Cyclodextrin-modified Micellar Electrokinetic Chromatography Research Articles

Simultaneous Separation and Determination of Nine Active Ingredients in Sanyetangzhiqing by Cyclodextrin-Modified Micellar Electrokinetic Capillary Electrophoresis-Diode Array Detector.

A simple and sensitive strategy using cyclodextrin-modified micellar electrokinetic chromatography with diode array detector was developed and applied for the simultaneous separation and determination of nine components in Sanyetangzhiqing (SYTZQ), a hypoglycemic and hypolipidemic agent. Several important parameters affecting separation performance were evaluated and optimized using single variable methods. Under the optimal conditions, baseline separation of the nine components, including four flavonoids (hyperoside, isoquercitrin, quercetin-3-O-glucuronoside, and astragalin), four phenolic acids (chlorogenic acid, rosmarinic acid, salvianolic acid B, and lithospermic acid), and a monoterpenoids (paeoniflorin), were achieved in less than 16 min. The correlation coefficients of the calibration curves were over 0.9996 for all the analytes. Intraday and interday precisions ranged from 0.4% to 4.8% and 1.7% to 5.0%, respectively. Recoveries of analytes varied from 95.3% to 105%. Validation results as well as the application to analyse SYTZQ samples demonstrated the applicability of the proposed method and thus provided an effective tool for the quality control of SYTZQ. Moreover, with the advantages of short time consuming, low energy consumption, high efficiency, and low cost, this method has laid a foundation for the determination and quality evaluation of multicomponents in Chinese herbal compounds.

Analytical Quality by Design-Compliant Development of a Cyclodextrin-Modified Micellar ElectroKinetic Chromatography Method for the Determination of Trimecaine and Its Impurities.

In 2022, the International Council for Harmonisation released draft guidelines Q2(R2) and Q14, intending to specify the development and validation activities that should be carried out during the lifespan of an analytical technique addressed to assess the quality of medicinal products. In the present study, these recommendations were implemented in Capillary Electrophoresis method development for the quality control of a drug product containing trimecaine, by applying Analytical Quality by Design. According to the Analytical Target Profile, the procedure should be able to simultaneously quantify trimecaine and its four impurities, with specified analytical performances. The selected operative mode was Micellar ElectroKinetic Chromatography employing sodium dodecyl sulfate micelles supplemented with dimethyl-β-cyclodextrin, in a phosphate-borate buffer. The Knowledge Space was investigated through a screening matrix encompassing the composition of the background electrolyte and the instrumental settings. The Critical Method Attributes were identified as analysis time, efficiency, and critical resolution values. Response Surface Methodology and Monte Carlo Simulations allowed the definition of the Method Operable Design Region: 21-26 mM phosphate-borate buffer pH 9.50-9.77; 65.0 mM sodium dodecyl sulfate; 0.25-1.29% v/v n-butanol; 21-26 mM dimethyl-β-cyclodextrin; temperature, 22 °C; voltage, 23-29 kV. The method was validated and applied to ampoules drug products.

Enantioseparation of Selected Imidazole Drugs Using Dual Cyclodextrin-Modified Micellar Electrokinetic Chromatography.

Particular attention has been paid to capillary electrophoresis as versatile and environmentally friendly approach for enantioseparations of a wide spectrum of compounds. Cyclodextrin-modified micellar electrokinetic chromatography (CD-MEKC) is a method of choice to provide effective separation toward hydrophobic and uncharged stereoisomers. The chiral discrimination of the solutes relies upon the partitioning between a given CD in the aqueous phase and micelles formed from a surfactant. Synergistic combinations of chiral selectors, surfactant, and modifier contribute to successful enantioseparations of the enantiomers. In this chapter, an application of CD-MEKC for the enantioseparation of selected imidazole drugs employing a dual CDs system is described.

