Carbamoylated azithromycin incorporated zirconia hybrid monolith for enantioseparation of acidic chiral drugs using non-aqueous capillary electrochromatography.

Carbamoylated derivatives of two antibiotics, namely, clindamycin phosphate (CLIP) and erythromycin (ERY) were successfully employed as co-precursors, in combination of zirconium tetrabutoxide as a precursor, to prepare chiral organic-zirconia hybrid monoliths (i.e., CLIP-ZHMs and ERY-ZHMs, respectively) via a single-step in-situ sol-gel approach in our previous works. Their superiority over chiral organic-zirconia/silica monoliths, prepared by post-modification approach, in terms of better enantioresolution and enhanced stability inspired us to prepare ZHMs based on an another antibiotic, azithromycin (i.e., AZI-ZHMs). Monolithic columns were employed for capillary electrochromatographic enantioseparation of acidic chiral drugs in mobile phases consisting of acetonitrile (ACN) and methanol (MeOH) as organic modifiers, and acetic acid (AcOH) and triethylamine (TEA) as electrolytes. The effects of composition of mobile phase and applied voltage on chiral separation were investigated by using ketoprofen as a representative analyte. Baseline resolutions were obtained for six acidic drugs in mobile phase consisting of 80/20 (v/v) ACN/MeOH with 300mM AcOH and 10mM TEA at a 10kV applied voltage and 25°C capillary temperature. The relative standard deviations for resolution values regarding column to column and batch to batch repeatability were less than 2.5% (for n=3) under optimized conditions, indicating satisfactory stability of the columns and reproducibility of the column preparation process.

Doxycycline as a new chiral selector in capillary electrophoresis.

Doxycycline (DOX) is one of the tetracycline class of antibiotics and has not been examined for its enantioseparation abilities previously. The purpose of the present work was to evaluate DOX as a chiral selector (CS) using capillary electrophoresis (CE) in nonaqueous mode. Systematic experiments were performed to investigate the effects of concentration of CS, compositions of organic solvents and background electrolytes, applied voltage on chiral separation of a set of acidic chiral compounds. Excellent resolutions of enantiomers of acidic chiral compounds were attained in a background electrolytes composed of 50:50 acetonitrile/methanol+214mM acetic acid+ 63mM triethylamine and 38mM DOX. The results show that DOX is a viable chiral selector in CE for the enantioseparation of the type of chiral compounds investigated.

Application of rifampicin as a chiral selector for enantioresolution of basic drugs using capillary electrophoresis.

Rifampicin, a member of rifamycin sub-class of antibiotics which belongs to the naphthalenic ansamycin class of antibiotics, has a characteristic ansa structure, i.e., a ring structure or chromophore spanned by an aliphatic chain. The present work was designed to evaluate its potential as a chiral selector (CS) as its structure consisting of nine stereogenic centers, an aromatic moiety and several functional groups (i.e., one imine, one amide, one acetoxy residue, two aliphatic hydroxyl and three phenolic hydroxyl groups) was expected to instigate multiple enantioselective interactions, namely, hydrogen bonding and inclusion complexation with chiral analytes, and therefore resulting in efficient enantioseparations. Systematic experiments were performed to investigate the effects of concentration of CS, composition of background electrolyte (BGE) and applied voltage on chiral separation. Enantiomers of propranolol and metoprolol were baseline resolved using a BGE consisting of 20mM CS and 50/50 (v/v) iso-propanol/phosphate buffer (100mM, pH 7.0) whereas for enantiomers of sertraline, a BGE consisting of 23mM CS and 40/60 (v/v) iso-propanol/phosphate buffer (100mM, pH 7.0) resulted in baseline resolutions.

Enantioseparation of basic chiral drugs on a carbamoylated erythromycin-zirconia hybrid monolith using capillary electrochromatography.

