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1. Preface 2. Programme at a glance 3. Detailed programme 4. Abstracts plenary sessions 5. Abstracts parallel programme 6. Poster list 7. Map `de Werelt'

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On behalf of the NWO study group in Analytical Chemistry, I would like to welcome you to a special edition of the "Lunteren Meeting". This year we have joined hands with the Section Analytical Chemistry (SAC) of the Royal Dutch Chemical Society (KNCV) to organize the Total Analytical Challenge 2010 (TAC2010). During this event we will celebrate the start of the first Top Institute of Comprehensive Analytical Sciences and Technology (COAST) in the Netherlands. I wish all of you a stimulating and fruitful meeting and hope that the feeling of being one joint analytical science community will continue to grow throughout the years to come.

Rainer Bischoff Chairman of the NWO study group

After the success of last year I am very pleased to welcome you at the second edition of The Analytical Challenge, TAC2010. This year, the Section Analytical Chemistry (SAC) of KNCV joined forces with the Analytical Chemistry group of NWO-CW in order to again create an event where analytical chemists from academia and industry and manufacturers of analytical equipment can meet and learn from the latest developments in our exciting work field. Next to this, TAC2010 will be the kick-off of COAST, the fantastic new top institute on analytical sciences. On behalf of the SAC board I wish you all (a) very pleasant and stimulating day(s) and I hope that this event will contribute to a further strengthening of Analytical Chemistry in the Netherlands. Harry Philipsen Chairman SAC


Programme at a glance

Monday 1-nov-10

9:00 9:10 9:20 9:30 9:40 9:50 10:00 10:10 10:20 10:30 10:40 10:50 11:00 11:10 11:20 11:30 11:40 11:50 12:00 12:10 12:20 12:30 12:40 12:50 13:00 13:10 13:20 13:30 13:40 13:50 14:00

Coffee Prepare Posters Opening Paul Haddad Chair: Thomas Hankemeier

Azie Amerika Amerika

Andreas Nasioudis (AkzoNobel) Coffee Prepare Posters Clare Harvey (Weckhuysen) Eslam Nouri-Nigjeh (Bischoff) Elena Uliyanchenko (Schoenmakers)



Amerika Amerika Amerika



Chair: Bert Kip

Martin Giera (VU)


14:10 14:20 14:30 14:40 14:50 15:00 15:10 15:20 15:30 15:40 15:50 16:00 16:10 16:20 16:30 16:40 16:50 17:00 17:10 17:20 17:30 17:40 17:50 18:00 18:10 18:20 18:30 18:40 18:50 19:00 19:10 19:20 19:30 19:40 19:50 20:00 20:10 20:20 20:30 20:40 20:50 21:00

Lygia Azevedo Marques (Irth) Marcel van Batenburg (Smilde) Robert-Jan Raterink (Hankemeier) NWO talent session & Coffee POSTERS Clemens Heilmann (de Koster) Chair: Rainer Bischoff Bregje de Kort (de Jong) Ivonne Lammers (Gooijer) Maarten Altelaar (Heck) Gabriel Vivó Truyols (UvA)

Amerika Amerika Amerika

Azie /Afrika

Amerika Amerika Amerika Amerika Amerika

Dinner & projectleaders meeting

Restaurant /Afrika

New members (coordinated by Heiko vd Linden)


Maurice Aalders End



Programme at a glance

Tuesday 2-nov-10

9:00 9:10 9:20 9:30 9:40 9:50 10:00 10:10 10:20 10:30 10:40 10:50 11:00 11:10 11:20 11:30 11:40 11:50 12:00 12:10 12:20 12:30 12:40 12:50 13:00 13:10 13:20 13:30 13:40 13:50 14:00 14:10 14:20 14:30 14:40 14:50 15:00 15:10 15:20 15:30 15:40 15:50 16:00 16:10 16:20 16:30 16:40 16:50 17:00 17:10 17:20 17:30 17:40 17:50 18:00 18:10 18:20 18:30 18:40 18:50 19:00 19:10 19:20 19:30 19:40 19:50 20:00 20:10 20:20 20:30 20:40 20:50 21:00 Afrika Inez Finoulst (Verhaert) Jose Castro-Perez (Hankem.) A. Smolinska (Buydens) Opening SAC Cees Gooijer Amerika (also visible in Afrika) Koffie SAC (Exhibition) Afrika

Coffee (Exhibition) Prepare posters Forensics - Afrika P. Zoon WSM - zaal 4+5 NMR DG - Amerika M. Heringa E. Golovina DSP- Vide 1+2 J. Purmova

Restaurant/ Azie

B. Venhuis

Afrika/zaal 4+5/ Amerika/Vide 1+2

E. Kaal

A.J. Oosthoek

J. de Heer

Lunch POSTERS (Exhibition)


G. Counotte

J. Wieling

J. van Duynhoven

G. Saunders Afrika/zaal 4+5/ Amerika/Vide 1+2

A. Nuijs


E. Talnishnikh

B. Staals

C. Brasseur

Serena di Palma

T. Didenko

P. Cools

Coffee (Exhibition) R. Van Rijn


L. Switzar

A.J. Bakermans

A.J. de Graaf Afrika/zaal 4+5/ Amerika/Vide 1+2

C. Otto

R. Haselberg

E. van Eck

A. Chojnacka

Jacques Joosten Launch TI-COAST Drinks (Exhibition)




Detailed programme

NWO Plenary Sessions Monday 1 November 2010 Time Title 09.30 ­ 10.00 10.00 ­ 10.10 10.10 ­ 10.50 Coffee / Prepare Posters Speaker Room Azië



Separation of inorganic ions by ion chromatography and capillary electrophoresis. Which is best? Discrimination of Polymers by Using Their Characteristic Collision Voltage in Tandem Mass Spectrometry Coffee / Prepare Posters Integrated In-Situ AFM-Raman Study of SERS Effects on Supported Silver Catalyst Nanoparticles Electrochemistry in the mimicry of oxidative drug metabolism by Cytochrome P450s Ultra-performance liquid chromatography for polymer seperations Lunch / POSTERS

Paul R. Haddad, University of Tasmania, Australia Andreas Nasioudis, AkzoNobel Research


10.50 ­ 11.10 11.10 ­ 11.20 11.20 ­ 11.50 11.50 ­ 12.10 12.10 ­ 12.30 12.30 ­ 13.50


Azië Clare Harvey, Utrecht University Eslam Nouri-Nigjeh, University of Groningen Elena Uliyanchenko, University of Amsterdam Amerika



Restaurant/ Azië


NWO Plenary Sessions Monday 1 November 2010 14.00 14.30 14.30 ­ 14.50 14.50 ­ 15.10 15.10 ­ 15.30 15.30 ­ 16.10 Novel Derivatization strategies for the LC-ESI-MS measurement of relevant biomarkers and acidic metabolites Stability-Indicating Analysis of the Acetylcholinesterase Inhibitors (-) Huperzine A, Tacrine and Galantamine New figures of merit for comprehensive functional genomics data: the metabolomics case Chip-based heaterless nano-APCIMS NWO talent session & coffee / POSTERS Martin Giera, VU University Amsterdam L.A. Marques, VU University of Amsterdam Marcel van Batenburg, University of Amsterdam Robert-Jan Raterink, Utrecht University Amerika



Amerika Azië/Afrika

16.10 ­ 16.30 16.30 ­ 16.50

To be announced Probing protein conformation and purity by capillary electrophoresis with wavelength-resolved native fluorescence detection Enantioselective phosphorescence detection coupled to capillary electrophoresis Electron transfer dissociation; enabling improved peptide dissociation To be announced Dinner & projectleaders meeting New members To be announced

Clemens Heilmann, University of Amsterdam Bregje de Kort, Utrecht University

Amerika Amerika

16.50 ­ 17.10 17.10 ­ 17.30 17.30 17.50 17.50 19.30 19.30 20.10 20.10 20.50 ­ ­ ­ ­

Ivonne Lammers, VU University of Amsterdam Maarten Altelaar, Utrecht University Gabriel Vivo Truyols, University of Amsterdam


Amerika Amerika Restaurant/ Afrika Amerika Amerika

Heiko vd Linden Maurice Aalders


NWO Plenary Sessions Tuesday 2 November 09.00 ­ Mass spectrometry as alternative 09.20 detection system for cytokines secreted by human t-cells 09.20 ­ The Importance of Early 09.40 Atherosclerosis Biomarker Screening in Target Identification and Validation 09.40 ­ 1H-NMR spectroscopy and pattern 10.00 recognition methods for metabolomics investigation of preclinical model of Multiple Sclerosis Opening KNCV programme 10.1010.50 Thrusting Back Frontiers of Analytical Biomolecular Spectroscopy Cees Gooijer Amerika Inez Finoulst, Delft University Jose-Perez Castro, Leiden University A. Smolinska, Radboud University Nijmegen Afrika Afrika


Parallel Programme KNCV


The crucial role of Analytical Science and Technology for Innovations in the Dutch Chemical Sector

Jacques Joosten



Parallel Programme TAC 2010 2 november 2010

Forensics - Parallel

Subject/title Speaker Chair: A. van Asten 11.15-11.45 Scanning Electron Microscopy in Forensic Science -Visualising the Invisible 11.45-12.15 12.15-13.30 Fraudulent medicines: A decade of fake Viagra B. Venhuis, RIVM Afrika Restaurant/Azië P. Zoon, NFI Afrika Room

Lunch & Posters / Exhibition Chair: B. de Rooij


Analytical challenge in veterinary toxicology The analysis of wastewater as an alternative way to estimate illicit drug consumption Forensic GCxGC-TOFMS study of cadaveric volatile organic compounds (VOCs) released by buried decaying pig carcasses

G. Counotte, Gezondheid V. Dieren A.L.N. van Nuijs, TNO Def





C. Brasseur, Un. Liege



Coffee Break / Exhibition Chair: H-J van Lent


15.30-16.00 16.00-16.30

Radiology beyond the grave, a forensic perspective Molecular detection in- and outside Biological Cells

R. Van Rijn, AMC C. Otto, Universiteit Twente

Afrika Afrika

Chair: E.Kellenbach 16.30-17.15 17.15-17.25 17:25-18:00 Plenary: Jacques Joosten Launch TI-COAST (Members only) Drinks / Exhibition Restaurant/ Azië Amerika


Werkgroep Scheidingsmethoden ­ parallel

Title/Subject Speaker Sample prep / tox analyse in Minne Heringa, water with LC-MS KWR Nieuwegein Biomarker and profiling Erwin Kaal, strategies for the diagnosis of University of Tuberculosis using GC and Amsterdam GC×GC-TOF-MS Lunch, posters / exhibition "Dried Blood Spot sampling in Jaap Wieling, early clinical development QPS Groningen studies for pharmacokinetics, (formerly: Xendo) pharmacogenomics and safety assessments" HILIC Adema, Leiden University ZIC-HILIC as a 1st dimension Serena di Palma, in a two-dimensional liquid Utrecht University chromatography configuration provides high resolution separation and increased sensitivity in proteome analysis Coffee / exhibition Optimization of protein Linda Switzar, digestion for the VU Amsterdam characterization of drug-protein adducts Characterization of Rob Hasselberg, biopharmaceuticals by capillary Utrecht University electrophoresis ­ mass spectrometry Plenary: Jacques Joosten Launch TI-COAST (Members only) Drinks/exhibition Room Zaal 4 + 5 Zaal 4 + 5

11.15 ­ 11.45 11.45 ­ 12.15

12.15 ­ 13.30 13.30 ­ 14.00

Restaurant/Azië Zaal 4 + 5

14.00 ­ 14.30 14.30 ­ 15.00

Zaal 4 + 5 Zaal 4 + 5

15.00 - 15.30 15.30 ­ 16.00

Restaurant/Azië Zaal 4 + 5

16.00 ­ 16.30

Zaal 4 + 5

16.30 ­ 17.15 17.15 - 17.25 17.25 - 18.00

Amerika Restaurant/Azië


NMR­ parallel

11.15 ­ 11.25 11.25 ­ 11.45 Subject / title Opening NMR-DG symposium ESR and NMR are complementary approaches in the study of the physical-chemical predictors of seed longevity Reaction kinetics in microfluidic NMR Speaker Room Amerika Amerika

