Separation Of Enantiomers Research Articles

Study on Quality Control of Tenofovir Disoproxil Fumarate Enantiomers by High-Performance Liquid Chromatography–Mass Spectrometry

This study aims at developing a high-performance liquid chromatography–mass spectrometry (LC–MS) method to analyze tenofovir disoproxil fumarate (TDF) and its pharmaceutical preparations. Several cyclodextrin mobile-phase additives were applied to reversed-phase and normal-phase chromatography, and the effects of three chiral stationary phases on the TDF separation were investigated in this study. The R-type and S-type of TDF tablets were quantitatively analyzed in the single ion monitoring (SIM) scanning mode with a Unichiral CMD column. This method has been successfully applied to the separation and quantification of TDF and its isomers. The linear ranges of (R)-TDF and (S)-enantiomer were 1–20 and 0.2–16 μg/mL, respectively. The limit of detection for (R)-TDF and (S)-enantiomer was 0.0015 and 0.0012 μg/mL, respectively. (S)-enantiomer was not detected in the formulas from all the seven manufacturers, and the drug content of each took more than 98.5% of the labeled amount, which complies with the regulations. The method shows its advantages on high sensitivity, low detection limit, good practicability, and repeatability. The proposed method may provide a novel platform for separation of TDF enantiomers and quality control of TDF raw materials and preparations.

Chiral Separation of Cannabichromene, Cannabicyclol, and Their Acidic Analogs on Polysaccharide Chiral Stationary Phases.

Until recently, chirality has not been a major focus in the study of cannabinoids, as most cannabinoids of interest, such as cannabidiol and tetrahydrocannabinol, exist as a single isomer from natural sources. However, this is changing as more cannabinoids are identified, and compounds such as cannabichromene and cannabicyclol are emerging as potential investigatory candidates for varying indications. Because these molecules are chiral, the separation and study of the individual enantiomers' biological and physiological effects should therefore be of interest. The purpose of this study was to identify analytical separation conditions and then adapt those conditions to preparative separation. This was accomplished with a column-screening approach on Daicel's immobilized polysaccharide chiral stationary phases using non-traditional mobile phases, which included dichloromethane, ethyl acetate, and methyl tert-butyl ether under high-performance liquid chromatography conditions. CHIRALPAK® IK was found to separate all four compounds well with mobile phases containing hexane-dichloromethane (with or without an acidic additive). From these methods, the separation productivities were calculated to better visualize the separation scalability, which shows that the kilogram-scale separations of each are feasible.

NMR Discrimination of d- and l-α-Amino Acids at Submicromolar Concentration via Parahydrogen-Induced Hyperpolarization

Differentiation of enantiomers represents an important research area for pharmaceutical, chemical, and food industries. However, enantiomer separation is a laborious task that demands complex analytical techniques, specialized equipment, and expert personnel. In this respect, discrimination and quantification of d- and l-α-amino acids is no exception, generally requiring extensive sample manipulation, including isolation, functionalization, and chiral separation. This complex sample treatment results in high time costs and potential biases in the quantitative determination. Here, we present an approach based on the combination of non-hydrogenative parahydrogen-induced hyperpolarization and nuclear magnetic resonance that allows detection, discrimination, and quantification of d- and l-α-amino acids in complex mixtures such as biofluids and food extracts down to submicromolar concentrations. Importantly, this method can be directly applied to the system under investigation without any prior isolation, fractionation, or functionalization step.

Probing into complexation and separation of chiral‐at‐uranium complex to chiral sulphur enantiomers R/S‐ethiproles