An organic-inorganic hybrid monolithic column was prepared within the confines of a capillary via a single-step in situ sol-gel approach using zirconium tetrabutoxide as a precursor to compose the inorganic backbone and 3-triethoxysilylpropyl carbamoylated derivative of erythromycin (TEOSPC-ERY) as a co-precursor to introduce the organic chiral selector moiety in the zirconia backbone. The resulting carbamoylated ERY-zirconia hybrid monolith (ERY-ZHM) showed homogeneous morphology with well-defined through pores and was tightly connected with the inner wall of the capillary. The column was employed for capillary electrochromatographic enantioseparation of six basic chiral drugs in mobile phases (MPs) consisting of acetonitrile (ACN) and triethylammonium acetate (TEAA) buffer. The effects of composition of MP and applied voltage on chiral separation were investigated by using propranolol as a representative analyte. The highest resolution (Rs=3.33) was obtained with a MP consisting of 10/90 (v/v) ACN/TEAA buffer (10mM, pH 7), 10 kV applied voltage and 25°C capillary temperature. The relative standard deviations for resolution values regarding run to run, day to day, column to column and batch to batch repeatability were 0.41%, 0.89%, 1.80% and 2.26% (for n=3), respectively, indicating satisfactory stability of columns and reproducibility of column preparation process.

Enantiomer separation of acidic chiral compounds on a quinine-silica/zirconia hybrid monolith by capillary electrochromatography.

A weak anion-exchanger chiral selector, quinine-incorporated silica/zirconia hybrid monolithic (QUI-S/ZHM) capillary column was prepared by sol-gel technology. The performance of the QUI-S/ZHM column was investigated for enantioresolution of a set of acidic chiral drugs and dinitrobenzoyl (DNB)-amino acids by capillary electrochromatography in aqueous organic mobile phases composed of acetonitrile (ACN) and triethylammonium acetate (TEAA) buffer. Effects of several parameters including the ACN content, concentration and pH of the mobile phase on the chiral separation were examined. Baseline resolutions of all the compounds were obtained in the mobile phase consisting of 70:30 ACN/TEAA (10mM, pH 6) under applied voltage of -10kV at 25°C within 20min.

Enantioseparation of basic chiral compounds on a clindamycin phosphate-silica/zirconia hybrid monolith by capillary electrochromatography.

An organic-inorganic silica/zirconia hybrid monolithic capillary column was prepared by sol-gel process in a fused-silica capillary by using triethoxysilylpropylcarbamate (TEOSPC) derivative of clindamycin phosphate (CLIP) as a chiral selector. A sol solution consisting of 6 × 10(-3)M of polyethylene glycol, 1 M of water, 2M of acetic acid and 0.04/0.96 ratio of CLIP-TEOSPC/Zr-Bu resulted in homogeneous monolith having well defined through-pores and tightly anchored to the capillary wall. The column was employed for capillary electrochromatographic enantioseparation of eight basic chiral drugs in mobile phases consisting of acetonitrile, methanol and ammonium acetate (AA, as the electrolyte). Effects of the compositions of solvents and electrolyte in the mobile phase, applied voltage and capillary temperature on chiral separation were investigated. The highest resolution values were obtained with mobile phases consisting of 40/60 MeOH/ACN and 100 mM AA (for citalopram, Tröger's base, indapamide, metoprolol, cetirizine and atropine) and 35/65 MeOH/ACN and 100 mM AA (for sertraline and propranolol) using -10 kV applied voltage at 25 °C.

Enantiomer separations of basic chiral compounds by capillary electrochromatography on a phosphated ß-cyclodextrin-modified zirconia monolith.

Phosphated ß-cyclodextrin (PCD)-coated zirconia monolith was used as the chiral stationary phase in capillary electrochromatography (CEC) for separation of four basic chiral compounds including metoprolol (MET), sertraline (SER), citalopram (CIT) and atenolol (ATE). Migration, chiral selectivity and resolution data were measured in reversed-phase mobile phases of varying pH, buffer and organic modifier compositions. Optimum mobile phase conditions for CEC separation of the compounds studied were found to be a 15-mM aqueous buffer of pH 5.0 with 5mM PCD. Baseline separations of enantiomers of CIT, MET and SER, and partial separation for ATE were achieved with the optimal mobile phase.

