RESUMEN
Omeprazole (OME) is a proton pump inhibitor used to treat gastroesophageal reflux disease associated conditions. The current study presents an Analytical Quality by Design-based approach for the development of a CE method for OME impurity profiling. The scouting experiments suggested the selection of solvent modified Micellar ElectroKinetic Chromatography operative mode using a pseudostationary phase composed of sodium dodecyl sulfate (SDS) micelles and n-butanol as organic modifier in borate buffer. A symmetric three-level screening matrix 37//16 was used to evaluate the effect of Critical Method Parameters, including Background Electrolyte composition and instrumental settings, on Critical Method Attributes (critical resolution values, OME peak width and analysis time). The analytical procedure was optimized using Response Surface Methodology through a Central Composite Orthogonal Design. Risk of failure maps made it possible to define the Method Operable Design Region, within which the following optimized conditions were selected: 72â¯mM borate buffer pH 10.0, 96â¯mM SDS, 1.45â¯%v/v n-butanol, capillary temperature 21 °C, applied voltage 25â¯kV. The method was validated according to ICH guidelines and robustness was evaluated using a Plackett-Burman design. The developed procedure enables the simultaneous determination of OME and seven related impurities, and has been successfully applied to the analysis of pharmaceutical formulations.
Asunto(s)
Contaminación de Medicamentos , Electroforesis Capilar , Omeprazol , Inhibidores de la Bomba de Protones , Dodecil Sulfato de Sodio , Omeprazol/análisis , Omeprazol/química , Contaminación de Medicamentos/prevención & control , Electroforesis Capilar/métodos , Inhibidores de la Bomba de Protones/análisis , Dodecil Sulfato de Sodio/química , Reproducibilidad de los Resultados , Solventes/química , Cromatografía Capilar Electrocinética Micelar/métodos , Concentración de Iones de Hidrógeno , Micelas , 1-Butanol/químicaRESUMEN
Genetic factors, diet, lifestyle, and other factors lead to various complications in the body, such as obesity and other chronic diseases. The inflammatory state caused by excessive accumulation of body fat affects the pathways related to the control of glycemic homeostasis, leading to a high demand for insulin, to subsequent failure of stressed ß cells, and development of type 2 diabetes mellitus (T2DM). The study of new endocrine signalers, such as bile acids (BAs), becomes necessary as it allows the development of alternatives for T2DM treatment. In this work, a methodology was developed to quantify tauroursodeoxycholic BA (TUDCA) in liver cells of the HepG2 strain treated in hyperlipidic medium. This BA helps to improve insulin clearance by increasing the expression of the insulin-degrading enzyme, restoring sensitivity to this hormone, and making it viable for treating T2DM. Herein, a targeted metabolomic method for TUDCA determination in extracellular medium of hepatocyte matrices by micellar electrokinetic chromatography-UV was optimized, validated, and applied. The optimized background electrolyte was composed of 40 mmol/L sodium cholate and 30 mmol/L sodium tetraborate at pH 9.0. The following figures of merit were evaluated: linearity, limit of quantification, limit of detection, accuracy, and precision. Data obtained with the validated electrophoretic method showed a self-stimulation of TUDCA production in media supplemented only with BA. On the other hand, TUDCA concentration was reduced in the hyperlipidic medium. This suggests that, in these media, the effect of TUDCA is reduced, such as self-stimulated production and consequent regulation of glycemic homeostasis. Therefore, the results reinforce the need for investigating TUDCA as a potential T2DM biomarker as well as its use to treat several comorbidities, such as obesity and diabetes mellitus.
