A Simple and Efficient Approach for Estimating Recovery of a Preparative Reversed Phase HPLC Purification

The widely applied reversed phase high-performance liquid chromatography (RP-HPLC) is an indispensable purification technique in drug discovery. During drug discovery, recovery was usually calculated based on the weight of the purified product after drying over the weight of the crude material multiplied by the assumed purity from HPLC/UV area percent of the product. Such a purity assumption can be off significantly when the crude material contains water, solvents, other UV-inactive impurities and inorganic salts. In this paper, we report a simple and efficient way to estimate recovery of preparative HPLC purification process. It is based on the ratio of the HPLC/UV peak area measured for the product in the crude solution and that in the final collected fraction with both accounted for their volumes. This approach eliminates not only the need for drying of the collected fraction to calculate recovery but also the inaccuracy associated with the true content in the crude sample using the traditional method. A systematic study was conducted to verify this method using caffeine mixed with various UV-active and -inactive impurities. The calculated recoveries using this approach were found to be consistent within 4% with the true recoveries based on dry weight estimation. The approach has been successfully applied for our in-house purifications. Furthermore, the approach was extended to library purifications, where in many cases heart-cutting the desired peaks is used to meet the purity requirements.

     

Generalized fabrication of two-dimensional non-close-packed colloidal crystals

In this paper we report a generalized templating approach for fabricating wafer-scale, two-dimensional, non-close-packed (ncp) colloidal crystals. Polymer nanocomposites consisting of monolayer ncp colloidal crystals prepared by a spin-coating process are used as sacrificial templates. After removal of the colloidal silica templates, the voids in the polymer matrix are infiltrated with other materials. By plasma-etching the polymer matrix, wafer-scale ncp colloidal crystals from a variety of functional materials can be made. This technique is scalable and compatible with standard microfabrication. Two-component colloidal arrays with complex micropatterns can also be fabricated by combining microfabrication with this templating approach. Normal-incidence reflectivity spectra of replicated titania ncp arrays agree well with theoretical prediction using Scalar Wave Approximation.     

High-temperature shock tube measurements of methyl radical decomposition

We have studied the two-channel thermal decomposition of methyl radicals in argon, involving the reactions CH3 + Ar --> CH + H2 + Ar (1a) and CH3 + Ar --> CH2 + H + Ar (1b), in shock tube experiments over the 2253-3527 K temperature range, at pressures between 0.7 and 4.2 atm. CH was monitored by continuous-wave, narrow-line-width laser absorption at 431.1311 nm. The collision-broadening coefficient for CH in argon, 2gamma(CH-Ar), was measured via repeated single-frequency experiments in the ethane pyrolysis system behind reflected shock waves. The measured 2gamma(CH-Ar) value and updated spectroscopic and molecular parameters were used to calculate the CH absorption coefficient at 431.1311 nm (23194.80 cm(-1)), which was then used to convert raw traces of fractional transmission to quantitative CH concentration time histories in the methyl decomposition experiments. The rate coefficient of reaction 1a was measured by monitoring CH radicals generated upon shock-heating highly dilute mixtures of ethane, C2H6, or methyl iodide, CH3I, in an argon bath. A detailed chemical kinetic mechanism was used to model the measured CH time histories. Within experimental uncertainty and scatter, no pressure dependence could be discerned in the rate coefficient of reaction 1a in the 0.7-4.2 atm pressure range. A least-squares, two-parameter fit of the current measurements, applicable between 2706 and 3527 K, gives k(1a) (cm(3) mol(-1) s(-1)) = 3.09 x 1015 exp[-40700/T (K)]. The rate coefficient of reaction 1b was determined by shock-heating dilute mixtures of C2H6 or CH3I and excess O2 in argon. During the course of reaction, OH radicals were monitored using the well-characterized R(1)(5) line of the OH A-X (0,0) band at 306.6871 nm (32606.52 cm(-1)). H atoms generated via reaction 1b rapidly react with O2, which is present in excess, forming OH. The OH traces are primarily sensitive to reaction 1b, reaction 9 (H + O2 --> OH + O) and reaction 10 (CH3 + O2 --> products), where the rate coefficients of reactions 9 and 10 are relatively well-established. No pressure dependence could be discerned for reaction 1b between 1.1 and 3.9 atm. A two-parameter, least-squares fit of the current data, valid over the 2253-2975 K temperature range, yields the rate expression k(1b) (cm(3) mol(-1) s(-1)) = 2.24 x 10(15) exp[-41600/T (K)]. Theoretical calculations carried out using a master equation/RRKM analysis fit the measurements reasonably well.     

