Microdevice for separation and quantitative fraction collection

A new microfluidic concept for quantitative whole-column fraction collection of electrophoretically separated zones was developed. The prototype device, fabricated on a polycarbonate disk by injection molding, integrated electrophoretic separation channels with fraction collection reservoirs distributed along the separation channel. The microdevice was designed in a CD-like format to use the centrifugal force for moving the liquid in the microchannels. A serpentine shape of the separation channel was selected to create segments for quantitative whole-column fraction collection. The operation was tested with visual monitoring of isotachophoretic separation and collection of cationic dyes.     

Investigation of the separability of thaumatin by capillary electrophoresis

The electrophoretic behaviour of the highly basic protein thaumatin was explored in strongly acid (pH 2) and mildly acid (pH 4.5) separation systems using both bare and coated fused silica capillaries. The separation selectivity for thaumatin I, thaumatin II, and for other sample constituents was insufficient for their baseline separation at pH 2 in an uncoated capillary because the separation efficiency was markedly lower than is common in the electrophoretic separations of proteins. A separation selectivity higher by up to one order of magnitude has been reached at pH 4.5. A pronounced asymmetry of zones, which impaired resolution at this pH, was effectively suppressed by coating of the capillary wall with a polymer. In fact, adsorption on the capillary coating always plays a contributory role whenever a good separation of thaumatin constituents is attained. This indicates that electrochromatographic separation systems based on capillaries coated with the layer of either cationic or hydrophilic uncharged polymer hold promise for the development of methods for thaumatin analysis.     

Electrochemical determination of hesperidin using mesoporous SiO2 modified electrode

A simple micro-capillary electrophoresis system to be used as disposable device was developed. A short commercial capillary was used as the separation channel, hydrostatic pressure generated by the sample employed for injection, and a voltage of 200 V used for separation in a 6 cm long capillary assisted by hydrostatic pressure of the carrier. The device was used for the separation of dopamine and catechol. Good reproducibility and efficiency was obtained. Because the instrumentation and operation conditions were simplified, and a replaceable modular separation channel was used, the proposed micro-capillary electrophoresis system is potentially useful in disposable devices.     

High-performance liquid chromatographic separation of subcomponents of antimycin A

Using a reversed-phase high-performance liquid chromatographic (HPLC) technique, a mixture of antimycins A was separated into eight hitherto unreported subcomponents, A1a, A1b, A2a, A2b, A3a, A3b, A4a, and A4b. Although a base-line resolution of the known four major antimycins A1, A2, A3, and A4 was readily achieved with mobile phases containing acetate buffers, the separation of the new antibiotic subcomponents was highly sensitive to variation in mobile phase conditions. The type and composition of organic modifers, the nature of buffer salts, and the concentration of added electrolytes had profound effects on capacity factors, separation factors, and peak resolution values. Of the numerous chromatographic systems examined, a mobile phase consisting of methanol-water (70:30) and 0.005 M tetrabutylammonium phosphate at pH 3.0 yielded the most satisfactory results for the separation of the subcomponents. Reversed-phase gradient HPLC separation of the dansylated or methylated antibiotic compounds produced superior chromatographic characteristics and the presence of added electrolytes was not a critical factor for achieving separation. Differences in the chromatographic outcome between homologous and structural isomers were interpreted based on a differential solvophobic interaction rationale. Preparative reversed-phase HPLC under optimal conditions enabled isolation of pure samples of the methylated antimycin subcomponents for use in structural studies.     

Robust and simple interface for microchip electrophoresis-mass spectrometry

A robust and simple interface for microchip electrophoresis-mass spectrometry (MCE-MS) was developed using a spray nozzle connected to the exit of the separation channel of the microchip. The spray nozzle was attached to the microchip using a polyether ether ketone screw without adhesive, thus allowing easy replaced. Sample injection and electrophoretic separation was performed by control of the voltage only. The analysis of a few basic drugs was performed using the optimized MCE-MS system. The separation was improved by using a high-viscosity separation buffer and a spray nozzle with a small bore size. This system was also applied to the separation of peptides and protein-trypsin digests. Sample adsorption was minimized by adding acetonitrile to the separation buffer when using a quartz microchip.     

Electrochromatographic evaluation of a silica monolith capillary column for separation of basic pharmaceuticals

A silica-based monolithic capillary column was prepared via a sol-gel process. The continuous skeleton and large through-pore structure were characterized by scanning electron microscopy (SEM). The native silica monolith has been successfully employed in the electrochromatographic separation of beta-blockers and alkaloids extracted from traditional Chinese medicines (TCMs). Column efficiencies greater than 250 000 plates/m for capillary electrochromatography (CEC) separation of basic compounds were obtained. It was observed that retention of basic pharmaceuticals on the silica monolith was mainly contributed by a cation-exchange mechanism. Other retention mechanisms including reversed-phase and normal-phase mechanisms and electrophoresis of basic compounds also played a role in separation. A comparison of the differences between CEC and capillary zone electrophoresis (CZE) separation was also discussed.     

