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@#In this paper, micellar electrokinetic chromatography (MEKC) with glucose-β-cyclodextrin (Glu-β-CD) as chiral selector and ionic liquid surfactant N-butyl-N-methyl pyrrolidine lauryl sulfate ([C4MP] [C12SO4]) micelles formed at low pH as a pseudo stationary phase was applied for the chiral separation of four acidic drugs naproxen,warfarin, ketoprofen and ibuprofen.Under the same conditions,significantly improved separation of tested drug enantiomers was achieved with the MEKC system based on [C4MP][C12SO4] compared with the system based on the conventional surfactant sodium dodecyl sulfate (SDS).Several primary parameters affecting enantioseparation such as type and proportion of organic modifier, concentration and pH of the running buffer, concentration of chiral selector,concentration of ionic liquid surfactant and applied voltage were systematically investigated.
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The present study determined five saponins in Xuesaitong Dropping Pills(XDP) by micellar electrokinetic chromatography(MEKC), and evaluated between-batch consistency by MEKC fingerprints and similarity analysis. A background buffer was composed of 20 mmol·L~(-1) sodium tetraborate-20 mmol·L~(-1) boric acid solution(pH 8.5), 55 mmol·L~(-1) sodium dodecyl sulfate(SDS), 23 mmol·L~(-1) β-cyclodextrin, and 13% isopropyl alcohol. All separations were performed at 25 ℃,20 kV and the detection wavelength was set at 203 nm. The separation channel was a fused silica capillary with a dimension of 75 μm I.D. and a total length of 50.2 cm(effective length of 40.0 cm). The contents of notoginsenoside R_1, and ginsenosides Rg_1, Re, Rb_1, Rd were determined with their quality control ranges set. The fingerprints of XDP were established and the between-batch consistency was evaluated by similarity analysis. The contents of five saponins from the 19 batches of XDP were stable in the fixed ranges. Statistical analysis was carried out on the results of multiple batches of samples, and the specific quality control ranges were recommended as follows: notoginsenoside R_1 21.92-34.16 mg·g~(-1), ginsenosides Rg_1 83.54-131.78 mg·g~(-1), ginsenosides Re 13.58-19.82 mg·g~(-1), ginsenosides Rb_1 89.40-129.90 mg·g~(-1), and ginsenosides Rd 22.34-35.67 mg·g~(-1). Eleven characteristic peaks were identified in the fingerprints. Five peaks, notoginsenoside R_1 and ginsenosides Rg_1, Re, Rb_1, Rd, were identified with reference standards. The similarities of the 19 batches of samples were all above 0.988, indicating good between-batch consistency. This method is green and simple, and can be used for the quantitative determination and quality evaluation of XDP. It can also provide references for the quality control of other Chinese medicinal dripping pills.
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Cromatografía Capilar Electrocinética Micelar , Medicamentos Herbarios Chinos , Micelas , Control de Calidad , SaponinasRESUMEN
OBJECTIVE: To establish a method for analysis of related substances in biapenem with micellar electrokinetic capillary chromatography(MEKC). METHODS: In order to improve the separation selectivity, a zwitterionic surfactant, 3-(N, N-dimethylhexadecylammonium)-propanesulfonate(PAPS) was used. The optimal separation conditions were as follows: the total length of the capillary was 48.5 cm (the effective length was 48 cm), the buffer was 90 mmol•L-1 tris(hydroxymethyl)aminomethane (tris)-phosphate buffer containing 17 mmol•L-1 PAPS and 3 mg•mL-1 polyoxyethylene 23 lauryl ether (Brij 35), the applied voltage was 22 kV, and the capillary temperature was controlled at 30℃. Further more, the specificity, linearity, precision, repeatability, stability and durability were studied. The contents of related substances in biapenem commercial samples were analyzed. RESULTS: The MEKC method, which was a comparable analysis method to HPLC, successfully separated the adjacent impurities of biapenem by using the zwitterionic surfactant PAPS. The specific test showed that this method was especially suitable for the detection of biapenem dimers A, B and open-ring compound. CONCLUSION: In this method, MEKC with zwitterionic surfactant is for the first time applied to the analysis of related substances in biapenem (amphoteric drugs). It provides a feasible analysis method with high sensitivity, good specificity and reproducibility for the quality control of biapenem.