Elena Golovina, Wageningen University

11.45 ­ 12.05

12.05 ­ 13.30 13.30 ­ 14.00

14.00 ­ 14.20

14.20 ­ 14.40

14.40 ­ 15.00

15.00 ­ 15.30 15.30 ­ 16.00

16.00 ­ 16.20

16.30 ­ 17.15 17.15 - 17.25 17.25 - 18.00

Anna Jo Oosthoek - de Vries, Radboud University Nijmegen Lunch, posters / exhibition Identification of John van Duynhoven, polyphenol metabolites in Unilever biofluids by LC-SPEcryoNMR-MS Proton NMR relaxometry Elena Talnishnikh, of leaves under severe Wageningen University water stress in low magnetic fields. 3D DOSY-TROSY to Tanja Didenko, determine the Utrecht University translational diffusion coefficient of large protein complexes In vivo assessment of A.J. Bakermans, triglyceride content in the Eindhoven University mouse heart Coffee / exhibition Studying molecular Ernst van Eck, ordering by 13C solid Radboud University state NMR: arrangement Nijmegen of amphiphiles, motions in molecules, and packing of polymers Proton NMR relaxometry Elena Talnishnikh of leaves under severe Ageningen University water stress in low magnetic fields Plenary: Jacques Joosten Launch TI-COAST (Members only) Drinks/exhibition


Restaurant/Azië Amerika




Restaurant/Azië Amerika


Amerika Restaurant/Azië


DSP ­ Parallel

Subject/title Speaker Chair: E.Mes 11.15-11.45 Protein Analysis by Size Exclusion Chromatography Automated (multivariate) data analysis for SEC Jindra Purmová, AkzoNobel Chemicals Jos de Heer, Sabic Innovative Plastics Vide 1 + 2 Room

11.45-12.15 12.15-13.30

Vide 1 + 2 Restaurant/Azië

Lunch & Posters / Exhibition Chair: E.Gelade


Characterization of maltodextrin ­ acrylic hybrid copolymers Using an Evaporative Light scattering Detector (ELSD) for quantification with LC, a blessing or a curse? Industrial applications of SEC hyphenated with ESITOF-MS

Greg Saunders, Agilent Technologies Bastiaan Staals,

Vide 1 + 2


Vide 1 + 2


Paul Cools, DSM

Vide 1 + 2


Coffee Break / Exhibition Chair: H.Brouwer



Challenges in molecular weight determination of PEG-pNIPAm block copolymers Study of solubility and dissolution of polymers

A.J. de Graaf

Vide 1 + 2


Aleksandra Chojnacka

Vide 1 + 2

Chair: E.Kellenbach 16.30-17.15 17.15-17.25 17:25-18:00 Plenary: Jacques Joosten Launch TI-COAST (Members only) Drinks / Exhibition Restaurant/ Azië Amerika


Abstracts plenary sessions 1-2 November 2010

Monday 1 November

P1: Separation of inorganic ions by ion chromatography and capillary electrophoresis. Which is best? Paul R. Haddad Discrimination of Polymers by Using Their Characteristic Collision Voltage in Tandem Mass Spectrometry A. Nasioudis; A. Memboeuf; R. M. A. Heeren; D. F. Smith; K. Vékey; L. Drahos; O. F. van den Brink Integrated In-Situ AFM-Raman Study of SERS Effects on Supported Silver Catalyst Nanoparticles Clare E. Harvey, Evelien M. van Schrojenstein Lantman, Arjan J. G. Mank, and Bert M. Weckhuysen Electrochemistry in the mimicry of oxidative drug metabolism by Cytochrome P450s Eslam Nouri-Nigjeh, Hjalmar P. Permentier, Rainer Bischoff, Andries P. Bruins ULTRA-PERFORMANCE LIQUID CHROMATOGRAPHY FOR POLYMER SEPARATIONS Elena Uliyanchenko Novel Derivatization strategies for the LC-ESI-MS measurement of relevant biomarkers and acidic metabolites Giera M., Kloos D. P., Kretschmer A., Wijtmans M., Lingeman H., Niessen W.M.A., Irth H. Stability-Indicating Analysis of the Acetylcholinesterase Inhibitors (-) Huperzine A, Tacrine and Galantamine Marques L.A., Maada I., Giera M., Kool J., de Kanter F.J.J., Lingeman H., Niessen W.M.A. and Irth H. New figures of merit for comprehensive functional genomics data: the metabolomics case M.F van Batenburg, Leon Coulier, Fred van Eeuwijk, Age K. Smilde, and Johan A. Westerhuis CHIP-BASED HEATERLESS NANO-APCI-MS Robert-Jan Raterink




P5 P6




P10: Quantitative strategies to map the Candida albicans cell wall proteome Clemens Heilmann

P11: Probing protein conformation and purity by capillary electrophoresis with wavelengthresolved native fluorescence detection Bregje J. de Kort, Gerhardus J. de Jong and Govert W. Somsen P12 Enantioselective phosphorescence detection coupled to capillary electrophoresis I. Lammers, J.B. Buijs, F. Ariese, C. Gooijer

P13: Electron transfer dissociation; enabling improved peptide dissociation A.F. Maarten Altelaar P14: Gabriel Vivo Truyols P15: Maurice Aalders P16: MASS SPECTROMETRY AS ALTERNATIVE DETECTION SYSTEM FOR CYTOKINES SECRETED BY HUMAN T-CELLS Inez Finoulst, Paul Vink, Mervin Pieterse, Martijn Pinkse, Ebo Bos and Peter Verhaert P17: The Importance of Early Atherosclerosis Biomarker Screening in Target Identification and Validation Jose-Castro-Perez P18:


H-NMR spectroscopy and pattern recognition methods for metabolomics investigation of pre-clinical model of Mutiple Sclerosis. A. Smolinska, L. Blanchet, K. Ampt, A. Attali, T.Luider, A. van Gool, S.S. Wijmenga and L.M.C Buydens

Tuesday 2 November

P19: THRUSTING BACK FRONTIERS OF ANALYTICAL BIOMOLECULAR SPECTROSCOPY Cees Gooijer P20: The crucial role of Analytical Science and Technology for Innovations in the Dutch Chemical Sector Dr. Jacques Joosten (Director Technology at DSM, Managing Director DSM Dutch Polymer Institute and vice chairman of the National Chemistry Board (`Regiegroep Chemie')

P1: Separation of inorganic ions by ion chromatography and capillary electrophoresis. Which is best? Paul R. Haddad Australian Centre for Research on Separation Science, University of Tasmania, Private Bag 75, Hobart 7001, Australia. [email protected] Ion chromatography (IC) and capillary electrophoresis (CE) are used routinely for the determination of inorganic anions and cations. Both techniques have their inherent strengths and weaknesses. In this presentation, IC and CE are compared in terms of their stage of development, separation efficiency, separation selectivity, analytical performance parameters, method development procedures, applications, and adoption into regulatory methods. Throughout this comparison, the techniques will be evaluated in the context of their use for the determination of inorganic anions and cations in the identification of inorganic improvised explosives. These are formed from commonly available ingredients, such as fertilizers, and have been used extensively in acts of terrorism. Current counter-terrorism measures require rapid and reliable detection of these improvised explosives both prior to detonation (preblast) and after detonation (post-blast). The primary aim of pre-blast identification is to detect the explosive in situations such as airport screening, while the primary aim of postblast identification is to determine the identity of the particular explosive used in order to assist in apprehension of the perpetrators. Both situations require reliable and rapid means of analysis using methods which can be operated by relatively unskilled personnel and in field-based locations. A survey of improvised explosives has been undertaken and a suite of 15 candidate inorganic anions and 13 candidate cations has been established for post-blast fingerprinting. The separations of each group of ions has been developed using IC and CE, with an emphasis on the use of instrumentation which is field-deployable or portable. To this end, miniaturised columns and robust detection methods, such as indirect spectrophotometric detection using light-emitting diode detectors have been employed. These separations have been applied to traces of the explosives and also to post-blast explosive residues and have been evaluated for their capacity to unequivocally identify the particular explosive used. P2: Discrimination of Polymers by Using Their Characteristic Collision Voltage in Tandem Mass Spectrometry A. Nasioudis1; A. Memboeuf2,4; R. M. A. Heeren3; D. F. Smith3; K. Vékey2; L. Drahos2; O. F. van den Brink1

1 2

AkzoNobel Research, Development & Innovation, Deventer, The Netherlands; Chemical Research Center, Hungarian Academy of Sciences, Budapest, Hungary; 3 FOM Institute for Atomic and Molecular Physics, Amsterdam, The Netherlands; 4 Joseph Fourier University, Department of Molecular Chemistry, Grenoble, France The characteristic collision voltage to obtain 50% fragmentation (CCV) was used as a tool to discriminate between different classes of polymers. The CCV value of different polymers was determined in a quadrupole ion trap mass spectrometer. A good linear correlation (0.980<R2<0.999) between the CCV values and precursor ion mass was found for all polymers studied (Figure 1). The position of the various linear trend lines varied among the various polymers, which allowed their grouping based on the respective CCV values (Figure 1). The collision energy necessary to drive fragmentation was decreasing in the order of: polyethers > polymethacrylates > polyesters > polysaccharides. This suggests that polysaccharides fragment most easily (low CCVs), while polyethers require the highest collision energy among the polymers studied. The effect of endgroup on the CCV was also studied, showing minor influence in most cases. In addition the applicability of CCV as discriminator was studied for a poly(lactic acid)-block-poly(tetramethylene glycol)-blockpoly(lactic acid) block copolymer. Differences between the CCV values of four nominally isobaric polymers (of which two copolymers, and two homopolymers) were observed. These results demonstrate that the insertion of a "weak" link into a relatively "strong" polymer chain significantly affects the energy required for fragmentation.

Figure 1. Characteristic collision voltage (CCV) vs the precursor ion masses of the studied singly lithium cationized polymers: Polyethyleneglycol dimethylether (MePEGMe), poly(ethylene glycol) (PEG), poly(tetramethyleneglycol) (PTMEG), poly(methyl methacrylate) (PMMA), poly(butylenes adipate) diol (PBAd), cyclic poly(lactic acid) (CPLA), Maltodextrin (Maltodex), and linear poly(lactic acid) (PLA). P3: Integrated In-Situ AFM-Raman Study of SERS Effects on Supported Silver Catalyst Nanoparticles Clare E. Harvey, Evelien M. van Schrojenstein Lantman, Arjan J. G. Mank, and Bert M. Weckhuysen Debye Institute of Nanomaterials Science, Utrecht University Supported metal nanoparticles are important heterogeneous catalysts in many industrial processes. It is particularly important to have a thorough understanding of which specific surfaces have the highest catalytic activity in order to tune the shape and size of nanoparticles to achieve maximum catalytic activity. Raman spectroscopy has proven to be a powerful and versatile tool for the study of chemical reactions, particularly when utilizing the surface enhanced Raman scattering (SERS) capabilities of noble metals. The integration of atomic force microscopy (AFM) with Raman spectroscopy forms a powerful new tool for nanoscale chemical imaging of catalytic solids, allowing nano-scale morphological features to be correlated to chemical information. Here we demonstrate the potential of this integrated approach for identifying exactly which nano-particle morphologies are the most catalytically active through a study of the photo-oxidation of Rhodamine-6G over Ag nanoparticles. We show that it is possible to follow the reactants or products of a reaction under in-situ conditions, and pinpoint the active section of the catalyst on the nano-scale.

Figure 1. (A) TEM image of 30 nm Ag cubes; (B) AFM scan of Ag cubes deposited on Al2O3 surface with spin coated Rhodamine-6G; and (C) SERS spectrum of Rhodamine-6G over a cluster of Ag cubes.


Electrochemistry in the mimicry of oxidative drug metabolism by Cytochrome P450s Eslam Nouri-Nigjeh, Hjalmar P. Permentier, Rainer Bischoff, Andries P. Bruins Analytical Biochemistry and Mass Spectrometry Core Facility, Department of Pharmacy, University of Groningen, A. Deusinglaan 1, 9713 AV Groningen, The Netherlands The mimicry of oxidative drug metabolism by Cytochrome P450s (CYP) is important in the early stages of drug development. To mimic oxidative drug metabolism, in this study, we utilize electrochemically generated reactive oxygen species (ROS), as a surrogate for the reactive species involved in CYP. ROS was generated in a two-compartment electrochemical cell to separate the oxidation products from direct oxidation (anodic) and reaction by ROS (cathodic). The reaction between ROS and test drug compounds, e.g. lidocaine, was studied through electrochemical techniques, and oxidation products obtained under different conditions (potential, atmosphere), were analyzed and characterized using LC-MS(/MS). The results of this study proved that a combination of direct oxidation and reactions with electrochemically-generated ROS offers access to a wide range of drug metabolites.