微流控芯片上同工酶的孵育及活性检测

建立了一种在微流控芯片上进行同工酶孵育及活性检测的方法. 该方法在集成温控装置的微流控芯片上实现对同工酶与辅酶反应进程的控制, 完成同工酶的进样、孵育反应、电泳分离和活性检测的实验步骤. 建立了基于微流控芯片的同工酶荧光检测系统, 使用360 nm光源激发辅酶产生荧光, 在460 nm处选择性采集荧光信号. 在微流控芯片上实现了同工酶样品的快速活性检测, 酶活性检测限达到0.5 U/L.     

Optimization of high-performance liquid chromatographic parameters for the determination of capsaicinoid compounds using the simplex method.

A high-performance liquid chromatographic method was developed for the analysis of capsaicinoid compounds, the pungent principles of capsicum fruits. A sequential simplex method was applied to optimize the chromatographic response function used to assess the quality of separation by varying the chromatographic parameters. The separation was achieved in 11 min using a C-8 column of 15-cm length and 4.6 mm diameter using a UV detector. A flow rate of 1.15 ml min(-1) at a column temperature of 43.5 degrees C using 63.7% methanol in water gave the most efficient separation. The method was found to be suitable for the determination of the major capsaicinoid compounds in the capsicum samples.     

以L-天冬氨酸为手性选择剂对手性药物的毛细管电泳拆分

以Ｌ－天冬氨酸为手性选择剂,建立了对１０种手性药物旋光异构体的毛细管电泳快速分离方法,优化了分离条件,并对分离机理作了初步探讨。     

Open tubular capillary column of 50 μm internal diameter with a very high separation efficiency for the separation of peptides in CEC and LC

A specially designed long open tubular capillary column (50 μm internal diameter and 112 cm effective length) was prepared by fabrication of a thin three‐component co‐polymer layer on the inner surface of silica capillary. A pretreated silica capillary was reacted with 4‐(chloromethyl)phenyl isocyanate in the presence of dibutyltin dichloride as catalyst followed by sodium diethyl dithiocarbamate. Then a thin polymer layer was made on the inner surface of capillary by reversible addition‐fragmentation transfer polymerization of styrene, N‐phenylacrylamide, and methacrylic acid. A carefully adjusted formulation of reaction mixture and elaborated procedures were adopted to secure formation of the co‐polymer layer of enhanced separation performance. The co‐polymer immobilized open tubular capillary column was used for the separation of a synthetic mixture of five peptides and excellent separation efficiency (over 1.7 million per column) was obtained in the capillary electrochromatography mode. Such excellent separation efficiencies of ca. 1 m column have not been obtained in the isocratic elution mode so far. The column was also used for separation of the peptides in the liquid chromatography mode to show very good separation efficiency (average 286 700 per column).     

Determination of inosine 5′-monophosphate and guanosine 5′-monophosphate in pig feed by capillary zone electrophoresis

A capillary zone clectrophoresis method was developed for the determination of IMP and GIMP, commonly used as flavor enhancers in poultry feed, in a real sample of complex composition. A baseline separation of inosine 5′-monophosphate and guanosine 5′-monophosphate was achieved within 10 min and the other components in the sample did not interfere with the separation. Quantitative results obtained from pig feed samples are presented. The separation conditions and experimental reproducibility are also discussed.     

Hot embossing of electrophoresis microchannels in PMMA substrates using electric heating wires

A simple method based on electric heating wires has been developed for the rapid fabrication of poly(methyl methacrylate) (PMMA) electrophoresis microchips in ordinary laboratories without the need for microfabrication facilities. A piece of stretched electric heating wire placed across the length of a PMMA plate along its midline was sandwiched between two microscope slides under pressure. Subsequently, alternating current was allowed to pass through the wire to generate heat to emboss a separation microchannel on the PMMA separation channel plate at room temperature. The injection channel was fabricated using the same procedure on a PMMA sheet that was perpendicular to the separation channel. The complete microchip was obtained by bonding the separation channel plate to the injection channel sheet, sealing the channels inside. The electric heating wires used in this work not only generated heat; they also served as templates for embossing the microchannels. The prepared microfluidic microchips have been successfully employed in the electrophoresis separation and detection of ions in connection with contactless conductivity detection.     

Forced-flow chromatography of the lanthanides with continuous in-stream detection

A method for ion-exchange separation of rare earths is described in which the elements are detected automatically after addition of a colour-forming reagent. A separation of 13 rare-earth elements was accomplished in 4 hr.     

