Extending the scope of enantiomer separation by capillary supercritical fluid chromatography on immobilized polysiloxane-anchored permethyl-β-cyclodextrin (Chirasil-Dex)

Immobilized polysiloxane-anchored permethyl-β-cyclodextrin (Chirasil-Dex) with a cyclodextrin content of approximately 30 % by weight, previously employed as a versatile chiral stationary phase for the separation of enantiomers by GC, has been used for the separation of enantiomers by capillary supercritical fluid chromatography (SFC). A considerable number of racemates could be resolved, e.g. aromatic alcohols, amino alcohols (TFA derivatives), and underivatized acids. Many pharmaceutical compounds were among those analyzed, including several NSAIDs (e.g. ibuprofen and ketoprofen), a steroidal drug (nor-gestrel), a barbiturate (hexobarbital), and others. Among the racemates resolved were many which cannot be analyzed by GC owing to low volatility or decomposition at elevated temperatures. For two racemates, analysis temperature and mobile phase density were systematically varied to give constant analysis times or capacity factors k. Low temperatures (ca 60 °C) yielded the best separation in term of separation factor, α, or resolution, R_s, even though higher densities had to be used. In comparison with GC, capillary SFC was able to furnish higher separation factors and similar resolution. The applicability of capillary SFC for the analysis of mixtures of cyclodextrin derivatives, e.g. those used in the synthesis of Chirasil-Dex, was, furthermore, demonstrated.     

Unusual peak defocusing in capillary GC on hexakis(2,6-diO-pentyl-3-O-acetyl)-α-cyclodextrin stationary phase

An unusual peak defocusing effect influencing chromatographic performance over a limited range of elution temperatures is described for hexakis(2,6-di-O-pentyl-3-O-acetyl)-α-cyclodextrin stationary phase. Since this phenomenon is likely to be dependent on minor details of the cyclodextrin molecule, full assignment of the ¹H- and ¹³C-NMR-spectra are given.     

Cyclodextrins as chiral stationary phases in capillary gas chromatography. Part III: Hexakis(3-O-acetyl-2,6-di-O-pentyl)-α-cyclodextrin

The properties of hexakis(3-O-acetyl-2,6-di-O-pentyl)-α-cyclodextrin as a chiral stationary phase for capillary gas chromatography are described. For the first time the enantiomers of a series of different lactones are separated and their order of elution is assigned. Moreover, the enantiomers of trifluoroacetylated aldols and amino alcohols, the cyclic carbonates of 1,2-diols, 1,3-diols, O-alkylated glycerols, and some chiral pharmaceuticals are also separated on the new chiral phase. The modified α-cyclodextrin is stable above 200°C.     

Enantiomer separation of side-chain fluorinated alkylbenzenes by capillary gas chromatography on cyclodextrin phases

Summary The separation of racemic side-chain fluorinated alkylbenzenes and bromofluorinated analogues by capillary gas chromatography using permethylated , and -cyclodextrins dissolved in polysiloxanes of different polarity as stationary phases is described. The influence of the achiral polysiloxane matrices on the separation of enantiomers is discussed in the light of the results obtained with the different phases. For a part of the tested compounds thermodynamic data are determined which describe the interaction of enantiomers with the stationary phase. The mechanism of separation is discussed on this basis and by comparison with data for structurally similar compounds.     

Cyclodextrin derivatives for the GC separation of racemic mixtures of volatile compounds. Part VI: The influence of the diluting phase on the enantioselectivity of 2,6-Di-O-methyl-3-O-pentyl-β-cyclodextrin

As a continuation of previous studies on the use of cyclodextrin derivatives (CD) for the separation of volatile compounds by capillary GC, the influence of diluting phases other than OV-1701 or OV-1701-OH has been investigated. 2,6-Di-O-methyl-3-O-pentyl-β-cyclodextrin (2,6-DiMe-3-Pe-β-CD) was taken as the reference CD derivative, because of the large number of volatile racemates it is able to separate; OV-1701 or OV-1701-OH was chosen as the reference diluting phase. The performance of a column coated with a 0.15 μm film of 10 % 2,6-DiMe-3-Pe-β-CD in OV-1701 was compared with that of analogous columns coated with films of the same thickness containing the same percentage of the CD derivative diluted with stationary phases of different polarity, i.e. PS-086, PS-347.5, and OV-225. Resolution values and separation factors of thirty racemates were used to evaluate the effect of different diluting phases on column performance.     

Cyclodextrins as chiral stationary phases in capillary gas chromatography. Part V: Octakis(3-O-butyryl-2,6-di-O-pentyl)-γ-cyclodextrin

γ-Cyclodextrin with 3-O-butyryl and 2,6-di-O-pentyl residues is a very versatile chiral stationary phase for enantiomer separation. Most of the common and many uncommon amino acids can be separated as well as α- and β-hydroxy acids, chiral alcohols, diols, triols, ketones, bicyclic, and tricyclic acetals, amines, alkyl halides, lactones, and functionalized cyclopropane derivatives.     