Application of antibiotics as chiral selectors for capillary electrophoretic enantioseparation of pharmaceuticals: a review.

Recent years have witnessed several new trends in chiral separation, for example, the enantiorecognition ability of several new antibiotics has been explored using capillary electrophoresis (CE) prior to HPLC; antibiotics have been employed as chiral selectors (CSs) in a nonaqueous CE (NACE) mode; and several new detection techniques (namely, capacitively coupled contactless conductivity detection) have been used in combination with CE for quantification of enantiomers. On account of these emerging trends, this article aims to review the application of various classes of antibiotics for CE enantioseparation of pharmaceuticals. A detailed account of the basic factors affecting enantioseparation, certain limitations of antibiotics as CSs and strategies to mitigate them, and advantages of NACE while using antibiotics as CSs has also been presented.

Penicillin G as a novel chiral selector in capillary electrophoresis.

The penicillin sub-class of ß-lactam antibiotics has not been examined for its enantiodiscriminating abilities in capillary electrophoresis (CE) until date. The present work was therefore designed to evaluate penicillin G potassium salt (PenG) as an ion-pair chiral selector (CS) using CE for its several attributes, namely, high solubility in water and lower alcohols, structure allowing multiple interactions with analytes and cost-effectiveness. Systematic experiments were performed to investigate the effect of composition of background electrolyte, applied voltage and capillary temperature on chiral separation. Baseline resolutions of enantiomers of five basic chiral drugs (namely, darifenacin, citalopram, sertraline, propranolol and metoprolol) were attained using a background electrolyte composed of water:methanol (90:10, v/v) and consisting of 10.7 or 16.1mM CS at 20°C using an applied voltage of 5kV.

Enantioseparation of chiral acids and bases on a clindamycin phosphate-modified zirconia monolith by capillary electrochromatography.

Porous zirconia monolith modified with clindamycin phosphate (CLIP-ZM) was used as chiral stationary phase (CSP) to separate a set of six acidic and basic chiral compounds in capillary electrochromatography (CEC). Resolutions and chiral selectivity factors of the chiral compounds were measured in ACN/MeOH mobile phases of varying compositions of MeOH and ammonium acetate as the electrolyte. In contrast to the CE separations where only chiral separations of acidic compounds were achieved enantiomers of both acidic and basic compounds were separated with acidic compounds better resolved than basic ones by CEC on the CLIP-ZM CSP. Best chiral separations of the chiral compounds were obtained with an eluent containing 10mM ammonium acetate and 35 vol.% methanol.

Chiral separation of basic compounds on a cellulose 3,5-dimethylphenylcarbamate-coated zirconia monolithin basic eluents by capillary electrochromatography.

Porous zirconia monolith (ZM) modified with cellulose 3,5-dimethylphenylcarbamate (CDMPC) was used as chiral stationary phase to separate basic chiral compounds in capillary electrochromatography. The electroosmotic flow behavior of bare and CDMPC-modified zirconia monolithic (CDMPC-ZM) column was studied in ACN/phosphate buffer eluents of pH ranging from 2 to 12. The CDMPC-ZM column was evaluated by investigating the influences of pH, the type and composition of organic modifier of the eluent on enantioseparation. CEC separations at pH 9 provided the best resolutions for the analytes studied, which are better than those observed on CDMPC-modified silica monolithic columns under similar chromatographic conditions. No appreciable decline in retention and resolution factors after over 200 injections, and run-to-run and day-to-day repeatabilities of the column of less than 3% indicate the stability of the zirconia monolithic column in basic media.

Fast separations of chiral ß-blockers on a cellulose tris(3,5-dimethylphenylcarbamate)-coated zirconia monolithic column by capillary electrochromatography.

