The chromatographic behavior and thermodynamic characteristics of adsorption of profen enantiomers on silica gel with grafted eremomycin antibiotic

The chromatographic retention and thermodynamic characteristics of adsorption of several derivatives of 2-arylpropanoic acid on a chiral stationary phase with a grafted eremomycin antibiotic under the conditions of liquid chromatography with water-ethanol mobile phases were studied. The dependences of retention characteristics and selectivity on eluent pH were revealed. The difference between the adsorption mechanisms of acids with one and two benzene rings was demonstrated. Compensation effect manifestations in these systems are discussed.     

Liquid chromatographic separation of enantiomers using chiral additives in the mobile phase

Addition of an optically active compound to the mobile phase is an attractive method for resolving enantiomers in liquid chromatography. The technique is practical, easy to use and allows rapid screening for new chiral complexing agents as well as for optimal separation conditions.     

高效液相色谱手性流动相添加法拆分奥昔布宁对映体

采用C18固定相,以羟丙基-β-环糊精为手性流动相添加剂,建立了奥昔布宁对映体的高效液相色谱拆分方法。考察了手性添加剂、有机极性调节剂、缓冲盐的种类和浓度以及流动相的pH值和流速及柱温等因素对对映体分离的影响。在最佳分离条件下,奥昔布宁对映体的分离度为1.54,检测限为1.0 ng。该方法简便,重复性好,比手性固定相法更加经济。     

New chiral stationary phase with macrocyclic glycopeptide antibiotic eremomycin chemically bonded to silica

A new chiral stationary phase (CSP) was prepared by attachment of macrocyclic glycopeptide antibiotic eremomycin to the epoxy-activated silica under mild conditions. In contrast to CSP with immobilized vancomycin, which is a close structural analogue of eremomycin, the prepared CSP reveals high enantioselectivity for separation of amino acids enantiomers. It was demonstrated by the example of ristocetin A CSP that method of the immobilization of macrocyclic glycopeptide antibiotics affects remarkably the resulting enantioselectivity.     

Chromatographic resolution of tryptophan enantiomers with L-Leu-L-Leu-L-Leu peptide effects of mobile phase composition and chromatographic support

Tryptophan enantiomers have been separated by zwitterion pair chromatography using L-leucine-L-leucine-L-leucine peptide as the zwitterion pairing agent. The peptide ligand is adsorbed onto an octadecylsilane support with excess ligand present in bulk solution. This article examines the roles of the hydrophobic matrix and the mobile phase components on tryptophan enantiomer binding and resolution. Capacity factors and enantioselectivites are given for both hydrophobic and hydrophilic matrices using mobile phases containing Leu-Leu-Leu peptide and/or salt. A decrease in selectivity upon the addition of mobile phase salt suggests that quadrupolar ion-pairing contributes to chiral recognition. Results indicate that binding is significantly reduced and separation is not achieved when Leu-Leu-Leu is coupled onto cross-linked or polymerized hydrophilic resins as well as onto macroporous polystyrene resin. However, resin-immobilized Leu-Leu-Asp-Leu-Leu-Leu, Leu-Leu-Glu-Leu-Leu-Leu, and Leu-Leu-Leu-Glu-Leu-Leu peptides, with ion-pairing sites designed to mimic the Leu-Leu-Leu-saturated C18 support, also do not resolve tryptophan enantiomers. This suggests the Leu-Leu-Leu structure is critical for enantiomer resolution. Because D- and L-tryptophan are separated in the absence of bulk Leu-Leu-Leu, chiral discrimination is believed to occur at the surface of the octadecylsilane support.     

Semi-preparative gas chromatographic separation of all-trans-perhydrotriphenylene enantiomers on a chiral cyclodextrin stationary phase

Enantiomers of all-trans-perhydrotriphenylene (PHTP) were separated by gas chromatography using heptakis(6-O-tert.-butyldimethylsilyl-2,3-di-O-methyl)-beta-cyclodextrin (TBDMS-beta-CD) as the chiral selector. Conditions for semi-preparative separations were established using a 2 m x 2 mm I.D. packed column and subsequently extended to a 1.8 m x 4 mm I.D. column which enabled separations on a mg scale. The column packing was TBDMS-beta-CD dissolved in SE-54 coated on Chromosorb P AW-DMCS 80-100 mesh. Optimization of the chromatographic conditions (oven temperature, carrier gas flow, and column load) with respect to better efficiency and peak retention resulted in a system capable of separating up to 10 mg of the racemate per day. Purities of separated enantiomers were determined by capillary gas chromatography. Yields and purities of the fractions obtained by single- and double-step separations are compared. Highly enriched enantiomers with purities of up to 99.6% (99.2% ee) were obtained by a single separation step.     