ULTRA-PERFORMANCE LIQUID CHROMATOGRAPHY FOR POLYMER SEPARATIONS Elena Uliyanchenko Analytical-Chemistry Group, van 't Hoff Institute for Molecular Sciences (HIMS), Faculty of Science, University of Amsterdam, Science Park 904, 1078 XH Amsterdam, The Netherlands e-mail: [email protected] Liquid chromatography at ultra-high pressures (above the 400 bar commonly used in HPLC) has become popular for a large range of applications. It is often used in life-science, medical, food and environmental analyses. Because of the higher pressure limits, it allows the use of columns packed with sub-2-m particles. This, in turn, improves the efficiency of separations and/or reduces the analysis time. Thus, UPLC offers advantages for the analysis of various types of samples. Polymers show particular behaviour during chromatographic separations. The peak width in polymer separations is determined both by the chromatographic column and conditions and by the sample polydispersity. Previous work (1) has led to the suggestion that the separation of large molecules at very high (reduced) flow rates may be relatively favourable. However there is limited information available on the analysis of polymers using Ultra-High Pressure Liquid Chromatography. One possible obstacle may be the degradation of high-molecularweight polymers at high shear rates. This problem may become more important if high pressures are applied during separations. This complicating factor for the separation of polymers at high linear velocities has already been identified by Stegeman (2). In this work we study possibilities and limitations of UPLC system for separations of (high molecular weight) polymers. Using polystyrene standards at size-exclusion conditions we can observe some selectivity for the low- molecular-weight polymers. The selectivity for highermolecular-weight polymers in the hydrodynamic mode is lower than that in the size-exclusion range. We attempt to define chromatographic conditions and system parameters to improve the quality of separations and to prevent possible degradation of macromolecules. 1. S.-T. Popovici, W.Th. Kok and P.J. Schoenmakers, J. Chromatogr. A 1060 (2004) 237252 2. G. Stegeman, J.C. Kraak and H. Poppe, J. Chromatogr. 550 (1991) 721

P6: Novel Derivatization strategies for the LC-ESI-MS measurement of relevant biomarkers and acidic metabolites Giera M.1, Kloos D. P.1, Kretschmer A.1, Wijtmans M.2, Lingeman H.1, Niessen W.M.A.1, Irth H.1 VU University Amsterdam, Faculty of Science, BioMolecular Analysis Group, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands 2 VU University Amsterdam, Faculty of Science, Department of Pharmacochemistry, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands Carboxylic acids and aldehydes form two large groups of biologically important molecules either as biomarkers of oxidative stress originating from arachidonic acid, or as cellular metabolites, such as the intermediates of the tricarboxylic acid cycle (TCA cycle). In the analysis of both analyte classes, there are still drawbacks in terms of chromatographic behaviour, and mass spectrometric response. Therefore, we developed different derivatization strategies in order to allow the determination of these compounds by LC/MS analysis in positive mode electrospray ionization (ESI+). For the classes of acid or aldehyde containing biomarkers, we have developed new derivatization reagents based on a primary amine group, which we successfully applied to the determination of biomarkers, such as n-alkyl aldehydes and isoprostanes. The different labelling strategies and reagents will be discussed. Recently, we have started to investigate the possibilities of LC/MS based derivatization strategies in the field of targeted (cellular) metabolite analysis. Especially in connection with the rather novel field of systems biology, more sensitive and accurate measurements of predefined sets of metabolites are required. We have applied the previously developed strategy for carboxylic acids to the intermediates of the TCA cycle. A modified label with a secondary amine group was chosen to prevent side reactions and allow complete labelling even of tri-carboxylic acids such as citrate. In this lecture, we discuss the different coupling strategies for aldehydes as we well as acids. Examples are given for the measurement of biomarkers and cellular metabolites. P7: Stability-Indicating Analysis of the Acetylcholinesterase Inhibitors (-) Huperzine A, Tacrine and Galantamine Marques L.A.1, Maada I.1, Giera M.1, Kool J.1, de Kanter F.J.J.2, Lingeman H.1, Niessen W.M.A.1 and Irth H.1

1 VU University Amsterdam, Faculty of Sciences, BioMolecular Analysis Group, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands 2 VU University Amsterdam, Faculty of Sciences, Division of Organic and Inorganic Chemistry, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands 1

Stability analysis of drug substances is crucial for registration procedures. Degradation products have to be fully characterized not only in a chemical but also in a biological way. In the here presented study, we investigated the stability of the three anti- Alzheimer's drugs tacrine, (-) huperzine A and galantamine. We investigated the stability of the three aforementioned substances according to ICH guidelines and determined the kinetics of the different degradation processes. All formed degradation products were chemically and biochemically characterized, elucidating their structure and defining their inhibitory activity against the target enzyme Acetylcholinesterase (AChE). For tacrine, a huge number of degradation products were found under oxidative conditions, (-) huperzine A formed a new photoisomer (photohuperzine A) under irradiation and galantamine showed degradation under irradiation, as well as oxidative and acidic conditions.


New figures of merit for comprehensive functional genomics data: the metabolomics case M.F van Batenburg, Leon Coulier, Fred van Eeuwijk, Age K. Smilde, and Johan A. Westerhuis Swammerdam Institute for Life Sciences, University of Amsterdam Netherlands Metabolomics Centre, Leiden TNO Quality of Life, Zeist Applied Statistics, Wageningen UR Functional genomics data require figures of merit for diagnosing the quality of a measurement. In the field of metabolomics, hundreds of metabolites are measured simultaneously by analytical platforms such as GC-MS, LC-MS and NMR to obtain their concentration levels in a reliable way. A measure of analytical repeatability is the precision of a metabolite concentration. The measurement errors, used for estimating this precision come from a sample that is analyzed repeatedly in one series. However, measurement error is not constant since its value may depend on the concentration level of the metabolite. A widely used figure of merit for non-constant measurement errors is Relative Standard Deviation (RSD) but the precision predicted on the basis of this figure of merit underestimates the true value at low and high concentrations. Measurement errors may also be correlated between different metabolites. Standard multivariate analyses of concentrations of different metabolites may therefore produce erroneous results. Here we introduce new figures of merit for comprehensive data that describe the key characteristics of non-constant (hetero-scedastic) measurement error variance and concentration dependent correlation between measurement errors. They find their origin in a model for hetero-scedastic measurement error variance of each metabolite and a significance test for measurement error correlations in a metabolomics data set. The median and 90% level of the distribution of the parameter describing the hetero-scedasticity across all metabolites are used as summary statistics. The figure of merit for error correlations is a pvalue related to testing the significance of the error correlations. We used metabolomics GC-MS data as an example to illustrate our new figures of merit. We found a 90% level that is 3.5 times the median level. So 10% of the peaks have very large non-constant measurement errors. The data set has also a p-value below significance level, and indicates the presence of error correlations. Such correlations are the result of data pretreatments that in theory may remove all trends due to experimental errors but in practice fail to do so for several metabolites.


CHIP-BASED HEATERLESS NANO-APCI-MS RJ Raterink1,2, M. de Korte1, H. van der Linden1,2 and T. Hankemeier1,2

1 2

Department of Analytical Biosciences, LACDR, Leiden University, the Netherlands Netherlands Metabolomics Centre, Leiden University, the Netherlands

Electrospray Ionization-Mass Spectrometry (ESI-MS) is widely used for analyzing samples. However, ESI is burdened by low ionization efficiency, ion suppression and is restricted to polar solvents. APCI is another ambient ionization technique which has the advantage of in principle high ionization efficiency and suffers from much less from ion suppression compared to ESI. These highly-desired properties however are jeopardized by the fact that the sample is usually introduced at a very high flowrate and the solvent has to be heated to evaporate the solvent. Therefore conventional APCI cannot handle low sample volumes and thermolabile molecules. Our novel heaterless nano-APCI source overcomes all these problems. Our prototype design was fabricated in COC-4 plastic substrates using micromilling. The chip consisted of a channel structure layer and a solid capping layer. The heaterless nano-APCI source was tested successfully in front of a triple-quad MS by direct infusion of a reserpine solution. The MS showed a clear reserpine peak in the spectrum. The heaterless nano-APCI source has a number of important advantages, i.e. (i) simple onchip integration (no HV connection and no heater electrodes on chip are required), (ii) low flow rates, (iii) room temperature operation (no sample degradation and no sample clogging due to crystallization).

Our new miniaturized APCI source does not require heating of the solvent feed going into the ion source. Because no heating is required now also thermally labile compounds can be in principle ionized with the ion source using low flowrates without degradation. In this way the benefits of APCI can be fully exploited at the nanoflow range possibly surpassing nano-ESI. This new source is very promising for mass spectrometry in general but holds great potential specifically for the life sciences where MS analyses are often done on thermally labile biological compounds such as proteins, peptides and metabolites. Further research is done on optimization of different spray geometries and characterization of the source. P10: Quantitative strategies to map the Candida albicans cell wall proteome Clemens J. Heilmann, Alice G. Sorgo, Henk L. Dekker, Chris G. de Koster, Frans M. Klis and Leo J. de Koning University of Amsterdam A variety of strategies are currently available for relative and absolute quantification of proteins but their use for biological research is still in its infancy. We have developed a strategy based on 15N stable isotope labeling and high mass accuracy Fourier Transform Mass spectrometry for relative quantification. This strategy was applied to the cell wall proteome of the human opportunistic fungal pathogen Candida albicans. One of the most important features of C. albicans is its ability to switch between yeast and hyphal growth. We prepared the walls pertaining to four hyphal induction conditions and then measured them against 15N-reference walls derived from a mix of both growth forms. The walls related to each condition were mixed with the 15N-reference walls 1:1, trypsinized and the resulting peptides were analyzed on an LC-FT-ICR-MS. The "inhouse" developed CoolToolBox software identified the co-eluting 14N/15N peptide pairs according to their accurate mass tag and retention time and calculated their ratios. To validate the retention time tags and the identification, MS/MS runs of the reference walls were performed. All peptide ratios of one protein were combined and directly compared to the other conditions. In total, we were able to quantify up to 22 wall proteins. The abundance of these wall proteins can be useful to select new diagnostic markers and novel targets for vaccine development. In addition, by absolutely quantifying the 15N reference culture with 14N AQUA peptides, we were able to extrapolate the absolute abundance of three wall proteins in all conditions. In summary, by combining the two strategies of stable isotope labeling and AQUA quantification, we were able to absolutely quantify wall proteins of C. albicans. These data will be used in the ongoing Marie Curie FINSysB consortium to identify potential vaccine targets and clinical markers. P11: Probing protein conformation and purity by capillary electrophoresis with wavelength-resolved native fluorescence detection Bregje J. de Kort, Gerhardus J. de Jong and Govert W. Somsen Biomolecular Analysis, Utrecht University, the Netherlands The assessment of purity and molecular conformation (unfolding, misfolding, or aggregation) of proteins is of particular importance in the biochemical and biopharmaceutical fields. However, the determination of compositional and conformational integrity of proteins is a challenging analytical issue. Usually, various analytical techniques are required for a full protein characterisation. On-line combination of techniques for protein analysis can be an effective approach to enhance the information content that can be achieved in a single run. Capillary electrophoresis (CE) is a very attractive separation technique for purity and stability analysis of proteins, as it offers fast and efficient separations and requires only small sample volumes. Protein emission spectroscopy, based on the native fluorescence of aromatic amino acid residues, is a selective and sensitive detection technique which can provide information on protein conformation. This presentation outlines the on-line coupling of CE with wavelength-resolved fluorescence detection (WRFlu) for native protein analysis. WRFlu is accomplished using a dedicated fluorescence cell that employs wave-guiding principles to lead protein emission light to a spectrograph with CCD detector. The cell is installed in a commercial CE instrument enabling the recording of full protein spectra in a routine fashion. The new CE-WRFlu set-up allows fast separation and detection of proteins down to 10 nM, permitting detection of minor protein impurities and degradation products. Moreover, it will be shown that changes in protein conformation can be probed by the recorded emission characteristics as well as the measured electrophoretic mobility. The ability to monitor protein folding/unfolding kinetics will be demonstrated for test proteins,

such as carbonic anhydrase II and -lactoglobulin B. The presented results will indicate that CE-WRFlu is a highly promising tool for the quality assessment of intact proteins. P12: Enantioselective phosphorescence detection coupled to capillary electrophoresis I. Lammers, J.B. Buijs, F. Ariese, C. Gooijer Biomolecular Analysis and Spectroscopy, Laser Centre, Vrije Universiteit Amsterdam, the Netherlands Currently, in biomedical and pharmaceutical analysis there is much interest in enantioselectivity since the two enantiomers of a chiral compound generally have different biological activities. Though enantioselective separation techniques (in liquid chromatography and capillary electrophoresis (CE)) are well-developed and very efficient, appropriate enantioselective detection methods are still lacking. The currently available chiroptical techniques (optical rotation dispersion and circular dichroism) suffer from a limited sensitivity and are therefore difficult to couple with micro-separation techniques. Enantioselective cyclodextrin-induced room-temperature phosphorescence (CD-RTP) in the liquid state opens perspectives here. We demonstrate the coupling of this novel enantioselective detection method to the electrophoretic separation of (+)- and (-)camphorquinone (CQ). Elegantly, both the chiral separation and the enantioselective detection arise from the complexation with -CD. The difference in complexation constants of (+)- and (-)-CQ with -CD results in their electrophoretic separation in the presence of negatively charged CM--CD. Focusing on the detection part, the inclusion into -CD protects the CQ enantiomers differently against phosphorescence quenching from the environment. The resulting difference in phosphorescence lifetimes of (+)- and (-)-CQ is the basis for the on-line distinction between the enantiomers. Different phosphorescence detection modes will be discussed for the detection of CQ enantiomers after their separation in CE: quenched, direct1 and sensitized phosphorescence. The enantioselectivity and sensitivity will be compared with special attention for the influence of CM--CD used for the separation. The developed methods are used to determine the enantiomeric impurity in commercial standards of (+)- and (-)-CQ and to quantify the amount of CQ enantiomers leaching from a cured dental resin.

