Simultaneous chiral separation of ephedrine alkaloids by MEKC-ESI-MS using polymeric surfactant II: application in dietary supplements

Chiral MEKC-MS method was utilized for separation, identification, and quantitation of ten enantiomers of ephedrine and related compounds. Enantioselective separations of all ephedrine alkaloids were accomplished through a combination of polysodium N-undecenoxycarbonyl-L-leucinate (poly-L-SUCL) with 30% v/v ACN. Interestingly, the more hydrophilic stereoisomers were eluted later than the hydrophobic ones indicating that hydrogen bonding interactions are much stronger than hydrophobic interactions in the presence of ACN in chiral MEKC. The method was validated in terms of linearity, LOD, LOQ, precision and robustness. The method was finally used in the analysis of three standard reference materials (SRMs). Results of (-)-ephedrine ranged from 12.49 to 0.24 mg/g, for (+)-pseudoephedrine from 4.04 to 0.019 mg/g, for (-)-norephedrine from 0.36 to 0.0031 mg/g, for (+)-norpseudoephedrine from 0.68 to 0.0052 mg/g, for (-)-methylephedrine from 1.18 to 0.0092 mg/g and for (+)-methylpseudoephedrine from 0.086 to 0.00037 mg/g in the SRMs.     

Chiral separation of polychlorinated biphenyls using a combination of hydroxypropyl-gamma-cyclodextrin and a polymeric chiral surfactant

Chiral separation of moderately to highly hydrophobic polychlorinated biphenyls (PCBs) using a conventional chiral micelle or a polymeric chiral surfactant, as the single chiral selector is very difficult since the hydrophobic interactions between the chiral PCB and the monomeric or polymeric surfactant is very strong. Combined use of a polymeric chiral surfactant, polysodium N-undecanoyl-D-valinate (poly-D-SUV) with hydroxypropyl-gamma-cyclodextrin (HP-gamma-CD) was successful in cyclodextrin modified electrokinetic chromatography (CD-EKC) enantioseparation of PCB congeners. Addition of HP-gamma-CD to the background electrolyte containing poly-D-SUV functioned to improved chiral resolution for the PCBs and reduce the analysis time for these congeners. In addition, concentration of methanol, concentration of 2-(N-cyclohexylamino) ethanesulfonic acid (CHES) buffer and separation voltage was also varied to optimize multicomponent separation of five chiral PCBs. Simultaneous separation and enantioseparation of all five PCBs was possible in less than 50 min under optimized conditions that requires a 5 mM CHES solution buffered at about pH 10 with 1.5% w/v (ca. 60 mM) poly-D-SUV and 16 mM HP-gamma-CD. In addition, 1 M urea and 20% v/v methanol should be added as organic modifier and the capillary temperature maintained at 45 degrees C. As expected the polymeric surfactant showed improved chiral resolution of PCBs over conventional micelles of SUV. Under optimized conditions, when CD-EKC of chiral PCBs using poly-D-SUV was compared to sodium dodecyl sulfate (SDS), better resolution, higher efficiency and shorter analysis time was achieved with poly-D-SUV.     

Capillary electrophoretic chiral separation of Cinchona alkaloids using a cyclodextrin selector

A new capillary electrophoretic method for the chiral separation of four major Cinchona alkaloids (quinine/quinidine and cinchonine/cinchonidine) was developed using heptakis-(2,6-di-O-methyl)-beta-cyclodextrin as the chiral selector. The inner walls of the separation capillary were modified with a thin polyacrylamide layer, which substantially reduced the electroosmotic flow and improved the chiral resolution and the reproducibility of the migration time of the analytes. Various operation parameters were optimised, including the pH, the capillary temperature, the concentration of the background electrolyte, and the concentration of the chiral selector. Baseline separation of the two diastereomer pairs was achieved in 12 minutes in ammonium acetate background electrolyte pH 5.0 with addition of cyclodextrin in a concentration of 3 mM or higher.     

用聚合表面活性剂作增敏试剂分光光度测定钼和钨

采用表面活性剂的分光光度分析法应用日趋广泛。这方面的研究工作虽极活跃,但较多工作限于已有经典表面活性剂的研究与应用,为分析化学的特定需要设计与合成新型表面活性剂的研究尚少。作者曾用环氧氯丙烷和长链烷基叔胺合成了以聚乙二醇为主链含有若干个带长链烷基的季氮支链的低聚合表面活性剂——聚(氧化丙烯)-α-十八烷基二甲基氯化铵(PPOSA)。实验表明,这类表面活性剂兼有阳离子和非离子表面活性剂的某些特性。本文     

Simultaneous determination of rare earth elements by using substoichiometric separation of lanthanium

An analytical method for the simultaneous determination of rare earth elements by using substoichiometry with two chelating agents, a complexon and an extracting agent, is proposed. The principle of the method is that the element which has the lowest stability constant with the complexon is separated substoichiometrically, while other elements which have higher stability constants are separated quantitatively by a single extraction. This method was applied to neutron activation analysis of rare earth elements. Lanthanum, europium and terbium in orchard leaves were determined simultaneously by using substoichiometric separation with DTPA and TTA.     