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OBJECTIVE@#To establish a micellar electrokinetic capillary chromatography-based method for identification and quantitative detection of interleukin-12 (IL-12) and analysis of its unfolding process.@*METHODS@#An uncoated fused-silica capillary (inner diameter 50 μm) with a total length of 48.5 cm (40 cm to the detector) was used for the experiment. The factors influencing the separation efficiency of IL-12 were analyzed, and a standard curve of IL-12 concentration was established. The mixture of IL-12 and anti-IL-12 antibody was incubated in a water bath at 38 ℃ for 40 min, and capillary electrophoresis was then performed under the same conditions. The results were compared with those of IL-12 and anti-IL-12 antibody to identify IL-12. IL-12 and dithiothreitol (DTT) were incubated at 60 ℃ in water bath for different lengths of times, and the unfolding process of IL-12 was analyzed based on electrophoresis results of IL-12 in different states.@*RESULTS@#A micellar capillary electrophoresis on-line sweep method was established with 80 mmol/L borate (pH=9.3) containing 30 mmol/L sodium dodecyl sulfate (SDS) as the buffer solution. This system showed a good linear relationship between the peak area and the mass concentration of IL-12 with a linear correlation coefficient of 0.9991 within the linear range of 2 to 120 ng/L. As the incubation time of IL-12 and DTT prolonged, the disulfide bond of IL-12 gradually opened and resulted in distinct changes in the protein peak.@*CONCLUSIONS@#This capillary electrophoresis-based method is simple and sensitive for IL-2 analysis and allows rapid detection of changes in IL-12 content in the setting of tumors and analysis of the possible causes.
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Estudos envolvendo os glicocorticoides merecem destaque devido a serem hormônios responsáveis pela transferência de informações e instruções às células, desta forma regulando o metabolismo, desenvolvimento, crescimento, função imune e também auxiliam no controle das funções tanto reprodutivas quanto tecidual. Estes também são sintetizados e amplamente utilizados com finalidade terapêutica processos alérgicos, tratamento de doenças autoimunes, em transplantes no pré-operatório e/ou pós-operatório-, devido a sua eficiente ação como imunossupressores e anti-inflamatórios. Os dois primeiros capítulos deste trabalho exibem uma revisão da literatura com foco em considerações gerais sobre os glicocorticoides, metodologias empregadas na análise destes hormônios e fundamentos da eletroforese capilar. Na sequência, o quarto capitulo, mostra a otimização da separação de 17 glicocorticoides utilizando cromatografia eletrocinética micelar devido a alto grau hidrofóbico dos analitos. Para tal, a composição do eletrólito consistiu em 20mM de tetraborato de sódio (pH=9.3) e 30 mM de dodecil sulfato de sódio (como surfactante), e a interação soluto-micela e, portanto, retenção do soluto, foi manipulada com a adição (volume/volume) de solventes orgânicos na composição de até 20% acetonitrila (ACN), 20% etanol (EtOH) e 1% tetrahidrofurano (THF), a qual se baseia num modelo de desenho de misturas (totalizando dez diferentes eletrólitos), e através desta abordagem um ótimo de separação foi obtido (13,3% EtOH, 3,3% ACN e 0,17% THF). A melhor condição de separação foi testada qualitativamente numa amostra de urina de um voluntário que faz uso contínuo de prednisona como terapia corticoidal. As misturas de solventes estudadas neste trabalho afetam a solubilidade dos hormônios na fase aquosa e a estrutura micelar também sofre grande impacto,principalmente na camada de solvatação. O quarto capítulo busca racionalizar tais efeitos através da obtenção de descritores, e as informações contidas nos descritores hidrofóbicos e hidrofílicos são sempre relevantes e contribuem nas correlações encontradas. Obteve três grupos de comportamento distinto, onde a capacidade doadora e aceptora de prótons para a realização de ligações de hidrogênios foram as interações consideradas as mais relevantes para o comportamento observado da separação. E o capítulo final, apresenta possibilidades de aproveitamento no controle de qualidade na indústria farmacêutica, métodos baseados na injeção e tensão inversas foram propostos a fim de ganho de tempo de análise (máximo de 5 minutos), estes foram validados seguindo o protocolo preconizado pela ANVISA (Agência Nacional de Vigilância Sanitária) nos parâmetros: precisão, exatidão, seletividade, linearidade, limites de detecção e quantificação e robustez; e aplicados na quantificação de quatro (diferentes formulações comerciais contendo glicocorticoides (prednisona 20 mg, betametasona 4 mg, furoato de mometasona 200 mcg e dipropionato de beclometasona 200 mcg)