[1] I. Lammers, J. Buijs, G. van der Zwan, F. Ariese, C. Gooijer, Anal. Chem. 81 (2009) 6226 P13: Electron transfer dissociation; enabling improved peptide dissociation A.F. Maarten Altelaar Biomolecular Mass Spectrometry and Proteomics Group Recently, electron transfer dissociation (ETD) of peptides was introduced as an alternative peptide fragmentation method. ETD cleaves peptides at the N-C bond producing c'-and ztype ions and can be complementary to CID since it prefers larger and more basic peptides, which attain multiple charges during electrospray ionization (ESI). Interestingly, ETD leaves PTMs largely intact on the peptide backbone during fragmentation thus providing, potentially, simpler spectra in which the site of modification can be easily annotated. Here we will discuss the merits and potential drawbacks of ETD in the analysis of such `atypical' peptides and

present the use of an alternative protease (Lys-N) to produce favorable peptides for ETD sequencing. P14: Gabriel Vivo Truyols P15: Maurice Aalders P16: MASS SPECTROMETRY AS ALTERNATIVE DETECTION SYSTEM FOR CYTOKINES SECRETED BY HUMAN T-CELLS Inez Finoulst1, Paul Vink2, Mervin Pieterse1, Martijn Pinkse1, Ebo Bos2 and Peter Verhaert1

1 Dept. of Biotechnology, Analytical Biotechnology Section & the Netherlands Proteomics Centre, Delft University of Technology, Delft, The Netherlands; [email protected] 2 Department of Immune Therapeutics, Merck Research Laboratories, MSD, Oss, the Netherlands

We here report the application of mass spectrometry (MS) for detecting human cytokines in culture media. We focused on interleukin-2 (IL-2), secreted by activated T-cells, as example. The human immune system relies on the proper functioning of white blood cells or leukocytes. By using an intricate set of cytokines, different types of leukocytes regulate the defence of the body against infectious diseases and foreign 'intruders'. Of these leukocytes, T-lymphocytes or T-cells are among the best described key players in the immune system. Tlymphocytes are distinct from other lymphocytes by their T-cell receptor (TCR). Upon activation, they secrete IL-2, which stimulates long-term in vitro T-cell growth. Monitoring Tcell activation is typically done by mRNA expression and/or immunological techniques, such as ELISA or FACS. With the sensitivity of modern mass spectrometers, the direct detection of secreted cytokines via MS becomes an attractive alternative. Moreover, it potentially detects many low abundant proteins simultaneously without the need of specific antibodies. We here demonstrate this by the mass spectrometric detection of IL-2 in activated human primary T-cell supernatants. After isolation from a buffy coat, CD4+CD25- human T-cells are stimulated using beads coated with anti-CD3/anti-CD28. Two days after stimulation, the cells are checked by flow cytometry for the presence of activation markers. Next, cells are centrifuged and supernatant is filtered (0.22µm pore size) and analyzed for IL-2 by ELISA. Following, the supernatant is chromatographed by RP-HPLC on a microbore C4 column. Collected fractions are trypsin digested after reduction and alkylation. Tryptic peptides are analyzed by nanoflow LC MS/MS on an Orbitrap Velos (ThermoFisher). FACS shows an increased expression of both CD-25 and CD-69 on activated T-cells, respectively the IL-2 receptor and an early T-cell activation marker. ELISA data demonstrate that the supernatant of activated cells contains 1nM IL-2, whereas from non-activated cells IL-2 amounts in the media remains undetectable. Since serum albumin (HSA) is the most abundant protein present in the culture medium used (between 0.1-1 mg/ml), the MS/MS data were mostly dominated with tryptic peptides from HSA. In a second stage, samples were measured employing an exclusion list for the tryptic HSA ions identified in the first round. Database searches identified IL-2 exclusively in the activated primary T cells supernatant. Using this MS approach as low as 750 pg IL-2 can be detected from culture medium in the presence of 5-6 orders higher amounts of HSA, and this without the need of any depletion. P17: The Importance of Early Atherosclerosis Biomarker Screening in Target Identification and Validation Jose Castro-Perez1, David McLaren1, Stephen Previs1, Thomas P. Roddy1 Rob Vreeken2, Thomas Hankemeier2

1 2

Merck & Co., Inc. Atherosclerosis Exploratory Biomarkers, Rahway, NJ 07065, USA Analytical Biosciences & Netherlands Metabolomics Centre, Leiden, The Netherlands

Lipid analysis plays an increasingly important role in the understanding and diagnosis of cardiovascular disease. Lipid profiling in atherosclerosis drug discovery whether targeted or untargeted is key to provide an insight in the mechanism of action and to prove target engagement. This is particularly important in the early stages of target identification and validation. Furthermore, metabolic tracing of pathways and flux analyses plays a pivotal role

because it allows for early indication of lipid target efficacy and aid in the interpretation of phenotypical differences when steady state concentration measurements are insufficient. In this paper we will present a number of examples using UPLC/high resolution TOF MS which highlights how this technology may be applied to exploratory biomarkers of atherosclerosis. The examples will include; perturbation of diet, genetic knockdowns, metabolic tracers and metabolic flux of lipids. P18:

1 H-NMR spectroscopy and pattern recognition methods for metabolomics investigation of pre-clinical model of Mutiple Sclerosis. A. Smolinska1, L. Blanchet1, K. Ampt1, A. Attali2, T.Luider3, A. van Gool4, S.S. Wijmenga1 and L.M.C Buydens1

Radboud University Nijmegen, Institute for Molecules and Materials (IMM), Heyendaalseweg 135, 3525 AJ Nijmegen, The Netherlands 2 Abbott Healthcare Products B.V., Weesp, The Netherlands 3 Erasmus Medical Center, Rotterdam, The Netherlands 4 Molecular Profiling, MSD, Singapore, Singapore The composition of biofluids carries invaluable information about the biochemical status of a living organism. Cerebrospinal Fluid (CSF) is the biofluid, which is in closest interaction with the central nervous system (CNS). It is therefore the biofluid that best mirrors the biochemical status and processes in brain and central nervous system. The chemical composition of CSF may thus provide insights about metabolic pathways in the CNS. The comprehensive analysis of CSF may define the fingerprint of neurological diseases such as the Multiple Sclerosis (MScl). Our objective is to detect molecular biomarkers for MScl in CSF (and/or plasma). 1H Nuclear Magnetic Resonance (1H -NMR) spectroscopy (together with chemometric analysis) is an efficient and unbiased tool to detect and quantify metabolites in biofluids. The research results we will present here are from a pre-clinical animal model on the progression of the MScl disease; the acute Experimental Autoimmune/Allergic Encephalomyelitis (EAE) was used as an (rodent/rat) animal model for MScl. The rats were either healthy or in different stages of the disease. The 1H-NMR spectra contain several thousands of intensities coming from many metabolites and are therefore numerically complex. PLS-DA and ANOVA-PCA [1] were used to find subset of metabolites differentially profiled across the control and EAE-affected animals, and to identify relevant, disease-specific metabolites. In addition the correlation network between relevant metabolites was calculated. [1] J.R de Haan et al, Interpreation of ANOVA models for microarray data using PCA. Bioinformatics 23 (2) 2007, 184-190. P19: THRUSTING BACK FRONTIERS OF ANALYTICAL BIOMOLECULAR SPECTROSCOPY Cees Gooijer Biomolecular Analysis and Spectroscopy LaserLAB, VU, Amsterdam It is the challenge of analytical biomolecular spectroscopy to develop and implement analytical methodologies directed at the detection and structural characterization of biologically active molecules. To fully characterize such molecules, eventually not only the molecular properties as such are important, but also the (stereo selective) dynamic interaction with their immediate environment. Molecular spectroscopy methods are widely involved in this field, in particular absorbance and fluorescence spectroscopy. Unfortunately, the former one has a limited sensitivity, while the latter one ­ though providing excellent sensitivity and better selectivity than absorbance ­ is characterized by a low spectral resolution. The other mode of molecular luminescence with a high analytical potential, i.e. molecular phosphorescence is hardly invoked yet; it is generally seen as not being applicable to liquid solutions. Furthermore, Raman Spectroscopy (RS), the vibrational technique of choice for aqueous samples, is still scarcely playing a role in analytical biomolecular spectroscopy. As a technique based on inelastic scattering, it suffers from a poor sensitivity so that (too) high analyte concentrations are needed; furthermore in practice RS spectra are frequently overwhelmed by fluorescence. In this presentation we will briefly discuss recent achievements in our group (supervised by Dr. Freek Ariese and Dr. Gert van der Zwan), directed on thrusting back the frontiers of the above molecular spectroscopy methods: A laser-based alternative absorbance detection method, i.e. Cavity Ring Down Spectroscopy in the evanescence mode to directly monitor the


adsorption of protein (monolayers) to capillary walls, a main point of concern in protein capillary electrophoresis (CE). High-resolution fluorescence to study the structure of ligandreceptor complexes. Room temperature phosphorescence as a sensitive detection method for CE providing enantioselectivity based on phosphorescence lifetimes. By using Resonance Raman Spectroscopy in the deep-UV the RS sensitivity limitation is largely tackled. Furthermore we have successfully involved a combined electrochemistryResonance Raman Spectroscopy approach to obtaine detailed information about redoxprotein-ligand interactions. Last but not least we are currently developing time-gated RS, enabling the recording of RS of fluorescent proteins. Fascinating is the possibility to use it for depth profiling purposes by making use of differences in photon travelling times. P20: The crucial role of Analytical Science and Technology for Innovations in the Dutch Chemical Sector Dr. Jacques Joosten (Director Technology at DSM, Managing Director DSM Dutch Polymer Institute and vice chairman of the National Chemistry Board (`Regiegroep Chemie') Following the start of the concept of key areas by the Dutch Innovation Platform in 2005 the National Chemistry Board (NCB) has defined the ambitions and developed a series of initiatives to strengthen the position of the Dutch Chemical Industry in a global context for the key area Chemistry. In a first step, the ambitions for the Dutch chemical sector were formulated in a business plan: The National Chemistry Board aims to achieve a sustainable economic growth expressed by two objectives: - double the contribution of the Chemical sector to the Dutch GDP in 10 years. - halve the use of fossil materials (both as feed stock as well as energy) in 25 years These challenging aims will be tackled both by creating the conditions to improve innovations at the parties involved (e.g. human capital agenda, valorisation programs) and by directly stimulating innovation in public-private R&D programs. Various programs have been developed like the Sectorplan Physics and Chemistry, the Polymer Innovation Program (PIP), Dutch Separation Technology Institute (DSTI), The Institute for Sustainable Process Technology (ISPT) and ACTS etc.. In the presentation, Jacques Joosten will give an overview of the ambitions and the related programs. In this context, the NCB has recognized the importance of enabling technologies like Analytical Science and Technology. The NCB has welcomed and strongly supported the COAST (Comprehensive Analytical Science and Technology) initiative aiming at strengthening Analytical Science and Technology in the Netherlands. The availability of adequately trained employees at professional bachelor, master and PhD level and the creation of break-throughs in Analytical Science.are very important factors for innovations in the chemical arena, in the Analytical Technology sector itself and in Dutch society.

