of The Association for Mass Spectrometry · The Association for Mass Spectrometry: Applications to the Clinical Lab Salzburg, AUSTRIA September 9 - 13, 2018 Salzburg Congress Center

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MSACL 2018 EU

The 5th Annual European Congress of

The Association for

Mass Spectrometry: Applications to the Clinical Lab

Salzburg, AUSTRIA

September 9 - 13, 2018 Salzburg Congress Center

The Association is a non-membership, non-profit 501(c)(3) tax-exempt California Corporation

with the mission of furthering education in the field of mass spectrometry.

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Table of Contents

SPONSORSHIP & TRAVEL GRANT SUPPORT ............................................................................................................5 SCIENTIFIC COMMITTEE ..........................................................................................................................................6 GENERAL INFORMATION ...................................................................................................................................... 16 PRESENTER INFO AND GUIDELINES ...................................................................................................................... 17 SCHEDULE OVERVIEW .......................................................................................................................................... 18 PLENARY & KEYNOTE SPEAKER SERIES ................................................................................................................. 21 YOUNG INVESTIGATOR GRANTS ........................................................................................................................... 26 LAB DIRECTOR GRANTS ........................................................................................................................................ 28 TRAINEE GRANTS.................................................................................................................................................. 28 SHORT COURSE OVERVIEW .................................................................................................................................. 29 EXHIBITS SUMMARY ............................................................................................................................................. 31 EXHIBITORS .......................................................................................................................................................... 32 CORPORATE WORKSHOPS ..................................................................................................................................... 35 PODIUM PRESENTATIONS .................................................................................................................................... 39

• SESSION 1 • TRACK 1 • LIPIDOMICS WEDNESDAY @ 9:00 IN MOZART 1-3 SESSION CHAIR: JULIJANA

IVANISEVIC - UNIVERSITY OF LAUSANNE .............................................................................................................. 40 • SESSION 1 • TRACK 2 • DISEASE IMPLICATIONS AND DIAGNOSIS WEDNESDAY @ 9:00 IN MOZART 4-5

SESSION CHAIR: GRACE VAN DER GUGTEN - ST PAUL’S HOSPITAL ............................................................................. 41 • SESSION 1 • TRACK 3 • PROTEOMICS KEYNOTE WEDNESDAY @ 9:00 IN PAPAGENO HALL SESSION CHAIR: CHRISTA COBBAERT - LEIDEN UNIVERSITY MEDICAL CENTER ................................................................................... 42 • SESSION 1 • TRACK 4 • NEW TECHNOLOGIES WEDNESDAY @ 9:00 IN PARACELSUS HALL SESSION CHAIR: VLADIMIR FRANKEVICH - NATIONAL RESEARCH CENTER FOR OBSTETRICS AND GYNECOLOGY ......................................... 43 • SESSION 1 • TRACK 5 • ENDOCRINOLOGY WEDNESDAY @ 9:00 IN TRAKL HALL SESSION CHAIR: FLAMINIA

FANELLI - UNIVERSITY OF BOLOGNA ................................................................................................................... 44 • SESSION 1 • TRACK 6 • PRACTICAL TRAINING : PROTEOMIC ASSAY DEVELOPMENT WEDNESDAY @ 9:00 IN

DOPPLER HALL SESSION CHAIR: TIM COLLIER - QUEST DIAGNOSTICS - CLEVELAND HEART LAB ...................................... 45 • SESSION 2 • TRACK 1 • METABOLOMICS METHODOLOGY WEDNESDAY @ 11:00 IN MOZART 1-3 SESSION

CHAIR: ELIZABETH WANT - IMPERIAL COLLEGE LONDON ......................................................................................... 46 • SESSION 2 • TRACK 2 • ALL ABOUT BREATH ANALYSIS WEDNESDAY @ 11:00 IN MOZART 4-5 SESSION

CHAIR: NEUS FABREGAT-CABELLO - UNIVERSITY OF LIEGE ...................................................................................... 47 • SESSION 2 • TRACK 3 • ENDOCRINE KEYNOTE WEDNESDAY @ 11:00 IN PAPAGENO HALL SESSION CHAIR: BRIAN KEEVIL - MANCHESTER UNIVERSITY ........................................................................................................... 48 • SESSION 2 • TRACK 4 • GLYCOMICS 1 WEDNESDAY @ 11:00 IN PARACELSUS HALL SESSION CHAIR: GUINEVERE LAGEVEEN-KAMMEIJER - LEIDEN UNIVERSITY MEDICAL CENTER ............................................................... 49 • SESSION 2 • TRACK 5 • MS/MS FOR PROTEOTYPING PATHOGENS WEDNESDAY @ 11:00 IN TRAKL HALL

SESSION CHAIR: SIMON CAMERON - IMPERIAL COLLEGE LONDON ............................................................................ 50 • SESSION 2 • TRACK 6 • PRACTICAL TRAINING : CALIBRATORS AND INTERNAL STANDARDS FOR PROTEIN MS

WEDNESDAY @ 11:00 IN DOPPLER HALL SESSION CHAIR: CHRIS SHUFORD - LAB CORP .............................................. 51 • SESSION 3 • TRACK 1 • METABOLIC MARKERS WEDNESDAY @ 14:30 IN MOZART 1-3 SESSION CHAIR: LUKE

WHILEY - IMPERIAL COLLEGE LONDON ................................................................................................................ 52 • SESSION 3 • TRACK 2 • KEYNOTE SMALL MOLECULES WEDNESDAY @ 14:30 IN MOZART 4-5 SESSION CHAIR: MICHAEL VOGESER - MEDICAL UNIVERSITY OF MUENCHEN .................................................................................... 53 • SESSION 3 • TRACK 3 • CARDIOVASCULAR AND THROMBOTIC DISEASE WEDNESDAY @ 14:30 IN PAPAGENO

HALL SESSION CHAIR: TIM COLLIER - QUEST DIAGNOSTICS - CLEVELAND HEART LAB .................................................... 54 • SESSION 3 • TRACK 4 • TISSUE IMAGING KEYNOTE WEDNESDAY @ 14:30 IN PARACELSUS HALL SESSION

CHAIR: TIFFANY PORTA - MAASTRICHT UNIVERSITY ............................................................................................... 55 • SESSION 3 • TRACK 5 • GLYCOMICS 2 WEDNESDAY @ 14:30 IN TRAKL HALL SESSION CHAIR: TBD ........................... 56 • SESSION 3 • TRACK 6 • PRACTICAL TRAINING : EU REGULATIONS WEDNESDAY @ 14:30 IN DOPPLER HALL

SESSION CHAIR: BRIAN KEEVIL - UNIVERSITY OF MANCHESTER ................................................................................ 57

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• SESSION 4 • TRACK 1 • METABOLOMICS – CLINICAL STUDIES I THURSDAY @ 9:00 IN MOZART 1-3 SESSION

CHAIR: OLEG MAYBORODA - LEIDEN UNIVERSITY MEDICAL CENTER ......................................................................... 58 • SESSION 4 • TRACK 2 • QUALITY ISSUES THURSDAY @ 9:00 IN MOZART 4-5 SESSION CHAIR: ANNE BENDT -

NATIONAL UNIVERSITY OF SINGAPORE ................................................................................................................ 59 • SESSION 4 • TRACK 3 • CLINICAL CHEMISTRY I THURSDAY @ 9:00 IN PAPAGENO HALL SESSION CHAIR: ÉVA

HUNYADI-GULYÁS - BIOLOGICAL RESEARCH CENTER .............................................................................................. 60 • SESSION 4 • TRACK 4 • MSI FOR CLINICAL APPLICATIONS THURSDAY @ 9:00 IN PARACELSUS HALL SESSION

CHAIR: RAF VAN DE PLAS - DELFT UNIVERITY OF TECHNOLOGY ............................................................................... 61 • SESSION 4 • TRACK 5 • KEYNOTE FOR MICROBIOLOGY THURSDAY @ 9:00 IN TRAKL HALL SESSION CHAIR: JEAN ARMENGAUD - CEA-MARCOULE ............................................................................................................... 62 • SESSION 4 • TRACK 6 • PRACTICAL TRAINING : FINDING YOUR ANALYTE THURSDAY @ 9:00 IN DOPPLER HALL

SESSION CHAIR: CATARINA HORRO-PITA - LGC .................................................................................................... 63 • SESSION 5 • TRACK 1 • METABOLOMICS CLINICAL STUDIES II THURSDAY @ 11:00 IN MOZART 1-3 SESSION

CHAIR: NICOLA GRAY - UNIVERSITY OF READING .................................................................................................. 64 • SESSION 5 • TRACK 2 • QUANTITATION ISSUES THURSDAY @ 11:00 IN MOZART 4-5 SESSION CHAIR: CHRISTIANE AURAY-BLAIS - UNIVERSITE DE SHERBROOKE ....................................................................................... 65 • SESSION 5 • TRACK 3 • CLINICAL PROTEOMICS II THURSDAY @ 11:00 IN PAPAGENO HALL SESSION CHAIR: RENEE RUHAAK - LEIDEN UNIVERSITY MEDICAL CENTER......................................................................................... 66 • SESSION 5 • TRACK 4 • GLYCOMICS KEYNOTE THURSDAY @ 11:00 IN PARACELSUS HALL SESSION CHAIR: MANFRED WUHRER - LEIDEN UNIVERSITY MEDICAL CENTER .................................................................................. 67 • SESSION 5 • TRACK 5 • TECHNOLOGICAL INNOVATIONS FOR THE MICROBIOLOGY LABORATORY THURSDAY @

11:00 IN TRAKL HALL SESSION CHAIR: ED MOORE - UNIVERSITY OF GOTHENBURG .................................................... 68 • SESSION 5 • TRACK 6 • PRACTICAL TRAINING: INTERNAL STANDARDS THURSDAY @ 11:00 IN DOPPLER HALL

SESSION CHAIR: RUSS GRANT - LAB CORP ........................................................................................................... 69 • SESSION 6 • TRACK 1 • KEYNOTE FOR METABOLOMICS THURSDAY @ 14:30 IN MOZART 1-3 SESSION CHAIR: RUSS GRANT - LAB CORP ................................................................................................................................. 70 • SESSION 6 • TRACK 2 • MONITORING METABOLITES IN INBORN ERRORS OF METABOLISM THURSDAY @

14:30 IN MOZART 4-5 SESSION CHAIR: ZDENEK SPACIL - MASARYK UNIVERSITY ....................................................... 71 • SESSION 6 • TRACK 3 • ONCOLOGY THURSDAY @ 14:30 IN PAPAGENO HALL SESSION CHAIR: TBA (EXPECTED

TO BE YURI VAN DER BURGT, PENDING ACCEPTANCE OF OFFER) ............................................................................... 72 • SESSION 6 • TRACK 4 • TISSUE IMAGING KEYNOTE THURSDAY @ 14:30 IN PARACELSUS HALL SESSION CHAIR: KRISTINA SCHWAMBORN - TUM ....................................................................................................................... 73 • SESSION 6 • TRACK 5 • BIOMARKERS IN INFECTIOUS DISEASE THURSDAY @ 14:30 IN TRAKL HALL SESSION

CHAIR: STEFAN ZIMMERMAN ............................................................................................................................ 74 • SESSION 6 • TRACK 6 • PRACTICAL TRAINING : GETTING TO LOWER LIMITS OF QUANTITATION THURSDAY @

14:30 IN DOPPLER HALL SESSION CHAIR: BRIAN RAPPOLD - LAB CORP .................................................................... 75 POSTER PRESENTATIONS ..................................................................................................................................... 77

ENDOCRINOLOGY : POSTER PRESENTATIONS ....................................................................................................... 78 GLYCOMICS : POSTER PRESENTATIONS .............................................................................................................. 81 METABOLOMICS : POSTER PRESENTATIONS ........................................................................................................ 81 MICROBIOLOGY : POSTER PRESENTATIONS ......................................................................................................... 86 PROTEOMICS : POSTER PRESENTATIONS ............................................................................................................ 86 SMALL MOLECULES : POSTER PRESENTATIONS .................................................................................................... 89 TISSUE IMAGING : POSTER PRESENTATIONS ........................................................................................................ 97 TROUBLESHOOTING : POSTER PRESENTATIONS .................................................................................................... 98 VARIOUS OTHER : POSTER PRESENTATIONS ........................................................................................................ 99

PRESENTER INDEX ............................................................................................................................................. 101 MAP: 1ST FLOOR - EXHIBIT HALL ......................................................................................................................... 103 MAP: TRACKS AND FLOORS ............................................................................................................................... 104

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Sponsorship & Travel Grant Support

Gold

$12,

300

Silv

er

$6,8

00

Trav

el G

rant

Sup

port

$8,0

00

$2,0

00

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Scientific Committee

Please take a moment to acknowledge the members of the Scientific Committee who are pivotal in development of the Scientific Program.

Jean Armengaud, PhD CEA-Marcoule, France :: Microbiology

Christa Cobbaert, PhD Leiden University Medical Center :: Proteomics

Warrick Dunn, PhD University of Birmingham :: Metabolomics

Graeme Eisenhofer, PhD University of Dresden, Germany :: Endocrinology

Flaminia Fanelli, PhD University of Bologna - S.Orsola-Malpighi General Hospital, Italy :: Endocrinology

Tom Fiers, MDUniversity of Ghent, Belgium :: Endocrinology

Roland Geyer, PhD Thermo Fisher Diagnostics :: Practical Training

Ron Heeren, Prof. Dr. Maastricht University :: Tissue Imaging

David Herold, PhD, MD UCSD / VA Medical Center, San Diego :: Chair

Éva Hunyadi-Gulyás, PhD Biological Research Centre :: Proteomics

Julijana Ivanisevic, PhD University of Lausanne :: Metabolomics

Brian Keevil, PhDUniversity Hospital of South Manchester, Manchester, UK :: Endocrinology

Ido Kema, Prof. Dr. University Medical Center Groningen, The Netherlands :: Small Molecules

Gordan Lauc, PhD University of Zagreb; Genos :: Glycomics

Dirk Lefeber, PhD Radboudumc :: Glycomics

Sylvain Lehmann, MD, PhD University Hospital Center of Montpellier:: Proteomics

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Oleg Mayboroda, PhD Leiden University Medical Center, The Netherlands :: Metabolomics

Edward Moore, PhD University of Gothenberg, Sweden :: Microbiology

Laura Owen, PhD University Hospital of South Manchester :: Endocrinology :: Practical Training

Tiffany Porta, PhD Maastricht MultiModal Molecular Imaging (M4I) institute :: Tissue Imaging

Lars Melholt Rasmussen, MD, DMSc Odense University Hospital, University of Southern Denmark :: Proteomics

Kristina Schwamborn, MD, PhD Institute of Pathology, TU Munich :: Tissue Imaging

Judy Stone, PhD Center for Advanced Laboratory Medicine (CALM) at the University of California, San Diego :: Practical Training

Raf Van de Plas, PhD Delft University of Technology :: Tissue Imaging

Jody van den Ouweland, PhDCanisius-Wilhelmina Hospital, The Netherlands :: Small Molecules

Grace van der Gugten Provincial Health Services Authority, Canada :: Practical Training

Michael Vogeser, Prof. Dr. med.Institute of Laboratory Medicine, Hospital of the University of Munich, Germany for the working group LC-MS/MS of the DGKL :: Small Molecules

Elizabeth Want, PhD Imperial College London :: Metabolomics

Michael Wright LGC :: Practical Training

Manfred Wuhrer, Prof. dr. LUMC :: Glycomics

Stefan Zimmerman, MD University Hospital Heidelberg, Germany:: Microbiology

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General Information Smoking Smoking is prohibited within the congress facility. If smoking outside the congress center please refrain smoke within 15m of any doorway. Congress Badges Your badge constitutes your admission pass to the Congress Center. Please display your badge prominently while attending the congress and all associated functions. If you have not registered you will be escorted to the registration desk to get one, or off the premises. If you determined to be sharing badges, your badge will be confiscated and you will be escorted off the premises. Parking Sheraton Garage - entrance via Auerspergstraße with direct access to Salzburg Congress. Mirabell-Kongress-Garage - entrance via Mirabellplatz, 2-3 minutes walk to Salzburg Congress. Breakfast It is recommended that you take breakfast before arriving at the Congress Center, although each morning we will have a welcome coffee with a limited selection of light baked goods. Tape Recording/Video Recording Policy Please observe the MSACL policy which prohibits operation of tape recorders, video recorders, cameras, or camera phones, except for official association equipment, at all congress sessions, committee meetings, in the Exhibit Hall, and during the plenary sessions. If you want to see a copy of a poster, check to see if it is already uploaded online. Note: Throughout Conferences MSACL may be videotaping and taking photographs to be used for promotional or educational purposes by MSACL. If you do not wish to appear on camera, please notify the videographer or photographer and your request will be honored, or contact us after the congress to share your interest in not appearing in any MSACL promotional material.

