Determining the antioxidant properties of various beverages using staircase voltammetry.

Antioxidants are molecules that neutralize reactive oxygen species in the human body, reportedly reducing the risk of cancer and cardiovascular diseases. With multiple dietary products being advertised by their assumed high antioxidant concentration, the need for a proper way of analyzing antioxidant containing beverages becomes apparent. In this research, the antioxidant nature of teas, wines and (superfood) juices is investigated using staircase voltammetry (SV). A new parameter is proposed and evaluated to characterize the antioxidant nature, including its antioxidant capacity and activity: the Antioxidant Index (AI). AI showed green tea to have the best antioxidant nature of teas and red wine to be a better antioxidant than white wine. Superfoods did not show better antioxidant behavior than non-superfoods. AI proved to be a promising way of investigating the antioxidant nature of beverages.

Microfluidics and catalyst particles.

In this review article, we discuss the latest advances and future perspectives of microfluidics for micro/nanoscale catalyst particle synthesis and analysis. In the first section, we present an overview of the different methods to synthesize catalysts making use of microfluidics and in the second section, we critically review catalyst particle characterization using microfluidics. The strengths and challenges of these approaches are highlighted with various showcases selected from the recent literature. In the third section, we give our opinion on the future perspectives of the combination of catalytic nanostructures and microfluidics. We anticipate that in the synthesis and analysis of individual catalyst particles, generation of higher throughput and better understanding of transport inside individual porous catalyst particles are some of the most important benefits of microfluidics for catalyst research.

Solid contact potassium selective electrodes for biomedical applications - a review.

Ion-selective electrodes (ISE) are used in several biomedical applications, including laboratory sensing of potassium concentration in blood and urine samples. For on-site determination of potassium concentration and usage in other applications such as determination of extracellular potassium concentration, miniaturization of the sensors is required. To that extent, solid contacts have proven to be an adequate substitute of liquid contacts as inner layer for ion-to-electron transduction, allowing industrial production of miniaturized ISEs. This review paper covers relevant developments of solid-state ISEs in the past decade, critically compares current potassium ISEs and discusses future prospects for biomedical applications. Performances of three main types of solid contact materials in potassium sensing are compared, namely polypyrrole, polythiophenes and conducting nanomaterials. With these new materials, numerous improvements in stability, selectivity and time response of solid-state ISEs have been made. Current developments are new operational methods of sensing, flexible miniaturized sensors and multi-electrode designs able to measure electrolyte concentrations in one-drop blood samples or transmembrane ionic flows.

Towards microfluidic sperm refinement: impedance-based analysis and sorting of sperm cells.

The use of high quality semen for artificial insemination in the livestock industry is essential for successful outcome. Insemination using semen with a high number of sperm cells containing morphological defects has a negative impact on fertilization outcome. Therefore, semen with a high number of these abnormal cells is discarded in order to maintain high fertilization potential, resulting in the loss of a large number of morphologically normal sperm cells (up to 70-80% of original sample). A commonly occurring morphological sperm anomaly is the cytoplasmic droplet on the sperm flagella. Currently, no techniques are available to extract morphologically normal sperm cells from rejected samples. Therefore, we aim to develop a microfluidic setup which is able to detect and sort morphologically normal sperm cells label-free and non-invasively. In a proof-of-concept experiment, differential impedance measurements were used to detect the presence of cytoplasmic droplets on sperm flagella, which was quantified by calculating the area under the curve (AUC) of the corresponding impedance peaks. A receiver operating characteristic curve of this electrical analysis method showed the good predictive power of this analysis method (AUC value of 0.85). Furthermore, we developed a label-free cell sorting system using LabVIEW, which is capable of sorting sperm cells based on impedance. In a proof-of-concept experiment, sperm cells and 3 µm beads were sorted label-free and non-invasively using impedance detection and dielectrophoresis sorting. These experiments present our first attempt to perform sperm refinement using microfluidic technology.

Microfluidic single sperm entrapment and analysis.

Selection of healthy spermatozoa is of crucial importance for the success rates of assisted reproduction technologies (ART) such as in vitro fertilization and intra-cytoplasmic sperm injection. Although sperm selection for ART procedures is predominantly based on sperm motility, successful fertilization is not predicted by good motility alone. For example, sperm characteristics such as the acrosome state and DNA integrity have shown significant impact on ART outcome. Although fertilization can be achieved with a single spermatozoon of high quality, current quality assessments are population-based and do not allow investigation of multiple sperm characteristics on a single spermatozoon simultaneously. In order to study sperm cells on the single cell level, we designed and characterized a PDMS microfluidic platform that allows single sperm entrapment. After spatially confining individual sperm cells within microfluidic cell traps, the cell viability, chromosomal content and acrosome state were studied. This platform is suitable for the analysis of individual sperm cells, which could be exploited for (non-invasive) sperm analysis and selection by impedance or Raman spectroscopy.

