Hyphenation of capillary high-performance liquid chromatography with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for nano-scale screening of single-bead combinatorial libraries

This paper focuses on the technical aspects of chemical screening from 384-well plate nano-scale single-bead combinatorial libraries. The analytical technique utilized is a combination of capillary liquid chromatography with ultraviolet detection and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. The HPLC/MALDI-MS hyphenation is achieved by means of a micro-fraction collector with a peak detection system that automatically collects the peaks onto the MALDI targets for subsequent characterization. Several experimental parameters such as type of 384-well plate, well-plate sealing foils, and a column-switching procedure were investigated using a small test library of nine components. Additionally, the influence of different MALDI matrices, different MALDI targets and sample-spotting techniques on the MALDI detection sensitivity as well as the ruggedness and sample throughput capacity of this technique were studied. Optimum results for the analytes investigated were obtained with 2,5-dihydroxybenzoic acid using on-line mixing of HPLC effluent and matrix solution. To demonstrate the potential of this capillary HPLC/MALDI-TOFMS method, its application to several single-bead libraries was investigated. The instrumental method allowed for the rapid identification and purity assessment of combinatorial libraries with detection limits down to the higher femtomole level using both UV detection and MALDI mass spectrometry.     

Rb2Co3(H2O)2[B4P6O24(OH)2]: Ein Borophosphat mit ‐Tetraeder‐Anionenteilstruktur und Oktaeder‐Trimeren (CoO12(H2O)2)

Rb₂Co₃(H₂O)₂[B₄P₆O₂₄(OH)₂]: A Borophosphate with ‐Tetrahedral Anionic Partial Structure and Trimers of Octahedra (Co O₁₂(H₂O)₂) Rb₂Co₃(H₂O)₂[B₄P₆O₂₄(OH)₂] is formed under mild hydrothermal conditions (T = 165 °C) from mixtures of RbOH_(aq), CoCl₂, H₃BO₃, and H₃PO₄ (molar ratio 1 : 1 : 1 : 2). The crystal structure (orthorhombic system) was solved by X‐ray single crystal methods (space group Pbca, No. 61; R‐values (all data): R1 = 0.0699, wR2 = 0.0878): a = 950.1(1) pm, b = 1227.2(2) pm, c = 2007.4(2) pm; Z = 4. The anionic partial structure consists of tetrahedral [B₄P₆O₂₄(OH)₂^8–] layers, which contain three‐ and nine‐membered rings. Co^II is octahedrally coordinated by oxygen and oxygen and H₂O ligands, respectively (coordination octahedra CoO₆ and CoO₄(H₂O)₂). Three adjacent coordination octahedra are condensed via common edges to form trimeric units (CoO₁₂(H₂O)₂). The oxidation state +2 of cobalt was confirmed by magnetic measurements. The octahedral trimers are quasi‐isolated. No long‐range magnetic ordering occurs down to 2 K. Rb⁺ is disordered over three crystallographically independent sites within channels of the structure running parallel [010]; the coordination sphere of Rb⁺ is formed by nine oxygen species of the tetrahedral layers, one OH group and one H₂O molecule.     

Electron-transfer reduction of selected alcohols with alkalide K, K(15-crown-5)2 via organometallic intermediates

The course of the reaction of alkalide K⁻, K⁺(15-crown-5)₂1 with selected alcohols depends on the kind of alcohol and the mode of substrate delivery. In the case of methanol, potassium methoxide formed initially undergoes destruction at the excess of 1. It results in potassium oxide and methylpotassium. The latter opens the crown ether ring giving potassium tetraethylene glycoxide vinyl ether and methane. A similar course of the process is observed for propanol. Potassium glycidoxide is the main product formed in the reaction of 1 with glycidol. Its oxirane ring is opened at the excess of 1. Organopotassium alkoxides, i.e., potassium potassiomethoxide and dipotassium potassiopropane-1,2-dioxide are intermediate products of this reaction. They react then with the crown ether. Potassium methoxide, potassium enolate of acetaldehyde, dipotassium propane-1,2-dioxide and potassium tetraethylene glycoxide vinyl ether are the final products of this process.     

Insights on the origin of the structural phase transition in BaV10O15 from electronic structure calculations and the effect of Ti-doping on its structure and electrical transport properties

