Separation of the four pairs of enantiomers of vincamine alkaloids by enantioselective high-performance liquid chromatography

The four enantiomeric pairs of vincamine group alkaloids were separated by HPLC using Chiralpak AD as chiral stationary phase (CSP) and various n-hexane-2-propanol and n-hexane-ethanol mobile phases. (+)-cis-Vincamine, which is used in pharmaceutical preparations, is eluted much faster than its optical isomer, with separation factors of 2.4 and 3.5, respectively in these mobile phases. Other CSPs gave negative results. A chiral recognition mechanism is proposed and circular dichroism spectra of the individual enantiomers are presented.     

Enantiomer separation of pharmaceuticals by capillary gas chromatography with novel chiral stationary phases

New chiral stationary phases of polydimethylsiloxane anchored with (S)-(-)-t-leucine derivatives were provided for use in enantiomer separation of pharmaceuticals by capillary gas chromatography. Fifteen pharmaceuticals were separated into their enantiomeric pairs by converting them into pentafluoropropionyl and heptafluorobutyryl derivatives. The separation factor and resolution obtained from the new phases were superior to those from the conventional Chirasil-Val capillary column.     

Comparison of HPLC enantioseparation of substituted binaphthyls on CD‐, polysaccharide‐ and synthetic polymer‐based chiral stationary phases

Retention and enantioseparation behavior of ten 2,2′‐disubstituted or 2,3,2′‐trisubstituted 1,1′‐binaphthyls and 8,3′‐disubstituted 1,2′‐binaphthyls, which are used as catalysts in asymmetric synthesis, was investigated on eight chiral stationary phases (CSPs) based on β‐CD, polysaccharides (tris(3,5‐dimethylphenylcarbamate) cellulose or amylose CSPs) and new synthetic polymers (trans‐1,2‐diamino‐cyclohexane, trans‐1,2‐diphenylethylenediamine and trans‐9,10‐dihydro‐9,10‐ethanoanthracene‐(11S,12S)‐11,12‐dicarboxylic acid CSPs). Normal‐, reversed‐phase and polar‐organic separation modes were employed. The effect of the mobile phase composition was examined. The enantiomeric separation of binaphthyl derivatives, which possess quite similar structures, was possible in different enantioselective environments. The substituents and their positions on the binaphthyl skeleton affect their properties and, as a consequence, the separation system suitable for their enantioseparation. In general, the presence of ionizable groups on the binaphthyl skeleton, substitution with non‐identical groups and a chiral axis in the 1,2′ position had the greatest impact on the enantiomeric discrimination. The 8,3′‐disubstituted 1,2′‐binaphthyl derivatives were the most easily separated compounds in several separation systems. From all the chiral stationary phases tested, cellulose‐based columns were shown to be the most convenient for enantioseparation of the studied analytes. However, the polymeric CSPs with their complementary behavior provided good enantioselective environments for some derivatives that could be hardly separated in any other chromatographic system.     

高效液相色谱法手性分离二茂铁衍生物

建立了4个单手性和3个双手性(含有手性中心和面手性)的二茂铁衍生物在Chiralpak IC(纤维素-三(3,5-二氯苯基氨基甲酸酯))和Chiralpak IE3(直链淀粉-三(3,5-二氯苯基氨基甲酸酯))手性固定相上的高效液相色谱分离方法。4个单手性二茂铁衍生物中有3个可以在Chiralpak IE3固定相上实现基线分离,另外1个则在Chiralpak IC手性固定相上实现基线分离。3个双手性二茂铁衍生物可在Chiralpak IC手性固定相上实现基线分离。研究表明,这两种手性固定相对二茂铁衍生物具有较好的手性识别作用,并且具有互补作用。这一研究结果可为手性二茂铁化合物的分离提供借鉴和参考。     

Asymmetric activation of chiral alkoxyzinc catalysts by chiral nitrogen activators for dialkylzinc addition to aldehydes: super high-throughput screening of combinatorial libraries of chiral ligands and activators by HPLC-CD/UV and HPLC-OR/RIU systems

Asymmetric catalysts, prepared by chiral ligand exchange or chiral modification, can evolve further into highly activated catalysts through engineering with chiral activators. Two new methodologies for "super high-throughput screening" (SHTS) of chiral ligands and activators have been developed as a combination of HPLC-CD/UV (CD/ UV = circular dichroism/ultraviolet spectroscopy) or -OR/RIU (OR/RIU = optical rotation/refractive index unit) with a combinatorial chemistry (CC) factory. With these techniques, the % ee of the product is determined within minutes without separation of the enantiomeric products by using a nonchiral stationary phase. Therefore, those SHTS techniques combined with our 'asymmetric activation' concept can provide a powerful strategy for finding the best activated chiral catalyst.     

