In-channel indirect amperometric detection of heavy metal ions for electrophoresis on a poly(dimethylsiloxane) microchip

In-channel indirect amperometric detection mode for microchip capillary electrophoresis with positive separation electric field is successfully applied to some heavy metal ions. The influences of separation voltage, detection potential, the concentration and pH value of running buffer on the response of the detector have been investigated. An optimized condition of 1200 V separation voltage, −0.1 V detection potential, 20 mM (pH 4.46) running buffer of 2-(N-morpholino)ethanesulfonic acid (MES) + l-histidine (l-His) was selected. The results clearly showed that Pb²⁺, Cd²⁺, and Cu²⁺ were efficiently separated within 80 s in a 3.7 cm long native separation PDMS/PDMS channel and successfully detected at a single carbon fibre electrode. The theoretical plate numbers of Pb²⁺, Cd²⁺, and Cu²⁺ were 1.2 × 10⁵, 2.5 × 10⁵, and 1.9 × 10⁵ m⁻¹, respectively. The detection limits for Pb²⁺, Cd²⁺, and Cu²⁺ were 1.3, 3.3 and 7.4 μM (S/N = 3).     

Hydrophilic biopolymer grafted on poly(dimethylsiloxane) surface for microchip electrophoresis

A novel covalent strategy was developed to modify the poly(dimethylsiloxane) (PDMS) surface. Briefly, dextran was selectively oxidized to aldehyde groups with sodium periodate and subsequently grafted onto amine-functionalized PDMS surface via Schiff base reaction. As expected, the coated PDMS surface efficiently prevented the biomolecules from adsorption. Electro-osmotic flow (EOF) was successfully suppressed compared with that on the native PDMS microchip. Moreover, the stability of EOF was greatly enhanced and the hydrophilicity of PDMS surface was also improved. To apply thus-coated microchip, the separation of peptides, protein and neurotransmitters was investigated in detail. For comparison, these analytes were also measured on the native PDMS microchips. The results demonstrated that these analytes were efficiently separated and detected on the coated PDMS microchips. Furthermore, the relative standard deviations of their migration times for run-to-run, day-to-day, and chip-to-chip reproducibilities were in the range of 0.6-2.7%. In addition, the coated PDMS microchips showed good stability within 1 month.     

Separation of proteins on surface-modified poly(dimethylsiloxane) microfluidic devices

Separation and detection of proteins have been realized on nonionic surfactant-modified poly(dimethylsiloxane) (PDMS) microfabricated devices with end-column amperometric detection. The hydrophobic PDMS channels are turned into hydrophilic ones after being modified with Brij35 and facilitate the separation of proteins. The coating can remarkably reduce the adsorption of large protein molecules and is stable in the range of pH 6-12. The detection of proteins in such channels needs less rinsing time and thus efficiency is raised. Even large molecules of proteins can also be detected with better reproducibility and enhanced plate numbers. The relative standard deviation (RSD) of the migration time for glucose oxidase (GOD) is 2.2% (n = 19). Separation of GOD and myoglobin has been developed in modified channels. Predominant operational variables, such as the coating conditions, the concentration of surfactant and buffer, are studied in detail.     

The use of poly(dimethylsiloxane) surface modification with gold nanoparticles for the microchip electrophoresis

Poly(dimethylsiloxane) (PDMS) microfluidic channels modified by citrate-stabilized gold nanoparticles after coating a layer of linear polyethylenimine (LPEI) were successfully used to separate dopamine and epinephrine, which were difficult to be separated from baseline in native and hybrid PDMS microchannels. In-channel amperometric detection with a single carbon fibre cylindrical electrode was employed. Experimental parameters of separation and detection processes were optimized in detail. The analytes were well separated within 100 s in a 3.7 cm long separation channel at a separation voltage of +800 V using a 30 mM phosphate buffer solution (PBS, pH 7.0). Linear responses of them were obtained both from 25 to 600 μM with detection limits of 2 μM for dopamine and 5 μM for epinephrine, respectively. The modified PDMS channels have a long-term stability and an excellent reproducibility within 2 weeks.     

