Quantitative Profiling

Enrichment of viable bacteria in a micro-volume by free-flow electrophoresis

Authors: 
S. Podszun, P. Vulto, H. Heinz, S. Hakenberg, C. Hermann, T. Hankemeier, G.A. Urban

Macro- to micro-volume concentration of viable bacteria is performed in a microfluidic chip. The enrichment principle is based on free flow electrophoresis and is demonstrated for Gram positive bacteria. Bacteria from a suspension flow are trapped on a gel interface that separates the trapping location from integrated actuation electrodes in order to enable non-destructive trapping. The microfluidic chip contains integrated electrolytic gas expulsion structures and phaseguides for gel and liquid handling.

DOI: 
10.1039/c1lc20575g

Limits of miniaturization: assessing ITP performance in sub-micron and nanochannels

Authors: 
K.G.H. Janssen, J.J. Li, H.T. Hoang, P. Vulto, R. van den Berg, H.S. Overkleeft, J.C.T. Eijkel, N.R. Tas, H.J. van der Linden, T. Hankemeier

The feasibility of isotachophoresis in channels of sub micrometer and nanometer dimension is investigated. A sample injection volume of 0.4 pL is focused and separated in a 330 nm deep channel. The sample consists of a biomatrix containing the fluorescently-labeled amino acids glutamate and phenylalanine, 20 attomoles of each. Isotachophoretic focusing is successfully demonstrated in a 50 nm deep channel. Separation of the two amino acids in the 50 nm deep channel however, could not be performed as the maximum applicable voltage was insufficient.

DOI: 
10.1039/c2lc21011h

An efficient hydrophilic interaction liquid chromatography separation of 7 phospholipid classes based on a diol column

Authors: 
C. Zhu, A. Dane, G. Spijksma, M. Wang, J. van der Greef, G.A. Luo, T. Hankemeier, R.J. Vreeken

A hydrophilic interaction liquid chromatography (HILIC) - ion trap mass spectrometry method was developed for separation of a wide range of phospholipids. A diol column which is often used with normal phase chromatography was adapted to separate different phospholipid classes in HILIC mode using a mobile phase system consisting of acetonitrile, water, ammonium formate and formic acid.

DOI: 
10.1016/j.chroma.2011.11.034

Nano-slit electrospray emitters fabricated by a micro- to nanofluidic via technology

Authors: 
M. Dijkstra, J.W. Berenschot, M.J. de Boer, H.J. van der Linden, T. Hankemeier, T.S.J. Lammerink, R.J. Wiegerink, M. Elwenspoek, N.R. Tas

This article presents nano-slit electrospray emitters fabricated by a micro- to nanofluidic via technology.

DOI: 
10.1007/s10404-012-0937-3

On-line large-volume electroextraction coupled to liquid chromatography-mass spectrometry to improve detection of peptides

Authors: 
P.W. Lindenburg, F.W.A. Tempels, U.R. Tjaden, J. van der Greef, T. Hankemeier

Electroextraction (EE) takes place in a two-phase liquid-liquid system, consisting of an aqueous and an organic phase, where an applied electric field causes ions to be extracted from one phase into the other, to be concentrated close after the liquid-liquid interface. The extraction takes place in a wide-bore capillary that is connected to a 2-way 10-port switching valve, which serves to couple capillary EE (cEE) with LC-MS. In this set-up, volumes as high as 100 μL can be extracted, which is a ten times larger volume than has been reported, earlier.

DOI: 
10.1016/j.chroma.2012.06.016

Feasibility of electroextraction as versatile sample preconcentration for fast and sensitive analysis of urine metabolites, demonstrated on acylcarnitines

Authors: 
P.W. Lindenburg, U.R. Tjaden, J. van der Greef, T. Hankemeier

In this work, we demonstrate the applicability of electroextraction (EE) to urine metabolites. To investigate which urine metabolite classes are susceptible to EE, off-line EE experiments were carried out with a prototype device, in which urine metabolites were electroextracted from ethyl acetate into water. The obtained extracts were examined with direct infusion MS and the results demonstrated that several compound classes could be extracted, amongst which amino acids and acylcarnitines.

DOI: 
10.1002/elps.201200276

Tunable ionic mobility filter for depletion zone isotachophoresis

Authors: 
J. Quist, P. Vulto, H.J. van der Linden, T. Hankemeier

We present a novel concept of filtering based on depletion zone isotachophoresis (dzITP). In the micro/nanofluidic filter, compounds are separated according to isotachophoretic principles and simultaneously released selectively along a nanochannel-induced depletion zone. Thus, a tunable low-pass ionic mobility filter is realized. We demonstrate quantitative control of the release of fluorescent compounds through the filter using current and voltage actuation. Two modes of operation are presented.

DOI: 
10.1021/ac301612n

Instrument and process independent binning and baseline correction methods for liquid chromatography-high resolution-mass spectrometry deconvolution

Authors: 
S. Krishnan, J.T. Vogels, L. Coulier, R.C. Bas, M.M.W.B. Hendriks, T. Hankemeier, U. Thissen

Setting appropriate bin sizes to aggregate hyphenated high-resolution mass spectrometry data, belonging to similar mass over charge (m/z) channels, is vital to metabolite quantification and further identification. In a high-resolution mass spectrometer when mass accuracy (ppm) varies as a function of molecular mass, which usually is the case while reading m/z from low to high values, it becomes a challenge to determine suitable bin sizes satisfying all m/z ranges.

DOI: 
10.1007/s11306-011-0291-6

Discovery of early-stage biomarkers for diabetic kidney disease using ms-based metabolomics (FinnDiane study)

Authors: 
F.M. van der Kloet, F.W. Tempels, N. Ismail, R. van der Heijden, P.T. Kasper, M. Rojas-Cherto, R. van Doorn, G. Spijksma, M. Koek, J. van der Greef, V.P. Mäkinen, C. Forsblom, H. Holthöfer, P.H. Groop, T. Reijmers, T. Hankemeier

Diabetic kidney disease (DKD) is a devastating complication that affects an estimated third of patients with type 1 diabetes mellitus (DM). There is no cure once the disease is diagnosed, but early treatment at a sub-clinical stage can prevent or at least halt the progression. DKD is clinically diagnosed as abnormally high urinary albumin excretion rate (AER). We hypothesize that subtle changes in the urine metabolome precede the clinically significant rise in AER. To test this, 52 type 1 diabetic patients were recruited by the FinnDiane study that had normal AER (normoalbuminuric).

DOI: 
10.1007/s11306-011-0291-6

Quantitative profiling

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