Combining excitation-emission matrix fluorescence spectroscopy, parallel factor analysis, cyclodextrin-modified micellar electrokinetic chromatography and partial least squares class-modelling for green tea characterization

In this study, an alternative analytical approach for analyzing and characterizing green tea (GT) samples is proposed, based on the combination of excitation–emission matrix (EEM) fluorescence spectroscopy and multivariate chemometric techniques. The three-dimensional spectra of 63 GT samples were recorded using a Perkin–Elmer LS55 luminescence spectrometer; emission spectra were recorded between 295 and 800 nm at excitation wavelength ranging from 200 to 290 nm, with excitation and emission slits both set at 10 nm. The excitation and emission profiles of two factors were obtained using Parallel Factor Analysis (PARAFAC) as a 3-way decomposition method. In this way, for the first time, the spectra of two main fluorophores in green teas have been found. Moreover, a cyclodextrin-modified micellar electrokinetic chromatography method was employed to quantify the most represented catechins and methylxanthines in a subset of 24 GT samples in order to obtain complementary information on the geographical origin of tea. The discrimination ability between the two types of tea has been shown by a Partial Least Squares Class-Modelling performed on the electrokinetic chromatography data, being the sensitivity and specificity of the class model built for the Japanese GT samples 98.70% and 98.68%, respectively. This comprehensive work demonstrates the capability of the combination of EEM fluorescence spectroscopy and PARAFAC model for characterizing, differentiating and analyzing GT samples.

Chiral analysis of theanine and catechin in characterization of green tea by cyclodextrin-modified micellar electrokinetic chromatography and high performance liquid chromatography

Monomeric catechins are important compounds in green tea accounting for potential bioactivity against a wide range of diseases. Besides catechins, l-Theanine (γ-glutamylethylamide), a characteristic amino acid in tea leaves, has become a further focus of the phytochemical research for the reported beneficial effects mainly on cognitive performance, emotional state and sleep quality. In the present study has been developed a CD-MEKC method based on sodium dodecyl sulfate (SDS) and Heptakis (2,6-di-O-methyl)-β-cyclodextrin for the separation of six major green tea catechins and enantiomers of theanine. The latter, because of the poor detectability was derivatized prior analysis by o-phthaldialdehyde in the presence of N-acetyl-l-cysteine which, under mild conditions (neutral pH, in two minutes) allowed two diastereomers isoindole derivatives to be obtained. The derivatization reaction was directly carried out on tea infusion and derivatized samples were analysed by CD-MEKC involving 65 mM SDS and 28 mM cyclodextrin in acidic buffer (pH 2.5). The separation of six major green tea catechins including enantioresolution of (±)-Catechin and d/l-Theanine was obtained in about 5 min allowing d-Theanine to be quantified at least at 0.5% m/m level with respect to l-Theanine. Since (−)-Catechin and d-Theanine can be considered as non-native enantiomers (distomers), their presence in real samples provides an indication of tea leaves treatments (thermal treatment, fermentation, etc.) and could represent an opportunity for grading tea. The obtained results were confirmed by a RP-HPLC approach; even though the chromatography was developed in achiral conditions, the derivatization approach applied to theanine (diastereomers formation), allowed for d/l-Theanine chiral analysis.

Determination of Saikosaponins in Bupleuri Radix by Micellar Electrokinetic Chromatography with Experimental Design

ABSTRACTThe important pharmacological components of Bupleuri radix include saikosaponins a–d. A multivariable approach with experimental design was employed in this study to determine four saikosaponins in different species of Bupleuri radix by cyclodextrin-modified micellar electrokinetic chromatography. Compared to traditional multiple-parameter investigations, the current experimental design can shorten the time necessary for method development, and the multivariable approach can be used to evaluate the separation capability of the method and to explore the interactions of the different parameters. The significant influences of two experimental variables, including the concentration of γ-cyclodextrin and the percentage of methanol, on the responses were investigated using chromatographic resolution statistics. The optimized conditions were as follows: 25 mM of borate (pH 9.18) containing 50 mM of sodium dodecyl sulfate, 5 mM of γ-cyclodextrin, and 15% methanol (v/v) as running buffer. After the separation conditions were optimized and good resolution was acquired, solid phase extraction was performed to reduce the interference of crude extracts of Bupleurum kaoi and Bupleurum chinense DC. The simple and rapid analytical method was successfully applied, allowing for the determination of four saikosaponins and the identification of species of Bupleuri radix.

