Streptavidin chiral stationary phase for the separation of adenosine enantiomers

In this paper, a microbore column packed with streptavidin particles was used, at various temperatures (0-24 degrees C), to separate the adenosine enantiomers by HPLC. Using an aqueous mobile phase, the apparent enantioseparation was high for a small molecule, varying from 11.5 at 0 degrees C to 6.2 at 24 degrees C. From the experiments carried out with a streptavidin-biotin complex stationary phase, it was demonstrated that the blockage of the biotin sites of the immobilized streptavidin was responsible for a strong decrease in the enantioselectivity via a direct and/or an indirect effect. From the analysis of the concentration dependencies of the solute retention factor, it was also shown that a reduction of the D-adenosine specific binding sites occurred at the lowest temperature. The thermodynamic parameters determined from the van't Hoff plots indicated that the D-adenosine binding to the streptavidin specific sites was enthalpically driven.     

Separation of the stereoisomers of racemic fluoroquinolone antibacterial agents on a crown-ether-based chiral HPLC stationary phase

Summary A chiral stationary phase derived from (+)-(18-crown-6)-2,3,11,12-tetracarboxylic acid has been successfully used for the direct separation of the enantiomers of recemic fluoroquinolines containing a primary amino group being investigated as antibacterial agents. The chromatographic resolution behavior was found to depend on the content and the type of acidic and organic modifiers in the mobile phase and on the column temperature.     

Preparation of a new crown ether-based chiral stationary phase containing thioester linkage for the liquid chromatographic separation of enantiomers

A new chiral stationary phase (CSP) containing thioester linkages was prepared by bonding (+)-(18-crown-6)-2,3,11,12-tetracarboxylic acid to mercaptopropylsilica gel. The chiral recognition ability of the new CSP was found to be greater than that of the previously reported CSP containing amide linkages in the resolution of the various α-amino acids that were tested, except for that of Met, Ser and Thr. In the resolution of racemic amines and amino alcohols, the new CSP was always better than the one containing amide linkages in terms of the separation factors (α) and the resolutions (R_S). Given the identical elution orders on the two CSPs, it was concluded that the chiral recognition mechanism is not affected by the change of the linkage type. In addition, the new CSP was found to be quite stable under the acidic mobile phase conditions that were utilized, indicating that the thioester linkage is useful as a tethering group.     

Separation of the enantiomers of pyrethroic acids and their esters by high-performance liquid chromatography on a polysaccharide-derived chiral stationary phase

Summary The liquid-chromatographic separation of the enantiomers of pyrethroic acids and their esters has been investigated on a polysaccharide-derived chiral stationary phase (CSP), Chiralpak AS. Good separation of the enantiomers of underivatized pyrethroic acids was achieved on the column, and the enantiomers of pyrethroic acid methyl and ethyl ester derivatives were also resolved.     

The preparative chromatographic enantioseparation of a racemic morphanthridine analog on a chiral stationary phase

Summary The preparative chromatographic enantioseparation of a chiral morphoanthridine analog has been performed on an analytical column using amylose-tris(3,5-dimethylphenylcarbamate) as chiral stationary phase. The racemate (100 mg) was resolved to baseline within 15 min. This paper describes the development of the method, estimation of the capacity of the chiral stationary phase and discussed the potential of the chromatography if performed under preparative conditions. From the results and calculations presented it seems likely that the resolution of 70 tons year⁻¹ could easily be achieved on 30 kg of stationary phase with a mobile-phase consumption of only 720 L day⁻¹.     

Polymethacrylate-type monoliths functionalized with chiral amino phosphonic acid-derived strong cation exchange moieties for enantioselective nonaqueous capillary electrochromatography and investigation of the chemical composition of the monolithic polymer