Ultraviolet sealing and poly(dimethylacrylamide) modification for poly(dimethylsiloxane)/glass microchips

Simple sealing methods for poly(dimethylsiloxane) (PDMS)/glass-based capillary electrophoresis (CE) microchips by UV irradiation are described. Further, we examined the possibility to modify the inner surface of separation channels, using polymethylacrylamide (PDMA) as a dynamic coating reagent. The surface properties of native PDMS, UV-irradiated PDMS, and PDMA-coated PDMS were systematically studied by atomic force microscopy (AFM), infrared absorption by attenuated total reflection infrared (ATR-IR) spectroscopy, and contact angle measurement. We found that PDMA forms a stable coating on PDMS and glass surfaces, eliminating the nonhomogeneous electroosmotic flow (EOF) in channels on PDMS/glass microchips, and improving the hydrophilicity of PDMS surfaces. Mixtures of flavin mononucleotide (FMN), flavin adenine dinucleotide (FAD), and fluorescein were separated in 35 s using PDMA-coated PDMS/glass microchips. A high efficiency of theoretical plates with at least 1365 (105 000 N/m) and a good reproducibility with relative standard deviations (RSD) below 4% in five successive separations were achieved.     

Multilayer poly(vinyl alcohol)-adsorbed coating on poly(dimethylsiloxane) microfluidic chips for biopolymer separation

A poly(dimethylsiloxane) (PDMS) microfluidic chip surface was modified by multilayer-adsorbed and heat-immobilized poly(vinyl alcohol) (PVA) after oxygen plasma treatment. The reflection absorption infrared spectrum (RAIRS) showed that 88% hydrolyzed PVA adsorbed more strongly than 100% hydrolyzed one on the oxygen plasma-pretreated PDMS surface, and they all had little adsorption on original PDMS surface. Repeating the coating procedure three times was found to produce the most robust and effective coating. PVA coating converted the original PDMS surface from a hydrophobic one into a hydrophilic surface, and suppressed electroosmotic flow (EOF) in the range of pH 3-11. More than 1,000,000 plates/m and baseline resolution were obtained for separation of fluorescently labeled basic proteins (lysozyme, ribonuclease B). Fluorescently labeled acidic proteins (bovine serum albumin, beta-lactoglobulin) and fragments of dsDNA phiX174 RF/HaeIII were also separated satisfactorily in the three-layer 88% PVA-coated PDMS microchip. Good separation of basic proteins was obtained for about 70 consecutive runs.     

Capillary zone electrophoresis of amino acids on a hybrid poly(dimethylsiloxane)-glass chip

Poly(dimethylsiloxane) (PDMS)-PDMS and hybrid PDMS-glass devices have been characterized and compared in terms of current-voltage linearity, contact angle, electroosmotic velocity, electroosmotic mobility, and electrokinetic potential in dependence on the surface treatment. The hybrid PDMS-glass microfluidic devices have further been tested as on-chip capillary electrophoresis systems for the separation of fluorescently labeled amino acids. It has been demonstrated that different methods of surface pretreatment of the PDMS-glass devices result in significantly different separation performance, with plate numbers varying from 650 to 57 000 in dependence on the surface state and the nature of the amino acids. Electrophoretic separations of amino acids have been achieved within tens of seconds with detection limits of less than 2 microM (approximately 2 x 10(-16) to 2.5 x 10(-16) mol quantities at injection volumes of 110-120 pL). The detected amounts of fluorescein isothiocyante (FITC)-amino acids are at least ten times lower, since the amino acid:FITC ratio is 10:1 mol. The results demonstrate the perspective of such hybrid PDMS-glass microfluidic systems and the methods to modify their surfaces for on-chip separation methods for biomolecules.     