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Presenter Info and Guidelines Podium Presentations Locations: Mozart Hall, Papageno Hall, Paracelsus Hall, Trakl Hall, Doppler Hall • If an individual is unable to present or does not show, the presentation time slot will be left open. IT

WILL NOT BE FILLED BY THE NEXT SPEAKER. The next speaker will begin presenting at his/her scheduled time.

o Back-Up Presenters: If a presenter does not show a back-up presenter may be called to fill in the open spot. Session Chairs, please contact registration immediately on determining that a speaker may not show so that efforts may be put in place to locate a back-up speaker

• Speakers: Please make an effort to repeat any questions from the audience before answering. • Podium presentations are 20 minutes including Q&A. • PC Laptops running Windows 7 Enterprise & Office 2010 will be provided. • Presenters should check-in 15-20 minutes prior to their Session (NOT their talk) with either the

Session Chair or AV Support on-hand to upload their presentation files to the primary presentation lap-top computer.

• Presenters should bring their presentations on thumb (USB) drives for placement on a single presentation computer from which all presenters will access their PowerPoint presentations.

• Laser pointers will be provided.

Poster Presentations Location: 1st Floor/ Exhibit Hall Posters will be placed for the entire conference starting on Tuesday at 17:30. Posters will be up for the duration of the congress. Please see poster abstract information for attendance details. • Poster dimensions should be A0 (841 x 1189 mm) in PORTRAIT format. • Poster Boards are plastic. • Adhesive velcro to place your poster WILL BE provided.

MONDAY

11:0019:00

11:0014:00

13:0014:00

18:0018:30

18:0018:30

18:00

18:00

18:3021:00

LUNCH ON YOUR OWNYour Choice

BADGE PICKUPEntrance Foyer

LUNCHEntrance Foyer

PETITE RECEPTIONEntrance Foyer

PETITE RECEPTIONEntrance Foyer

DINNER ON YOUR OWNYour Choice

DINNER ON YOUR OWNYour Choice

PRIVATE: SciCom Discussion GroupTBA

14:0018:00

09:0013:00

14:0018:00

SHORT COURSESMozart, Papageno, Paracelsus, Trakl, Doppler



08:0009:00

WELCOME COFFEEEntrance Foyer

COFFEE BREAKSEntrance Foyer

EVERY:50 MINS

FOR:10 MINS


EVERY:50 MINS

FOR:10 MINS

SUNDAY TUESDAY

08:0016:00

13:4514:00

14:0014:45

17:30

15:0016:00

13:3013:45

PLACE POSTERS1st Floor Exhibit Hall

CMS JOURNAL OVERVIEWEuropa Hall

OPENING PLENARY LECTUREEuropa Hall

EXHIBITS OPEN1st Floor Exhibit Hall

18:0019:00

MEET-THE-EXPERTS: Booth Tours1st Floor Exhibit Hall

STATE OF THE SCIENCE ADDRESSEuropa Hall

WELCOME, INTRODUCTION, & ORIENTATIONEuropa Hall


08:0009:00

09:0012:30



COFFEE BREAKEuropa Foyer

COFFEE BREAKEuropa Foyer

EXHIBITOR RECEPTION1st Floor Exhibit Hall

14:4515:00

16:0016:30

17:3020:00

22:00 ENJOY THE CITYYour Choice

EVERY:50 MINS

FOR:10 MINS

12:3014:00

12:30

OPENING LUNCHEuropa Hall

CONGRESS OPEN

19:0020:00

TROUBLESHOOTING POSTER ROUNDS : Part I1st Floor Exhibit Hall

20:0022:00

DISCUSSION GROUP - SWOT: The road to Comprehensive MS Implementation in the ClinicMozart 4-5

16:3017:00

POSTER LIGHTNING TALKSEuropa Hall

17:0017:30

EXHIBITOR LIGHTNING TALKSEuropa Hall

WEDNESDAY THURSDAY

08:0008:45

16:3017:30

19:3020:30

11:0012:00

14:3015:30

09:0010:00

CORPORATE WORKSHOPSPapageno, Paracelsus

DISTINGUISHED CONTRIBUTION AWARD

PLENARY LECTUREMozart 1-5

DISCUSSION GROUPSMozart 4-5, Papageno

SCIENTIFIC SESSION 2 Mozart, Papageno, Paracelsus, Trakl, Doppler

SCIENTIFIC SESSION 3 Mozart, Papageno, Paracelsus, Trakl, Doppler

SCIENTIFIC SESSION 1Mozart, Papageno, Paracelsus, Trakl, Doppler


07:4509:00

INTERMISSIONEntrance Foyer


EXHIBITOR RECEPTION1st Floor Exhibit Hall

14:1514:30

17:3019:30

20:30 ENJOY THE CITYSalzburg Old City

08:4509:00

12:0013:30

LUNCHExhibit Hall on 1st Floor

13:3014:15

CORPORATE WORKSHOPSMozart, Paracelsus, Trakl

10:0011:00

MEET-THE-EXPERTS: Poster Tours1st Floor Exhibit Hall

10:0011:00

POSTER SESSION 11st Floor Exhibit Hall

TROUBLESHOOTING POSTER ROUNDS : Part II1st Floor Exhibit Hall

15:3016:30

POSTER SESSION 21st Floor Exhibit Hall

15:3016:30


18:3019:30

MEET-THE-EXPERTS: Office Hours Forum1st Floor Exhibit Hall

08:0008:45

09:0010:00

11:0012:00

09:0010:00

CORPORATE WORKSHOPParacelsus

SCIENTIFIC SESSION 4Mozart , Papageno, Paracelsus, Trakl, Doppler


EXHIBITOR FEEDBACK SESSION1st Floor Cafe


07:4509:00

POSTER CONTEST: Finalist InterviewsExhibit Hall


AFTER CONGRESS MEET-UPAugustiner Brauhaus

CLOSING PLENARY LECTUREMozart 4-5

CLOSING RECEPTION & DISCUSSIONEntrance Foyer

08:4509:00

18:0022:00

16:0016:45

16:4518:00

10:0011:00

12:0013:30

LUNCH1st Floor Exhibit Hall

15:4516:00

14:00

POSTER AWARD PRESENTATION Mozart 4-5

Exhibits ClosEd

08:0009:00

SCICOM DISCUSSION GROUP1st Floor Cafe

10:0011:00


12:3013:30


13:3014:00

CORPORATE WORKSHOPPapageno

14:1515:00


14:3015:30


RECEPTIONMozart 4-5

15:3015:45

10:0011:00

POSTERS SESSION 31st Floor Exhibit Hall

12:3013:30

POSTERS SESSION 41st Floor Exhibit Hall

12:0013:30

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Plenary & Keynote Speaker Series Distinguished Contribution Award Plenary Lecture

>> Wednesday 16:30 in Mozart 1-5Mass Spectrometry and the Elucidation of Steroid Metabolome Cedric Shackleton UCSF Benioff Children’s Hospital Oakland Research Institute, Oakland, California, and Institute of Metabolism and Systems Research (IMSR), College of Medical and Dental Sciences, University of Birmingham, UK Sterols and steroids were among the first biomolecule groups to be studied by mass-spectrometry. The early success of GC separation in 1960 by Sweeley and Horning and the combination of GC and MS by Ragner Ryhage in Stockholm with invention of a technique for separating the bulk of carrier gas from analytes prior to ionization were fundamental. This led in the mid-60s to the first commercial dedicated GC/MS instrument, the LKB 9000. Professor Jan Sjövall (a colleague of Dr Ryhage) in particular spearheaded studies of sterols and steroids leading to multiple publications of mass spectra, and steroid quantification of serum, urinary and biliary steroids. Developments in derivatization, capillary columns and computerization were all important constituents of these developments. This presenter (a student of Sjövall) was early involved in the clinical diagnostic use of GC/MS in the 1970s and has been involved in studying urinary steroids and defining new disorders through metabolic profiles ever since. Particle beam MS (eg. FAB) was introduced in the 80s and allowed for the first time analysis of underivatised steroids and steroid conjugates. Steroid LC/MS (Thermospray) arrived in 1987 but it was a decade later before ESI and LC/MS/MS made the use of MS in the routine steroid analytical lab a practicality. Now the technique has essentially replaced RIA in routine endocrinological applications. GC/MS is labour-intensive and was never really suited to individual serum hormone analysis but has always been powerful for profiling urinary steroid metabolites, which now forms a branch of the recently-termed metabolome discipline. Recently there has been a re-birth in urinary steroid analysis in diagnosis and several groups are working on transferring GC/MS methodologies to LC/MS/MS to avoid need of derivatization and shorten chromatography times. This is a challenge because many of the “metabolite-type” steroids have weak ESI ionization and poor CID sensitivity. Two steroid groups are particularly suited to LC/MS/MS and those are the steroid sulfates and glucuronides since they have an existing ionic center. Analysis of intact conjugates avoids the labour-intensive enzyme hydrolysis, and we a pursuing their analysis. A drawback here is the paucity of authentic reference materials. A huge challenge in steroid metabolomics is the presentation of diagnostic data in a form friendly to researchers and clinicians. Many recent publications have addressed this issue, including use of heat-maps and machine learning for metabolome diagnostic interpretation.

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Plenary Lectures

>> Thursday 16:00 in Mozart 4-5Mass Spectrometric Analysis of Glycomic Signatures of Cancer and Autoimmune Diseases: Towards Clinical Application Manfred Wuhrer LUMC Many major human diseases including various types of cancer and autoimmune diseases are associated with protein glycosylation changes. These changes are often instrumental in disease development and progression. In cancer, glycosylation changes of both tumor tissues and tumor-derived circulating antigens have been described. However, diagnostic test often merely rely on the measurement of antigen concentrations and fail to register glycosylation changes, largely due to the lack of suitable technology and workflows. In this presentation, examples will be given of recently developed mass spectrometric workflows to determine disease-associated glycosylation changes from tumor tissues and in the circulation, but also from other body fluids including urine and saliva. Promising markers will be presented including immunoglobulin G glycosylation, cancer glycoprotein antigens, as well as tissue glycosylation signatures revealed by mass spectrometry imaging. Steps towards the clinical translation of these markers will be discussed.

>> Tuesday 14:00 in EuropaMetabolomics Messages on Human Health and Disease Jurek Adamski Helmholtz Zentrum München The human metabolome represents functional read out of processes in health and disease. The metabolome is both stable and highly dynamic. Stable components are determined by genetics, genomic imprinting or physiological homeostasis. The dynamics originates from circadian rhythm, hormonal status, nutrition, environmental exposure, ageing, medication or disease. Despite its interlaced origin metabolome specifically reflects distinct processes. Metabolomics studies request therefore a special study design. Unique metabolomics signatures have been identified pre-disease (like type 2 diabetes), in disease progression (chronic kidney disease) or in companion diagnostics (drug action monitoring). Metabolomics informed diagnostics has great potential for selectivity, specificity and multiplexing of indications to be screened for.

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Keynote Lectures

>> Wednesday 9:00 in Track 3 (Papageno) : Session 1Clinical Proteomics: The Path Towards ImplementationHarald Mischak mosaiques therapeutics GmbH & University of Glasgow Clinical proteomics, the application of proteome analysis to clinical purpose, represents a major field in the area of proteome research. The aim of clinical proteomics is the improvement of clinical care based on (1) the identification and application of biomarkers, and (2) the suggestion of relevant therapeutic targets. These two areas have different requirements regarding specimens to be employed, technology, and data evaluation. While substantial efforts (especially in biomarker discovery, but also in the identification of therapeutic targets) are evident based on the large number of associated publications, only a few approaches have actually resulted in clinical application. In this presentation, key issues and major challenges in clinical proteomics will be discussed. Among these are: a) the definition of a clinical need and a context-of-use, b) selection of appropriate samples, sample preparation and analytical platform, c) application of appropriate statistics, d) demonstration of benefit in a well-powered clinical study and e) obtaining regulatory approval and reimbursement, to enable actual implementation. For several conditions and diseases, clinical proteomics has delivered solutions that are already being applied. Most successful developments are based on multi-marker panels that have demonstrated value in large studies (i.e., including at least several hundred patients). The results of these studies are expected to initiate a change in disease assessment: from diagnosis based on existing damage and therapy aiming to prevent deterioration, towards diagnosis based on molecular mechanisms, prior to observations of clinical symptoms, and therapy via correction of molecular anomalies, thereby preventing disease onset. This will also open a path towards personalized precision medicine, where intervention will be guided by molecular mechanisms, not morphological changes. It is becoming clear that the tools required to meaningfully apply clinical proteomics (i.e., potential biomarkers, relevant technology and bio-banked samples) are available. The move from discovery towards validation and application is not only urgently necessary, but within reach. Now, a change in objective, away from additional discovery studies and towards properly testing the plethora of potential biomarkers that have been described, is needed to demonstrate the practical value of clinical proteomics.

>> Wednesday 11:00 in Track 3 (Papageno) : Session 2Mass Spectrometry Harmonisation: Paediatric Steroid TalesRonda Greaves Murdoch Children’s Research Institute Our goal in laboratory medicine is to improve patient health by using laboratory tests as our tool to support medical decisions. The change from immunoassay to mass spectrometry (MS) analysis of steroids has significantly improved the specificity, accuracy and sensitivity of many steroids frequently analysed in children. However, MS assays are not infallible and recognised differences exist across the total testing process. There are demonstrated variations pre-analytically, analytically and post-analytically in methods and understanding what differences are critical and how results compare between laboratories is essential to close the gap between result differences between laboratories. In this presentation, we will explore the challenges and possible solutions for the harmonisation of paediatric steroid analysis by MS to support medical decisions.

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>> Wednesday 14:30 in Track 2 (Mozart 4-5) : Session 3Tackling Analytical Challenges in Sports Drug Testing by Mass Spectrometric ApproachesMario Thevis German Sport University, Cologne, Germany European Monitoring Center for Emerging Doping Agents (EuMoCEDA), Cologne/Bonn, Germany Sports drug testing laboratories are facing multifaceted challenges including the misuse of naturally/endogenously occurring substances, non-approved/discontinued drug candidates, urine manipulation, etc. In order to provide best-possible analytical performance, mass spectrometry-based approaches are predominantly utilized to detect prohibited substances and methods of doping. With the constantly increasing analytical requirements concerning the number of target compounds, the complexity and range of physico-chemical properties of analytes (e.g., inorganic ionic transition metals, gases, lipids, alkaloids, peptides, proteins, DNA/RNA-based drugs, etc.) as well as the desire to accelerate analyses and obtain information allowing also for retrospective data mining, high resolution/high accuracy mass spectrometry has become a mainstay in doping controls. In that context, various assays have been reported, enabling either multi-component analyses of low- or high molecular mass measurands or the specific and dedicated (confirmatory) detection of prohibited substances. Selected applications will be presented reporting on examples of recent findings in routine sports drug testing, demonstrating both the inventiveness of cheating individuals that undermine current anti-doping efforts as well as the relevance of in-depth investigations into unusual findings, where the athletes’ innocence was to be shown albeit prohibited substances were unequivocally identified in their doping control urine samples.

>> Wednesday 14:30 in Track 4 (Paracelsus) : Session 3Combining Mass Spectrometry Imaging & Microproteomics to Investigate Intratumor Heterogeneity Liam McDonnell Fondazione Pisana per la Scienza ONLUS, Pisa, Italy Mass spectrometry imaging (MSI) is able to simultaneously record the distributions of hundreds of molecules directly from tissue. This spatially-resolved molecular information can be combined with multivariate/clustering analysis to reveal regions of tissue with distinct molecular signatures, a process that has been termed MSI-based molecular histology and has been used to reveal tumor subpopulations, metabolically distinct cell layers, and tumor interface zones. Rapid direct tissue analysis is essential for MSI in order to maintain spatial localization and acceptable measurement times. The absence of an explicit analyte separation/purification step means MSI lacks the depth of coverage of LC-MS/MS. Here, we demonstrate how MSI can be combined with high sensitivity microproteomics, even of the same tissue section, to further investigate the molecular changes associated with tumor subpopulations.

>> Thursday 9:00 in Track 5 (Trakl) : Session 4Tandem MS-Proteotyping: Proteomics- and Genomics-Based Characterization and Typing of Infectious Bacteria Edward Moore University of Gothenburg, Sweden LC-MS/MS proteomics- and genomics-based characterisation and typing of microorganisms, i.e., ‘proteotyping’, using peptides from expressed proteins, matched to genomic sequence data, can be applied for sensitive and accurate detection and typing of pathogenic bacteria. Proteotyping is capable of resolving closely-related bacterial taxa with simultaneous detections of virulence and antibiotic resistance features, providing for comprehensive characterisations of infectious bacteria. The methodology may be applied directly to analyses of clinical samples without prior cultivation and isolation, thus providing for rapid, reliable, infectious disease diagnostics.

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>> Thursday 11:00 in Track 4 (Paracelsus) : Session 5High-throughput Glycomics in Patient Stratification - What Did We Learn from the First 60,000 Analyses Gordon Lauc University of Zagreb & Genos Glycoscience Research Laboratory, Zagreb, Croatia Since the onset of genome wide association studies, thousands of genetic loci have been associated with different diseases and traits. However, in the last few years it is becoming increasingly clear that variations in a DNA sequence are only a beginning of the understanding of complex human diseases. Genetic polymorphisms have to be put in the context of complex biology of life and a more elaborate approach that combines different ‘omics phenotypes is needed to understand disease mechanisms and perform patient stratification that transcends genomics. Glycomics, as by far the most complex epiproteomic modification, has an immense potential in this respect, which is only beginning to be investigated.