Planar chromatographic separation of enantiomers and diastereomers with cyclodextrin mobile phase additives

A variety of racemic compounds were resolved using reversed-phase thin-layer chromatography (TLC) with mobile phases containing highly concentration solutions of beta-cyclodextrin (beta-CD). These include the drugs labetalol and mephenytoin, metallocenes, crown ethers, methyl-p-toluenesulfinate, nornicotine derivaties and several dansyl and beta-naphthylamide substituted amino acids. It was possible to resolve some racemates that could not be separated on beta-CD bonded phase liquid chromatography (LC) columns with this technique. Likewise there were some compounds that could be resolved with the LC approach that failed to separate with the present TLC method. In cases of racemates that could be resolved by either approach, it was found that the retention order was exactly opposite for the two methods. Enantiomeric resolution is highly dependent on mobile phase composition. In particular, the type and amount of organic modifier as well as the concentration of beta-CD affect the observed resolution. Possible reasons for the chromatographic behavior are discussed. Several diastereoisomeric compounds were separated as well, including steroid epimers and pharmaceutical compounds.     

Liquid chromatographic separation of enantiomers of beta-amino acids using a chiral stationary phase

The enantiomers of both alpha-substituted beta-alanines and beta-substituted beta-alanines may be chromatographically separated using silica-bonded chiral stationary phases derived from N-acetylated alpha-arylalkylamines. The amino acids are chromatographed as alkyl esters of N-3,5-dinitrobenzoyl derivatives; separability factors range from 1.11 to 1.65 for nine alpha-substituted beta-alanines and from 1.08 to 1.20 for nine beta-substituted beta-alanines. The enantiomers of beta-aminoisobutyrate and beta-leucine, chiral beta-amino acids occurring in animal tissues and physiological fluids, are among those resolved. The enantiomers of R,S-beta-aminoisobutyrate and several related alpha-alkyl-beta-alanines were prepared by chromatographic resolution of diastereomeric dipeptides.     

Effect of the mobile phase composition on the retention behaviour of diphenylsilica pre-coated plates

The retention of some rifamycins and steroids on diphenyl bonded pre-coated silica gel plates, in relation to the mobile phase used, was examined by thin-layer chromatography. Neat organic solvents, non-aqueous and aqueous binary mixtures were tested as eluents. By comparison of retention data for rifamycins and steroids, respectively, under non-aqueous and aqueous conditions, a dual retention mechanism on this diphenyl phase was found. Interactions with the residual silanol groups seemed to prevail when employing as mobile phase the more lipophilic solvents tested, such as chloroform or dichloromethane, whereas interactions with the aryl groups of the bonded phase prevailed when using high polarity alcohols or aqueous mixtures. As a consequence, by changing the mobile phase, a large variation in selectivity with a concomitant change in retention order of the test compounds was observed.     

Liquid chromatographic separation of the enantiomers of beta-amino acids on a ligand exchange chiral stationary phase

A liquid chromatographic ligand exchange chiral stationary phase (CSP) derived from (S)-leucinol was applied in the separation of the enantiomers of 12 beta-amino acids. The resolution was quite successful especially for the enantiomers of beta-amino acids containing aromatic functional group in the side chain. The chromatographic resolution behaviors were dependent on the organic modifier and Cu(II) concentration in aqueous mobile phase and the column temperature.     

Direct chromatographic resolution of carnitine and O-acylcarnitine enantiomers on a teicoplanin-bonded chiral stationary phase.

R-(-)-Carnitine (vitamin B(T)) plays an important role in human energy metabolism, by facilitating the transport of long-chained fatty acids across the mitochondrial membranes. Its (S)-enantiomer acts as a competitive inhibitor of carnitine acetyltransferase, causing depletion of the body R-(-)-carnitine stock. Consequently, the separation of carnitine enantiomers is very important both to study their biological activities and to control the enantiomeric purity of pharmaceutical formulations. In the present paper we describe an easy, fast and convenient procedure for the separation of the enantiomers of carnitine and O-acylcarnitines by enantioselective HPLC on a laboratory-made chiral column containing covalently bonded teicoplanin as selector. High enantioselectivity factors (alpha values ranging from 1.31 to 3.02) and short-time analyses characterize the analytical procedure; in addition, analytes are easily detected by evaporative light scattering with no need for preliminary derivatization. The effects of pH and ionic strength of the mobile phase and of the nature of the organic modifier on the enantioselective separations were also investigated.     

手性流动相添加剂法拆分盐酸氯丙那林对映体

盐酸氯丙那林(Clorprenaline hydrochloride)其化学名为α-[[(1-甲基乙基)氨基]甲基]-2-氯-苯甲醇盐酸盐,分子中含有一个手性碳原子,分子式见图1.临床上使用的是盐酸氯丙那林外消旋体,可用于支气管哮喘、哮喘型支气管炎、慢性支气管炎合并肺气肿,可止喘并改善肺功能.     

