Separation of triphenyl atropisomers of a pharmaceutical compound on a novel mixed mode stationary phase: a case study involving dynamic chromatography, dynamic NMR and molecular modeling.

Analysis of atropisomers is of considerable interest in the pharmaceutical industry. For complex chiral molecules with several chiral centers hindered axial rotation can lead to formation of interconverting diastereomers that should be separable on achiral stationary phases. However, achieving the actual separation may be difficult as the on-column separation speed must match or be faster then the rate of isomer interconversion. Often, this requirement can be satisfied by using low-temperature conditions and by improving selectivity via use of chiral stationary phases. In the current study, we present an alternative approach utilizing an Obelisc R column, a novel mixed mode stationary phase that provided acceptable separation of triphenyl atropisomers inside a conventional HPLC temperature range. The separation was investigated under various chromatographic conditions. The interconversion chromatograms exhibited classic peak-plateau-peak behavior indicating the simultaneous atropisomer separation and interconversion. The elution profiles were integrated in order to deconvolute the peak areas of the "pure" (non-exchanged) and interconverted species; these data were used to obtain kinetic information. Analysis of retention data rendered thermodynamic information on the mechanism of retention and selectivity. Chromatographic kinetic data were complemented with variable-temperature NMR and molecular modeling studies, which provided additional support and insights into the energetics of the interconversion process.

Chromatographic behavior of ion pair enantiomers of dansyl leucine cyclohexylammonium salt on a beta-cyclodextrin stationary phase and the effect of a competitive-binding mobile phase additive.

The separation of dansyl leucine enantiomers on a beta-cyclodextrin stationary phase is significantly complicated by the association of the amino acid with its cyclohexylammonium counter ion, in a mobile phase of 80:20 (v/v) methanol-water. This produces very unusual chromatography, with two partially superimposed peaks observed for each enantiomer at lower column temperatures. The peak shape is attributed to the irreversible, oncolumn conversion of the ion pair (I) to the free, protonated (neutral) dansyl amino acid (II+H). Increasing the ionic strength of the mobile phase greatly improves the chromatography by transforming the solute species to enantiomers of II (the anionic, free amino acid). Van't Hoff plots are constructed for both species I and II (under different mobile phase conditions) to provide thermodynamic insight into the major enantioselective driving forces of separation. The chiral discrimination of the stationary phase is found to be primarily enthalpically driven for both solutes. Finally, 1-adamantanecarboxylic acid (ACA) is investigated as a solute-competitive mobile phase additive to intentionally block the hydrophobic cyclodextrin cavities on the stationary phase. By varying the concentration of ACA additive in the mobile phase, control over the retention and chiral recognition of the stationary phase is demonstrated.

Tautomer interconversion of 2,4-pentanedione during gas chromatography on an oxidized cyano-modified capillary column.

The tautomerization of 2,4-pentanedione (acetylacetone) is examined on a microbore column containing an acid-modified stationary phase made by oxidizing a commercially available cyano-modified column. This stationary phase is found to provide separation of the two tautomers, which allows the kinetic and thermodynamic properties of the on-column interconversion to be investigated. The enol-to-keto tautomerization is found to occur primarily in the stationary phase, being enthalpically driven. By treating the column as a reactor, the interconversion is investigated as a function of temperature. Monitoring the loss of the more gas-stable 'enol' tautomer makes it possible to extract an energy of activation for the net tautomerization (42.7 kj/mol), because the reaction is found to obey pseudo first-order kinetics. Simple peak-shape analysis of the major component (enol), which is used commonly in treatments of peak tailing, provides insight into the nature of the retention processes of the two tautomers as well as information on chromatographic optimization.

A simple quantitative FT-IR approach for the study of a polymorphic transformation under crystallization slurry conditions.

The pharmaceutical compound (2R,3S)-2-([(1R)-1-[3,5-bis(trifluoromethyl)phenyl]ethyl]oxy)-3-(4-fluorophenyl)morpholine hydrochloride (denoted here as Compound X), has been found to crystallize in at least two polymorphic forms. Using only two frequencies (1009 and 1058 cm(-1)) in the infrared, a linear (R=0.998) calibration plot, consisting of the ratio of the peak absorbances plotted against polymorph concentration, was constructed. This plot allowed the quantification of binary mixtures of polymorphs ranging from <3 to approximately 100 wt% Form II in Form I. Spectra were acquired in transmission mode using mineral oil (Nujol) mull sample preparation, for reasons of compatibility with wet cake and slurry samples. The transformation of the less thermodynamically stable polymorph (Form II) to the more stable form (Form I), in stirred methyl isobutyl ketone (MIBK) slurries, was monitored spectroscopically as a function of time. Performing the experiment at various temperatures allowed the energy of activation for the process to be estimated (42 kJ/mol).

Preparation of polyetherol-appended sulfur porphyrazines and investigations of peripheral metal ion binding in polar solvents.

We describe the complete synthesis and characterization of a new family of peripherally functionalized porphyrazines (pz's) with four, three, or two (in a trans conformation) bis[thioethoxy(ethoxy)ethanol] moieties appended at the pyrroles. These "polyetherol" groups serve as weak exocyclic binding sites for a number of metal ions and also provide solubility of the pz's in low molecular-weight alcohols and water. Electronic spectra of the modified porphyrazines exhibit distinct changes in the visible region (both absorbance and fluorescence) in response to treatment with Ag+, Pb2+, Cd2+, Cs+, and Ni2+ in solution. Such properties make these compounds intriguing candidates for incorporation into the transducer layers in optically based chemical sensors.

Sol-gel-based, planar waveguide sensor for water vapor.

A water vapor sensor based on a combination of sol-gel processing and planar optical waveguide technologies has been developed. The indicator erythrosin B was entrapped in a thin sol-gel film (thickness ∼100 nm) prepared from methyltriethoxysilane, dimethyldiethoxysilane, and tetraethoxysilane. This dye exhibits an increase in absorbance in the presence of liquid or gaseous water. The sol-gel layer containing the dye was deposited onto a sol-gel-derived, single-mode planar waveguide. Outcoupled light intensity measurements (at 514.5 nm) over a range of water vapor concentrations (in a nitrogen gas stream) yielded a response over a wide range of relative humidity (<1-∼70%) at room temperature. Response and reversal times were less than 1 min, which may make this sensor attractive for real-time monitoring applications.