Green HPLC Enantioseparation of Chemopreventive Chiral Isothiocyanates Homologs on an Immobilized Chiral Stationary Phase Based on Amylose tris-[(S)-α-Methylbenzylcarbamate].

Sulforaphane is a chiral phytochemical with chemopreventive properties. The presence of a stereogenic sulfur atom is responsible for the chirality of the natural isothiocyanate. The key role of sulfur chirality in biological activity is underscored by studies of the efficacy of individual enantiomers as chemoprotective agents. The predominant native (R) enantiomer is active, whereas the (S) antipode is inactive or has little or no biological activity. Here we provide an enantioselective high-performance liquid chromatography (HPLC) protocol for the direct and complete resolution of sulforaphane and its chiral natural homologs with different aliphatic chain lengths between the sulfinyl sulfur and isothiocyanate group, namely iberin, alyssin, and hesperin. The chromatographic separations were carried out on the immobilized-type CHIRALPAK IH-3 chiral stationary phase with amylose tris-[(S)-methylbenzylcarbamate] as a chiral selector. The effects of different mobile phases consisting of pure alcoholic solvents and hydroalcoholic mixtures on enantiomer retention and enantioselectivity were carefully investigated. Simple and environmentally friendly enantioselective conditions for the resolution of all chiral ITCs were found. In particular, pure ethanol and highly aqueous mobile phases gave excellent enantioseparations. The retention factors of the enantiomers were recorded as the water content in the aqueous-organic modifier (methanol, ethanol, or acetonitrile) mobile phases progressively varied. U-shaped retention maps were generated, indicating a dual and competitive hydrophilic interaction liquid chromatography (HILIC) and reversed-phase liquid chromatography retention mechanism on the CHIRALPAK IH-3 chiral stationary phase. Finally, experimental chiroptical studies performed in ethanol solution showed that the (R) enantiomers were eluted before the (S) counterpart under all eluent conditions investigated.

Optimization of Liquid Crystalline Mixtures Enantioseparation on Polysaccharide-Based Chiral Stationary Phases by Reversed-Phase Chiral Liquid Chromatography.

Enantioseparation of nineteen liquid crystalline racemic mixtures obtained based on (R,S)-2-octanol was studied in reversed-phase mode on an amylose tris(3-chloro-5-methylphenylcarbamate) (ReproSil Chiral-MIG) and a cellulose tris(3,5-dichlorophenylcarbamate) (ReproSil Chiral-MIC). These polysaccharide-based chiral stationary phase (CSP) columns for High-Performance Liquid Chromatography (HPLC) were highly effective in recognizing isomers of minor structural differences. The mobile phase (MP), which consists of acetonitrile (ACN)/water (H2O) at different volume ratios, was used. The mobile phases were pumped at a flow rate of 0.3, 0.5, or 1 mL·min-1 with a column temperature of 25 °C, using a UV detector at 254 nm. The order of the elution was also determined. The chromatographic parameters, such as resolution (Rs), selectivity (α), and the number of theoretical plates, i.e., column efficiency (N), were determined. The polysaccharide-based CSP columns have unique advantages in separation technology, and this study has shown the potential usefulness of the CSP columns in separating liquid crystalline racemic mixtures belonging to the same homologous series.

Exploring a Lux® i-Amylose-3 column in normal phase and polar-organic mode for chiral separation of cathinone derivatives and pyrovalerones using high-performance liquid chromatography.

Each year, new psychoactive substances appear on the global drug market leading to constant changes. Most of these compounds with stimulating effect possess a chiral center, thus leading to two enantiomers with presumably different pharmacological properties. Among them, synthetic cathinones, often misleadingly traded as "bath salts," play an important role. There is little knowledge about the distinct effect of the enantiomers. The aim of this study was to test a commercially available Lux® i-Amylose-3 column by HPLC-UV for enantiorecognition of cathinone derivatives. Overall, 80 compounds were tested in normal phase mode, where 75 substances were separated under initial conditions. After method optimization, at least partial separation was achieved for the remaining compounds. The same set of substances was measured in polar-organic mode, where 63 analytes were resolved into their enantiomers under initial conditions with very short retention times. Both modes showed complementary results for the individual compounds. Furthermore, the tested methods proved to be suitable for differentiation of positional isomers, which can be useful for drug checking programs. All measurements were carried out under isocratic conditions, and intraday and interday repeatability tests were performed.