>> Thursday 14:30 in Track 1 (Mozart 1-3) : Session 6How Tandem Mass Spectrometry Revolutionized Newborn Screening David Millington Duke University - Retired Inspired by a clinician’s account of a child rescued from near death by a revolutionary therapeutic intervention, the author applied chemistry and mass spectrometry to solve an analytical challenge that led to the first front-line diagnostic test performed by tandem mass spectrometry (MSMS) – the analysis of acylcarnitines to recognize and diagnose inherited disorders of fatty acid and branched-chain amino acid catabolism. By applying this method to dried blood spots and adding an additional analytical component to include several essential amino acids, a novel multiplex assay was developed to screen newborns for over 30 inherited metabolic conditions with a single test. The introduction of this method into public health systems and hospitals across the world during the past 20 years has literally revolutionized neonatal screening, and new technology is being introduced to add even more value to this critical “first test”. This concept subsequently became the basis of targeted metabolomics platforms that have been used, for example, to help identify new animal models of metabolic disease by screening the offspring of genetically modified adults. MSMS with UPLC has been widely applied to develop new assays for useful biomarkers of metabolic disease for both diagnosis and therapeutic monitoring. Examples from the author’s laboratory will be used to illustrate the value and scope of these methods.

>> Thursday 14:30 in Track 4 (Paracelsus) : Session 6 Advances in Computational Methods for Tissue Imaging by Mass SpectrometryRaf Van De Plas Delft TU Imaging Mass Spectrometry (IMS) has made rapid progress as an imaging modality that can map the spatial distribution of molecules in tissue. In recent years, novel computational developments have become an increasingly important part of major advancements in this field. This talk presents several computational techniques developed in our group, specifically relevant to molecular imaging in medicine and the clinical practice. We show recent work in low-level signal processing, where in silico integration of isolation windows enables High-Dynamic-Range mass spectrometry, substantially increasing MS sensitivity. We also address advancements in data-driven image fusion, a multi-modal data mining methodology that drives the automated discovery of biomolecular relationships between stained microscopy and IMS, thus directly linking exploratory tissue analysis to established clinical targets.

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Young Investigator Grants Young Investigator Grants (n=78) are provided to support trainees (MD/residents/fellows and PhD - students / post-docs) and young faculty members (fewer than 4 years since appointment) who have submitted abstracts that have been accepted for presentation.

Mina Adam Imperial College London UK Khem Adhikari Rigshospitalet Laura Albantakis Max Planck Institute of Psychiatry Ida Bøgh Andersen University of Southern Denmark, Lillebaelt Hospital, Vejle Sandra Anjo Center for Neuroscience and Cell Biology – University of Coimbra (CNC-UC) Sebastiano Barco Istituto Giannina Gaslini Florian Barré M4I, Maastricht University Ilaria Belluomo Imperial College London Alexander Brzhozovskiy V. I. Kulakov National Medical Research Center for Obstetrics, Gynecology and Perinatology, Moscow, Russia Bo Burla Singapore Lipidomics Incubator (SLING), National University of Singapore Rory Cave University of East London Yasmine Chetouane IHU méditerranée infection Shosha Dekker Leiden University Medical Center Jeany Delafiori University of Campinas (UNICAMP) Lisa Delahaye Ugent Nina Denver University of Glasgow Neus Fabregat-Cabello University of Liège, CHU de Liège Jordi Farre-Segura University of Liège Claudia Fredolini Uppsala University Paulina Zofia Goryńska Nicolaus Copernicus University in Bydgoszcz, Poland Gonçalo Graça Imperial College London Nicola Gray University of Reading, UK Lucia Grenga CEA Antonina Gucciardi University of Padova Jörg Hanrieder University of Gothenburg Charlotte Harborow Liverpool Clinical Laboratories Emma Hurst University of Edinburgh Ólöf Gerdur Ísberg University of Iceland Karol Jaroch DNicolaus Copernicus University, Toruń, Poland Frédéric Jauffrit Université de Lyon Emine Kazanc Imperial College of London Moon-Ju Kim Yonsei University Natalia Kitsilovskaya National Research Center of Obstetrics and Gynecology, Russia Valeriia Kuzyk Vrije Universiteit Amsterdam Guinevere S.M. Lageveen-Kammeijer Leiden University Medical Center Marine Letertre Imperial College London Jaanus Liigand University of Tartu Craig Livie Glasgow Royal Infirmary Katarina Madunic Leiden University Medical Center Govinda Mandal Masaryk University, Czech Republic Pavel Markin Sechenov First Moscow State Medical University David Marshall Manchester University NHS Foundation Trust Elizabeth Mathew Manipal College of Pharmaceutical Sciences Melissa McNaughton University of Manchester Arsenty Melnikov Internatrional Tomography Center SB RAS Elham Memarian Leiden University Medical Centre Samuel Mesihää University of Helsinki, Department of Forensic Medicine Stephanie Mezger Maastricht University Marco Mezzullo University of Bologna Alan Moran Leiden University Medical Centre Mouton Nicolas University de Lyon Claude Bernard

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Ádám Pap University of Szeged, Hungary Isabella Piga University of Milano-Bicocca Martina Pirro Leiden University Medical Center LUMC Mark Pratt University Medical Centre Groningen Madlen Reinicke University Leipzig, Institute of Laboratory Medicine Dinara Salimova V.I. Kulakov National Medical Research Center of Obstetrics, Gynecology and Perinatology, Moscow, Russia Maria Laura Santoru University of Cagliari Thomas Senard Leiden University Medical Center Andrew Smith University of Milano-Bicocca Zdenek Spacil Masaryk University Natthida Sriboonvorakul Mahidol University Dominika Strzelecka University of Warsaw Eliška Stuchlíková Masaryk University Godwin Kwabena Tetteh Frimley Health NHS Trust Foundation Unnur Arna Thorsteinsdottir University of Iceland Robin Tiphaine CHU of Limoges/Shimadzu Corporation Federico Torta National University of Singapore Gábor Tóth MS Proteomics Research Group, Hungarian Academy of Sciences Vera van der Velpen University of Lausanne Tirsa van Duijl Leiden University Medical Center (LUMC) Ana Varela Coelho ITQB NOVA Martha Kampp Nøhr Vestergaard University of Iceland Patrick Wehrli University of Gothenburg Luke Whiley Imperial College London Georgia Woodfield Imperial College London Vadim Yanshole International Tomography Center SB RAS Lyudmila Yanshole International Tomography Center SB RAS

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Lab Director Grants

Lab Director Grants (n=4) are provided to individuals leading clinical labs. These individuals have had minimal exposure to mass spectrometry and are interested in gaining more understanding of its clinical applications.

Christiane Auray-Blais Université de Sherbrooke Sigrídur Klara Bödvarsdóttir BioMedical Center, University of Iceland Seán Costelloe Cork University Hospital Claude Hercend Beaujon Hospital

Lab Director & Trainee Travel Grants supported in part by:

Trainee Grants

Trainee Grants (n=19) are provided to individuals training to lead clinical labs. These individuals have had minimal exposure to mass spectrometry and are interested in gaining more understanding of its potential applications.

Julia Dittrich Leipzig University and University Hospital Leipzig Krzysztof Gorynski Nicolaus Copernicus University Collegium Medicum Kirsten Grant NHS Scotland Janine Grant Melbourne Health Malak Jaber University of Nottingham Susan Johnston NHS Greater Glasgow & Clyde Jennifer Lake Barts Health NHS Trust Louisa Lee NHS Greater Glasgow & Clyde Adrienn Molnar ELTE Eotvos Lorand University, Institute of Chemistry Wai Yan NG Toxicology Reference Laboratory, Hospital Authority, HK Sarah Pitkin Barts Health NHS Trust Dimitrios Platis Institute of Child Health Gitta Schlosser ELTE Eötvös Lorand University, Institute of Chemistry Andraz Smon University Medical Centre Ljubljana Jeroen van den Wijngaard Leiden University Medical Center Matthew Whitlock Northwest London Pathology Chen Xin BGI Research Institute Gao Xinyan BGI Research Institute Monowara Zaman Barts Health NHS Trust

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Short Course Overview Data Science 101 (Beginner to Intermediate) Breaking up with Excel: An Introduction to the R Statistical Programming Language Duration: Sunday Afternon → Tuesday at Lunch Location: Mozart 5 Instructor(s): Daniel Holmes, MD & William Slade, PhD

Data Science 201 (Intermediate) Going Further With R: Tackling Clinical Laboratory Data Manipulation and Modeling Duration: Sunday Afternon → Tuesday at Lunch Location: Mozart 4 Instructor(s): Patrick Mathias, MD, PhD & Randall Julian, PhD

Lab Medicine 101 (Beginner) Basics of Laboratory Medicine Duration: Monday Afternoon → Tuesday at Lunch Location: Mozart 3 Instructor(s): Prof. Dr. med. Michael Vogeser

LC-MSMS 101 (Beginner) Getting Started with Quantitative LC-MS/MS in the Diagnostic Laboratory Duration: Sunday Afternon → Tuesday at Lunch Location: Mozart 2 Instructor(s): Judy Stone, PhD & Grace van der Gugten

LC-MSMS 202 (Intermediate) Practical LC-MS Maintenance and Troubleshooting Duration: Sunday Afternon → Tuesday at Lunch Location: Paracelsus Hall Instructor(s): J. Will Thompson, PhD & Erik J. Soderblom, PhD

LC-MSMS 301 (Advanced) Development and Validation of Quantitative LC-MS/MS Assays for Use in Clinical Diagnostics Duration: Sunday Afternon → Tuesday at Lunch Location: Papageno Hall Instructor(s): Russell Grant, PhD & Brian Rappold

Metabolomics 202 (Beginner to Intermediate) Metabolomics: Approaches, Applications and Challenges Duration: Sunday Afternon → Tuesday at Lunch Location: Doppler Hall Instructor(s): Julijana Ivanisevic, PhD & Elizabeth Want, PhD

Proteomic Microbiology 201 (Beginner to Intermediate) Bottom-Up and Top-Down Proteomic Approaches for Bacterial Identification and Characterization, a Focus on MALDI-TOF and Advanced Technologies Duration: Monday Afternoon → Tuesday at Lunch Location: Trapp Zimmer Instructor(s): Jean Armengaud, PhD, Stefan Zimmermann, MD

Proteomics 201 (Intermediate to Advanced) Clinical Proteomics Duration: Sunday Afternon → Tuesday at Lunch Location: Trakl Hall Instructor(s): Cory Bystrom, PhD & Chris Shuford, PhD

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Discussion Groups

Sunday from 18:30 - 21:00 Scientific Committee Discussion Group TBA Lead: David Herold A meeting of the Scientific Committee to discuss the current congress. Dinner to be provided.

Tuesday from 20:00 - 22:00SWOT : The Road to Comprehensive MS Implementation in the Clinic Mozart 4-5 (Track 2) Lead: Daniel Holmes Panelists: Michael Vogeser, Hospital of the Univerity of Munich, Elizabeth Want, Imperial College London, Robert DeWitte, Thermo Scientific,Christa Cobbaert, Leiden University Medical Center The process of bringing mass spectrometry from esoteric research niche to high throughput clinical analyzer is more complex and nuanced than it initially appeared. Superior analytical technology is not all that is necessary to successfully cross the chasm from early adopters to early majority. A recent editorial by Zhang and Vogeser in Clinical Mass Spectrometry, Understanding the strategic landscape surrounding the implementation of mass spectrometry in the clinical laboratory: A SWOT analysis (Open Access) provides an overview of the current landscape by highlighting the strengths, weaknesses, opportunities and threats that support and temper the path towards wide-spread implementation. This discussion group is intended to bring this reality under a brighter light to increase awareness and, thereby, stimulate discussion, collaboration and strategizing.

Wednesday from 19:30 - 21:00Development of LC-MS Certification Programs Mozart 4-5 (Track 2) Lead: Judy Stone Panelists: Fumio Nomura, Chima University Hospital, Ronda Greaves, Murdoch Children's Research Institute. Michael Vogeser, University Hospital, LMU Munich. Daniel Holmes, St Paul's, Vancouver, Canada, Joshua Hayden, Weill Cornell Medicine One major barrier to implementation of mass spectrometry (MS) in diagnostic laboratories is the lack of staff trained for use of the technique in a production setting. A variety of training options exist, but for most laboratories the assumption is that staff will have to be trained onsite. We queried the MSACL community about clinical MS training options and present a summary of the responses at https://www.msacl.org/certification_program_survey.php. This discussion session will explore the question of certification (why, who, how) as an aid (or perhaps a barrier) to increasing the availability and recognition of competency for users of clinical mass spectrometry.

Design of Experiments – get it right from the beginning Papageno Hall (Track 3) Lead(s): Margrét and Unnur Thorsteinsdóttir We will demonstrate how method development can become much more efficient by utilizing design of experiments (DoE). DoE offers a practical approach for performing experiments in accordance to predefined plan, modelling by empirical functions, and graphical visualization. Example from optimization of a UPLC-MS/MS method for clinical application will be used to show the cost-effective benefit of DoE, where it allows the effect of variables to be assessed with only a fraction of the experiments that would be required by changing one-separate-factor-at-time (COST) approach and spots critical setting early on.

Thursday from 8:00 - 9:00Scientific Committee Discussion Group 1st Floor Cafe Lead: David Herold A meeting of the Scientific Committee to review the outcome current congress and plan for future events.

Thursday from 9:00 - 10:00Exhibitor Feedback Group 1st Floor Cafe Lead: Chris Herold An opportunity for exhibiting vendors to provide feedback on what worked and what could use improvement.

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Exhibits Summary

Tuesday 8:00 – 16:30 Exhibitor Set-Up (EXHIBITS CLOSED) – Poster Placement for Presenters Permitted.

17:30 – 20:00 Opening Reception in Exhibit Hall Wednesday

10:00 - 11:00 Coffee Break in Exhibit Hall 12:00 – 13:30 Lunch provided in Exhibit Hall . 15:30 – 16:30 Coffee Break in Exhibit Hall 17305 – 19:30 Reception in Exhibit Hall

Thursday 9:00 – 10:00 Exhibitor Feedback Session in LowerCafe, 1st Floor 10:00 - 11:00 Coffee Break in Exhibit Hall12:00 – 13:30 Closing Lunch in the Exhibit Hall.

13:30 EXHIBITS CLOSED13:30 – 16:45 Exhibitor Breakdown and Packing in Exhibit Hall Only

16:45 Package Pickup from Loading Bay Allowed to Begin 21:00 Deadline for removal of Exhibits from Exhibit Hall

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Exhibitors Agilent Technologies Booth #19-20 http://www.agilent.com With an industry leading portfolio of analytical products for your clinical research laboratory, Agilent Technologies delivers everything your laboratory needs from sample preparation to final answers. From automation and sample preparation technologies, columns and consumables, laboratory informatics, to liquid and gas chromatography systems, and mass spectrometry systems such as ICP-MS, GC/MS, and LC/MS, Agilent provides premiere analytical solutions that will ensure confident identification and quantitation of both endogenous and exogenous substances in complex biological matrices with the utmost accuracy, productivity and reliability.

Biocrates Life Sciences Booth #6 http://www.biocrates.com Biocrates and Metanomics Health have recently merged to create a new global leader in Metabolomics and early disease detection. As a total solution provider, Biocrates and Metanomics Health provide the broadest technology and product portfolio in the industry. The merged companies have extensive expertise in targeted and global metabolic profiling, customized assay development, targeted screening kits, and data interpretation. Finally, capabilities in CDx / Diagnostic Kit development make Biocrates and Metanomics Health the preferred partner for all your requirements in metabolite analysis. Biocrates’ metabolomics kits allow for the analysis of up to 400 metabolites across multiple analyte classes. These kits have contributed to more than 800 scientific publications and are being used in >100 mass spectrometry laboratories throughout the world. The whole range of analytical and data interpretation services is available through the company’s service laboratories in Berlin, Germany and Innsbruck, Austria.

Cambridge Isotope Labs Booth #8 http://www.isotope.com Cambridge Isotope Laboratories, Inc. is the world leader in the manufacture and separation of stable isotopes and isotope-labeled compounds. CIL and Euriso-Top (a European subsidiary of CIL) offer highly pure compounds that are uniformly or selectively enriched in 13C, 15N, D, 18O or 17O. CIL’s labeled reagents are used in proteomics, metabolomics, metabolism, and environmental applications for quantitative mass spectrometry. Our products include MRM PeptiQuantTM assay kits, SILAC protein quantitation kits, media and reagents, 99% enriched amino acids, Mouse Express® Lys 13C6 and 15N mouse feed and tissue, 15N spirulina, intact labeled proteins, growth media for protein expression, cell-free protein synthesis products, environmental contaminants standards for ultra-trace analysis, steroids, acylcarnitines, drug metabolites, nucleic acids, lipids and carbohydrates. CIL has cGMP capabilities; a majority of substrates can be manufactured to Q7A compliance.