Simplified current decoupler for microchip capillary electrophoresis with electrochemical and pulsed amperometric detection

There is a need to develop broadly applicable, highly sensitive detection methods for microchip CE that do not require analyte derivatization. LIF is highly sensitive but typically requires analyte derivatization. Electrochemistry provides an alternative method for direct analyte detection; however, in its most common form, direct current (DC) amperometry, it is limited to a small number of easily oxidizable or reducible analytes. Pulsed amperometric detection (PAD) is an alternative waveform that can increase the number of electrochemically detectable analytes. Increasing sensitivity for electrochemical detection (EC) and PAD requires the isolation of detection current (nA) from the separation current (muA) in a process generally referred to as current decoupling. Here, we present the development of a simple integrated decoupler to improve sensitivity and its coupling with PAD. A Pd microwire is used as the cathode for decoupling and a second Au or Pt wire is used as the working electrode for either EC or PAD. The electrode system is easy to make, requiring no clean-room facilities or specialized metallization systems. Sensitive detection of a wide range of analytes is shown to be possible using this system. Using this system we were able to achieve detection limits as low as 5 nM for dopamine, 74 nM for glutathione, and 100 nM for glucose.     

Poly(dimethylsiloxane) microchip capillary electrophoresis with electrochemical detection for rapid measurement of acetaminophen and its hydrolysate

Poly(dimethylsiloxane) microchip capillary electrophoresis with amperometric detection has been used for rapid separation and determination of acetaminophen and its hydrolysate, i.e. p-aminophenol. A Pt ultramicroelectrode with a diameter of 10 m positioned at the outlet of the separation channel was used as a working electrode for amperometric detection. Factors influencing separation and detection were investigated and optimized. Results show that acetaminophen and p-aminophenol can be well separated within 35 s with RSD<1% for migration time and <7% for detection current for both analytes. Detection limits for both analytes are estimated to be 5.0 mol L^–1 (approximately 0.1 fmol) at S/N=3. This method has been successfully applied to the detection of traces of p-aminophenol in paracetamol tablets.     

Comparison of surfactants for dynamic surface modification of poly(dimethylsiloxane) microchips

In the present report, the use of negatively charged surfactants as modifiers of the background electrolyte is reported using poly(dimethylsiloxane) (PDMS) microchips. In particular, the use of anionic surfactants, such as sodium dodecyl sulfate, phosphatidic acid, and deoxycholate, was studied. When surfactants were present in the run buffer, an increase in the electroosmotic flow (EOF) was observed. Two additional effects were also observed: (i) stabilization of the run-to-run EOF, (ii) an improvement in the electrochemical response for several biomolecules. In order to characterize the analysis conditions, the effects of different surfactant, electrolyte, and pH were studied. EOF measurements were performed using either the current monitoring method or by detection of a neutral molecule. The first adsorption/desorption kinetics studies are also reported for different surfactants onto PDMS. The separation of biologically important analytes (glucose, penicillin, phenol, and homovanillic acid) was improved decreasing the analysis time from 200 to 125 s. However, no significant changes in the number of theoretical plates were observed.     

Poly (acrylic acid) microchannel modification for the enhanced resolution of catecholamines microchip electrophoresis with electrochemical detection

A new modification of glass electrophoresis microchips based on poly (acrylic) acid immobilization has been performed. It is based on the reaction of PAA with an amine functionalized surface, obtained through the bifunctional reagent 3-aminopropyl triethoxysilane. Parameters affecting all the three steps involved: surface activation, silanization and polymer immobilization were optimized employing soda-lime glass plates. Characterization by SEM and XPS was carried out. Application of the modified microchips to the separation of a model system: dopamine (D), epinephrine (E) and norepinephrine (NE), that on the other hand are of high clinical relevance was performed employing amperometric detection. Modification is necessary for obtaining partial resolution of all the three analytes in a microchip with an effective separation length of 30 mm. Situation changes from no resolution (Rs) at all (only one peak was achieved for the mixture) to a partial resolution (Rs D–NE and Rs NE–E are 0.25 and 0.24 respectively). Microchips with 60 mm of separation channel were also modified, implying this procedure a resolution enhancement (Rs of 0.49 and 0.28 for D–NE and NE–E respectively), even when methanol is employed as organic modifier (Rs values of 0.70 (D–NE) and 0.66 (NE–E) for a 3% MeOH).     

