μLC coupled to ICP–SFMS with post-column isotope dilution analysis of sulfur for absolute protein quantification

Absolute protein quantification has become an important challenge in modern bioanalytical chemistry. Among several approaches based on mass spectrometric techniques, inductively coupled plasma (ICP) as ionisation source provides element-selective and sensitive detection of heteroatoms, and thus, a potentially emerging tool in protein analysis. In this work we applied coupling of capillary liquid chromatography (μLC) and inductively coupled plasma-sector field mass spectrometry (ICP–SFMS) to the separation and determination of standard proteins. For quantification purposes, post-column isotope dilution of sulfur was applied and optimised for this type of hyphenated technique. Provided that the protein sequence is known (number of sulfur-containing amino acids, i.e. cysteines and methionines) the protein amount can then be directly calculated from the determined sulfur content in a certain protein fraction. In order to prove the reliability of the presented method, two different certified reference materials were analysed: CRM 393 (human apolipoprotein A-I) and CRM 486 (α-fetoprotein). For CRM 393 excellent agreement (37.0 ± 1.4 μmol L⁻¹) was obtained with the certificate (37.7 ± 1.8 μmol L⁻¹). However, the recovery rate for α-fetoprotein in CRM 486 was found to be about 60% indicating incomplete elution of the protein during the chromatographic separation.     

In search of novel ligands using a structure-based approach: a case study on the adenosine A<Subscript>2A</Subscript> receptor

In this work, we present a case study to explore the challenges associated with finding novel molecules for a receptor that has been studied in depth and has a wealth of chemical information available. Specifically, we apply a previously described protocol that incorporates explicit water molecules in the ligand binding site to prospectively screen over 2.5 million drug-like and lead-like compounds from the commercially available eMolecules database in search of novel binders to the adenosine A_2A receptor (A_2AAR). A total of seventy-one compounds were selected for purchase and biochemical assaying based on high ligand efficiency and high novelty (Tanimoto coefficient ≤0.25 to any A_2AAR tested compound). These molecules were then tested for their affinity to the adenosine A_2A receptor in a radioligand binding assay. We identified two hits that fulfilled the criterion of ~50 % radioligand displacement at a concentration of 10 μM. Next we selected an additional eight novel molecules that were predicted to make a bidentate interaction with Asn253^6.55, a key interacting residue in the binding pocket of the A_2AAR. None of these eight molecules were found to be active. Based on these results we discuss the advantages of structure-based methods and the challenges associated with finding chemically novel molecules for well-explored targets.     

On computing dominant frequency from bipolar intracardiac electrograms

Dominant frequency (DF) computed from action potentials is a key parameter for investigating atrial fibrillation in animal studies and computer models. A recent clinical trial reported consistent results computing DF from 30 Hz to 400 Hz bandpass filtered bipolar electrograms in humans. The DF (< 15 Hz and, thus, filtered out) was recovered by rectifying the signal, while the theoretical background of this approach was left uncommented. It is the focus of this paper to provide this background by a Fourier analysis. We demonstrate that it is mainly the timing of the narrow deflections (local activation at the catheter tip) which contribute to the DF peak in the frequency spectrum. Due to the typical signal morphology pronounced harmonic peaks occur in the spectrum. This is a disadvantage when computing the regularity index (RI) as a parameter for local organization and signal quality. It is demonstrated for synthetical and patient data that at low DF the RI is far below the optimal value one even for high underlying organization and good signal quality. The insight obtained promotes the development of better measures for organization. The finding that mainly timing of activation contributes to DF might promote the development of powerful realtime signal processing tools for computing DF.     