1 μm硅胶颗粒固定相的加压电色谱分离性能

制备了1 μm无孔硅胶颗粒。通过电动填充法得到总长度为45 cm(固定相填充长度为20 cm)、内径为100 μm的毛细管色谱柱。以乙腈-水体系作为流动相,详细考察了碱性化合物在该色谱柱上的加压电色谱(pCEC)分离性能,讨论了流动相比例、缓冲液浓度、pH值及操作电压等因素对分离的影响。实验结果表明,裸硅胶柱在乙腈-水体系分离碱性样品中表现出典型的反相色谱分离性能;缓冲液浓度的改变则对分离影响不大。当pH值改变时,碱性化合物的解离程度发生变化,它们与固定相之间的作用力发生变化,使得分离度发生相应的变化。分离柱效随施加电压的增加而增加,在1 kV电压下,裸硅胶柱对邻甲苯胺的柱效为35000理论塔板/m。     

A lab-on-a-chip for rapid blood separation and quantification of hematocrit and serum analytes

In this work, a new lab-on-a-chip for rapid analysis of low volume blood samples was designed, fabricated and demonstrated for integration of serum separation, hematocrit evaluation, and protein quantitation. Blood separation was achieved using microchannel flow-based separation. A novel method for evaluating hematocrit from microfluidic flow-separated blood samples was developed using gray scale analysis of a point-and-shoot digital photograph of separated blood in a micochannel. Protein quantitation was subsequently performed in a high surface area-to-volume ratio microfluidic chemiluminescent immunoassay using cell depleted serum produced by microfluidic flow-based separation of whole blood samples. All three steps were achieved in a single microchannel with separation of blood samples and hematocrit evaluation in less than 1 min, and protein quantitation in 5 min.     

二元阴离子型丙烯酰胺共聚物在二价金属离子存在下相分离行为的研究

水溶性丙烯酰胺类共聚物 ,作为粘度改性剂 ,在工业上已得到广泛应用 .特别近年来 ,它们大量应用于石油工业强化采油技术 ,引起了人们很大的重视[1] .目前 ,这类用途的聚合物 ,主要在聚丙烯酰胺结构中 ,引入阴离子组分和不断增高产物分子量的方法 ,以提高聚合物溶液粘度和增粘效果 ,然而 ,在二价金属离子 (如Ｃａ2 + ,Ｍｇ2 + 等 )存在下 ,羧酸阴离子型丙烯酰胺类共聚物很容易络合发生沉淀 ,从而失去增粘作用[2 ] .同时这类聚合物中酰胺基不稳定 ,易发生水解反应转化为羧酸基 ,并随温度升高而加剧[3 ] ,因此在温度较高的应用条件下 ,二价金属…     

The Use of a 3-Factor Statistical Mixiure Design and a 2-Factor Chromatograhic Response Function to the RP-HPLC Separation of Several Free α-Amino Acids

Abstract

A 3-factor statistical mixture design and a 2-factor chromatographic response function were used in the solvent optimization of the RP-HPLC separation of a mixture containing free histidine, methionine, tyrosine, phenylalanine and tryptophan. The influences of phosphoric acid, methanol and acetonitrile on the separation were compared. A solvent condition resulting in a satisfactory separation of the amino acid mixture was predicted.     

Analytical applications of a carbon nanotubes composite modified with copper microparticles as detector in flow systems

A simple microchip electrophoresis-laser-induced fluorescence device was constructed and used for separation and determination of catecholamines. On the fabricated glass chip, an extra optical fiber insertion channel, which was perpendicular and extremely close to the separation channel, was directly integrated by nothing operations more than design features on the photomask. The utilization of optical fiber to transmit the excitation light and the integration fiber channel make the fluorescence detection system simple and disposable. For electrophoresis, optimization of separation conditions was investigated for reaching high separation efficiency and sensitivity. A separation efficiency as high as 10⁶ theoretical plate numbers could be obtained for the analytes.     

Improved liquid chromatographic method for the analysis of photosynthetic pigments of higher plants

The paper presents an improved reversed-phase LC method for the separation of the pigments from green leaves. A good separation of carotenoids and of their cis- and trans-isomers was achieved, especially for the separation of trans-lutein, zeaxanthin, cis-lutein, which are usually not well separated. No perfect separation of alpha-carotene, beta-carotene and pheophytin a was possible, but conditions for a perfect coelution of pheophytin a with either beta-carotene or alpha-carotene were established. Simultaneous equations allowing the determination of pheophytin a and alpha-carotene or pheophytin a and beta-carotene are also given.     

On the use of response surface strategy to elucidate the electrophoretic migration of carbohydrates and optimize their separation

This paper focuses on the optimization with a design of experiments of a new CE method for the simultaneous separation of four carbohydrates of interest (fructose, glucose, lactose, and sucrose) and five potentially interfering carbohydrates (ribose, xylose, maltose, mannose, and galactose) with a highly alkaline separation electrolyte for subsequent applications to food, beverage, forensic, or pharmaceutical samples. First, the factors that potentially affect the carbohydrate migration were identified: NaOH concentration in the separation electrolyte, separation temperature, and separation electrolyte conductivity. A central composite design was then carried out to determine and model the effects of these three factors on normalized migration times and separation efficiency. From the model, an optimization of the separation was carried out using a desirability analysis based on resolutions between adjacent peaks and analysis time. The optimum conditions obtained were a separation electrolyte composed of 98 mM NaOH and 120 mM NaCl to adjust the conductivity at 4.29 S/m and a separation temperature fixed at 26.5°C. Finally, these conditions were experimentally confirmed and the robustness of the obtained separation was checked.