Studies involving glucocorticoids deserve to be highlighted because they are hormones responsible for the transfer of information and instructions to cells, thus regulating metabolism, development, growth, immune function and also assist in the control of both reproductive and tissue functions. These are also synthesized and widely used for therapeutic purposes - allergic processes, treatment of autoimmune diseases, in preoperative and/or postoperative transplants - due to their efficient action as immunosuppressants and anti-inflammatories. The first two chapters of this paper present a review of the literature focusing on general considerations about glucocorticoids, methodologies used in the analysis of these hormones and fundamentals of capillary electrophoresis. Subsequently, the fourth chapter shows the optimization of the separation of 17 glucocorticoids using micellar electrokinetic chromatography due to the high hydrophobic degree of the analytes. To this end, the electrolyte composition consisted of 20 mM sodium tetraborate (pH = 9.3) and 30 mM sodium dodecyl sulfate (as a surfactant), and the solute-micelle interaction and therefore solute retention was manipulated with organic solvent in the composition of up to 20% acetonitrile (ACN), 20% ethanol (EtOH) and 1% tetrahydrofuran (THF), which is based on a mixture design model (totaling ten different electrolytes), and through this approach an optimal separation was obtained (13.3% EtOH, 3.3% ACN and 0.17% THF). The best separation condition was qualitatively tested in a urine sample from a volunteer who makes continuous use of prednisone as corticosteroid therapy. The solvent mixtures studied in this work affect the solubility of the hormones in the aqueous phase and the micellar structure also has a great impact, especially on the solvation layer. The fourth chapter seeks to rationalize these effects by obtainingdescriptors, and the information contained in the hydrophobic and hydrophilic descriptors is always relevant and contributes to the correlations found. It obtained three groups of distinct behavior, where the donor and acceptor capacity of protons for the realization of hydrogen bonds were the interactions considered the most relevant for the observed behavior of the separation. And the final chapter presents possibilities of use in quality control in the pharmaceutical industry, methods based on injection and reverse voltage were proposed in order to gain analysis time (maximum of 5 minutes), these were validated following the protocol recommended by ANVISA (Brazilian National Agency of Sanitary Surveillance) in the parameters: precision, accuracy, selectivity, linearity, limits of detection and quantification and robustness; and applied in the quantification of four different commercial formulations containing glucocorticoids (prednisone 20 mg, betamethasone 4 mg, mometasone furoate 200 mcg and beclomethasone dipropionate 200 mcg)
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Electroforesis Capilar , Composición de Medicamentos , Glucocorticoides/análisis , Esteroides , Cromatografía Capilar Electrocinética Micelar/métodosRESUMEN
Objective To establish a micellar electrokinetic capillary chromatography method for content determination of geniposide in Biyuanshu Oral Liquid. Methods Acetaminophen was used as an internal standard, and the separation was performed on an uncoated fused silica capillary of 52 cm × 50 μm ID (42 cm effective length) with the separation voltage of 25.0 kV. The running buffer contained 50 mmol/L borax, 100 mmol/L sodium dodecyl sulfate and 15％ acetonitrile (pH=10). The sample was injected by pressure (10 s, 0.5 psi) and detected at 238 nm. Results Geniposide was in good linearity range of 15.02–320.48 μg/mL (r=0.9995). The repeatability (low, medium and high concentration of samples) and intermediate precision assays gave satisfactory RSD values of less than 1.77％and 2.01％, respectively. The average recovery of geniposide in Biyuanshu Oral Liquid was 97.50％ and the RSD was 4.43％. The contents of geniposide determined by micellar electrokinetic capillary chromatography were in accordance with the results of HPLC analysis. Conclusion The method is simple, fast, accurate and precise, which can be used for the content determination of geniposide in Biyuanshu Oral Liquid.
