Analysis of betamethasone, dexamethasone and related compounds by liquid chromatography/electrospray mass spectrometry

A reversed-phase high-performance liquid chromatography/electrospray ionisation mass spectrometry (HPLC/ESI-MS) method has been developed to conclusively differentiate the epimers betamethasone and dexamethasone and various esterification products (betamethasone and dexamethasone 21-acetate, betamethasone and dexamethasone 21-phosphate, betamethasone 17-valerate, betamethasone 21-valerate and betamethasone 17,21-dipropionate) in counterfeit drugs. Good separation with baseline resolution of all epimers or isomers was obtained on a Zorbax Eclipse XDB or Luna C8 column, using a step gradient with mobile phases of 0.05 M ammonium acetate and acetonitrile. Betamethasones can also be distinguished by the relative abundance of their m/z 279 ion in the positive electrospray tandem mass spectra. The LC/MS or LC/MS/MS method developed was successfully applied to the analysis of drug product samples, i.e. creams and tablets.     

THE EFFECT OF ACCEPTOR GROUP VARIATION ON THE SOLVATOCHROMISM OF DONOR-ACCEPTOR FLUOROPHORES

Abstract The absorption and emission characteristics of five hydroxytetrahydrochrysenes substituted with acceptor groups (nitro, cyano, methylketone, 1° amide and methyl ester) (THC-NO₂, THC-CN, THC-COCH₃, THC-CONH₂ and THC-CO₂CH₃, respectively) were investigated in an extensive set of solvents. The order of absorption and fluorescence bathochromicity are: THC-NO₂ > THC-COCH₃ > THC-CN ≥ THC-CO₂CH₃ > THC-CONH₂ and THC-NO₂ >> THC-COCH₃ > THC-CO₂CH₃ > THC-CN > THC-CONH₂, respectively. The emission spectra of these compounds are sensitive to the solvent polarity (E_T[30] scale) in the order: THC-NO₂ > THC-COCH₃ > THC-CO₂CH₃ > THC-CONH₂ > THC-CN. The response of the emission maxima of these compounds to the solvent polarity and hydrogen-bond donor/acceptor properties (π*/α/β and acity/basity scales) was also determined. The emission energies of THC-NO₂ were most sensitive to π*, β, acity, and basity of the solvent; those of the amide were least sensitive to the solvent π*, β, and basity. The ground- and excited-state dipole moments were determined by semiempirical molecular orbital calculations and the absorption/fluorescence solvent-shift method, respectively. THC-NO₂ had the largest ground- and excitedstate moments. The ester and amide had the smallest ground- and excited-state moments, respectively. In general, unsatisfactory results were obtained for correlations of the emission and absorption energies, fluorescence solvatochromism and the ground- and excited-state dipole moments with the Hammett substituent constants of the five acceptor groups. Acceptable correlations were obtained for the absorption and emission energies and the fluorescence solvatochromism with the substituent constants if the cyano compound was excluded.     

A multi-technique surface study of the mercury(II) chalcogenide ion-selective electrode in saline media

X-ray photoelectron spectroscopy (XPS), secondary ion mass spectrometry (SIMS), rotating disc electrode-electrochemical impedance spectroscopy (RDE-EIS) and synchrotron radiation-grazing incidence X-ray diffraction (SR-GIXRD) have been used to study the response mechanism of the mercury(II) chalcogenide ion-selective electrode (ISE) in saline media. XPS and SIMS have shown that the chalcogenide surface is poisoned by silver chloride, or a mixture of silver halides, on continuous exposure to synthetic and real seawater. Significantly, the in-situ SR-GIXRD study demonstrated that electrode fouling in synthetic seawater is linked to the formation of poorly crystalline or amorphous silver chloride, and that the low level of free mercury(II) in a calibration buffer (i.e., 10(-14) M) is able to undergo metathesis with silver(II) sulfide in the membrane generating mercury(II) sulfide. Significantly, the results of this detailed surface study have shown that silver chloride fouling of the electrode is ameliorated in real seawater comprising natural organic ligands, and this has been attributed to the peptization of silver chloride by the surfactant-like nature of seawater ligands at pH 8. RDE-EIS aging studies have revealed that the chalcogenide membrane experiences a sluggish charge transfer reaction in seawater, and contrary to a previous report for a static electrode, the seawater matrix does not passivate the RDE. The results of this XPS, SIMS, RDE-EIS and SR-GIXRD study have elucidated the response mechanism of the mercury(II) ISE in saline media.     