Synthesis and chiral recognition of novel amylose derivatives containing regioselectively benzoate and phenylcarbamate groups

A new class of regioselectively substituted amylose derivatives bearing three different substituents at 2-, 3- and 6-positions, and two different substituents at 2-position and 3-, 6-positions were synthesized by a sequential process based on the esterification of 2-position of a glucose unit. Their chiral recognition abilities were evaluated as chiral stationary phases (CSPs) for high-performance liquid chromatography (HPLC). Each derivative had its own characteristic recognition ability depending on the arrangement of side chains at the three positions. Among the derivatives, amylose 2-(4-t-butylbenzoate) and amylose 2-(4-chlorobenzoate) series exhibited high chiral recognition. Some racemates can be efficiently separated on these derivatives as well as on the amylose tris-3,5-dimethylphenylcarbamate, which is commercially available as Chiralpak AD and one of the most powerful CSPs. The structures of the amylose derivatives were also investigated by circular dichroism spectroscopy.     

Electrostatic molecular potential contour maps generated from ab initioMODPOT/VRDDO/MERGE wave functions of carcinogenic 3-methylcholanthrene and its metabolites

The electrostatic molecular potential contour maps were calculated for carcinogenic 3-methylcholanthrene (3-MCA) and a number of its metabolites {3-MCA 7,8-oxide and 3-MCA 9,10-oxide; 3-MCA 7,8-dihydrodiols[several stereoisomers: A trans(equatorial, equatorial) and A cis(equatorial, axial)]; 3-MCA 9,10-dihydrodiol–7,8-epoxide A βββ and 3-MCA 9,10-dihydrodiol–7,8-epoxide A αβα}. The maps were generated from our ab initio MODPOT/VRDDO/MERGE wave functions calculated for these species. The results of these maps for 3-MCA [similarly to our results for the maps we generated for benzo(a)pyrene (BP)] show that these electrostatic molecular potential contour maps can be used to indicate favored positions of attack for electrophilic species, such as “electrophilic” oxygen to form an epoxide as well as for positive ion attack. The 3-MCA maps indicate the favored site for attack and the pathways. The maps around 3-MCA 9,10-oxide and around 3-MCA 9,10-dihydrodiol-7,8-epoxide indicate the directional preferences for proton assisted epoxide ring opening. The maps around the 3-MCA dihydrodiols indicate that while for certain stereoisomers the “electrophilic” oxygen will prefer to attack from below, for other isomers it will prefer to attack from above. This gives great insight into the stereochemical preference for formation of different 3-MCA 9,10-dihydrodiol–7,8-epoxides.     

Liquid chromatography of jasmonic acid amine conjugates

Summary Racemic jasmonic acid (3R,7R/3S,7S)-(±)-JA) was chemically conjugated with different biogenic amines originating from aliphatic and aromatic α-amino acids by decarboxylation. The resulting isomeric compounds were subjected to reversed-phase high-performance liquid chromatography (HPLC) and to HPLC on the chiral stationary phases Chiralpak AS and Nucleodex β-PM. Under reversed-phase conditions, all the homologous amine derivatives tested could be separated from each other except the JA-conjugates containing 2-phenyl-ethylamine and 3-methylbutylamine. On both chiral supports the (3R,7R)-(−)-JA conjugates eluted earlier than those of the enantiomeric counterpart (3S,7S)-(+)-JA. On Chiralpak AS all the isomers studied could be separated to baseline with a mobile phase containingn-hexane and 2-propanol. The calculated resolution factors were between 1.80 and 4.17. The pairs of isomers were also chromatographed on the cyclodextrin stationary phase Nucleodex β-PM with methanol-triethylammonium acetate buffer as mobile phase. Under these conditions resolution factors were between 0.74 and 1.29. The individual isomers were chiroptically characterized by measurement of their circular dichroism.     