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Abstract A rapid and sensitive micellar electrokinetic capillary chromatography method with UV photodiode-array detection was developed for the simultaneous determination of atorvastatin and ezetimibe in fixed dose drug combination. Experimental conditions such as buffer concentration and pH, surfactant concentration, system temperature, applied voltage, injection parameters were optimized in order to improve the efficiency of the separation. The best results were obtained when using fused silica capillary (48 cm length X 50 µm ID) and 25 mM borate buffer electrolyte at pH 9.3 containing 25 mM SDS, + 30 kV applied voltage, 20 ºC system temperature. The separation was achieved in approximately 2 minutes, with a resolution of 7.02, the order of migration being atorvastatin followed by ezetimibe. The analytical performance of the method was verified with regard to linearity, precision, robustness and the limit of detection and quantification were calculated.
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Cromatografía Capilar Electrocinética Micelar/métodos , Ezetimiba/administración & dosificación , Atorvastatina/administración & dosificación , Preparaciones Farmacéuticas/análisis , Fraccionamiento de la Dosis de RadiaciónRESUMEN
Bioanalytical methods are widely used for quantitative estimation of drugs and their metabolites in physiological matrices. These methods could be applied to studies in areas of human clinical pharma-cology and toxicology. The major bioanalytical services are method development, method validation and sample analysis (method application). Various methods such as GC, LC-MS/MS, HPLC, HPTLC, micellar electrokinetic chromatography, and UFLC have been used in laboratories for the qualitative and quan-titative analysis of carbamazepine in biological samples throughout all phases of clinical research and quality control. The article incorporates various reported methods developed to help analysts in choosing crucial parameters for new method development of carbamazepine and its derivatives and also enu-merates metabolites, and impurities reported so far.
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Objective: To establish a rapid method for detecting acetylbritannilactone (ABL) by online sweeping-micellar electrokinetic chromatography (MEKC) and to elevate the sensitivity of the detection. Methods: The combination of online sweeping technique with MEKC was used to determine the content of ABL in the extract of Inula britannica in plasma of rats. Results: ABL was completely separated within 15 min in running buffer and sample buffer. The optimal conditions were as follows: on uncoated fused quartz silica capillary, with separation voltage of 23 kV, capillary temperature of 25 °C, and detection wavelength of 195 nm. The regression equations revealed good linear relationships between the peak area and concentration of ABL (r = 0.998), with the detection limits of 0.005-0.15 mg/mL. The relative standard deviations of migration time and peak areas for intra- and inter-batch were < 2.45% and < 2.26%, respectively. The recovery rate of this method was 96.3%-97.2%. Conclusion: This method provides some advantages in separation speed, testing sensitivity, and operating convenience, with low sample and reagent consumption. The online sweeping-MEKC is an effective method for pharmacokinetic study and analysis on tracing biological samples. © 2014 Tianjin Press of Chinese Herbal Medicines.
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OBJECTIVE:To develop a simple micellar electrokinetic capillary chromatography for determination of theophylline concentration in human plasma. METHODS: The determination of plasma sample(without any pretreatment) was performed on un-coated fused-silica capillaries (at an effective length of 21 cm) in which the cathodic buffer solution (solution A) was 15 mmol?L-1 borax(pH=10) and the running buffer solution(solution B) was sodium dodecyl sufate (SDS)(200 mmol?L-1) -contained solution A. The sample was injected into capillary by pressure with separation voltage of 12 kV;the UV detection wavelength was set at 280 nm;quantitation was performed by external standard peak area method. RESULTS: The average determination time of each sample was 11 min;the linear range of theophylline was 1.25～40 ?g?mL-1 with its lowest detectable limit at 0.5 ?g?mL-1;the average methodological recovery was 90.33%(RSD=2.51%);the intra-day RSD was 2.34%～3.36% and the inter-day RSD was 2.03%～6.51%,respectively. CONCLUSION: The developed method is simple,rapid and sensitive and it is applicable for the therapeutic drug monitoring of theophylline.
