Enzymatic Extraction of Bioactive and Self‐Assembling Wool Keratin for Biomedical Applications

Keratin is widely recognized as a high‐quality renewable protein resource for biomedical applications. Despite their extensive existence, keratin resources such as feathers, wool, and hair exhibit high stability and mechanical properties because of their high disulfide bond content. Consequently, keratin extraction is challenging and its application is greatly hindered. In this work, a biological extraction strategy is proposed for the preparation of bioactive keratin and the fabrication of self‐assembled keratin hydrogels (KHs). Based on moderate and controlled hydrolysis by keratinase, keratin with a high molecular weight of approximately 45 and 28 kDa that retain its intrinsic bioactivities is obtained. The keratin products show excellent ability to promote cell growth and migration and are conferred with significant antioxidant ability because of their intrinsically high cysteine content. In addition, without the presence of any cross‐linking agent, the extracted keratin can self‐assemble into injectable hydrogels. The KHs exhibit a porous network structure and 3D culture ability, showing potential in promoting wound healing. This enzyme‐driven keratin extraction strategy opens up a new approach for the preparation of keratin that can self‐assemble into injectable hydrogels for biomedical engineering.     

Determination of terbinafine in human plasma using UPLC–MS/MS: Application to a bioequivalence study in healthy subjects

A high‐throughput and sensitive ultra‐performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) method has been developed for the determination of terbinafine in human plasma. The method employed liquid–liquid extraction of terbinafine and terbinafine‐d7 (used as internal standard) from 100 μL human plasma with ethyl acetate–n‐hexane (80:20, v/v) solvent mixture. Chromatography was performed on a BEH C₁₈ (50 × 2.1 mm, 1.7 μm) column using acetonitrile–8.0 mm ammonium formate, pH 3.5 (85:15, v/v) under isocratic elution. For quantitative analysis, MS/MS ion transitions were monitored at m/z 292.2/141.1 and m/z 299.1/148.2 for terbinafine and terbinafine‐d7, respectively, using electrospray ionization in the positive mode. The method was validated according to regulatory guidance for selectivity, sensitivity, linearity, recovery, matrix effect, stability, dilution reliability and ruggedness with acceptable accuracy and precision. The method shows good linearity over the tested concentration range from 1.00 to 2000 ng/mL (r² ≥ 0.9984). The intra‐batch and inter‐batch precision (CV) was 1.8–3.2 and 2.1–4.5%, respectively. The method was successfully applied to a bioequivalence study with 250 mg terbinafine in 32 healthy subjects. The major advantage of this method includes higher sensitivity, small plasma volume for processing and a short analysis time.     

Modified layered bimetallic compound used as a novel adsorbent for the solid-phase extraction of monoaromatic hydrocarbons from water samples prior to gas chromatography–mass spectrometry

This study reports a promising method of solid-phase extraction for determining the toluene, ethylbenzene, p-xylene, m-xylene, o-xylene, 1,3,5-trimethylbenzene, and 1,2,4-trimethylbenzene in water samples by gas chromatography–mass spectrometry (GC–MS). Prior to this procedure, the magnesium–aluminum bimetallic hydroxides modified with sodium dodecylbenzenesulfonate (Mg/Al-SDBS-LDH) were prepared and served as the novel solid-phase extractant. The Mg/Al-SDBS-LDH has advantage of good hydrophobicity and larger spacing which facilitates the monoaromatic hydrocarbons (MAHCs) into the interlayer for adsorption. As a result, the seven MAHCs in 500 mL water samples were enriched greatly, and the theoretical enrichment factor reached to 125 times. Under the optimized conditions of solid-phase extraction (SPE) and GC–MS, the mass concentration of each MAHC (0.005–10, 0.01–10, or 0.05–10 ng/mL) had a fine linear relationship with peak area. The correlation coefficients were more than 0.995. The detection limits were between 0.001 and 0.01 ng/mL, and the RSD were between 3.1% and 6.6%. The method had been applied to determine the seven MAHCs in the Dongfengqu river water and laboratory wastewater of Chengdu University of Technology successfully.     

New insights into the extraction of uranium(VI) by an N,N-dialkylamide

This paper presents the methodology developed in order to thoroughly characterise a solvent extraction system containing high solute concentrations. The chemical system selected is N,N-(2-ethylhexyl)isobutyramide (DEHiBA) diluted in one alkane with increasing concentration of uranium(VI). Combining experiments with theoretical calculations allowed a deeper understanding of the extraction mechanism. A thermodynamic study was performed by the classical van't Hoff method and also by direct calorimetry to provide the enthalpies of extraction and specific heats. Dedicated methods like vapour pressure osmometry and electrospray ionisation mass spectrometry analysis provide information about the stoichiometry of the extracted species. Spectroscopic investigations with ultraviolet–visible and Fourier transform infrared probed the uranium coordination. Finally, a combination of molecular dynamics simulations, and small and wide-angle X-ray scattering experiments investigated the organisation in the organic phase beyond the molecular scale. It was shown that the high concentrations of uranium extracted have no influence on the stoichiometry of the complexes and the coordination of uranium in the inner sphere. The thermodynamic properties related to the extraction process and obtained with a fine consideration of the activity coefficients showed to be the same as those found with trace concentration. However, an unexpected organisation beyond the molecular scale was observed with an important role of nitrates as bridging ligands which could explain some physico-chemical properties. This approach could be applied to other chemical systems (other N,N-dialkylamides or other cations) to identify the origin of the different affinities between ligands and the difference of selectivity between cations.     