Chromsystems Booth #12-13 http://www.chromsystems.com Chromsystems is a leading global company providing ready-to-use kits, multilevel calibrators and quality controls for routine clinical diagnostics by LC-MS/MS and HPLC. Our parameter menu covers a range of areas such as newborn screening, therapeutic drug monitoring, steroid analysis, vitamin profiling and more. We continuously expand our portfolio with additional tests all ensuring a highly accurate and cost-effective analysis. We enable laboratories to add new parameters into their diagnostic routine and expand their testing menu without prior technical expertise. They can immediately start the analysis with a minimum of time for the sample preparation. The products are comprehensively validated, and in particular LC-MS/MS methods with all widely used tandem mass spectrometers. They are CE-IVD compliant, satisfying regulatory requirements in the laboratory. We combine these high quality products with an excellent support programme and service for our customers.

Immundiagnostik Booth #11 http://www.immundiagnostik.com Immundiagnostik AG is a globally operating diagnostics company represented in over 30 countries. We focus on the development and production of innovative immunoassays (ELISA, EIA) and other analytical detection methods (e.g. HPLC, LC-MS/MS and PCR) for medical routine and research. Our mission is to provide effective tools for prevention, differential diagnosis and therapy monitoring in the areas of gastroenterology, cardiovascular diseases, disorders of the skeletal system and oxidative stress. The product portfolio is completed by a broad range of antibodies and antigens. Our business relations include contract analyses for diagnostic laboratories and academic research institutions, esp. in context with clinical trials. Securing progress: Our comprehensive range of products is continuously refuelled by a rich pipeline of proprietary developments. With a headcount of more than 80 employees, Immundiagnostik’s headquarter is located in Bensheim, south-western Germany.

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Merck Booth #4 http://www.sigmaaldrich.com/clinical The life science business of Merck combines strong innovative R&D teams with a global network spanning more than 60 countries and 70 manufacturing sites. Innovations include BioSPME for rapid extraction from biological matrices for LC/MS or direct MS, in-line SPE cartridges for effective extraction and phospholipid removal and the latest developments in monolith and fused core HPLC column technologies, all of which can be seen on booth 4. The company provides a product portfolio of 300,000 products, including over 20,000 reference materials, along with Cerilliant Certified Reference Materials, all with easy access to comprehensive data through one of the most advanced web platforms.

PerkinElmer Booth #25 http://www.perkinelmer.com PerkinElmer is the global market leader in neonatal screening, currently serving customers in more than 100 countries. We are a total solution provider offering complete systems based on a broad range of high quality, validated products, including newborn screening kits, consumables, instruments and software. PerkinElmer is proud to offer the QSight 220 for Clinical Research System for use in Clinical Research environments that require sensitivity, robustness and reduced downtime. Come visit us at booth 25 to learn more!

PROMISE Advanced Proteomics Booth #18 https://promise-proteomics.com/ Promise Proteomics offers a range of “off the shelf” and custom produced stable isotope labelled intact, recombinant proteins to benefit scientists who require robust and reliable quantitative LC-MS workflows Our labelled proteins are designed to benefit these areas: • Biomarkers evaluation and validation: Labelled proteins are the gold standard to achieve reproducible, reliable and accurate data. • Pharmacokinetics studies of biotherapeutics: Promise produces custom labelled proteins for PK studies. Additionally, we provide services for monoclonal antibody development in collaborative R&D projects with pharmaceutical partners • Development of Certified Reference Materials: We produce both labelled and labelled proteins tailored to customer specifications, for example purity and quantity. • Therapeutic Drug Monitoring: Promise offers labelled antibodies and sample processing kits for monitoring patients treated with biotherapies. Visit our website www.promise-proteomics.com to learn more about our catalogue of labelled proteins and custom production services

RECIPE Chemicals + Instruments Booth #17 http://www.recipe.de/en/index.html Starting business in 1982, RECIPE is one of the leading companies in HPLC and LC-MS/MS diagnostics today. For mass spectrometry, RECIPE offers CE/IVD labelled ClinMass® LC-MS/MS Complete Kits. Furthermore, several reagents such as ClinMass® Optimisation Mixes and Internal Standards, ClinCal® Calibrators and ClinChek® Controls are available for a reliable and standardised LC-MS/MS analysis. All products are developed and produced in our state-of-the-art production plant in Munich. RECIPE is recognised worldwide as a reliable partner for clinical laboratories and is certified by the quality management standards EN ISO 9001 and 13485.

Restek Booth #22 http://www.restek.com A leading innovator of chromatography solutions for both LC and GC, Restek has been developing and manufacturing columns, reference standards, sample preparation materials, accessories, and more since 1985. We provide analysts around the world with products and services to monitor the quality of air, water, soil, food, pharmaceuticals, chemicals, and petroleum products. Our experts have diverse areas of specialization in chemistry, chromatography, engineering, and related fields as well as close relationships with government agencies, international regulators, academia, and instrument manufacturers. www.restek.com

SCIEX Booth #29-30 http://www.sciex.com SCIEX’s global leadership and world-class service and support in the capillary electrophoresis and liquid chromatography-mass spectrometry industry have made it a trusted partner to thousands of the scientists and lab analysts worldwide who are focused on basic research, drug discovery and development, food and environmental testing, forensics and clinical research. As part of AB SCIEX, SCIEX Diagnostics brings the power, flexibility, reliability, and accuracy of mass spectrometry technology to clinical testing laboratories. Offering an expanding portfolio of mass spectrometry based solutions and assays for in vitro diagnostic use, SCIEX Diagnostics enables customers to deliver high quality diagnostic information to clinicians who make decisions affecting patient care.

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Shimadzu Booth #9-10 https://www.shimadzu.eu Shimadzu is one of the worldwide leading manufacturers of analytical instrumentation. Its equipment and systems are used as essential tools in all areas of clinical research. Since more than 140 years, Shimadzu is at the service of science ensuring precise and reliable analyses. Among the leaders in GCMS as well as in LCMS, Shimadzu is offering a full range of LC-MS/MS systems to fit all needs depending on the application, and will introduce new solutions for screening and quantitation in toxicology field. In addition, the unique and flexible fully automated sample preparation system seamlessly integrated with our LC-MS/MS range will be shown: CLAM-2000. Take the opportunity to visit our booth “9 & 10”.

Tecan Booth #27-28 http://www.tecan.com Tecan (www.tecan.com) is a leading global provider of laboratory instruments and solutions in biopharmaceuticals, forensics and clinical diagnostics. The company specializes in the development, production and distribution of automated workflow solutions for laboratories in the life sciences sector. Its clients include pharmaceutical and biotechnology companies, university research departments, forensic and diagnostic laboratories. As an original equipment manufacturer (OEM), Tecan is also a leader in developing and manufacturing OEM instruments and components that are then distributed by partner companies. Founded in Switzerland in 1980, the company has manufacturing, research and development sites in both Europe and North America and maintains a sales and service network in 52 countries. In 2016, Tecan generated sales of CHF 506 million (USD 511 million; EUR 464 million). Registered shares of Tecan Group are traded on the SIX Swiss Exchange (TECN; ISIN CH0012100191).

Thermo Scientific Booth #14-16 http://www.thermoscientific.com/msacleu Thermo Fisher Scientific Inc. is the world leader in serving science, with revenues of $17 billion and approximately 50,000 employees in 50 countries. Our mission is to enable our customers to make the world healthier, cleaner and safer. We help our customers accelerate life sciences research, solve complex analytical challenges, improve patient diagnostics and increase laboratory productivity. Through our premier brands – Thermo Scientific, Applied Biosystems, Invitrogen, Fisher Scientific and Unity Lab Services – we offer an unmatched combination of innovative technologies, purchasing convenience and comprehensive support. For more information, please visit www.thermofisher.com.

UTAK Laboratories Booth #1 http://www.utak.com At UTAK, labs get the control they need for their clinical and forensic toxicology test methods. We craft quality controls for every kind of analysis, including comprehensive stock controls in urine, serum, blood, oral fluid; starting matrices for labs developing in-house QC material; and personalized control solutions to support new lab-developed methods. Better control, more accurate results and improved safeguarding of health and safety standards—it’s why we call ourselves “control freaks.”

Waters Booth #23-24 http://www.waters.com/waters/eventInstance.htm?eiid=134989704 Waters Corporation, the premium brand in the analytical instruments industry, creates business advantages for laboratory-dependent organizations by delivering practical and sustainable scientific innovation to enable significant advancements in healthcare delivery, environmental management, food safety, and water quality worldwide. Bringing keen understanding and deep experience to those responsible for laboratory infrastructure and performance, Waters helps customers make profound discoveries, optimize laboratory operations, deliver product performance, and ensure regulatory compliance. Pioneering a connected portfolio of separations and analytical science, laboratory informatics, mass spectrometry, as well as thermal analysis, Waters’ technology breakthroughs and laboratory solutions provide an enduring platform for customer success.

Zivak Technologies Booth #21 http://www.zivak.com Based in Istanbul, Turkey, Zivak Technologies provides ready to use LC-MS/MS and HPLC analysis kits in the clinical diagnostic field. The company also offers its own fully automated sample preparation and injection system which enables laboratories around the globe to make efficient use of their LC-MS/MS instruments as well as HPLC instruments in a fast, accurate and cost efficient way.The company focuses on automation of processes in labs, HPLC and LC-MS/MS analysis kits, and consumabled. Sales and marketing activities are carried out in more than 70 countries.

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Corporate Workshops

Wednesday

08:00 & 13:30

Thursday 08:00 & 13:30

Mozart 1-3 Ground Floor Mozart 4-5 Ground Floor Papageno Hall Ground Floor Paracelsus Hall 2nd Floor Trakl Hall 3nd Floor

If you have an MSACL registration badge you may attend any Corporate Workshop.

Sponsoring vendors may request that attendees register, but it is not required. Vendors may, however, provide priority seating to pre-registered workshop attendees

if there are space limitation issues.

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Corporate Workshops - Wednesday

8:00 - 8:45 Thermo Scientific - Papageno Hall Targeted screening and semi-quantitation of drugs in plasma using high-resolution accurate-mass detection and online sample preparation Sensitive and selective analytical methods are required in clinical research and forensic toxicology to properly identify a broad range of analytes in complex matrices. The existing methods need to be constantly updated as new compounds, such as designer drugs, become readily available to the illicit drug market. Liquid chromatography coupled to mass spectrometry (LC-MS) has been widely used in this area for years. So far, the preferred detection technology has been triple quadrupole MS for its selectivity in selected reaction monitoring (SRM) mode. However, this approach suffers from the limitation of being able to perform only targeted analysis, while not useful for unknown screening of new drugs. Come to our educational seminar to learn more. For Research Use Only. Not for use in diagnostic procedures.

SCIEX - Paracelsus Hall A Clinical Method for Determination of Anti-Infectious Drugs in Serum Anders Blomgren Associate professor at Division of Clinical Chemistry and Pharmacology, Lund University The initiative for this work was taken by physicians from four different disciplines; intensive care, clinics for infection, thorax and transplantation. There has been an unmet need for these types of analysis and especially with short response times. A TDM wish list was produced that containing about 25 different anti-infection drugs and we agreed on starting to develop methods for 7 of them as a first step. During this talk the following will be presented. • Introduction • Analytes – chemistry/properties/stability • LC, MS and Sample prep development and problem solving • Validation with results • Conclusion and future

Corporate Workshops - Wednesday

13:30 - 14:15 Shimadzu - Mozart 1-3 Better Ways of Working in Routine Clinical Pathology Laboratories Healthcare is dynamic and knows no bounds and so routine clinical pathology laboratories face increasing pressures on cost, time and delivering actionable data. At Shimadzu, we believe there's always a way to make life better. This seminar explores a few options to make a difference. 1) Neil LOFTUS - SHIMADZU Corporation, UK Expanding the capability of mass spectrometry in routine clinical pathology – creating better ways of delivering actionable data. 2) Andrea Radeljak, Matea Ivić – KB Merkur, Croatia Automating standardized workflows in the clinical laboratory – using the Shimadzu CLAM-2000 integrated with LC-MS/MS to help increase sample throughput and make it easier for users. 3) Sigrid BAUMGARTEN - Alsachim SAS, France From core expertise of Stable Isotope Labelled Compounds to reagent kits for high quality LCMS data Disclaimer: For Research Use Only. Not for use in diagnostic procedures. Not available in the USA, Canada and China.

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Thermo Scientific - Mozart 4-5 Fully Automated LC-MS – Theory becomes Reality The use of LC-MS in diagnostic pathology has been limited by the complexity of equipment, materials and skills required to establish a fully validated service. A review showed only 19% of respondents were running VitaminD using LC-MS. On the basis that this is the gold standard, a fully validated and simple-to-use mass spectrometer has been a long awaited development. This presentation shares the first practical experience of using the Thermo Scientific™ Cascadion™ SM Clinical Analyser, in a laboratory, where staff have no prior experience of using LC-MS technology. Automated LC-MS/MS: A disruptive technology will improve the quality of care for our patients The LC-MS/MS platform provides solutions to complex analytical problems for clinical routine laboratories. This development started decades ago and the technology has proven to deliver reliable results with unreached specificity in TDM, DoA testing, endocrine diagnostics and newborn screening. In this presentation the concept of a fully integrated and automated LC-MS/MS is described. Special focus is on features of the Thermo Scientific™ Cascadion™ SM Clinical Analyzer that can overcome the limitations of open systems. Product IVD/CE-marked. Product not 510(k) cleared and not yet available for sale in the US.

Agilent Technologies - Paracelsus Hall Developing and Delivering Advanced Protein Biomarker Tests for an Era of Precision Medicine Speaker: Prof. Stephen R Pennington, Senior Fellow UCD Conway Institute, University College Dublin, Ireland This presentation will address the key issues around the development of protein signatures in areas of significant unmet clinical need and potential strategies for the delivery and implementation of multiplexed protein biomarkers in a diagnostic setting. View abstract at www.agilent.com/en/promotions/msacleu2018 For Research Use Only. Not for use in diagnostic procedures.

Waters - Trakl Hall Determination of Steroid Hormones in biological matrices by Tandem Mass Spectrometry The determination of steroid hormones in biological matrices requires high analytical sensitivity and is particularly challenging due to analytical interferences from structurally related steroid hormones and synthetic derivatives. LC-MS/MS can provide the analytical sensitivity, selectivity and precision required to enable accurate quantification of organic compounds in human biological liquid matrices. As a leading clinical LC-MS/MS solutions partner, Waters is a trusted provider of in vitro diagnostic medical devices, consumables, informatics, and support services for high performance LC-MS/MS systems. Join us at this workshop to gain insights into how our customers have addressed the challenging analysis of steroid hormones by implementing Waters solutions. LC-MS/MS Analysis of Steroids in the Clinical Laboratory Dr. Brian Keevil, University Hospital of South Manchester , Biochemistry Department Download Dr. Keevil's abstract http://www.waters.com/webassets/cms/events/media/EU%20Marketing/2018_MSACLEU_BK.pdf For Research Use Only. Not for use in diagnostic procedures. Learn more on Waters Solutions for LC/MS Analysis

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Corporate Workshops - Thursday 8:00 - 8:45

Zivak Technologies - Paracelsus Hall From Primary Sample Tubes to Results: World’s First Fully Automated Sample Preparation and Injection UHPLC System Zivak Technologies is delivering solution to manual sample preparation related problems of chromatography analysis and enabling clinical laboratories to make the most of their MS/MS instruments by full automatization. Fully automated Zivak analyzers allow you to get results from primary sample tubes without any user intervention. After the tubes are placed in the sample tray and the analysis is started on the computer, the robotic arms carry out all sample preparation steps, inject the prepared sample to MS/MS and chromatographic run starts. With Zivak analyzers, opening the sample tubes’ caps is never required. Attend the workshop and listen a detailed review of World’s First Walk Away Sample Preparation and Injection UHPLC System Multitasker and Zivak’s Revolutionary VD-200, Fully Automated Vitamin D2-D3 UHPLC Analyzer from Prof. Dr. Şahabettin Selek and Prof. Dr. Ahmet Ceyhan Gören.

