Sander Koster

Electron capture and collisionally activated dissociation mass spectrometry of doubly charged hyperbranched polyesteramides

Electron capture dissociation (ECD) of doubly protonated hyperbranched polyesteramide oligomers (1100–1900 Da) was examined and compared with the structural information obtained by low energy collisionally activated dissociation (CAD). Both the ester and amide bonds of the protonated species were cleaved easily upon ECD with the formation of odd electron (OE·+) or even electron (EE+) fragment ions. Several mechanistic schemes are proposed that describe the complex ECD fragmentation behavior of the multiply charged oligomers. In contrast to studies of biomolecules, the present results indicate that consecutive cleavages induced by intramolecular H-shifts are significant for ECD and of less importance for low energy CAD. The capture of an electron by the ionized species results in fragmentation associated with a redistribution of the excess internal energy over the products and the subsequent bond cleavage. Low energy, multiple collision CAD is found to be a more selective dissociation method than ECD in view of the observation that only amide bonds are cleaved for most of the hyperbranched polymers examined with CAD in this study. ECD appears not to provide complementary structural information compared to CAD in the study of hyperbranched polymers, even though a significantly more complex ECD fragmentation behavior is observed. ECD is shown to be of use for the structural characterization of large oligomers that may not dissociate upon low energy CAD. This is a direct result of the fact that ECD produces ionized hyperbranched oligomers with a relatively high internal energy.

Charge-based particle separation in microfluidic devices using combined hydrodynamic and electrokinetic effects

A new microfluidic approach for charge-based particle separation using combined hydrodynamic and electrokinetic effects is presented. A recirculating flow pattern is employed, generated through application of bi-directional flow in a narrow glass microchannel incorporating diverging or converging segments at both ends. The bi-directional flow in turn is a result of opposing pressure-driven flow and electro-osmotic flow in the device. Trapping and preconcentration of charged particles is observed in the recirculating flow, under conditions where the average net velocity of the particles themselves approaches zero. This phenomenon is termed flow-induced electrokinetic trapping (FIET). Importantly, the electrophoretic mobility (zeta potential) of the particles determines the flow conditions required for trapping. In this paper, we exploit FIET for the first time to perform particle separations. Using a non-uniform channel, one type of particle can be trapped according to its zeta-potential, while particles with higher or lower zeta-potentials are flushed away with the pressure-driven or electro-osmotic components, respectively, of the flow. This was demonstrated using simple mixtures of two polystyrene bead types having approximately the same size (3 microm) but different zeta potentials (differences were in the order of 25 to 40 mV). To gain more insight into the separation mechanism, particle separations in straight, 3 cm-long microchannels with uniform cross-section were also studied under conditions of bi-directional flow without trapping. A thorough theoretical analysis confirmed that trapping occurs when electrokinetic and pressure-driven particle velocities are equal and opposite throughout the diverging segment. This makes it possible to predict the pressure and electric field conditions required to separate particles having defined zeta potentials.

Endgroup determination of synthetic polymers by electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry

Electrospray ionization (ESI) was performed on a Fourier transform ion cyclotron resonance mass spectrometer for the endgroup and monomer mass determination of three poly(oxyalkylene)s in the mass range of 400–8000 Da. A combined use of the multiple charge states observed with ESI, leads to a threefold increase in accuracy of the endgroup and monomer determination. The improvement is attributed to the increased number of datapoints used for the regression procedure, yielding more accurate results. Endgroup masses are determined with a mass error better than 5 and 75 millimass units for the molecular weight range of 400–4200 and 6200–8000 Da, respectively. A mass error of better than 1 millimass unit was observed for all monomer mass determinations. With ESI, endgroup and monomer masses have been determined for poly(ethylene glycol) oligomers with a mass higher than 8000 Da. This is almost two times higher than observed with matrix-assisted laser desorption/ ionization on the same instrument.