A modified sequential extraction method for arsenic fractionation in sediments

A modified sequential extraction method was developed to characterize arsenic (As) associated with different solid constituents in surficial deposits (sediments), which are unconsolidated glacial deposits overlying bedrock. Current sequential extraction methods produce a significant amount of unresolved As in the residual fraction, but our proposed scheme can fractionate >90% of the As present in sediments. Sediment samples containing different As concentrations (3–35 μg g⁻¹) were used to assess the developed method. The pooled amount of As recovered from all the fractions using the developed method was similar (83–122%) to the total As extracted by acid digestion. The concentrations of As in different fractions using the developed scheme were comparable (89–106%) to the As fractions obtained by other existing methods. The developed method was also evaluated for the sequential extraction of other metals such as copper (Cu), cobalt (Co), chromium (Cr) and strontium (Sr) in the sediment samples. The pooled concentrations of these four individual metals from all the fractions were similar (96–104%) to their total concentrations extracted by acid digestion. During method development, we used extractants that did not contain chloride to eliminate formation of polyatomic ions of argon chloride (⁴⁰Ar³⁵Cl) that interfered with ⁷⁵As when analyzed using inductively coupled plasma mass spectrometer (ICP-MS). The results suggest that the developed method can reliably be employed for complete As and other metals’ fractionation in sediments using ICP-MS.     

A novel molecularly imprinted method with computational simulation for the affinity isolation and knockout of baicalein from Scutellaria baicalensis

A novel molecularly imprinted polymer (MIP) was synthesized by precipitation polymerization with baicalein (BAI) as the template and used as solid‐phase extraction (SPE) adsorbent, aiming at the affinity isolation and selective knockout of BAI from Scutellaria baicalensis Georgi (SB). We used computational simulation to predict the optimal functional monomer, polymerization solvent and molar ratio of template to functional monomer. Characterization and performance tests revealed that MIP exhibited uniform spherical morphology, rapid binding kinetics, and higher adsorption capacity for BAI compared with nonimprinted polymer (NIP). The application of MIP in SPE coupled with high‐performance liquid chromatography to extract BAI from SB showed excellent recovery (94.3%) and purity (97.0%). Not only the single BAI compound, but also the BAI‐removed SB extract was obtained by one‐step process. This new method is useful for isolation and knockout of key bioactive compounds from herbal medicines. Copyright © 2015 John Wiley & Sons, Ltd.     

Saccharide Recognition by a Three-Arm-Shaped Host Having Preorganized Three-Dimensional Hydrogen-Bonding Sites

Generally, cage-shaped hosts for saccharides can bind strongly to guest molecules because of the three-dimensional preorganized hydrogen-bonding sites. However, the preparation of cage molecules is often difficult because of the low yield of the macrocyclization step. Here, we report a three-arm-shaped molecule possessing pyridine-acetylene-phenol units as a new kind of host having a preorganized three-dimensional hydrogen-bonding site. This three-arm-shaped host was readily prepared compared to a cage-shaped analogue. This host associated with lipophilic glycosides to form chiral complexes, and the association constants were sufficiently high as to be comparable to those of the cage-shaped analogue. Furthermore, this host extracted native monosaccharides into a lipophilic solvent.     

Automated capillary electrophoresis with on-line preconcentration by solid phase extraction using a sequential injection manifold and contactless conductivity detection

An extension of a capillary electrophoresis instrument coupled to a sequential injection analysis manifold was developed for automated measurements with on-line solid-phase extraction preconcentration. An in-house built capacitively coupled contactless conductivity detector was employed for sensitive detection with narrow capillaries of 25 μm internal diameter. The system was assembled into standardized 19 in. frames and racks for easy transport and mobile deployment. The system can be left running unattendedly without manual intervention with good operation stability. To demonstrate the application of the system, a method for the determination of four drugs, namely ibuprofen, diclofenac, naproxen and bezafibrate, was developed with enrichment factors of up to several hundreds. Detection of the drug residues down to the nM-scale was found possible and the method was found suitable for the detection of ibuprofen in the waste water of a hospital in Hanoi.     

Liquid–liquid extraction of copper(II) with substituted salicylideneanilines from sulfate media

The liquid–liquid extraction of copper(II) with Schiff bases in chloroform from sulfate media is studied for pH and concentration of the extractant. Stoichiometry coefficients of the extracted species are determined by the slope analysis method. With salicylideneaniline, the copper(II) is extracted as a mixed chelate complex, CuL₂HL. In the presence of substituent, the copper is extracted as simple chelates, CuL₂. The trends in the values of extraction constants were explained in terms of the nature of the substituents.     

Untersuchungen zum Verhalten der Pt-Metalle gegenüber festen und flüssigen Kationenaustauschern mit Hilfe der Radiotracermethode

Mit Hilfe radioaktiver Nuklide wurde die Aufnahme der Pt-Metalle aus chloridhaltigen Lösungen sowoht durch feste (Wofatit KPS) als auch durch flüssige Kationenaustauscher (Bis-2-äthylhexylphosphorsäure) bei Variation des pH-Wertes, der Vorbehandlung sowie der Edelmetallkonzentration bestimmt. Pt und Ir wurden nur geringfügig, Pd und Rh dagegen deutlich stärker adsorbiert. Beide Austauschertypen zeigten qualitativ das gleiche Verhalten. Die gefundenen Abhängigkeiten der Adsorption wurden auf das komplexchemische Verhalten der Pt-Metalle zurückgeführt. Es werden Verfahren zur schnellen radiochemischen Reinigung der benutzten Nuklide angegeben.