Cationic cyclodextrins chemically-bonded chiral stationary phases for high-performance liquid chromatography.

Two covalently bonded cationic ß-CD chiral stationary phases (CSPs) prepared by graft polymerization of 6(A)-(3-vinylimidazolium)-6-deoxyperphenylcarbamate-ß-cyclodextrin chloride or 6(A)-(N,N-allylmethylammonium)-6-deoxyperphenylcarbamoyl-ß-cyclodextrin chloride onto silica gel were successfully applied in high-performance liquid chromatography (HPLC). Their enantioseparation capability was examined with 12 racemic pharmaceuticals and 6 carboxylic acids. The results indicated that imidazolium-containing ß-CD CSP afforded more favorable enantioseparations than that containing ammonium moiety under normal-phase HPLC. The cationic moiety on ß-CD CSPs could form strong hydrogen bonding with analytes in normal-phase liquid chromatography (NPLC) to enhance the analytes' retention and enantioseparations. In reversed-phase liquid chromatography (RPLC), the analytes exhibited their maximum retention when the pH of mobile phase was close to their pK(a) value. Inclusion complexation with CD cavity and columbic/ionic interactions with cationic substituent on the CD rim would afford accentuated retention and enantioseparations of the analytes.

Chemically bonded cationic ß-cyclodextrin derivatives and their applications in supercritical fluid chromatography.

Cationic ß-cyclodextrin (CD) perphenylcarbamoylated derivatives were chemically bonded onto vinylized silica using a radical co-polymerization reaction. The derived materials were used as chiral stationary phases (CSP) in supercritical fluid chromatography (SFC). Enantioseparations were successfully demonstrated on 14 racemates encompassing flavanones, thiazides and amino acid derivatives. The electrostatic force between the analytes and the cationic moiety on ß-CD derivative was found to be important for retention and enantioseparation of the racemates. Aromatic cationic moiety on ß-CD enabled better enantioseparations than aliphatic cationic moiety. It was also found that the presence of acid additives would result in lower retention of the analytes but often assist the chiral resolutions.

Chiral capillary electrophoresis with cationic pyrrolidinium-beta-cyclodextrin derivatives as chiral selectors.

New single-isomer, cationic beta-cyclodextrins, including mono-6-deoxy-6-pyrrolidine-beta-cyclodextrin chloride (pyCDCl), mono-6-deoxy-6-(N-methyl-pyrrolidine)-beta-cyclodextrin chloride (N-CH(3)-pyCDCl), mono-6-deoxy-6-(N-(2-hydroxyethyl)-pyrrolidine)-beta-cyclodextrin chloride (N-EtOH-pyCDCl), mono-6-deoxy-6-(2-hydroxymethyl-pyrrolidine)-beta-cyclodextrin chloride (2-MeOH-pyCDCl) were synthesized and used as chiral selectors in capillary electrophoresis for the enantioseparation of carboxylic and hydroxycarboxylic acids and dansyl amino acids. The unsubstituted pyCDCl exhibited the greatest resolving ability. Most analytes were resolved over a wide range of pH from 6.0 to 9.0 with this chiral selector. In general, increasing pH led to a decrease in resolution. The effective mobilities of all the analytes were found to decrease with increasing CD concentration. The optimal concentration for most carboxylic acids and dansyl amino acid was in the range 5-7.5 mM and >15 mM for hydroxycarboxylic acids. (1)H NMR experiments provided direct evidence of inclusion in the CD cavity.

Synthesis of large pore-diameter SBA-15 mesostructured spherical silica and its application in ultra-high-performance liquid chromatography.

A modified synthesis approach for spherical large pore-diameter SBA-15 mesoporous silica (SLP-SBA-15) with particle size range of 0.5-1microm was being reported. It was worth mentioning that in this improved methodology, the use of cetyltrimethylammonium bromide (CTAB) as co-template significantly reduced the self-assembly time from 24h to 45min. Moreover, under reflux condition, the reaction time could be further shortened by reducing the aging time from 48h to 6h. The resultant SLP-SBA-15 was thereafter successfully functionalized and packed into an ultra-high-performance liquid chromatography (UHPLC) column for the separation of aromatic compounds. A variety of characterizations demonstrated that the silica products exhibited a well-ordered 2d hexagonal mesostructure with well-formed spherical morphology. pore-diameter can be enlarged up to 8.2nm without affecting the structural order. The SLP-SBA-15 samples showed excellent thermal and hydrothermal stabilities. The octadecyltriethoxysilane functionalized SLP-SBA-15 (SLP-C18-SBA-15) was demonstrated to be an effective stationary phase in UHPLC application because the column exhibited significantly reduced column pressure (2800psi) at a flow rate of 0.4ml/min. Accordingly, it would afford greater flexibility for tuning of the flow rate to meet the fast separation requirement.