Enantioseparation of selected chiral agrochemicals by using nano-liquid chromatography and capillary electrochromatography with amylose tris(3­chloro-5-methylphenylcarbamate) covalently immobilized onto silica.

In this paper enantiomers of selected chiral agrochemicals representing various structural classes were separated by using nano-liquid chromatography (nano-LC) and capillary electrochromatography (CEC) employing a capillary column packed with silica particles containing immobilized amylose tris(3­chloro-5-methylphenylcarbamate) (i-ADMPC) as a chiral selector (CS). Special attention was paid to peak dispersion in nano-LC and CEC instruments used in order to make comparison between these two techniques more reliable. Enantioseparations were studied utilizing methanol (MeOH) or acetonitrile-water (ACNH2O), both containing 5 mM of ammonium acetate as the mobile phases (MPs). The tested chiral stationary phase (CSP), containing 20% (w/w) of the neutral CS onto native silica, allowed the generation of sufficiently strong electroosmotic flow (EOF) to observe separation of enantiomers of studied agrochemicals in a reasonable time also in CEC mode. Modestly higher efficiencies and enantioresolutions were obtained in CEC than in nano-LC. Just a moderate preference of CEC over nano-LC in this particular study can be explained with a significant mass transfer resistance through the CSP that is caused due to high content of the CS in CSP.

A protocol to replace dedication to either normal phase or polar organic mode for chiral stationary phases containing amylose tris(3,5-dimethylphenylcarbamate).

Alteration of the enantiorecognition ability of polysaccharide-based chiral columns in the shipping normal phase (NP) eluent after exposition to polar organic (PO) mode eluents can be conceived as an incomplete hysteresis cycle. Non-standard solvents provide a solution to overcome this issue with immobilized stationary phases, but a procedure was missing so far to regenerate coated stationary phases from the altered state. Recent results with alcohol mixtures within the PO mode showed that an appropriate order of standard solvents may also be suitable to complete hysteresis. Using an analogous approach, a simple protocol was established to regenerate the original NP retentions on various stationary phases containing amylose tris(3,5-dimethylphenylcarbamate) (ADMPC) chiral selector after the change induced by flushing with 2-propanol or ethanol. The members of a chemically diverse compound set indicated that alterations in retentions and selectivities using different brands and types of ADMPC-based stationary phases can be quite different, but the recovery of the original state was very good for all of them. The proposed protocol eliminates the need of the costly dedication of a chiral column with ADMPC selector to either NP or PO mode. Furthermore, the limits of the alcohol content in the mobile phase compositions where the system is free of hysteresis were determined.

Analytical Separation of Closantel Enantiomers by HPLC.

Closantel is an antiparasitic drug marketed in a racemic form with one chiral center. It is meaningful to develop a method for separating and analyzing the closantel enantiomers. In this work, two enantiomeric separation methods of closantel were explored by normal-phase high-performance liquid chromatography. The influences of the chiral stationary phase (CSP) structure, the mobile phase composition, the nature and proportion of different mobile phase modifiers (alcohols and acids), and the column temperature on the enantiomeric separation of closantel were investigated in detail. The two enantiomers were successfully separated on the novel CSP of isopropyl derivatives of cyclofructan 6 and n-hexane-isopropanol-trifluoroacetic acid (97:3:0.1, v/v/v) as a mobile phase with a resolution (Rs) of about 2.48. The enantiomers were also well separated on the CSP of tris-carbamates of amylose with a higher Rs (about 3.79) when a mixture of n-hexane-isopropanol-trifluoroacetic acid (55:45:0.1, v/v/v) was used as mobile phase. Thus, the proposed separation methods can facilitate molecular pharmacological and biological research on closantel and its enantiomers.

Enantioselective separation of nonsteroidal anti-inflammatory drugs with amylose tris(3-chloro-5-methylphenylcarbamate) stationary phase in HPLC with a focus on enantiomeric quality control in six pharmaceutical formulations containing racemic mixtures or single stereoisomers.

