Characterization of inclusion complexes of betamethasone-related steroids with cyclodextrins using high-performance liquid chromatography

HPLC was used to study the inclusion complexes formed between various beta- and gamma-cyclodextrins and a series of corticosteroids related to betamethasone. Apparent association constants were measured in acetonitrile-water for a set of 13 steroids. An increase in the stability of the steroid-cyclodextrin complex is observed at lower concentrations of acetonitrile. The effects of the nature of the halide at the 9-position, the location of a double bond within the C-ring, substitution at the 9- and 11-positions, and modification of the D-ring of the steroid backbone were studied. The 11- and 17-positions were found to be critically involved in the inclusion process. Larger apparent association constants were obtained with gamma-cyclodextrin (gamma-CD) than with beta-cyclodextrin (beta-CD) due to the increased diameter of the gamma-CD cavity. Van't Hoff plots were constructed to examine the thermodynamic properties of the inclusion process. Plots constructed using retention factors were found to be nonlinear when gamma-CD was present in the mobile phase. This is due to an increase in the strength of the inclusion complex as temperature decreases. Plots constructed using apparent association constants were linear, indicating that the mechanism of inclusion does not change over the range of temperatures studied (10 to 80 degrees C). Enthalpy-entropy compensation was observed for 11 of the 13 steroids studied. The usefulness of cyclodextrins to achieve the separation of steroids in HPLC is discussed and a practical application for the analysis of a steroid and three potential impurities is described.     

Pharmacokinetics of ICG and HPPH-car for the detection of normal and tumor tissue using fluorescence, near-infrared reflectance imaging: a case study

We present in vivo fluorescent, near-infrared (NIR), reflectance images of indocyanine green (ICG) and carotene-conjugated 2-devinyl-2-(1-hexyloxyethyl) pyropheophorbide (HPPH-car) to discriminate spontaneous canine adenocarcinoma from normal mammary tissue. Following intravenous administration of 1.0 mg kg-1 ICG or 0.3 mg kg-1 HPPH-car into the canine, a 25 mW, 778 nm or 70 mW, 660 nm laser diode beam, expanded by a diverging lens to approximately 4 cm in diameter, illuminated the surface of the mammary tissue. Successfully propagating to the tissue surface, ICG or HPPH-car fluorescence generated from within the tissue was collected by an image-intensified, charge-coupled device camera fitted with an 830 or 710 nm bandpass interference filter. Upon collecting time-dependent fluorescence images at the tissue surface overlying both normal and diseased tissue volumes, and fitting these images to a pharmacokinetic model describing the uptake (wash-in) and release (wash-out) of fluorescent dye, the pharmacokinetics of fluorescent dye was spatially determined. Mapping the fluorescence intensity owing to ICG indicates that the dye acts as a blood pool or blood persistent agent, for the model parameters show no difference in the ICG uptake rates between normal and diseased tissue regions. The wash-out of ICG was delayed for up to 72 h after intravenous injection in tissue volumes associated with disease, because ICG fluorescence was still detected in the diseased tissue 72 h after injection. In contrast, HPPH-car pharmacokinetics illustrated active uptake into diseased tissues, perhaps owing to the overexpression of LDL receptors associated with the malignant cells. HPPH-car fluorescence was not discernable after 24 h. This work illustrates the ability to monitor the pharmacokinetic delivery of NIR fluorescent dyes within tissue volumes as great as 0.5-1 cm from the tissue surface in order to differentiate normal from diseased tissue volumes on the basis of parameters obtained from the pharmacokinetic models.     

Correlation between the low temperature phonon mean free path and glass transition temperature in amorphous solids

An interpretation is offered for the increased chemical resistance of glass surfaces upon treatment with SiF₄/H₂O gaseous mixtures. Scanning electron microscopy examinations of the morphology and evolution of the surface structure, the surface sodium concentration profile as revealed by a nuclear reaction and ion release experiments support the formation of a very adherent amorphous silica layer - still transparent at the earlier stages of treatment - which fits well all experimental findings.     

Biogenic aldehyde determination by reactive paper spray ionization mass spectrometry

Ionization of aliphatic and aromatic aldehydes is improved by performing simultaneous chemical derivatization using 4-aminophenol to produce charged iminium ions during paper spray ionization. Accelerated reactions occur in the microdroplets generated during the paper spray ionization event for the tested aldehydes (formaldehyde, n-pentanaldehyde, n-nonanaldehyde, n-decanaldehyde, n-dodecanaldehyde, benzaldehyde, m-anisaldehyde, and p-hydroxybenzaldehyde). Tandem mass spectrometric analysis of the iminium ions using collision-induced dissociation demonstrated that straight chain aldehydes give a characteristic fragment at m/z 122 (shown to correspond to protonated 4-(methyleneamino)phenol), while the aromatic aldehyde iminium ions fragment to give a characteristic product ion at m/z 120. These features allow straightforward identification of linear and aromatic aldehydes. Quantitative analysis of n-nonaldehyde using a benchtop mass spectrometer demonstrated a linear response over 3 orders of magnitude from 2.5 ng to 5 μg of aldehyde loaded on the filter paper emitter. The limit of detection was determined to be 2.2 ng for this aldehyde. The method had a precision of 22%, relative standard deviation. The experiment was also implemented using a portable ion trap mass spectrometer.     