Chiral pesticides R/S‐ethiproles(R/S‐EPLs) have different biotoxicity. Due to the difficulty in the separation of chiral sulphur enantiomers, it is of great significance to study the selective recognition and separation of chiral sulphur enantiomers R/S‐ethiproles. Based on DFT method, we theoretically designed a novel asymmetrical ligand of 4‐amino‐6‐(9‐[5‐hydroxy‐1H‐indol‐6‐yl]‐4,7‐dimethoxy‐1,10‐phenanthrolin‐2‐yl)‐1H‐indol‐5‐ol (AHPIL). Then we further constructed the chiral‐at‐uranium complex, namely, Uranyl‐AHPIL, to recognize and separate the enantiomers R/S‐EPLs. Various analyses, including electrostatic potential (ESP), molecular geometric structures, Mayer bond order (MBO), electron density difference diagram (EDDM), independent gradient model based on Hirschfield partition (IGMH), quantum theory of atoms in molecules (QTAIM), molecular orbitals, the extended transition state‐natural orbitals for chemical valence (ETS‐NOCV), hard–soft acid–base (HSAB), relaxed force constants (RFCs) and binding free energy, furthermore unravelled the selective complexation and chiral separation ability of Uranyl‐AHPIL receptor to R/S‐EPLs guests. The results all indicated that the novel ligand could coordinate with uranyl to form a stable chiral‐at‐uranium complex, which more inclined selectively to bind with R/S‐EPLs through thiyl oxygen O22, and its complexation ability to S‐EPL was much greater than to R‐EPL. In solvents water, acetone, dichloromethane, benzene, cyclohexane and in gas phase, the separation factors of Uranyl‐AHPIL towards R/S‐EPL were in range of 189–633, and the enantioselectivity coefficients were all above 99%. These results provide a theoretical guidance for the complexation and separation of chiral pesticides and at the same time offer a new reference for the design of actinide complexes.

Economic Separations of Organic Acidic or Basic Enantiomeric Mixtures—A Protocol Suggestion

In this review, we aim to present new concepts for the revisited separation of enantiomers from racemic compounds and a protocol worth to be followed in designing the preparation of pure enantiomers. We have taken into account not only the influence of the properties (eutectic composition) and characteristics of the reactants (racemic compound, resolving agent), but also the behavior of the resulting diastereomers and the different conditions (e.g., crystallization time, solvents used, solvate-forming compounds, achiral additives, etc.). The examples discussed are resolutions developed by our research team, through which we will try to illustrate the impact of all these considerations, presenting the methodological investigations interpreting recent discoveries and observations. Some special solid-state analytical and structural investigations assisting us in the elucidation and invention design of the resolution processes of some active pharmaceutical ingredients, such as Tetramisole, tofisopam, and Amlodipine, are also shown.

Carbon nanomaterials for designing next-generation membranes and their emerging applications.

Carbon nanomaterials have enormous applications in various fields, such as adsorption, membrane separation, catalysis, electronics, capacitors, batteries, and medical sciences. Owing to their exceptional properties, such as large specific surface area, carrier mobility, flexibility, electrical conductivity, and optical pellucidity, the family of carbon nanomaterials is considered as one of the most studied group of materials to date. They are abundantly used in membrane science for multiple applications, such as the separation of organics, enantiomeric separation, gas separation, biomolecule separation, heavy metal separation, and wastewater treatment. This study provides an overview of the significant studies on carbon nanomaterial-based membranes and their emerging applications in our membrane research journey. The types of carbon nanomaterials, their utilization in membrane-based separations, and the mechanism involved are summarized in this study. Techniques for the fabrication of different nanocomposite membranes are also highlighted. Lastly, we have provided an overview of the existing issues and future scopes of carbon nanomaterial-based membranes for technological perspectives.

Self-assembled stereomutation with supramolecular chirality inversion.

Supramolecular chirality is involved not only in biological events such as gene communication, replication, and enzyme catalysis but also in artificial self-assembly systems and aggregated materials. The precise control of supramolecular chirality, and especially supramolecular chirality inversion (SMCI), would deepen the understanding of chiral transfer and regulation in both living systems and artificial self-assembly systems, providing efficient ways to construct advanced chiral materials with an optimum assembly pathway necessary for various functions. In this review, the fundamental principles of SMCI are comprehensively summarized, with a focus on the helical assemblies having opposite handedness or chiroptical properties of the compositions. Thereafter, various SMCI strategies that have been developed for chiral nanostructures and assembled materials are systematically reviewed, and the promising applications of SMCI, including chiroptical switches, chiral recognition, enantiomeric separation, asymmetric catalysis, chiral optoelectronic materials, chiral spin filters, and biomedical uses, are highlighted accordingly. Finally, the scientific challenges and future perspectives for assembling materials with SMCI are also discussed.

