ABSTRACT
The present study investigates the utilization of a supramolecular deep eutectic solvent (SUPRADES), consisting of sulfated-ß-cyclodextrin (S-ß-CD) and citric acid (CA), as a chiral selector (CS) in capillary electrophoresis for the enantiomeric separation of nefopam (NEF) and five cathinone derivatives (3-methylmethcathinone [3-MMC], 4-methylmethcathinone [4-MMC], 3,4-dimethylmethcathinone [3,4-DMMC], 4-methylethcathinone [4-MEC], and 3,4-methylendioxycathinone [MDMC]). A significant improvement in enantiomeric separation of the target analytes was observed upon the addition of S-ß-CD-CA to the background electrolyte (BGE), leading to a baseline separation of all analytes. In particular, the optimum percentage of S-ß-CD-CA, added to the BGE, was determined to be 0.075% v/v for NEF (Rs = 1.5) and 0.050% v/v for three out of five cathinone derivatives (Rs = 1.5, 1.6, and 2.4 for 3-MMC, 4-MEC, and 3,4-DMMC, respectively). In the case of 4-MMC and MDMC, a higher percentage of the CS, equal to 0.075% and 0.10% v/v, respectively, was required to achieve baseline separation (Rs = 1.5, 1.9 for MDMC and 4-MMC, respectively). The outcomes of the present study highlight the potential effectiveness of using SUPRADES as a CS in electrophoretic enantioseparations.
ABSTRACT
In this study, an optimized environmentally friendly procedure was employed to enhance the sustainable utilization of phenolic antioxidants derived from aloe vera rind by-products. The procedure involved the application of ultrasound-assisted extraction (UAE) in combination with deep eutectic solvents (DESs). Eleven different DESs and three conventional solvents were employed as extraction media for polyphenolic compounds. Choline chloride-citric acid (ChCl-CA) was selected as the most suitable extractant, considering its extraction efficiency in relation to the total phenolic content. The operating conditions of UAE were optimized and modeled by the use of response surface methodology in order to maximize the yield of total phenolics and antioxidant capacity. The optimal operational parameters for the UAE procedure were determined to be 16.5 min, 74% (v/v) DES in water, and a solvent-to-solid ratio equal to 192. HPLC analysis, which was performed on the optimum extract, revealed significant levels of phenolics present in the aloe rind. Efficient recovery of the extracted antioxidants was obtained by the use of solid-phase extraction (SPE) and polyamide cartridges. The ChCl-CA DES exhibited excellent recycling capability with a yield of over 90% through SPE. Finally, the greenness of the method was evaluated using the green AGREE and AGREEprep metrics. The results highlighted the sustainability and the greenness of the proposed extraction procedure for the aloe by-product.
ABSTRACT
In this study, a novel supramolecular deep eutectic solvent consisting of sulfated-ß-CD and citric acid (S-ß-CD-CA) is reported for the first time. This innovative system was evaluated as a sole chiral selector in capillary electrophoresis for the enantioseparation of six fluorine-substituted amphetamine analogs, yielding remarkable outcomes. Baseline separations of all amphetamine analogs under study were achieved in less than 21.00 min using the S-ß-CD-CA as the chiral selector. It was observed that the addition of 0.050 % v/v S-ß-CD-CA into the background electrolyte resulted in the baseline separation of five out of the six fluorine-substituted amphetamine analogs, while in the case of the para-substituted amphetamine analog, 4-fluoramphetamine (4-FA), a higher percentage (0.15 % v/v) was required to achieve baseline enantioseparation. These findings emphasized the potential of this new supramolecular system in providing a class of solvents with promising chiral recognition properties.
Subject(s)
beta-Cyclodextrins , Amphetamine , Deep Eutectic Solvents , Fluorine , Solvents , Electrophoresis, Capillary/methods , Sulfates , StereoisomerismABSTRACT
A comprehensive study was performed to determine an optimum enantioseparation method for fluorine-substituted amphetamine and cathinone derivatives (fluor-amphetamine and fluor-cathinone derivatives), using a binary system consisting of carboxymethyl-ß-CD (CM-ß-CD) and a deep eutectic solvent (DES), namely choline chloride-ethylene glycol (ChCl-EG). Under this framework, the optimization and modeling of the separation conditions in a binary system were performed with the objective of maximizing resolution and minimizing analysis time. This was achieved through the application of response surface methodology. In particular, the effect of chiral selector concentration and percentage of DES on resolution and analysis time were investigated and optimized using a complete experimental design. The optimum enantioseparation conditions were determined to be 13.84 mM CM-ß-CD and 0.15% v/v ChCl-EG for fluorine-substituted amphetamine derivatives and 14.36 mM and 0.75% v/v ChCl-EG for fluorine-substituted cathinone derivatives, respectively. This combination resulted in a baseline separation for eight out of the nine analytes studied. Overall, the results demonstrated the synergistic effect of the CM-ß-CD/DES dual system and highlighted the significance of DESs as additives in capillary electrophoresis.
