here, we present a reversible photochemical carbon capture and release strategy and system utilizing photoactive pyranine in an aqueous bicarbonate buffer system. Control experiments suggested that the photoacid effect occurs at the surface which contributes to CO2 release, complemented by the photothermal effect at the surface and in the bulk. A continuous flow setup employing a tube-in-tube configuration with a hollow fiber membrane demonstrates the efficiency and reliability of the visible light-driven carbon capture system, with the release of CO2 captured from a 15% CO2 feed in the dark, at a rate of 0.48 mmol CO2 per hour to a nitrogen sweep stream under light irradiation at 200 W/m2, a level comparable to solar intensity of visible light (150 W/m2 of blue light -250 W/m2 of blue and green light). The robustness and scalability of the system has been demonstrated, with long-term operation over 7 days yielding 60 mmol (1.34 L CO2 at STP) of cumulated CO2 separation. Additionally, we explored the potential for direct air capture, yielding 3 μmol of CO2 separation over 2 h of operation from a bicarbonate buffer solution saturated with ambient air (420 ppm). This work introduces the prospect of photoswing of carbon capture systems, which can avoid external energy input beyond solar irradiation, offering promising avenues for addressing the challenges associated with climate change.

Electrochemical Studies of Gold in Mops Buffer System

Recent work in this laboratory has investigated interactions of L-cysteine with metal ions.1 For this purpose the MOPS [(3-N-morpholino)propanesulfonic acid] aqueous buffer system has been employed in order to minimize interactions between the metal ions and buffer anions.2 Gold electrodes are expected to be useful in the investigation of L-cysteine and related substances,3 so it is useful to ascertain the background behavior of gold in the aqueous MOPS buffer system. It is the purpose of the present work to determine the voltammetric features in this situation for use in amino acid/metal ion studies.As shown in the accompanying figure, an initial positive-going sweep has a very small oxidation process at +0.45 V vs Ag/AgCl, presumably for the initial oxidation of the gold surface in pH 7.4 MOPS buffer. As this sweep continues, a much larger cathodic process corresponding to bulk gold oxidation is observed. On the return negative-going sweep, there is only a reduction process corresponding to the initial surface oxidation process. It is interesting to note that this reduction process is not observed upon reversing the sweep at only +0.50 V. On the second scan, the initial oxidation process has increased, possibly due to surface roughening in the first scan. Another investigation has involved addition of EDTA to the MOPS solution, resulting in an increase of the second (or bulk) gold oxidation process. This increase is apparently due to the ability of the EDTA to complex the gold cations formed in the oxidation process. This overall behavior is somewhat different compared to that found in other buffer systems such as phosphate,3 and contrasts to such behavior are planned for presentation in this talk. T. Cheek and D. Peña, J. Electrochem. Soc., 167, 155522 (2020).M. H. Ferreira, I. S. S. Pinto, E. V. Soares, and H. M. V. M Soares, RSC Advances, 5(39), 30989 (2015). G.T. Cheek and M.A. Worosz, ECS Trans., 72(27), 1 (2016). doi:10.1149/07227.0001ecst Figure 1

PH dependent physicochemical signatures of amino acids in aqueous buffers: Thermodynamic analysis

The physicochemical signatures such as apparent molar volume (V2,Φ), isoentropic compressibilities (KS,2,φ), and dilution enthalpies of a series of amino acids (AAs) glycine (Gly), L-alanine (Ala), DL-α-amino-n-butyric acid (ABA), L-valine (Val) and L-leucine (Leu) have been measured at various pH values (pH = 2, 5 7 and 9) at a buffer molality of 0.1 mol kg−1 of water at 298.15 K. The data obtained from high sensitivity densimeter and sound analyser along with ultrasensitive isothermal titration calorimetry has been analyzed to evaluate the values of standard partial molar quantities such as for volume at infinite dilution (V2,m0), compressibility KS, 2,m0 and dilution enthalpy (ΔdilH0) of the AAs together with standard partial molar quantities of transfer of these solutes from water to aqueous buffer systems at these pH values. The linear correlation of V2,m0 for this series of AAs against number of carbon atoms in their alkyl chain has been used to evaluate the contribution of zwitterionic end groups (NH3+, COO–), the CH2 group, and other parts of alkyl chains of the AAs to the overall value of V2,m0. The results of standard partial molar transfer properties on volumes, compressibilities and enthalpies of dilution from water to aqueous buffer systems at various pH values have been connected and explained in terms of ion-polar, ion-hydrophilic, hydrophobic-hydrophobic and ion-hydrophobic interactions of the participating groups. These thermodynamic signatures when compared at different pH values suggest that at electrostriction of water dominates at pH 2. Such physicochemical properties of constituents of proteins such as amino acids have not been reported earlier. Thus, the results demonstrate that the interaction of individual amino acid residues with the buffer components does not lead to significant change in their physicochemical properties thereby permitting their use in obtaining mechanistic insights into systems of biological interest consisting of these constituents.