Abstracts Parallel Programme TAC 2010 2 november 2010

Forensics: No abstracts

NMR1: ESR and NMR are complementary approaches in the study of the physicalchemical predictors of seed longevity. Elena Golovina, Pieter de Waard, Magda Witek, Folkert Hoekstra and Henk Van As Lab of Biophysics and Wageningen NMR Centre, Wageningen University, Wageningen The structural and chemical stabilities of dry seeds are the basis of their longevity. Temperature-induced changes in ESR spectra from membrane and cytoplasmic spin probes provide information about the structural and chemical stabilities of membranes and cytoplasm in dry cells, respectively. Temperature-induced free radical signal in dry seeds can be used to estimate the total resistance of the matrix in dry seeds to oxidation. NMR-spectroscopy supplies information about compounds in seed extracts, which might relate to seed longevity. Solid-state NMR allows in-vivo estimation of the sensitivity of seed lipids to oxidation. Altogether, ESR and NMR approaches give a set of data, which can be used to predict seed longevity. NMR2: Reaction kinetics in microfluidic NMR A.J. Oosthoek - de Vries1, J. Bart1, P.J.M. van Bentum1, J.W.G. Janssen1, J.G.E. Gardeniers2, A.P.M. Kentgens1. [1] Radboud University Nijmegen, Institute for Molecules and Materials, Nijmegen, The Netherlands [2] Mesa+ Inst. for Nanotechnology, Mesoscale Chemical Systems, University of Twente, The Netherlands NMR spectroscopy is a versatile tool for chemical analysis and structure determination. In recent years, much interest has been shown in scaling down the conventionally used sample volume (500 l) in liquid NMR to microliter or nanoliter volumes, by using a microcoil set up. An improved design for microcoil NMR is the so called rf-stripline [1]. Using the rf-stripline a microfluidic NMR on a chip probe [2,3] has been developed. The microfluidic stripline NMR chip enables 1H-NMR measurements on 286 nl sample volumes, with high resolution and sensitivity, in a regular 600 MHz NMR spectrometer. For in-situ analysing the reaction, a Micronit microreactor chip (Y-junction type) was implemented on top of the probe.Two syringe pumps are used to control the flow of the reactants. The reaction starts in the reaction channel of the microreactor and subsequently the products flow through the stripline where the detection takes place. By changing the flow rate, the reaction time can be varied from several seconds up to 10 minutes with fast single scan detection sensitivity. As a basis for further research, we studied the reaction of benzyl alcohol with acetyl chloride in the presence of DIPEA. Considering our findings, a reaction mechanism is proposed where benzyl alcohol reacts with ketene and/or the DIPEA-acetate complex into benzyl acetate. More research is presently ongoing to solve the details of the reaction mechanism.


Figure 1: Spectra of the acetylation of benzyl alcohol in the presence of DIPEA, reaction time from 4.2 seconds to 5.6 minutes.

[1] P.J.M. van Bentum et al J. Magn. Reson., vol. 189, pp. 104-113, 2007. [2] J. Bart et al J. Am. Chem. Soc., vol. 131, pp. 5014-5015, 2009. [3] J. Bart et al J. Magn. Reson., vol. 201, pp. 175-185, 2009. NMR3: Identification of polyphenol metabolites in biofluids by LC-SPE-cryoNMR-MS Klinkenberg, M., de Roo, Niels, Alexandre, P., Mahlous, I., Janssen, H.-G., Jacobs, D., van Duynhoven, J., Unilever Discover Vlaardingen, The Netherlands Polyphenols are an important class of functional ingredients that are currently being investigated for their potential health benefits. Upon consumption, polyphenols undergo conjugation in the human host and/or bioconversion by the colonic microbiota. Many of the resulting metabolites are not know and their identification is a considerable challenge. In many cases one can take recourse to spectral databases (NMR or MS), but often one needs to embark on a de-novo molecular identification. This requires isolation and enrichment of the metabolite(s) of interest and subsequent structural elucidation by a combination of both NMR and MS. An efficient metabolite identification platform is provided by combining high-performance LC with NMR and MS. The increased automation and the incorporation of on-line solid-phase extraction (SPE) into an integrated system recently improved the detection limits. A further gain in sensitivity is provided by the use of cryogenic NMR probeheads. LC-SPE-cryoNMR-MS platforms have already been deployed for identification of metabolites from single drug compounds dosed at high levels. Nutritional polyphenol formulations are much more complex and dosages are mostly low. Hence we needed to design a strategy for identification of low-abundance metabolites against the highly complex metabolic background of biofluids. This strategy has successfully been applied to identify metabolites that are being produced upon microbial fermentation of green tea in a colonic in-vitro model. A range of low-abundance metabolites has been identified which could be used to further expand the known colon microbial degradation pathway of epicatechines in green tea. The identification approach is now being extended for identification of polyphenol metabolites in urine, and first results will be presented. NMR4: Proton NMR relaxometry of leaves under severe water stress in low magnetic fields. Elena Talnishnikh and Henk Van As Lab of Biophysics and Wageningen NMR Centre, Wageningen University, Wageningen NMR techniques to determine porosity and pore-size distribution are well-established in geology and well-logging. There porosity can be determined directly from the NMR signal amplitude of the fluid in the rock if normalized correctly. On one hand it looks very attractive to use similar approach in order to follow water status in plants and to study different compartments based on water distribution in leaves. On the other hand complex leaf structure and shrinking-elongation movements in combination with magnetic field inhomogeneities (which is often a characteristic of low field NMR equipment) make the application of portable NMR to biological system more complicated. Nowadays the research remains limited in this area, especially in the area of non-destructive and in vivo experiments with low field NMR applied to intact leaves. (to the best of our knowledge only one paper is now by now). Our first goal was to run reproducible and simple experiments with more or less standard and cheap equipment. So it can be of direct interest for a biologist. The other aim of this work was to study dehydration of various


plant species such as bean, poplar, ficus, oak, and others. In this part of the work leaves were wilted naturally in a laboratory under constant external conditions. The dehydration of leaves was followed continuously from the moment they were removed from a plant until their mass became constant. At the same time water status of detached leaves were measured with two portable unilateral NMR systems which differ in field strength and field homogeneity. Then NMR measurements were compared with weight measurements of leaves. At some point the dehydration process was subject to manipulation too. Leaves were not only air dried: some of them were exposed to osmotic stress by placing them in polyethylene glycol solution and then measured as described above. In all cases time evolution of the NMR signal amplitudes and corresponding T2 distributions showed good correlation with dehydration process in leaves. Results suggest that correlation of NMR signal with the water status of leaves depends on signal-to-noise ratio (expectably) and on the type of plants (not-obvious). NMR5: 3D DOSY-TROSY to determine the translational diffusion coefficient of large protein complexes Tanya Didenko, Rolf Boelens and Stefan Rudiger Utrecht University The translational diffusion coefficient is a sensitive parameter to probe conformational changes in proteins and protein-protein interactions. Pulsed field gradient NMR spectroscopy allows to measure the translational diffusion with high accuracy. Twodimensional heteronuclear NMR spectroscopy combined with diffusion-ordered spectroscopy (DOSY) provides improved resolution and therefore selectivity as compared to a conventional one-dimensional readout. Here we show that a combination of selective isotope labeling, 2D 1H-13C methyl-TROSY and DOSY allows to study diffusion properties of large protein complexes. We propose that a 3D DOSY-HMQC pulse sequence, that uses the TROSY effect of the HMQC sequence for 13C methyl-labelled proteins, is highly suitable for measuring the diffusion coefficient of large proteins. We used the 20 kDa cochaperone p23 as model system to test this 3D DOSY-TROSY technique under various conditions. We determined the diffusion coefficient of p23 in viscous solutions, mimicking large complexes of up to 200 kDa. We found the experimental data to be in excellent agreement with theoretical predictions. To demonstrate the use for complex formation, we applied this technique to record the formation of a complex of p23 with the molecular chaperone Hsp90, which is around 200 kDa. We anticipate that 3D DOSY-TROSY will be a useful tool to study conformational changes in large protein complexes. In addition we applied this technique to study conformational changes in 170kDa molecular chaperone Hsp90.


In vivo assessment of triglyceride content in the mouse heart A.J. Bakermans1, S.M. Houten2, T.R. Geraedts1, M. van Weeghel2, K. Nicolay1, And J.J. Prompers1


Biomedical NMR, Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven 2 Laboratory Genetic Metabolic Diseases, Academic Medical Center, Amsterdam

Introduction Accumulation of potentially toxic lipid metabolites may be a key contributor to the pathogenesis of hypertrophic cardiomyopathy - a common clinical feature of inherited mitochondrial fatty acid -oxidation (FAO) disorders. In this in vivo NMR study, the long chain acyl-coenzyme A dehydrogenase knockout (LCAD-/-) mouse model was used to investigate morphology, function and triglyceride (TG) content of the FAO-impaired heart in a non-invasive fashion. Methods Animals: Male LCAD-/- mice and C57BL/6 wild type mice underwent the MR protocol in the fed state to acquire baseline data. Two weeks later, the same animals were fasted for 24h prior to fasted state MR experiments. MR protocol: Anesthetized mice were positioned supine into a horizontal bore 9.4T MR system (Bruker BioSpin). Heart rate (>500 min-1) and respiration were used for MR gating/triggering. A birdcage coil (Ø 35mm) was used for RF transmission and signal reception.


Prospective triggered cine MR image series were acquired in 5-7 contiguous 1mm slices in the left ventricular (LV) short-axis orientation together with two- and four-chamber long axis views. Imaging parameters: FOV=30×30mm2, matrix size=192×192, TE/TR=1.8ms/7ms, flip angle=15º, NA=6. Localized 1H-MRS was performed in the diastolic interventricular septum using a respiratory gated and cardiac triggered point resolved spectroscopy (PRESS) sequence preceded by a chemical shift selective (CHESS) water suppression module. PRESS parameters: voxel size=1×2×2mm3, TE/TR=9.1ms/~2s. During respiratory gates, dummy PRESS pulses were transmitted to maintain steady state required for quantification of metabolite concentrations. Data analysis: Ejection fraction (EF) and LV myocardial mass were determined from the cine MR images using semiautomatic segmentation software (Pie Medical Imaging). MR spectra were analyzed using AMARES in jMRUI. Metabolite concentrations were quantified relative to the unsuppressed water peak amplitude measured in the same voxel. Results LV myocardial mass was higher in LCAD-/- mice than in controls (P<0.01), indicating LCAD-/- LV hypertrophy. EF decreased after fasting in LCAD-/- mice (P<0.01), accompanied by elevated levels of myocardial TG (+63%, P<0.05). In wild type mice, fasting did not affect EF, whereas TG levels dropped (-40%, P<0.05). Discussion Fasting increased myocardial TG content in LCAD-/- mice, accompanied by decreased LV EF. Combined, these findings may point towards toxic effects of lipids accumulating in the FAO-impaired myocardium. NMR7: Studying molecular ordering by 13C solid state NMR: arrangement of amphiphiles, motions in molecules, and packing of polymers Ernst RH van Eck Radboud University Nijmegen, Institute for Molecules and Materials, Nijmegen WGSM1: Extraction of water for both chemical and toxicological analysis Minne Heringa, Erik Emke, Margo van der Kooi, Ariadne Hogenboom KWR Watercycle Research Institute Until recently, the chemical quality of drinking water and its sources was mainly monitored by chemical analysis. Now, toxicological analyses are gaining ground in a complementary effect-directed approach, detecting all harmful compounds and their total mixture effect. To enable the analysis of a water sample by both a broad-screening chemical method (e.g. LC-MS) and a toxicological test, a sample preparation method suitable for both types of analyses was sought. Different extraction materials were compared in the recovery of a set of around 30 compounds, varying in hydrophobicity and functional groups. Additionally, the elution solvents were studied, as well as the best way to concentrate the extract without losing too many volatile components. An important issue rose on the preferred use of DMSO for toxicity tests, while DMSO gives problems in LC-MS analyses. Finally, a method using one broad-spectrum column has been chosen and validated in both toxicity tests and chemical analysis. This method will of course not fully extract all present compounds, but provides the best possible balance between practicality, costs and extraction efficiency. WGSM2: Biomarker and profiling strategies for the diagnosis of Tuberculosis using GC and GC×GC-TOF-MS Erwin Kaala,b, Sjaak de Koningc, Arend Kolka,d and Hans-Gerd Janssena,e


Van 't Hoff Institute for Molecular Sciences, University of Amsterdam. DSM Biotechnology, Delft. c Leco cooperation, Mönchengladbach, Germany. d KIT Biomedical Research, Royal Tropical Institute, Amsterdam. e Unilever Research and Development, Vlaardingen.