Subject(s)
Deep Eutectic Solvents , Fluorine , Electrophoresis, Capillary/methods , Choline , Amphetamines , StereoisomerismABSTRACT
The effect of the combined use of amino acid-based ionic liquids (AAILs) and deep eutectic solvents (DESs) with either cyclodextrin- (CD) or cyclofructan- (CF) based chiral selectors for the chiral separation of amphetamine derivatives was investigated in the present study. A non-significant improvement in enantiomeric separation of target analytes was observed when AAILs were combined with either CF or CD. On the other side, a markedly improved chiral separation of enantiomers was obtained using the dual carboxymethyl-ß-cyclodextrin/DES system, highlighting the existence of a synergistic effect. After the addition of 0.5% v/v of choline chloride-ethylene glycol, the resolution of the enantiomers of amphetamine, methamphetamine and 3-fluorethamphetamine, increased from 1.4, 1.1, 1.0 to 1.8, 1.8, and 1.5 min, and the analysis times increased from 19.54, 20.48, 18.71 to 35.71, 35.78 and 32.90 min, respectively. This was not the case for the CF/DES dual system, in which the separation of amphetamines worsened, indicating an antagonistic effect. In conclusion, DESs are a very promising additive in capillary electrophoresis that can improve the separation of chiral molecules in combination with CDs but not CFs.
Subject(s)
Cyclodextrins , Ionic Liquids , Amino Acids , Deep Eutectic Solvents , Electrophoresis, Capillary/methods , Stereoisomerism , AmphetamineABSTRACT
In recent years, carob and its derived products have gained wide attention due to their health-promoting effects, which are mainly attributed to their phenolic compounds. Carob samples (carob pulps, powders, and syrups) were analyzed to investigate their phenolic profile using high-performance liquid chromatography (HPLC), with gallic acid and rutin being the most abundant compounds. Moreover, the antioxidant capacity and total phenolic content of the samples were estimated through DPPH (IC50 98.83-488.47 mg extract/mL), FRAP (48.58-144.32 µmol TE/g product), and Folin-Ciocalteu (7.20-23.18 mg GAE/g product) spectrophotometric assays. The effect of thermal treatment and geographical origin of carobs and carob-derived products on their phenolic composition was assessed. Both factors significantly affect the concentrations of secondary metabolites and, therefore, samples' antioxidant activity (p-value < 10-7). The obtained results (antioxidant activity and phenolic profile) were evaluated via chemometrics, through a preliminary principal component analysis (PCA) and orthogonal partial least square-discriminant analysis (OPLS-DA). The OPLS-DA model performed satisfactorily, differentiating all samples according to their matrix. Our results indicate that polyphenols and antioxidant capacity can be chemical markers for the classification of carob and its derived products.
Subject(s)
Antioxidants , Fabaceae , Antioxidants/chemistry , Flavonoids/chemistry , Chemometrics , Phenols/chemistry , Fabaceae/chemistryABSTRACT
During this study, a simple and easy-to-prepare electrophoretic method was developed for the enantioseparation of amphetamine and cathinone derivatives. Different types of ß-cyclodextrin and cyclofructan-based chiral selectors (CSs), both native and derivatized, were utilized, and the most effective ones, in terms of resolution and analysis time, were identified. In addition, several electrophoretic parameters, such as background electrolyte concentration and pH, and CS concentration, were examined to optimize the separation conditions. Under the optimal electrophoretic conditions, 10 psychoactive substances were enantiomerically separated using 1 mM sulfated cyclofructan-6 (SCF-6) for the amphetamine derivatives and 1 mM sulfated cyclofructan-7 (SCF-7) for the cathinone derivatives dissolved in an aqueous solution of 20-mM monobasic sodium phosphate at pH 2.5, a temperature of 25°C, and an applied voltage of 25 kV. In addition, the method was validated by estimating the intra- and interday precision.