Enhancement of Antioxidant and Anti-Glycation Properties of Beeswax Alcohol in Reconstituted High-Density Lipoprotein: Safeguarding against Carboxymethyllysine Toxicity in Zebrafish.

The antioxidant and anti-inflammatory abilities of beeswax alcohol (BWA) are well reported in animal and human clinical studies, with a significant decrease in malondialdehyde (MDA) in the blood, reduced liver steatosis, and decreased insulin. However, there has been insufficient information to explain BWAs in vitro antioxidant and anti-inflammatory activity owing to its limited solubility in an aqueous buffer system. Herein, three distinct reconstituted high-density lipoproteins (rHDL) were prepared with palmitoyloleoyl phosphatidylcholine (POPC), cholesterol, apolipoprotein A-I (apoA-I), and BWA at molar ratios of 95:5:1:0 (rHDL-0), 95:5:1:0.5 (rHDL-0.5), and 95:5:1:1 (rHDL-1) and examined for antioxidant and anti-glycation effects. A rHDL containing BWA, precisely rHDL-1, displayed a remarkable anti-glycation effect against fructose (final 250 mM), induced glycation of HDL, and prevented proteolytic degradation of apoA-I. Also, BWA incorporated rHDL-0.5, and rHDL-1 displayed substantial antioxidant activity by inhibiting cupric ion-mediated low-density lipoprotein (LDL) oxidation. In contrast to rHDL-0, a 20 and 22% enhancement in ferric ion reduction ability (FRA) and paraoxonase (PON) activity was observed in HDL treated with rHDL-1, signifying the effect of BWA on the antioxidant activity enhancement of HDL. rHDL-1 efficiently inhibits Nε-carboxylmethyllysine (CML)-induced reactive oxygen species (ROS) generation and apoptosis in zebrafish embryos, consequently improving embryo survivability and developmental deformities impaired by the CML. The dermal application of rHDL-1 to the CML-impaired cutaneous wound of the adult zebrafish inhibited ROS production and displayed potent wound-healing activity. Conclusively, incorporating BWA in rHDL significantly enhanced the anti-glycation and antioxidant activities in rHDL via more stabilization of apoA-I with a larger particle size. The rHDL containing BWA facilitated the inherent antioxidant ability of HDL to suppress the CML-induced toxicities in zebrafish embryos and ameliorate CML-aggravated chronic wounds in adult zebrafish.

Efficient Synthesis of Chiral Aryl Alcohol with a Novel Kosakonia radicincitans Isolate in Tween 20/L-carnitine: Lysine-Containing Synergistic Reaction System.

Chiral trifluoromethyl alcohols as vital intermediates are of great interest in fine chemicals and especially in pharmaceutical synthesis. In this work, a novel isolate Kosakonia radicincitans ZJPH202011 was firstly employed as biocatalyst for the synthesis of (R)-1-(4-bromophenyl)-2,2,2-trifluoroethanol ((R)-BPFL) with good enantioselectivity. By optimizing fermentation conditions and bioreduction parameters in aqueous buffer system, the substrate concentration of 1-(4-bromophenyl)-2,2,2-trifluoroethanone (BPFO) was doubled from 10 to 20mM, and the enantiomeric excess (ee) value for (R)-BPFL increased from 88.8 to 96.4%. To improve biocatalytic efficiency by strengthening the mass-transfer rate, natural deep-eutectic solvents, surfactants and cyclodextrins (CDs) were introduced separately in the reaction system as cosolvent. Among them, L-carnitine: lysine (C: Lys, molar ratio 1:2), Tween 20 and γ-CD manifested higher (R)-BPFL yield compared with other same kind of cosolvents. Furthermore, based on the excellent performance of both Tween 20 and C: Lys (1:2) in enhancing BPFO solubility and ameliorating cell permeability, a Tween 20/C: Lys (1:2)-containing integrated reaction system was then established for efficient bioproduction of (R)-BPFL. After optimizing the critical factors involved in BPFO bioreduction in this synergistic reaction system, BPFO loading increased up to 45mM and the yield reached 90.0% within 9h, comparatively only 37.6% yield was acquired in neat aqueous buffer. This is the first report on K. radicincitans cells as new biocatalyst applied in (R)-BPFL preparation, and the developed Tween 20/C: Lys-containing synergistic reaction system has great potential for the synthesis of various chiral alcohols.