In situ prepared monolithic poly(glycidyl methacrylate-co-ethylene dimethacrylate) (poly(GMA-co-EDMA)) capillary columns were activated to reactive thiol-monoliths and subsequently functionalized with (S)-N-(4-allyloxy-3,5-dichlorobenzoyl)-2-amino-3,3-dimethylbutanephosphonic acid as chiral selector by radical addition to afford enantioselective strong cation exchanger (SCX) capillary columns (100 microm inner diameter (ID)). These monolithic capillaries were devised for the enantioseparation of chiral bases by nonaqueous and aqueous capillary electrochromatography (CEC) and the results obtained for mefloquine and its tert-butylcarbamate as test compounds were compared to those obtained with particulate silica-based analogs (packed columns). Despite abolishment of nonspecific ionic interactions between the cationic solutes and residual silanols that may diminish separation factors of the silica-based chiral SCX particles, the poly(GMA-co-EDMA)-supported SCX monolith did not, as expected, show better enantioselectivities, which was assumed to be due to detrimental nonspecific interactions between the analytes and the lipophilic polymer backbone. In order to minimize these unfavorable contributions, less lipophilic monoliths were developed by copolymerization of different amounts of the hydrophilic monomer 2-hydroxyethyl methacrylate (HEMA) with GMA and EDMA, leading to GMA-co-HEMA-co-EDMA-terpolymeric monoliths. By this increase of the hydrophilicity of the monolithic support the enantioselectivity of the resultant SCX stationary phase could be enhanced and reached values comparable to the packed silica-based enantioselective SCX capillaries. Additionally, the mobile phase composition and other variables were examined and it could be shown that the separation factors are considerably affected by diverse parameters such as acetonitrile-methanol ratio and type and concentration of the counterion. Mefloquine enantiomers could be separated with alpha-values up to 1.56 and a maximum plate count of ca. 60,000 m(-1) could be achieved.     

Comparison of different heptakis(6-O-alkyldimethylsilyl-2-3-di-O-ethyl)-β-cyclodextrins as chiral stationary phases in capillary GC

Three new β-cyclodextrin derivatives, heptakis(6-O-isopropyldi-methylsilyl-2,3-di-O-ethyl)-β-cyclodextrin, heptakis(6-O-thexyldi-methylsilyl-2,3-di-O-ethyl)-β-cyclodextrin, and heptakis(6-O-cy-clohexyldimethyl-2,3-di-O-ethyl)-β-cyclodextrin (IPDE-β-CD, TXDE-β-CD, and CHDE-β-CD), were synthesized and the enan-tioselectivities of these three CD derivatives and heptakis(6-O-tert-butyldimethylsilyl-2,3-di-O-ethyl)-β-cyclodextrin (TBDE-β-CD) were compared for GC separation of a range of chiral test com-pounds. In particular TXDE-β-CD showed much higher enentio-selectivity than TBDE-β-CD. Enentioselectivities of IPDE-β-CD and CHDE-β-CD are somewhat lower than that of TXDE-β-CD and CHDE-β-Cd are somewhat lower than that of TXDE-β-CD. These observations are indicative of significant effects of subtle changes in the structure of the 6-O-substituent on the enantioselec-tivity of the β-CD derivatives. The difference in enantioselectivities of the 6-O-substituted CD derivatives were explained in terms of relative contributions of the effects of hydrophobicity and steric hindrance of the substituent to the inclusion process. CHDE-β-CD showed the lowest enantioselectivity among the threederivatives. It is likely that the unfavorable steric hindrance of the bulky cyclo-hexyl group plays a greater role than the favorable hydrophobicity effect of the cyclohexyl group in the inclusion process in CHDE-β-CD. IPDE-β-CD showed lower selectivity than TXDE-β-CD and TBDE-β-CD. In the case of these CD derivatives having acyclic substituents the relative hydrophobicity of the substituent seems to be a dominant factor affecting the inclusion process. Isopropyl groups factor affecting the inclusion process. Isopropyl groups are less hydrophobic than thexyl and tert-butyl groups.     

Influence of the structure of organic phosphonate compounds on chiral separation on a pirkle type chiral stationary phase

Summary The enantiomers of eightO,O-dialkyl-2-benzyloxycarbonyl-aminoarylmethyl-phosphonates are directly separated on anN-(3,5-dinitrobenzoyl)leucine (DNBleu) column. The influence of the mobile phase composition and the column temperature on the retention and the enantioselectivity are investigated. The influence of the length and steric hindrance of alkoxyl groups of the phosphonate ester and the nature of the substituentp-Cl and pH on the benzene ring which is attached to the chiral carbon atom on chiral separation are discussed also.     