Enantiomeric Separation by Capillary Electro chromatography with a Novel β-cyclodextrin-hyperbranched Carbosilane–silica Polymer Coating

A new type of β-cyclodextrin-modified hyperbranched carbosilane (β-CD@HBC) stationary phase was synthesized by combining hyperbranched carbosilane with the -OH group of β-cyclodextrin, and then the β-CD@HBC was used as astationary phase to fix it on the inner wall of capillary column, application in chiral drug separation. The chromatographic behaviour of the stationary phase was studied. The preliminary test results showed that the β-CD@HBC coated OT column has the ability to separate a variety of enantiomers, such as clenbuterol, ibuprofen, enrofloxacin, hlorpheniramine, isoproterenol hydrochloride, epinephrine, Phenylalanine, and Ketoprofen. The β-CD@HBC coated OT column has good repeatability and stability. The relative standard deviations (RSD) between runs, between days, and between columns were less than 1.5%, 1.86%, and 4.83%, respectively. This paper shows that hyperbranched polymers have great potential and advantages in the application of capillary electrochromatography for chiral separation.

Dielectric chiral metasurfaces for enhanced circular dichroism spectroscopy at near infrared regime.

Numerous applications of chiro-optical effects can be found in nanophotonics, including imaging and spin-selective absorption, particularly in sensing for separating and detecting chiral enantiomers. Flat single-layer metasurfaces composed of chiral or achiral sub-wavelength structures offer unique properties to manipulate the light due to their extraordinary light-matter interaction. However, at optical wavelengths, the generation of strong chirality is found to be challenging via conventional chiral metasurface approaches. This work intends to design and optimize a dielectric chiral meta-nano-surface based on a diatomic design strategy to comprehend giant chiro-optical effects in the near-infrared (NIR) regime for potential application in circular dichroism (CD) spectroscopy. Instead of using a single chiral structure that limits the CD value at optical wavelengths, the proposed metasurface used a diatomic (two meta-atoms with distinct geometric parameters) chiral structure as a building block to significantly enhance the chiro-optical effect. Combining both meta-atoms in a single periodicity of the building block introduces constructive and destructive interferences to attain the maximum circular dichroism value exceeding 75%. Moreover, using multipolar resonance theory, the physics behind the generation of giant chiro-optical effects have also been investigated. The proposed dielectric chiral meta-platform based on the extra degree of freedom can find application in compact integrated optical setups for CD spectroscopy, enantiomer separation and detection, spin-dependent color filters, and beam splitters.

Chiral probes for biosensing.

Chiral inorganic nanomaterials have emerged as a highly promising area of research in nanoscience due to their exceptional light-matter interaction and vast potential applications in chiral sensing, asymmetric catalysis, enantiomer separation, and negative-index materials. We present an overview of the latest advances in chiral inorganic nanomaterials including chiral individual nanoparticles, chiral assemblies, and chiral film-based sensors over the past ten years. Additionally, we discuss the challenges and future perspectives for developing chiral nanomaterials in biosensing applications.

Chiroptical regulation of macrocyclic arenes with flipping-induced inversion of planar chirality.

Studies on various macrocyclic arenes have received increasing attention due to their straightforward syntheses, convenient derivatization, and unique complexation properties. Represented by pillar[n]arenes, several distinctive macrocyclic arenes have recently emerged with the following characteristics: they possess a pair of enantiomeric planar chiral conformations, and interconversion between these enantiomeric conformations can be achieved through the flipping of ring units. Complexation of a chiral guest with these macrocyclic arenes will lead to a shift of the equilibrium between the Rp and Sp conformers, leading to intriguing possibilities for chiral induction and sensing. By the introduction of bulky substituents on the rims, employing rotaxanation or pseudocatenation, planar chirality could be locked, enabling the enantiomeric separation of the chiral structures. The induced or separated chiral conformers/compounds exhibit significant chiroptical properties. These macrocyclic arenes, with flipping-induced inversion of planar chirality, demonstrated intriguing chiral induction dynamics and kinetics. In this featured review, we systematically summarize the progress in chiroptical induction/regulation of these macrocyclic arenes, particularly in the fields of chiral sensing, molecular machines, molecular recognition, and assembly.

