Synthesis of novel chiral stationary phases for high-performance liquid chromatography

Three novel chiral selectors 4a-c were synthesized from(S)-amino acids and(R)-1-phenyl-2-(4-methylphenyl)ethylamine.4a-cwere connected to 3-aminopropylsilanized silica gel to be used as the chiral stationary phase for HPLC.Five amino acid derivativesand two pyrethroid insecticides were fairly resolved on these three new chiral stationary phases under normal phase condition.     

Diastereo-enantioseparation of novel γ-lactamic derivatives on cellulose chiral stationary phases

Summary This paper describes the enantiorecognition of 1-methyl-3-hydroxy-5-aryl-2-pyrrolidinonic systems by high-performance liquid chromatography using two chiral derivatized cellulose stationary phases (CSPs) operated in the normal phase mode. According our results, the tris-(3,5-dimethylphenyl carbamate) cellulose, Chiralcel^? OD, is more suitable than the tris-(4-methyl-phenylbenzoate), Chiralcel^? OJ. On the first column, the resolution of the pyrrolidinonic compounds depends on the alcoholic modifier percentage. A possible solute-stationary phase recognition mechanism is discussed. The temperature and mobile phase composition have been considered to explain the different contribution for the enantiomeric resolution.     

Stellan Hjertén's contribution to the development of monolithic stationary phases

This overview is presented to celebrate the birthday of one of the luminaries of the separation science and my friend - Stellan Hjertén. He made significant contributions to a variety of areas in separation science such as electrophoresis, LC, and CEC to name just a few. Since the scope of his work was enormous, this review will focus only on a single aspect of his scientific activities, the design and applications of monolithic materials. During the years starting from 1989, Stellan Hjertén published many excellent papers concerning the preparation of acrylamide chemistry-based monoliths and their use in both micro-HPLC and CEC. The following text details his works in the field.     

Chiral metal–organic framework used as stationary phases for capillary electrochromatography

Metal–organic frameworks (MOFs) have received great attention as novel media in separation sciences because of their fascinating structures and unusual properties. However, to the best of our knowledge, there has been no attempt to utilize chiral MOFs as stationary phases in capillary electrochromatography (CEC). In this study, a homochiral helical MOF [Zn₂(D-Cam)₂(4,4′-bpy)]_n (D-Cam = D-(+)-camphoric acid, 4,4′-bpy = 4,4′-bipyridine) was explored as the chiral stationary phase in open tubular capillary electrochromatography (OT-CEC) for separation of chiral compounds and isomers. The MOFs coated column has been developed using a simple procedure via MOFs post-coated on the sodium silicate layer. The baseline separations of flavanone and praziquantel were achieved on the MOFs coated column with high resolution of more than 2.10. The influences of pH, organic modifier content and buffer concentration on separation were investigated. Besides, the separations of isomers (nitrophenols and ionones) were evaluated. The relative standard deviations (RSDs) for the retention time of run-to-run, day-to-day and column-to-column were 1.04%, 2.16% and 3.07%, respectively. The results demonstrated that chiral MOFs are promising for enantioseparation in CEC.     

Retention properties of novel β-CD bonded stationary phases in reversed-phase HPLC mode

With the given special structures, the CD bonded stationary phases are expected to have complementary retention properties with conventional C18 stationary phase, which will be helpful to enhance the polar selectivity in RP mode separation. In this work, two β-cyclodextrin (β-CD) bonded stationary phases for reversed-phase HPLC, including 1, 12-dodecyldiol linked β-CD stationary phase (CD1) and olio (ethylene glycol) (OEG) linked β-CD stationary phase (CD2), have been synthesized via click chemistry. The resulting materials were characterized with FT-IR and elemental analysis, which proved the successful immobilization of ligands. The similarities and differences in retention characteristics between the CD and C18 stationary phases have been elucidated by using comparative linear solvation energy relationships (LSERs). The force related to solute McGowan volume has no significant difference, while the hydrogen bonding and dipolar interactions between solutes and CD stationary phases are stronger than between solutes and C18, which is attributed to the special structures (CD and triazole groups) of CD stationary phases. Chemical origins are interpreted by comparison between CD1 and CD2. Similar dispersive interactions of CD1 and CD2 are attributed to their similar length of spacer arms. CD2 which contains OEG spacer arm has relative weaker HBD acidity but stronger HBA basicity. CD stationary phases display no serious different methylene selectivity and higher polar selectivity than in the case of C18. Higher acid selectivity and lower basic selectivity are observed on CD2 than on CD1. Distinctive retention properties and good complementary separation selectivity to C18 make the novel CD bonded stationary phases available for more application in RPLC.     