Melatonin in Edible and Non-Edible Plants

The concept of melatonin has become more important recently both in plants and in human who utilize plants for nutritional and health purposes. Melatonin, synthesized from L-tryptophan by enzyms, protects plants against difficult conditions. People have consumed these plants for their antioxidant, immunomodulator, antiinflammatory and anticancer effects. In parts of edible and non-edible plants, levels of melatonin are determined by cyclodextrin-modified micellar electrokinetic chromatography, enzyme-linked immuno sorbent assay, radioimmunoassay, high-performance liquid chromatography, liquid chromatography with electrochemical detection, liquid chromatography with fluorimetric detection, liquid chromatography-mass spectrometry, and liquid chromatography-ultraviolet spectrophotometry. In this review, biosynthesis of melatonin in both animal and plants, function of melatonin in plant kingdom, especially in medicinal/edible and nonedible plants, and detection of phytomelatonin content in those plants are presented.

Approaching over 10000-fold sensitivity increase in chiral capillary electrophoresis: Cation-selective exhaustive injection and sweeping cyclodextrin-modified micellar electrokinetic chromatography.

A novel and simple method that combines an online concentration technique with an enantioseparation technique for capillary electrophoresis-namely, cation-selective exhaustive injection and sweeping cyclodextrin-modified micellar electrokinetic chromatography (CSEI-sweeping CD-modified MEKC)-realizes the effective enantioseparation of cationic analytes while keeping a significant increase of detection sensitivity. This technique consists of a slight modification of the basic CSEI-sweeping MEKC. The main idea is to simply add an anionic CD as a chiral selector into the micellar buffer including sodium dodecyl sulfate, but not to change any other buffers in order to preserve the online concentration mechanism. When applied to analysis of the street drug, methamphetamine, the method achieved not only a baseline enantioseparation but also limits of detection (LODs; S/N = 3) of 70-90 pg/mL (ppt) for each isomer. This translates to a more than 10000-fold improvement compared to the LODs by the usual injection method. The present technique, which was made from a slight modification of CSEI-sweeping MEKC, would give an attractive approach that is applicable to almost any analytes for which CSEI-sweeping MEKC is applicable; all that is required is the selection of an appropriate anionic CD to be added to the micellar buffer.

Enantioseparation and impurity determination of ambrisentan using cyclodextrin-modified micellar electrokinetic chromatography: Visualizing the design space within quality by design framework

A capillary electrophoresis method for the simultaneous determination of the enantiomeric purity and of impurities of the chiral drug ambrisentan has been developed following the Quality by Design principles. The selected separation system consisted of a micellar pseudostationary phase made by sodium dodecyl sulphate with the addition of γ-cyclodextrin. The effects of critical process parameters (capillary length, temperature, voltage, borate concentration, pH, sodium dodecyl sulphate concentration, γ-cyclodextrin concentration) on enantioresolution of ambrisentan and analysis time were extensively investigated by multivariate strategies involving a screening phase and Response Surface Methodology. The Design Space was defined with a desired probability level π≥90%, and the working conditions, with the limits of the Design Space, corresponded to the following: capillary length, 64.5cm; temperature, 22°C; voltage, 30kV (26–30kV); background electrolyte, 100mM borate buffer pH 9.20 (8.80–9.60), 100mM sodium dodecyl sulphate, 50mM (43–50mM) γ-cyclodextrin. A Plackett-Burman design was applied for robustness testing, and a method control strategy was established. The method was fully validated according to the International Conference on Harmonisation guidelines and was applied to ambrisentan coated tablets.