In the present study, an accurate, rapid, and simple chiral HPLC-UV method with amylose tris(3-chloro-5-methylphenylcarbamate) as stationary phase was developed and applied for enantiomeric determination of six nonsteroidal anti-inflammatory drugs (NSAIDs) in the commercial pharmaceutical formulations, including (R,S)-ibuprofen, S-ibuprofen, (R,S)-ketoprofen, S-ketoprofen, S-naproxen, and (R,S)-loxoprofen sodium. Experiments on the influence of mobile phase composition, proportion of organic modifier, percentage of acid additives, and column temperature on enantioseparation were conducted to obtain the best separation condition. It was indicated that one mobile phase simply composed of acetonitrile-water (0.1% formic acid, v/v) at the proportion of 50:50 (v/v) with a flow rate of 0.6 ml/min at 22°C could simultaneously provide the excellent enantiomeric resolutions for all selected NSAIDs, which made the enantioseparation process more applicable and operable. The newly developed method was then applied for determination of NSAID enantiomers in pharmaceutical formulations containing racemic mixtures or single stereoisomers. Calibration curve of each enantiomer at the concentration of 5.0-100 ug/ml showed good linearity with the correlation coefficient above 0.9996. Satisfactory recovery (96.54-101.54%), good intra-day precision (RSD 0.52-1.46%), and inter-day precision (RSD 0.13-1.09%) were also obtained. The newly developed method was then applied for determination of NSAID enantiomers in pharmaceutical formulations containing racemic mixtures or single stereoisomers. Quantitative results of the commercial capsules and tablets demonstrated that the difference between the declared and measured values did not exceed 1.52%.

Controlling plasticizer migration based on crystal structure and micromorphology in propionylated starch-based food packaging nanocomposites.

Migration of additives is an important issue for proper application in food packaging. In this work, propionylated waxy and normal starch-based nanocomposites (PW-N and PN-N) with two different amylopectin content were immersed in distilled water, and structural changes and migration mechanism of plasticizer (triacetin) were discussed in detail. Results showed that when immersion time was prolonged to 150 h, small crystals of PN-N disappeared, and amorphous structures formed gradually in PW-N and PN-N. Exfoliated structures still remained in PW-N with prolonged immersion time, while exfoliated structures gradually formed from intercalated ones in PN-N, and the peak representing d001 (d-spacing) at q = 1.70 nm-1 faded. The migration mechanism of triacetin obeyed the first-order kinetic model and Fick's law; furthermore, in comparison with PW-N, PN-N showed a larger diffusion coefficient (D2 = 12.13 µm2·h-1). These results contributed to expanding the application of starch-based nanocomposites in future environmentally friendly food packaging.

Effect of 2-propanol content on solute retention mechanisms determined using amylose tris(3,5-dimethylphenylcarbamate) chiral stationary phase under normal- and reversed-phase conditions.

The electrostatic interactions between chiral solutes and polysaccharide (PS)-based chiral selectors are the key to achieving chiral recognition; however, PS-based sorbents, derivatized of phenyl moieties, can exhibit considerably non-polar characteristics, and they are also useful for the separation of enantiomers in the reversed-phase mode. In this study, an immobilized amylose 3,5-dimethylphenylcarbamate-based sorbent was used to investigate the balance between electrostatic interactions and solvophobic interactions, with complementary effects on solute retention behavior when the isopropanol (IPA) concentration was altered. It was proposed that in both normal- and reversed-phase modes, information on the retention mechanisms could be obtained by observing the curvature of the logarithm of the retention factor versus the logarithm of the IPA concentration, and the slope values of the curves were related to the number of displaced IPA molecules upon solute adsorption. Using the proposed model and the two-site adsorption model, the retention behaviors of pantolactone (PL) enantiomers in both normal- and reversed-phase modes were investigated. The PL-sorbent interactions were classified into four types: electrostatic/enantioselective, electrostatic/nonselective, solvophobic/enantioselective, and solvophobic/nonselective. At IPA concentrations below 50 vol.% in n-hexane, the retention behaviors of PL were dominated by electrostatic/enantioselective sites, whereas at IPA concentrations beyond 50 vol.%, the solvophobic interactions of PL-sorbent were strengthened and mostly nonselective. By contrast, in the reversed-phase mode, a reverse in the enantiomeric elution order of PL was observed at 10 vol.% IPA, and considerably different enantioselectivity behaviors were found below and above 20 vol.%, indicating an abrupt change in the sorbent molecular environment. At IPA concentrations beyond 40 vol.%, the presence of PL-sorbent electrostatic interactions enhanced chiral recognition.