Cellulose tris(3,5-dimethylphenylcarbamate) (CDMPC) is an excellent chiral selector for enantioseparation of a wide variety of chiral compounds. The monolithic chiral columns are becoming popular in liquid chromatography and capillary electrochromatography. In this work, we present the fast separation of chiral ß-blockers on a CDMPC-modified zirconia monolithic column by capillary electrochromatography (CEC). The porous zirconia monolithic capillary column was prepared by using the sol-gel technology and then zirconia surface modified with CDMPC. The enantioseparations were performed in reversed-phase (RP) eluents of a phosphate solution (pH 4.4) modified with acetonitrile or alcohol. The enantioseparations of a set of eight chiral ß-blockers were achieved in less than one minute. Influences of the applied voltage, column temperature, concentration of acetonitrile and the type of alcohol as the organic modifier in the mobile phase, and sample injection time on enantioseparation were investigated. CEC separations at the applied voltage of 10 kV and 15 °C in the ACN-modified mobile phase provided the best resolutions for the analytes studied. Run-to-run and day-to-day repeatabilities of the column in the RP-CEC separation were less than 1 and 2%, respectively.

Azithromycin as a new chiral selector in capillary electrophoresis.

In capillary electrophoresis (CE), separation of enantiomers of a chiral compound can be achieved through the chiral interactions and/or complex formation between the chiral selector and the enantiomeric analytes on leaving their diastereomeric forms with different stability constants and hence different mobilities. A great number of chiral selectors have been employed in CE and among them macrocyclic antibiotics exhibited excellent enantioselective properties towards a wide number of racemic compounds. The use of azithromycin (AZM) as a chiral selector has not been reported previously. This work reports the use of AZM as a chiral selector for the enantiomeric separations of five chiral drugs and one amino acid (tryptophan) in CE. The enantioseparation is carried out using polar organic mixtures of acetonitrile (ACN), methanol (MeOH), acetic acid and triethylamine as run buffer. The influences of the chiral selector concentration, ACN/MeOH ratio, applied voltage and capillary temperature on enantioseparation are investigated. The results show that AZM is a viable chiral selector in CE for the enantioseparation of the type of chiral drugs investigated.

Enantioseparation on cellulose dimethylphenylcarbamate-modified zirconia monolithic columns by reversed-phase capillary electrochromatography.

This work reports the preparation of monolithic zirconia chiral columns for separation of enantiomeric compounds by capillary electrochromatography (CEC). Using sol-gel technology, a porous monolith having interconnected globular-like structure with through-pores is synthesized in the capillary column as a first step in the synthesis of monolithic zirconia chiral capillary columns. In the second step, the surface of the monolith is modified by coating with cellulose tris(3,5-dimethylphenylcarbamate) (CDMPC) as the chiral stationary phase to obtain a chiral column (CDMPCZM). The process of the preparation of the zirconia monolithic capillary column was investigated by varying the concentrations of the components of the sol solution including polyethylene glycol, water and acetic acid. CDMPCZM is mechanically stable and no bubble formation was detected with the applied current of up to 30 microA. The enantioseparation behavior of the CDMPCZM columns was investigated by separating a set of 10 representative chiral compounds by varying the applied voltage and pH and organic composition of the aqueous organic mobile phases.

Enantioseparation of alpha-amino acids on an 18-crown-6-tetracarboxylic acid-bonded silica by capillary electrochromatography.

(-)-(18-Crown-6)-2,3,11,12-tetracarboxylic acid-bonded silica was used as the chiral stationary phase in capillary electrochromatography (CEC) for enantioseparation of some alpha-amino acids. Separation data in CEC were measured in mobile phases of varying pH, and composition of methanol and buffer, and compared with those in capillary liquid chromatography (CLC). In CEC better enantioseparation was generally obtained in the eluent of lower pH, higher buffer concentration and intermediate MeOH content, usually at the expense of analysis time. CEC showed generally better enantioselectivity and resolutions than CLC for the amino acids investigated.

Cellulose dimethylphenylcarbamate-immobilized zirconia for chiral separation in reversed-phase CEC.

Cellulose dimethylphenylcarbamate (CDMPC)-immobilized zirconia (CDMPCZ) was used as a chiral stationary phase for enantioseparation of a set of nine racemic compounds in reversed-phase CEC. Influences of the type and composition of organic modifier and the applied voltage on enantioseparation were examined. Separation data on CDMPCZ were also compared with those on CDMPC-immobilized silica (CDMPCS). Enantiomers of the analytes investigated are well separated in ACN/phosphate buffer mobile phases. Better enantioselectivity and resolution were obtained with ACN than MeOH as the organic modifier. Retention was longer but better enantioselectivity and resolution were obtained on CDMPCZ than CDMPCS.