There is an increased demand for fast detection and identification of bacteria causing diseases. Of particular interest to human health is the diagnosis of mycobacteria tuberculosis (M. Tuberculosis). The present methods, mainly X-ray and microscopy,


suffer from important limitations because they are not specific and sensitive, but slow, labor intensive and/or expensive in terms of running costs. A promising approach is the use of GC hyphenated to thermochemolysis (THM). In previous work, an in-liner THM ­ GC-MS method was developed for rapid diagnosis of M. Tuberculosis in sputum. This new in-liner THM-method was based on direct liquid injections of dispersed bacteria into the GC. Drying of the sample, addition of the reagent, incubation, and derivatization were performed inside the liner of a programmable temperature vaporizer (PTV) - injector. Because of the complex sample matrix, the obtained profiles were rather complex. Fortunately, specific biomarkers could be used for rapid and reliable identification of the mycobacteria. Although the use of biomarkers results in a fast and reliable method, finding the markers is difficult and time consuming. Moreover, the markers might not be 100% specific. Therefore, using complete compound profiles instead of a limited set of biomarkers is preferable. To improve the quality of the profiles more resolution is needed. This can be obtained with two dimensional GC preferably in combination with ToF mass spectrometric detection. In the present study, we studied the potentials of THM ­ GC×GC-TOF-MS for profiling bacteria. A set of (cultured) bacteria samples was measured and evaluated by chemometric analysis of the THM-profiles. It was investigated whether the profiles obtained with two-dimensional GC resulted in more specific information for the identification of bacteria than one dimensional GC. Additionally, the presence of new, or recently proposed biomarkers was investigated. WGSM3: Dried Blood Spot sampling in early clinical development studies for pharmacokinetics, pharmacogenomics and safety assessments Jaap Wieling, Theo de Boer QPS Netherlands BV, Hanzeplein 1-53, 9713 GZ Groningen, The Netherlands For bioanalysis of drugs and also for (SNP)genotyping, dried blood spot (DBS) sampling has recently been reported several times as an alternative for venous sampling, mainly in preclinical research. Potential advantages include better estimation of pharmacological processes, stabilization of the analyte by the DBS matrix and storage without the need of freezers in less storage room. More over, the technique allows for less invasive sampling in non-clinical, i.e. ambulant, conditions. We investigated the usability of DBS for bioanalysis in a clinical phase I and bioanalytical environment using UHPLC-MS/MS, SNP genotyping and ICP-MS. WGSM4:


Adema, Leiden University

WGSM5: ZIC-HILIC as a 1st dimension in a two-dimensional liquid chromatography configuration provides high resolution separation and increased sensitivity in proteome analysis Serena di Palma In this presentation we introduce a 2D-LC strategy based on a combination of hydrophobic interaction (HILIC) and reversed phase (RP) chromatography. We compare two zwitterionic chromatographic materials (ZIC-HILIC and ZIC-cHILIC) as first dimension with respect to peptide separation efficiency and peptide/protein identification. Both approaches allowed the identification of thousands of proteins from a remarkably low amount of starting material (cells or tissue). The resulting data allows a comprehensive picture to be built regarding ZIC(c) peptide separation characteristics. We demonstrate such a method is both sensitive and possesses a high resolving power. WGSM6: Optimization of protein digestion for the characterization of drug-protein adducts Switzar L., Lingeman H., Irth H., Giera M. and Niessen W.M.A. VU University Amsterdam, Faculty of Sciences, BioMolecular Analysis Group, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands Drug-protein adducts are suggested to play a role as mediators of Adverse Drug Reactions (ADR's). Their early detection and identification is crucial for a successful drug


development. Several Liquid Chromatography-Mass Spectrometry (LC-MS) based strategies have been developed for the analysis of drug-protein adducts, most of which depend on enzymatic digestion of the modified protein followed by LC-MS(MS) analysis of the resulting peptides. In this respect, it is crucial to choose the most effective protease and apply the optimal digestion conditions, such as buffer pH, temperature and digestion time. In this presentation, we discuss an experimental design approach for the optimization of the digestion conditions for the enzymes trypsin, chymotrypsin and thermolysin. Two read out parameters are of importance: the yield of the peptide fragment containing the modification and the overall protein coverage. The found optima showed huge discrepancies to numerous published digestion conditions. Moreover, interactions between all studied variables could be revealed. WGSM7: Characterization of biopharmaceuticals by capillary electrophoresis ­ mass spectrometry Rob Haselberg*, Gerhardus J. de Jong and Govert W. Somsen Department of Biomedical Analysis, Utrecht University, P.O. Box 80082, 3508 TB Utrecht, the Netherlands. *E-mail: [email protected] With efficient methodologies available in biotechnology today, increasing numbers of recombinantly manufactured pharmaceutical peptides and proteins are being commercialized. The assessment of product quality is an essential issue during manufacturing. Furthermore, with the appearance of biosimilars the demand for characterization of biopharmaceuticals has even further increased. There is a growing need for suitable methods that not only allow identification, but also separation and quantification of impurities and possible degradation products. Capillary electrophoresis-mass spectrometry (CE-MS) provides the high separation efficiency and mass-selective detection that is required for biopharmaceutical product characterization. As CE has the intrinsic capacity to produce narrow peaks, it shows good potential for the separation of a variety of protein modifications in a single run. MS detection of high mass accuracy and resolution, such as provided by time-of-flight (TOF) instruments, yields highly useful information on the molecular weight of analyzed proteins species. In this presentation, the applicability of CE-TOF-MS for biopharmaceutical quality issues such as purity, stability, heterogeneity, and product composition will be outlined. To prevent protein adsorption, charged coatings were applied to the inner capillary wall, yielding efficient and reproducible protein separations. It will be shown that CE-TOF-MS allows separation and identification of degradation products resulting from prolonged storage and heat stress of the drugs recombinant growth hormone and oxytocin. The capacity of CE-MS to provide highly specific glycoform profiles of pharmaceuticals such as interferon--1a and erythropoietin will also be demonstrated. Compositional characterization of synthetic drug-protein products by CE-TOF-MS will be treated as well, showing that drug loading values can be obtained. DSP1: Characterization of maltodextrin ­ acrylic hybrid copolymers J. Purmová, W. Rutterkamp, L. Vertommen Measurement & Analytical Science (MA-CWA) Akzo Nobel Chemicals bv Research, Development & Innovation Zutphenseweg 10, P.O. Box 10 7400 AA Deventer, The Netherlands Many aqueous industrial systems require various materials to remain in a soluble, suspended or dispersed state. Often the water in those systems contains ingredients such as inorganic salts. These salts can cause accumulation, deposition, and fouling problems. Synthetic polymers have been used to minimize scale formation in aqueous treatment systems for a number of years. However, there has been a shortage of monomers to produce these synthetic polymers lately due to rising demand and tight crude oil supplies. Hence, there is a need to replace these synthetic polymers with hybrid polymers that are at least partially derived from renewable natural sources. Also, polymers from renewable natural sources should have a better biodegradable profile than synthetic polymers.


Hybrid copolymers consisting of (mostly) acrylic based copolymers attached onto the glucose units of maltodextrin (abbreviation MD; polymer of glucose having usually 2 ­ 20 glucose units) are produced at AkzoNobel Surface Chemistry in Chattanooga. The attachment occurs during the radical polymerization of acrylic monomers and the hybrids consist of maltodextrin backbone and acrylic side chains (Figure 1).


Unsubstituted Glucose units

Initiator, acrylic monomers Non-grafted acrylics

Hybrid copolymer Unsubstituted Maltodextrin chains Glucose units grafted with acrylic copolymer

Figure 1. Schematic representation of the synthesis and composition of hybrid copolymers To further development and tailoring of the properties for different applications good characterization of the hybrid co-polymers is necessary. Analytical methods for the characterization of hybrid maltodextrin-synthetics copolymers including different sample pre-treatment and chromatographic techniques were developed at ECG Measurement & Analytical Sciences in Deventer. With the aid of these methods the following properties could be determined: Amount of free maltodextrin Amount of unsubstituted glucose units Molecular weight distribution of hybrid copolymer and synthetics side chains Attempt to separate the mixture with the aid of GPEC (Gradient Elution Polymer Chromatography) and to quantify non-grafted acrylic polymers will be discussed as well DSP2: Automated (multivariate) data analysis for SEC Jos de Heer, SABIC Innovative Plastics Determination of the molecular weight distribution (MWD) of individual polymers in a polymer blend yields important information for product developers e.g. on stability during processing. The procedures of sample preparation and data analysis for SEC can be complex and time consuming. This presentation will discuss our automated workflow, which covers sample preparation, sample analysis, data treatment, MWD calculation and results upload into the laboratory database. These steps have been seamlessly integrated using MatlabTM algorithms developed in house. The data treatment is illustrated using real life examples of MWD analysis of polycarbonate/polyester and polyphenyleneoxide/polystyrene blends by using SEC/UV-DAD or SEC/RI in combination with data subtraction or multivariate data analysis through principal component regression. The automated analysis results in a higher productivity, lower operator workload, reduced turn-around-time and improved data consistency. DSP3: Protein Analysis by Size Exclusion Chromatography Greg Saunders, Agilent Technologies


The analysis of proteins by size exclusion chromatography (SEC) is an area of growing interest. SEC is a well-known technique for the analysis of macromolecules and involves the separation of molecules on the basis of size in solution, a property that can be related to molecular weight. The analysis of proteins by SEC is advantageous as it allows the purity of proteins to be assessed and the presence of dimmers, trimers and higher polymers to be confirmed, as these species can inhibit therapeutic activity. However, proteins are considerably more complex than most synthetic macromolecules, and protein SEC is more difficult to perform routinely. This presentation will discuss the pitfalls of protein analysis by SEC, and highlight some of the advances that have been made in this challenging field. DSP4: Using an Evaporative Light scattering Detector (ELSD) for quantification with LC, a blessing or a curse? Bastiaan Staals BASF 2-Dimensional chromatography (HPLC-SEC) requires that the eluent from the 1st dimension is compatible with the SEC column. This is not always the case. A way to circumvent this is off line fractionation. Fractions are taken from the 1st dimension and dried. Afterwards, dissolved in a SEC compatible solvent and measured with SEC. Despite the fact that this is somewhat more laborious, this method has some major draws backs. A few here are mentioned: In order to reconstruct the original chromatogram in SEC, the fractions should be re-dissolved with an amount of solvent that is directly related to the volume in which these fractions were collected. (If one takes constant time = volume intervals from the first dimension this problem circumvented).High boiling solvents like DMAc are hard to remove, even under vacuum. Using temperature and high vacuum might also modify the fractions (chemically instable). Often a pre-concentration is needed. This makes the handling with low volumes complicated. The new generation of ESLD detectors has increased dramatically in terms of sensitivity. In this way there is no need for pre-concentrating the samples for SEC. However, solving one problem often creates another one. If one would like to quantify with ELSD detectors like Corona, not all of it is straight forward. In this lecture I'll hope to shine some light on the pro's and contras of quantifying polymer mixtures with these detectors. DSP5: Industrial applications of SEC hyphenated with ESI-TOF-MS Paul Cools, Ton Brooijmans DSM NeoResins+, Waalwijk, The Netherlands With the growing safety awareness in the industry in general and the introduction of R.E.A.C.H. the need to analyse low molar mass species in polymers has become more evident. With size exclusion chromatography (SEC) low molar mass species can be separated and with the use of mass spectrometry, specifically electrospray time of flight mass spectrometry (ESI-TOF-MS), the separated species can be identified based on accurate mass. Some application of SEC-MS will be presented in comparison with gradient LC-MS (applied on the same hardware). Furthermore, practical aspects of the SEC-MS system, and especially interpretation issues will be discussed. DSP6: Challenges in molecular weight determination of PEG-pNIPAm block copolymers Albert J. de Graaf, Kristel W.M. Boere, Mies J. van Steenbergen, Wim E. Hennink Department of Pharmaceutices, Utrecht Institute of Pharmaceutical Sciences, Universiteit Utrecht, Utrecht, the Netherlands In our department we are studying polymeric micelles as delivery vehicles for hydrophobic, low molecular weight drugs. Micelles with a hydrophobic core and a