Supercritical fluid chromatography and liquid chromatography for isomeric separation of a multiple chiral centers analyte.

The development of a chiral separation strategy has always been a challenge of crucial importance, particularly in the pharmaceutical field. Chromatographic methods have become popular, particularly High Performance Liquid Chromatography and Supercritical Fluid Chromatography from a preparative scale point of view. A bioactive compound bearing three stereogenic centers was entrusted in our laboratory and the aim of this work was to obtain the complete resolution of the eight stereoisomers. Nine different polysaccharide-based columns were tested in SFC under various carbon dioxide-based mobile phases. The use of a single chiral column Lux Cellulose-2 under 30% 2-PrOH in carbon dioxide, at a flow-rate of 1 mL/min, column temperature of 40°C, 120 bar outlet pressure allowed the obtention of eight peaks. To further improve the resolution of the two last isomers, two columns were serially coupled . The results obtained with the six different combinations are discussed. The tandem column supercritical fluid chromatography has demonstrated to be a useful technique to resolve the eight stereoisomers on Lux Cellulose-2//Cellulose-2 tandem of coupled columns with 30% 2-PrOH in carbon dioxide, at a flow-rate of 1 mL/min, column temperature of 40°C and 120 bar outlet pressure, despite a long analysis time. In order to compare the two methods (i.e supercritical and liquid), chiral liquid chromatography under polar aqueous-organic mode, polar organic mode and normal-phase mode, was implemented. The last mode allowed the full baseline resolution of the eight isomers on Cellulose-5 CSP, with 20% 2-PrOH in n-heptane at a flow-rate of 0.8 mL/min, at 25°C, λ = 220 nm. The limits of detection and of quantification were determined for this method and the best values obtained for isomer 8 were equal to 2.84 and 9.37 nM respectively. Finally, a small-scale preparative separation of the multiple chiral centers compound was implemented on Cellulose-5 CSP within 10% 2-PrOH in n-heptane in order to study the stereoisomer elution order on Cellulose-2, Cellulose-5 and Chiralpak AD-H, under EtOH or 2-PrOH in n-heptane mobile phases, and partial reversal elution orders were observed.

Chlorite oxidized oxyamylose differentially influences the microstructure of fibrin and self assembling peptide hydrogels as well as dental pulp stem cell behavior.

Tailored hydrogels mimicking the native extracellular environment could help overcome the high variability in outcomes within regenerative endodontics. This study aimed to evaluate the effect of the chemokine-binding and antimicrobial polymer, chlorite-oxidized oxyamylose (COAM), on the microstructural properties of fibrin and self-assembling peptide (SAP) hydrogels. A further goal was to assess the influence of the microstructural differences between the hydrogels on the in vitro behavior of human dental pulp stem cells (hDPSCs). Structural and mechanical characterization of the hydrogels with and without COAM was performed by atomic force microscopy and scanning electron microscopy to characterize their microstructure (roughness and fiber length, diameter, straightness, and alignment) and by nanoindentation to measure their stiffness (elastic modulus). Then, hDPSCs were encapsulated in hydrogels with and without COAM. Cell viability and circularity were determined using confocal microscopy, and proliferation was determined using DNA quantification. Inclusion of COAM did not alter the microstructure of the fibrin hydrogels at the fiber level while affecting the SAP hydrogel microstructure (homogeneity), leading to fiber aggregation. The stiffness of the SAP hydrogels was sevenfold higher than the fibrin hydrogels. The viability and attachment of hDPSCs were significantly higher in fibrin hydrogels than in SAP hydrogels. The DNA content was significantly affected by the hydrogel type and the presence of COAM. The microstructural stability after COAM inclusion and the favorable hDPSCs' response observed in fibrin hydrogels suggest this system as a promising carrier for COAM and application in endodontic regeneration.