Chiral separation of amides of amino acid on a (S)-N-(3,5-dinitrobenzoyl)leucine-N-phenyl-N-propylamide-bonded silica using nonaqueous capillary electrochromatography.

(S)-N-(3,5-dinitrobenzoyl)leucine-N-phenyl-N-propylamine-bonded silica was used as a chiral stationary phase for separation of a set of racemic pi-acidic and pi-basic alpha-amino acid amides in electrolyteless ACN-water eluents by CEC in the RP and polar organic (PO) modes. The effect of the amount of water in the ACN-water eluent on chiral separation was examined. As water is added to ACN, retention was shortened but resolution and selectivity deteriorated severely. Retention, enantioselectivity, and resolution factors obtained in 100% ACN were compared with those in an n-hexane-isopropanol eluent with a small amount of water by normal phase (NP) CEC. Much shorter retention times with comparable enantioselectivities were observed with 100% ACN, demonstrating the advantage of separation on (S)-N-(DNB)leucine-N-phenyl-N-propylamine-bonded silica in PO-CEC over NP-CEC.

Enantioseparation of amino acid amides on an (S)-N-(3,5-dinitrobenzoyl)leucine-N-phenyl-N-propylamide-bonded silica in normal phase CEC.

(S)-N-(3,5-Dinitrobenzoyl)leucine-N-phenyl-N-propylamide-bonded silica was used as a chiral stationary phase for separation of enantiomers of some racemic pi-acidic and pi-basic amino acid amides in normal phase capillary LC (CLC) and CEC. For generation of EOF in CEC, different amounts of water were added to n-hexane-isopropanol (IPA) eluents. Influences of added water and composition of polar modifier on retention, enantioselectivity and resolution were studied. Generally better separation was obtained in the eluents of lower water content. Shorter retention and better enantioselectivities were observed in the eluents of higher IPA content, whereas resolution became lower as IPA content was increased. Enantioselectivities were comparable but resolutions were much better in CEC than CLC.

Cellulose dimethylphenylcarbamate-bonded carbon-clad zirconia for chiral separation in high performance liquid chromatography.

Porous zirconia particles are very robust material and have received considerable attention as a stationary phase support for HPLC. We prepared cellulose dimethylphenylcarbamate-bonded carbon-clad zirconia (CDMPCCZ) as a chiral stationary phase (CSP) for separation of enantiomers of a set of 14 racemic compounds in normal phase (NP) and reversed-phase (RP) liquid chromatography. Retention and enantioselectivity on CDMPCCZ were compared to those on CDMPC-coated zirconia (CDMPCZ) to see how the change in immobilization method of the chiral selector affects the retention and chiral selectivity. In NPLC, retention was longer and the number of resolved racemates was smaller on CDMPCCZ than on CDMPCZ. However, chiral selectivity factors for some resolved racemates were better on CDMPCCZ than on CDMPCZ. The longer retention on CDMPCCZ is likely due to strong, non-chiral discriminating interactions with the carbon layer on CDMPCZ. In RPLC only two racemates were resolved on CDMPCCZ, but retention times were shorter than, and resolutions were comparable to, those in NPLC, indicating a potential for improving chromatographic performance of the CDMPCCZ column in RPLC with optimized column preparation and separation conditions.

Intermolecular interactions on multiwalled carbon nanotubes in reversed-phase liquid chromatography.

Retention on multiwalled carbon nanotubes (MWCNTs) in RPLC has been correlated with solute descriptors of dispersion, polarizability, dipolarity, hydrogen bond donor acidity, and hydrogen bond acceptor basicity through the use of the linear solvation energy relationship. Intermolecular interactions influencing solute retention on MWCNTs were compared with those on a graphitic carbon-deposited zirconia and a common RPLC stationary phase, octylsilane-bonded silica.