Efficient asymmetric synthesis of ethyl (R)-3-hydroxybutyrate by recombinant Escherichia coli cells under high substrate loading using eco-friendly ionic liquids as cosolvent.

Ionic liquids (ILs) which synthesized from bio-renewable materials have recently attracted much attention for their applications in biocatalysis. Ethyl (R)-3-hydroxybutyrate ((R)-EHB) as a versatile chiral intermediate is of great interest in pharmaceutical synthesis. This study focuses on evaluating the performances of choline chloride (ChCl)-based and tetramethylammonium (TMA)-based neoteric ILs in the efficient synthesis of (R)-EHB via the bioreduction of ethyl acetoacetate (EAA) at high substrate loading by recombinant Escherichia coli cells. It was found that choline chloride/glutathione (ChCl/GSH, molar ratio 1:1) and tetramethylammonium/cysteine ([TMA][Cys], molar ratio 1:1) as eco-friendly ILs not only enhanced the solubility of water-insoluble EAA in the aqueous buffer system, but also appropriately improved the membrane permeability of recombinant E. coli cells, thus boosting catalytic reduction efficiency of EAA to (R)-EHB. In the developed ChCl/GSH- or [TMA][Cys]-buffer systems, the space-time yields of (R)-EHB achieved 754.9g/L/d and 726.3g/L/d, respectively, which are much higher than neat aqueous buffer system (537.2g/L/d space-time yield). Meanwhile, positive results have also been demonstrated in the bioreduction of other prochiral ketones in the established IL-buffer systems. This work exhibits an efficient bioprocess for (R)-EHB synthesis under 325g/L (2.5M) substrate loading, and provides promising ChCl/GSH- and [TMA][Cys]-buffer systems employed in the biocatalysis for hydrophobic substrate.

Сорбционно-десорбционные свойства нанокомпозита оксид графена/полигидрохинон при извлечении редкоземельного элемента Sm (III) из уксусно-ацетатных буферных систем

The paper describes the extraction of the rare earth element Sm3+ from aqueous buffer systems by a graphene-based nanostructured composite, modified using an organic polymer – polyhydroquinone. The authors determined the important parameters of the sorption-desorption of Sm3+ ions on a new nanocomposite “graphene oxide/polyhydroquinone” during a batch test, such as: initial concentration, the sorbent weight, pH of the solution, sorption rate constants, maximum sorption capacity of the nanocomposite, percentage sorption and desorption, entropy and enthalpy of the Sm3+ extraction process. The kinetic, isothermal and thermodynamic dependences allowed to propose of the Sm3+ ions adsorption mechanisms. The kinetic data were processed by pseudo-first- and second-order, Elovich and intraparticle diffusion models, and adsorption isotherms — using the Langmuir, Freundlich, Temkin, Dubinin-Radushkevich equations. As a result of kinetic studies, the contact time of the samarium adsorption was determined — 15 min, while the sorption capacity was 100 mg·g–1. It was found that the absorption of Sm3+ ions proceeds by a mixed diffusion mechanism and limited by the interaction “samarium ions : sorbent functional groups”. According to the Langmuir model, the maximum sorbent sorption capacity was 333.3 mg·g–1. Thus, the high efficiency of the developed graphene oxide/polyhydroquinone nanocomposite for purification of aqueous media from rare earth elements was confirmed.