Enantioseparation of a novel "click" chemistry derived native beta-cyclodextrin chiral stationary phase for high-performance liquid chromatography.

A novel native beta-cyclodextrin chiral stationary phase was prepared via "click" chemistry with cuprous iodide-triphenylphosphine complex as the catalyst and applied for enantioseparation of Dns-amino acids, substituted phenyl and phenoxy group modified propionic acids, flavonoids, and some pharmaceutical compounds such as nimodipine, propranolol, brompheniramine and bendroflumethiazide in reversed-phase high-performance liquid chromatography. The studied analytes could be resolved under different separation conditions. The resolution of Dns-DL-Leu could reach 5.08 using a mobile phase consisting of 1% (w/w) triethylammonium acetate buffer (pH 4.11) and methanol (50:50 v/v). The effects of buffer pH and the content of organic modifier on enantioseparation of Dns-amino acids by this novel chiral phase were being investigated. The separation results demonstrate that click chemistry, a versatile reaction, affords a facile approach towards the preparation of stable chiral stationary phases.

Synthesis and application of a novel single-isomer mono-6-deoxy-6-(3R,4R-dihydroxypyrrolidine)-beta-cyclodextrin chloride as a chiral selector in capillary electrophoresis.

A novel positively charged single-isomer of beta-cyclodextrin, mono-6-deoxy-6-(3R,4R-dihydroxypyrrolidine)-beta-CD chloride (dhypy-CDCl), was synthesized and employed as a chiral selector for the first time in capillary electrophoresis (CE) for the enantioseparation of anionic and ampholytic acids. The effects of the running buffer pH, chiral selector concentration, analyte structure and organic modifier on the enantioseparation were studied in detail. The chiral selectivity and resolution for most of the studied analytes decreased as the buffer pH increased in the range of 6.0-9.0. Increasing selector concentration led to decreased effective mobility, increased chiral selectivity and resolution for most of the studied analytes. Moreover, the hydroxyl groups located on the dihydroxypyrrolidine substituent of the dhypy-CDCl could have influence on the chiral separation.

Synthesis of cationic beta-cyclodextrin derivatives and their applications as chiral stationary phases for high-performance liquid chromatography and supercritical fluid chromatography.

Four cationic beta-cyclodextrin derivatives, namely mono-6-(3-methylimidazolium)-6-deoxy-perphenylcarbamoyl-beta-cyclodextrin chloride (MPCCD), mono-6-(3-methylimidazolium)-6-deoxyper(3,5-dimethylphenylcarbamoyl)-beta-cyclodextrin chloride (MDPCCD), mono-6-(3-octylimidazolium)-6-deoxyperphenylcarbamoyl-beta-cyclodextrin chloride (OPCCD) and mono-6-(3-octylimidazolium)-6-deoxyper(3,5-dimethylphenylcarbamoyl)-beta-cyclodextrin chloride (ODPCCD), have been synthesized and physically coated onto porous spherical silica gel to obtain novel chiral stationary phases (CSPs). The performances of these CSPs are studied on high-performance liquid chromatography (HPLC) and supercritical fluid chromatography (SFC) using 18 racemic aryl alcohols as test analytes. Among these four CSPs, OPCCD shows the best separation results for all analytes on both HPLC and SFC analyses. Chromatographic studies reveal that the CSPs consisting of an n-octyl group on the imidazolium moiety and phenylcarbamoyl groups on the cyclodextrin ring provide enhancement of analyte-chiral substrate interactions over CSPs bearing the methyl group on the imidazolium moiety and 3,5-dimethylphenylcarbamoyl groups on the cyclodextrin ring.

Synthesis and application of mono-6-(3-methylimidazolium)-6-deoxyperphenylcarbamoyl-beta-cyclodextrin chloride as chiral stationary phases for high-performance liquid chromatography and supercritical fluid chromatography.