Corporate Workshops - Thursday

13:30 - 14:15 Tecan - Papageno Hall Meet our experts in MS sample prep who will present innovative workflows using positive pressure workstations and liquid handling platforms to prep samples for clinical proteomics and steroid analysis. Semi-automated sample processing for clinical proteomics Stefan Loroch, Leibniz Institute for Analytical Sciences (ISAS), Proteomics Facility Powerful automated multi-steroid extraction via positive pressure and liquid handling platform for LC-MS analysis Dr. Christian Scherling, Tecan Germany, MS Sample Prep Application Specialist

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Podium Presentations

Wednesday & Thursday

Track Room Floor 1 Mozart 1-3 Ground 2 Mozart 4-5 Ground 3 Papageno Hall Ground 4 Paracelsus Hall 2nd 5 Trakl Hall 3rd 6 Doppler Hall 4th

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• Session 1 • Track 1 •

Lipidomics Wednesday @ 9:00 in Mozart 1-3

Session Chair: Julijana Ivanisevic - University of Lausanne Wednesday @ 9:00 in Mozart 1-3 Defining Boundaries of Lipidomic Variations in Human Cohorts Federico Torta - National University of Singapore -- *Young Investigator Grantee* ‣ Advances in mass spectrometry-based lipidomics have greatly expanded our understanding of the extent and complexity of lipid dysregulation in disease conditions. Targeted lipidomics enables the sensitive measurement of several hundred individual lipid species in thousands of samples for each study. We present here population-based studies aimed at clarifying the variation of plasma lipid concentrations in healthy individuals and in patients affected by metabolic disorders, in both cross-sectional and longitudinal studies. Wednesday @ 9:20 in Mozart 1-3 Shotgun Lipidomics of Cerebrospinal Fluid in Neurological Disorders Gábor Balogh - Biological Research Centre, Hungarian Academy of Sciences ‣ Biomarkers are urgently needed to improve diagnosis and prognosis of neurodegenerative diseases. Here we demonstrate that the lipid composition of cerebrospinal fluid (CSF) from Multiple Sclerosis, Alzheimer's disease (AD) and Guillain–Barré syndrome (GBS) patients shows significant signs of metabolic disturbances. In AD patients we found several lipid markers which were indicative to the severity of dementia, while the shift observed in the lipid profile of GBS patients supports the hypothesis of leakage from blood through the blood-nerve barrier. Therefore, lipidomic analysis of CSF might capable to reflect the pathophysiology of several neurological disorders. Wednesday @ 9:40 in Mozart 1-3 Understanding the Interplay Between Inflammatory Lipid Signaling and Demyelination Process: A Lipidomics Approach Applied to 3D Human Brain Model Maria Laura Santoru - University of Cagliari / University of Lausanne -- *Young Investigator Grantee* ‣ Demyelination and inflammation are two main features of multiple sclerosis. It has been proved that lipids play an important role in these two processes and to elucidate their mechanism of action in MS, a lipidomics approach was applied to iPSC-derived human 3D brain spheres system challenged by a treatment with cytokines to induce the demyelination. Combining targeted and untargeted lipidomics, we detected different classes of lipids in both spheres and their corresponding media. Among the measured lipid species, PAF and PAF related species were found to be significantly upregulated in the cytokine-treated spheres while the PUFA species and derived eicosanoids were increased in the spent media, suggesting a link between the early stage of inflammation process in MS and activated lipid signaling.

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Disease Implications and Diagnosis Wednesday @ 9:00 in Mozart 4-5

Session Chair: Grace van der Gugten - St Paul’s Hospital Wednesday @ 9:00 in Mozart 4-5 Simultaneous and Automated LC-MS Analysis of Plasma Methylmalonic Acid and Short Chain Fatty Acids Using a Carbodiimide-based Derivatization Mark Pratt - University Medical Center Groningen -- *Young Investigator Grantee* ‣ Methylmalonic acid has long been regarded as the most sensitive and specific functional marker of vitamin B12 status. However, it may not tell the complete story. Bacteria in the small intestine use vitamin B12 as a cofactor for the metabolism of succinic acid to propionic acid. Therefore, if vitamin B12 is utilized by bacterial overgrowth in the ileum, a vitamin B12 deficiency may occur despite sufficient dietary intake. Furthermore, this may result in high concentrations of the cytotoxic propionic acid. We have developed a novel LC-MS method for the simultaneous quantification of methylmalonic acid, propionic acid, succinic acid and butyric acid using a carbodiimide-based derivatization. This method may provide increased insight into the cause of vitamin B12 deficiencies, allowing for improved treatment strategies. Wednesday @ 9:20 in Mozart 4-5 Quantitation of Trimethylamine N-Oxide and its Precursors by HILIC-MS/MS and Their Potential Role in Cognitive Behavior Alexander Gaudl - Leipzig University ‣ Trimethylamine N-oxide (TMAO) enhances thrombocyte activity, affects cholesterol and bile acid metabolism and has already been associated with cardiovascular disease. Its role regarding cognitive behavior, however, remains open. To that matter and to establish normal concentration ranges we developed a new HILIC-MS/MS method for the quantitation of TMAO and its precursors Betaine, Carnitine, and Choline and applied it to a subset of the LIFE Adult study. A potential association with cognitive behavior was assessed using SISCO score. Wednesday @ 9:40 in Mozart 4-5 The Singapore Lipidomics Incubator: A Model for Engagement and Translation Anne Bendt - NATIONAL UNIVERSITY OF SINGAPORE ‣ We developed a model of engagement to clinically translate LC-MS/MS-based analytical protocols to measure metabolites (lipids, small molecules) in human plasma. By close interactions with data scientists, clinicians and laboratory medicine, we are translating some of our lab-based technology into assays suitable for clinical applications. A special focus lies in understanding natural variation and population reference values. Data from ongoing studies in multi-ethnic cohorts will be discussed.

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• Session 1 • Track 3 • Proteomics Keynote

Wednesday @ 9:00 in Papageno Hall Session Chair: Christa Cobbaert - Leiden University Medical Center

Wednesday @ 9:00 in Papageno Hall KEYNOTE PRESENTATION Clinical Proteomics: The Path Towards Implementation Harald Mischak - mosaiques therapeutics GmbH & University of Glasgow ‣ Clinical proteomics, the application of proteome analysis to clinical purpose, represents a major field in the area of proteome research. The aim of clinical proteomics is the improvement of clinical care based on (1) the identification and application of biomarkers, and (2) the suggestion of relevant therapeutic targets. These two areas have different requirements regarding specimens to be employed, technology, and data evaluation. While substantial efforts (especially in biomarker discovery, but also in the identification of therapeutic targets) are evident based on the large number of associated publications, only a few approaches have actually resulted in clinical application. In this presentation, key issues and major challenges in clinical proteomics will be discussed.

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New Technologies Wednesday @ 9:00 in Paracelsus Hall

Session Chair: Vladimir Frankevich - National Research Center for Obstetrics and Gynecology Wednesday @ 9:00 in Paracelsus Hall Probing Amyloid Aggregation Dynamics in vivo, Using SILK Imaging (iSILK) Jörg Hanrieder - University of Gothenburg -- *Young Investigator Grantee* ‣ Although the importance of amyloid beta (Aβ) plaque deposition in Alzheimer’s disease (AD) has long been recognized, exactly how plaques develop over time and their neurotoxic potential is not clear. Recent advances in imaging technologies such as imaging mass spectrometry (IMS) greatly increase the resolution of such events and the advent of isotope labelling of proteins and high-resolution IMS opens up possibilities for measuring spatial protein turnover kinetics in tissue. The aim of this study is to take advantages of such advances by using IMS along with metabolic labeling to recently developed novel genetic mouse models of AD. The results show distinct rates of secretion and deposition for distinct Aβ peptides within different brain regions. These data allow us to understand the progression of plaque deposition from initial seeding through to later aggregation. Wednesday @ 9:20 in Paracelsus Hall Identifying Biomarkers in FFPE Breast TMAs Using DESI-MSI Ólöf Gerdur Ísberg - University of Iceland -- *Young Investigator Grantee* ‣ Breast cancer is one of the most common cancers in the world among females and accounts for 25% of all cancers. DESI-MSI is a powerful tool to investigate the spatial distribution of biomolecules in tissue sections. The distribution of biomolecules such as metabolites can be correlated with clinicopathological information of tissue samples and thus providing essential information for clinical diagnosis. The project aim was to metabolic phenotype FFPE TMA slides of 30 breast cancer samples and 30 normal samples using DESI-MSI. Preliminary results report that DESI-MSI can discriminate between malignant and normal tissue. Wednesday @ 9:40 in Paracelsus Hall MALDI Imaging Through Laser-Induced Post-Ionization Pushes the Boundaries of Pharmaceutical Development Studies Florian Barré - M4I, Maastricht University -- *Young Investigator Grantee* ‣ Drug imaging is one of the major mass spectrometry imaging (MSI) applications. However, drug development through MSI has some limitations. Indeed, some compounds are not easily detectable due to poor ionization efficiency and ion suppression effects. In this work, we used post-ionization matrix assisted laser desorption/ionization (MALDI-2) to overcome these problems and we are showing that 80% of the drugs we tested directly benefit from ionization enhancement. We are now able to detect pharmaceuticals that are non-detectable by conventional MALDI pushing the boundaries of MSI for drug studies.

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Endocrinology Wednesday @ 9:00 in Trakl Hall

Session Chair: Flaminia Fanelli - University of Bologna Wednesday @ 9:00 in Trakl Hall Evaluation of Age and Metabolic Status Impact on Serum Steroid Hormones and Product/Precursor Ratios for the Generation of LC-MS/MS Reference Interval in Men Marco Mezzullo - University of Bologna -- *Young Investigator Grantee* ‣ Aiming at generating the reference intervals (RIs) for steroid levels and product/precursor ratios, we selected 321 adult drug- and disease-free Italian males, and defined a sub-cohort of 137 metabolically healthy individuals. Twelve serum steroids were measured by two validated LC-MS/MS assays. The independent impact of age, adiposity and metabolic parameters on steroid values was estimated, then, RIs at age 20, 30, 40, 50, 60, 70 and 80 were calculated in the overall and in the metabolically healthy cohort. We provided age- and metabolic status-specific RIs for a broad steroid profile, useful to implement LC-MS/MS in the management of endocrine diseases. Wednesday @ 9:20 in Trakl Hall Quantification of Testosterone, Androstenedione and 17-Hydroxyprogesterone Collected Using Mitra Micro Sampling Devices David Marshall - Manchester University NHS Foundation Trust -- *Young Investigator Grantee* ‣ Mitra devices are small polymer tips that absorb a fixed amount of blood which can be dried prior to analysis. A simple liquid:liquid extraction was used following reconstitution of the mitra device in distilled water. Separation was carried out using a Waters Acquity HSS T3 column which resolved the peaks well. We have developed an assay for the simultaneous quantification of testosterone, androstenedione and 17-hydroxyprogesterone using a single mitra tip. Wednesday @ 9:40 in Trakl Hall Direct Analysis - No Sample Preparation - of Bioavailable Cortisol in Human Plasma by Weak Affinity Chromatography (WAC; Affinity LC/MS) Sten Ohlson - Nanyang Technological University ‣ Pre-analytical treatment of blood plasma is a time consuming and often rate limiting step in the workflow of LC/MS analysis. We present in this study a new approach for quantitative LC/MS based clinical analysis based on Weak Affinity Chromatography (WAC) making sample preparation obsolete.

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Practical Training : Proteomic Assay Development Wednesday @ 9:00 in DopplerHall

Session Chair: Tim Collier - Quest Diagnostics - Cleveland Heart Lab Wednesday @ 9:00 in Doppler Hall Proteomic Assay Development: Digest Optimization and Quality Control Timothy Collier - Quest Diagnostics - Cleveland HeartLab ‣ The development of protein mass spectrometric assays for use in the clinical laboratory presents unique workflow challenges. Specific pre-analytical steps, such as enzymatic proteolytic digestion, are often required to generate peptides amenable to mass spectrometric analysis. This process requires close attention during and after assay development to ensure reproducible and accurate clinical analysis. This practical training session will focus on Digest Optimization, including enzyme selection, reagent systems to boost efficiency, and quality control mechanisms to allow monitoring of digest performance. Additional quality control mechanisms to monitor assay performance will also be discussed, including secondary peptide and ion-ratio monitoring, enrichment process monitoring, and steps to ensure reagent quality.

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Metabolomics Methodology Wednesday @ 11:00 in Mozart 1-3

Session Chair: Elizabeth Want - Imperial College London Wednesday @ 11:00 in Mozart 1-3 New Insights in CE-MS-based Metabolomics Nicolas Drouin - University of Geneva ‣ During this study, we developed an original workflow for features identification in CE-MS. This innovative approach is based on the conversion of migration time scale into an effective mobility (µeff) scale of data acquired in standardized conditions and an experimental µeff based library of more than 400 endogenous compounds. This method allowed lab-to-lab identification of features from cell cultures. Wednesday @ 11:20 in Mozart 1-3 Combined Targeted and Untargeted Strategy for Quantification of Amino Acids and Acylcarnitines in Clinically Relevant Biofluids and Tissue Lysates Vera van der Velpen - Metabolomics Unit, University of Lausanne -- *Young Investigator Grantee* ‣ Amino acids and acylcarnitines are key players in energy metabolism, however, analytical methods for comprehensive and robust measurement of these compounds without derivatization or use of ion-pairing agents are scarce. We present a HILIC-based HRMS method for the absolute quantification of amino acids and acylcarnitines in a single run, while simultaneously taking advantage of HRMS in full scan mode to screen for additional derivatives (e.g. amines, hydroxylated acylcarnitines) and other polar metabolites (e.g. purines, pyrimidines, hexoses). Using a simple extraction method with internal standards, we have successfully evaluated our method with NIST plasma and examined a profile of these compounds across different biofluids (plasma, urine, CSF) and human brain tissue. Wednesday @ 11:40 in Mozart 1-3 Development of a Pipeline for Automated Reconstruction and Annotation of Data Independent Acquisition LC-MS Datasets Gonçalo Graça - Imperial College London -- *Young Investigator Grantee* ‣ Reconstruction of parent-fragment ion relationships from data independent acquisition (DIA) remains a challenge in non-targeted MS metabolomics and lipidomics analysis. In the presented work we propose a novel strategy to improve the reconstruction of DIA fragmentation MSE data in order to automatically annotate the majority of known metabolites/lipids in human serum.

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All About Breath Analysis Wednesday @ 11:00 in Mozart 4-5

Session Chair: Neus Fabregat-Cabello - University of Liege Wednesday @ 11:00 in Mozart 4-5 Mass Spectrometry Applied to Large-Scale Multi-Centre Clinical Project in Breath Research Ilaria Belluomo - Imperial College London -- *Young Investigator Grantee* ‣ Non-invasive and rapid diagnostic testing, such as breath testing, is the future of diagnostic medicine. We are establishing a workflow for a multi-centre large-scale clinical project for the diagnosis of colorectal cancer. This involves analysis of breath samples with two different mass spectrometry platforms, requiring reliability and a strong quality control system. The use of two different but complementary technologies and the three levels of quality control for this large-scale screening increased sensitivity, specificity and reliability of the results. Thanks to our multi-instrument platform and three different levels of quality control, we have an accurate, specific and sensitive method for determining volatile organic compounds profile in human breath. Wednesday @ 11:20 in Mozart 4-5 Cross Platform Validation of Mass Spectrometry for the Diagnosis of Colorectal Cancer Using a Breath Test Georgia Woodfield - Imperial College London -- *Young Investigator Grantee* ‣ A breath test for colorectal cancer (CRC) could be a valuable non-invasive diagnostic tool. The Colorectal BReath Analysis (COBRA) prospective study aims to determine the accuracy of breath volatile organic compounds (VOCs) for detecting CRC, building on previous literature. This crucially depends on accurate breath profiling. We performed cross platform validation of breath VOCs from the first 406 patients using gas chromatography mass spectrometry vs. proton transfer reaction mass spectrometry. This proved to be an accurate, reliable method for identifying and quantifying VOCs, with good inter-instrument agreement. This widespread use of a breath test for CRC seems technically possible. Wednesday @ 11:40 in Mozart 4-5 A Pilot Study Profiling Volatile Organic Compounds (VOCs) in the Breath of Patients with CRC Using Proton Transfer Mass Spectrometry Harrypal Panesar - Imperial College London ‣ We initially asked the question: Is it possible to distinguish between health patients and those with colorectal cancer (CRC) using their breath profiles? Previous research has suggested differences in VOC profile between health and CRC patients using Selected Ion Flow Tube Mass Spectrometry (SIFT-MS). However, we employed a new technique, Proton Transfer Mass Spectrometry (PTR-MS), to measure the VOC profile in the breath of CRC and healthy patients. Analysing the breath profile of 17 patients with CRC and comparing them to normal colons, we describe a profile of breath VOCs that are significantly different between normal and CRC patients.

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Endocrine Keynote Wednesday @ 11:00 in Papageno Hall

Session Chair: Brian Keevil - Manchester University Wednesday @ 11:00 in Papageno Hall KEYNOTE PRESENTATION Mass Spectrometry Harmonisation: Paediatric Steroid Tales Ronda Greaves - Murdoch Children's Research Institute ‣ Our goal in laboratory medicine is to improve patient health by using laboratory tests as our tool to support medical decisions. The change from immunoassay to mass spectrometry analysis of steroids has significantly improved the specificity, accuracy and sensitivity of many steroids frequently analysed in children. However, mass spectrometry assays are not infallible and recognised differences exist across the total testing process. There are demonstrated variations pre-analytically, analytically and post-analytically in methods and understanding what differences are critical and how results compare between laboratories is essential to close the gap between result differences between laboratories. In this presentation, we will explore the challenges and possible solutions for the harmonisation of paediatric steroid analysis by mass spectrometry to support medical decisions.