Chiral cyclodextrin-modified micellar electrokinetic chromatography and chemometric techniques for green tea samples origin discrimination

Catechins and methylxanthines were determined in 92 green tea (GT) samples originating from Japan and China by using micellar electrokinetic chromatography with the addition of (2-hydroxypropyl)-β-cyclodextrin. GT samples showed high concentrations of (−)-epigallocatechin gallate and caffeine, with (−)-epigallocatechin, (−)-epicatechin gallate and (−)-epicatechin in relevant content and (+)-catechin, (−)-catechin and theobromine in much lower amounts. The amount of all the considered compounds was higher for Chinese GTs, with the exception of (−)-epicatechin gallate. Pattern recognition methods were applied to discriminate GTs according to geographical origin, which is an important factor to determine quality and reputation of a commercial tea product. Data analysis was performed by principal component analysis and hierarchical cluster analysis as exploratory techniques. Linear discriminant analysis and quadratic discriminant analysis were utilized as discrimination techniques, obtaining a very good rate of correct classification and prediction.

Enantioseparation of enilconazole using cyclodextrin-modified MEKC

Enantiomeric separation of enilconazole has been successfully achieved using cyclodextrin modified micellar electrokinetic chromatography (CD-MEKC) method with HP-γ- CD as chiral selector. Two parameters which were HP-γ-CD concentration and SDS concentration have been optimized by investigating their effects on resolution and analysis time. Enantioresolution of enilconazole was achieved by the optimized CD-MEKC system containing 40 mM HP-γ- CD, 40 mM SDS and 20 mM phosphate buffer (pH 8) with resolution (Rs = 1.3) and analysis time within 9 min. Good repeatabilities were obtained in term of RSD% (n = 3) for migration time, peak area and peak height ranging from 0.07 to 6.1 %. The optimized CD-MEKC method was a simple, less solvent used, and able to perform enantiomeric separation of enilconazole with a good resolution and relatively short analysis time.

Electronic nose and chiral-capillary electrophoresis in evaluation of the quality changes in commercial green tea leaves during a long-term storage

Electronic nose and capillary electrophoresis were applied in quality control of green tea samples subjected to long-term storage. Twelve representative green teas were considered, available as an “aged” (tea leaves stored during a long-term period of two years) and/or “not aged” (fresh products) samples. Their infusions were analyzed by an electronic nose, equipped with an array of six metal oxide semiconductor (MOS) sensors to obtain olfactive fingerprints of the volatile compounds in the infusions headspace. Upon training and chemometric analysis of acquired data (linear discriminant analysis), the electronic nose was found to be able in correctly classifying unknown samples as “aged” or “not aged”. Concomitantly, the infusion samples were analyzed by Cyclodextrin-modified Micellar Electrokinetic Chromatography (CD-MEKC) for determination of catechins. The analysis of seven most represented catechins and the methylxanthines theobromine and caffeine revealed a general loss of the polyphenols in each of the considered aged samples (up to 45%, w/w). In addition, the applied enantioselective method based on (2-hydroxypropyl)-β-cyclodextrin (HP-βCD) as chiral selector, was exploited for the estimation of (+)-Gallocatechin in the presence of (−)-Gallocatechin; the latter, as the non-native enantiomer, can be associated to the epimerisation of (−)-Epigallocatechin and was assumed as a marker occurring in case of uncorrected storage conditions of tea leaves. Interestingly, it was observed that epimerization did not significantly occur during aging. The application of CD-MEKC and electronic nose allowed for a fast characterization of green teas taking into account that the aroma is a decisive parameter for the acceptance of the product, whereas the catechins content is associated to the biological value.