Novel approaches to the formation of coatings based on gold nanoparticles and albumin for chiral separation in capillary electrophoresis

Combination of properties of gold nanoparticles and bovine serum albumin is promising for the formation of chiral stationary phases enabling high enantioselectivity due to developed surface of coatings and increased chiral selector concentration on the capillary walls. In this work we proposed and compared two approaches to the formation of physically adsorbed multilayer coatings based on citrate-stabilized gold nanoparticles (cGNP) and bovine serum albumin (BSA) for the chiral separation in capillary electrophoresis. In the first approach pre-synthesized cGNP modified with BSA were immobilized on the capillary coated by poly(diallyldimethylammonium chloride) (PDADMAC). PDADMAC polymer was used as a binding layer promoting sorption of the nanoparticles on the capillary surface. It was shown that cGNP-BSA was poorly adsorbed on the capillary surface due to low zeta potential and could not be used for the formation of dense coatings. The second approach included sequential layer-by-layer deposition of PDAMAC and cGNP, resulting in the formation of a dense layer of nanoparticles. The main stage included in-capillary functionalization of cGNP with BSA. Scanning electron microscopy confirmed that the layer-by-layer deposition of modifiers ensured the formation of dense layer of nanoparticles on the capillary surface. The coating was stable in the entire range of pH studied (2–10). The application of such coating allowed reduction of BSA concentration (to 5 μM) in the background electrolyte required for the tryptophan enantiomers separation. It confirms the prospectiveness of combining nanoparticles and chiral selectors for the increase of specific surface area of the capillary inner walls.

Quantitative Analysis and Enantiomeric Separation of Ephedra Alkaloids in Ma Huang Related Products by HPLC-DAD and UPLC-MS/MS

Ephedra is a genus of the Ephedraceae family and is found in temperate regions, such as Central Asia and Europe. Among the various ephedra species, Ma Huang (Ephedra herb) is derived from the aerial parts of Ephedra sinica S tapf, Ephedra equisetina Bunge, and Ephedra intermedia Schrenk & C.A. Mey. Ma Huang contains various ephedra alkaloids, including (-)-ephedrine, (+)-pseudoephedrine, (-)-norephedrine, (+)-norpseudoephedrine, (-)-methylephedrine, and (+)-methylpseudoephedrine, which are found naturally as single enantiomers, although they can be prepared as racemates. Although the use of Ma Huang in foods is prohibited in Korea, products containing Ma Huang can be imported, and so it is necessary to develop a suitable analytical technique for the detection of Ma Huang in foods. Herein, we report the development of analytical methods for the detection of ephedra alkaloids in products containing Ma Huang. Following sample purification by solid phase extraction, quantitative analysis was performed using ultra-performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-MS/MS). Additionally, the enantiomers were successfully separated using HPLC-DAD. We successfully analyzed various food samples, where the ephedra alkaloids were qualitatively and quantitatively determined, and the enantiomers were separated. It is expected that these methods may contribute toward preventing the distribution of illegal products containing Ma Huang.

Selection and Analytical Performances of the Dragonfly Mass Spectrometer Gas Chromatographic Columns to Support the Search for Organic Molecules of Astrobiological Interest on Titan.