Subject(s)
Cyclodextrins , Cyclodextrins/chemistry , Stereoisomerism , Electrophoresis, Capillary/methods , Amphetamine , SulfatesABSTRACT
An ultrasound-assisted method was used for the extraction of phenolics from defatted black cumin seeds (Nigella sativa L.), and the effects of several extraction factors on the total phenolic content and DPPH radical scavenging activity were investigated. To improve the extraction efficiency of phenolics from black cumin seed by ultrasonic-assisted extraction, the optimal extraction conditions were determined as follows: ethanol concentration of 59.1%, extraction temperature of 44.6 °C and extraction time of 32.5 min. Under these conditions, the total phenolic content and DPPH radical scavenging activity increased by about 70% and 38%, respectively, compared with conventional extraction. Furthermore, a complementary quantitative analysis of individual phenolic compounds was carried out using the HPLC-UV technique. The phenolic composition revealed high amounts of epicatechin (1.88-2.37 mg/g) and rutin (0.96-1.21 mg/g) in the black cumin seed extracts. Ultrasonic-assisted extraction can be a useful extraction method for the recovery of polyphenols from defatted black cumin seeds.
Subject(s)
Catechin , Cuminum , Nigella sativa , Antioxidants/pharmacology , Catechin/analysis , Ethanol , Nigella sativa/chemistry , Phenols/chemistry , Plant Extracts/pharmacology , Polyphenols/analysis , Rutin/analysis , Seeds/chemistryABSTRACT
Extracts derived from the Ceratonia siliqua L. (carob) tree have been widely studied for their ability to prevent many diseases mainly due to the presence of polyphenolic compounds. In this study, we explored, for the first time, the anti-cancer properties of Cypriot carobs. We produced extracts from ripe and unripe whole carobs, pulp and seeds using solvents with different polarities. We measured the ability of the extracts to inhibit proliferation and induce apoptosis in cancer and normal immortalized breast cells, using the MTT assay, cell cycle analysis and Western Blotting. The extracts' total polyphenol content and anti-oxidant action was evaluated using the Folin-Ciocalteu method and the DPPH assay. Finally, we used LC-MS analysis to identify and quantify polyphenols in the most effective extracts. Our results demonstrate that the anti-proliferative capacity of carob extracts varied with the stage of carob maturity and the extraction solvent. The Diethyl-ether and Ethyl acetate extracts derived from the ripe whole fruit had high Myricetin content and also displayed specific activity against cancer cells. Their mechanism of action involved caspase-dependent and independent apoptosis. Our results indicate that extracts from Cypriot carobs may have potential uses in the development of nutritional supplements and pharmaceuticals.
Subject(s)
Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Fabaceae/chemistry , Phenols/chemistry , Phenols/pharmacology , Solvents/chemistry , Apoptosis/drug effects , Caspases/metabolism , Cell Line, Tumor , Fruit/chemistry , Humans , Seeds/chemistryABSTRACT
In this work, a simple, reliable, and fast capillary electrophoretic method was developed and validated for the simultaneous determination of 12 polyphenolic compounds, the most frequently found in carob's pulp and seeds. The present work deals with the development of a novel dual electrophoretic system based on the combined use of ß-CD and ionic liquid (IL) as buffer additives. A baseline separation of the target analytes was achieved in less than 10 min by using a BGE consisting of 35 mM borate along with 15 mM ß-CD and 3 mM l-alanine tert butyl ester lactate (l-AlaC4 Lac) IL as buffer additives at pH 9.5, a temperature of 25°C, and an applied voltage of 30 kV. The application of the developed electrophoretic method to real samples enabled the identification and quantification of the main phenolic constituents of both ripe and unripe carob pulp extracts. The results revealed the predominance of gallic acid in both ripe (183.92 µg/g carob pulp) and unripe (205.10 µg/g carob pulp) carob pulp and highlighted the great influence of the ripening stage on carobs polyphenolic composition, with unripe pods being more enriched in polyphenols (total phenolics detected: 912.58 and 283.13 µg/g unripe and ripe carob pulp).