Existence and Weyl’s law for spherical cusp forms

Let G be a split adjoint semisimple group over and a maximal compact subgroup. We shall give a uniform, short and essentially elementary proof of the Weyl law for cusp forms on congruence quotients of . This proves a conjecture of Sarnak for -split groups, previously known only for the case G = PGL(n). The key idea amounts to a new type of simple trace formula. Received: April 2005 Revision: June 2006 Accepted: October 2006     

Spectrofluorometric studies on the binding interaction of bioactive imidazole with bovine serum albumin: a DFT based ESIPT process

Bioactive imidazole derivatives were synthesized and characterized by NMR spectra, mass and CHN analysis. An excited state intramolecular proton transfer (ESIPT) process in hydroxy imidazole has been studied using emission spectroscopy. In hydrocarbon solvent, the tautomer emission predominates over the normal emission and in alcoholic solvent like ethanol; a dramatic enhancement of normal emission is observed which was due to increased solvation. DFT calculation on energy, charge distribution of the rotamers in the ground and excited states of the imidazole derivative were performed and discussed. PES calculation indicates that the energy barrier for the interconversion of two rotamers is too high in the excited state than in the ground state. The interaction between bioactive imidazole derivative and bovine serum albumin (BSA) was investigated.     

Solvatochromic analysis of some N-nitroso oxime derivatives--Taft and Catalan approach

Some novel N-nitroso oxime derivatives were synthesized and characterized by (1)H, (13)C, (1)H-(1)H and (1)H-(13)C COSY NMR spectra. The spectra of all these N-nitroso oximes reveal the presence of two isomers labelled as E (-NOH group is anti to N-N=O moiety) and Z (-NOH group is syn to N-N=O moiety) in solution and the coupling constants ruled out the possibility of normal chair conformation. From the theoretical studies and coupling constant values it was found that both E (major) and Z (minor) isomers of N-nitroso oximes exist as an equilibrium mixture of CA and boat conformation (B(1)) and this was also supported by DFT calculation. The photophysical properties of these oxime derivatives were studied and the observed lower fluorescence quantum yield may be due to an increase in the non-radiative deactivation rate constant. This is attributed due to the presence of non-chair conformation of N-nitroso oxime derivatives.     

Photophysical Properties of Novel Picrate Derivatives – Solvent Effect

H and ¹³C NMR spectra were recorded for some novel picrate derivatives derived from 3,3-dimethyl-2,6-diarylpiperidin-4-ones and 3-benzyl-2,6-diarylpiperidin-4-one. The photophysical properties of the picrate derivatives were studied in several solvents, which include a wide range of apolar, polar and protic media. The observed lower fluorescence quantum yield may be due to an increase in the non-radiative deactivation rate constant. This is attributed due to the presence of increased electrostatic interaction between N-protonated piperidone ring and picryl anion ring so that the picryl anion ring lies perpendicular to the plane of the N-protonated piperidone ring i.e., non co-planarity. Such a geometrical change in the excited state leads to an important Stokes shift, reducing the reabsorption and reemission effects in the detected emission in highly concentrated solutions. The higher fluorescence quantum yield of the picrate derivatives are observed in polar media.     

Large-eddy Simulation of Motored Flow in a Two-valve Piston Engine: POD Analysis and Cycle-to-cycle Variations

Large-eddy simulation (LES) has been performed for a single-cylinder, two-valve, four-stroke-cycle piston engine through 70 consecutive motored cycles. Initial comparisons of ensemble-averaged velocity fields have been made between LES and experiment, and proper orthogonal decomposition (POD) has been used to analyze the complex in-cylinder turbulent flows. Convergence of POD modes has been quantified, several POD variants have been explored, and sensitivity of results to analyzing different subsets of engine cycles has been studied. In general, it has been found that conclusions that were drawn earlier from POD analysis of a simplified non-compressing piston-cylinder assembly with a fixed valve carry over to the much more complex flow in this motored four-stroke-cycle engine. For the cases that have been examined, the first POD mode essentially corresponds to the ensemble-averaged mean velocity. The number of engine cycles required to extract converged POD modes increases with mode number, and varies with phase (piston position). There is little change in the lower-order phase-invariant POD modes when as few as 24 phases per cycle (30° between samples) are used, and complex 3-D time-dependent in-cylinder velocity fields through full engine cycles can be reconstructed using a relatively small number of POD modes. Quantification of cycle-to-cycle variations and insight into in-cylinder flow dynamics can be extracted through analysis of phase-invariant POD modes and coefficients.