Determination of ephedrine alkaloids by capillary electrophoresis

A simple and rapid method for the simultaneous determination of six ephedrine alkaloids (ephedrine, pseudoephedrine, norephedrine, norpseudoephedrine, methylephedrine and methylpseudoephedrine) in Ephedrae herba by capillary electrophoresis was developed. A buffer solution that contained 0.005 M barium hydroxide and 0.02 M isoleucine and adjusted to pH 10.0 with ammonia solution was found to be the most suitable electrolyte for this separation. The contents of the six alkaloids in the crude drug of Ephedrae herba could be easily determined.     

Simultaneous separation and determination of hydrocarbons and organochlorine compounds by using a two-step microcolumn.

The simultaneous separation and determination of a mixture of hydrocarbons and organochlorine compounds was successfully carried out by using sorption chromatography on a two-step microcolumn of silica and aluminium oxide for their fractionation, and a dual detector system. In addition to the separation and identification of hydrocarbons and heterocompounds containing nitrogen, oxygen and sulphur atoms, separation and identification of chlorinated hydrocarbons (dichlorobenzenes, p-chlorotoluene, hexachlorobutadiene, 1,2,4-trichlorobenzene and 2-chloronaphthalene), pesticides (chlorpicrin, aldrin, lindane, alpha- and beta-benzene hexachloride (BHC), endrin, dieldrin, endosulphan, methoxychlor) and herbicides (propanil, dichlobenil, trifluralin, difolatan) were achieved in mixtures containing polychlorinated biphenyl, strobane and chlordane.     

Simultaneous chiral separation of triadimefon and triadimenol by sulfated beta-cyclodextrin-mediated capillary electrophoresis

Enantiomeric separation of two triazole fungicides, triadimefon and triadimenol, was investigated in sulfated beta-cyclodextrin (sulfated beta-CD)-mediated capillary electrophoresis (CE) systems. It was found that, at pH 2-4, sulfated beta-CD exhibited strong chiral recognition towards both triadimefon and triadimenol. The enantiorecognition was believed to result from the multiple interactions between sulfated beta-CD and the analytes, which included inclusion effect, electrostatic interaction, and hydrogen bonding. Under optimal conditions (phosphate buffer with 2% sulfated beta-CD, pH 2.5), simultaneous resolution of all chiral isomers of triadimefon and triadimenol was achieved in less than half an hour. In conjunction with solvent extraction and subsequent enrichment by solid-phase extraction (SPE), this new enantioseparation method was applied successfully in the study of stereoselectivity associated with the biotransformation of triadimefon to triadimenol by soil microorganisms. The present methodology was superior to the commonly adopted chiral gas chromatography (GC) approach in that a very mild procedure was involved from sample extraction to the ultimate chiral separation. Thus, the disturbance of the enantiomeric distribution patterns of the original soil samples by heat stress was an unlikely scenario. Furthermore, it was discovered that, owing to the unique selectivity of the present separation strategy, there was virtually no interference from the soil matrix, which led to improvements in both sensitivity and selectivity in real sample determination.     

Separation and determination of ephedrine alkaloids and tetramethylpyrazine in Ephedra sinica Stapf by gas chromatography-mass spectrometry

A simple, sensitive, and reliable method using gas chromatography (GC)-mass spectrometry (MS) is developed for the simultaneous determination of ephedrine alkaloids and 2,3,5,6-tetramethylpyrazine (TMP) in Ephedra sinica Stapf. The sample is extracted with ethyl ether and submitted to GC-MS for identification and quantitation without derivatization. The column used for GC is an HP-5 (30.0 m x 250 microm x 0.25 microm, 5% phenyl methyl siloxane), and the carrier gas is helium. The detection limits for ephedrine, pseudoephedrine, and TMP are 0.4 ng 0.7 ng, and 0.02 ng (signal-to-noise ratio of 3), respectively. The reproducibility of the total procedure is proved to be acceptable (RSD < 2%), and the recoveries are above 93%.     

Bupropion hydrochloride: the development of a chiral separation using an ovomucoid column

The separation of bupropion enantiomers on an ovomucoid stationary phase was investigated. The mobile- and stationary-phase parameters that may influence the separation were identified. The parameters that were studied include: type and concentration of organic modifier, mobile phase pH, ionic strength, type of buffer, and column temperature, as well as the effect that the amount of sample injected had on the separation. The optimized chiral separation baseline-resolved the enantiomers in less than 10 min. Calibration curves for a standard were linear over a range of 0.27-53.0 microg/g (ppm) with a correlation coefficient of 0.999 for both enantiomers. A detection limit of 0.13 microg/g and a quantitation limit of 0.27 microg/g were also found. The system precision of the method was 0.2%.     