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OBJECTIVE: A micellar electrokinetic capillary chromatography(MECC) method was established for determination of benzoic acid and salicylic acid in Zuguang san.METHODS: An uncoated silica capillary(75 ?m?55 cm,effective length 47 cm) was used with 20 mmol?L-1 borate-30 mmol?L-1 SDS-4% methanol(pH 9.8) as the running buffer solution.The internal standard was antiscorbic acid.The detection wavelength was set at 250 nm.RESULTS:The calibration curves were linear in the range of 120～320 ?g?mL-1 for benzoic acid(r=0.999 8) and 80～320 ?g?mL-1(r=0.999 5) for salicylic acid.The average recoveries were 98.37% and 100.69%,respectively and the RSD were 4.47%(n=9) and 1.61%(n=9),respectively.CONCLUSION: MECC method was proved to be simple,sensitive and reproducible with high recovery rate and it is applicable for the determination of the contents of benzoic acid and salicylic acid in Zuguang san.
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Resumen. El envenenamiento por escorpiones es un problema de salud pública en la zona suroccidental de Venezuela. El Tityus zulianus es uno de los escorpiones que causa, especialmente en niños, edema pulmonar e insuficiencia cardíaca que pueden ser fatales y que se han atribuido en parte a una gran descarga simpática. La administración intraperitoneal de (20 µg/g peso) de veneno del T. zulianus a ratones anestesiados durante microdiálisis subcutánea provocó aumento de secreciones, dificultad respiratoria, convulsiones y muerte entre 30 min a 2 h. En los dializados recolectados antes y después de recibir el veneno, se analizaron 7 aminoácidos por electroforesis capilar con detección mediante fluorescencia inducida por láser (EC-DFIL). Se encontró un aumento en arginina (39%), fenilalanina (40%) y glutamato (94%), sin cambios en valina, serina y aspartato, en los animales que recibieron veneno con respecto a sus valores pretratamiento. Estos cambios porcentuales fueron significativos al comparar veneno vs. vehículo después de la inyección; y antes vs. después de recibir el veneno. Para este momento no hay una explicación clara del significado de estos aumentos de aminoácidos específicos. Se requieren nuevos estudios para conocer si estos cambios bioquímicos tienen o no una relación con los mecanismos de acción molecular del veneno o algunos de sus componentes y/o con las manifestaciones clínicas. De acuerdo con la literatura revisada, este es el primer reporte de la combinación de microdiálisis subcutánea y EC-DFIL en el estudio in vivo del emponzoñamiento por escorpiones en ratones.
Abstract. Scorpion human envenoming is a public health hazard in the southwest of Venezuela. Tityus zulianus is one of the scorpion species whose venom causes lung edema and cardiac failure in children. These occasionally deadly manifestations have been attributed to a massive sympathetic discharge. The intraperitoneal administration of T. zulianus venom (20 µg/g mouse) to anesthetized mice during subcutaneous microdialysis caused increased secretions, dyspnea, seizures and death between 30 min to 2 h. Seven amino acids were analyzed by capillary electrophoresis with laser induced fluorescence detection (CE-LIFD) in the collected samples before and after the venom administration. We found an increase of arginine (39%), phenylalanine (40%) and glutamate (94%), with no changes in valine, serine and aspartate, changes were significant when the injection of venom and vehicle were compared and before vs after venom injection. Further investigation is needed to know if the observed changes could be related to the molecular mechanisms of the venom or some of its components and therefore with the envenoming symptoms. To our knowledge, this is the first report with subcutaneous microdialysis and CE-LIFD coupling in scorpion envenomation studies in vivo, in mice.
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Animales , Masculino , Ratones , Aminoácidos/biosíntesis , Venenos de Escorpión/administración & dosificación , Aminoácidos/análisis , Electroforesis Capilar , Inyecciones Intraperitoneales , Microdiálisis/métodos , PielRESUMEN
This article reviewed the feature and the advances in the application of micellar electrokinetic capillary chromatography(MECC),an important mean of high performance capillary electrophoresis(HPCE),for the analysis of complicated component of medicine,.The analysis of multi-component of routine medicine,chiral drugs,illicit drugs,Chinese medicine,drug and its metabolites were introduced.