Robust analysis of the hydrophobic basic analytes loratadine and desloratadine in pharmaceutical preparations and biological fluids by sweeping—cyclodextrin-modified micellar electrokinetic chromatography

The analysis of hydrophobic basic analytes by micellar electrokinetic chromatography (MEKC) is usually challenging because of the tendency of these analytes to be adsorbed onto the inner capillary wall in addition to the difficulty to separate these compounds as they exhibit extremely high retention factors. A robust and reliable method for the simultaneous determination of loratadine (LOR) and its major metabolite desloratadine (DSL) is developed based on cyclodextrin-modified micellar electrokinetic chromatography (CD-MEKC) with acidic sample matrix and basic background electrolyte (BGE). The influence of the sample matrix on the reachable focusing efficiency is studied. It is shown that the application of a low pH sample solution mitigates problems associated with the low solubility of the hydrophobic basic analytes in aqueous solution while having advantages with regard to on-line focusing. Moreover, the use of a basic BGE reduces the adsorption of these analytes in the separation compartment. The separation of the studied analytes is achieved in less than 7min using a BGE consisting of 10mmolL−1 disodium tetraborate buffer, pH 9.30 containing 40mmolL−1 SDS and 20mmolL−1 hydroxypropyl-β-CD while the sample solution is composed of 10mmolL−1 phosphoric acid, pH 2.15. A full validation study of the developed method based on the pharmacopeial guidelines is performed. The method is successfully applied to the analysis of the studied drugs in tablets without interference of tablet additives as well as the analysis of spiked human urine without any sample pretreatment. Furthermore, DSL can be detected as an impurity in LOR bulk powder at the stated pharmacopeial limit (0.1%, w/w). The selectivity of the developed method allows the analysis of LOR and DSL in combination with the co-formulated drug pseudoephedrine. It is shown that in CD-MEKC with basic BGE, solute–wall interactions are effectively suppressed allowing the development of efficient and precise methods for the determination of hydrophobic basic analytes, whereas the use of a low pH sample solution has a positive impact on the attainable sweeping efficiency without compromising peak shape and resolution.

Determination of Three Phthalate Esters in Environmental Samples by Coal Cinder Extraction and Cyclodextrin Modified Micellar Electrokinetic Chromatography

A new micellar electrokinetic chromatography (MEKC) method using beta-cyclodextrin (ß-CD) as the electrophoresis additive has been developed for the simultaneous determination of dimethyl phthalate (DMP), diethyl phthalate (DEP) and di(2-ethylhexyl) phthalate (DEHP) in environmental samples. To improve the sensitivity of cyclodextrin-modified MEKC (CD-MEKC), a flow injection procedure using a microcolumn packed with coal cinder as the solid-phase extractant was also investigated for the preconcentration and separation of DMP, DEP and DEHP in environmental samples. Parameters affecting CD-MEKC separation and coal cinder flow injection solid-phase extraction were systematically researched. In the presence of the running buffer [5 mmol/L of borax, 5% (v/v) methanol and 25 mmol/L of sodium dodecyl sulfate at pH 9.5], the addition of 14 mmol/L ß-CD greatly improved the separation efficiency. The analytes were quantitatively adsorbed by coal cinders and readily desorbed quantitatively with 0.2 mL of 10% (v/v) methanol-10 mmol/L disodium hydrogen phosphate. Under the optimum conditions, the enrichment factor of coal cinder was 60, and the determination limits of DMP, DEP and DEHP were 3.07, 2.07 and 4.06 ng/mL, respectively. The presented procedure was successfully applied to determine DMP, DEP and DEHP in landfill leachate and water samples with satisfactory results.

Separation of Selected Imidazole Enantiomers Using Dual Cyclodextrin System in Micellar Electrokinetic Chromatography

