Large-scale purification of antisense oligonucleotides by high-performance membrane adsorber chromatography

Very high flux ion-exchange membranes were utilized for a novel purification of antisense oligonucleotides (20-mer). Strong anion-exchange membranes were produced by attaching polymeric ligands onto a microporous cellulosic matrix. The oligonucleotides purified were therapeutic single-stranded phosphorothioates deoxyribonucleotides. Although small-scale membrane devices (15 cm2) had similar resolution to traditional chromatographic columns; their throughputs were superior. Greater than a 1300-fold scale-up produced very similar purity and yields of the phosphorothionate product. Scale-up experiments were conducted with a 2 m2 surface area membrane module. These modules were easily capable of very high throughputs of 0.5 to 2 l/min. High purity and yields were achieved by both step and linear gradient elution.     

Deposition of CTAB-terminated nanorods on bacteria to form highly conducting hybrid systems

Gold nanorods and nanospheres capped with positively charged CTAB are deposited on the surface of gram-positive bacterium having negatively charged teichoic acid brushes. The deposition rate is more than an order of magnitude faster than that for peptide and nucleic acid capped nanoparticles. For the nanorods, never been reported before, the strong electrostatic attraction causes the rods to bend conformally over the curved bacterium surface in random orientations. This leads to formation of an efficient percolating cluster (i.e., low contact resistance and high radius of gyration) with 104-fold higher current at 3-fold lower coverage compared to a monolayer from nanospheres. The high monolayer conductivity at only approximately 10% coverage for nanorods has potential for fabricating a novel class of bioelectronic devices by coupling electronic "nanocircuitry" at the surface to the biological function of microorganisms.     

Hydrolysis study of organic acid anhydrides by differential thermal analysis--II. Maleic anhydride and trimellitic anhydride

Differential thermal analysis has been used to follow the hydrolysis of maleic anhydride (MA) and trimellitic anhydride (TMA). On exposure of MA to an atmosphere of 96 % relative humidity, maleic acid is produced, hydrolysis being complete in 21 hr at 22 degrees, but no hydrolysis occurs at a relative humidity of 50%. When exposed to an atmosphere of 96% relative humidity, TMA is quite stable for short periods, but hydrolyses slowly, the production of trimellitic acid being complete in 113 hr at 22 degrees.     

Photochemical production and release of gaseous NO2 from nitrate-doped water ice

Temperature-programmed NO2 emissions from frozen aqueous NaNO3 solutions irradiated at 313 nm were monitored as function of nitrate concentration and heating rate, H, above -30 degrees C. Emissions increase nonmonotonically with temperature, displaying transitions suggestive of underlying metamorphic transformations. Thus, NO2 emissions surge at ca. -8 degrees C in frozen [NO3-] > 200 microM samples warmed at H = 0.70 degrees C min(-1) under continuous irradiation, and also in the dark from samples that had been photolyzed at -30 degrees C. The amounts of NO2 released in individual thermograms, SigmaN, increase less than linearly with [NO3-] or the duration of experiments, revealing the significant loss of photogenerated NO2. The actual SigmaN proportional, variant [NO3-]1/2 dependence (at constant H) is consistent with NO2 hydrolysis: 2NO2 + H2O --> NO3- + NO2- + 2H+, overtaking NO2 desorption, even below the eutectic point (-18 degrees C for aqueous NaNO3). The increasingly larger NO2 losses detected in longer experiments (at constant [NO3-]) are ascribed to secondary photolysis of trapped NO2. The relevance of present results to the interpretation of polar NO2 measurements is briefly analyzed.     

Fast atom bombardment and collision-induced dissociation of prostaglandins and thromboxanes: Some examples of charge remote fragmentation

The mass spectra of products found by collisional activation of selected prostaglandins and thromboxanes were studied by tandem mass spectrometry as barium carboxylate salts and as carboxylate anions. Collision-induced dissociation (CID) of these closed shell ions generated by fast atom bombardment mass spectrometry reveals a wealth of structural information for these hydroxy acids. Decomposition reactions were found to be dependent upon the eicosanoid ring structure and the type of ion being studied, either positive or negative ion. The bariated carboxylate salts undergo reactions by processes that are similar to those previously characterized as charge remote mechanisms in which neutral species are lost as in thermal and photolytic decompositions. The most abundant ion is formed by loss of water from each of the hydroxyl groups present on the prostaglandin or thromboxane structure. For these multifunctionalized eicosanoids, typical patterns of decomposition emerge as characteristic of the oxygen substituents present along the carbon chain of the eicosanoid structure. The structural information obtained from the barium salts along with those from the carboxylate anions is substantially different, yet the structural information from each process is complementary. The CIDs of positive ions (metalated salts) provide structural information concerning the substituents between the carboxyl group and C₁₂ of the eicosanoid structure, whereas the decompositions of the carboxylate anions (negative ion mode) provide data concerning structure alterations of the eicosanoid structure between C₁₅ and C₂₀     