HPLC semi‐preparative separation of diclazuril enantiomers and racemization in solution

Diclazuril has been widely used in poultry feed for prevention and treatment of coccidiosis, and its chiral separation is rarely reported. Herein, semi‐preparative separation method of diclazuril enantiomers has been developed through normal‐phase high‐performance liquid chromatography. Effects of chiral stationary phases, alcoholic modifiers, and column temperature on separation of diclazuril were discussed in detail. Both the single‐urea‐bound 4‐chlorophenylcarbamoylated β‐cyclodextrin and amylose tris(3,5‐dimethylphenylcarbamate)‐coated chiral stationary phases showed strong ability in separation of diclazuril by using n‐hexane–trifluoroacetic acid–ethanol. Then, semi‐preparative separation of diclazuril was carried out through stacked injection, and the "enantiomeric excess" purities of two fractions were over 98%. Next, the electronic circular dichroism profiles of these two fractions in ethanol solution displayed the mirror image of each other in the range 360–200 nm. Moreover, effects of acidic/basic additive, time, and temperature on racemization of diclazuril enantiomers in ethanol solution have been studied in detail through normal‐phase high‐performance liquid chromatography. Racemization of diclazuril enantiomers was remarkably accelerated through adding triethylamine at high temperature. We envision that this systematic investigation of diclazuril at an enantiomeric level would provide valuable information in future studies involving enantioselective bioactive, metabolic, and toxicological activities.     

Chiral separation of lenalidomide by liquid chromatography on polysaccharide‐type stationary phases and by capillary electrophoresis using cyclodextrin selectors

Complementary techniques were applied for the investigation of the chiral recognition and enantiomeric resolution of lenalidomide using various cyclodextrins and polysaccharides as chiral selectors. The high‐performance liquid chromatography enantioseparation of the anticancer drug was achieved using polysaccharide‐type chiral stationary phases in polar organic mode. Elution order and absolute configuration were elucidated by combined circular dichroism spectroscopy and time‐dependent density functional theory calculations after the isolation of pure enantiomers. Chiral selector dependent and mobile‐phase dependent reversal of the enantiomer elution order was observed, and the nonracemic nature of the lenalidomide sample was also demonstrated. Eight anionic cyclodextrins were screened for their ability to discriminate between the uncharged enantiomers by using capillary electrophoresis. Only two derivatives presented chiral interactions, these cases being interpreted in terms of apparent stability constants and complex mobilities. The best results were delivered by sulfobutylether‐β‐cyclodextrin, where quasi‐equal stability constants were recorded and the enantiodiscrimination process was mainly driven by different mobilities of the transient diastereomeric complexes. The optimized high‐performance liquid chromatography (Chiralcel OJ column, pure ethanol with 0.6 mL/min flow rate, 40°C) and capillary electrophoresis methods (30 mM sulfobutylether‐β‐cyclodextrin, 30 mM phosphate pH 6.5, 12 kV applied voltage, 10°C) were validated for the determination of 0.1% (R)‐lenalidomide as a chiral impurity, which could be important if a racemic switch is achieved.     

Separation of chiral furan derivatives by liquid chromatography using cyclodextrin-based chiral stationary phases

The enantiomeric separation of a set of 30 new chiral furan derivatives has been achieved on native and derivatized beta-cyclodextrin stationary phases using high performance liquid chromatography (HPLC). The hydroxypropyl-beta-cyclodextrin (Cyclobond RSP), the 2,3-dimethyl-beta-cyclodextrin (Cyclobond DM), and the acetyl-beta-cyclodextrin (Cyclobond AC) stationary phases are the most effective chiral stationary phases (CSPs) for the separation of these racemates in the reverse phase mode. No enantioseparations have been observed on the native beta-cyclodextrin chiral stationary phase (Cyclobond I 2000) and only a few separations have been attained on the S-naphthylethyl carbamate beta-cyclodextrin (Cyclobond SN) and 3,5-dimethylphenyl carbamate beta-cyclodextrin (Cyclobond DMP) chiral stationary phases in the reverse phase mode. The polar organic and the normal phase mode on these CSPs are not effective for separation of these compounds. The characteristics of the analytes, including steric bulk, hydrogen bonding ability, and geometry, play an important role in the chiral recognition process. The pH affects the enantioseparation of compounds with ionizable groups and the addition of 0.5% methyl tert-butyl ether to the mobile phase significantly enhances the separation efficiency for some highly retained compounds.     