100 Jahre Einkristallzucht aus der Schmelze

Cars, television, mobile phones, digital cameras, cash machines: Daily life is strongly affected by microchips produced from high purity silicon single crystals via thin wafers. Most of these single crystals are prepared by a process invented by the German‐Polish scientist Jan Czochralski in 1916 in the “Kabelwerk Oberspree (KWO)” of the “Allgemeine Elektricitätsgesellschaft (AEG)” in Berlin‐Oberschöneweide. Czochralski discovered the famous method to pull single crystals by accident: Deep in thought, he dipped his pen not into an ink pot but into a crucible with liquid tin, both standing next to one another on his desk. Quickly he pulled his pen out and observed a thin thread of tin emerging from the tip. After etching, the thread was identified as a single crystal of tin. This observation is probably one of the most important technical inventions of the first half of the 20^th century. In 1917, he left the AEG in Berlin and worked in the metal research laboratory, later belonging to the “Metallgesellschaft”, in Frankfurt/Main. Until today, wafers of high‐purity silicon are prepared by the Czochralski method. Silicon wafers with 200 mm diameter were produced in 1990, 300 mm wafers in 2001. The production of wafers with 450 mm diameter was expected for 2016. Siltronic produced in 2009 the first dislocation‐free silicon single crystal with 450 mm diameter, and other companies followed. However, until now, the 450 mm technology is not standard. This is due to a combination of very high investment costs needed to establish the 450 mm technology and very low prices of microchips.     

A Benchtop Method for Appending Protic Functional Groups to N-Heterocyclic Carbene Protected Gold Nanoparticles

The remarkable resilience of N-heterocyclic carbene (NHC) gold bonds has quickly made NHCs the ligand of choice when functionalizing gold surfaces. Despite rapid progress using deposition from free or CO₂-protected NHCs, synthetic challenges hinder the functionalization of NHC surfaces with protic functional groups, such as alcohols and amines, particularly on larger nanoparticles. Here, we synthesize NHC-functionalized gold surfaces from gold(I) NHC complexes and aqueous nanoparticles without the need for additional reagents, enabling otherwise difficult functional groups to be appended to the carbene. The resilience of the NHC−Au bond allows for multi-step post-synthetic modification. Beginning with the nitro-NHC, we form an amine-NHC terminated surface, which further undergoes amide coupling with carboxylic acids. The simplicity of this approach, its compatibility with aqueous nanoparticle solutions, and its ability to yield protic functionality, greatly expands the potential of NHC-functionalized noble metal surfaces.     

Nano-liquid chromatography-direct electron ionization mass spectrometry: improving performance by a new ion source adapter

A novel interface adapter has been designed to provide a new way of directly coupling a nano-liquid chromatograph to an electron ionization mass spectrometer. It connects the transfer capillary coming from the liquid chromatograph to the ionization chamber and can be easily screwed into the ion source. Liquid coming from the column passes through the heated adapter flow path and is vaporized. A continuous flow of new liquid pushes the vapor into the ionization chamber where it is ionized and continues on to the mass analyzer. The advantages of the new adapter are reduced ice formation inside the ion source and less clogging of the transfer capillary. Improvements achieved are demonstrated on the basis of caffeine and steroid analysis. The limits of detection of selected steroids are compared with and without the adapter. The adapter improves the detection limit of the system by a factor of 2 and precision from ≤15% to ≤9% relative standard deviation. No derivatization procedure is necessary before the analysis of small polar compounds. The resulting spectra are reproducible, easily interpretable, and database searchable. The new method is robust, delivers reproducible results, and provides a highly efficient alternative to existing methods in the field of pharmaceutical analysis.     

Der Wittelsbacher und der Hope‐Diamant

Diamonds are formed from carbon at high pressures and high temperatures in the inner part of the earth. Doping with very small amounts of boron leads to diamonds with blue colour. Two of the most famous historical blue diamonds, the Wittelsbach and Hope Diamond, were found in the Indian Kollur mine. The latter was brought to Europe by the French gem merchant Tavernier. Today it is displayed in the Smithsonian Institute. The Wittelsbach Diamond was for a long time in the possession of the House Wittelsbach until it was secretly sold in Antwerp in 1951. In 2008, it was purchased by auction by the jeweller Graff who recut the gem. In 2011, it was sold to an unknown buyer. As the Wittelsbach and the Hope diamond share origin and colour, it was assumed for a long time that both are pieces from a larger crystal. By optical investigation it was now shown that they have indeed some similar optical properties, but differ strikingly in other ones. Hence, they cannot originate from the same crystal.