Cyclodextrin derivatives for GC separation of racemic mixtures of volatile compounds. Part VIII: 2,6-di-O-methyl-3-O-pentyl-γ-cyclodextrin and 2,6-di-O-methyl-3-O-(4-oxo-pentyl)- and 2,6-di-O-pentyl-3-O-(4-oxo-pentyl)-β-and -γ-cyclodextrins

In pre vious papers, 2,6-di-O-methyl-3-O-pentyl-β-cyclodextrin (CD) was demonstrated to be successful in separating volatile compounds, while avoiding the drawbacks of 2,3,6-tri-O-methyl-O-methyl-β-CD in terms of column stability and operating temperature. Since a CD chiral selector of universal use has not yet been found, and at least two (or more) columns coated with different CD derivatives are therefore necessary for routine work, the performance of 2,6-di-O-methyl-3-O-pentyl-γ-CD, 2,6-di-O-methyl-3-O-(4-oxopentyl)-γ-CD, 2,6-di-O-pentyl-3-O-(4-oxo-pentyl)-β-CD, and 2,6-di-O-pentyl-3-O-(-4-oxo-pentyl)-γ-CD diluted in polysiloxanes for the separation of volatile compounds in aromas and essential oils will be illustrated; each column coated with each of the newly synthesized CD derivatives was evaluated by analyzing more than 150 different recemates with different structures.     

Enantiomeric separation of six chiral pesticides that contain chiral sulfur/phosphorus atoms by supercritical fluid chromatography

Six chiral pesticides containing chiral sulfur/phosphorus atoms were separated by supercritical fluid chromatography with supercritical CO₂ as the main mobile phase component. The effect of the chiral stationary phase, different type and concentration of modifiers, column temperature, and backpressure on the separation efficiency was investigated to obtain the appropriate separation condition. Five chiral pesticides (isofenphos‐methyl, isocarbophos, flufiprole, fipronil, and ethiprole) were baseline separated under experimental conditions, while isofenphos only obtained partial separation. The Chiralpak AD‐3 column showed a better chiral separation ability than others for chiral pesticides containing chiral sulfur/phosphorus atoms. When different modifiers at the same concentration were used, the retention factor of pesticides except flufiprole decreased in the order of isopropanol, ethanol, methanol; meanwhile, the retention factor of flufiprole increased in the order of isopropanol, ethanol, methanol. For a given modifier, the retention factor and resolution decreased on the whole with the increase of its concentration. The enantiomer separation of five chiral pesticides was an “enthalpy‐driven” process, and the separation factor decreased as the temperature increased. The backpressure of the mobile phase had little effect on the separation factor and resolution.     

Enantiomer separation by gas chromatography on cyclodextrin chiral stationary phases

Fused silica capillary columns coated with several alkyl or acyl cyclodextrin derivatives, especially those of α- and β-cyclodextrins, are suitable for the enantiomer separation of a wide variety of volatile compounds of different molecular size and functionality. Positional isomers and more than 250 pairs of optical isomers have been resolved, including chiral hydrocarbons, acetals, ethers, epoxides, carbonates, lactones, esters, acids, ketones, aldehydes, alcohols, halocarbons, and also nitrogen-and sulfur-containing compounds. The physical properties of the cyclodextrin derivatives, even those obtained as viscous fluids, could be improved by dissolving them in polysiloxane liquid phases commonly used for GLC.     

Preparation and characterization of phenyl carbamate derivative β-cyclodextrin bonded phase for chiral HPLC

A partially substituted β-cyclodextrin chiral stationary phase was prepared by the reaction of phenyl isocyanate. The enantiomers of a series of O,O-diethyl(p-methylbenzenesulfonamido)-aryl(or alkyl)-methylphosphonates were studied on the prepared phenyl carbamate derivative β-cyclodextrin bonded phase and a commercial (S)-(+)-1–(1-naphthyl) ethylcarbamate derivative β-cyclodextrin bonded phase on normal phase chromatographic condition. Results show that the prepared phenyl carbamate derivative β-cyclodextrin bonded phase has better enantiomeric selectivity to the series of compounds. A chiral recognition mechanism was suggested for the separation of these novel organic phosphorus enantiomers.     

Chirasil-Dex-TFA: A new polysiloxane-bonded and immobilizable cyclodextrin stationary phase for enantiomer separation by GC

Heptakis(2,6-di-O-methyl-3-O-trifluoroacetyl)-?-cyclodextrin has been chemically bonded to a polydimethylsiloxane via an octamethylene spacer and subsequently immobilized on to the surface of fused silica capillaries, the first time a cyclodextrin derivative with mixed substituents has been immobilized in this way. The enantioselectivity of the new chiral stationary phase has been investigated as a function of the cyclodextrin content. It was found to be higher than that reported for heptakis (2,6-di-O-methyl-3-O-trifluoroacetyl)-?-cyclodextrin or hepta-kis(2,6-di-O-methyl-3-O-acetyl)-?-cyclodextrin dissolved in polysiloxane (e.g. OV-1701). The degree of immobilization was over 90 %. The temperature stability is, however, inferior to that of Chirasil-Dex (a permethylated cyclodextrin): the maximum operating temperature is approximately 200 °C.     