Structural dependence of thermodynamics of alkene binding to yttrium alkyl complexes and of kinetics of alkyl migration to coordinated alkenes

Agostic interactions in yttrium alkyls are structure dependent. Primary alkyl yttrium complexes have beta-CH(2) agostic interactions at low temperature, but a shift toward alpha-agostic interactions occurs on warming. For the more crowded beta-disubstituted yttrium alkyls, an alpha-CH(2) agostic interaction is seen. The thermodynamics of alkene binding to the primary alkyl yttrium complex Cp(2)YCH(2)CH(2)CH(CH(3))(2) (2) depend strongly on the structure of the alkene. A single allylic substituent on the alkene has a small effect on alkene binding, but a second allylic substituent has a large destabilizing effect. Propene binding to yttrium alkyls is largely independent of the nature of the alkyl ligand. Equilibrium constants for propene binding to n-, gamma-substituted, beta-substituted, and secondary alkyl yttrium complexes are similar. The rate of migration of an alkyl group to a coordinated alkene depends strongly on the structure of the alkyl group: n-alkyl approximately gamma-substituted > beta-substituted > alpha-substituted. The approximately 200-fold slower insertion of propene into Cp(2)YCH(2)CH(CH(3))(2) (6) than that into Cp(2)YCH(2)CH(2)CH(CH(3))(2) (2) is therefore due to kinetically slow migration of the beta-disubstituted alkyl group of 6 and not to differences in the equilibrium binding of propene. Processes related to chain transfer and site epimerization at the metal center are also reported.     

Urine drug testing for opioids,cocaine, and metabolites by direct injection liquid chromatography/tandem mass spectrometry

A sensitive and specific liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the simultaneous quantification of opioids, cocaine, and metabolites in urine was developed and validated. A 10-microL aliquot of urine was injected directly onto the LC/MS/MS system. The lack of sample preparation substantially reduced total analysis time. Separation was performed by reversed-phase chromatography with gradient elution for all analytes in 26 min. Atmospheric pressure chemical ionization (APCI) was a rugged and efficient ionization technique for basic drugs. Identification and quantification was based on selected reaction monitoring (SRM). Calibration, with deuterated internal standards, was performed by linear regression analysis (weighting factor 1/x). Limits of quantitation (LOQ) were established between 10-100 ng/mL and linearity was obtained up to a maximum of 10 000 ng/mL with an average correlation coefficient (R(2)) > 0.99. Analytical validation criteria for specificity, precision, accuracy, dilution integrity, matrix effect, and stability were fulfilled. The method proved to be simple and time efficient, and was applicable for illicit drug use monitoring and methadone treatment compliance in clinical research projects at the National Institute on Drug Abuse (NIDA).     

An ENDOR and DFT analysis of ‘solvatochromic’ effects in an oxovanadium (IV) complex

The ‘solvatochromic’ effects of a vanadyl salen complex [VO^IV(salen)] in frozen solution was studied by ENDOR and DFT calculations. In a non-coordinating solvent (dichloromethane), both ENDOR and DFT were in excellent agreement on the expected square pyramidal structure, where V^IV=O is positioned out of the equatorial ligand xy-plane (as determined from calculated VH distances). In a coordinating solvent (dimethylformamide), a subtle perturbation from the square pyramidal structure was observed, suggesting that DMF coordinates trans to the vanadyl oxo-ligand, pulling V^IV=O back into the ligand plane. The axial coordination of DMF was confirmed by ENDOR and in the DFT optimised [VO^IV(salen)]–DMF complex.     

Synthesis of an influenza neuraminidase inhibitor intermediate via a highly diastereoselective coupling reaction

[reaction: see text]. A highly diastereoselective coupling reaction between TBSOP (3) and trityl sulfenimine 4 was developed which provided influenza neuraminidase inhibitor intermediate 7 in 80% yield and >99% de after crystallization. The reaction was shown to be reversible with the high diastereoselectivity resulting from a favorable H-bonding interaction in the major diastereomer.     

A chiral, heterometallic metal-organic framework derived from a tris(chelate) coordination complex

A novel tris(chelate) metalloligand has been used to synthesize a chiral, heterometallic metal-organic framework that is robust to solvent removal and shows selective uptake of nitroaromatic compounds.