Asunto(s)
Cromatografía Capilar Electrocinética Micelar , Diabetes Mellitus Tipo 2 , Obesidad , Ácido Tauroquenodesoxicólico , Ácido Tauroquenodesoxicólico/farmacología , Ácido Tauroquenodesoxicólico/análisis , Ácido Tauroquenodesoxicólico/metabolismo , Humanos , Obesidad/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Células Hep G2 , Cromatografía Capilar Electrocinética Micelar/métodos , Hepatocitos/metabolismo , Hepatocitos/efectos de los fármacos , Reproducibilidad de los Resultados , Metabolómica/métodos , Modelos Lineales , Límite de DetecciónRESUMEN
INTRODUCTION: Gomisin is a natural dibenzo cyclooctene lignan, which is mainly derived from the family Magnoliaceae. It has anti-inflammatory, antioxidant, anti-tumor, anti-aging, and hypoglycemic effects. Gomisins play important roles as medicines, nutraceuticals, food additives, and cosmetics. OBJECTIVE: The objective of this study is to establish a micellar electrokinetic chromatography (MEKC) method for simultaneous separation and determination of seven biphenyl cyclooctene lignans (Gomisin D, E, G, H, J, N, and O) in Schisandra chinensis and its preparations. METHODS: The method was optimized by studying the effects of the main parameters on the separation. The method has been validated and successfully applied to the determination of seven Gomisins in S. chinensis and its preparations. RESULTS: In the separation system, the running buffer was composed of 20 mM Na2HPO4, 8.0 mM sodium dodecyl sulfate (SDS), 11% (v/v) methanol, and 6.0% (v/v) ethanol. A diode array detector was used with a detection wavelength of 230 nm, a separation voltage of 17 kV, and an operating temperature of 25°C. Under this condition, the seven analytes were separated at baseline within 20 min, and a good linear relationship was obtained with correlation coefficient ranging from 0.9919 to 0.9992. The limit of detection (LOD, S/N = 3) and the limit of quantification (LOQ, S/N = 10) ranged from 0.8 to 0.9 µg/mL and from 2.6 to 3.0 µg/mL, respectively. The recovery rate was between 99.1% and 102.5%. CONCLUSION: The experimental results indicated that this method is suitable for the separation and determination of seven Schisandra biphenyl cyclooctene lignan compounds in real samples. At the same time, it provides an effective reference for the quality control of S. chinensis and its preparations.
Asunto(s)
Cromatografía Capilar Electrocinética Micelar , Ciclooctanos , Lignanos , Schisandra , Solventes , Lignanos/análisis , Schisandra/química , Cromatografía Capilar Electrocinética Micelar/métodos , Solventes/química , Ciclooctanos/análisis , Ciclooctanos/química , Reproducibilidad de los Resultados , Límite de Detección , Compuestos de Bifenilo/químicaRESUMEN
The two-step preconcentration technique consisting of large-volume sample stacking (LVSS) and micelle to solvent stacking (MSS) in cyclodextrin-modified electrokinetic chromatography (CDEKC) was developed for the analysis of five cationic alkaloids in complex Chinese herbal prescriptions. Relevant parameters affecting separation and stacking performance were optimized separately. Under the optimal LVSS-MSS-CDEKC conditions, less analysis time and organic solvent were required, and the enhancement factors of analytes ranged from 12 to 15 compared with the normal CDEKC separation mode. Further, all validation results demonstrated good applicability and multiple alkaloids (epiberberine, dehydrocorydaline, jatrorrhizine, coptisine and berberine) in Yangxinshi tablet (YXST) have been simultaneously determined. This approach presents powerful potential for the determination of multiple components in complex preparations of Chinese medicine.
Asunto(s)
Alcaloides , Cromatografía Capilar Electrocinética Micelar , Medicamentos Herbarios Chinos , Comprimidos , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/análisis , Cromatografía Capilar Electrocinética Micelar/métodos , Comprimidos/química , Alcaloides/análisis , Alcaloides/química , Reproducibilidad de los Resultados , Micelas , Modelos Lineales , Ciclodextrinas/química , Límite de DetecciónRESUMEN
OBJECTIVES: The Making Every Contact Count (MECC) initiative is broadly defined as an opportunistic approach to prevention by making use of the thousands of conversations service providers have with service users every day. However, since its conception, the application of MECC has diverged and developed considerably. Thus, the current study aimed to revise the definition according to current research and practice to better describe what is and is not included. STUDY DESIGN: A consensus building classic Delphi methodology, completed by an expert panel. METHODS: Round 1 asked open questions around the definition of MECC. Content analysis of round 1 identified statements that were rated for agreement in round 2. Statements achieving ≥80% agreement were included in a short, long, or operational definition of MECC that were rated for agreement in round 3 (the minimum number required). An agreement of ≥80% indicated consensus. RESULTS: Forty out of 100 contacted experts completed three rounds. Experts in practice and research were recruited internationally although most were from England. From round 1, 274 statements were generated, of which 96 achieved consensus and were included within round 3. The short and long definition received consensus in round 3, the operational definition required four rounds to reach consensus. CONCLUSIONS: MECC is a person-centred approach to health behaviour change that, provided an individual possesses the relevant skills, can be delivered by anyone and anywhere. The distinguishing feature of MECC is not in its duration, target behaviour, or conditions for delivery, but rather in the approach taken and the mechanisms applied to conversations. Implications for research and practice are discussed, and the limits for applicability acknowledged.