An Enzymatic Microreactor Based on Chaotic Micromixing for Enhanced Amperometric Detection in a Continuous Glucose Monitoring Application

The development of continuous glucose monitoring systems is a major trend in diabetes-related research. Small, easy-to-wear systems which are robust enough to function over many days without maintenance are the goal. We present a new sensing system for continuous glucose monitoring based on a microreactor incorporating chaotic mixing channels. Two different types of chaotic mixing channels with arrays of either slanted or herringbone grooves were fabricated in poly(dimethylsiloxane) (PDMS) and compared to channels containing no grooves. Mixing in channels with slanted grooves was characterized using a fluorescence method as a function of distance and at different flow rates, and compared to the mixing behavior observed in channels with no grooves. For electrochemical detection, a thin-film Pt electrode was positioned at the end of the fluidic channel as an on-chip detector of the reaction product, H(2)O(2). Glucose determination was performed by rapidly mixing glucose and glucose oxidase (GOx) in solution at a flow rate of 0.5 microL/min and 1.5 microL/min, respectively. A 150 U/mL GOx solution was selected as the optimum concentration of enzyme. In order to investigate the dependence of device response on flow rate, experiments with a premixed solution of glucose and GOx were compared to experiments in which glucose and GOx were reacted on-chip. Calibration curves for glucose (0-20 mM, in the clinical range of interest) were obtained in channels with and without grooves, using amperometric detection and a 150 U/mL GOx solution for in-chip reaction.

Structural characterization of hyperbranched polyesteramides: MSn and the origin of species

Four hyperbranched synthetic polyesteramides were synthesized by the polycondensation of the trifunctional diisopropanolamine (D) and difunctional anhydrides (X) of succinic acid, glutaric acid, 1,2-cyclohexane dicarboxylic acid, and phthalic acid. The polymers were analyzed with electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry. The most intense oligomer series observed was XnDn+1 containing diisopropanolamine end groups as expected from the polycondensation conditions. A series of oligomers XnDn+1H2O is observed as well, which can have its origin in the polymerization process or alternatively could result from in-source fragmentation of XnDn+1. Breakdown diagrams of the protonated parent ions X3D4 and additional MSn (n = 1, 2, 3) measurements gave insight in the fragmentation behavior of the polymers. Three main fragmentation pathways have been observed for all polymers of which the loss of H2O to oxazolonium ions has the lowest onset energy followed by the rearrangement of the amide and ester bonds also leading to oxazolonium ions. The loss of a second H2O to allylic or morpholine end groups has highest onset energy. MS3 experiments demonstrated that the presence of a series of oligomers XnDn+1H2O can be attributed to the polymerization process. Most probably an allylic end group has formed from one of the alcohol end groups. The formation of allylic end groups partly terminates the polymerization reaction and results in a change of the composition of the molecular weight distribution and decrease of the number average molecular weight.

Microfabricated platforms for the study of neuronal and cellular networks

In this contribution we present the development of three microfabricated devices for the study of neuronal and cellular networks. Together, these devices form an attractive toolbox, which is useful to stimulate and record signals of both electrical and chemical nature. One approach consist of microelectrode arrays for the study of neuronal networks, and allow for the electrical stimulation of individual cells in the network, while the other electrodes of the array record the electrical activity of the remaining cells of the network. We also present the use of micropipettes that can measure the extra- and intracellular concentrations of ions in cells cultures. A third approach exploits the laminar flows in a microfluidic device, to deliver minute amounts of drug to some cells in a cellular network. These three illustrations show that microfabricated platforms are appealing analytical tools in the context of cell biology.

Behaviour of Human Umbilical Vein Endothelial Cells (HUVEC) Cultivated in Microfluidic channels

Our long-term goal is to develop advanced tools for cell studies and analysis based on microfluidic systems. In this paper, we report on endothelial cell cultivation in microchannels and 96-well tissue plates, and compare cell phenotype and cellular status in the two environments This was done under both pro-inflammatory conditions (cell stimulation in the presence of the cytokine, Tumour Necrosis Factor, TNFalpha) and normal (medium only) conditions. In addition, we considered the behaviour of cells in a pro-inflammatory environment in the presence of an anti-inflammatory drug