Radical functionalization of thioglycosides in aqueous medium

A visible-light-mediated thiol-ene reaction is successfully operated in aqueous medium. An organic photoredox catalyst (9-mesityl-10-methylacridinum tetrafluoroborate) is used to initiate the radical process to generate thiyl radicals upon light irradiation. Two reaction pathways were discovered in different aqueous buffer systems. A thiol-ene adduct is preferred in acidic reaction medium; while disulfide formation is found to be favored in basic reaction medium. The photocatalytic method is demonstrated to be applicable on unprotected peptides.

Integration of natural deep-eutectic solvent and surfactant for efficient synthesis of chiral aromatic alcohol mediated by Cyberlindnera saturnus whole cells

Efficiency of whole-cell biocatalysis in aqueous buffer solution is mainly limited by the low cellular membrane permeability and poor solubility of hydrophobic non-natural substrate. In this study, highly efficient bioreduction of 3,5-bis(trifluoromethyl)acetophenone (BTAP) to (S)-1-[3,5-bis(trifluoromethyl)phenyl]ethanol ((S)-BTPE) with excellent enantioselectivity (> 99.9 % ee) by a novel yeast isolate Cyberlindnera saturnus ZJPH1807 was achieved via a integrated strategy of natural deep-eutectic solvent (NADES) and surfactant. The introduction of L-carnitine: lysine (C:Lys, molar ratio 1:2) in the reaction medium improved cell membrane permeability effectively and relieved BTAP toxicity to the cells. The Tween-80 addition further accelerated catalytic yield by increasing solubility of BTAP for 7.6-fold compared with neat aqueous buffer solution. After the optimization of key reaction parameters, a 81.0 % yield within 24 h under 500 mM substrate loading was achieved in the established C:Lys/Tween-80-containing system, compared with a 76.4 % yield for 30 h in aqueous buffer system under 200 mM BTAP concentration. This biocatalytic process was also feasible at 500 mL preparation scale with a 79.7 % (S)-BTPE yield under 500 mM BTAP. The developed combinational strategy of NADES with surfactant is an effective approach for enhancing bioreductive efficiency of hydrophobic substrates mediated by whole-cell catalyst, and has great potential in biocatalysis.

An in-depth examination of fatty acid solubility limits in biotherapeutic protein formulations containing polysorbate 20 and polysorbate 80.

Two of the most widely used surfactants to stabilize biologicals against e.g. interfacial stress are polysorbate20 (PS20) and polysorbate 80 (PS80). In recent years, polysorbate degradation in biopharmaceutical formulations has been observed. Polysorbate (PS) is mainly composed of sorbitan and isosorbide fatty acid (FA) esters, varying in their FA composition. Especially hydrolysis, which can be induced chemically as well as enzymatically, leads to the release of FAs from PS. These FAs are poorly soluble in aqueous buffer systems due to their hydrophobic nature and therefore prone to precipitation and particle formation. Since the emergence of particles in liquid formulations has to be avoided, it is important to prevent their formation. This study evaluates the solubility limits of selected FAs, which are likely to be released during the degradation of PS20 and PS80 in the presence of defined PS concentrations. Our results show that the solubility is highly dependent on the pH, the temperature, the used PS concentration and the aliphatic chain of respective FAs. Solubility of FAs, such as palmitic and oleic acid under the conditions determined in this study, are in the range of 3-130µg·ml-1 (12-460µM). Furthermore, the results allow making an estimation to which extent PS may degrade before particle formation in the drug product may be expected.

Development of biocompatible DES/NADES as co-solvents for efficient biosynthesis of chiral alcohols.

The novel deep eutectic solvents (DESs) and natural deep eutectic solvents (NADESs) were designed and synthesized by cell protective components, in which the compounds were derived from natural alternative sources. The performances of designed DESs/NADESs as co-solvent were investigated in asymmetric reduction catalyzed by microbial cells. The DESs/NADESs synthesized by three different types of hydrogen bond receptor (betaine, L-proline and L-carnitine) conferred an advantage over conventional choline chloride-based DESs/NADESs and aqueous buffer system, with regard to efficient bioproduction of (R)-1-[4-(trifluoromethyl)phenyl]ethanol by recombinant Escherichia coli cells. TEM images exhibited that the cell membrane integrity during exposure to the developed NADESs was better than that after treatment with choline chloride-based NADES, which accounted for enhanced catalytic efficiency. This bioprocess was also feasible at 500mL preparation scale with 92.4% yield under 400mM substrate loading. To broaden the applicability of three types of DES/NADESs that increased catalytic efficiency in the process of E. coli-mediated reduction, the production of various chiral alcohols in developed reaction media were further examined, with some positive results. It was also found that lysine-based NADES could even reverse the enantioselectivity of biocatalyst at high water content in the reaction medium. These findings may aid in the development of novel DESs/NADESs for biocatalysis.