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Objective: To establish a method for the determination of quercetin and kaempferol in Ginkgo biloba extract. Methods: A simple, rapid and reproducible supercritical fluid extract (SFE) and capillary electrophoretic method were developed for it. SFE was selected by orthogonal design method. SFE conditions: temperature 60 ℃, pressure 42 mPa, static extraction 4 min, dynamic extraction volume 4 ml and 0.2 ml ethanol as modifier. MECC conditions were 55 cm?75 ?m fused silica capillary column. The running buffer contained 25 mmol/L sodium dihydrophosphate, 6.25 mmol/L sodium borate (pH= 8.5) and 35 mmol/L sodium dodecyl sulfate(SDS). The detection wave length was at 254 nm. Results: The influence of modifier on extraction rate in SFE was the most. The linear range for quercetin and kaempferol was 21.2 106.0, 20.0 100.0 ?g/ml, respectively, when cinnamic acid was in an internal standard. The recovery of quercetin for 40, 60 and 80 ?l was 93.87%, 94.02% and 94.10%, respectively. The recovery of kaempferol for 40, 60 and 80 ?l was 94.50%, 94.17% and 94.25%, respectively. Conclusion: SFE MECC is convenient and accurate method, and can be used as a measure of quality control for Ginkgo biloba extract. [
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Object To isolate and identify the four active constituents, chrysophanol, emodin, rhaponticin and rhein in rhubarb Methods Micellar electrokinetic capillary chromatography (MECC) with a buffer solution of 5 mmol/L H 3BO 3 NaOH containing 20 mmol/L of sodium deoxycholate (SDC) (pH=10) was used for their separation at a voltage of 17 kv Results The four active constituents were completely separated within 5 min Conclusion This method was found to be simple and rapid and gave satisfactory results
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Objective To develop a capillary electrophoresis method for the determination of methamidopho residue in water.Methods Methamidopho in water was determined by micellar electrokinetic capillary chromatograph.The qualitative analysis was done by retention time and quantitative analysis by external standard curve.Results The optimum conditions for methamidopho determination were: pH= 7.5,20 mmol/L phosphate buffer,30 mmol/L SDS,wavelength of 254 nm,separate voltage 25 kV,inject duration of 9 s.The linear range was 1.0-200 ?g/ml,the correlation coefficient was 0.999 1,the rates of recovery were 91.5%-105.2%,the relative standard deviations were 1.35%-3.24% for this method.The limit of determination was 0.007 1 ?g/ml(capillary volume was 3 ?l).Conclusion This method was sensitive,accurate and applicable to rapid the determination of methamidopho.
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OBJECTIVE:To determine serum concentrations of phenbarbital(PB),phenytoin sodium(PHT),carbamazepine(CBZ)by micellar electrokinetic capillary chromatograph(MEKC).METHODS:The determination was performed on elastic quartz column(37 cm?50 ?m)with barbital as internal standard.The buffer solution consisted of 10 mmol?L-1 phosphate sodium buffer-30 mmol?L-1 SDS sodium(micellar phase)-15% methanol at a constant voltage of 18 kV.The detection wavelength was set at 210 nm.RESULTS:The linear concentration ranges of PB and PHT were 2.5～80 ?g?mL-1,with detective concentration of 0.1 ?g?mL-1;the linear range and detective concentration of CBZ were 1～32 ?g?mL-1 and 0.2 ?g?mL-1,respectively.The average recoveries of the PB,PHT,and CBZ were 99.90%,99.91%,and 99.42%,respectively.CONCLUSION:The method is rapid,simple,accurate,sensitive and reproducible,and suitable for clinical monitoring of drug level and pharmaceutical study.
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Objective To develop a method for simultaneous determination of benzodiazepines in human whole blood by SPE-Micellar electrokinetic capillary chromatography.Methods With the Clenbuterol as internal standard,Oasis column was used to extract the drugs from whole blood.The separation was performed on a fused-silica capillary of 75?m ID?50.2cm(40cm of effect length).The running buffers were sequentially used as 15mmol/L phosphates→15mmol/L sodium borate(pH8.2)→30 mmol/L SDS,and 18% methanol served as an organic modifier.Sample solution was injected with pressure mode,and the running voltage was 25kV.The detection wavelength was set at 230nm.Results The linear ranges of the calibration curves were from 0.02 to 1.6?g/ml,and the limits of detection ranged between 5 ng/ml and 50 ng/ml.The within-day and between-day precision was less than 12%.Conclusion The method developed for determination of benzodiazepines in human whole blood is effective,simple and reliable,with which 9 benzodiazepines may be simultaneously separated.