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Glycomics 1 Wednesday @ 11:00 in Paracelsus Hall

Session Chair: Guinevere Lageveen-Kammeijer - Leiden University Medical Center Wednesday @ 11:00 in Paracelsus Hall Global Glycoproteomic Analysis of O-glycosylation in Human Herpesviruses Ieva Bagdonaite - University of Copenhagen ‣ Information on site-specific O-glycosylation of viral envelope glycoproteins is generally very limited despite important functions. We present a powerful mass-spectrometry based strategy to globally identify O-glycosylation sites on viral envelope proteins of a given virus in the context of a productive infection. We successfully utilized the strategy to map O-linked glycosylation sites on several complex herpesviruses demonstrating that O-glycosylation is widely distributed on most envelope proteins. Moreover, we used genetically engineered keratinocytes lacking O-glycan elongation capacity to demonstrate that O-linked glycans are indeed important for HSV-1 biology as HSV-1 particles produced in these cells had significantly lower titers compared to wild-type keratinocytes. These tools enable wider discovery and detailed analysis of the role of site-specific O-glycosylation in virology. Wednesday @ 11:20 in Paracelsus Hall In Depth O-Glycosylation Structural Analysis of Colorectal Cancer Cell Lines Katarina Madunic - Leiden University Medical Center -- *Young Investigator Grantee* ‣ Altered glycosylation has been widely described in cancer. However, little is known about O- glycosylation due to the complexity of the analytical approaches. Many studies used single cell lines to investigate glycosylation changes in cancer without taking into account the differences between cell lines. During this study O-glycosylation signatures of 25 colorectal cancer cell lines were examined by nano-PGC-LC-ESI-MS, allowing the differentiation between isomeric structures and their association with different cell phenotypes. Our results have shown profound differences between O-glycomes of the cell lines revealing that single cell line studies may not provide representative results. Wednesday @ 11:40 in Paracelsus Hall HT UPLC Profiling of Total Plasma N-Glycans in Type II Diabetes Mellitus from Patients and Healthy Individuals in a Ghanaian Population Elham Memarian - Leiden University medical centre -- *Young Investigator Grantee* ‣ Aberrant protein glycosylation may reflect changes in cell metabolism of type II diabetes mellitus (T2DM) and offers fresh vistas for discovering potential biomarkers. However, the functional significance of T2DM N-glycan alterations is underexplored, since to date N-glycan profiling studies have been mainly performed in selected populations. Geographically and genetically isolated populations are needed for validation of specific biomarkers.

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MS/MS for Proteotyping Pathogens Wednesday @ 11:00 in Trakl Hall

Session Chair: Simon Cameron - Imperial College London Wednesday @ 11:00 in Trakl Hall Phylopeptidomics: A Step Closer to Adopting the Microbiome Data into Clinical Practice Lucia Grenga - CEA -- *Young Investigator Grantee* ‣ By influencing a variety of host processes, the microbiome plays a crucial role in human health and disease. Here we present the application of an innovative tandem mass spectrometry -based approach to the study of the microbiome structure and dynamics. Named phylopeptidomics, this new strategy allows the rapid identification of the components of the microbiome, assessment of their biomass contributions, their deep functional characterization and the simultaneous identification of the resistance arsenal and/or toxin production of the present organisms. Coupled with metabolomic analyses, the phylopeptidomics methodology could drive, through a better understanding of the host-microbiome crosstalk, to the development of new diagnostic tools and/or therapeutic strategies. Wednesday @ 11:20 in Trakl Hall Characterization of the Virulome of Staphylococcus Aureus by a Highly Multiplex Approach with Scout-MRM Mouton Nicolas - University de Lyon Claude Bernard -- *Young Investigator Grantee* ‣ The emergence of antibiotic-resistant bacteria strains is seriously threatening human life, as recently underlined by the World Health Organization. The case of Staphylococcus aureus, S.a. is of particular concern, whose forms resisting to vancomycin or methicillin caused numerous patients death in Hospitals. It is also crucial to better understand how mutations of the bacterial genome induce the emergence of new resistant strains. The sensitivity and multiplexing possibilities of mass spectrometric (MS) approaches appears particularly relevant to tackle such problems. We thus developed a targeted assay covering 90 key proteins involved in S.a. virulence and antibiotic resistance using a novel and a highly multiplexed targeted-MS approach, Scout-MRM. The virulome assay has been implemented for deciphering the proteogenomic links across a clinical collection of 230 S.a. strains. Wednesday @ 11:40 in Trakl Hall Deciphering MALDI-TOF Identification of Bacteria Using a Proteogenomic Approach Frédéric Jauffrit - Université de Lyon -- *Young Investigator Grantee* ‣ Ribosomal and non-ribosomal proteins producing to peaks in WC-MALDI-TOF MS spectra were identified using proteogenomics and, as a case study, the VITEK® MS calibrant strain, Escherichia coli ATCC8739. Protein expression was evidenced using LC-MS/MS data and a 6 frame translation of the entire genome. ORFs were inferred and MALDI-TOF peaks were assigned to possible protein m/z according to gene sequences, PTMs and charge states. This method, applied on 832 E. coli spectra, allowed the identification of all major MALDI-TOF peaks (56 peaks above 3% relative intensity) and more deeply of 424 proteins corresponding to 649 peaks.

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Practical Training : Calibrators and Internal Standards for Protein MS Wednesday @ 11:00 in Doppler Hall

Session Chair: Chris Shuford - Lab Corp Wednesday @ 11:00 in Doppler Hall Calibrators and Internal Standards in Protein Mass Spectrometry Assays: Differences, Commonalities, and Best-Practice Christopher Shuford - Laboratory Corporation of America ‣ Internal standards are not calibrators. This fact is often overlooked in the field of proteomics, which has propagated misconceptions as new protein mass spectrometry assays are translating into the clinical chemistry lab. In the first part of this 3 part series, the distinction between calibrators and internal standard will be highlighted in the context of protein measurements by mass spectrometry. In the second part of the series, best practices for calibration and standardization/harmonization of protein mass spectrometry assays will be discussed. Finally, options for internal standardization of protein mass spectrometry assays will be reviewed and contrasted in the third part of the series.

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Metabolic Markers Wednesday @ 14:30 in Mozart 1-3

Session Chair: Luke Whiley - Imperial College London Wednesday @ 14:30 in Mozart 1-3 Discovery of Novel Galabiosylceramide-Related Biomarkers of Fabry Disease by Semi-Targeted Metabolomics: the Complex Issue of Structural Isomer Interferences Michel Boutin - Université de Sherbrooke ‣ Fabry disease is an X-linked lysosomal storage disorder causing severe cardiac, renal and cerebrovascular complications. A metabolomic study targeting lipids in urine revealed 22 galabiosylceramide (Ga2) isoforms/analogs as Fabry disease biomarkers. Unfortunately, the efficiency of these biomarkers was significantly compromised by the co-analysis of their lactosylceramide (LacCer) structural isomers differing only by the conformation of one glycosidic linkage. A normal phase chromatography was developed to separate Ga2 isoforms/analogs from their LacCer counterparts. The removal of the LacCer interferences significantly increased the sensitivity of Ga2 biomarkers, especially for untreated Fabry females (from 9.3% to 70.4%.). Wednesday @ 14:50 in Mozart 1-3 Identifying the Metabolic “Achilles Heel” of Adult and Childhood Brain Cancers Using LC-MS-based Metabolite Profiling Dong-Hyun Kim - University of Nottingham ‣ Glioblastoma multiforme (GBM) was recently shown to be dependent on extracellular sources of lipids to meet cellular requirements as opposed to de novo biosynthesis. We conducted LC-MS-based metabolomics to characterise the cell metabolome of GBM cell lines cultured under lipoprotein-deficient conditions to replicate nutrient stresses seen in vivo. As a result, cell line-specific metabolite changes were observed calling for the need for integration with transcriptomics data to elucidate genetic drivers of cell line-specific stress responses. This study has highlighted the metabolic dependencies of GBM under nutrient-limiting conditions which will inform on the development of new therapeutic strategies targeting stress responses. Wednesday @ 15:10 in Mozart 1-3 Urinary Biomarkers of Dietary Intake - Quantification of Sugars for Epidemiology Studies Using LC-MS/MS Nicola Gray - University of Reading -- *Young Investigator Grantee* ‣ For epidemiology studies that aim to investigate how diet influences health, accurate records of nutritional intake are essential. Nutritional assessment typically relies on self-reported food diaries or food-frequency questionnaires, known to be inaccurate and introduce bias. The development of accurate markers to assess dietary intake is therefore crucial to investigate associations between diet, health and disease. For example, urinary sucrose and fructose have been shown to correlate with total sugars intake and have been developed as a marker of sugar consumption. The application of such a biomarker to population studies require robust methods suitable for high-throughput analysis. Here, an LC-MS/MS method has been developed to distinguish and quantify sugars in urine for application to large clinical cohorts as an accurate measure of sugars consumption in free-living populations.

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Keynote Small Molecules Wednesday @ 14:30 in Mozart 4-5

Session Chair: Michael Vogeser - Medical University of Muenchen Wednesday @ 14:30 in Mozart 4-5 KEYNOTE PRESENTATION Tackling Analytical Challenges in Sports Drug Testing by Mass Spectrometric Approaches Mario Thevis - German Sport University, Cologne, Germany European Monitoring Center for Emerging Doping Agents (EuMoCEDA), Cologne/Bonn, Germany ‣ Sports drug testing laboratories are facing multifaceted challenges including the misuse of naturally/endogenously occurring substances, non-approved/discontinued drug candidates, urine manipulation, etc. In order to provide best-possible analytical performance, mass spectrometry-based approaches are predominantly utilized to detect prohibited substances and methods of doping. With the constantly increasing analytical requirements concerning the number of target compounds, the complexity and range of physico-chemical properties of analytes (e.g., inorganic ionic transition metals, gases, lipids, alkaloids, peptides, proteins, DNA/RNA-based drugs, etc.) as well as the desire to accelerate analyses and obtain information allowing also for retrospective data mining, high resolution/high accuracy mass spectrometry has become a mainstay in doping controls. In that context, various assays have been reported, enabling either multi-component analyses of low- or high molecular mass measurands or the specific and dedicated (confirmatory) detection of prohibited substances. Selected applications will be presented reporting on examples of recent findings in routine sports drug testing, demonstrating both the inventiveness of cheating individuals that undermine current anti-doping efforts as well as the relevance of in-depth investigations into unusual findings, where the athletes’ innocence was to be shown albeit prohibited substances were unequivocally identified in their doping control urine samples.

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Cardiovascular and Thrombotic Disease Wednesday @ 14:30 in Papageno Hall

Session Chair: Tim Collier - Quest Diagnostics - Cleveland Heart Lab Wednesday @ 14:30 in Papageno Hall Building Standards for Proteomics: A Targeted Mass Spectrometry Approach for Quantification of Cardiovascular Disease Biomarker in Human Blood Albert Sickmann - ISAS ‣ Platelets are primarily known to be key players in thrombosis and hemostasis. Targeting platelet function and signaling may represent novel therapeutic strategies in prevention of cardiovascular diseases. The absolute quantification of phosphorylation sites in actived platelets can direct the establishment of new diagnostic assays by characterization of samples with clinical relevance, including quantification of platelet receptors and signaling proteins and their regulation via posttranslational modifications in human blood. Wednesday @ 14:50 in Papageno Hall Validation of ApolipoProtein A-I Associated Lipoprotein Panel for the Prediction of Cholesterol Efflux Capacity Cory Bystrom - Cleveland HeartLab ‣ We developed a rapid affinity method to enrich apolipoprotein A-I associated lipoproteins and utilized mass spectrometry-based approach for multiplex quantitation. Utilizing this technique, a multiprotein panel was developed which can be used to estimate cholesterol efflux capacity. The analytical workflow was validated and used in an 8 week longitudinal study. In normal healthy volunteers, predicted cholesterol efflux was stable. Wednesday @ 15:10 in Papageno Hall Defining the Measurands for Antithrombin and its Proteoforms Using Mass Spectrometry Based Techniques Renee Ruhaak - Leiden University Medical Center ‣ Current medical tests for antithrombin deficiency generally measure the overall activity, being blind for the actual proteoforms of AT that contribute to the test. Here, we present an MRM-based test that allows for the identification and quantification of clinically relevant proteoforms of AT according to predefined analytical performance specifications, as well as the characterization of AT candidate reference materials using high-end proteomics strategies. This now enables further unravelling of the individual contribution of the different AT-proteoforms to AT activity test results, which is a first and crucial step in understanding the value of the current AT tests offered in the clinical laboratory.

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Tissue Imaging Keynote Wednesday @ 14:30 in Paracelsus Hall

Session Chair: Tiffany Porta - Maastricht University Wednesday @ 14:30 in Paracelsus Hall KEYNOTE PRESENTATION Combining Mass Spectrometry Imaging & Microproteomics to Investigate Intratumor Heterogeneity Liam McDonnell - Fondazione Pisana per la Scienza ONLUS, Pisa, Italy ‣ Mass spectrometry imaging (MSI) is able to simultaneously record the distributions of hundreds of molecules directly from tissue. This spatially-resolved molecular information can be combined with multivariate/clustering analysis to reveal regions of tissue with distinct molecular signatures, a process that has been termed MSI-based molecular histology and has been used to reveal tumor subpopulations, metabolically distinct cell layers, and tumor interface zones. Rapid direct tissue analysis is essential for MSI in order to maintain spatial localization and acceptable measurement times. The absence of an explicit analyte separation/purification step means MSI lacks the depth of coverage of LC-MS/MS. Here, we demonstrate how MSI can be combined with high sensitivity microproteomics, even of the same tissue section, to further investigate the molecular changes associated with tumor subpopulations.

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Glycomics 2 Wednesday @ 14:30 in Trakl Hall

Session Chair: TBD Wednesday @ 14:30 in Trakl Hall In-Depth Characterisation of Heavily Glycosylated Proteins and Their Potential as Colorectal Cancer Biomarkers Valeriia Kuzyk - Vrije Universiteit Amsterdam -- *Young Investigator Grantee* ‣ Protein glycosylation is known to rapidly change in disease, as well as in cancer progression. Therefore, heavily glycosylated tumor-derived circulating proteins hold a great potential to perform as “disease ambassadors” in the bloodstream, enabling more sensitive and specific diagnostics. However, proper characterization is challenging due to low quantity in serum and the high heterogeneity of the glycan distribution. Here, we present a comparison between different analytical platforms for the in-depth characterization of two glycoproteins (carcinoembryonic antigen and laminin B1). Further studies will examine if altered glycosylation profiles of these glycoproteins can be used as prognostic tools for colorectal cancer. Wednesday @ 14:50 in Trakl Hall Characterisation of Prostate-Specific Antigen Isolated from Patients’ Urine Alan Moran - Leiden University Medical Centre -- *Young Investigator Grantee* ‣ The glycoprotein prostate specific antigen (PSA) is a clinical biomarker of prostate cancer (PCa) but has a rather poor specificity and predictive value limiting its value for PCa diagnosis. It has been hypothesized that through examining its glycosylation, PSA could be used as a more specific biomarker of PCa. This study presents a novel approach to analyse the glycosylation variants as well as other proteoforms of PSA from patients’ urine using capillary electrophoresis coupled to a mass spectrometer with a sheathless interface (CE-ESI-MS). Analysis of intact PSA revealed a wide variety of glycoforms and proteoforms and results obtained from intact and glycopeptide analysis were compared. Wednesday @ 15:10 in Trakl Hall Development of MS-based Prostate-Specific Antigen Test with In-Depth Glycosylation Analysis Guinevere S.M. Lageveen-Kammeijer - Center for Proteomics and Metabolomics, LUMC -- *Young Investigator Grantee* ‣ The diagnostic test for prostate cancer (PCa) based on glycoprotein prostate-specific antigen (PSA) is widely known for its lack in specificity. Recently, we established a PSA glycomics assay that allows identifying and relatively quantifying the glycosylation profile of PSA in a non-invasive manner after affinity capturing from patients’ urine. The applicability of this assay is further explored for other biofluids such as seminal plasma and serum. Potentially, alterations in the glycosylation pattern will provide better diagnostic and prognostic tools for fertility conditions (seminal plasma) and PCa (serum and urine) by studying different cohort studies.

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Practical Training : EU Regulations Wednesday @ 14:30 in Doppler Hall

Session Chair: Brian Keevil - University of Manchester Wednesday @ 14:30 in Doppler Hall EU LC-MS Verification Brian Keevil - Manchester University ‣ The new EU regulations for the use of diagnostic kits will soon come into force. How will this affect laboratories and how will the laboratories verify the results produced by these kits? The UKAS standard used in UK laboratories (ISO 15189) is not prescriptive, it tells us what we should be doing, but not how to do it. To help us in this there are many guidelines for the validation /verification of kits. However, some of these guidelines are more prescriptive than others and give greater detail on what needs to be done. The talks will focus on the techniques used locally to validate and monitor LC-MS assays and verify currently available diagnostic kits.