Band structure calculations at the level of LMTO-ASA provide insight into the electronic structure of BaV₁₀O₁₅ and the origin of the structural phase transition. A crystal orbital Hamiltonian population/integrated crystal orbital Hamiltonian population analysis provides evidence that the crystallographic phase transition is driven by V-V bond formation. As well, the energy bands near the Fermi level are very narrow, <1 eV, consistent with the fact that the observed insulating behavior can be due to electron localization via either Mott-Hubbard correlation and/or Anderson disorder. The partial solid solution, BaV_10−xTi_xO₁₅, was examined to study the effect of Ti-doping at the V sites on the structure and electronic transport properties. In spite of the non-existence of “BaTi₁₀O₁₅”, the limiting x=8, as indicated by a monotonic increase in the cell volume and systematic changes in properties. This limit may be due to the difficulty of stabilizing Ti²⁺ in this structure. For x=0.5 both the first order structural phase transition and the magnetic transition at 40 K are quenched. The samples obey the Curie-Weiss law to x=3 with nearly spin only effective moments along with θ values which range from −1090 K (x=0.5) to −1629 K (x=3). For x>3 a very large, ∼2×10⁻³ emu/mol, temperature independent (TIP) contribution dominates. Conductivity measurements on sintered, polycrystalline samples show semiconducting behavior for all compositions. Activation energies for Mott hopping derived from high temperature data range from ∼0.1 eV for x=0-1 and fall to a plateau of 0.06 eV for x=3-7. Low temperature data for x=3, 5 and 7 show evidence for Mott variable range hoping (VRH) with a T^1/4 law and in one case between 5 and 17 K, a Efros-Shklovskii correlated hopping, T^1/2 law, was seen, in sharp contrast to BaV₁₀O₁₅ where only the E-S law was observed up to 75 K. Seebeck coefficients are small (<35 μV/K), positive, roughly TIP and increase with increasing x up to x=5. This may point to a Heikes hopping of holes but a simple single carrier model is impossible. The compositions for x>3 are remarkable in that local moment behavior is lost, yet a metallic state is not reached. The failure of this system to be driven metallic even at such high doping levels is not fully understood but it seems clear that disorder induced carrier localization plays a major role.     

Vesicle adsorption and lipid bilayer formation on glass studied by atomic force microscopy

The adsorption of phosphatidylcholine (PC) vesicles (30, 50, and 100 nm nominal diameters) and of dye-labeled PC vesicles (labeled with 6% Texas Red fluorophore (TR) and encapsulated carboxy fluorescein (CF)) to glass surfaces was studied by contact mode atomic force microscopy in aqueous buffer. These studies were performed in part to unravel details of the previously observed isolated rupture of dye-labeled PC vesicles on glass (Johnson, J. M.; Ha, T.; Chu, S.; Boxer, S. G. Biophys. J. 2002, 83, 3371-3379), specifically to differentiate partial rupture, that is, pore formation and leakage of entrapped dye, from full rupture to form bilayer disks. In addition, the adhesion potential of PC vesicles on glass was calculated based upon the adhesion-driven flattening of adsorbed vesicles and a newly developed theoretical model. The vesicles were found to flatten considerably upon adsorption to glass (width-to-height ratio of approximately 5), which leads to an estimate for the adhesion potential and for the critical rupture radius of 1.5 x 10(-4) J/m2 and 250 nm, respectively. Independent of vesicle size and loading with dye molecules, the adsorption of intact vesicles was observed at all concentrations below a threshold concentration, above which the formation of smooth lipid bilayers occurred. In conjunction with previous work (Johnson, J. M.; Ha, T.; Chu, S.; Boxer, S. G. Biophys. J. 2002, 83, 3371-3379), these data show that 6% TR 20 mM CF vesicles adsorb to the surface intact but undergo partial rupture in which they exchange content with the external buffer.     

Bioactivation of self-immolative dendritic prodrugs by catalytic antibody 38C2

Self-immolative dendrimers have recently been developed and introduced as a potential platform for a multi-prodrug. These unique structural dendrimers can release all of their tail units, through a self-immolative chain fragmentation, which is initiated by a single cleavage at the dendrimer's core. Incorporation of drug molecules as the tail units and an enzyme substrate as the trigger can generate a multi-prodrug unit that will be activated with a single enzymatic cleavage. We have synthesized the first generation of dendritic prodrugs with doxorubicin and camptothecin as tail units and a retro-aldol retro-Michael focal trigger, which can be cleaved by catalytic antibody 38C2. The bioactivation of the dendritic prodrugs was evaluated in cell-growth inhibition assay with the Molt-3 leukemia cell line in the presence and the absence of antibody 38C2. The dendritic unit was applied as a platform for a heterodimeric prodrug, which achieved a remarkable increase in toxicity with its bioactivation.     

LiAlH4: From Stoichiometric Reduction to Imine Hydrogenation Catalysis

Imine‐to‐amine conversion with catalytic instead of stoichiometric quantities of LiAlH₄ is demonstrated (85 °C, catalyst loading≥2.5 mol %, pressure≥1 bar). The effects of temperature, pressure, solvent, and catalyst modifications, as well as the substrate scope are discussed. Experimental investigations and preliminary DFT calculations suggest that the catalytically active species is generated in situ: LiAlH₄+Ph(H)C=NtBu→LiAlH₂[N(tBu)CH₂Ph]₂. A cooperative mechanism in which Li and Al both play a prominent role is proposed.     

Fluid phase equilibria of binary and ternary mixtures of supercritical carbon dioxide with tetradecanoic acid and docosane up to 43 MPa and 393 K: cosolvency effect and miscibility windows

Isothermal pressure (p)-mass fraction (w) phase diagrams were measured for CO₂ + tetradecanoic acid at six temperatures from 328.2 K to 373.2 K and for CO₂ + docosane at four temperatures from 343.2 K to 393.2 K as well as isobaric temperature (T)-mass fraction (w) phase diagrams for both systems at 34.5 MPa. In addition the isothermal and isobaric Gibbs phase prisms at 373.2 K and 34.5 MPa respectively were determined for the ternary system CO₂ + tetradecanoic acid + docosane, and and isobaric miscibility window was found between 333 K and 385 K at 34.5 MPa.