An affinity-based probe for the proteomic profiling of aspartic proteases

We have developed an affinity-based probe for the proteomic profiling of aspartic proteases. Our probe was shown to be selective towards aspartic proteases over other proteins. It was also shown that the strategy may be used to label selectively aspartic proteases in the presence of a large excess of other proteins, thus making it useful for future proteome profiling experiments.     

XPS in‐depth composition study on commercial monocrystalline silicon solar cells

From large‐scale production, two monocrystalline silicon solar cells of different quality, i.e. I_SC = 3.0 A (good cell) and I_SC = 1.6 A (bad cell), have been studied by XPS combined with 4 keV Ar⁺ depth profiling. Depth profiling was carried out through the anti‐reflection coating (TiO₂), the passivation layer (SiO₂) and up into the phosphorus‐doped silicon bulk. At the solar cell surface the elemental composition is similar for both cells, although the bad one presents slightly more carbon, phosphorus and lead but less silver than the good one. During profiling, carbon and silver could be followed by XPS. It was found that the carbon content is distinguishably higher in the bad cell than in the good one. Furthermore, it was found that silver atoms have not diffused in the same way in both cells. Only the good cell presents silver atoms up into the silicon bulk. Copyright © 2003 John Wiley & Sons, Ltd.     

Depth profiles of shallow implanted layers by soft ion sputtering and total-reflection X-ray fluorescence

A new method suitable for depth profiling of shallow layers on different materials is presented. It is based on a soft and planar ion sputtering combined with differential weighing, total-reflection X-ray fluorescence (TXRF) spectrometry and Tolansky interferometry. By means of a stepwise repetition of these techniques it is possible to determine both density/depth and concentration/depth profiles. The respective quantities are expressed in terms inherent only to the sample and traceable to the SI-units or subunits gram, nanometer and mole. It is a unique feature of this method that density/depth profiles can directly be obtained from measurements without any calibration or theoretical approximation. The method is applied to a Si wafer implanted with Co ions of 25 keV energy and a nominal dose of 1×10¹⁶ cm⁻². The depth resolution is shown to be <3 nm while a total depth of some 100 nm can be reached. The concentration/depth profile is compared with RBS measurements, wet-chemical etching plus TXRF and Monte Carlo simulations. In view of the fact that only similar but not exactly the same samples have been examined by these methods, a good correspondence can be noticed.     

Chemical and analytical characterization of related organic impurities in drugs

A system is proposed for the classification of related organic impurities in drugs and drug products including among others (separated and non-separated) intermediates, various kinds of by-products, among them products of different side reactions, epimeric/diastereomeric, enantiomeric impurities, impurities in natural products, and finally degradation products. Examples are taken mainly from the author's own experience and from among the named impurities in the European Pharmacopoeia with focus on impurities in hydrocortisone, prednisolone, enalapril maleate, lisinopril, ethynodiol diacetate, pipecuronium bromide, cimetidine, and ethynylsteroids. The methodological aspects of impurity profiling from the detection to the identification/structure elucidation and quantitative determination of impurities are briefly summarized.This paper is Part 23 in the series "Estimation of impurity profiles of drugs and related materials". For Part 22 see ref. [1].     

Cathodoluminescence Depth Profiling in SiO2:Ge Layers

 For investigation of the luminescent center profile cathodoluminescence measurements are used under variation of the primary electron energy E ₀ = 2…30 keV. Applying a constant incident power regime (E ₀·I ₀ = const), the depth profiles of luminescent centers are deduced from the range of the electron energy transfer profiles dE/dx. Thermally grown SiO₂ layers of thickness d = 500 nm have been implanted by Ge⁺-ions of energy 350 keV and doses (0.5–5)10¹⁶ ions/cm². Thus Ge profiles with a concentration maximum of (0.4 – 4) at% at the depth of d_m≅240 nm are expected. Afterwards the layers have been partially annealed up to T _a = 1100 °C for one hour in dry nitrogen. After thermal annealing, not only the typical violet luminescence (λ = 400 nm) of the Ge centers is strongly increased but also the luminescent center profiles are shifted from about 250 nm to 170 nm depth towards the surface. This process should be described by Ge diffusion processes, precipitation and finally Ge nanocluster formation. Additionally, a Ge surface layer is piled-up extending to a depth of roughly 25 nm.     

Investigation of Al diffusion in different oxide thin-film layers by SNMS/HFM method

Depth profiles of Ga₂O₃/a-SiO₂/Al₂O₃- substrate, Ga₂O₃/a-Si₃N₄/Al₂O₃- substrate, and Ga₂O₃/Al₂O₃ substrate thin layers were determined by the SNMS/HFM method. Al diffusion from the Al₂O₃ substrate was investigated after 50, and in some cases after 600 hours of heat treatment time at different temperatures (600 °C,850 °C,950 °C,1050 °C and 1150 °C). The diffusion coefficient of Al at 850 °C was found to be D _Al=8.7 * 10^–18 cm²/s in amorphous SiO₂; D _Al=1.5*10^–17 cm²/s in amorphous Si₃N₄ and D _Al=5.5* 10^–16 cm²/s in Ga₂O₃ at 600 °C, respectively. The possible diffusion mechanism is explained in terms of the metal-oxygen bond-strengths. Although the studied materials have high resistivity at room temperature, the applied SNMS/HFM method has proven to be an efficient surface analytical tool even in these cases.Dedicated to Professor Dr. rer. nat. Dr. h.c. Hubertus Nickel on the occasion of his 65th birthday     

A generic approach to the impurity profiling of drugs using standardised and independent capillary zone electrophoresis methods coupled to electrospray ionisation mass spectrometry

Three standardised, capillary zone electrophoresis-electrospray ionisation mass spectrometry (CZE-ESI-MS) methods were developed for the analysis of six drug candidates and their respective process-related impurities comprising a total of 22 analytes with a range of functional groups and lipophilicities. The selected background electrolyte conditions were found to be: 60/40 v/v 10 mM ammonium formate pH 3.5/organic, 60/40 v/v 10 mM ammonium acetate pH 7.0/organic and 10 mM piperidine, pH 10.5, where the organic solvent is 50/50 v/v methanol/acetonitrile. The coaxial sheath flow consisted of either 0.1% v/v formic acid in 50/50 v/v methanol/water, or 10 mM ammonium acetate in 50/50 v/v methanol/water, depending on the mixture being analysed. Factor analysis and informational theory were used to quantify the orthogonality of the methods and predict their complementarities. The three selected CZE-ESI-MS methods allowed the identification of 21 out of 22 of all the drug candidates and their process-related impurities and provided orthogonality with four established high-performance liquid chromatography-mass spectrometry (HPLC-MS) methods. These methodologies therefore form the basis of a generic approach to impurity profiling of pharmaceutical drug candidates and can be applied with little or no analytical method development, thereby offering significant resource and time savings.     

Detection and recognition of cancer cells in vivo

A fundamentally new recognition method of bio-objects (e.g., cancer cells as the most important case of them) that escape the immune system supervision control is suggested. It is proposed to use a unified complex consisting of several molecular groups (e.g., antibodies or their fragments) bound with each other. Binding targets are localized on the surface of this bio-object. The choice of the targets is determined by antigen profiling being expressed on the surface of these bio-objects. The recognition efficiency appears to be notably higher than in a situation when molecular groups do not form a unified complex and act separately.