Rapid chiral separation and impurity determination of levofloxacin by ligand-exchange chromatography

A sensitive, simple, and accurate method for determination of levofloxacin and its (R)-enantiomer was developed to determine the chiral impurity of levofloxacin in Cravit Tablets material by ligand-exchange high performance liquid chromatography. The effects of different kinds of ligands, concentration of ligands in mobile phase, organic modifier, pH of mobile phase, and temperature on enantioseparation were investigated and evaluated. Chiral separation was performed on a conventional C₁₈ column, where the mobile phase consisted of a methanol-water solution (containing10 mmol L⁻¹l-leucine and 5 mmol L⁻¹ copper sulfate) (88:12, v/v) and its flow-rate was set at 1.0 mL min⁻¹. The conventional C₁₈ column offers baseline separation of two enantiomers with a resolution of 2.4 in less than 20 min. Thermodynamic data (ΔΔH and ΔΔS) obtained by Van’t Hoff plots revealed the chiral separation is an enthalpy-controlled process. The standard curves showed excellent linearity over the concentration range from 0.5 to 400 mg L⁻¹ for levofloxacin and its (R)-enantiomer. The linear correlation equations are: y = 1.33 × 10⁵x + 6297 (r = 0.9991) and y = 1.34 × 10⁵x + 3565 (r = 0.9997), respectively. The relative standard deviation (RSD) of the method was below 2.3% (n = 3).     

Enantioseparations by capillary electrophoresis using chiral glycosidic surfactants. I. Evaluation of cyclohexyl-pentyl-beta-D-maltoside surfactant.

Chiral cyclohexyl-pentyl-beta-D-maltoside (CYMAL-5) surfactant was evaluated in the enantioseparation of charged racemic species by capillary electrophoresis. CYMAL-5 is a glycosidic surfactant (GS) with a chiral maltose polar head group and a cyclohexyl-pentyl hydrophobic tail. At concentrations above its critical micellar concentration (CMC), CYMAL-5 produces neutral micelles in aqueous media. The neutral micelles migrate at the velocity of the electroosmotic flow (EOF). As expected, the CYMAL-5 system was only useful for the enantioseparation of charged chiral solutes. The enantioresolution of the CYMAL-5 can be manipulated over a wide range of electrolyte composition, e.g., pH, ionic strength and surfactant concentration. In the presence of EOF, and in all cases, there is an optimum surfactant concentration for maximum enantioresolution, which is located at low surfactant concentration for strongly hydrophobic solutes and at high surfactant concentration for relatively hydrophilic solutes. The presence of an optimum surfactant concentration for maximum enantioresolution is attributed to the EOF. At low pH values where the EOF is negligible, enantioresolution increased with increasing surfactant concentration in the useful concentration range in a way similar to chromatography.     

Simultaneous chiral analyses of multiple analytes: case studies, implications and method development considerations

The field of chiral separations had a modest beginning some two decades ago. However, due to rapid technological advancement coupled with simultaneous availability of innovative chiral stationary phases and novel chiral derivatization agents, the field of chiral separations has now totally outpaced many other separation fields. Keeping pace with rapid changes in the field of chiral separations, investigators continue to add stereoselective pharmacokinetic, pharmacodynamic, pharmacologic and toxicological data of new and/or marketed racemic compounds to the literature. Examination of the evolution of chiral separations suggests that in the beginning many investigators attempted to separate and quantify a single pair of enantiomers, adopting either direct (separation made on a chiral stationary phase) or indirect (separation made following precolumn conversion of enantiomers to corresponding diastereomers) approaches. However, more recent trends in chiral separations suggest that investigators are attempting to separate and quantify multiple pairs of enantiomers with available technologies. Added to this, some interesting trends have been observed in many of the recently reported chiral applications, including preferences regarding internal standard selection, mobile phase contents and composition, sorting out issues with mass spectrometric detection, determination of elution order, analytical manipulations of metabolite(s) without reference standards and addressing some specificity-related issues. This review mainly focuses on chiral separations involving multiple chiral analytes and attempts to justify the need for such chiral separations involving multiple analytes. In this context, several cases studies are described on the utility and applicability of such chiral separations under discrete headings to provide an account to the readership on the implications of such tasks. The topics of case studies covered in this review include: (a) therapy markers--differentiation from drug abuse and/or applicability in forensics; (b) role in pharmacogenetic/polymorphic evaluation; (c) monitoring and understanding the role of parent and active metabolite(s) in clinical and preclinical investigations; (d) exploration on the pharmacokinetic utility of an active chiral metabolite vis-a-vis the racemic parent moiety; (e) understanding the chirality play in delineating peculiar toxic effects; (f) exploration of chiral inversion phenomenon, and understanding the role of stereoselective metabolism. For the further benefit of readership, some select examples (n = 19) of the separation of multiple chiral analytes with appropriate information on chromatography, detection system, validation parameters and applicable conclusion are also provided. Finally, the review covers some useful considerations for method development involving multiple chiral analytes.     