Asunto(s)
Cromatografía Capilar Electrocinética Micelar , Humanos , Consenso , Técnica Delphi , Proyectos de Investigación , InglaterraRESUMEN
INTRODUCTION: Yangxinshi tablet (YXST) is a traditional Chinese medicine preparation characterized by its high efficacy and safety for the treatment of cardiovascular diseases. Anionic compounds have been revealed as potential active components. However, there is currently limited research regarding its quality control. OBJECTIVE: We aimed to establish a strategy for the simultaneous separation and determination of five key anionic compounds in YXST. METHOD: A sensitive and efficient analytical method was developed and applied for the simultaneous separation and determination of five key compounds in YXST using large-volume sample stacking with polarity switching and micelle electrokinetic chromatography (LVSS-PS-MEKC) coupled with diode array detection. Crucial parameters, including sample volume, applied voltage, composition and pH of the running buffer, concentration of organic modifier, and switching time of the polarity, were systematically evaluated and optimized using a single variable method to enhance separation performance. Furthermore, the impact of cyclodextrin and sodium dodecyl sulfate as electrolyte modifiers was also investigated. RESULTS: Under the optimal conditions, baseline separation of the five compounds (daidzein, puerarin, glycyrrhiztinic acid, chlorogenic acid, and salvianolic acid B) was achieved within 20 min. In comparison to the conventional MEKC mode, the constructed LVSS-PS-MEKC method exhibited a more than sixfold increase in the enrichment factor. The method was validated in terms of linearity, precision, accuracy, 24 h stability, and recovery and successfully applied to analyze YXST samples. CONCLUSION: A sensitive strategy was developed for the simultaneous separation and determination of five key anionic components in YXST, offering a robust and efficient strategy for pharmaceutical analysis.
Asunto(s)
Aniones , Cromatografía Capilar Electrocinética Micelar , Medicamentos Herbarios Chinos , Comprimidos , Cromatografía Capilar Electrocinética Micelar/métodos , Comprimidos/química , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/análisis , Reproducibilidad de los Resultados , Dodecil Sulfato de Sodio/químicaRESUMEN
Glutamine is the most abundant free proteinogenic α-amino acid. It is naturally produced in the organism and acts as a precursor for the synthesis of different biologically important molecules (such as proteins or nucleotides). However, under stressful conditions, the organism is unable to produce it in enough amounts to function properly. Thus, glutamine (Gln)-based supplements have become increasingly popular over the last decade. Since legal regulations establish that amino acid-based dietary supplements must contain only the L-enantiomer and not the racemate, adequate chiral methodologies are required to achieve their quality control. In this work, an analytical methodology based on the use of micellar electrokinetic chromatography is proposed for the rapid enantiomeric determination of DL-Gln in dietary supplements. Using (+)-1-(9-fluorenyl)-ethyl chloroformate as a derivatizing agent and ammonium perfluorooctanoate as separation medium, the Gln diastereoisomers formed under optimal conditions were separated in 8 min with a resolution of 2.8. The analytical characteristics of the method were evaluated in terms of linearity, precision, accuracy, and limits of detection/quantitation, and they were found appropriate for the analysis of L-Gln-based dietary supplements.
Asunto(s)
Cromatografía Capilar Electrocinética Micelar , Glutamina , Cromatografía/métodos , Aminoácidos/química , Suplementos Dietéticos/análisis , Estereoisomerismo , Cromatografía Capilar Electrocinética Micelar/métodosRESUMEN
The first chiral methodology enabling the separation of indacaterol enantiomers was developed in this work by cyclodextrin-electrokinetic chromatography. Indacaterol (IND) is a chiral drug marketed as a pure enantiomer. Then, the separation and quantification of each enantiomer is of great importance for the quality control of pharmaceutical formulations. After selecting the most suitable chiral selector and background electrolyte, two Box-Behnken designs were achieved to optimize the electrophoretic conditions using two different approaches to shorten analysis times: i) decreasing the capillary length, or ii) performing a short-end injection. Indacaterol enantiomers were separated in less than 5 min with a resolution value of 3.6 under the optimal separation conditions: 0.7% (m/v) carboxymethyl-α-cyclodextrin in 50 mM sodium formate buffer (pH 4.0) and using a short-end injection. Then, the analytical characteristics of the method were evaluated and LODs of 0.05 mg/L for S-IND and 0.04 mg/L for R-IND were achieved. Also, the method allowed the detection of a 0.1% enantiomeric impurity (S-IND) in the R-IND-based pharmaceutical formulations. The developed method was applied to the analysis of two pharmaceutical formulations. Percentages of 97 ± 3% and 103 ± 6% of R-IND with respect to the labeled amounts were found.