hydrophilic shell are obtained by dispersing amphiphilic AB blockcopolymers in water, whereas BAB triblock copolymers with a hydrophilic midblock flanked by two hydrophobic blocks have been hypothesized to self-assemble into so-called `flower-like micelles'. In order to compare the behavior of such `flower-like micelles' and their `conventional' counterparts, we prepared AB and BAB block copolymers of poly(ethylene glycol) (PEG), block A, and the thermosensitive poly(N-isopropylacrylamide) (pNIPAm), block B. A series of diblock and triblock copolymers with varying pNIPAm length was prepared by Atom Transfer Radical Polymerization (ATRP). When trying to exclude the presence of diblock copolymers in our triblock copolymer preparations, we encountered several difficulties in Gel Permeation Chromatography (GPC). First of all there are the difficulties associated with GPC of any block copolymer, namely the dependence of hydrodynamic volume and refractive index increment (dn/dc) on the ratio of block lengths. Secondly, formation of branched or cyclic chains in ATRP cannot be a priori excluded. Main problem however is the capability of poly(acrylamides) such as pNIPAm to form persistent inter- and intra-chain hydrogen bonds and metal complexes, which consistently lead to an overestimation of their molecular weight in GPC. In this contribution the effectiveness of several approaches to minimize these difficulties will be presented, such as sample preparation, concentration, GPC eluent and temperature. Furthermore, it will be shown that even `absolute' molecular weight determination with viscometry, Static Light Scattering (SLS) and Mass Spectrometry (MALDI-TOF MS) are not trouble-free. To end with a positive note, however, one of our recent experiments will be described in which we actually took advantage of the aforementioned overestimation of pNIPAm molecular weight in GPC. DSP7: Novel pyrolysis GC-MS methods for studying the dissolution behaviour of polymeric materials in aqueous media Aleksandra Chojnacka, Hans-Gerd Janssen and Peter Schoenmakers Analytical-Chemistry Group, Faculty of Science, University of Amsterdam, Nieuwe Science Park 904, 1098 XH Amsterdam, The Netherlands ([email protected]) Knowledge on the solubility and dissolution behaviour of polymeric materials has become more important during the last decade, especially for polymer science and engineering. This is due to the increasing number of applications of polymers in industry and society. In the medical field typical examples are polymeric implants and controlled-release media for drug delivery, be it for implantation under the skin, injection into the body, or oral intake. For these uses knowledge of the rates of dissolution and/or extraction is vital. This is also the case in other applications of specialty or commodity polymers, for example as food-packaging materials. Because polymers can be extremely complex in terms of molecular weight (distribution), chemical composition, end groups, etc., and because dissolution and extraction are often meant to be (very) slow, accurate rate measurements are highly challenging. In this project we are developing methods for the qualitative (identification and determination of composition) and quantitative analysis of dissolved copolymers in water. The methods are based on pyrolysis coupled to capillary GC-MS. Adequate sensitivity of the methods is ensured by liquid injection of large volumes, typically 10 to 20 L. The conditions for thermal conversion with or without in-situ methylation are optimized. Cryofocussing on top of the column is employed to enhance sensitivity. Copolymers of vinylpyrrolidone and vinylacetate are of interest because of their good binding and film-forming properties, affinity to hydrophilic and hydrophobic surfaces, and relatively low hygroscopicity. Py-GC-MS was used to study commercially available products of this nature. The pyrolysis conditions were optimized and the quantitative aspects of the method were studied.


Put up your poster at the indicated place


First author





Gel-free proteomics of covalently attached Bacillus subtilis spore coat proteins

Abhyankar, W.R.

W. R. Abhyankar, A. Ter Beek, H. L. Dekker, S. Brul and C. G. de Koster

University of Amsterdam


An Analysis Framework for Complex Data Sets from Various Experimental Sources

Ahmad, I.

I. Ahmad, George Byelas, M. Dijkstra, M. Swertz, F. Suits, B. van Breukelen, R. Bischoff, P. Horvatovich

Wageningen University and Research Centre Radboud University Nijmegen Wageningen University and Research Centre



Using 19F NMR for structure elucidation

Ampt, K.A.M.

K.A.M. Ampt, M. Jeager, P.E.T.J. Geutjes, S.S. Wijmenga and M. Honing

Institute of Molecules and Materials


Micro-bioaffinity mass spectrometry of mycotoxins in wheat

Aqai, Payam

Payam Aqai, Arjen Gerssen, Willem Haasnoot, Linda Stolker, Hans Mol, Michel Nielen



Microbial peptidomics

Bener-Aksam, E.

E. Bener-Aksam, M.W.H. Pinkse and P.D.E.M. Verhaert.

Delft University of Technology

Analytical Biotechnology


The discrimination of eight chloramphenicol isomers by LC-MS/MS

Berendsen, Bjorn

Bjorn Berendsen, Linda Stolker, Michel Nielen

Wageningen University and Research Centre



Metabolic profiling of processed fruits and vegetables

Bialek, Lucy

Patricia Lopez-Sanchez(1), John van Unilever Discover Duynhoven (1), Lucy Bialek(1), Robert Vlaardingen Hall (2), Ric de Vos(2)



Characterization and inhibition of Hsp90 by Circular Dichroism and Surface Plasmon Resonance.

Binchi, Laura

Laura Binchi,b, N.J. de Molb, M.J.E. Fischerb, Jongb, C.Bertuccia Lionel Blanchet1, Agnieszka Smolinska1, Amos Attali2, Therese Rosenling3, Marcel Stoop4, Leon Coulier5, Marek Noga6, Shanna Shi6, Adrie Dane6, Kirsten Ampt1, Tinka T. G. Bloemberg1, G. F. Giskedegrd2, L. M. C. Buydens1, G. Postma1, B. Sitter2, I. S. Gribbestad2, and T. F. Bathen2 W.P.H. de Boer, J.Lankelma, R. Bischoff, P. Horvatovich

Utrecht University

bDepartment of Pharmaceutical Sciences


Mid level fusion in -omics: application in biomarkers discovery of multiple sclerosis

Blanchet, Lionel

Radboud University Nijmegen Radboud University Nijmegen

IMM/Analytical Chemistry


Peak Alignment of HR-MAS NMR spectra

Bloemberg, T.G.

IMM / analytical chemistry


Program Curfit: two-dimensional alignment of LCMS chromatograms

Boer, W.P.H. de

VU University Medical Center

Medical Oncology

UPLC-tandem MS analysis of amino acids in 12 lysozyme and lysozyme hydrolysate formulations containing carbohydrates

Boogers, Ilco

Ilco Boogers, Pieter Stam, Alexander L.L. Duchateau,




Profiling HMTD using Isotope-Ratio Mass Spectrometry

Brust, Hanneke

Hanneke Brust, Peter Schoenmakers, Mattijs Koeberg, Martin van Breukelen, Arian van Asten Salvatore Cappadona, Linda Pattini, Fredrik Levander, Reinout Raijmakers, Shabaz Mohammed, Albert Heck and Bas van Breukelen A.Chojnacka, A Ghaffar, P.Schoenmakers

University of Amsterdam

zowel via prof dr de koster als via abhyankar Biomolecular MS and Proteomics


Wavelet-based approach for optimal denoising of HPLC-MS data

Cappadona, Salvatore

University of Utrecht


Pyrolysis GC-MS methods for studying the composition and dissolution behaviour of copolymers Development of a Glass Microfluidic Device and Incubation System for Human Umbilical Vein Endothelial Cell (HUVEC) Culture Under Flow Conditions

Chojnacka, A.

University of Amsterdam



Dijk, Maurits van

Maurits van Dijk, Patty P.M.F.A. Mulder, Elisabeth Verpoorte

University of Groningen

Pharmaceutical Analysis



Analysis of the ATPase cycle of full length Hsp90 by NMR

Duarte, Afonso

Afonso Duartea, Elif Karagöza, Hans Ippelb, Martijn van Rosmalena, Rolf Boelensb and Stefan Rüdigera Esther van Duijn1, Arjan Barendregt1, Jelle B. Bultema2, Matthijs M. Jore3, Magnus Lundgren3, Stan J. Brouns3, Blake Wiedenheft4, Jennifer A. Doudna4, Egbert J. Boekema2, John Voda1, A., Homan2, N., van Dalen1, G. van, Nijsse1, J., van As2, H., van Duynhoven1, J.

Utrecht University

Bijvoet Center for Biomolecular Research, Biomolecular mass spectrometry and proteomics group


Structural characterization of CRISPR-RNA antiviral defense systems by tandem- and ion mobility mass spectrometry

Duijn, Esther van

Utrecht University


Multiscale characterization of microstructure and rehydration behavior of dried carrots

Duynhoven, J. van

1Unilever Discover Vlaardingen Analytical Biotechnology Group & Netherlands Proteomic BioMolecular Analysis Group

A novel strategy to rapidly discover cysteine rich 20 peptides from crude frog secretions, their primary structure and biological activity On-line modification, identification and biological characterization of p38 kinase inhibitors by electrochemical oxidation coupled to a hyphenated screening assay

Evaristo, Geisa

Geisa P.C. Evaristo1*, Martijn Pinkse1, Tianbao Chen2, Chris Shaw2, Peter Verhaert1 Falck D.1, de Vlieger J.S.B.1, Giera M.1, Kool J.1, Honing M.2, Niessen W.M.A.1 and Irth H.1 Lorenza Franciosi, Natalia Govorukhina, Nick ten Hacken*, Dirkje Postma*, Fabrizia Fusetti°, Bert Poolman°, Rainer Bischoff



Falck, D.

VU University Amsterdam


Biomarker discovery in COPD using Epithelial Lining Fluid: a proteomic approach

Franciosi, Lorenza

University of Groningen

Department of Pharmacy Biomolecular Mass Spectrometry and Proteomics Groups Analytical Chemistry Group, HIMS, FNWI


Peptide sequencing by electron transfer dissociation and beam type collision induced dissociation for improved peptide and protein identification

Frese, C.

C. Frese, A.F.N. Altelaar, S. Mohammed, A.J.R. Heck

Utrecht University

Monitoring the in vitro degradation of degradable 24 poly(ester amide)s for controlled drug delivery by HPLC-TOF-MS

Ghaffar, A.

A. Ghaffar, G. Draaisma, G. Mihov, A. A. Dias, P.J. Schoenmakers, Sj. van der Wal

University of Amsterdam


Assessment of the Specific Surface Area of fat crystal networks by Diffusion NMR

Goudappel, Gert-Jan

Gert-Jan W. Goudappel, Jaap Nijsse

Unilever R&D Vlaardingen

Laser microdissection and cervix carcinoma Govorukhina, 26 proteomics : spatial cellular resolution of four heat Natalia shock biomarker candidates



Natalia I. Govorukhina, Coskun Guzel, Klaske. A. ten Hoor, Harry G. Klip, Ate G.J. van der Zee, Theo M. Luider and Rainer Bischoff Chandrakala Gowda, Erik Schwartz, Filipe Vasconcelos, Ernst RH Van Eck, Gowda, Sander Weezenberg, Gilles de wijs, Chandrakala Georg Kresse, Martijn Marsman, Jeroen JLM Cornelissen, Roeland JM Nolte, Greiderer, Andreas Andreas Greiderer, Gabriel Viv-Truyols, Linda Steeneken, Tom Aalbers, Peter Schoenmakers

University of Groningen

Analitical Biochemistry

Radboud University Nijmegen

Institute for Molecules and Materials Vant Hoff Institute for Molecular Sciences Unit Research and Development


Characterization of Hydroxypropylmethylcelllose using Comprehensive Two-Dimensional Liquid Chromatography

University of Amsterdam


NMR artifacts during quantitative analysis

Hamzink, Martin

Martin Hamzink and Bert Zomer

The Netherlands Vaccine Institute


Evaluation of Performance and Benefit of UltrahighHeijden, Rob Resolution ESI-TOFMS Coupled to Fast van der Chromatography in the Application of Forensic Screening Hennrich, Marco L.

Rob van der Heijden1, Patrick van Bruker Nederland Houts1, Anna Pelander2, James Hillis3 BV, and Petra Decker4 Biomolecular Mass Spectrometry and Proteomics Group Analytical Biochemistry

Dimethyl isotope labeling assisted de novo peptide 31 sequencing

Marco L. Hennrich, Shabaz Mohammed, A.F. Maarten Altelaar, Albert J.R. Heck

University of Utrecht


The effect of surface tension on the electrospray response of N-acylated amino acids.

Hermans, Jos

Jos Hermansa, Sara Ongaya, Nicolas Abelloa, Paul Geurinkb, Herman Overkleeftb, Rainer Bischoffa

University of Groningen


Development of a miniaturized fluorescence detection system for the hyphenation of nano-LC to on-line biochemical assays

Heus. F.

Heus F., Giera M., Lingeman H., Kool J., Irth H. and Niessen W.M.A.

VU University Amsterdam

BioMolecular Analysis Group


msCompare ­ data processing framework for the analysis of label-free quantitative LC-MS data

Hoekman, B.