Liquid chromatography determination of citalopram enantiomers using beta-cyclodextrin as a chiral mobile phase additive

A reliable and specific method for the determination of citalopram enantiomers was developed and validated. Chromatographic resolution of citalopram enantiomers was made on a Shim-pack (5 microm particle size) cyanopropyl column with beta-cyclodextrin (beta-CD) as an effective chiral mobile phase additive. The composition of the mobile phase was (90 + 10, v/v) aqueous 0.1% triethylammonium acetate buffer, pH 4.0 (adjusted with acetic acid), and acetonitrile, containing 12 mM beta-CD. The flow rate was 0.8 mL/min with ultraviolet detection at 240 nm. The effects of the mobile phase composition, concentration of beta-CD, and pH of the triethylammonium acetate buffer on peak shape and resolution of the enantiomers were investigated. The calibration graphs were linear (r = 0.9999, n = 8) in the range of 1-40 microg/mL for S(+) citalopram and R-(-) citalopram. The limit of detection values were 5.51 x 10(-3) and 4.35 x 10(-3) pg/mL, while the limit of quantification values were found to be 1.84 x 10(-2) and 1.45 x 10(-2) microg/mL for S-(+) citalopram and R-(-) citalopram, respectively.     

高效液相色谱手性流动相添加剂法拆分氯噻酮对映体

应用反向高效液相色谱,以羟丙基-β-环糊精(HP--βCD)作为手性流动相添加剂,拆分了氯噻酮(Chlorthalidone)对映体。对主要的影响因素如羟丙基-β-环糊精浓度、流动相pH值、三乙胺(TEA)、甲醇、柱温、流速等进行了系统研究,建立了羟丙基-β-环糊精流动相添加剂法拆分氯噻酮对映体的方法。使用HarbonLichrospher-C18色谱柱(5μm,150 mm×4.6 mm),流动相为V(甲醇)∶V(水相)=20∶80(水相含30 mmol/L HP--βCD0、.1 mol/L Na2HPO4、体积分数2%的TEA、pH5),流速为0.8 mL/min,室温下拆分,氯噻酮对映体可得到良好分离。     

手性流动相添加法拆分酮康唑外消旋体

采用C18反相色谱柱,利用在流动相中加入手性选择剂的方法实现酮康唑对映体的拆分。研究了手性选择剂的种类及浓度、流动相pH值、甲醇比例和柱温等因素对酮康唑手性分离的影响,结果表明磺丁基-β-环糊精可以使酮康唑对映体完全分离,最后选择的流动相组成为甲醇-0.02 mol/L磷酸二氢钠(体积比为60∶40,含0.02%三乙胺和1.0 mmol/L磺丁基-β-环糊精,用稀磷酸调节pH值到3.00)。酮康唑对映体在6 min内得到基线分离,分离度为2.05。方法简便,分离效果好,对酮康唑对映体的拆分具有应用价值。     

Effect of mobile phase composition on the retention of selected alkaloids in reversed-phase liquid chromatography with chaotropic salts

Sodium hexafluorophosphate, perchlorate and trifluoroacetate were applied as ion-ion interaction reagents in reversed-phase liquid chromatography. The separation of chosen alkaloids was performed by changing the kind of the organic modifier (methanol, acetonitrile, tetrahydrofuran), concentration of the ion-ion-interaction reagents and the concentration of phosphate buffer at constant pH (2.7) in the mobile phase. Obtained results were analyzed in connection to a dynamic ion-exchange model of retention and ion-ion interaction effects. The perturbation method was applied to test proposed retention theories. The formation of ion-complexes controlling the retention in chaotropic systems was confirmed. On the basis of the relationships of capacity factors (k) versus salt concentrations derived experimentally, absolute increases in capacity factors, the desolvation parameters and the limiting retention factors were calculated and compared for all the investigated compounds in eluent systems studied. The selectivity of the proposed mobile phases was compared on the basis of the separation of alkaloid mixture.     

High performance liquid chromatographic separation of clausenamide enantiomers with chiral –AGP stationary phase

<正>A method of high performance liquid chromatographic separation of clausenamide enantiomers with chiral-AGP(α_1-acid glycoprotein) stationary phases has been established.The absolute configurations of(-)clausenamide and(+)clausenamide are 3S, 4R,5R,6S and 3R,4S,5S,6R,respectively.The present method has been used to analyze the(-)clausenamide and(+)clausenamide and its analogues such as the major metabolite and synthetic derivatives of clausenamide.     

Gas chromatographic resolution of carbohydrate enantiomers. A new chiral phase for pentoses

New chiral polymers have been prepared by conversion of cyano groups of polysiloxanes XE-60 and OV-225 into carboxylic groups and subsequent coupling to give the respective S-valine-R-α-phenyl-ethyl amide. In contrast to the corresponding stationary phases with S-α-phenylethyl amide groups, which have been used for the resoultion of trifluoroacetylated hexose derivatives, the new phase show high enantioselectivity for pentose and 6-deoxyhexose derivatives.