Asunto(s)
Cromatografía Capilar Electrocinética Micelar , Ciclodextrinas , Indanos , Quinolonas , Cromatografía , Ciclodextrinas/química , Preparaciones Farmacéuticas , Estereoisomerismo , Cromatografía Capilar Electrocinética Micelar/métodosRESUMEN
Propranolol is currently considered as an emerging contaminant in water bodies. In this study, R- and S-propranolol were determined in river samples by electrokinetic chromatography (EKC) using nanodiamonds (NDs) and human serum albumin (HSA) as a pseudo-stationary phase in order to achieve enantioseparation. Previously, river samples were preconcentrated using a column filled with Amberlite® IR-120 and Dowex® 50WX8 resins. The setting up of influential factors such as temperature, voltage, pH, and HSA and NDs concentration is accurately described along this manuscript. A multivariate study and optimization was carried out to obtain the enantioseparation of propranolol (Rs = 2.91), which was reached under the following experimental conditions: voltage of 16 kV, temperature of 16°C, phosphate buffer pH 9.5, NDs of 0.20%, and HSA of 15 µmol l-1 . The recoveries of analytes under optimal conditions were higher than 98%. The limits of detection were 0.85 µg l-1 for R- and S-propranolol. The method was applied to real samples, and the obtained results in three different water sources studied were 1.02, 0.59, and 0.30 µg l-1 for the R-enantiomer and 0.99, 0.54, and 0.28 µg l-1 for the S-enantiomer. The accuracy of the proposed methodology (including bias and precision) has allowed us to propose it as a successful tool for the control of water quality.
Asunto(s)
Cromatografía Capilar Electrocinética Micelar , Nanodiamantes , Humanos , Propranolol , Albúmina Sérica Humana , Ríos , Estereoisomerismo , Cromatografía Capilar Electrocinética Micelar/métodosRESUMEN
Herein, we have developed a novel method of aqueous-sample dispersive liquid-liquid microextraction (AqS-DLLME) followed by sweeping micellar electrokinetic chromatography-tandem mass spectrometry (MEKC-MS/MS) for simultaneous determination of breast cancer drugs letrozole, anastrozole, palbociclib, ribociclib, abemaciclib, and fulvestrant in human plasma. Coupling of MEKC to MS was possible due to the use of ammonium perfluorooctanoate (APFO) as a volatile surfactant. The MEKC and MS conditions were optimized to achieve a fast, sensitive, selective, and green analysis enabling full separation of the analytes within 16 min. Electrophoretic buffer was 125 mM APFO at apparent pH 10.5 in 32 % MeOH, while sheath liquid was 70 % MeOH with 0.2 % formic acid, delivered at 10 µL/min. Excellent extraction recoveries from plasma ranging from 89.4 to 104.9 % were obtained with a combination of protein precipitation and DLLME. The developed method was validated according to the ICH guidelines. Remarkable selectivity, accuracy (bias < 6.7 %), precision (RSD < 15.8 %), and stability (bias < 10.4 %) with insignificant matrix effect (RSD < 14.0 %) and no carry-over were obtained over a wide range of concentrations. Linearity with inter-day slope RSD lower than 8.7 % was demonstrated. With this method, very low concentrations could be detected after the injection of only 68.7 nL of the sample. The method was applied to plasma samples from six women currently receiving breast cancer treatment. Determined concentrations of the drugs of interest agreed with concentrations found in clinical studies, thus proving the suitability of the developed method for therapeutic drug monitoring as a superior alternative to published LC-MS methods.