A simple method to fabricate electrochemical sensor systems with predictable high-redox cycling amplification.

In this paper an easy to fabricate SU8/glass-based microfluidic sensor is described with two closely spaced parallel electrodes for highly selective measurements using the redox cycling effect. By varying the length of the microfluidic entrance channel, a diffusion barrier is created for non-cycling species effectively increasing selectivity for redox cycling species. Using this sensor, a redox cycling amplification of ∼6500× is measured using the ferrocyanide redox couple. Moreover, a simple, but accurate analytical expression is derived that predicts the amplification factor based on the sensor geometry.

Electrochemistry-on-chip for on-line conversions in drug metabolism studies.

We have designed an integrated 3-electrode electrochemical cell on-chip with high analyte conversion rates for use in drug metabolism studies. The electrochemical cell contains platinum working and counter electrodes and an iridium oxide pseudo-reference electrode. The pseudo-reference electrode has a pH sensitivity of -52 mV/s, and thus will provide a constant potential in solutions with known and constant pH. The average drift of the iridium oxide electrode is below 5 mV for a typical 15 min conversion experiment. We have been able to mimic the oxidative drug metabolism reactions catalysed by enzymes of the cytochrome P-450 family, normally occurring in the human body. With the chip, the different reaction products of both rat liver cell microsome and human liver cell microsome incubations have been observed.

A microfluidic chip for electrochemical conversions in drug metabolism studies.

We have designed a microfluidic microreactor chip for electrochemical conversion of analytes, containing a palladium reference electrode and platinum working and counter electrodes. The counter electrode is placed in a separate side-channel on chip to prevent unwanted side-products appearing in the measured spectrograms. Using this chip, cyclic voltammograms are measured in volumes of 9.6 nL. Furthermore the conversion efficiency of ferricyanide is characterized using UV/vis-spectroscopy. We have obtained an on-line conversion of 97% using a flow rate of 1 microL/min. We have used the microreactor chip to study the electrochemical metabolism pathway of amodiaquine using electrochemistry (EC)-liquid chromatography (LC)-mass spectrometry (MS). We have compared our results with measurements obtained with commercially available electrochemical flow-through cells. Using our chip it was possible to obtain similar results. Therefore, we have fabricated an electrochemical cell on-chip which is used successfully in EC-UV/vis and EC-LC-MS experiments.

Redox cycling with facing interdigitated array electrodes as a method for selective detection of redox species.

A pair of interdigitated ultramicroelectrodes (UMEs) is used to electrochemically detect a weak reductor (dopamine) in the presence of a stronger one (K(4)[Fe(CN)(6)]). In the mixture of both reductors, one of the two interdigitated electrodes (the generator electrode) is used to oxidize both species at 700 mV vs. Ag/AgCl, followed by subsequent (selective) reduction of the oxidized dopamine at 400 mV. A regenerated dopamine molecule can thus be oxidized several times (redox cycling) and enable selective detection even in the presence of the stronger reductor. In order to obtain high redox cycling efficiency, we designed and realized platinum electrodes with widths of 2 and 4 microm and spacing of 2 microm, which gave redox cycling efficiencies of 9 and 4 respectively. Using this electrode design, a dopamine/K(4)[Fe(CN)(6)] selectivity of 2 could be obtained.

A miniaturized carbon dioxide gas sensor based on sensing of pH-sensitive hydrogel swelling with a pressure sensor.

A measurement concept has been realized for the detection of carbon dioxide, where the CO(2) induced pressure generation by an enclosed pH-sensitive hydrogel is measured with a micro pressure sensor. The application of the sensor is the quantification of the partial pressure of CO(2) (Pco(2)) in the stomach as diagnosis for gastrointestinal ischemia. The principle is put to the proof by examining the sensor response to changes in Pco(2). Furthermore, the response time, temperature-sensitivity and resolution are determined. The sensor responds well to changes in Pco(2) with a maximum pressure generation of 0.29 x 10(5) Pa at 20 kPa CO(2). The 90% response time varies between 1.5 and 4.5 minutes at 37( composite function)C. The sensor shows a linear temperature-sensitivity which can easily be compensated for, and enables detection of Pco(2) changes as small as 0.5 kPa CO(2).

Study of chemically induced pressure generation of hydrogels under isochoric conditions using a microfabricated device.