Subject(s)
Fabaceae , Ionic Liquids , Phenols , beta-Cyclodextrins , Electrolytes , Galactans , Mannans , Plant GumsABSTRACT
Polyphenols in carobs have recently attracted great attention due to their wide range of biological and health promoting effects. A comprehensive study was conducted to find an optimum method for the extraction, purification and characterization of these valuable bioactive substances. Under this framework, the ultrasound-assisted extraction (UAE) of polyphenols from carob pulp was optimized by the maximization of the yield in total phenolics using response surface methodology. In particular, the effects of solid-solvent ratio, solvent concentration, extraction time, sonication amplitude, and sonication mode were investigated and optimized using a complete experimental design. In comparison to conventional extraction techniques, UAE offered a higher yield of antioxidants and a shorter processing time. Solid-phase extraction was evaluated as a clean-up strategy prior to the electrophoretic analysis of extracts. The results from the analysis of real samples revealed the predominance of gallic acid and highlighted the great influence of the ripening stage on carobs composition.
Subject(s)
Antioxidants/chemistry , Antioxidants/isolation & purification , Chemical Fractionation/methods , Galactans/chemistry , Mannans/chemistry , Plant Gums/chemistry , Polyphenols/analysis , Ultrasonic Waves , Solvents/chemistryABSTRACT
Cannabis is by far the most widely abused illicit drug globe wide. The analysis of its main psychoactive components in conventional and non-conventional biological matrices has recently gained a great attention in forensic toxicology. Literature states that its abuse causes neurocognitive impairment in the domains of attention and memory, possible macrostructural brain alterations and abnormalities of neural functioning. This suggests the necessity for the development of a sensitive and a reliable analytical method for the detection and quantification of cannabinoids in human biological specimens. In this review, we focus on a number of analytical methods that have, so far, been developed and validated, with particular attention to the new "golden standard" method of forensic analysis, liquid chromatography mass spectrometry or tandem mass spectrometry. In addition, this review provides an overview of the effective and selective methods used for the extraction and isolation of cannabinoids from (i) conventional matrices, such as blood, urine and oral fluid and (ii) alternative biological matrices, such as hair, cerumen and meconium.
Subject(s)
Cannabinoids/analysis , Chromatography, Liquid/methods , Mass Spectrometry , Substance Abuse Detection/methods , Body Fluids/chemistry , Cannabinoids/isolation & purification , Forensic Toxicology/methods , Hair/chemistry , Humans , Tandem Mass SpectrometryABSTRACT
The combined use of chiral ionic liquids (CILs) and conventional chiral selectors (CSs) in CE, to establish a synergistic system, has proven to be an effective approach for the separation of enantiomeric pairs. In this study, a new CE method was developed, employing a binary system of a CS, either a cyclodextrin (CD) or a cyclofructan (CF), and a chiral amino acid ester-based ionic liquid (AAIL), for the chiral separation of four basic, acidic and zwitterionic drug compounds. In particular, the enantioseparation of two anticoagulants, warfarin (WAR) and coumachlor (COU), a non-opioid analgesic, nefopam (NEF) and a third-generation antihistamine, fexofenadine (FXD), was examined, by supporting the BGE with a CS and the chiral AAIL L-alanine tert butyl ester lactate (L-AlaC4 Lac). Parameters, such as the type of the CS, the concentration of both the CS and L-AlaC4 Lac, and the BGE pH, were methodically examined in order to optimize the chiral separation of each analyte. It was observed that, in some cases, the addition of the AAIL into the BGE improved both resolution (Rs ) and efficiency (N) significantly. In other cases, the synergistic effect enabled baseline separation of analyte enantiomers, at a much lower concentration of the CS. Finally, after optimization of separation conditions, baseline separations (Rs >1.5) of all four analytes were achieved in less than 5 min.
Subject(s)
Cyclodextrins/chemistry , Electrophoresis, Capillary/methods , Fructans/chemistry , Ionic Liquids , Alanine/chemistry , Hydrogen-Ion Concentration , Ionic Liquids/chemistry , Ionic Liquids/isolation & purification , Lactates/chemistry , Pharmaceutical Preparations/chemistry , Pharmaceutical Preparations/isolation & purification , StereoisomerismABSTRACT
Carob (Ceratonia Siliqua L., tree of the pea family Fabaceae) and its products have recently attracted great interest due to their polyphenolic composition. This review summarizes the polyphenolic compounds that are contained in different carob parts (leaves, pod, seeds, barks) and products (syrup, flour, fiber). It also states the main differences of polyphenolic composition due to environmental and natural reasons, such as region, variety, and gender, and due to the processes used for preparation, extraction and analysis. The gender, along with the extraction procedure, proved to be the most important factors affecting the polyphenolic composition. Supercritical fluid extraction is the most efficient technique used because it protects polyphenols from decomposition. Due to the relatively low number of publications, it is concluded that further development of optimum methods for extraction, analysis and isolation of polyphenols should be carried over to assess their antioxidant capacity and their food technological and pharmaceutical industry.
