Assay of free captopril in human plasma as monobromobimane derivative,using RPLC/(+)ESI/MS/MS: validation aspects and bioequivalence evaluation

A sensitive method for determination of free captopril as monobromobimane derivative in plasma samples is discussed. The internal standard (IS) was 5‐methoxy‐1H‐benzimidazole‐2‐thiol. Derivatization with monobromobimane immediately after blood collection and plasma preparation prevents oxidation of captopril to the corresponding disulfide compound and enhances the ionization yield. Consequently, derivatization enhances sample stability and detection sensitivity. Addition of the internal standard was made immediately after plasma preparation. The internal standard was also derivatized by monobromobimane, as it contains a thiol functional group. Preparation of plasma samples containing captopril and IS derivatives was based upon protein precipitation through addition of acetonitrile, in a volumetric ratio 1:2. The reversed‐phase liquid chromatographic separation was achieved on a rapid resolution cartridge Zorbax SB‐C₁₈, monitored through positive electrospray ionization and tandem MS detection using the multiple‐reaction monitoring mode. Transitions were 408–362 amu for the captopril derivative and 371–260 amu for the internal standard derivative. The kinetics of captopril oxidation to the corresponding disulfide compound in plasma matrix was also studied using the proposed method. A linear log–log calibration was obtained over the concentration interval 2.5–750 ng/mL. A low limit of quantitation in the 2.5 ng/mL range was obtained. The analytical method was fully validated and successfully applied in a three‐way, three‐period, single‐dose (50 mg), block‐randomized bioequivalence study for two pharmaceutical formulations (captopril LPH 25 and 50 mg) against the comparator Capoten 50 mg. Copyright © 2009 John Wiley & Sons, Ltd.     

Reversible transformation of ZnII coordination geometry in a single crystal of porous metal-organic framework [Zn3(ntb)2(EtOH)2].4 EtOH

A 3D porous metal-organic framework [Zn3(ntb)2(EtOH)2]n.4nEtOH (1) that generates 1D channels of honeycomb aperture has been prepared by the solvothermal reaction of Zn(NO3)(2).6 H2O and 4,4',4'-nitrilotrisbenzoic acid (H3NTB) in EtOH at 110 degrees C. Framework 1 exhibits reversible single-crystal-to-single-crystal transformations upon removal and rebinding of the coordinating EtOH as well as the EtOH guest molecules, which give rise to desolvated crystal [Zn3(ntb)2]n (1') and resolvated crystal [Zn3(ntb)2-(EtOH)2]n.4nEtOH (1'). The X-ray structures indicate that 3D host framework is retained during the transformations from 1 to 1' and from 1' to 1', but the coordination geometry of ZnII ions changes from/to trigonal bipyramid to/from tetrahedron, concomitant with the rotational rearrangement of a carboxylate plane of the NTB3- relative to its associated phenyl ring. To retain the single crystal integrity, extensive cooperative motions must exist between the molecular components throughout the crystal. Framework 1' exhibits permanent porosity, thermal stability up to 400 degrees C, and blue luminescence, and high storage capabilities for N2, H2, CO2, and CH4.     

Detached eddy simulation of flow around two wall-mounted cubes in tandem

We investigate the performance of unsteady Reynolds-averaged Navier–Stokes (URANS) computation and various versions of detached eddy simulation (DES) in resolving coherent structures in turbulent flow around two cubes mounted in tandem on a flat plate at Reynolds number (Re) of 22,000 and for a thin incoming boundary layer. Calculations are carried out using four different coherent structure resolving turbulence models: (1) URANS with the Spalart–Allmaras model; (2) the standard DES [Spalart, P.R., Jou, W.H., Strelets, M., Allmaras, S.R., 1997. Comments on the feasibility of LES for wings, and on a hybrid RANS/LES approach. In: Liu, C., Liu, Z., (Eds.), Advances in DNS/LES. Greyden Press, Columbus, OH]; (3) the Delayed DES (DDES); and (4) the DES with a low-Re modification (DES-LR) [Spalart, P., Deck, S., Shur, M., Squires, K., Strelets, M., Travin, A., 2006. A new version of detached eddy simulation, resistant to ambiguous grid densities. Theor. Comput. Fluid Dyn. 20 (3), 181–195]. The grid sensitivity of the computed solutions is examined by carrying out simulations on two successively refined grids. The computed results for all cases are compared with the experimental measurements of Martinuzzi and Havel [Martinuzzi, R., Havel, B., 2000. Turbulent flow around two interfering surface-mounted cubic obstacles in tandem arrangement. ASME J. Fluids Eng. 122, 24–31] for two different cube spacings. All turbulence models reproduce essentially identical separation of the approach thin boundary layer and yield an unsteady horseshoe vortex system consisting of multiple vortices in the leading edge region of the upstream cube. Significant discrepancies between the URANS and all DES solutions are observed, however, in other regions of interest such as the shear layers emanating from the cubes, the inter-cube gap and the downstream wake. Regardless of the grid refinement, URANS fails to capture key features of the mean flow, including the second horseshoe vortex in the upstream junction and recirculating flow on the top surface of the downstream cube for the large cube spacing, and underestimates significantly turbulence statistics in most regions of the flow for both cases. On the coarse mesh, all three DES approaches appear to yield very similar results and fail to reproduce the second horseshoe vortex. The standard DES and DDES solutions obtained on the fine meshes are essentially identical and both suffer from premature switching to unresolved DNS, due to the mis-interpretation of grid refinement as wall proximity, which leads to spurious vortices in the inter-cube region. Numerical solutions show that the low-Re modification (DES-LR) is critical prerequisite in DES on the ambiguously fine – not fine enough for full LES – mesh to prevent excessive nonlinear drop of the subgrid eddy viscosity in low cell-Re regions like in the inter-obstacle gap. Mean flow quantities and turbulence statistics obtained with DES-LR on the fine mesh are in good overall agreement with the measurements in most regions of interest for both cases.     

