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.

Kinetic analysis and subambient temperature on-line on-column derivatization of an active aldehyde.

The chromatographic analysis of aldehydes under typical reversed-phase conditions may be a challenging task due to an equilibrium process leading to the formation of a gem diol species regardless of acidic or basic conditions. Initially, a reversed-phase HPLC gradient elution was developed to determine the amount of a n acetylenic aldehyde in a reaction mixture. Significant fronting was observed under acidic and basic conditions even at -5 degrees C. In order to circumvent this problem, a reversed-phase HPLC gradient method on a C18 column at subambient temperature was developed using diethylamine as a mobile phase additive for on-line on-column derivatization of the aldehyde moiety. The on-line on-column reaction rate for the derivatization of the aldehyde with diethylamine was determined as a function of column temperature. An Arrhenius plot was constructed and the activation energy was calculated. The chromatographic behavior of the derivatized acetylenic aldehyde and products formed in-situ in the chromatographic system were studied at various temperatures ranging from -10 to 60 degrees C. It was found that the reaction products could be controlled by adjusting the column temperature. Different reaction pathways were identified as a function of temperature. The products and the reaction pathways were characterized by NMR, LC-MS and UV spectra.

Mechanistic aspects of chiral discrimination on an amylose tris(3,5-dimethylphenyl)carbamate.

The separation of [2R-[2alpha(R*),3alpha]]-5-[[2-[1-[3,5-bis-(trifluoromethyl)phenyl]ethoxy]-3(S)-4-fluorophenyl)4-morpholinyl]-methyl]-N,N-dimethyl-1H-1,2,3-triazole-4-methanamine hydrochloride from its enantiomer was achieved on an amylose tris-3,5-dimethylphenyl carbamate stationary phase. The retention of the enantiomers is dominated by weak hydrogen bonds while the enantioselectivity is governed by other kinds of interactions, e.g., inclusion in the amylose carbamate chains. Van't Hoffplots of 1nalpha vs. reciprocal temperature were non-linear and could be divided into two linear regions. One region at low temperature (5 degrees C- approximately 20 degrees C) and another one between 25 degrees C-70 degrees C with the change in slope occurring between 16 degrees C and 20 degrees C. DSC experiments suggested that the behavior can be attributed to breakage of H-bonds triggering a conformational change. Molecular simulation indicated a correlation between the interaction energies and the elution order obtained experimentally. The most retained enantiomer (R,R,S-enantiomer) interacts with the stationary phase through a hydrogen bond between the triazole proton and the C=O groups of the stationary phase, as well as through an inclusion in the cleft of the stationary phase. The other enantiomer exhibits a bifurcated H-bond between the triazolic proton and the C=O groups of the stationary phase leading to a less stable complex.

Thin-layer chromatography--a useful technique for the separation of enantiomers.

Separation of enantiomers has become a well-established technique in many fields of science over the last decade. Unfortunately, even though there are a large number of chiral stationary phases able to perform enantiomeric separation, there is still a great deal of trial and error in developing a method for the separation of enantiomers. Thin-layer chromatography is a very versatile technique, which has brought much advancement in various fields of science. The simplicity of the technique makes it amenable for separation of enantiomers. This paper will present a review of the literature concerning separation of enantiomers. Because of the process of trial and error present in developing a chiral separation method, this paper also presents the mechanism underlying each form of separation. Thus, the methods are presented according to the main mechanism governing the particular separation.

Adsorption Characterization of Two Clay Minerals Society Standard Kaolinites.

The kaolinite samples KGa-1b and KGa-2 are used as standards by the Clay Minerals Society of America. Recently a comparative study of these samples was carried out using atomic force microscopy (AFM) and X-ray diffraction methods (XRD). The aim of the current work was to characterize the kaolinite standards by high resolution nitrogen adsorption and to investigate how their surface heterogeneity changes upon sample cleaning. Copyright 1998 Academic Press.

Surface and Structural Properties of Novel Titanium Phosphates

Two novel fibrous framework titanium phosphates having a similar formula, Ti2O(PO4)2·H2O, and a layered titanium phosphate, Ti2O3(H2PO4)2·2H2O, were synthesized and their properties were characterized with physicochemical methods, such as elemental analysis, thermogravimetry, nitrogen adsorption, X-ray diffraction, and 31P solid-state NMR. Nitrogen adsorption isotherms were measured on synthesized titanium phosphates in order to characterize their surface and structural properties. The specific surface area and total pore volume of the titanium phosphates studied were determined. The low-pressure nitrogen adsorption isotherms were used to evaluate surface heterogeneity of the synthesized materials by employing an advanced numerical procedure based on the regularization method.