Titan is a key planetary body for astrobiology, with the presence of a subsurface ocean and a dense atmosphere, in which complex chemistry is known to occur. Approximately 1-Titan-year after the Cassini-Huygens mission arrived in the saturnian system, Dragonfly rotorcraft will land on Titan's surface by 2034 for an exhaustive geophysical and chemical investigation of the Shangri-La organic sand sea region. Among the four instruments onboard Dragonfly, the Dragonfly Mass Spectrometer (DraMS) is dedicated to analyze the chemical composition of surface samples and noble gases in the atmosphere. One of the DraMS analysis modes, the Gas Chromatograph-Mass Spectrometer (GC-MS), is devoted to the detection and identification of organic molecules that could be involved in the development of a prebiotic chemistry or even representative of traces of past or present life. Therefore, DraMS-GC subsystem should be optimized to detect and identify relevant organic compounds to meet this objective. This work is focused on the experimental methods employed to select the chromatographic column to be integrated in DraMS-GC, to assess the analytical performances of the column selected, and also to assess the performances of the second DraMS-GC column, which is devoted to the separation of organic enantiomers. Four different stationary phases have been tested to select the most relevant one for the separation of the targeted chemical species. The results show that the stationary phase composed of polymethyl (95%) diphenyl (5%) siloxane is the best compromise in terms of efficiency, robustness, and retention times of the molecules. The combination of the general and the chiral columns in DraMS is perfectly suited to in situ chemical analysis on Titan and for the detection of expected diverse and complex organic compounds.

Continuous flow synthesis and separation of mandelic acid enantiomers in a modular microfluidic system

Mandelic acid enantiomers are key precursors in the synthesis of valuable chiral products. However, separating enantiomers in reasonable amounts is a challenging task despite advances in chiral chromatography and other discontinuous and expensive techniques. To tackle these problems, we designed and tested a modular microfluidic system working in a fully continuous flow regime. The system consists of two modules, namely (i) a packed-bed microreactor (PBR) with immobilized enzyme lipase and (ii) a membrane microseparator driven by an imposed electric field. The immobilized catalyst converts a racemic substrate, methyl mandelate, into mandelic acid enantiomers. The enzyme preferentially synthesizes (R)-(−)-mandelic acid within minutes. The product stream from PBR is fed into a membrane microseparator operated in a counter-current regime. An inserted dialysis membrane and an orthogonally applied electric field completely separate the electrically charged enantiomers from their mixture with unreacted and uncharged methyl mandelate within 1.5 min. Under improved reaction and separation conditions, approximately 1 g of (R)-(−)-mandelic acid are produced per day with almost 60 % enantiomeric excess. Moreover, the productivity of this system is easily scalable without compromising its excellent reaction-transport characteristics.

Study of Different Chiral Columns for the Enantiomeric Separation of Azoles Using Supercritical Fluid Chromatography

The enantiomeric separation of antifungal compounds is an arduous task in pharmaceutical and biomedical fields due to the different properties that each diastereoisomer presents. The enantioseparation of a group of fungicides (sulconazole, bifonazole, triadimefon and triadimenol) using supercritical fluid chromatography was achieved in this work. For this goal, four different chiral columns based on polysaccharide derivatives, as well as the effect of different chromatographic parameters such as temperature, type and percentage of organic modifier (methanol, ethanol and isopropanol), were thoroughly investigated. The inversion of the elution order of enantiomers as a result of a change in the stationary phase or organic modifier was also evaluated by employing a circular dichroism detector. The best separation conditions, in terms of the enantioresolution and analysis time, were obtained with the Lux® Cellulose-2 column using isopropanol as the organic modifier.

Method Development, Optimization, and Validation of the Separation of Ketamine Enantiomers by Capillary Electrophoresis Using Design of Experiments

Capillary electrophoresis was chosen as cost-effective and robust method to separate ketamine enantiomers. For the method development, first different native and modified cyclodextrins were tested. The most promising chiral selector was α-cyclodextrin. A design of experiments (DoE) was carried out, which started with the screening of relevant factors. Based on these results, the method was optimized according to the significant factors (buffer, cyclodextrin concentration, pH value, voltage, temperature) of the screening based on the response resolution and migration time of the later migrating enantiomer. The optimized conditions consisted of a background electrolyte with 275 mM TRIS, adjusted with 85% phosphoric acid to a pH of 2.50, and 50 mM α-cyclodextrin, at a temperature of 15 °C, an applied voltage of 30 kV and an injection pressure of 1.0 psi for 10 s. A fused-silica capillary with a total length of 70 cm and an effective length to the detector of 60 cm was used. The method was validated according to ICH guideline Q2 R(1). The limit of quantification was 3.51 µg mL−1 for S-ketamine and 3.98 µg mL−1 for R-ketamine. The method showed good linearity for racemic ketamine with R2 of 0.9995 for S-ketamine and 0.9994 for R-ketamine. The lowest quantifiable content of S-ketamine found in R-ketamine was 0.45%.