The synthesis of mono-6-(3-methylimidazolium)-6-deoxyperphenylcarbamoyl-beta-cyclodextrin chloride (MPCCD) and its application in chiral stationary phases (CSPs) for high-performance liquid chromatography (HPLC) and supercritical fluid chromatography (SFC) are being reported. This chiral selector is coated onto silica gel in different weight percentages (15, 20 and 35%, w/w) to obtain CSPs having different loading content. These new chiral stationary phases are tested using normal-phase HPLC for enantioseparation of racemic aromatic alcohols. Indeed, the enantiodiscrimination abilities of these CSPs are found to be influenced by the loading content of the chiral selector. Among the three columns (MPCCD-C15, MPCCD-C20 and MPCCD-C35), the best enantioseparation results are obtained using a column containing 20% (w/w) of MPCCD (MPCCD-C20). The resolution (R(s)) obtained for p-fluorophenylethanol, p-chlorophenylethanol, p-bromophenylethanol, p-iodophenylethanol and p-fluorophenyl-3-buten-1-ol using MPCCD-C20 ranges from 3.83 to 5.65. Good enantioseparation results are obtained for these analytes under SFC separation conditions using the MPCCD-C20 column.

Chiral separation of dansyl amino acids in capillary electrophoresis using mono-(3-methyl-imidazolium)-beta-cyclodextrin chloride as selector.

Enantioseparations of fourteen dansyl amino acids were achieved by using a positively-charged single-isomer beta-cyclodextrin, mono-(3-methyl-imidazolium)-beta-cyclodextrin chloride, as a chiral selector. Separation parameters such as buffer pH, selector concentration, separation temperature, and organic modifier were investigated for the enantioseparation in order to achieve the maximum possible resolution. Chiral separation of dansyl amino acids was found to be highly dependent on pH since the degree of protonation of these amino acids can alter the strength of electrostatic interaction and/or inclusion complexation between each enantiomer and chiral selector. In general, the chiral resolution of dansyl amino acids was enhanced at higher pH, which indicates that the carboxylate group on the analytes may interact with the imidazolium group of cationic cyclodextrin. For most analytes, a distinct maximum in enantioresolution was obtained at pH 8.0. Moreover, the chiral separation can be further improved by careful tuning of the separation parameters such as higher selector concentration (e.g. 10 mM), lower temperature, and addition of methanol. Enantioseparation of a standard mixture of these dansyl amino acids was further achieved in a single run within 30 min.

Effect of alkylimidazolium substituents on enantioseparation ability of single-isomer alkylimidazolium-beta-cyclodextrin derivatives in capillary electrophoresis.

A family of 6-mono(3-alkylimidazolium)-beta-cyclodextrins with one primary hydroxyl group replaced by an alkylimidazolium cation has been developed. The effect of alkyl substitutents on the enantioresolution ability of these single-isomer cyclodextrins towards dansyl amino acids has been studied by capillary electrophoresis. Systematical investigations on the effect of buffer pH and selector concentration on the enatioseparation show that chiral selectors with a shorter alkyl chain (R=C(n)H(2n+1), n

Urea bonded cyclodextrin derivatives onto silica for chiral HPLC.

Several structurally well-defined perfunctionalised cyclodextrin chiral stationary phases (CD CSPs) for high performance liquid chromatography have been successfully prepared by immobilisation of perfunctionalised cyclodextrins on silica through urea linkage(s) using the Staudinger reaction. These CSPs show high chiral recognition efficiency and are utilised in the resolution of various types of racemic compounds. This paper reviews the development of sixteen perfunctionalised cyclodextrin-based CSPs, their preparation, and their application to enantioseparation of seventy-seven racemic compounds under a range of separation conditions.

Synthesis and application of single-isomer 6-mono(alkylimidazolium)-beta-cyclodextrins as chiral selectors in chiral capillary electrophoresis.

Novel single isomers of positively charged beta-CDs were prepared via nucleophilic substitution of 6-monotosyl-beta-CD with alkylimidazoles to afford 6-mono(alkylimidazolium)-beta-CD tosylates and then 6-mono(alkylimidazolium)-beta-CD chlorides by anion exchange. The chiral resolution abilities of these cationic CDs were studied by CE using dansyl (Dns)-amino acids as model analytes. From the experimental results, it was found that both resolution and selectivity of these cationic CDs were dependent on the following parameters: concentration of chiral selectors, pH of the running buffer, counteranions of the CDs, side chain length of the n-alkyl-imidazolium cation, temperature of the capillary column, and organic modifier used. The concentration of chiral selectors required for enantioseparation varied from 3 to 30 mM. The BGE pH played an important role in the resolution of Dns-amino acids. For acidic BGEs, chiral resolution increased with pH (4.0-6.0) and reached a local maximum at pH 6.0. However, better resolutions were obtained with basic phosphate buffer at pH 9.6. Methanol was found to be an effective organic modifier for the resolution of Dns-amino acids by CE.