Asunto(s)
Neoplasias de la Mama , Caprilatos , Cromatografía Capilar Electrocinética Micelar , Fluorocarburos , Microextracción en Fase Líquida , Humanos , Femenino , Espectrometría de Masas en Tándem , Neoplasias de la Mama/tratamiento farmacológico , Cromatografía Capilar Electrocinética Micelar/métodos , Microextracción en Fase Líquida/métodos , MicelasRESUMEN
In this work, a capillary electrophoresis method was developed as a quality control tool to determine the enantiomeric purity of a series of five chiral compounds evaluated as potential severe acute respiratory syndrome coronavirus 2 3CL protease inhibitors. The first cyclodextrin tested, that is, highly sulfated-ß-cyclodextrin, at 6% (m/v) in a 25 mM phosphate buffer, using a capillary dynamically coated with polyethylene oxide, at an applied voltage of 15 kV and a temperature of 25°C, was found to successfully separate the five derivatives. The limits of detection and quantification were calculated together with the greenness score of the method in order to evaluate the method in terms of analytical and environmental performance. In addition, it is noteworthy that simultaneously high-performance liquid chromatography separation of the enantiomers of the same compounds with two different columns, the amylose tris(3,5-dimethylphenylcarbamate)-coated and the cellulose tris(3,5-dichlorophenylcarbamate)-immobilized on silica stationary phases, was studied. Neither the former stationary phase nor the latter was able to separate all derivatives in a mobile phase consisting of n-heptane/propan-2-ol 80/20 (v/v).
Asunto(s)
SARS-CoV-2 , Estereoisomerismo , Inhibidores de Proteasas/aislamiento & purificación , Inhibidores de Proteasas/química , Inhibidores de Proteasas/farmacología , Inhibidores de Proteasas/análisis , Proteasas 3C de Coronavirus/química , Proteasas 3C de Coronavirus/antagonistas & inhibidores , Cromatografía Capilar Electrocinética Micelar/métodos , Límite de Detección , COVID-19 , Humanos , Betacoronavirus/aislamiento & purificación , Betacoronavirus/química , Cromatografía Líquida de Alta Presión/métodosRESUMEN
An on-line enrichment and separation of multiple derivatized monosaccharides with cyclodextrin-encapsulated sweeping (CDES) by micellar electrokinetic chromatography (MEKC) was presented. Five monosaccharides (L-(-)-Mannose, D-(+)-Glucose, D-(-)-Ribose, D-(+)-Xylose, and L-(+)-Rhamnose) were derivatized with 1-phenyl-3-methyl-5-pyrazolone, subsequently concentrated and separated by MEKC. The optimized conditions were as follows: 50 mM phosphoric acid (PA), 100 mM sodium dodecyl sulfate (SDS), and 30 % (v/v) methanol in background solution; 140 s injection of sample solution containing 50 mM CD and 100 mM PA, followed by 90 s injection of 40 mM SDS solution. Under the optimized conditions, the correlation coefficients ≥ 0.9953, and the limits of detection ranged from 4.2 to 7.4 ng/mL. Relative standard deviation values ranged from 0.24-4.23 %, and sensitivity enrichment factors were in the range of 53-82 compared with typical injection (50 mbar, 3 s). The CDES-MEKC method was successfully applied to Jujube with good recoveries of 84.22-104.33 %. The method provides new ideas for the on-line enrichment and detection of trace monosaccharides and even other target analytes in foods with complex matrices.
Asunto(s)
Cromatografía Capilar Electrocinética Micelar , Ciclodextrinas , Cromatografía Capilar Electrocinética Micelar/métodos , Ciclodextrinas/química , Monosacáridos , Frutas , MicelasRESUMEN
A method using micellar electrokinetic chromatography coupled with large-volume sample stacking for the determination of ticagrelol was developed and validated. The analysis was performed in a fused silica capillary (41.5 cm effective length, 50 µm diameter) with ultraviolet detection at 195 nm. The background electrolytes were 30 mM phosphate buffer of pH 3.0 with 120 mM sodium dodecylsulfate and 10 % (v/v) acetonitrile (120 s X 50 mbar; 20°C; -18 kV) and 30 mM borate buffer of pH 8.5 with 75 mM sodium dodecylsulfate (120 s X 50 mbar; 20°C; 25 kV); under acidic and alkaline conditions, respectively. The method was found to be reliable with respect to specificity, linearity of the calibration line (R2 > 0.99), repeatability (relative standard deviation 2.56%-3.34%), and accuracy (recovery in the range 101.21%-102.67%). The limits of detection and quantitation were 0.032, 0.071, and 0.087, 0.188 µg/mL, respectively. The method was successfully applied for the determination of ticagrelol concentrations in rat plasma and tablets with good recoveries and reproducibility. The presented method proved to be suitable for monitoring ticagrelor in rat plasma.