Fabrication of a novel poly(dimethylsiloxane) microchips with two sharpened stretching tips

An integrated poly(dimethylsiloxane) (PDMS) microchip with two sharpened stretching has been presented. The sample was directly introduced into the separation channel through the stretching inlet tip without complicated power switching supplies and without injection cross-channel. Operations of running buffer refreshing or channel cleaning also becomes simple by vacuumed in one end and placed another tip into solution vial. The fabrication method can be easily applied in most analytical laboratories at low cost in the absence of soft lithography and plasma bonding equipments. Characteristics of the chips were tested and it can be used to separate fluorescence labeled molecules.     

Polycation coating poly(dimethylsiloxane) capillary electrophoresis microchip for rapid separation of ascorbic acid and uric acid

A novel method for rapid separation and determination of ascorbic acid and uric acid has been developed with a polycation-modified poly(dimethylsiloxane) (PDMS) microchip under a negative-separation electric field. Just by flushing the microchip with aqueous solutions of the polycations, poly(allylamine) hydrochloride, poly(diallyldimethylammonium chloride) or chitosan could be stably coated on the PDMS microchannel surface, which resulted in a reversed electroosmotic flow and thus the rapid and efficient separation of the two substrates. Factors influencing the separation, including polycation category, buffer solution, detection potential and separation voltage, were investigated and optimized. The cheapness, rapid analysis speed and the successful analysis of human urine make this microsystem attractive for application in clinics. Figure The electropherograms of 100 μ/mL AA and UA in (1) PAH, (2) PDDA, (3) Chitosan modified PDMS microchannels and native PDMS microchip (4).     

New preconditioning strategy for the determination of inorganic anions with capillary zone electrophoresis using indirect UV detection

It is widely accepted that preconditioning procedures are indispensable in capillary electrophoresis in order to achieve reproducibility of migration times and peak areas. Several preconditioning strategies have been employed for electrophoretic determinations of inorganic anions using indirect UV detection including simple flushing with buffer or alkaline or acid pre-rinsing followed by flushing with electrolyte. We investigated the influence of various preconditioning strategies on the reproducibility of migration times and peak areas of inorganic anions. The electrolyte systems for indirect UV detection were based on pyromellitic acid and chromic acid respectively as UV absorbing probes and hexamethonium hydroxide as electroosmatic flow modifier. Preconditioning agents under investigation were electrolyte buffer, NaOH, HCl and the free acids of the UV absorbing probes. Investigations showed that reproducibility of migration times and peak areas can be significantly improved by acid pre-rinsing using the corresponding acid of the UV absorbing probes compared to preconditioning by flushing the capillary with buffer. In contrast to acid pre-rinsing using hydrochloric acid no interfering signals within the migration time window of inorganic anions under investigation can be observed. The optimized preconditioning procedure yields relative standard deviations of migration times less than 0.25% (n=10). Relative standard deviations of corrected peak areas were below 5% applying acid preconditioning using pyromellitic acid.     

Modified Hadamard transform microchip electrophoresis

Sensitivity is a crucial point in the development applications for medicine or environmental samples in which the analytes are present in the nanomolar range. Besides further technical development of detection systems, the multiplex sample injection technique can be applied for enhancing the signal-to-noise ratio. Hadamard transform is easily applied to microchip electrophoresis due to the fact that sample injection is generally achieved through cross, double-tee, or tee injector structures. This paper reports the first demonstration of a modified Hadamard transform electrophoresis on a microchip by using an amperometric detector. Contrary to the previous Hadamard applications, the resolution (number of points per unit of time) of electropherograms obtained is independent of the number of injections.     