Preparation and evaluation of tandem stationary phases for μ-HPLC and capillary electrochromatography

Tandem separation beddings were prepared within the capillary tube with a photopolymerized monolith initially formed by sol-gel technology in combination with microspheric octadecylsilane material by slurry packing. The chromatographic performance of the tandem stationary phases was evaluated in detail by varying the flow rate and the composition of the mobile phase using a self-installed capillary HPLC system. For the tandem stationary phases with a forepart monolithic length of 11 cm and 1 cm the lowest theoretical plate height for the retained component was 26 µm and 34 µm, respectively. After evaluation by capillary electrochromatography, enhanced chromatographic performance was obtained using a column with 1 cm monolithic inlet frit with a theoretical plate height up to 7.20 µm. A scanning electron micrograph with different cross-sections of the column showed that a porous network formed in the center of the capillary and a homogenous slurry packing of C₁₈ was obtained at the back part.     

Characterization and classification of stationary phases in HPLC and SFC – a review

Packed column supercritical fluid chromatography (pSFC) is an attractive technique in drug discovery related analysis because it offers several advantages over the more commonly used high-performance liquid chromatography (HPLC) technique. The environmental-friendly CO₂ mobile phase, the high-throughput capacity, the increased efficiency and the lower operational costs give SFC additional benefits over HPLC in analysis related to drug development. The latter technique is well established and has been used for decades in the pharmaceutical industry. On the other hand, SFC is still in its infancy, even though the technique has been known for decades and researchers are still discovering the possibilities and limitations of this technique.     

In-column “click” preparation of hydrophobic organic monolithic stationary phases for protein separation

Two types of macroporous organic polymer monoliths based on glycidyl methacrylate (GMA), 4-vinylbenzyl chloride (VBC) and divinylbenzene (DVB) were prepared inside stainless-steel tubes. Azide functionalities were firstly introduced on the surfaces of poly(GMA-co-DVB) and poly(VBC-co-DVB) monoliths to provide reactive sites for click chemistry. With the application of copper(I)-catalyzed (3 + 2) azide-alkyne cycloaddition, an in-column click-modification approach for covalent attachment of long alkyl chains onto polymer monoliths was developed. The column morphology and surface chemistry of the fabricated monolithic columns were characterized by the scanning electron microscopy, mercury intrusion porosimeter, Fourier transform infrared spectroscopy, and elemental analyses, respectively. The chromatographic performances of the “clicked” stationary phases were demonstrated with the high separation efficiency for a variety of proteins within 4 min.     

Kováts' coefficients for predicting polarities in silicone stationary phases and their mixtures

Summary A correlation between the retention polarity and the Kováts' coefficient has been found for a number of commercial stationary phases used in gas chromatography. The correlation has been used to predict the polarity of a few mixed stationary phase columns prepared as binary mixtures of OV-101 with OV-25, DC-550 or Carbowax 20M, and also mixtures of OV-225 with SP-2340. A linear relationship was found between polarity and mixture composition. The temperature dependence of the Kováts' coefficient was investigated and found to increase linearly with temperature, the rate of increase depending on the polarity of the stationary phase, with greater increases for more polar stationary phases.     

β-Cyclodextrin covalent organic framework supported by polydopamine as stationary phases for electrochromatographic enantioseparation

In this work, a new open-tubular capillary electrochromatography (OT-CEC) column was prepared using β-cyclodextrin covalent organic framework (β-CD COF) as a stationary phase. Polydopamine was used to assist fabrication of β-CD COF on an inner wall of a fused-silica capillary. The coating layer on the capillary was characterized by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). Electroosmotic flow (EOF) was also studied to evaluate the variation of the inner wall of immobilized columns. Furthermore, the chiral separation effectiveness of the fabricated capillary column was evaluated by CEC using enantiomers of several related proton pump inhibitors as model analytes, including omeprazole, lansoprazole, pantoprazole and tenatoprazole. The effects of bonding time and concentration of β-CD COF, the type, concentration and pH of buffer, applied voltage were investigated to obtain satisfactory enantioselectivity. In the optimum conditions, the enantiomers of four analytes were resolved within 15 min with resolutions of 1.63–2.62. The relative standard deviation values for migration times and resolutions of the analytes representing intraday and interday were less than 6.75% and 4.24%, respectively. The results reveal that β-CD COF has great potential as chiral-stationary phases for enantioseparation in CEC.     

Preparation of new hybrid organic/inorganic polymeric chiral stationary phases for ligand-exchange chromatography

Three new hybrid organic/inorganic polymeric ligand-exchange chiral stationary phases were developed by radical chain transfer reaction and surface grafting on silica gel,and successfully used for the enantioseparations of DL-amino acids and DL-hydroxyl acids.The resolutions were achieved by using water containing 2.0×10~(-4) mol/L of CuAc_2 as a mobile phase,column temperature of 40℃,flow rate of 1.0mL/min and detection at UV 254 nm.The elution order of D-isomer before L-isomer was observed for all DL-amino acids resolved except DL-Pro.     