How mobile phase composition and column temperature affect enantiomer elution order of liquid crystals on amylose tris(3-chloro-5-methylphenylcarbamate) as chiral selector.

A comprehensive study into the effects of mobile phase composition and column temperature on enantiomer elution order was conducted with a set of chiral rod-like liquid crystalline materials. The analytes were structurally similar and comprised variances such as length of terminal alkyl chain, presence of chlorine, number of phenyl rings, and type of chiral center. Experiments were carried out in polar organic and reversed-phase modes using amylose tris(3-chloro-5-methylphenylcarbamate) immobilized on silica gel as the chiral stationary phase. For all liquid crystals, reversal of elution order of enantiomers was observed based on type of used cosolvent and/or its content in the mobile phase; for some of the liquid crystals a temperature-induced reversal was also observed. Both linear and nonlinear dependencies of natural logarithm of enantioselectivity on temperature were found. Tested mobile phases comprised pure organic solvents and binary and tertiary mixtures of acetonitrile with organic solvents and/or water. Effect of acidic/basic mobile phase additives was also tested. Effect of structure of chiral selector is briefly discussed.

Study of enantioseparation of ß-blockers using amylose tris(3-chloro-5-methylphenylcarbamate) as chiral stationary phase under polar-organic, reversed-phase and hydrophilic interaction liquid chromatography conditions.

In this study, we describe the experimental variables influencing enantioseparation of twelve ß-blockers when analyzed under polar-organic, reversed-phase and hydrophilic interaction liquid chromatography conditions on a column with immobilized amylose tris(3-chloro-5-methylphenylcarbamate) as chiral stationary phase. Regarding polar-organic mode, two component mobile phases consisting of methanol, ethanol or acetonitrile with the addition of basic additives such as diethylamine, triethylamine, mono-ethanolamine, ethylendiamine or trifluoroacetic acid/diethylamine mixture were evaluated. Studies of retention at different temperatures were also performed. In reversed-phase mode, mixtures consisting of methanol or acetonitrile with either aqueous boric acid-borate buffer or sodium hydrogen carbonate-carbonate buffer solutions were compared aiming to study the influence of organic modifier as well as buffer type and pH on resolution. In addition, a systematic evaluation of the retention factors of ß-blockers enantiomers in hydro-organic eluents containing acetonitrile in presence of diethylamine as additive was carried out by increasing progressively the water content, in order to check for retention dependencies indicative of the interplay of both hydrophilic interaction liquid chromatography and reversed-phase modes.

Direct chromatographic methods for enantioresolution of amino acids: recent developments.

α-Amino acids are present in two opposite configurations due to the presence of a central carbon atom which is a chiral center. While L-amino acids are present in large amount in nature, only tiny quantities of their D-enantiomers exist. For a long time, D-amino acids have been considered of bacterial origin only, but recently we realized that they are present in all living organisms: notably, D-amino acids play specific and relevant functions in the different organisms. Detection and quantification of D-amino acids are mandatory to shed light on their physiological roles, especially related to foods and human health. Chromatographic techniques are among the most commonly used analytical methods for the enantioseparation of amino acids. Here, we revised the latest improvements in chromatographic direct methodologies based on chiral selectors and aimed to improve analytical speed, sensitivity, robustness, and reproducibility. While current methods are well suited for D-amino acid analysis in foodstuffs and pharmaceuticals, further improvements seem required for their simultaneous, fast and sensitive detection in biological fluids, an emerging field since D-amino acids have been proposed as biomarkers of different and relevant human pathologic states.

Utilization of the hysteresis phenomenon for chiral high-performance liquid chromatographic method selection in polar organic mode.