Development of a capillary electrophoretic method for determination of ketorolac enantiomers in human plasma using cationic β-cyclodextrin derivative as a chiral selector.

A novel approach for the separation of ketorolac enantiomers by capillary electrophoresis is presented. A cationic β-cyclodextrin derivative based on imidazole was synthesized and used as a chiral selector in the background electrolyte. The influence of pH and ionic strength of background electrolyte, as well as cationic β-cyclodextrin derivative concentration on the resolution of ketorolac enantiomers, was investigated. The highest value of the resolution for ketorolac enantiomers was 1.46 when the background electrolyte consisted of 25mM NaH2 PO4 (pH 6.4) with 1mM 1-butyl-3-β-cyclodextrinimidazolium tosylate. Additionally, the possibilities of cationic derivatives for the separation of ketoprofen enantiomers were shown (peak resolution 1.06). The two-step preconcentration mode was developed to reduce the limit of detection of individual enantiomers. The proposed approach was successfully applied to determine ketorolac enantiomers in tablet "Ketorol express" and human plasma. The calibration range of ketorolac enantiomers for plasma samples was 0.25-2.50μg/ml with coefficients of determination ≥ 0.99. The relative standard deviation both of the peak area and migration time was less than 15%, as well as the accuracy ranged from 90.1% to 110.2% for both analytes. The limits of detection were 44 and 55ng/ml for R- and S-ketorolac. The quantity of ketorolac in plasma was verified with high-performance liquid chromatography.

Poly(carboxyethyl acrylate-co-ethylene glycol dimethacrylate) precursor monolith with bonded (S)-(-)-1-(2-naphthyl) ethylamine ligands for use in chiral and achiral separations by capillary electrochromatography

In this research report, the previously developed poly(carboxyethyl acrylate-co-ethylene glycol dimethacrylate) precursor monolith (referred to as carboxy monolith) is further exploited in the preparation of a chiral stationary phase for enantiomeric separations. The carboxy monolith precursor was subjected to post polymerization functionalization (PPF) with the chiral selector (S)-(-)-1-(2-naphthyl) ethylamine (NAS) at room temperature in the presence of N, N´-dicyclohexylcarbodiimide (DCC) in chloroform. The DCC, which is an organic soluble carbodiimide, permits the linkage for the amine functionality of the chiral ligand NAS to the carboxy group of the monolithic surface forming a stable amide linkage. The NAS column thus obtained allowed not only enantiomeric separations in the RP mode via its chiral site but also the separation of nonpolar species via its achiral functionality offering both hydrophobic and π-π interactions for aromatic compounds such toluene derivatives and polyaromatic hydrocarbons. The dual interaction sites (e.g., chiral, and achiral) of the NAS present a convenient column for the separations of slightly polar and nonpolar chiral and achiral solutes in the RP mode.

Structurally diverse isobutylamides from Zanthoxylum nitidum

Five pairs of new racemic alkamides (1a/1b and 4a/4b-7a/7b) and two new achiral derivatives (2–3), as well as five known ones (8–12), were purified from the 95% EtOH extract of Zanthoxylum nitidum. Their structures were elucidated based on spectroscopic analyses (NMR and HR-ESI-MS), electronic circular dichroism (ECD) and NMR calculations. The enantiomeric separation was successfully achieved by chiral-phase HPLC-ECD measurements. Among all the isolates, compounds 2, 3, and 10 showed inhibitory effects against five human cancer cell lines, with IC50 values in range of 18.51–48.03 μM.