A family of single-isomer positively charged cyclodextrins as chiral selectors for capillary electrophoresis: mono-6A-butylammonium-6A-deoxy-beta-cyclodextrin tosylate.

A novel single-isomer positively charged beta-cyclodextrin (beta-CD), mono-6(A)-butylammonium-6(A)-deoxy-beta-cyclodextrin tosylate (BuAM-beta-CD), has been synthesized, characterized, and used for the enantioseparations of alpha-hydroxy acids, carboxylic acids, and ampholytic analytes by capillary electrophoresis in acidic aqueous background electrolytes. The effective mobilities of all studied analytes decreased with increasing concentration of CD. Satisfactory resolutions were obtained for alpha-hydroxy acids over a wide range of chiral selector concentration. The optimum CD concentration was lower than 5 mM for the carboxylic acids, while higher than 20 mM for alpha-hydroxy acids. Inclusion complexation in combination with ion pair interaction seemed to account for the chiral discrimination process. The hydrogen bonding may provide secondary contribution for the chiral resolution of alpha-hydroxy acids. In addition, BuAM-beta-CD was further proved to be an effective chiral selector for anionic analytes by the baseline enantioseparation of a six-acid mixture within 20 min.

Enantioseparation of dansyl amino acids by a novel permanently positively charged single-isomer cyclodextrin: Mono-6-N-allylammonium-6-deoxy-ß-cyclodextrin chloride by capillary electrophoresis.

A permanently positively charged single-isomer ß-CD, mono-6-N-allylammonium-6-deoxy-ß-CD chloride, ALAM-ß-CD, has been synthesized and successfully used as chiral selector in capillary electrophoresis for enantioseparation of amino acids and dansyl amino acids. The effects of pH, CD's concentration and capillary length on enantioseparation were studied in order to investigate its potential as chiral selector for acidic racemates. By increasing buffer's pH and CD's concentration, fairly robust separations were achieved. Good separations were also obtained with a shorter capillary. In addition, baseline enantiomeric separations of a mixture of six pairs of dansyl dl-amino acids were achieved under different pH conditions.

Enantioseparation of acidic enantiomers in capillary electrophoresis using a novel single-isomer of positively charged beta-cyclodextrin: Mono-6A-N-pentylammonium-6A-deoxy- beta-cyclodextrin chloride.

The new single-isomer of positively charged beta-cyclodextrin, mono-6A-N-pentylammonium-6A-deoxy-beta-cyclodextrin chloride (PeAM-beta-CD), was employed for the first time for the enantioseparation of anionic and ampholytic analytes by capillary electrophoresis (CE). The synthesis and characterization of PeAM-beta-CD were reported. The effect of background electrolyte (BGE) pH and selector concentration on the enantioseparation was investigated. Good separation was obtained at low BGE pH (ca. 5.0-6.0). The effective mobilities of all analytes were found to decrease with increasing CD concentration. PeAM-beta-CD proved to be an effective chiral selector for most studied anionic analytes.

Enantiomer separation of flavour and fragrance compounds by liquid chromatography using novel urea-covalent bonded methylated beta-cyclodextrins on silica.

A novel methylated beta-cyclodextrin chiral stationary phase (CSP-ME), which was chemically immobilised onto porous silica via multiple urea-linkages was synthesised. The CSP-ME chiral stationary phase depicted good enantiomer separation abilities for some well-known flavour as well as fragrance compounds using high-performance liquid chromatography under reverse phase conditions. The optimum resolution for alpha-ionone, 3-methyl-alpha-ionone, flavanone, 5-methoxyflavanone, 6-methoxyflavanone, 7-methoxyflavanone, hesperetin, naringenin and taxifolin was achieved using a mobile phase composition consisting of 1 wt.% triethylammonium acetate buffer (pH 4.68)-methanol. The effects of pH of triethylammonium acetate buffer and the methanol-acetonitrile content of the mobile phase composition on their retention time and resolution were examined to optimise the separation conditions.