Polysaccharide-based chiral stationary phases (CSPs) are outstandingly suitable to play a key role in chiral HPLC method selection strategies, since they provide high success rates. One reason for this ability is that they adopt a diversity of higher order structures in various eluents, resulting in versatile chiral environments. A potential to extend this versatility further was expected and examined in the present study, based on the recently discovered hysteretic behavior of a widely used chiral selector (CS), amylose tris(3,5-dimethylphenylcarbamate). The hindered transitions of its structure, which are behind the history dependence of its separation ability, were used as a tool to identify distinct states of the chiral selector in order to exploit an extended selectivity space. The identification was carried out using a single diagnostic compound, as opposed to the common approach where testing a library of compounds is required. Eluent mixtures consisting of 2-propanol and either methanol or ethanol were scrutinized in terms of stability and robustness of the observed retentions. The solvent mixtures that were eligible for practical application in these respects were used to construct a screening sequence, including identical compositions combined with different column pretreatment. The gain achievable by using the proposed sequence was then evaluated using 15 enantiomer pairs with focus on resolution, enantiomer elution order and chemoselectivity.

Further study on enantiomer resolving ability of amylose tris(3-chloro-5-methylphenylcarbamate) covalently immobilized onto silica in nano-liquid chromatography and capillary electrochromatography.

In the present study separation of enantiomers of some chiral neutral, basic and weakly acidic analytes was investigated on the chiral stationary phase (CSP) made by covalent immobilization of amylose tris(3-chloro-5-methylphenylcarbamate) onto aminopropylsilanized (APS) silica in nano-liquid chromatography (nano-LC) in aqueous methanol or acetonitrile mixtures. It has been shown that similar to high-performance liquid chromatography (HPLC) and supercritical fluid chromatography (SFC) this chiral selector is useful for separation of enantiomers of neutral, basic and acidic analytes also in nano-LC. In comparison to our previous research, in which the chiral selector (CS) was bonded on native silica, in this study, the CS was immobilized on APS silica in order to improve chromatographic performance towards basic analytes. In fact, some improvement was observed and surprisingly not only for basic but also for neutral and acidic analytes. Again, quite unexpectedly almost no electroosmotic flow (EOF) was observed in capillaries packed with ca. 20% (w/w) amylose tris(3-chloro-5-methylphenylcarbamate) immobilized onto APS silica although the same APS silica before attachment of chiral selector exhibited significant EOF. In order to generate EOF in the capillaries with the CSP and enable capillary electrochromatographic (CEC) experiment on it, the short segment of the capillary column was packed with APS silica without chiral selector. The EOF in such capillary enabled CEC experiment and some preliminary results are reported here.

Chiral separation of several pesticides on an immobilized amylose tris(3-chloro-5-methylphenylcarbamate) column under polar-organic conditions. Influence of mobile phase and temperature on enantioselectivity.

In this work, the use of different solvents and temperatures was explored, aiming to evaluate their influence on the enantioseparation of pesticides by HPLC in polar-organic conditions, employing a column containing immobilized amylose tris(3-chloro-5-methylphenyl-carbamate). The chiral separation of seventeen different pesticides widely used as herbicides, fungicides, insecticides and precursors were studied. The mobile phases included methanol, ethanol, iso-propanol, n-propanol and acetonitrile; either pure or containing additives such as diethylamine, trifluoroacetic acid, formic acid, acetic acid or mixtures thereof. We studied the influence of these eluents on chiral separation of those pesticides in terms of retention factor, enantioselectivity, enantioresolution and peak symmetry. Regarding temperature influence, evaluated within the range 5 - 40 °C, nearly all the compounds decreased their retention and selectivity factors with the increase in temperature, although the effect was dependent on the mobile phase solvent. Moreover, estimation of thermodynamic parameters was performed based on linear van´t Hoff plots.

A Validated LC Method for the Enantiomeric Separation of EAI045 on Chiral Stationary Phase.