Barium Sodium Niobate Powders and Thin Films: Preparation and Ferroelectric Behavior

Powders and thin films of barium sodium niobate, Ba₄Na₂Nb₁₀O₃₀, of filled tungsten bronze type ferroelectric were processed by a sol-gel route using barium metal, niobium ethoxide and sodium methoxide as precursors. Polycrystalline powder resulted after heat treating the gel powder at or above 650°C. Thin films of Ba₄Na₂Nb₁₀O₃₀ showed no preferred orientation on Si(100), Pt/Si(100) and sapphire substrates. Hysteresis measurements at 1 kHz for the thin films annealed at 750°C, obtained using a prehydrolyzed precursor solution, and gave remanent polarization of 17.34 µC/cm² and coercive field of 62.5 kV/cm. Microstructural investigation of surface morphology of these films revealed grains about 0.3 µm in size. Prehydrolysis of the precursor solution was found to be necessary to achieve dense films with ferroelectric properties.     

Validation of the Metabolite Ergothioneine as a Forensic Marker in Bloodstains

Ergothioneine, which is a naturally occurring metabolite, generally accumulates in tissues and cells subjected to oxidative stress, owing to its structural stability at physiological pH; therefore, it has been attracting attention in various biomedical fields. Ergothioneine has also been suggested as a potential forensic marker, but its applicability has not yet been quantitatively validated. In this study, quantitative analysis of ergothioneine in bloodstains was conducted to estimate the age of bloodstains and that of bloodstain donors. Blood from youth and elderly participants was used to generate bloodstains. After extracting metabolites from the bloodstains under prevalent age conditions, ergothioneine levels were quantified by mass spectrometry via multiple reaction monitoring. The concentration of ergothioneine in day 0 bloodstains (fresh blood), was significantly higher in the elderly group than in the youth group, but it did not differ by sex. Statistically significant differences were observed between the samples from the two age groups on days 0, 5 and 7, and on days 2 and 3 compared with day 0. The findings suggest that ergothioneine can be used to estimate the age of bloodstains and of the donor; it could be useful as a potential marker in reconstructing crime scenes.     

An Fc variant with two mutations confers prolonged serum half-life and enhanced effector functions on IgG antibodies

The pH-selective interaction between the immunoglobulin G (IgG) fragment crystallizable region (Fc region) and the neonatal Fc receptor (FcRn) is critical for prolonging the circulating half-lives of IgG molecules through intracellular trafficking and recycling. By using directed evolution, we successfully identified Fc mutations that improve the pH-dependent binding of human FcRn and prolong the serum persistence of a model IgG antibody and an Fc-fusion protein. Strikingly, trastuzumab-PFc29 and aflibercept-PFc29, a model therapeutic IgG antibody and an Fc-fusion protein, respectively, when combined with our engineered Fc (Q311R/M428L), both exhibited significantly higher serum half-lives in human FcRn transgenic mice than their counterparts with wild-type Fc. Moreover, in a cynomolgus monkey model, trastuzumab-PFc29 displayed a superior pharmacokinetic profile to that of both trastuzumab-YTE and trastuzumab-LS, which contain the well-validated serum half-life extension Fcs YTE (M252Y/S254T/T256E) and LS (M428L/N434S), respectively. Furthermore, the introduction of two identified mutations of PFc29 (Q311R/M428L) into the model antibodies enhanced both complement-dependent cytotoxicity and antibody-dependent cell-mediated cytotoxicity activity, which are triggered by the association between IgG Fc and Fc binding ligands and are critical for clearing cancer cells. In addition, the effector functions could be turned off by combining the two mutations of PFc29 with effector function-silencing mutations, but the antibodies maintained their excellent pH-dependent human FcRn binding profile. We expect our Fc variants to be an excellent tool for enhancing the pharmacokinetic profiles and potencies of various therapeutic antibodies and Fc-fusion proteins.Subject terms: Antibody therapy, Molecularly targeted therapy, Drug development     

On Rescaled Poisson Processes and the Brownian Bridge

The process obtained by rescaling a homogeneous Poisson process by the maximum likelihood estimate of its intensity is shown to have surprisingly strong self-correcting behavior. Formulas for the conditional intensity and moments of the rescaled Poisson process are derived, and its behavior is demonstrated using simulations. Relationships to the Brownian bridge are explored, and implications for point process residual analysis are discussed.     

On Non-simple Marked Point Processes

Simple point processes are often characterized by their associated compensators or conditional intensities. Non-simple point processes are not uniquely determined by their conditional intensity and compensator, so instead one may identify with the point process its associated simple point process and corresponding conditional intensity, on an expanded mark space. Some relations between the conditional intensity on the expanded mark space and the ordinary conditional intensity are investigated here, and some classes of separable non-simple processes are presented. Transformations into simple point processes, involving thinning and rescaling, are presented.     

Reaction of diphenylcyclopropenethione with Mn(CO)5−: Ring cleavage,alkylation, and reduction

Alkylation, reduction, and ring opening of diphenylcyclopropenethione to a π-allyl complex occurs on treatment with Mn(CO)₅^? and methyl iodide in aqueous methanol.