Cyclodextrins as chiral stationary phases in capillary gas chromatography. Part VI: Octakis (2,3,6-tri-O-pentyl)-γ-cyclodextrin

Perpentylated γ-cyclodextrin is a very hydrophobic liquid which exhibits enantioselectivity toward a number of non-polar chiral substrates including the saturated hydrocarbons cis- and trans-pinane, β-chiral olefins, iron(O) tricarbonyl complexes of prochiral olefins and 3,3,8,8-tetramethyl-trans-cyclooctene, a compound with planar chirality.     

Cyclodextrins as chiral stationary phases in capillary gas chromatography. Part IV: Heptakis(2,3,6-tri-O-pentyl)-β-cyclodextrin

O-Perpentylated β-cyclodextrin has been evaluated as chiral stationary phase in capillary gas chromatography. Enantioselectivity is observed towards many chiral hydroxy compounds, including cyanohydrins and carbohydrates. Most importantly, the enantiomers of many olefins and alkyl halides can be resolved on this chiral phase. The thermal stability of the cyclodextrin derivative exceeds 200°C.     

Fast separation of flavonoids by supercritical fluid chromatography using a column packed with a sub‐2 μm particle stationary phase

The purpose of this study was to compare the effects of different chromatographic columns for the separation of seven flavonoids. Four different stationary phases are available, including bridged ethyl hybrid, BEH and the same hybrid phase modified with 2‐ethylpyridine, CSH fluorophenyl, and HSS C₁₈ SB. The analytes included calycosin, genistein, medicarpin, calycosin‐7‐O‐β‐d ‐glucoside, formononetin, formononetin‐7‐O‐β‐d ‐glucoside, and liquiritigenin. The CSH fluorophenyl column was determined to be the most suitable and provided the fastest separation within 17 min using gradient elution with carbon dioxide as the mobile phase and methanol as the co‐solvent. Good peak shapes were obtained, and the values of the peak asymmetry were close to 1.0 for all of the flavonoids. The resolution was more than 1.41 for all of the separated peaks. Baseline separation on the optimal columns was achieved by changing the co‐solvent type and adjusting the temperature and pressure. Quantitative performance was evaluated under optimized conditions, and method validation was accomplished. The validation parameters, such as linearity, sensitivity, precision, and accuracy, were satisfactory. Good repeatability of both peak area (relative standard deviation <1.02%) and retention time (relative standard deviation <0.88%) was observed. The optimized chromatographic methods were successfully used for the determination of seven flavonoids in Radix astragali . The sensitivity was sufficient for the analysis of real samples.     

Mono(6-N-allylamino-6-deoxy)perphenylcarbamoylated β-cyclodextrin: synthesis and application as a chiral stationary phase for HPLC

A novel cyclodextrin derivative: mono(6^A-N-allylamino-6^A-deoxy)perphenylcarbamoylated β-cyclodextrin was synthesized. Hydrosilylation with (EtO)₃SiH and then reaction of the reactive siloxane with pristine silica gel afforded a facile entry into a durable, structurally well-defined chiral stationary phase capable of enantioseparation of a variety of racemic drugs.     

Thiol‐ene click derived structurally well‐defined per(3,5‐dimethyl)phenylcarbamoylated cationic cyclodextrin separation material for achiral and chiral chromatography

In this work, a novel single sulfoether‐bridged cationic per(3,5‐dimethyl)phenylcarbamoylated‐β‐cyclodextrin separation material was prepared by thiol‐ene click chemistry and characterized by using FTIR spectroscopy, solid‐state ¹³C NMR spectroscopy and elemental analysis, which confirmed the correct structure. The separation material exhibited a good achiral separation performance for benzene homologues and phenylamine analogs, especially o‐xylene and m‐xylene, and m‐phenylenediamine and o‐phenylenediamine can be discriminated by the (3,5‐dimethyl)phenylcarbamoyl cyclodextrins. The chiral resolving ability of the separation material was evaluated by discriminating various isoxazolines, flavonoids, and β‐blockers in reversed‐phase high‐performance liquid chromatography. For isoxazolines, the material showed the best chiral discrimination toward 3‐aryl‐5‐(2‐oxopyrrolidin‐1‐yl)‐isoxazolines, where the resolution for 3ClPh‐OPr  reached 6.03. For flavonoids, it exhibited more efficient separation to the ones with more hydrophobic substituents, with a resolution of 5.93 for 6‐hydroxyflavanone. β‐Blockers were also enantioseparated satisfactorily on the material. The as‐prepared separation material is a good member of the thiol‐ene click derived cyclodextrin stationary phase family.