A hazard to human health - pesticide residues in some vegetal and animal foodstuff

Highly efficient asymmetric reduction of 2-octanone to (R)-2-octanol catalyzed by immobilized Acetobacter sp. CCTCC M209061 cells was achieved in a biphasic system. Bioreduction conducted in aqueous single phase buffer was limited due to poor solubility and toxicity towards cells cause by product accumulation. Introduction of [C4MIM]·Ac accelerated the biotransformation process, giving 99% yield, >99% product e.e. and 1.42-fold higher initial reaction rate in conversion of 10 mM 2-octanone as substrate, compared with 99% yield, 97.3% product e.e. and 1.57 μmol min−1 of initial reaction rate in a aqueous single phase buffer system containing 6 mM 2-octanone as the optimal substrate concentration. Moreover, in the [C4MIM]·Ac-containing buffer/n-tetradecane biphasic system, the optimal substrate concentration was enhanced by 83 times (500 mM) in comparison with that in aqueous single phase buffer, resulting in 53.4% yield (267 mM), 99% product e.e. with 8.9 mM g-1 h-1 space time yield.

Highly efficient asymmetric reduction of 2-octanone in biphasic system by immobilized Acetobacter sp. CCTCC M209061 cells

Highly efficient asymmetric reduction of 2-octanone to (R)-2-octanol catalyzed by immobilized Acetobacter sp. CCTCC M209061 cells was achieved in a biphasic system. Bioreduction conducted in aqueous single phase buffer was limited due to poor solubility and toxicity towards cells cause by product accumulation. Introduction of [C4MIM]·Ac accelerated the biotransformation process, giving 99% yield, >99% product e.e. and 1.42-fold higher initial reaction rate in conversion of 10 mM 2-octanone as substrate, compared with 99% yield, 97.3% product e.e. and 1.57 μmol min−1 of initial reaction rate in a aqueous single phase buffer system containing 6 mM 2-octanone as the optimal substrate concentration. Moreover, in the [C4MIM]·Ac-containing buffer/n-tetradecane biphasic system, the optimal substrate concentration was enhanced by 83 times (500 mM) in comparison with that in aqueous single phase buffer, resulting in 53.4% yield (267 mM), 99% product e.e. with 8.9 mM g-1 h-1 space time yield.

Indole-based fluorescent sensors for selective sensing of Fe2+ and Fe3+ in aqueous buffer systems and their applications in living cells

New series of indole-based chemosensors; isomeric Iheyamines 5a–5c were synthesized and their fluorescence sensing behaviors toward various metal ions in aqueous buffer systems (90% HEPES buffered/MeOH, pH 7.2) were explored. The results revealed that isomeric Iheyamine 5a–5c could act as Fe2+ and Fe3+ chemosensors, in which their sensitivity and selectivity for Fe2+ and Fe3+ sensing depended on their substituent on benzene ring at position 5. The isomeric Iheyamine 5c containing electron withdrawing group exhibited nonselective toward Fe2+ and Fe3+ and its sensitivity toward Fe2+ and Fe3+ was much lower than those of 5a and 5b. For the isomeric Iheyamine 5a, it exhibited “turn-off” fluorescent response upon the addition of Fe2+ and Fe3+ with the discrimination of other metal ions such as Hg2+, Cu2+, Zn2+, Al3+, Ag+, Pb2+, Cr2+, Ni2+, Mg2+, K+, Ca2+, Li+, Co2+, Na+, Ba2+, Cd2+ and Mn2+. Moreover, although the isomeric Iheyamine 5b showed higher sensitivity than 5a, the Fe2+ and Fe3+ binding of 5b could be interfered by Cu2+. In addition, the Fe2+ could be completely oxidized to Fe3+ by simple H2O2 oxidation, consequently, the total concentration of Fe could be determined. The detection limits of isomeric Iheyamines 5a and 5b for Fe3+ were found to be 0.12 ppm and 0.03 ppm, respectively, which were lower than the maximum allowed level of Fe3+ in drinking water specified U.S. Environmental Protection Agency (U.S.EPA) and World Health Organization (WHO). In addition, isomeric Iheyamine 5b showed the potential for the detection of Fe3+ in HeLa cellular system and could be the good candidate for Fe3+ sensor used in biological system.