A simple and accurate chiral liquid chromatographic method was developed for enantiomeric resolution and determination of 2-(5-fluoro-2-hydroxyphenyl)-2-(1-oxo-2,3-dihydro-1H-isoindol-2-yl)-N-(1,3-thiazol-2-yl)acetamide (EAI045). The enantiomers of EAI045 were baseline resolved on a Chiralpak AD-H (250 mm × 4.6 mm, 5 µm) column using a mobile phase system containing n-hexane: 2-propanol (75: 25 v/v) at a flow rate of 1 mL min-1 at 30°C. The eluted analytes were subsequently detected with an ultraviolet detector at 254 nm. The effects of organic modifiers and temperature on the enantioselectivity and resolution of the enantiomers were evaluated. The calibration curves were plotted within the concentration range between 2 and 600 µg mL-1 (n = 11), and recoveries between 98.74% and 101.52% were obtained, with relative standard deviation < 1.4%. The limit of detection and limit of quantitation for R-enantiomer were 0.94 and 3.07 µg mL-1 and for S-enantiomer were 0.86 and 2.84 µg mL-1, respectively. The validated method was found to be suitable for enantiomeric separation and sufficiently accurate for the determination of enantiomeric purity of EAI045 in bulk drugs.

Comparison of dimethylated and methylchlorinated amylose stationary phases, coated and covalently immobilized on silica, for the separation of some chiral compounds in supercritical fluid chromatography.

The chromatographic properties of a new coated amylose tris(3-chloro-5-methylphenylcarbamate) were evaluated in supercritical fluid chromatography for the separation of enantiomers of chiral 1-aryl-5-aryl-pyrrolidin-2-one derivatives, potential anticancer agents, and some commercial drugs. The mobile phase consisted of CO2-modifier mixtures with 30% of either methanol or ethanol, the flow rate was 3 mL/min. The column oven temperature was 40 °C and the outlet pressure was 15 MPa, in order to limit the compressibility of the CO2, thus limiting density variation along the column. The obtained results were then compared to those observed toward 3 other stationary phases: the coated amylose tris(3,5-dimethylphenylcarbamate), the immobilized amylose tris(3,5-dimethylphenylcarbamate) and the coated amylose tris(5-chloro-2-methylphenylcarbamate). It was shown that the new coated amylose tris(3-chloro-5-methylphenylcarbamate) was the most retentive column whatever the studied compounds, particularly for thalidomide and omeprazole with retention factors up to 73.3 and 29.5for the second enantiomer, respectively. Concerning the enantioselectivity, even most of the compounds are separated on all the four columns, the coated amylose tris(3-chloro-5-methylphenylcarbamate) allows the best resolution for most of the ten studied analytes (except omeprazole for which the resolution values are equal to 7.8 and 9.7 on the coated amylose tris(3-chloro-5-methylphenylcarbamate) and amylose tris(3,5-dimethylphenylcarbamate), respectively). Acting in complementary ways, the two chlorinated stationary phases permitted the complete separation of enantiomers of nine compounds out of the ten.

High-performance liquid chromatographic enantioseparation of isopulegol-based ß-amino lactone and ß-amino amide analogs on polysaccharide-based chiral stationary phases focusing on the change of the enantiomer elution order.

The enantioselective separation of newly prepared, pharmacologically significant isopulegol-based ß-amino lactones and ß-amino amides has been studied by carrying out high-performance liquid chromatography on diverse amylose and cellulose tris-(phenylcarbamate)-based chiral stationary phases (CSPs) in n-hexane/alcohol/diethylamine or n-heptane/alcohol/ diethylamine mobile phase systems. For the elucidation of mechanistic details of the chiral recognition, seven polysaccharide-based CSPs were employed under normal-phase conditions. The effect of the nature of selector backbone (amylose or cellulose) and the position of substituents of the tris-(phenylcarbamate) moiety was evaluated. Due to the complex structure and solvation state of polysaccharide-based selectors and the resulting enantioselective interaction sites, the chromatographic conditions (e.g., the nature and content of alcohol modifier) were found to exert a strong influence on the chiral recognition process, resulting in a particular elution order of the resolved enantiomers. Since no prediction can be made for the observed enantiomeric resolution, special attention has been paid to the identification of the elution sequences. The comparison between the effectiveness of covalently immobilized and coated polysaccharide phases allows the conclusion that, in several cases, the application of coated phases can be more advantageous. However, in general, the immobilized phases may be preferred due to their increased robustness. Thermodynamic parameters derived from the temperature-dependence of the selectivity revealed enthalpically-driven separations in most cases, but unusual temperature behavior was also observed.