Cyclodextrin-modified micellar electrokinetic chromatography (CD-MEKC) method was developed for simultaneous enantioseparation of three imidazole drugs namely tioconazole, isoconazole and fenticonazole. Three easily available and inexpensive cyclodextrins namely 2-hydroxypropyl-β-cyclodextrin (HP-β-CD), 2-hydroxypropyl-γ-cyclodextrin (HP-γ-CD) and heptakis(2,6-di-O-methyl)-β-cyclodextrin (DM-β-CD) were evaluated to discriminate the six stereoisomers of the drugs. However, none of the three CDs gave a complete enantioseparation of the drugs. Effective enantioseparation of tioconazole, isoconazole and fenticonazole was achieved using a combination of 35 mM HP-γ-CD and 10 mM DM-β-CD as chiral selectors. The best separation using both HP-γ-CD and DM-β-CD (35 mM:10 mM) as chiral selectors were accomplished in background electrolyte (BGE) containing 35 mM phosphate buffer (pH 7.0), 50 mM sodium dodecyl sulfate (SDS) and 15% (v/v) acetonitrile at 27 kV and 30 °C with all peaks resolved in less than 15 min with resolutions, Rs 1.90-27.22 and peak efficiencies, N > 180 000. The developed method was linear over the concentration range of 25-200 mg l(-1) (r(2) > 0.998) and the detection limits (S/N = 3) of the three imidazole drugs were found to be 2.7-7.7 mg l(-1). The CD-MEKC method was successfully applied to the determination of the three imidazole drugs in spiked human urine sample and commercial cream formulation of tioconazole and isoconazole with good recovery (93.6-106.2%) and good RSDs ranging from 2.30-6.8%.

Separation of phthalates by cyclodextrin modified micellar electrokinetic chromatography: Quantitation in perfumes

A new CE method has been developed for the simultaneous separation of a group of parent phthalates. Due to the neutral character of these compounds, the addition of several bile salts as surfactants (sodium cholate (SC), sodium deoxycholate (SDC), sodium taurodeoxycholate (STDC), sodium taurocholate (STC)) to the separation buffer was explored showing the high potential of SDC as pseudostationary phase. However, the resolution of all the phthalates was not achieved when employing only this bile salt as additive, being necessary the addition of neutral cyclodextrins (CD) and organic modifiers to the separation media. The optimized cyclodextrin modified micellar electrokinetic chromatography (CD-MEKC) method consisted of the employ of a background electrolyte (BGE) containing 25mM β-CD-100mM SDC in a 100mM borate buffer (pH 8.5) with a 10% (v/v) of acetonitrile, employing a voltage of 30kV and a temperature of 25°C. This separation medium enabled the total resolution of eight compounds and the partial resolution of two of the analytes, di-n-octyl phthalate (DNOP) and diethyl hexyl phthalate (DEHP) (Rs~0.8), in only 12min. The analytical characteristics of the developed method were studied showing their suitability for the determination of these compounds in commercial perfumes. In all the analyzed perfumes the most common phthalate was diethyl phthalate (DEP) that appeared in ten of the fifteen analyzed products. Also dimethyl phthalate (DMP), diallyl phthalate (DAP), dicyclohexyl phthalate (DCP), and di-n-pentyl phthalate (DNPP) were found in some of the analyzed samples.

Chemometrics-guided development of a cyclodextrin-modified micellar electrokinetic chromatography method with head-column field amplified sample stacking for the analysis of 5-lipoxygenase metabolites

A new sensitive method using α-cyclodextrin-modified micellar electrokinetic chromatography has been developed to separate and quantify arachidonic acid metabolites of the lipoxygenase pathways in human polymorphonuclear leukocytes, i.e. leukotriene B4, 6-trans-leukotriene B4, 6-trans-12-epi-leukotriene B4, 5(S)-hydroxy-6-trans-8,11,14-cis-eicosatetraenoic acid, 12(S)-hydroxy-6-trans-8,11,14-cis-eicosatetraenoic acid, and 15(S)-hydroxy-6-trans-8,11,14-cis-eicosatetraenoic acid. The electrophoresis system was optimized with regard to the pH, boric acid, SDS and α-cyclodextrin concentration as well as separation voltage and temperature using a three level resolution IV fractional factorial design and a five level circumscribed central composite design. The resulting optimized conditions included 80mM sodium borate buffer, pH 10.07, containing 16.6mM sodium dodecyl sulfate, and 15mM α-cyclodextrin, using a separation voltage of 12.5kV at 23°C. Sensitivity was enhanced employing head-column field amplified sample stacking which resulted in limits of quantification between 30 and 50ng/mL and limits of detection between 10 and 17ng/mL after solid phase extraction of the lipoxygenase products. The method was validated according to the recommendations of the International Conference on Harmonization and applied to the determination of the lipoxygenase metabolites in polymorphonuclear leukocytes upon stimulation with Ca2+-ionophore A23187 and arachidonic acid. Robustness was confirmed using a three level resolution IV fractional factorial design. The novel method is suitable for the analysis of various arachidonic acid metabolites produced by cells and may be used for evaluation of lipoxygenase inhibitors.