Nonionic surfactants and their effects on asymmetric reduction of 2-octanone with Saccharomyces cerevisiae

In an aqueous buffer system, serious reverse and side reactions were found in the asymmetric reduction of 2-octanone with Saccharomyces cerevisiae. However, some nonionic surfactants added to the aqueous buffer system improved the bioreduction process by decreasing the reverse and side reaction rates in addition to effectively increasing the average positive reaction rate. Further, a shorter carbon chain length of hydrophilic or hydrophobic moieties in surfactants resulted in a higher yield of (S)-2-octanol. The alkylphenol ethoxylate surfactants had a less influence than polyoxyethylenesorbitan trialiphatic surfactants on the product e.e. It suggested that the product e.e. resulting from the change of carbon chain length of the hydrophobic moieties varied markedly compared with the change of carbon chain length of the hydrophilic moiety. Emulsifier OP-10 and Tween 20 markedly enhanced the yield and product e.e. at the concentration of 0.4 mmol L−1 with a yield of 73.3 and 93.2%, and the product e.e. of 99.2 and 99.3%, respectively, at the reaction time of 96 h.

Dissolution of commercial nanoscale silica particles in electrolyte solution: The importance of the solid-solvent-ratio to physical and chemical properties of the solid-liquid interface

Dissolution experiments of silica nanoparticles (NPs) were performed in an aqueous TRIS buffer system (pH=7.6) at 25°C and the effect of the particles to solvent ratio (PSR) was studied. Increasing the PSR distinctly promotes the kinetic size effect described previously, which leads to solute concentrations exceeding the thermodynamic saturation concentration under the respective conditions considerably. The striking experimental observations readily are rationalized by analysis of the Gibbs free energy (GFE) of the system. In the extended model approach the dissolution and Ostwald ripening is described using gradient curves of GFE, which have been calculated considering a variable interfacial tension of the solid-liquid-interface, γ. The GFE analysis provides size, RP, and concentration, ZP, of NPs as a function of dissolution time. The variation of the interfacial tension with particle size, γ(RP), is a consequence of the properties of the electric double layer, surface charge and surface potential, around the NPs. Suggestions are made how to predict the dissolution process from properties of the initial system solid-solution. Any meaningful report of dissolution experiments should include the saturation concentration of the material that forms the NPs as thermodynamic criterion, and the time necessary to obtain half of the saturation concentration in the system considered as kinetic criterion. Both aforementioned criteria are accessible from experiments without any model considerations.

Fast Protein Liquid Chromatography.

Fast protein liquid chromatography (FPLC) is a form of high-performance chromatography that takes advantage of the high resolution made possible by small-diameter stationary phases. It was originally developed for proteins and features high loading capacity, biocompatible aqueous buffer systems, fast flow rates, and availability of stationary phases in most common chromatography modes (e.g., ion exchange, gel filtration, reversed phase, and affinity). The system makes reproducible separation possible by incorporating a high level of automation including autosamplers, gradient program control, and peak collection. In addition to proteins, the method is applicable to other kinds of biological samples including oligonucleotides and plasmids. The most common type of FPLC experiment is anion exchange of proteins. This chapter describes such an experiment carried out using an ÄKTA FPLC explorer system (Amersham Pharmacia Biotech, Sweden).

Anilide formation from thioacids and perfluoroaryl azides.

A metal-free method for fast and clean anilide formation from perfluoroaryl azide and thioacid is presented. The reaction proved highly efficient, displaying fast kinetics, high yield, and good chemoselectivity. The transformation was compatible with various solvents and tolerant to a wide variety of functional groups, and it showed high performance in polar protic/aprotic media, including aqueous buffer systems.

Fast and efficient extraction of DNA from meat and meat derived products using aqueous ionic liquid buffer systems

We present a fast and efficient strategy for DNA extraction from meat based on aqueous-ionic liquid systems that could extract DNA in significantly higher yields compared to the pure phosphate buffer.