Subject(s)
Fabaceae/chemistry , Food Technology , Polyphenols/analysis , Antioxidants/analysis , Dietary Fiber , SeedsABSTRACT
Accumulation of mechanical stresses during cancer progression can induce blood and lymphatic vessel compression, creating hypo-perfusion, hypoxia and interstitial hypertension which decrease the efficacy of chemo- and nanotherapies. Stress alleviation treatment has been recently proposed to reduce mechanical stresses in order to decompress tumor vessels and improve perfusion and chemotherapy. However, it remains unclear if it improves the efficacy of nanomedicines, which present numerous advantages over traditional chemotherapeutic drugs. Furthermore, we need to identify safe and well-tolerated pharmaceutical agents that reduce stress levels and may be added to cancer patients' treatment regimen. Here, we show mathematically and with a series of in vivo experiments that stress alleviation improves the delivery of drugs in a size-independent manner. Importantly, we propose the repurposing of tranilast, a clinically approved anti-fibrotic drug as stress-alleviating agent. Using two orthotopic mammary tumor models, we demonstrate that tranilast reduces mechanical stresses, decreases interstitial fluid pressure (IFP), improves tumor perfusion and significantly enhances the efficacy of different-sized drugs, doxorubicin, Abraxane and Doxil, by suppressing TGFß signaling and expression of extracellular matrix components. Our findings strongly suggest that repurposing tranilast could be directly used as a promising strategy to enhance, not only chemotherapy, but also the efficacy of cancer nanomedicine.
Subject(s)
Antineoplastic Agents/therapeutic use , Nanomedicine , Neoplasms/drug therapy , Particle Size , Stress, Mechanical , ortho-Aminobenzoates/therapeutic use , Animals , Antineoplastic Agents/pharmacology , Cell Line, Tumor , Collagen/metabolism , Down-Regulation/drug effects , Doxorubicin/pharmacology , Doxorubicin/therapeutic use , Drug Delivery Systems , Extracellular Fluid/metabolism , Extracellular Matrix/drug effects , Extracellular Matrix/metabolism , Gene Expression Regulation, Neoplastic/drug effects , Humans , Hyaluronic Acid/metabolism , Mice, Nude , Models, Biological , Neoplasms/blood supply , Neoplasms/genetics , Neoplasms/pathology , Perfusion , Signal Transduction/drug effects , Transforming Growth Factor beta/metabolism , Treatment Outcome , Tumor Microenvironment/drug effects , ortho-Aminobenzoates/pharmacologyABSTRACT
This report, which is a sequence of a series of reviews, records the most important chiral selectors (CSs) applied in CE. It highlights the CSs that were used during the period 2014 to mid-2016. In this review, method developments, validations, and pharmaceutical along with biomedical applications are presented. The different CSs include CDs, antibiotics, cyclofructants, linear and branched oligo- and polysaccharides, and polymeric surfactants. In addition, the advantages of these CSs, along with their chiral recognition mechanisms, and their performance, are discussed.
Subject(s)
Electrophoresis, Capillary/methods , Animals , Anti-Bacterial Agents/analysis , Cyclodextrins/analysis , Humans , Polysaccharides/analysis , Stereoisomerism , Surface-Active Agents/analysisABSTRACT
This review highlights recent advances and applications in open-tubular capillary electrochromatography (OT-CEC) for enantioseparations during the last decade. Although extensive research has been conducted in the area of separations by use of CEC, and a big number of reviews have been published, there is not a review on exclusively the use of chiral stationary phases (CSPs) in OT-CEC for enantioseparations. Therefore, in this review, the design and synthesis of different CSPs are presented, and their potential applications in OT-CEC for enantioseparations are discussed. The different approaches to CSP development include chiral nanomaterials, porous layers, molecular imprinting, sol-gel technique, polyelectrolyte multilayer coating, polymeric coating and others.