A method is proposed to study the behavior of stimulus-sensitive hydrogels under isochoric conditions. Freedom of swell movement of such a hydrogel was restricted in all directions by enclosing the hydrogel between a micropressure sensor and a porous cover. Water and external stimuli can be applied to the hydrogel through the pores of the cover to provoke swelling, which results in pressure generation measured by the pressure sensor. The method was put to the proof by examining the response of a pH-sensitive hydrogel to changes in pH, ionic strength, and buffer concentrations of the surrounding solution. Both equilibrium and dynamic pressure generation were observed. The results show that higher pressures are obtained by incorporating more ionizable groups into the hydrogel network or by lowering the ionic strength of the external solution. Furthermore it was proven that pressures reach equilibrium faster when less titratable groups are incorporated or at the presence of higher buffer concentrations in the surrounding solution. By using microfabrication techniques the dimensions of the hydrogel could be kept small with the advantage that responses are fast. A DMAEMA-co-HEMA hydrogel with 2.5% protonable groups and a thickness of 15 microm generated a Delta pressure of 0.67 x 10(5) Pa in 12 min when a pH step from 9 to 6 was applied. The presented method is a simple and fast manner to characterize the static and dynamic stimulus-dependent behavior of hydrogels.

Highly sensitive glucose sensor based on work function changes measured by an EMOSFET.

In this paper, glucose is potentiometrically measured by using a specific field effect transistor, the EMOSFET. In this device, glucose oxidase is immobilized within a bovine serum albumin matrix, using glutaraldehyde. This layer is deposited on the top of an electroactive Os-polyvinylpyridine layer containing horseradish peroxidase, which is used as the gate material of the FET. The basic principle of the sensor is to measure the glucose concentration by means of measuring the change in the work function of the electroactive gate due to its redox reaction with the H2O2, generated by the reaction between glucose and glucose oxidase. The change in the work function can be detected as a change in the threshold voltage of the FET. Moreover, a measuring mode called "constant current potentiometry" has been applied to improve the sensitivity of the sensor. The sensitivity of the sensor working in this mode is found to be much higher than the Nernstian value. The experimental results show that the detection limit of the sensor can be tuned depending on the value of the applied current and the glucose oxidase concentration in the gate.

A flow-through amperometric sensor based on dialysis tubing and free enzyme reactors.

A generic flow-through amperometric microenzyme sensor is described, which is based on semi-permeable dialysis tubing carrying the sample to be analyzed. This tubing (300 microm OD) is led through a small cavity, containing the working and reference electrode. By filling this cavity with a few microl of an appropriate enzyme solution, an amperometric enzyme sensor results. As the dialysis tubing is impermeable for large molecular species such as enzymes, this approach does not require any immobilization chemistry, and as a consequence the enzyme is present in its natural free form. Based on this principle, amperometric sensors for lactate, glucose, and glutamate were formed by filling cavities, precision machined in Perspex, with buffered solutions containing respectively, lactate-, glucose-, and glutamate-oxidase. All sensors showed a large linear range (0-35 mM for glucose, 0-3 mM for lactate, and 0-5 mM for glutamate) covering the complete physiological range. The lower detection limit was in the order of 15-50 microM. Applicability in flow injection analysis systems is demonstrated.

An integrated micromachined electrochemical pump and dosing system.

In this paper a micromachined electrochemically driven pump capable of dosing precise nanoliter amounts of liquid is presented. The pump consists of a micromachined channel structure realized in silicon by reactive ion etching. On top of this structure a Pyrex((R)) cover piece with noble metal electrodes was bonded. The fluid to be dispensed is stored in a meander shaped reservoir which is part of the channel structure. This meander starts in an electrolyte solution containing reservoir, on top of which two noble metal electrodes are positioned. By the electrochemical production of gas bubbles by electrolysis of water at these electrodes, liquid can be driven out of the meander. The measured volume displacements were in close agreement with theory. Pump rates as low as a few nl/s could accurately be controlled via the actuation current through the electrodes. By applying current pulses rather than a continuous current, preset amounts of fluid in the nanoliter range could be dosed successfully. Because the resulting device consists of simple channel structures and metal electrodes it can easily be integrated in miniaturized chemical analysis systems to dose reagents or calibration solutions.

Constructing a Proton Titration Curve from Ion-Step Measurements, Applied to a Membrane with Adsorbed Protein

A new measuring method is described for obtaining a proton titration curve. The curve is obtained from a microporous composite membrane, consisting of polystyrene beads in an agarose matrix, with lysozyme molecules adsorbed to the bead surface. The membrane is incorporated into a sensor system by deposition on a silicon chip with a pH-sensitive ion-sensitive field effect transistor (ISFET) located in the middle of a Ag/AgCl electrode. The actual measurement is performed by creating a stepwise change in the salt concentration of the bathing electrolyte (the ion step) and measuring the ISFET potential versus the Ag/AgCl electrode. This potential shows a transient change in the ion step, which indicates a transient pH change in the membrane. This procedure is repeated at a series of pH values. Equations are presented to calculate the proton titration curve of the membrane from the amplitude and duration of the measured transients. Measurements show qualitative agreement between the curves obtained and equilibrium titration experiments on the same system.