Analysis of Cyclosporin A and Main Degradation Impurities by Cyclodextrin–Modified Micellar Electrokinetic Chromatography

Separation of the immunosuppressive drug cyclosporin A (CyA) from the closely related degradation impurities cyclosporin H (CyH, a CyA diastereomer) and isocyclosporin A (IsoCyA) was accomplished by means of cyclodextrin-modified micellar electrokinetic chromatography (CD-MEKC). Heptakis (2,3,6-tri-O-methyl)-β-cyclodextrin (TM-βCD) showed to be an effective modifiers of the sodium dodecyl sulfate (SDS) micellar system, allowing for the CD-MEKC separation of CyA from CyH and IsoCyA. By means of electric current measurements carried out using the capillary electrophoretic apparatus, the critical micelle concentration of SDS in the presence of different neutral cyclodextrins was estimated. Interestingly, it was found that TM-βCD strongly inhibited the micellization of SDS compared to β-, and hydroxypropyl-β-cyclodextrin. This unusual behavior was considered to be a key factor of the specific ability of the methylated cyclodextrin in providing CD-MEKC separation of hydrophobic cyclosporins. The optimized method consisted of: 50 mM sodium dodecyl sulfate (SDS) in 50 mM tetraborate buffer (pH 9.2) supplemented with 15 mM TM-βCD. The electrophoretic runs were performed at 30°C under the application of 22 kV in a fused-silica capillary (50 µm id, 64.5 cm total length, 56 cm length to the detector). The method was validated for linearity of CyA (within 0.25–5.00 mg/mL) and CyH and IsoCyA (within 0.25–5.0%, w/w with respect to CyA), sensitivity (LOQ 5.0 µg/mL), accuracy, and precision. The applicability of the method was proved by analysis of a commercially available pharmaceutical (gelatin capsules).

Chiral separation of vinpocetine using cyclodextrin-modified micellar electrokinetic chromatography

A cyclodextrin-modified micellar electrokinetic chromatography (CD-MEKC) technique has been developed for enantioseparation of vinpocetine using an inexpensive 2-hydroxypropyl-β-CD (HP-β-CD) as the chiral selector (CS). The best chiral separation was achieved using 40 mM HP-β-CD as the CS in 50 mM phosphate buffer (pH 7.0) consisting of 40 mM sodium dodecyl sulfate (SDS) at a separation temperature and separation voltage of 25°C and 25 kV, respectively. To the author's best knowledge, this is the first CD-MEKC study able to successfully separate the four stereoisomer of vinpocetine in separation time of 9.5 min and resolution of 1.04-3.87.

Trace analysis of d-tyrosine in biological samples by microchip electrophoresis with laser induced fluorescence detection

A rapid and sensitive microchip electrophoresis (MCE) method with laser induced fluorescence (LIF) detection has been developed for the quantification of d-tyrosine (Tyr) in biological samples. The assay was performed using a MCE-LIF system with glass/poly(dimethylsiloxane) (PDMS) hybrid microchip after pre-column derivatization of amino acids with fluorescein isothiocyanate (FITC). Chiral separation of the derivatives was achieved by cyclodextrin-modified micellar electrokinetic chromatography (CD-MEKC) using γ-CD as chiral selector in the running buffer. d/ l-Tyr enantiomer was well separated in less than 140 s. The limit of detection ( S/ N = 3) was 3.3 × 10 −8 M. Using the present method, d-Tyr level in human plasma was found to vary significantly from normal humans to patients suffering from renal failure.