Separation and detection of the isomeric equine conjugated estrogens, equilin sulfate and delta8,9-dehydroestrone sulfate, by liquid chromatography--electrospray-mass spectrometry using carbon-coated zirconia and porous graphitic carbon stationary phases

Equilin-3-sulfate and delta8,9-dehydroestrone-3-sulfate are two isomers found in equine conjugated estrogens that differ in structure only by the position of a double bond in the steroid B-ring. These geometric isomers were not resolved on a C18 column during the analysis of conjugated estrogen drug products by LC-MS using acetonitrile-ammonium acetate buffer as the mobile phase. While no separations of these two isomers were observed on C18 or other alkyl-bonded silica based phases using a variety of mobile phase conditions, partial separations were achieved on phenyl bonded silica phases with a resolution of 1.5 on a diphenyl phase, and baseline separations were readily achieved on two carbonaceous phases with resolutions routinely exceeding three on graphitic carbon-coated zirconia (Zr-CARB) and resolutions as high as 19 on porous graphitic carbon (Hypercarb). An examination of a selected few conjugated estrogens in the complex drug substance by LC-MS on Hypercarb is presented.     

Blue ferrocenium azurin: an organometalloprotein with tunable redox properties

A ferrocene derivative (2-[(methylsulfonyl)thio]ethylferrocene) (1) has been synthesized and incorporated into apo-azurin from Pseudomonas aeruginosa by covalent attachment to the highly conserved Cys112. The resulting artificial organometalloprotein (a protein containing organometallic compounds in the active site) has been characterized by UV-vis, electrospray mass spectrometry, and cyclic voltammetry (CV). Incorporation of 1 into azurin resulted in a higher solubility of the ferrocene group and improved stability of the ferrocenium species in aqueous solution, as shown by a more intense UV-vis absorption and a more reversible CV of the attached ferrocene group, respectively. The incorporation of 1 also increased the reduction potential of the complex from 402 to 579 mV (vs NHE), consistent with the ferrocene group being encapsulated inside the hydrophobic environment of the protein. Modulation of the reduction potential of ferrocene by residues near the secondary coordination sphere has also been demonstrated. Raising the pH from 4 to 9 resulted in a greater than 80 mV decrease in reduction potential of the protein-bound ferrocene (from 579 to 495 mV), while replacing Met121, an amino acid residue in close proximity to the ferrocene group with a positively charged Arg or negatively charged Glu, resulted in the predicted increase or decrease in reduction potential at all pH values. Similarly, substitution of Met121 with a more hydrophobic Leu raised the reduction potential. The increased solubility, stability, and tune-ability of this organometalloprotein make it an ideal choice for carrying out a number of biological reactions, such as long-range electron transfer or sensing. As an example of such applications, stoichiometric oxidation of ferrocytochrome c by the blue ferrocenium azurin was demonstrated.     

Measurement of the current-density profile and plasma dynamics in the reversed-field pinch

First measurements of the current-density profile in the core of a high-temperature reversed-field pinch are presented. The current-density profile is observed to peak during the sawtooth cycle and broaden promptly at the crash. This change in profile can be linked to magnetic relaxation and the dynamo which is predicted to drive antiparallel current in the plasma core. For high-confinement discharges, the dynamo is suppressed and the current-density profile is observed to strongly peak.     

DSA: a new internal standard for NMR studies in aqueous solution

[structure: see text] The widely used internal standard for NMR studies in aqueous solution DSS (sodium 4,4-dimethyl-4-silapentane-1-sulfonate) can interact with cationic peptides, diminishing its value for such studies. This paper introduces DSA (4,4-dimethyl-4-silapentane-1-ammonium trifluoroacetate) as a new internal standard that does not suffer from this problem.     

Electron heat transport measured in a stochastic magnetic field

New profile measurements have allowed the electron thermal diffusivity profile to be estimated from power balance in the Madison Symmetric Torus where magnetic islands overlap and field lines are stochastic. The measurements show that (1) the electron energy transport is conductive not convective, (2) the measured thermal diffusivities are in good agreement with numerical simulations of stochastic transport, and (3) transport is greatly reduced near the reversal surface where magnetic diffusion is small.     

Silicon L2,3VV Auger lineshape and oxygen chemisorption study of Pd4Si

The SiL_2,3VV Auger Lineshape for Pd₄Si was measured and found to be in good agreement with the self-fold of the Si partial density of states model calculated by Riley et al. Oxygen chemisorption altered both the Auger lineshape and the HeI photoemission spectrum, especially near the Fermi energy.     

Nonambipolar magnetic-fluctuation-induced particle transport and plasma flow in the MST reversed-field pinch

First direct measurements of nonambipolar magnetic fluctuation-induced charge transport in the interior of a high-temperature plasma are reported. Global resistive tearing modes drive the charge transport which is measured in the vicinity of the resonant surface for the dominant core resonant mode. Finite charge transport has two important consequences. First, it generates a potential well along with locally strong electric field and electric field shear at the resonant surface. Second, this electric field induces a spontaneous E x B driven zonal flow.     

Role of nonlinear coupling and density fluctuations in magnetic-fluctuation-induced particle transport

Three-wave nonlinear coupling among spatial Fourier modes of density and magnetic fluctuations is directly measured in a magnetically confined toroidal plasma. Density fluctuations are observed to gain (lose) energy from (to) either equilibrium or fluctuating fields depending on the mode number. Experiments indicate that nonlinear interactions alter the phase relation between density and magnetic fluctuations, leading to strong particle transport.