Subject(s)
Capillary Electrochromatography/trends , Polymers/chemistryABSTRACT
This paper reports the development and validation of a new method based on ultra-performance LC coupled to MS/MS for the simultaneous determination of four cholesterol oxidation products (COPs) in foodstuffs in only 4.1 min. The COPs were detected by ESI in positive-ion mode with multiple reaction monitoring, and the mass spectrometric conditions were optimized in order to increase sensitivity. The developed method was validated in terms of linearity, precision, LODs, and LOQs. Recoveries of the extraction process ranged from 86 to 98.5% when the samples were fortified at 100, 500, and 1500 ng/mL. The applicability of the method was confirmed by analyzing different food samples. Considering the paucity of data regarding the content of COPs in Cypriot foods, particular attention was devoted, for the first time, to the determination of the profile of the main COPs in widely consumed, traditional Cypriot foodstuffs (halloumi cheese, hiromeri, snails, etc.).
Subject(s)
Cholesterol/analysis , Chromatography, High Pressure Liquid/methods , Food Analysis/methods , Tandem Mass Spectrometry/methods , Animals , Cyprus , Dairy Products/analysis , Limit of Detection , Linear Models , Meat/analysis , Oxidation-Reduction , Rabbits , Reproducibility of Results , Snails , SwineABSTRACT
A fast screening method of whole blood was proposed for enantiorecognition of free L-T3 , L-T4 , and D-T4 . Stochastic microsensors based on four inulins (IN, IQ, TEX, and HD) immobilized on diamond paste (DP) were used for recognition of free L-T3 , L-T4 , and D-T4 . For the enantiorecognition of free L-T4 and D-T4 in whole blood and pharmaceutical samples, the best microsensor was the one based on TEX/DP (wide linear concentration ranges, and low limits of quantification). The best limit of detection for the assay of free L-T3 (400 fmol/L) was recorded using the microsensors based on HD/DP, while for the assay of free L-T4, and D-T4 the best limit of determination (1 pmol/L) was recorded using the TX/DP-based microsensor. For the enantiorecognition of free L-T3 in whole blood and pharmaceutical samples the best microsensor was the one based on HD/DP (the wider linear concentration range, and the lower limit of quantification - of pmol/L magnitude order). For the enantiorecognition of free L-T3 in whole blood and pharmaceutical samples the best microsensor was the one based on HD/DP (the wider linear concentration range, and the lower limit of quantification - of pmol/L magnitude order). Free L-T3 , L-T4 , and D-T4 were recovered with high reliabilities in whole blood samples (recoveries higher than 99.00%, with RSD values lower than 1.00%) and pharmaceutical samples (recoveries higher than 95.00% with RSD values lower than 1.00%).
Subject(s)
Thyroxine/blood , Triiodothyronine/blood , Humans , Limit of Detection , StereoisomerismABSTRACT
Cyclofructans (CFs) and their derivatives have recently been proven to be efficient chiral selectors (CSs) for the enantioseparation of several analytes in CE, HPLC, and GC. In this study, the chiral separation ability of a number of native and derivatized CFs was examined in CE. Particularly, six different CFs, with different derivatization groups and cavity sizes [native CF-6 and CF-7, isopropyl cyclofructan-6 (IPCF-6), IPCF-7, sulfated cyclofructan-6 (SCF-6), and SCF-7] were used as CSs for the enantioseparation of huperzine A, warfarin, and coumachlor. Almost all of the examined CFs, except from SCF-6 & -7, demonstrated relatively low and sometimes no chiral separation ability for huperzine A. In an effort to improve both resolution and efficiency, the chiral ionic liquid D-Alanine tert butyl ester lactate (D-AlaC4Lac) was added into the BGE. In most of the cases, the combination of CF with D-AlaC4Lac resulted in an improvement in peak efficiency and/or resolution. When CF-6 was utilized with D-AlaC4Lac, a resolution of 1.4 was obtained, while the use of IPCF-6/D-AlaC4Lac provided a baseline enantioseparation. Although the combination of SCF-7 and 40 mM D-AlaC4Lac did not affect resolution, it dramatically increased peak efficiency from 24,000 to 117,000. In the case of warfarin and coumachlor, IPCF-6 and IPCF-7 proved to be the most effective CSs. It is, therefore, concluded that the size of the cavity and the CF derivatization are the key parameters for the chiral separation capability. It is also clear from this study that D-AlaC4Lac is necessary for improved peak efficiencies and resolutions.