Application of surface plasmon resonance spectroscopy for adsorption studies of different types of components on poly(dimethylsiloxane)

Surface plasmon resonance (SPR) spectroscopy is utilized to study in real-time and, by label-free means, the reversible and quasi-irreversible adsorption of small ionic or neutral molecules, pharmaceuticals, and proteins on poly(dimethylsiloxane) (PDMS) surfaces. The SPR sensor is covered with 0.2% (w/v) PDMS in octane. During the timescale of a typical lab-on-a-chip analysis or an electrophoretic separation, it was found that small neutral components containing a hydrophobic part do not adsorb or absorb onto PDMS, while larger, water-soluble polymer-like materials like proteins generally irreversibly adsorb to PDMS. The technique can be used to monitor the kinetics of adsorption and desorption of the molecules. For the non-specific adsorption of teicoplanin to PDMS, a Langmuir-like adsorption isotherm was obtained (K_d = 32 ± 2 μmol L⁻¹).     

An end-channel amperometric detector for microchip capillary electrophoresis

An end-channel amperometric detector with a guide tube for working electrode was designed and integrated on a home-made glass microchip. The guide tube was directly patterned and fabricated at the end of the detection reservoir, which made the fixation and alignment of working electrode relatively easy. The fabrication was carried out in a two-step etching process. A 30 μm carbon fiber microdisk electrode and Pt cathode were also integrated onto the amperometric detector. The characteristics and primary performance of the home-made microchip capillary electrophoresis (MCCE) were investigated with neurotransmitters. The baseline separation of dopamine (DA), catechol (CA) and epinephrine (EP) was achieved within 80 s. Separation parameters such as injection time, buffer components, pH of the buffer were studied. Relative standard deviations of not more than 6.0% were obtained for both peak currents and migration times. Under the selected separation conditions, the response for DA was linear from 5 to 200 μM and from 20 to 800 μM for CA. The limits of detection of DA and CA were 0.51 and 2.9 μM, respectively (S/N=3).     

Electroosmotic flow-switchable poly(dimethylsiloxane) microfluidic channel modified with cysteine based on gold nanoparticles

An electroosmotic flow (EOF)-switchable poly(dimethylsiloxane) (PDMS) microfluidic channel modified with cysteine has been developed. The native PDMS channel was coated with poly(diallyldimethylammonium chloride) (PDDA), and then gold nanoparticles by layer-by-layer technique was assembled on PDDA to immobilize cysteine. The assembly was followed by infrared spectroscopy/attenuated total reflection method, contact angle, EOF measurements and electrophoretic separation methods. EOF of this channel can be reversibly switched by varying the pH of running buffer. At low pH, the surface of channels is positively charged, EOF is from cathode to anode. At high pH, the surface is negatively charged, EOF is from anode to cathode. At pH 5.0, near the isoelectric point of the chemisorbed cysteine, the surfaces of channels show neutral. When pH is above 6.0 or below 4.0, the magnitude of EOF varies in a narrow range. And the modified channel surface displayed high reproducibility and good stability, a good reversibility of cathodic-anodic EOF transition under the different pH conditions was observed. Separation of dopamine and epinephrine as well as arginine and histidine were performed on the modified chip.     

Determination of phenol in landfill leachate by using microchip capillary electrophoresis with end-channel amperometric detection

Phenol, 2,4-dichlorophenol (2,4-DCP), and 2,4,6-trichlorophenol (2,4,6-TCP) were baseline separated by using a homemade microchip CE with an end-channel amperometric detector where a 50 microm Pt microdisk working electrode (WE) and a Pt cathode were integrated onto the microchip itself. Separation parameters such as injection time and voltage, pH of the buffer, online pretreatment condition for WE, reproducibility, and detection potential were investigated. Under the selected separation conditions, the linear ranges for phenol, 2,4-DCP, and 2,4,6-TCP were 2-200, 4-400, and 4-400 microM, respectively. The LODs were 0.4, 0.5, and 0.7 microM for phenol, 2,4-DCP, and 2,4,6-TCP, respectively (S/N = 3). The standard addition method was successfully applied to the analysis of landfill leachate samples and the concentration of phenol in the landfill leachate samples was measured to be 0.32 and 0.21 mM, respectively. The recoveries were in the range of 85-103% and corresponding RSDs were less than 5.5%.     

PDMS夹心式电泳微芯片的研究

电泳微芯片由于具有自动化程度高、试剂消耗少和分析速度快等优点,目前已经成为微全分析系统研究的热点.