B.hoekman, P.horvatovich, R.bischoff

University of Groningen



Fluorescence study of the binding of antihistamine to human serum albumin

Hooijschuur, Jan-Hein

Jan-Hein Hooijschuur, Silvia Tardioli, Gert van der Zwan, Cees Gooijer Patrick van Houts1, Rob van der Heijden1, Peter Sander2, Ilmari Krebs2, Sebastian Goetz2, Birgit Schneider2 and Aiko Barsch2 Patrick van Houts1, Rob van der Heijden1, Andreas Brekenfeld2, Christoph Gebhardt2, Ralf Hartmer2, Thorsten Ledertheil2, Michael Schubert2 and Arnd Ingendoh2 Ingeborg E. Iping Petterson, Patrick Dvoák, Joost B. Buijs, Cees Gooijer and Freek Ariese

Vrije Universiteit Amsterdam

Biomolecular Analysis & Spectroscopy


Deconvolution and Automatic Formula Assignment Houts, Patrick of Alternating MS and Broad-Band CID Analyses van

Bruker Nederland BV


Pushing toward next generation Ion-Trap-MS: ion Houts, Patrick formation, transfer and trapping van

Bruker Nederland BV


Time-Resolved Spatially Offset Raman Spectroscopy for depth analysis of diffusely scattering layers

Iping Petterson, Ingeborg

Vrije Universiteit Amsterdam

Biomolecular Analysis & Spectroscopy


Artifacts in the analysis of sterol(ester)s in cholesterol lowering food products

Janssen, HansGerd

Hans-Gerd Janssen, Raymond Baris and Herrald Steenbergen M. Jupin, Nijmegen, The Netherlands, P. Michiels, F. Girard, Spinnovation, The Netherlands, M. Spraul, Bruker Biospin, Germany Kloos D.1, Giera M.1, Wijtmans M.2, Derks R.J.E.3, Mayboroda O.A.3, Deelder A.M.3, Irth H.1 and Niessen W.M.A.1 Dang Thi Ngoc A, Arend Kolk, Sjoukje Kuijper, Hans-Gerd Janssen

University of Amsterdam

AnalyticalChemistry Group


Identification and characterisation of metabolites binding to proteins in blood plasma Development of a pre-column derivatization strategy for the measurement of the tricarboxylic acid cycle intermediates by means of reversed phase LC-MS with ESI+ ionization Novel approaches in diagnosing tuberculosis in children

Jupin, M.

Radboud University


Kloos, D.

VU University Amsterdam

BioMolecular Analysis Group Analytical Chemistry & Forensic Science Institute of Molecules and Materials (IMM


Kolk, Arend

University of Amsterdam


Regioselective hydroxylation of Testosterone by drug-metabolising mutants of Cytochrome P450BM3 as studied by NMR relaxation

A.J. Kolkman1, V. Rea2, E.V. Vottero2, K.A.M. Ampt1, M. Tessari1 J.N.M. Kolkman, A.J. Commandeur2, H. Irth3, N.P.E. Vermeulen2, M. Honing4 and S.S. Wijmenga1 M.J. Lopez-Martinez, J.Vila-Planas, S. Demming, P.P.M.F.A. Mulder, S. Büttgenbach, A. Llobera, E. Verpoorte

Radboud University Nijmegen


Multiple Internal Reflection Poly(dimethylsiloxane) Lopezsystems for on-line pH monitoring in ultra-small Martinez, M.J. volumes

University of Groningen

Pharmaceutical Analysis group


Osteocalcin flow cytometric immunoassay as a tool to detect rbST abuse in cattle

Ludwig, Susann

Susann Ludwig, Maria G.E.G. Bremer, Nathalie G.E. Smits, Michel W.F. Nielen

Wageningen University and Research Centre



Spatially-offset Raman spectrometry

Mank, A.J.G.

A.J.G. Mank, E.J.K. Verstegen

Philips Research

Materials Analysis


Stability-Indicating Analysis of the Acetylcholinesterase Inhibitors (-) Huperzine A, Tacrine and Galantamine

Marques, L.A.

Marques L.A.1, Maada I.1, Giera M.1, Kool J.1, de Kanter F.J.J.2, Lingeman H.1, Niessen W.M.A.1 and Irth H.1 Heleen Meuzelaar1, Michal Heger2,3, Pilar de la Torre1, Marleen M. Kerssens1, Gert van der Zwan1 Paul M. van Midwoud, Marjolijn T. Merema, Elisabeth Verpoorte, Geny M.M. Groothuis

VU University Amsterdam

BioMolecular Analysis Group Department of Biomolecular Analysis & Spectroscopy Pharmaceutical Analysis


Elucidation of the spectroscopic behavior of Adenosine-5'-triphosphate in aqueous solution

Meuzelaar, Heleen

VU University Amsterdam


University of Groningen


Time alignment strategy for LC-MS data obtained in an inter-laboratory study

Mitra, V.

Mitra V. 1, Bischoff R. 1, Smilde A. 2, Hoefsloot H. 2, Horvatovich P. 1

University of Groningen

Department of Analytical Biochemistry


On-line, salt-free, two-dimensional nanoflow-LCMS for comprehensive epitope-mining

Mommen, Geert

Ad de Jong; Geert Mommen


Analytical research



Mulder, Patty

Patty P.M.F.A. Mulder and Elisabeth Verpoorte

University of Groningen




Development of CSF metabolomics for biomarker discovery in neurological research

Noga, M.J.

Noga M.J., Shi S., Guled F., Dane A., Attali A., Tuinstra T., Coulier L., Leiden University Reijmers T.H., Vreeken R.J., Luider T., Hankemeier T. Jos Hermans, Sara Ongay, Nicolas Abello, Paul Geurink, Herman Overkleeft, Rainer Bischoff University Medical Center Groningen Analytical Biochemistry


The effect of surface tension on the electrospray response of N-acylated amino acids.

Ongay, Sara


Miniaturised chip-based devices for liquid/liquid extraction and sample injection in capillary gas chromatography: theory and applications.

Peroni, Daniela

Daniela Peroni, Wil van Egmond, HansGerd Janssen

University of Amsterdam


Metabolite discovery through trend detection in multi-dose kinetic studies

Peters, Sonia

Sonja Peters, Hans-Gerd Janssen, Gabriel Vivo-Truyols

University of Amsterdam


Visualization and Recovery of the (Bio)chemical 57 Interesting Variables in Data Analysis with Support Postma, G.J. Vector Machine Classification

L.M.C. Buydens1, P.W.T. Krooshof1, B.stn2, and G.J. Postma1

Radboud University Nijmegen

IMM/Analytical Chemistry


Activity-based profiling and quantification methods for MMP9 and MMP12

Prelya, Lauretta M.

U, Theo Kleina, Krisztina Paala, Erwin Tuinb, Herman S. Overkleeftb, Antoon J.M. van Oosterhoutc, Rainer Bischoffa

University of Groningen

Analytical Biochemistry


Combined determination of cholesterol and triglycerides in serum lipoproteins with AsFlFFF coupled to THM-GC-MS

Qureshi, Rashid Nazir

Rashid Nazir Qureshi, Erwin Kaal, Wim Th. Kok, Peter J. Schoenmakers

University of Amsterdam



Polar metabolite quantification in complex matrixes using GC-MS

Ries, Marco

Marco Ries, Rob Vreeken, Thomas Hankemeier

Leiden University

LACDR/Analytic al Biosciences


Affinity Enrichment of Glycoproteins from serum of Cervical Cancer patients

Robin, I.J.

I.J. Robin, G. Carlucci, N. Govorukhina, P. Horvatovich, V. Mitra, A. van der Zee and R. Bischoff Niels de Roo, Imane Mahlous, Monique Klinkenberg, Pauline Alexandre, Doris Jacobs, Hans-Gerd Janssen, John van Duynhoven Therese Rosenling, Marcel Stoop, Amos Attali, Christin Christin, Frank Suits, Peter Horvatovich, Tinka Tuinstra, Theo Luider, Rainer Bischoff Saccenti, E., van Eeuwijk F. A. , Szymaska, E. , Strassburg, K. , Dane, A. Saris, W. H. M. , van Duynhoven, J. P. , Smilde, A. K.

University of Groningen

Analytical Biochemistry group


Towards identification of polyphenol metabolites in Roo, Niels de bodyfluids by SPE1-LC-SPE2-cryoNMR-tofMS

Unilever R&D Vlaardingen



Proteins in cerebrospinal fluid (CSF) as biomarker candidates for experimental autoimmune encephalomyelitis (EAE) in rats

Rosenling, Therese

University of Groningen

Analytical Chemistry


Optimal measurement designs: a forgotten topic in metabolomics studies

Saccenti, E.

University of Amsterdam



Using TERS to monitor a heterogeneous catalytic reaction in-situ

Schrojenstein Evelien M. van Schrojenstein Lantman, Lantman, Tania Deckert-Gaudig, Arjan J.G. Mank, Evelien van Volker Deckert, Bert M. Weckhuysen J.M. Posma1, A. Smolinska1, A. Attali2, A. van Gool3, T. Luider4, S.S. Wijmenga1 and L.M.C. Buydens1

Utrecht University

Debye Institute for Nanomaterials Science Institute for Molecules and Materials (IMM) Institute for Molecules and Materials (IMM)


Data fusion of 1H-NMR metabolic spectra

Smolinska, A.

Radboud University Nijmegen Radboud University Nijmegen Radboud University Nijmegen Eindhoven University of Technology

R. ter Horst1, A. Smolinska1, A. Atali2, Multivariate analysis of 2D 1H NMR spectra of preP. Michiels3, F. Girard3, T. Luider4, A. Smolinska, A. 67 clinical model of Multiple Sclerosis van Gool5, S.S. Wijmenga1 and L.M.C. Buydens1 Multivariate analysis for biomarker selection in Multiple Sclerosis A. Smolinska, K. Ampt. T. Luider, A. van Gool, S. Wijmenga, L. Buydens


Smolinska, A.

IMM / analytical chemistry


Mg based metal hydrides for hydrogen storage. A NMR perspective

Srinivasan, Subramanian

Subramanian Srinivasan, Pieter C.C.M.Magusin, Rutger A. van Santen, Peter H.L. Notten.

Department of Chemical Engineering



Morphological analysis of MRI for the discrimination of brain tumors.

Stens, Erik

Lionel Blanchet1, Erik Stens1, Geert Postma1, Albert Idema2, Arend Heerschap3 and Lutgarde Buydens1

Radboud University Nijmegen

Institute for Molecules and Materials (IMM)


Optimization of protein digestion for the characterization of drug-protein adducts

Switzar, L.

Switzar L., Lingeman H., Irth H., Giera M. and Niessen W.M.A.

VU University Amsterdam

BioMolecular Analysis Group


Very fast and efficient size-based separations of polymers at ultra-high pressures

Uliyanchenko, E.

E. Uliyanchenko, P.J. Schoenmakers, Sj. van der Wal

University of Amsterdam



Construction of a spatial-chromatographic device containing a flat and wide monolithic stationary phase: flow-profile investigation Resonance Raman spectroscopic characterization of Indoleamine 2,3 dioxygenase complexed with menadione Molecular conformation and ordering of amphiphilic peptides by rotational-echo doubleresonance NMR

Vanhoutte, Dominique

Dominique Vanhoutte; Johan E.M. Soede; Peter J. Schoenmakers; Wim Th. Kok Cecilia Vidami Negoescu, Eduardo Vottero§, Cees Gooijer, Gert van der Zwan Villanueva-Garibay, J. A., van den Heuvel, M., Löwik, D. W. P. M., Kentgens, A. P. M.

University of Amsterdam



Vidami Negoescu, Cecilia

Vrije Universiteit

Biomolecular Analysis and Spectroscopy


VillanuevaGaribay, J.A.

Radboud University Nijmegen

Department of Solid-State NMR

Towards sub-typing of rheumatoid arthritis H.A. Van Wietmarschen, T.H. Reijmers, Wietmarschen 76 patients using a questionnaire based on a fusion of K. Wei, J. Schroën, J.J. Meulman, J. Leiden University , H.A. van van der Greef Chinese and Western diagnosis A new approach for generic screening and A.M. van Wijk1, B. Beerman1, H.A.G. quantitation of potential genotoxic alkylation Wijk, A.M. van Niederländer 1, A.H.G. Siebum1, G.J. agents by pre-column derivatization and LCMS/MS de Jong 2 analysis Phospholipidomic identification of potential biomarkers of Diabetic nephropathy and Diabetic mellitus Chao Zhu, Qionglin Liang, Heng Wei, Mei Wang, Jan van der Greef Abbott Healthcare Products BV

Analytical Biosciences


Analytical Development


Zhu, Chao

Leiden University

Division of Analytical Bioscience




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