Synthesis and mechanistic study of novel π-basic chiral stationary phases derived from tyrosine

Summary This work was a study of novel -basic, chiral stationary phases (CSPs) deriving from tyrosine and bearing two stereogenic centres. These CSPs differ in the nature of the amidic substituent (n-butyl ortert-butyl) and by the configuration of both the chiral centres (S or R). The enantioselectivity and elution order of various 3, 5-dinitrobenzoyl derivatives were studied using liquid and supercritical fluid chromatographic modes. The chromatographic data allowed determination of the influence of the CSP structure (also studied according to factorial designs) and determination of the influence of the solute structure. Chiral recognition mechanisms which are in good agreement with these observations are proposed. Finally, this paper reports the enantiomeric resolution of two compounds of pharmaceutical interest (warfarin and ICI 176334).     

Metal organic framework–organic polymer monolith stationary phases for capillary electrochromatography and nano-liquid chromatography

In this study, metal organic framework (MOF)–organic polymer monoliths prepared via a 5-min microwave-assisted polymerization of ethylene dimethacrylate (EDMA), butyl methacrylate (BMA), and 2-acrylamido-2-methylpropane sulfonic acid (AMPS) with the addition of various weight percentages (30–60%) of porous MOF (MIL-101(Cr)) were developed as stationary phases for capillary electrochromatography (CEC) and nano-liquid chromatography (nano-LC). Powder X-ray diffraction (PXRD) patterns and nitrogen adsorption/desorption isotherms of these MOF–organic polymer monoliths showed the presence of the inherent characteristic peaks and the nano-sized pores of MIL-101(Cr), which confirmed an unaltered crystalline MIL-101(Cr) skeleton after synthesis; while energy dispersive spectrometer (EDS) and micro-FT-IR spectra suggested homogenous distribution of MIL-101(Cr) in the MIL-101(Cr)–poly(BMA–EDMA) monoliths. This hybrid MOF–polymer column demonstrated high permeability, with almost 800-fold increase compared to MOF packed column, and efficient separation of various analytes (xylene, chlorotoluene, cymene, aromatic acids, polycyclic aromatic hydrocarbons and trypsin digested BSA peptides) either in CEC or nano-LC. This work demonstrated high potentials for MOF–organic polymer monolith as stationary phase in miniaturized chromatography for the first time.     

Separation of racemiccloso-3,3-(η3,2-norbornadienyl)rhodacarboranes into enantiomers by HPLC on chiral stationary phasesinto enantiomers by HPLC on chiral stationary phases

Racemiccloso-rhodacarboranes,vis. closo-(η^3,2-C₇H₃-2-CR ₂ ¹ )-1-R²-2-R³-3,1,2-RhC₂B₉H₉ (R¹=R²=R³=H; R¹=H, R²=R³=Me; R¹=R²=R³=Me) and (closo-2,2-(η^3,2-C₇H₇-2-CH₂)-2,1,7-RhC₂B₉H₁₁), were successfully separated into enantiomers by high-performance liquid chromatography (HPLC). Translated fromIzvestiya Akademii Nauk. Seriya Khimicheskaya, No. 4, pp. 759–761, April, 2000.     

Preparation and enantioseparation characteristics of three chiral stationary phases based on modified β-cyclodextrin for liquid chromatography

In order to study the effect of the nature and the length of the spacer, three mixed 10-undecenoate/phenylcarbamate derivatives of β-cyclodextrin have been prepared and linked to allylsilica gel by means of a radical reaction. The chiral recognition ability of the resulting materials, when used as liquid chromatography chiral stationary phases (CSPs), was evaluated using heptane and either 2-propanol or chloroform as organic mobile-phase modifiers. A large variety of racemic compounds have been separated successfully on these CSPs (mainly pharmaceuticals and herbicides). Optimization of these separations was discussed in terms of mobile-phase composition and structural patterns of the injected analytes. The efficiencies of the three prepared materials were compared to those of previously described perphenylated-β-cyclodextrin column and to analogous cellulose derivative-based CSPs. Schematic illustration of the b-cyclodextrin/mandelic acid inclusion complex     

Novel β-cyclodextrin chiral stationary phases with different length spacers for normal-phase high performance liquid chromatography enantioseparation

Cyclodextrin and its derivatives are widely used as selectors of chiral stationary phases (CSPs) for high performance liquid chromatography (HPLC) due to their unique molecular structure and resolution capability. Three mono(6(A)-N-(ω-alkenylamino)-6(A)-deoxy)perphenylcarbamoylated β-cyclodextrin (PICD) based CSPs with different length spacers have been prepared, with their enantioseparation abilities evaluated with 10 model racemates including aromatic alcohols, flavanone compounds, amine and non-protolytic compounds under normal-phase conditions. The effect of spacer length and surface loading on the enantioseparation performance of CSPs is investigated herewith. The results indicate that higher surface loading 6C-PICD displays the best enantioselectivities towards selected racemates under normal-phase conditions.