Asunto(s)
Cromatografía Capilar Electrocinética Micelar , Ratas , Animales , Cromatografía Capilar Electrocinética Micelar/métodos , Micelas , Reproducibilidad de los Resultados , Comprimidos , SodioRESUMEN
A novel online two-step pressure injection-assisted stacking preconcentration method, which involves sweeping and affinity micelles in micellar electrokinetic chromatography was developed to simultaneously measure various organic anions. The micellar solution was a mixed solution that contained 0.3 mM didodecyldimethylammonium bromide and 20 mM borax. After the micellar solution was injected for 60 s, the tested analytes prepared in 20 mM borax were introduced into the capillary for 150 s. The key experimental factors that influenced the separation and sensitivity were investigated and optimized, including the concentration and injection time of the micellar solution, the concentration of borax in the sample solution, the concentration of sodium dodecyl sulfate and borax in the background electrolyte (BGE), the content of acetonitrile in the BGE and the injection time of the sample solution. Compared with typical injection methods, this method achieved sensitivity enhancement factors ranging from 85 to 97 under optimized conditions. Good linearity for matrix-matched calibration was established for all analytes with R2 values of 0.9986-0.9996. The intraday (n = 6) and interday (n = 6) precisions of the method were less than 2.85% when expressed as relative standard deviations. When the method was applied to analyze rice and dried ginger samples, analyte recoveries ranged from 85.81% to 106.59%. Through sweeping and affinity micelles, stacking preconcentration method was successfully employed to analyze trace amounts of fenoprop and 2,4-dichlorophenoxyacetic acid in rice and dried ginger samples.
Asunto(s)
Cromatografía Capilar Electrocinética Micelar , Herbicidas , Herbicidas/análisis , Cromatografía Capilar Electrocinética Micelar/métodos , Micelas , AnionesRESUMEN
High-performance layer electrochromatography (HPLEC) combines the advantages of overpressured-layer chromatography (OPLC) and pressurized planar electrochromatography (PPEC) while overcoming some of their limitations. HPLEC equipment can work in various HPLEC, OPLC, and PPEC modes. The equipment enables HPLEC analysis also with an electroosmotic effect directed against the hydrodynamic flow of the mobile phase. The change in the electric field direction in the separation system does not result in a change in either the direction of the mobile phase flow or the direction of solute migration. The hydrodynamic flow generated by the pump dominates the electroosmotic effect and enables separation against the direction of the latter. Reversed-polarization HPLEC may be advantageous for the analysis of anionic compounds, as it facilitates faster and more selective separation than OPLC performed in similar conditions. This separation mode provides a new possibility to develop and optimize separation methods by performing separation against the electroosmotic effect and without need of any modification of the adsorbent surface. A drawback of this separation mode is the increase in the backpressure at the mobile phase inlet and the limitation of the mobile phase flow rate. Currently, contrary to the single-channel mode, multi-channel reverse-polarity HPLEC still requires some technical and methodological improvements.
Asunto(s)
Cromatografía Capilar Electrocinética Micelar , Cromatografía en Capa Delgada/métodos , Cromatografía Líquida de Alta Presión/métodos , Indicadores y Reactivos , Cromatografía Capilar Electrocinética Micelar/métodosRESUMEN
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-ß-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-ß-cyclodextrin; temperature, 22 °C; voltage, 23-29 kV. The method was validated and applied to ampoules drug products.
Asunto(s)
Cromatografía Capilar Electrocinética Micelar , Ciclodextrinas , Ciclodextrinas/química , Micelas , Cromatografía Capilar Electrocinética Micelar/métodos , Trimecaína , Dodecil Sulfato de Sodio , Boratos , Reproducibilidad de los ResultadosRESUMEN
An in-capillary 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate) as oxyradicals combining field-enhanced sample injection with micellar electrokinetic chromatography was developed for screening and determination of the major antioxidants in Yangxinshi Tablet. To obtain simultaneous separation and detection of radicals and analytes, relevant factors were optimized separately. Under the optimum conditions, four compounds including salvianolic acid B, hyperoside, puerarin, and caffeic acid were identified as the major antioxidants. All validation results covering recovery, precision, and stability demonstrated good applicability of the method. On this basis, the total antioxidant activity was successfully evaluated in terms of the decreased peak area of radicals. There was a correlation coefficient of 0.8974 between the total contents of major antioxidants and the total antioxidative activity of the sample. Therefore, these four compounds were selected as combinatorial markers for the quality evaluation of Yangxinshi Tablet. It was concluded that the established method presented a powerful potential to screen and quantify active ingredients in the complex preparation of Chinese medicine.