Measurement of turbulent wall velocity gradients using cylindrical waves of laser light

A new optical instrument has been developed for direct measurement of instantaneous velocity gradients at the bounding wall. Light emerging from two tiny optical slits in the surface is used to form a fan of fringes in the region very near the wall. Doppler frequency of the light scattered by the seed particles is directly proportional to the velocity gradient. The system has been used to measure the statistics of the streamwise and spanwise velocity gradients in a turbulent boundary layer. The streamwise and spanwise rms fluctuations were found to be 38% and 11% of the mean streamwise value respectively. The latter result is subject to a large uncertainty.List of symbols a slit width - B transfer function of the instrument - B ^* normalized transfer function - path-averaged value of the normalized transfer function - c constant in logarithmic velocity profile - C _f skin friction coefficient - d _f fringe spacing - f ₁,f₂ frequencies at the downstream and upstream slits resp. - f _d heterodyne Doppler frequency of the signal - g(t) instantaneous wall velocity gradient - G Clauser shape factor - mean wall velocity gradient - g rms value of the wall velocity gradient - H boundary layer shape factor - i, j, k unit vectors along x, y, z axes - wavenumber of laser light - L major axis of the elliptic cross-section of the laser sheet at the slit - l length of each slit - N number of cycles in a signal - N ₀ number of cycles without frequency-shifting - n difference of the unit vectors u ₁and u ₂ - P power transmitted through a slit - P _o power incident on a slit - Re ₁ Reynolds number based on displacement thickness and free-stream velocity - Re ₂ Reynolds number based on momentum thickness and free-stream velocity - S spacing between the slits - S ^* normalized spacing between the slits - u streamwise velocity - u ₁,u₂ unit vectors along the local directions of propagation of the two cylindrical waves - u _l linear term in the streamwise velocity profile - u _nl nonlinear terms in the streamwise velocity - u _nl ^* normalized value of nonlinear streamwise velocity - u _nl ^* mean streamwise velocity - u friction velocity - u⁺ mean velocity normalized with friction velocity - v velocity component normal to the wall - v ^* normal velocity normalized with streamwise velocity - V velocity vector - w spanwise component of velocity - W minor axis of the elliptic cross-section of the laser sheet at the slit - x streamwise distance - ± x _m limiting values of streamwise distance for a signal - x ^* normalized streamwise distance - x ^* normalized value of x _m - y normal distance - y ⁺ normal distance normalized with friction length scale - z spanwise distance - z ⁺ spanwise distance normalized with friction length scale - half-spreading angle of the cylindrical waves - boundary layer thickness in Coles' profile - ₁ displacement thickness of the boundary layer - ₂ momentum thickness of the boundary layer - ₃ energy thickness of the boundary layer - constant in logarithmic velocity profile - wavelength of laser light - kinematic viscosity - coefficient of wake function in Coles' profile Currently at LSTM, Universitat Erlangen-Nürnberg, Cauerstraße 4, W-8520 Erlangen, BRD     

Spanwise variations in nominally two-dimensional rough-wall boundary layers

Laboratory experiments have been conducted in two separate boundary layer facilities to investigate steady spanwise variations in mean velocity discovered during studies of developing flows over regular arrays of large roughness elements. Regular spanwise variation was found with a steady wavelength, moderated by the growing boundary layer, which was an integer multiple of the repeating unit of roughness. Amplitude variations greater than ±5% in the mean were found over the roughness and greater than ±10% in turbulence quantities. Due to the dominating nature of this phenomena throughout the layer, care should be taken in undertaking local measurements aimed at identifying flow variations caused by roughness heterogeneity.     

Determination of MDL 201,012 at femtomole/millilitre levels in human plasma by liquid chromatography with electrochemical detection.

A sensitive and selective liquid chromatographic method to quantitate MDL 201,012 in human plasma was developed and validated. MDL 201,012 (I), diethyl-MDL 201,012 (internal standard, II) and desmethyldiol-MDL 201,012 (masking agent, III) were isolated from basified plasma (2 mL) by solid phase extraction using Bond-Elut C-18 cartridges. Endogenous components were selectively removed prior to eluting the analytes from the sorbent. Components were separated using on-line LC column switching with a cyanopropyl precolumn and a phenyl analytical column. The analytical column effluent was monitored electrochemically at a glassy carbon electrode at a potential of +1025 mV vs. Ag/AgCl. Peak-height ratios were proportional to the amount of MDL 201,012 added to plasma over the range 125-7500 pg/mL MDL 201,012. Absolute recovery of MDL 201,012 from human plasma was > 94% across the calibration range. The minimum quantitation limit was 125 pg/mL. Assay precision (%RSD) ranged from 5.2 to 13% based on the analysis of quality control standards containing 125, 250, 500, 1000, 2500, 5000 and 7500 pg/mL MDL 201,012. Corresponding assay accuracy (% relative error) was +/- 8.5%. The method has been successfully used to quantitate MDL 201,012 in samples from acute dose tolerance studies in human volunteers.