Effect of the structure of organic phosphonate compounds on chiral separations on derivatized cellulose chiral stationary phase

Summary The enantiomers of eightO,O-dialkyl-2-benzyloxycarbonylaminoarylmethyl phosphonates have been directly separated on a tris(3,5-dimethylphenylcarbamate) cellulose column. The results are very different from those obtained by separation on anN-(3,5-dinitrobenzoyl)leucine (DNBleu) column. The effect of mobile phase composition and column temperature on retention and enantioselectivity were investigated. The effect on chiral separation of the length of, and steric hindrance by, alkoxyl groups of the phosphonate ester and of the nature of the substitutentsp-Cl andp-H on the benzene ring attached to the chiral carbon atom are also discussed.     

An improved version of an old fluorocarbinol chiral stationary phase

A chiral stationary phase (CSP) derived from anthrone is quite effective for the chromatographic resolution of the enantiomers of a wide variety of compounds. Denoted CSP 4, the phase is a chiral 2,2,2-trifluoro-1-(9-anthryl)ethanol tethered to silica by an elevencarbon-atom chain at the 10-position of the anthryl ring. CSP 4 typically provides greater enantioselectivity than its fluoroalcohol predecessor and is synthetically much more accessible.     

Monolithic silica columns with chemically bonded tert-butylcarbamoylquinine chiral anion-exchanger selector as a stationary phase for enantiomer separations

An enantioselective silica rod type chiral stationary phase (CSP) is presented as a novel combination of the well-known enantiomer separation properties of immobilized tert-butyl-carbamoylquinine chiral anion-exchanger selector with the unique properties of monolithic silica material. The chromatographic behavior of the tert-butyl-carbamoylquinine silica rod was studied and compared with a similar prepared particulate material. Good selectivities were achieved for a spectrum of chiral test components like N-derivatized amino acids (DNB- Ac-, DNZ-, Bz-, Z-amino acids) and for Suprofen. The influence of mobile phase parameters, as well as the effect of serially coupling up to six 10 cm monolithic silica columns was studied and put in context to conventional columns of particulate 5 microm type CSP. Using that 60 cm long monolithic column it was possible to improve the enantiomer separation of Suprofen and achieve a baseline separation in less than 10 min of total separation time.     

Unusually high enantioselectivity in high-performance liquid chromatography using cellulose tris(4-methylbenzoate) as a chiral stationary phase

The cellulose tris(4-methylbenzoate) chiral stationary phase (CSP) (commercially known as Chiralcel OJ-H) exhibited an extremely high enantioselectivity when used in the HPLC resolution of N-thiocarbamoyl-3-(4′-prenyloxy)-phenyl-5-phenyl-4,5-dihydro-(1H) pyrazole (Compound 1), in both normal-phase and polar organic conditions. Using n-hexane–ethanol (80:20, v/v) as a mobile phase, an enantioseparation factor value of 138.5 was found. In order to modulate the elution time of the longer retained enantiomer, a simple HPLC procedure was developed. The optimized analytical protocol was based on the stopped flow technique and did not involve any change in mobile phase composition. The stronger interaction energy of the (S) enantiomer compared to that of the (R) enantiomer was mainly attributed to the formation of a hydrogen bonding between the amino group of the thiocarbamoyl moiety and the carbonyl oxygen of the CSP.     

含磷手性化合物在多聚糖类手性固定相上的手性分离

在纤维素 三(3,5 二甲基苯基氨基甲酸酯)(ChiralcelOD)和直链淀粉 三(3,5 二甲基苯基氨基甲酸酯)(ChiralpakAD H)手性固定相上,采用高效液相色谱正相条件,分离了系列含磷手性化合物。考察了流动相中有机改性剂的种类及浓度对手性分离的影响;研究了化合物的结构与保留及对映体选择性的关系;并探讨了手性识别机理。     

The effect of analyte lipophilicity on the resolution of α-amino acids on a HPLC chiral stationary phase based on crown ether

The effect of analyte lipophilicity on the resolution of α-amino acids on a chiral stationary phase based on chiral crown ether has been examined by the chromatographic resolution trends for the resolution of a homologous series of five α-amino acids with an alkyl group of different length at the chiral center. The retention factors (k₁ and k₂) for the two enantiomers and the separation factors (α) were found to depend on the lipophilicity of the α-amino acid. In general, the retention factors increased as the organic modifier content in the mobile phase increased, the degree of the enhancement of retention factors being dependent on the analyte lipophilicity. The separation factors also increased as the analyte lipophilicity and the organic modifier content in the mobile phase increased. Possible rationales for these behaviors have been proposed.