Asunto(s)
Cromatografía Capilar Electrocinética Micelar , Medicamentos Herbarios Chinos , Antioxidantes/análisis , Micelas , Medicamentos Herbarios Chinos/análisis , Cromatografía Capilar Electrocinética Micelar/métodosRESUMEN
Recently, non-steroidal anti-inflammatory drugs (NSAIDs) have been increasingly used in humans and animals. Despite being effective against a wide variety of diseases, they pose a threat to aquatic environments. In the current work, a highly efficient, selective, and sensitive micellar electrokinetic chromatography (MEKC) method was developed for the determination of five NSAIDs in environmental water samples. The optimal separation BGE was 15 mM borate buffer (pH 9), 90 mM SDS, and 10% methanol at a separation voltage of 15 kV and a hydrodynamic injection of 10 mbar for 5 s. The results presented in this study provide a higher number of theoretical plates N > 780 000 with excellent RSDs of 0.1-1.5% and great sensitivity (3-15 µg L-1) for NSAIDs. To validate this method, the solid phase extraction method was optimized using two different cartridges (C18 and Oasis HLB); the results showed excellent recoveries (73-111.6%) for all the analytes in wastewater samples.
Asunto(s)
Cromatografía Capilar Electrocinética Micelar , Aguas Residuales , Humanos , Animales , Cromatografía Capilar Electrocinética Micelar/métodos , Extracción en Fase Sólida , Antiinflamatorios no EsteroideosRESUMEN
Ammonium perfluorooctanoate (APFOA) was used as a surfactant for the separation of free unsaturated C18 fatty acids by micellar electrokinetic chromatography. A simple background electrolyte of 50 mM APFOA water/methanol (90:10, v/v) at pH = 10 enabled the repeatable separation of oleic acid, elaidic acid, linoleic acid, and alpha-linolenic acid in less than 20 min. Separation conditions were optimized regarding various parameters (organic solvent, counterion, APFOA concentration, and pH). Because the repulsive interactions between fluorocarbon chains and hydrogenated chains are known to lead to segregation and phase separation, the choice of perfluorinated micelles to separate such perhydrogenated long-chain acids could appear astonishing. Therefore, the critical micelle concentration, the charge density, and the mobility of the micelles have been determined, resulting in a first description of the separation process.
Asunto(s)
Cromatografía Capilar Electrocinética Micelar , Micelas , Cromatografía Capilar Electrocinética Micelar/métodos , Tensoactivos/química , CaprilatosRESUMEN
Realising the need to devise a simple, sensitive, and reliable detection method, this study investigated the development of a dual-stacking transient isotachophoresis (t-ITP) and sweeping in micellar electrokinetic chromatography with diode array detector (t-ITP/sweeping-MEKC-DAD) for the determination of selected non-steroidal anti-inflammatory drugs (NSAIDs); ketoprofen, diclofenac and naproxen from aqueous matrices. Prior to the system setup, various parameters were optimised to assess the potential use of the t-ITP paired with the sweeping stacking technique in micellar background electrolyte for dual preconcentration and separation of trace amounts of NSAIDs. Once the optimum conditions have been established, the method performance was validated and applied to 17 environmental water samples. Based on the results, the combined t-ITP and sweeping approach significantly improved the stacking and separation sensitivity. A large volume of samples could also be introduced and subsequently separated by MEKC with greater focusing effects due to the sweeping. Under optimised conditions, the developed method exhibited excellent linearity at a high range (0.1-500 ng/mL, r2 ≥ 0.998), low limits of detection (LODs) of 0.01-0.07 ng/mL, and a remarkable relative recovery (RR) of 99.6-101.9% with a relative standard deviation (RSD) of 1.4-8.6% (n = 9). Ultimately, the sensitivity enhancement factors improved up to 666-fold using the optimised method. Therefore, the proposed method presents a simplified yet effective and suitable for the determination of NSAIDs from aqueous matrices.