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Metabolomics – Clinical Studies I Thursday @ 9:00 in Mozart 1-3

Session Chair: Oleg Mayboroda - Leiden University Medical Center Thursday @ 9:00 in Mozart 1-3 Metabolomic Analysis of Changes in the Serum of Diabetic Patients Caused by Ulcers Arsenty Melnikov - Internatrional Tomography Center SB RAS -- *Young Investigator Grantee* ‣ Patients with diabetes mellitus have approximately a 20% risk of developing an ulcer, 7% cases of which leads to an amputation. Despite widespread occurrence of diabetic ulcers, this complication is poorly studied, making this disease a topical object of study. Current report provides the data on metabolomics changes in the serum of diabetic patients without complications as compared to the serum of diabetic patients with ulcers. More than 200 metabolites were detected with the use of two independent HPLC-MS method (HILIC and RPLC). The concentrations of several metabolites are found to differ significantly: phospholipids, bilirubin, 1,5–anhydroglucitol, oleamide. Thursday @ 9:20 in Mozart 1-3 A Metabolite Level View of the Gut-Brain Axis in Dementia and Aging: Quantification of the Kynurenine/Tryptophan Pathway Using Mass Spectrometry Luke Whiley - Imperial College London -- *Young Investigator Grantee* ‣ Compositional variation in the gut microbiota has been reported in dementia and aging. The intestinal bacteria are now known to influence biochemical processes in the brain and modify host behaviours and cognitive development. This gut-brain axis arises through a variety of mechanisms that are not completely understood. One mechanism is the tryptophan-kynurenine pathway. Intestinal bacteria can metabolize tryptophan, a precursor for serotonin, reducing its bioavailability. In addition, microbes can interact with the immune system altering the amount tryptophan converted to kynurenine and its neuroactive metabolites. A UPLC-MS method was developed to quantify serum/plasma metabolites in this pathway and assess their variation in large clinical cohorts reflective of neurological disease, cognition and dementia. Thursday @ 9:40 in Mozart 1-3 Estimation of the Post Mortem Interval via Metabolomic Approach Lyudmila Yanshole - International Tomography Center SB RAS -- *Young Investigator Grantee* ‣ The analysis of post-mortem metabolomic changes in biological fluids opens the way to develop new methods for the estimation of post-mortem interval (PMI) which appears to be an actual problem in forensic medicine. The combination of modern research methods - high frequency NMR and high-performance liquid chromatography with high-resolution mass spectrometry (HPLC-MS) was used in this work. Metabolomic profiling of rabbit and human biological fluids obtained at different times after death is performed in this work: the quantitative levels of 61 metabolites have been measured. The obtained results suggest that the ocular fluids AH and VH may have some advantages over blood serum for the search of potential biochemical markers for the PMI estimation. Among the compounds studied in the present work, hypoxanthine, choline and glycerol give the biggest promise as the potential PMI biomarkers.

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Quality Issues Thursday @ 9:00 in Mozart 4-5

Session Chair: Anne Bendt - National University of Singapore Thursday @ 9:00 in Mozart 4-5 Quality Assurance and Quality Control in LC-MS-based Targeted Lipidomics of Human Blood Plasma Bo Burla - National University of Singapore -- *Young Investigator Grantee* ‣ Ensuring quality and reproducibility of large-scale, large-panel LC-MS-based lipidomics analyses is critical but still challenging. Experimental design, sample processing, matrix effects, instrumental variations and data processing can all introduce variations and inter- and intra-assay batch effects, which can bias or cofound experimental readouts. We here explore frequently used and novel quality assurance (QA) and quality control (QC) strategies based on internal standards, different QC sample types, reference materials and different corresponding data processing methods and software tools. We demonstrate these strategies with analyses of large-scale studies and specifically designed tests. Thursday @ 9:20 in Mozart 4-5 Stable Isotope-Dilution-LC-MS/MS-based Reference Methods – Suggestion for Description of Methods and Structure of Analytical Series Michael Vogeser - University Hospital, LMU Munich ‣ ID-LC-MS/MS methods are increasingly used as reference for routine methods, but actually the characteristics of such reference methods (RM) are so far not conclusively specified. A RM concept specifically for ID-LC/MS was developed, including three key elements: 1. Standardized method description – differentiating between essential characteristics (such as recorded mass transitions) and variable features that realistically can not be standardized over time (such as lot of columns). 2. Standardized structure of analytical series, including several features for performance verification (e.g., addressing matrix effects). 3. Based on this, a series-related validation concept according to pre-defined criteria as essential characteristic of the RM. This novel approach was successfully applied to a RM for the quantification of linezolid in serum. Thursday @ 9:40 in Mozart 4-5 Development of an Assay for Citrate and Oxalate in Urine by Hydrophilic Interaction LC-MS/MS After Weak Anion Exchange SPE Albert Dijkhuizen - Leiden University Medical Center ‣ Traditionally urinary oxalate and citrate were analyzed with enzymatic and colorimetric methods. Today chromatographic-mass spectrometric methods become more common. We developed a HILIC LC-MS/MS method including SPE for the determination of oxalate and citrate. After sample clean-up with a Waters Oasis WAX SPE-column, extracts are injected on a Waters BEH-amide column. Elution is performed with a linear acetonitrile to water (pH=10.0) gradient. Retention-times are 2.1 and 2.8 min. for oxalate and citrate respectively. Calibration-curves are linear from 10-1000 µM for oxalate and 50-5000 µM for citrate. The method is free of interfering peaks and ion suppression.

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Clinical Chemistry I Thursday @ 9:00 in Papageno Hall

Session Chair: Éva Hunyadi-Gulyás - Biological Research Center Thursday @ 9:00 in Papageno Hall A Newly Developed LC-MS/MS Method for the Quantitation of Glucagon in Plasma Shows the Lack of Specificity of Former RIA Methods Jordi Farre-Segura - University of Liège, CHU de Liège -- *Young Investigator Grantee* ‣ We have developed a sensitive method for intact glucagon quantitation by LC-MS/MS based on a simple solid phase extraction. The method showed good precision, accuracy and detection limits throughout its advancement. However, our preliminary comparison with our routine radioimmunoassay (RIA) showed a significant bias. Hence, RIA provides spurious results compared to the LC-MS/MS that could suppose a change in the perspective of this peptide’s measurement. Thursday @ 9:20 in Papageno Hall Progress in Clinical Applications for the MALDI-Imaging Analysis of Thyroid Biopsies: Evaluation of Proteomic Stability in Preservative Solutions Isabella Piga - University of Milano-Bicocca -- *Young Investigator Grantee* ‣ Thyroid lesions diagnosis is performed by image-guided fine needle aspiration biopsy (FNAB) regarded as a gold standard procedure. However, about 25% of FNABs are considered as “indeterminate for malignancy” and surgery is commonly recommended. Nevertheless, final histology highlight that 80% of them are benign lesions. Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) enables to explore the spatial distribution of biomarkers directly on cytological specimens, by integrating molecular and morphological evidence. This study focus on the proteomic profile stability of cytological samples in well-known preservative solutions, in order to have a standard methodology for the proteomic MALDI-MSI analysis of thyroid FNABs. Thursday @ 9:40 in Papageno Hall Selective Determination of Human and Synthetic Insulins in Plasma Using LC-MS – How Does it Help the Clinician? Jean-Christophe Prost - Bern University Hospital (Inselspital) ‣ Quantification of insulin in blood is of high importance for diagnostic, therapeutic, forensic and research purposes. Although commercially available immunoassays are very sensitive, but they lack selectivity between insulin and insulin analogues. The recent introduction of immunoaffinity workup coupled to ultra high-performance liquid chromatography mass spectrometry offers a promising approach for the simultaneous quantification of endogenous and synthetic insulins. The assay developed on our triple quadrupole mass spectrometer demonstrated good sensitivity and selectivity for endogen and exogen insulins, and could help clinician along avoiding several immunoassay analysis to monitor selectively multiple insulins by insulin-treated diabetes patients.

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MSI for Clinical Applications Thursday @ 9:00 in Paracelsus Hall

Session Chair: Raf Van De Plas - Delft Univerity of Technology Thursday @ 9:00 in Paracelsus Hall Next Generation Sequencing After MALDI MSI – A Feasibility Study Kristina Schwamborn - Institute of Pathology, TU Munich ‣ Limited amount of sample material is an inherent problem in early cancer diagnostics. Thus, developing strategies to enable multiple test to be performed on the same tissue section are needed. Combining MALDI MSI and next generation sequencing using only a single tissue section is feasible and could be of value in cases of limited sample material. Thursday @ 9:20 in Paracelsus Hall MALDI-MSI in the Search for Proteomic Indicators of Response to Therapy in Membranous Nephropathy Andrew Smith - University of Milano-Bicocca -- *Young Investigator Grantee* ‣ Here we demonstrate how high spatial resolution MALDI-MSI can assist in the search for proteins able to predict response to treatment in Membranous Nephropathy patients. We present three proteins, macrophage migration inhibitory factor (MIF), Sonic Hedgehog (SHH) and α-smooth muscle actin (α-SMA), whose signal intensity and spatial localisation differed between patients who responded favourably or unvaourably to therapeutic treatment (Ponticelli Regimen). If verified in a larger sample cohort, such findings may have direct clinical relevance and could provide support in the prognostic assessment of MN patients. Thursday @ 9:40 in Paracelsus Hall Bringing Mass Spectrometry to Pathologists for Diagnostics of Frozen Surgically Resected Tissue Specimen Tiffany Porta Siegel - M4I, Maastricht University ‣ In current clinical practice, accurate tumor diagnosis during surgery remains challenging. To assess the presence of a malignancy, frozen cut tissue sections are evaluated by the pathologist; but this remains challenging because the morphology of the tissue altered. With our approach, we aim for a rapid, on-line solution by combining an optical microscope directly to a mass spectrometer that could help the pathologist with molecular evaluation / diagnostics of fresh tissue based on molecular profiling within minutes. A predictive statistical model based on PCA-LDA analysis of the patient specimens displayed an accuracy of >90% correct classification rate which allowed to differentiate tumor versus non tumor tissues from patients undergoing surgery for colorectal metastasis. This holds great promises for the diagnostics of frozen section for intraoperative tumor diagnostics.

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Keynote for Microbiology Thursday @ 9:00 in Trakl Hall

Session Chair: Jean Armengaud - CEA-Marcoule Thursday @ 9:00 in Trakl Hall KEYNOTE PRESENTATION Tandem MS-Proteotyping: Proteomics- and Genomics-based Characterization and Typing of Infectious Bacteria Edward Moore - University of Gothenburg ‣ LC-MS/MS proteomics- and genomics-based characterization and typing of microorganisms, i.e., ‘proteotyping’, using peptides digested from expressed proteins and matched to genomic sequence data, can be applied for sensitive and accurate detection and typing of pathogenic bacteria. Proteotyping is capable of resolving and identifying closely-related bacterial taxa with simultaneous detection of virulence and antibiotic resistance features, providing for comprehensive characterizations of infectious bacteria. The methodology may be applied directly to analyses of clinical samples without prior cultivation and isolation, thus providing for rapid, reliable, infectious disease diagnostics.

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Practical Training : Finding Your Analyte Thursday @ 9:00 in Doppler Hall

Session Chair: Catarina Horro-Pita - LGC Thursday @ 9:00 in Doppler Hall Where Did My Analyte Go? – coping with poor solubility and non-specific binding Catarina Horro Pita - LGC, Drug Development Solutions ‣ Poor solubility and non-specific binding can be insidious problems that are often overlooked during method development. These issues can result in costly and time-consuming batch failures during validation and, more importantly, the generation of inaccurate results during sample analysis. This training will describe in detail these issues, illustrate case studies and conclude with a workshop, where the audience will be asked to review ‘troublesome’ methods and suggest possible solutions.

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Metabolomics Clinical Studies II Thursday @ 11:00 in Mozart 1-3

Session Chair: Nicola Gray - University of Reading Thursday @ 11:00 in Mozart 1-3 Analytical Strategies to Study the Interaction Between Microbiota and Drug Metabolism Using Targeted and Untargeted LC-MS-based Metabolomics Marine Letertre - Imperial College London -- *Young Investigator Grantee* ‣ Metabonomics, and particularly pharmacometabonomics, is a useful tool for patient stratification and to establish individualized drug therapy and limited ADRs to improve patient journey. We have used the pharmacometabonomics principle on animal experiments to study the interaction between the microbiome and several drugs. The general workflow applied both untargeted LC-MS for endogenous metabolic phenotyping and a targeted approach to quantify the drug of interest and its metabolites, in complement of NMR and amplicon sequencing. It enabled the impact of drugs on the gut microbiota community structure to be investigated as well as the influence of the microbiome on the metabolism of these drugs and their metabolites. Thursday @ 11:20 in Mozart 1-3 Clinical Validation of an Automated Dried Blood Spot Sampling Device – Fluispotter® Khem Adhikari - Rigshospitalet -- *Young Investigator Grantee* ‣ The use of dried blood spot (DBS) samples for the quantitative analysis has received increasing attention in the recent years. An accurate micro-volume spotting, 20-hours, automated and wearable DBS sampler, “Fluispotter®”, has been used and the analytical method for cortisol has been thoroughly validated as proof of concept. The method has good precision and accuracy and the device is well suited for clinical trials. Thursday @ 11:40 in Mozart 1-3 Use of Quantitative Metabolomics for Investigation of Age-Related Nuclear Cataracts Vadim Yanshole - International Tomography Center SB RAS -- *Young Investigator Grantee* ‣ A cataract (clouding of the lens) is the most common cause of vision declining of older people. Unlike most other human tissues, the lens has specific structure to be transparent: firstly, the lens consists mainly of fiber cells without organelles, and secondly, it lacks blood vessels. The protection of the lens is mainly provided by metabolites; most of them are synthesized in the lens epithelium or enter the lens through the epithelial layer from the surrounding aqueous humor (AH). Therefore, changes in the metabolome of the lens and AH may help to establish the molecular mechanisms of the cataract onset. Current report provides the data analysis of changes in the metabolomic profiles of human eye lens and AH under the development of cataracts. The concentration of more than 80 metabolites in the lens and AH have been determined with the combined use of LC-MS, LC-OD and NMR methods.

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Quantitation Issues Thursday @ 11:00 in Mozart 4-5

Session Chair: Christiane Auray-Blais - Universite de Sherbrooke Thursday @ 11:00 in Mozart 4-5 Semi-Quantitative LC/ESI/MS Analysis Using Predictive Models of ESI Ionization Efficiencies Jaanus Liigand - University of Tartu -- *Young Investigator Grantee* ‣ Until now, in the discovery of metabolites and in the absence of standard substances in LC/MS analyses equal ionization efficiencies are assumed. This may lead to misunderstandings of the processes occurring in organisms as concentrations of some metabolites can be up to 5 orders of magnitude over- or underestimated. By prediction of ionization efficiencies in both positive and negative electrospray ionization and in biological matrices the accuracy of such predictions can be improved, the best prediction being a 4-times mismatch with reality allowing for more accurate semi-quantitative analysis. This prediction method is user-friendly, as it uses 2D structures of the analytes and a small set of calibration compounds incorporated in the analytical run; thus, enabling quicker and more accurate estimation of the abundance of compounds of interest. Thursday @ 11:20 in Mozart 4-5 Quantification and Library Identification of 248 Compounds Commonly Detected in Systematic Toxicology Analysis Using an Automated Extraction Coupled to LC-MS/MS Robin Tiphaine - CHU of Limoges -- *Young Investigator Grantee* ‣ We report a fully automated extraction method carried out by a programable liquid handler directly coupled to an LC-MS/MS system for the identification and quantification of 248 compounds of interest. The acquisition was performed in positive and negative ionization mode with up to 15 MRM transitions per compound (MRM Spectrum mode). The method was validated according to the requirements of ISO 15189. A robustness study performed in 16 compounds demonstrated that samples could be quantified using a calibration curve dating up to one month. Thursday @ 11:40 in Mozart 4-5 Clinical Mass Spectrometry: the Assessment of Acidosis Profiles and Acute Kidney Injury in Severe Malaria Natthida Sriboonvorakul - Mahidol University -- *Young Investigator Grantee* ‣ In severe falciparum malaria acidosis and acute kidney injury (AKI) are independent predictors of a fatal outcome. The relationship between plasma acids, urine acids and renal function was investigated in adult patients with acute falciparum malaria. Clinical mass spectroscopy was utilized for assessment of small organic acid profiles. Plasma and urinary concentrations of selected acids were increased in falciparum malaria patients according to disease severity. Principal component analysis separated a group of patients with AKI and mainly driven by p-hydroxyphenyl lactate (pHPLA) concentrations in both plasma and urine. pHPLA could contribute to acute kidney injury in severe malaria.

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• Session 5 • Track 3 • Clinical Proteomics II

Thursday @ 11:00 in Papageno Hall Session Chair: Renee Ruhaak - Leiden University Medical Center

Thursday @ 11:00 in Papageno Hall Paving the Way for MS-based Protein Tests that Target Unmet Clinical Needs Christa Cobbaert - Leiden University Medical Center ‣ Proteins in body fluids are routinely tested in clinical laboratories by immunoassays, however diagnostic sensitivity- and specificity may not be sufficient for the intended use. Moreover, the applied immunoassays cannot recognize the different proteoforms, whereas MS-based strategies result in detailed structural characterization of specific protein targets. Furthermore, clinically effective tests that support patient management at an early and curable stage are lacking for a wide variety of diseases. In our institute we define specific unmet clinical needs with the clinicians according to a structured checklist developed by the European Federation of Laboratory Medicine Working Group on Test Evaluation and explore quantitative clinical chemistry proteomics as a strategy to detect and quantitate specific proteins in a multiplexed way. Thursday @ 11:20 in Papageno Hall Combination of Stable Labelled Antibodies and LC-MS to Accurately Quantify Therapeutic Monoclonal Antibodies in the Clinical Laboratory Ravindra Chaudhari - PROMISE Advanced Proteomics ‣ We will present an innovative and generic approach involving the use of stable isotope labeled mAbs used as internal standards and LC-MS to allow accurate, sensitive and specific quantification, as required for therapeutic drug monitoring. Our approach was already tested and implemented with success in pharmacology laboratories on anti-TNF antibodies (Infliximab and Adalimumab) and was recently implemented for monitoring Cetuximab. Thursday @ 11:40 in Papageno Hall Clinical Analytical Assay for AFP-L3 Using Multiple Reaction Monitoring-Mass Spectrometry for Diagnosing Hepatocellular Carcinoma Youngsoo Kim - Seoul National University College of Medicine ‣ We have developed and validated an MRM-MS assay for quantifying AFP-L3 in human serum to diagnose early-stage hepatocellular carcinoma. LiBA, the current standard method, cannot measure AFP-L3 concentrations accurately due to its low sensitivity. We addressed this issue with immunoprecipitation in conjunction with fractionation with LCA lectin. Consequently, the MRM-MS assay identified hepatocellular carcinoma patients that were missed by LiBA. In addition, we validated this approach in accordance with several multinational guidelines such as FDA, EMA, and CLSI. Our results demonstrated that this assay is robust and immediately applicable in clinical practice.