Chiral HPLC separation and absolute configuration assignment of a series of new triazole compounds

A series of novel chiral triazole compounds were synthesized. They were separated into enantiomers by liquid chromatography on an amylose tris(3,5-dimethylphenylcarbamate) (ADMPC) chiral stationary phase (CSP). The absolute configuration of each enantiomer of the investigated compounds was established by combined use of chemical correlation, chiral HPLC and circular dichroism (CD) spectra analysis methods. The influence of the mobile-phase modifiers and the structure of chiral triazole compounds on the chiral separation and retention were investigated. Reversal of the elution order of some enantiomeric pairs upon using different mobile-phase modifier was observed. The temperature effect on the chiral separation and the thermodynamic properties including enthalpy and entropy change of binding to the ADMPC-CSP were also investigated.     

Enantiomeric impurities in chiral catalysts,auxiliaries, and synthons used in enantioselective syntheses. Part 4

The enantiomeric purity of chiral reagents used in asymmetric syntheses directly affects the apparent reaction selectivity and the product’s enantiomeric excess. Herein, 46 recently available chiral compounds were evaluated in order to determine their actual enantiomeric compositions. They have not been assayed previously and/or have been introduced after 2006, when the last comprehensive evaluation of commercially available chiral compounds was reported. These compounds are widely used in asymmetric syntheses as chiral synthons, catalysts, and auxiliaries. The enantioselective analysis methods include HPLC approaches using Chirobiotic, Cyclobond and LARIHC series chiral stationary phases, and GC approaches using Chiraldex chiral stationary phases. Accurate, efficient assays for selected compounds are given. All enantiomeric test results were categorized within five impurity levels (i.e., <0.01%, 0.01–0.1%, 0.1–1%, 1–10% and >10%). Different batches of the same reagent from the same company can have different levels of enantiomeric impurities. Many of the reagents tested were found to have less than 0.1% enantiomeric impurities. Only one of the chiral compounds was found to have an enantiomeric impurity exceeding 10%.     

Chiral stationary phases based on chitosan bis(4‐methylphenylcarbamate)‐(alkoxyformamide)

Highly N‐deacetylated chitosan was chosen as a natural chiral origin for the synthesis of the selectors of chiral stationary phases. Therefore, chitosan was firstly acylated by various alkyl chloroformates yielding chitosan alkoxyformamides, and then these resulting products were further derivatized with 4‐methylphenyl isocyanate to afford chitosan bis(4‐methylphenylcarbamate)‐(alkoxyformamide). A series of chiral stationary phases was prepared by coating these derivatives on 3‐aminopropyl silica gel. The content of the derivatives on the chiral stationary phases was nearly 20% by weight. The chiral stationary phases prepared from chitosan bis(4‐methylphenylcarbamate)‐(ethoxyformamide) and chitosan bis(4‐methylphenylcarbamate)‐(isopropoxyformamide) comparatively showed better enantioseparation capability than those prepared from chitosan bis(4‐methylphenylcarbamate)‐(n‐pentoxyformamide) and chitosan bis(4‐methylphenylcarbamate)‐(benzoxyformamide). The tolerance against organic solvents of the chiral stationary phase of chitosan bis(4‐methylphenylcarbamate)‐(ethoxyformamide) was investigated, and the results revealed that this phase can work in 100% ethyl acetate and 100% chloroform mobile phases. Because as‐synthesized chiral selectors did not dissolve in many common organic solvents, the corresponding chiral stationary phases can be utilized in a wider range of mobile phases in comparison with conventional coating type chiral stationary phases of cellulose and amylose derivatives.     