3M Petrifilm Staph Express Count plate method for the enumeration of Staphylococcus aureus in selected types of meat, seafood, and poultry: collaborative study

The 3M Petrifilm Staph Express Count plate method was compared with AOAC Official Method 975.55 for the enumeration of Staphylococcus aureus in selected foods. Four foods--cooked, diced chicken; cured ham; smoked salmon; and pepperoni--were analyzed for S. aureus by 12 collaborating laboratories. For each food tested, the collaborators received 8 blind test samples consisting of a control sample, a low inoculation level, a medium inoculation level, and a medium inoculation level with background flora, each in duplicate. The mean log10 counts for the methods were comparable for all 4 foods. The repeatability and reproducibility variances of the 24 h Petrifilm Staph Express Count plate method were similar to those of the 72 h standard method.     

The application of diethylaminosulfur trifluoride in the synthesis of fluorinated sterols and bile acids

Methyl 3,3-(ethylenedioxy)-6β-fluoro-5β-cholan-24-oate has been prepared starting from hyodeoxycholic acid and 3,3-(ethylenedioxy)-7( and β)-fluorocholestan-22-al starting from stigmasterol, using diethylaminosulfur trifluoride as the fluorinating agent. These molecules will be used to synthesize analogs of the aminosterol antibiotic squalamine.     

Tuning cell adhesion on gradient poly(2-hydroxyethyl methacrylate)-grafted surfaces

A simple yet versatile method was developed to prepare a low-density polymerization initiator gradient, which was combined with surface-initiated atom transfer radical polymerization (ATRP) to produce a well-defined poly(2-hydroxyethyl methacrylate) (HEMA) gradient substrate. A smooth variation in film thickness was measured across the gradient, ranging from 20 A to over 80 A, but we observed a nonmonotonic variation in water contact angle. Fits of X-ray reflectivity profiles suggested that at the low graft density end, the polymer chain structure was in a "mushroom" regime, while the polymer chains at high graft density were in a "brush" regime. It was found that the "mushroom" region of the gradient could be made adhesive to cells by adsorbing adhesion proteins, and cell adhesion could be tuned by controlling the density of the polymer grafts. Fibroblasts were seeded on gradients precoated with fibronectin to test cellular responses to this novel substrate, but it was found that cell adhesion did not follow the expected trend; instead, saturated cell adhesion and spreading was found at the low grafting density region.     

Synthesis, characterization, and preliminary intramolecular energy transfer studies of rigid, emissive, rhenium-linked porphyrin dimers

The reaction of pyridyl functionalized porphyrins with Re(CO)(5)Cl in THF results in the formation of porphyrin dimers which, despite incorporation of rhenium into the assemblies, remain fluorescent. The rigid compounds provide an efficient geometry and/or orbital pathway for singlet energy transfer, rendering these compounds suitable, in principle, for the study of both through-bond and through-space energy transfer. Derivatives containing both metallated and freebase porphyrins connected via the metal corners display efficient porphyrin-porphyrin energy transfer. The photophysical properties of the assemblies have been studied by both steady-state and time-resolved fluorescence techniques, yielding approximate rates and efficiencies for porphyrin-porphyrin energy transfer.     

Neutron activation analysis of zeolite catalysts

A new method for the determination of aluminum and silicon has been developed for zeolite catalysts. In contrast to previous methods, thermal neutrons are used for the analysis of both elements, and cadmium absorbers are not needed. The silicon determination utilizes a one-hour irradiation to observe the³¹Si produced by the (n, ) reaction of³⁰Si. A 15-second irradiation is used for the²⁷Al(n, )²⁸Al reaction. The²⁸Al activity is corrected for the contribution from the²⁸Si(n,p)²⁸Al reaction by using the analyzed weight of silicon in the sample and the data for a silicon standard irradiated simultaneously with the zeolite and the aluminum standard. The quantitation limits are 0.012 g for silicon and 3.3×10^–5 g for aluminum. Sodium presents a significant interference, but this element can be removed by taking advantage of the ion exchange properties of these materials.