Profiling primaquine metabolites in primary human hepatocytes using UHPLC‐QTOF‐MS with 13C stable isotope labeling

Therapeutic efficiency and hemolytic toxicity of primaquine (PQ), the only drug available for radical cure of relapsing vivax malaria are believed to be mediated by its metabolites. However, identification of these metabolites has remained a major challenge apparently due to low quantities and their reactive nature. Drug candidates labeled with stable isotopes afford convenient tools for tracking drug‐derived metabolites in complex matrices by liquid chromatography‐tandem mass spectrometry (LC‐MS‐MS) and filtering for masses with twin peaks attributable to the label. This study was undertaken to identify metabolites of PQ from an in vitro incubation of a 1:1 w/w mixture of ¹³C₆‐PQ/PQ with primary human hepatocytes. Acquity ultra‐performance LC (UHPLC) was integrated with QTOF‐MS to combine the efficiency of separation with high sensitivity, selectivity of detection and accurate mass determination. UHPLC retention time, twin mass peaks with difference of 6 (originating from ¹³C₆‐PQ/PQ), and MS‐MS fragmentation pattern were used for phenotyping. Besides carboxy‐PQ (cPQ), formed by oxidative deamination of PQ to an aldehyde and subsequent oxidation, several other metabolites were identified: including PQ alcohol, predictably generated by oxidative deamination of PQ to an aldehyde and subsequent reduction, its acetate and the alcohol's glucuronide conjugate. Trace amounts of quinone‐imine metabolites of PQ and cPQ were also detected which may be generated by hydroxylation of the PQ/cPQ quinoline ring at the 5‐position and subsequent oxidation. These findings shed additional light on the human hepatic metabolism of PQ, and the method can be applied for identification of reactive PQ metabolites generated in vivo in preclinical and clinical studies. Copyright © 2013 John Wiley & Sons, Ltd.     

A theoretical and experimental study of the behaviour of a parametric array in a random medium

The fluctuations of a parametric signal in a random medium have been investigated. A theoretical expression for the mean square amplitude and phase fluctuations has been derived, by using the small perturbation method. A simplified version of the theoretical model, which predicts the axial amplitude fluctuations of the difference frequency signal, was computerized. Laboratory experiments were carried out in a heated tank in which the fluctuations of a parametric signal and a linearly generated signal were compared. A similar experiment was also carried out in an estuary. The experimental results obtained were consistent with the theoretical estimates.     

Time domain investigation of transceiver functions using a known reference target

During August 1998, a bottom scattering tank experiment was performed at the Applied Research Laboratory, University of Texas to measure wideband acoustic reverberation from multiple objects (e.g., cobbles and pebbles) placed on a sediment simulation of the sea floor. In preparation for processing and analyzing the experimental data, time domain scattering measurements made with stainless steel and glass balls suspended in the water column were used to calibrate the sonar transceiver system by deconvolving the theoretical impulse response for steel and glass spheres, obtained via the Faran elastic sphere scattering model, from the scattered time signals. It is the analysis of these calibration measurements which forms the subject of this paper. Results show the critical importance of accurate input-output system calibrations for time domain sound scattering research, and successfully demonstrate a time domain method for accurately calibrating the complete sonar transceiver function, i.e., both the amplitude and time dependence, using a known reference target. The work has implications for boundary and volume scattering applications.     

Acoustic virtual mass of granular media

Mechanical coupling between grains in a randomly packed unconsolidated granular medium is shown to cause an increase in the effective inertia, hence, a reduction in sound and shear wave speeds, relative to predictions by the standard expressions for a uniform elastic solid. The effect may be represented as a virtual mass term, and directly related to the scintillation index of the grain-to-grain contact stiffness.     

Sound amplitude fluctuations due to a temperature microstructure

The experiments reported in this paper were carried out in a water tank in which a random medium was generated by convective mixing from an array of heaters. An approximate thermodynamic model of the medium was derived. Temperature measurements were made which showed that the temperature microstructure created in this way could be considered as a passive additive of turbulence. Furthermore, it was possible to characterize the random refractive index in terms of a spectral distribution by using an adapted version of a spectrum proposed by Medwin for the upper ocean. By using the adapted Medwin model and the single-scatter theoretical results of Tatarski, theoretical estimates were obtained of the fluctuations of an acoustic signal propagating in this particular medium. Experiments were carried out to measure acoustic signal amplitude fluctuations at frequencies of 9 MHz and 1 MHz. The empirical results were in agreement with the theoretical estimates. Measurements are also reported for the spatial correlation functions of the acoustic signal amplitude fluctuations. The results are discussed in the light of currently available theoretical results.     

Finite element modeling of reverberation and transmission loss in shallow water waveguides with rough boundaries

A finite element model for the reverberation and propagation in a shallow water waveguide with a sandy bottom was calculated for five different environments at a center frequency of 250 Hz. The various environments included a rough water/sediment interface, a rough air/water interface, roughness at both interfaces and downward and upward refracting sound speed profiles with roughness at both interfaces. When compared to other models of reverberation such as ray theory, coupled modes, and parabolic equations, finite elements predicted higher levels of reverberation. At early times, this is due to the "fathometer" return, energy that is normally incident on the boundaries at zero range. At later times, the increased reverberation was due to high angle scattering paths between the two interfaces. Differences in reverberation levels among the environments indicated that scattered energy from the air/water interface is transmitted into the bottom at steep angles. This led to a large decrease in reverberation for a rough air/water interface relative to a rough water/sediment interface. Sound speed profile effects on reverberation were minimal at this frequency range. Calculations of the scintillation index of the different environments indicated that most of the reverberation was relatively Rayleigh-like with heavier tailed distributions at longer ranges.     

Shear and compressional wave speeds in Hertzian granular media

It is shown that the shear wave speed in a granular medium is less than that in an elastic solid of the same shear modulus-to-density ratio. Shear and compressional wave speeds are derived for granular media using a conservation of energy approach. The grains are assumed to be spherical with elastic Hertzian contacts of constant stiffness. The affine approximation is used to determine the relative displacements of grain centers, and it is also assumed that the grains are small compared to a wavelength, consistent with the effective medium approximation. Potential and kinetic energies associated with linear motion are the same as those in an elastic solid, but it is found that shear wave propagation in a granular medium involves additional energies associated with grain rotation. The partition of energies results in a reduction in the shear wave speed, relative to an elastic solid of the same shear modulus-to-density ratio. It is shown that the reduction is an inherent property of granular media, independent of any departure from the affine approximation or fluctuations in coordination number or contact stiffness. The predicted wave speed ratios are consistent with published measurements.     

Computer extended series solution for unsteady flow in a contracting or expanding pipe

Unsteady flow in a semi-infinite contracting or expanding pipeis reinvestigated using long series analysis. The proposed seriesmethod is useful in analysing the problem for a moderately largeconstant ( = aa/, where a = a(t), the radius of the pipe isa function of time, a(t) is the velocity of the wall, and iskinematic viscosity). For positive values of (expansion ofthe pipe) accuracy of the series representing shear stress andpressure gradient is increased from = 2.89 to = 6.0 by extractingthe singularity followed by completion of the series. For negativevalues of (contraction of the pipe), we revert the series whichresults into the increase of the region of validity of the transposedseries from = -25.0 to = -2.89. Later we use Padé approximantsfor summing them. Also, the asymptotic solution for large valuesof is obtained and it agrees closely with pure numerical valuesof shear stress at the wall and pressure gradient.