高效液相色谱-圆二色检测法分析甲霜灵的对映体纯度

以手性农药甲霜灵为研究对象,使用非手性高效液相色谱在不拆分对映体的条件下,利用圆二色检测器所测的各向异性系数(g)测定手性对映体纯度。实验结果表明,g与对映体过剩率(ee)具有良好的线性关系;通过比较g所测ee与手性色谱所测的ee,二者所测ee相对平均偏差小于3.0%,说明该方法具有较高的准确性,可应用于手性化合物对映体纯度的测定。     

Chromatographic evaluation of poly (trans-1,2-cyclohexanediyl-bis acrylamide) as a chiral stationary phase for HPLC

Chiral stationary phases (CSPs) based on polymeric (R,R)- or (S,S)-1,2-diaminocyclohexane (DACH) derivatives are synthesized. When bonded to 5 microm porous spherical silica gel, the poly (trans-1,2-cyclohexanediyl-bis acrylamide) based poly-cyclic amine polymer (P-CAP) stationary phases is proved to be effective chiral stationary phases that could be used in the normal-phase mode, polar organic mode and with halogenated solvents mobile phases, if desired. Since these are entirely synthetic CSPs, the elution order of all enantiomers can be reversed between the (R,R) P-CAP and (S,S) P-CAP columns. Because of the high loading of chiral selectors, the columns exhibit very high sample capacities. Thus, P-CAP columns are useful for preparative and semi-preparative enantiomeric separations. The application of these CSPs and optimization of their separations are discussed.     

Chiral liquid chromatography contribution to the determination of the absolute configuration of enantiomers

The review covers examples in which chiral HPLC, as a source of pure enantiomers, has been combined with classical methods (X-ray, vibrational circular dichroism (VCD), enzymatic resolutions, nuclear magnetic resonance (NMR) techniques, optical rotation, circular dichroism (CD)) for the on- or off-line determination of absolute configuration of enantiomers. Furthermore, it is outlined that chiral HPLC, which associates enantioseparation process and classical purification process, opens new perspectives in the classical determination of absolute configuration by chemical correlation or chemical interconversion methods. The review also contains a discussion about the various approaches to predict the absolute configuration from the retention behavior of the enantiomers on chiral stationary phases (CSPs). Some examples illustrate the advantages and limitations of molecular modeling methods and the use of chiral recognition models. The assumptions underlying some of these methods are critically analyzed and some possible emerging new strategies are outlined.     

Enantiomeric separation of chiral pesticides by high-performance liquid chromatography on an amylose tris-(S)-1-phenylethylcarbamate chiral stationary phase

Amylose tris-(S)-1-phenylethylcarbamate chiral stationary phase (CSP) was prepared. The direct enantiomeric separation of chiral pesticides on this CSP had been studied by HPLC. The mobile phase was n-hexane-isopropanol at a flow rate of 1.0 mL/min. The effects of isopropanol content and column temperature on retention and enantioselectivity were investigated. Thirty-two samples were tested, of which ten interacted enantioselectively with the CSP. Five samples were completely resolved and another five underwent near-baseline or partial resolution. The enantiomers were identified by a circular dichroism detector. Linear van't Hoff plots were established and the thermodynamic parameters were thus calculated.     

Enantioselective strong cation-exchange molecular recognition materials: design of novel chiral stationary phases and their application for enantioseparation of chiral bases by nonaqueous capillary electrochromatography

New chiral stationary phases derived from enantiomerically pure derivatives of cysteine carrying sulfonic acid groups are synthesized and evaluated for enantiomer separation of chiral bases by non aqueous capillary electrochromatography after bonding to a linker and grafting upon thiol-modified silica particles. Structural modifications of these low molecular weight chiral selectors are investigated and discussed in terms of apparent enantioselectivities and resolution factors based on the enantiomeric separations of a set of chiral bases including beta-blockers, beta-sympathomimetics and other basic drugs. The influence of the mobile phase constitution and its flow velocity on the enantioseparation by nonaqueous capillary electrochromatography is also briefly evaluated and discussed for the chiral substances investigated.     

Site-selective DNA alkylation of GG steps by naphthaldiimide derivatives possessing enantiomeric epoxide

We have synthesized an enantiomeric pair of novel DNA alkylating agents consisting of a naphthaldiimide intercalator and a chiral epoxy side chain. These naphthaldiimide derivatives have high DNA binding affinity and selectively alkylate 5'G of the GG steps for (S)-epoxide and 3'G for (R)-epoxide.