Rapid method development for chiral separation in drug discovery using sample pooling and supercritical fluid chromatography-mass spectrometry

A novel strategy for rapid chiral method development has been implemented using sample pooling and supercritical fluid chromatography-mass spectrometry (SFC-MS) on four chiral stationary phases, namely Chiralpak AD and AS, and Chiralcel OJ and OD, and eight different modifier concentrations (5 to 40% methanol-0.2% isopropylamine). The screening is performed under an outlet pressure of 110 bar at 35 degrees C, and at a flow-rate of 2.5 ml/min for the initial 20 min and then ramped up to 4 ml/min and held for 4.5 min to elute all solutes from the column. The entire process is fully automated from injection to data processing, and operates unattended for 15 h overnight to obtain optimal chiral separation for multiple compounds. A unique feature of using SFC-MS to monitor chiral synthesis is the negligible interferences from achiral impurities. In addition, with SFC-MS, enantiomeric excess can be determined with much lower detection limits than UV and much shorter analysis times compared to normal-phase/reversed-phase liquid chromatography.     

Rapid method development for chiral separation in drug discovery using multi-column parallel screening and circular dichroism signal pooling

A novel strategy for rapid chiral method development has been developed using multi-column parallel screening and circular dichroism (CD) signal pooling. Described is the first use of a customized HPLC system that integrates an HPLC auto-sampler, one pump and five divided channels with five columns and five UV detectors to screen five chiral stationary phases (CSPs) simultaneously in parallel. A high-pressure semi-prep on-line pre-filter, a six-port manifold and five individually adjusted backpressure restrictors were installed in the system which allowed the sample and mobile phase to be evenly distributed over the five columns and UV detectors. The five CSPs, namely Chiralpak AD and AS, Chiralcel OJ and OD and Whelk-O1, were screened. The system guarantees a five-fold increase in speed for chiral column scouting compared with the widely used automated sequential column switching approach, and does not have the limitations of the coupled column screening approach for enantiomers whose elution order could be reversed on CSPs. Furthermore, the five channels after the UV detectors were recombined using a reversed flow splitter into a CD detector. The pooled CD signal from the five channels was recorded to track the elution order of the resolved enantiomers and to determine their sign, positive or negative. The signal pooling allows for the effective use of a single CD detector for multiple columns since unresolved racemate has little CD signal, and observing the sign of CD signal for one of the two enantiomer UV peaks is sufficient for tracking the enantiomeric elution order.     

Simultaneous determination of metal traces by ICP-MS in environmental waters using SPE preconcentration on different polymeric sorbents

This paper deals with multielement profiling of microlements in the form of their isotopes ⁹Be, ⁵¹V, ⁵⁹Co, ⁶⁰Ni, ⁸⁹Y, ¹¹¹Cd, ²⁰⁸Pb, ²³²Th, and ²³⁸U. After their complexation by 4-(2-pyridylazo)resorcinol (PAR), 3,4-dihydroxy-9,10-dioxo-2-anthracenesulfonic acid sodium salt (ALS), 8-hydroxyquinoline-5-sulphonic acid (8-HQS), and ammonium pyrrolidinedithiocarbamate (APDC), the elements were preconcentrated and separated on Amberlite XAD-16 and Amberlite SDB-L prior to their analysis by inductively coupled plasma mass spectrometry. Various parameters such as pH, eluent type and volume, presence of surfactants and volume, and matrix effects on the retention of analytes were examined. Relative standard deviation and recovery values for four replicate determinations under optimal condition were in the range of 0.2?C3.6 % and 59?C98 %. The proposed method was applied to the determination of elements in lake water sample and industrial water. Recovery experiments with spiked water samples were performed.     

Chiral separation of amino acids using a chiral crown ether by impregnation on a polymeric support and monoamine modified silica gel

A chiral monoaza-15-crown-5 ether derivative was prepared from l-Leucinol and used as a chiral stationary phase. The new chiral stationary phases CSP-1 and CSP-2 were employed in separating the enantiomers of the sodium and potassium salts of amino acids. The sodium and potassium salt of the d-enantiomers of all amino acids (PhyAlaNa, PhyAlaK and PhyGlyNa, PhyGlyK, and TrpNa, TrpK) show higher selectivity than the l-enantiomers for both CSP-1 and CSP-2.