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Glycomics Keynote Thursday @ 11:00 in Paracelsus Hall

Session Chair: Manfred Wuhrer - Leiden University Medical Center Thursday @ 11:00 in Paracelsus Hall KEYNOTE PRESENTATION High-throughput Glycomics in Patient Stratification - What Did We Learn from the First 60,000 Analyses Gordon Lauc - University of Zagreb & Genos Glycoscience Research Laboratory, Zagreb, Croatia ‣ Since the onset of genome wide association studies, thousands of genetic loci have been associated with different diseases and traits. However, in the last few years it is becoming increasingly clear that variations in a DNA sequence are only a beginning of the understanding of complex human diseases. Genetic polymorphisms have to be put in the context of complex biology of life and a more elaborate approach that combines different ‘omics phenotypes is needed to understand disease mechanisms and perform patient stratification that transcends genomics. Glycomics, as by far the most complex epiproteomic modification, has an immense potential in this respect, which is only beginning to be investigated.

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Technological Innovations for the Microbiology Laboratory Thursday @ 11:00 in Trakl Hall

Session Chair: Ed Moore - University of Gothenburg Thursday @ 11:00 in Trakl Hall Laser Assisted Rapid Evaporative Ionisation Mass Spectrometry: An Automated Platform for Direct-From-Culture Speciation and Clinical Sample Analysis Simon Cameron - Imperial College London ‣ Rapid evaporative ionization mass spectrometry (REIMS) is a novel technique for clinical microbiology, and unlike commercially available MS platforms allows direct analysis of a culture plate or clinical diagnostic specimen. The automated and high-throughput REIMS platform previously developed now utilises a CO2 laser, rather than electrical current, for biomass heating; removing the requirement for contact to be made with the sample. This transition has allowed for an increase of over 50% in sample throughput and expands the range of sample types which can be analysed to include diagnostic specimens. This presentation will outline the current REIMS platform, the creation of a reference spectral database for over 60 microbial species, including bacteria, yeasts, and filamentous fungi, the early determination of antimicrobial susceptibilities, and direct from specimen pathogen detection. Thursday @ 11:20 in Trakl Hall In-situ Detection of the Botulinum Neurotoxin A by Functionalized MALDI Chips Petra Darebna - BioCeV ‣ Botulinum neurotoxin (BoNT) is well known for causing the lethal disease botulism. The only widely accepted test for the identification of BoNTs in both clinical specimens and food is the mouse bioassay. In this study, we use MALDI chips functionalized by biotin-binding protein neutravidin prepared by ambient soft ion landing at atmospheric pressure for enrichment and detection of biotinylated peptides – products of specific endoproteinase activity of BoNT/A. The functionality of surfaces modified with biotin-binding molecules could help to drive the MALDI-TOF mass spectrometry toward its implementation into routine clinical practice. Thursday @ 11:40 in Trakl Hall MALDI-TOF Mass Spectrometry: A Method for the Detection of Bacterial Contamination in Platelet Concentrates Yasmine Chetouane - IHU méditerranée infection -- *Young Investigator Grantee* ‣ Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been recently introduced in microbiological laboratories for the identification of microorganisms with proteomics approaches. Here, we investigate the use of MALDI-TOF MS by means of a Microflex MS (MALDI Biotyper-Bruker Daltonik, Germany) for the rapid, cheap and reliable detection and identification of bacterial contamination in Platelets Concentrates (PCs). We have demonstrated the stability and reproducibility of the spectrum over time and whatever the blood group of PCs. Also, we have compared the sensitivity and the specificity of the BACTEC cultivation method to the MALDI-TOF.

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Practical Training: Internal Standards Thursday @ 11:00 in Doppler Hall

Session Chair: Russ Grant - Lab Corp Thursday @ 11:00 in Doppler Hall Everything You Wanted to Know about Internal Standards But Were Too Afraid to Ask Russ Grant - Labcorp ‣ This one hour teaching session regarding Internal Standards will be delivered in three 20 minute vignettes. The first session will describe in detail the "What, Why and How" Internal standards should be used in isotope dilution LC-MS/MS assays. The second session "But what about when?" will describe situations where Internal Standards fail criterion for use and how to correct deficiencies for effective LC-MS/MS assays. The third session will describe the "Unique capabilities" afforded to analytical measurement when used as internal calibrators/pre-analytic correction tools and in method development (as surrogates for analytes).

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Keynote for Metabolomics Thursday @ 14:30 in Mozart 1-3

Session Chair: Russ Grant - Lab Corp Thursday @ 14:30 in Mozart 1-3 KEYNOTE PRESENTATION How Tandem Mass Spectrometry Revolutionized Newborn Screening David Millington - Duke University School of Medicine ‣ Inspired by a clinician’s account of a child rescued from near death by a revolutionary therapeutic intervention, the author applied chemistry and mass spectrometry to solve an analytical challenge that led to the first front-line diagnostic test performed by tandem mass spectrometry (MSMS) – the analysis of acylcarnitines to recognize and diagnose inherited disorders of fatty acid and branched-chain amino acid catabolism. By applying this method to dried blood spots and adding an additional analytical component to include several essential amino acids, a novel multiplex assay was developed to screen newborns for over 30 inherited metabolic conditions with a single test. The introduction of this method into public health systems and hospitals across the world during the past 20 years has literally revolutionized neonatal screening; new technology is being introduced to add more value.

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Monitoring Metabolites in Inborn Errors of Metabolism Thursday @ 14:30 in Mozart 4-5

Session Chair: Zdenek Spacil - Masaryk University Thursday @ 14:30 in Mozart 4-5 Development of a 2D-UPLC-MS/MS Assay for Therapeutic Monitoring for Patients with APRT Deficiency Unnur Arna Thorsteinsdottir - University of Iceland -- *Young Investigator Grantee* ‣ Adenine phosphoribosyltransferase (APRT) deficiency results in excessive urinary excretion of poorly soluble 2,8-dihydroxyadenine (DHA), causing nephrolithiasis and chronic kidney disease. Treatment with allopurinol and febuxostat effectively reduces DHA excretion and prevents urinary stone formation. However, a reliable method for therapeutic monitoring of patients with APRT deficiency is lacking. A 2D-UPLC-MS/MS assay for simultaneous quantification of the purines DHA, adenine, adenosine, inosine, hypoxanthine and xanthine and the pharmacological agents allopurinol, its active metabolite oxypurinol, and febuxostat, in human plasma samples will be developed and optimized utilizing design of experiments (DoE). To our knowledge, this is the first report of an absolute quantification of DHA in human plasma. Thursday @ 14:50 in Mozart 4-5 The Role of Chromatography in the Separation of Glucosylceramide Isoforms from Their Isobaric Galactosylceramide Counterparts in the Era of Precision Medicine Christiane Auray-Blais - Universite de Sherbrooke ‣ Glucosylceramide (GluCer) is a glycosphingolipid associated with various diseases, such as Parkinson’s disease and Gaucher disease. It is comprised of different fatty acid chains (isoforms), a sphingosine and a glucose moiety. The only difference between GluCer and galactosylceramide (GalCer) is the conformation of one hydroxyl group: axial in the former and equatorial in the latter. The focus of this study was to separate chromatographically GluCer isoforms from their isobaric GalCer counterparts. We report an ultra-performance liquid chromatography (UPLC) coupled to a tandem mass spectrometry (MS/MS) 6-min methodology which provides an efficient chromatographic separation of GluCer isoforms from their GalCer counterparts in brain tissues of Parkinson’s disease patients. This method was successfully applied to other matrices, such as vitreous humour in a Gaucher disease patient. Thursday @ 15:10 in Mozart 4-5 First Experience with a Second-Tier LC-MS/MS Assay for Newborn Screening of Propionic Acidemia, Methylmalonic Acidemias and Combined Remethylation Disorders Glynis Klinke - University of Heidelberg ‣ Increased propionylcarnitine levels in newborn screening are indicative for disorders such as propionic acidemia (PA), methylmalonic acidemia and combined remethylation disorders (MMACBL). Elevated propionylcarnitine occurs relatively frequently and requires a differential diagnosis. Thus, we aimed to develop a second-tier assay for concurrent determination of 3-hydroxypropionic acid, methylmalonic acid and methylcitric acid. The assay was developed using liquid chromatography coupled to tandem mass spectrometry, and clinically validated with retrospective analysis of DBS samples from PA or MMACBL patients. All three analytes were determined by this simple and fast assay, allowing a more specific and reliable identification of PA, MMACBL in stored newborn samples.

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Oncology Thursday @ 14:30 in Papageno Hall

Session Chair: Yuri van der Burgt Thursday @ 14:30 in Papageno Hall Proteomic Analysis of MGL Binding Protein in Human Cancer Cells Martina Pirro - Leiden University Medical Center LUMC -- *Young Investigator Grantee* ‣ O-glycosylation is generally initiated by the transfer of a N-acetylgalactosamine to Ser/Thr residues of proteins, forming the Tn antigen. This truncated surface glycan is expressed at high levels by tumor cells and is associated with higher metastatic behaviour and poor prognosis of patients. The Tn antigen is recognised by the C-type macrophage galactose lectin (MGL), which induces the activation of immunosuppressive responses. Here, we investigated the MGL binding proteins in Jurkat cells. The optimization of pull-down assays and subsequent glycoproteomic analysis by mass spectrometry, allowed us to identify 20 cell surface proteins as novel MGL-ligands. Thursday @ 14:50 in Papageno Hall Exosome-like Vesicles of Uterine Aspirates Permit the Identification of Diagnostic and Stratification Biomarkers of Endometrial Cancer Eva Colas - Vall Hebron Research Institute ‣ There is an urgent need to develop non-invasive tests that improve EC detection. In this study, we used exosome-like vesicles isolated from a uterine fluid to identify and verify protein signatures that can differentially diagnose EC subtypes. A discovery phase was performed using a super-SILAC approach on 60 patients (EC type 1, EC type 2, and controls), and a verification phase was done by targeted proteomics (SRM) in 107 patients. A 2-protein signature achieved an AUC=0.935 for EC diagnosis. In addition, we also report a new protein signature that can differentiate type1 versus type2 EC (AUC=0.932). This study has important implications in early detection of EC and in patient stratification. Thursday @ 15:10 in Papageno Hall In Depth Proteomic Analysis of Prostate Cancer Biopsies Gabor Toth - Hungarian Academy of Sciences -- *Young Investigator Grantee* ‣ Cancer research is among the most studied areas of science and prostate cancer (PCa) is one of the most common types of cancer among men. In this work we describe a detailed proteomics analysis of PCa tissue microarrays (TMAs) with our novel methodology. It is based on surface proteolytic digestion and proved to be capable of quantifying over 500 proteins from a single 1.5 mm diameter TMA core. We have compared the protein composition of tissues with various grades and stages of cancer. Samples from healthy and cancerous tissues were clearly distinguished and a good correlation with cancer grade was found. A well balanced study was carried out and over 100 proteins showed statistically significant abundance changes between various groups. During STRING evaluation up-regulation eg. in KEGG ribosome pathway and mRNA splicing could be observed.

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Tissue Imaging Keynote Thursday @ 14:30 in Paracelsus Hall

Session Chair: Kristina Schwamborn - TUM Thursday @ 14:30 in Paracelsus Hall KEYNOTE PRESENTATION Advances in Computational Methods for Tissue Imaging by Mass Spectrometry Raf Van de Plas - Delft University of Technology ‣ Imaging Mass Spectrometry (IMS) has made rapid progress as an imaging modality that can map the spatial distribution of molecules in tissue. In recent years, novel computational developments have become an increasingly important part of major advancements in this field. This talk presents several computational techniques developed in our group, specifically relevant to molecular imaging in medicine and the clinical practice. We show recent work in low-level signal processing, where in silico integration of isolation windows enables High-Dynamic-Range mass spectrometry, substantially increasing MS sensitivity. We also address advancements in data-driven image fusion, a multi-modal data mining methodology that drives the automated discovery of biomolecular relationships between stained microscopy and IMS, thus directly linking exploratory tissue analysis to established clinical targets.

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Biomarkers in Infectious Disease Thursday @ 14:30 in Trakl Hall

Session Chair: Stefan Zimmerman - University Hospital Heidelberg Thursday @ 14:30 in Trakl Hall Sepsis by Enterobacteria: A MALDI Affair Markus Kostrzewa - Bruker Daltonik GmbH ‣ Enterobacteria are the most frequent causative agents of sepsis. The spread of cephalosporinase- and carbapenemase-production in these species is a worrying threat for the effectiveness of the antibiotic therapy. Every hour saved in the detection of such strains can be crucial for the patients’ clinical outcome. In this study, we investigate an innovative full MALDI-TOF MS based approach to quickly detect cephalosporinase- and carbapenemase-producing enterobacteria directly from the positive blood cultures bottles, applying the novel tools of the Biotyper system (Bruker Daltonik). The bacterial pellet extracted by Sepsityper was used for the species identification, for the detection of KPC-producing strains by subtyping, and for evaluation of cephalosporinase and carbapenemase activity by STAR-Cepha and STAR-Carba hydrolysis assays. Thursday @ 14:50 in Trakl Hall Omics Approaches Towards Discovery of Biomarkers for Early Diagnosis of Tuberculosis Ana Varela Coelho - ITQB NOVA -- *Young Investigator Grantee* ‣ Tuberculosis (TB) is a disease with worldwide presence and a major cause of death in several developing countries. Current diagnostic methodologies often lack specificity and sensitivity, whereas in some cases it takes a long time to obtain a conclusive result. Four metabolites and four proteins were selected as potential biomarkers of TB infection using NMR based metabolomics combined with differential proteomics. These biomarkers are strong candidates for the development of a clinical test. Thursday @ 15:10 in Trakl Hall Identification and Validation of Two Peptide Markers for the Recognition of Clostridium difficile MLST-1 and MLST-11 by MALDI-MS Simone Nicolardi - Leiden University Medical Center ‣ Clostridioides difficile has rapidly emerged as the main cause of antibiotic-associated diarrhoea as a consequence of colon inflammation. C. difficile infections-associated mortality can be high and spread can occur, leading to nosocomial outbreaks. Consequently, there is a need for rapid tests which not only identify the infections, but also provides insight into the relatedness of the strains. In this study, we used ultrahigh resolution MALDI-FTICR MS to generate detailed and comprehensive bacterial protein profiles and identify specific peptide markers for MLST-1 and MLST-11. The elucidation of the amino acid sequence of these markers allowed the investigation of a large publically available genome database of C. difficile MLSTs and the accurate determination of their sensitivity and specificity. A method was developed to identify the MLST-1 marker also by low resolution MALDI-TOF MS.

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Practical Training : Getting to Lower Limits of Quantitation Thursday @ 14:30 in Doppler Hall

Session Chair: Brian Rappold - Lab Corp Thursday @ 14:30 in Doppler Hall 1/3 Getting Lower Limits of Quantification in LC-MS/MS assays : Prologue Brian Rappold - LabCorp ‣ Understanding mechanisms to achieving better sensitivity in LC-MS/MS assays must start with fundamentals and first principles of the technology. An initial conversation of noise, response functions, ionization techniques and hardware will introduce attendees to the basics of mechanisms used to reduced needed sample volume, increase instrument response and improve assay precision. Thursday @ 14:50 in Doppler Hall 2/3 Getting Lower Limits of Quantification in LC-MS/MS assays : Phases and Stages Brian Rappold - LabCorp ‣ The second of 3 lectures on improving instrument response will focus on the approaches and experimentation used in LC-MS/MS assays. Understanding that increased response can only be generated in two phases of the LC-MS/MS process (material loaded into the system and ionization cross-section), a series of examples on scientifically defining “optimization” will be walked through. Thursday @ 15:10 in Doppler Hall 3/3 Getting Lower Limits of Quantification in LC-MS/MS assays : The Words Don’t Fit the Picture Brian Rappold - LabCorp ‣ In the concluding lecture of the series, data from experiments previously described will be shown, including the results of precision-improving transition summing and solvent screening. The rationales behind analyte losses in handling and analysis will be discussed, as well as the limitations to the approaches examined. Understanding why titles of albums by Willie Nelson were selected for lecture titles shall also be briefly investigated.

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2019 US Ad

77

Poster Presentations

Grouped by Topic

Location: Exhibit Hall (1st Floor)

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Endocrinology : Poster Presentations

Poster #6a in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Endocrinology Development and Validation of the Simultaneous Measurement of Estrone and 17β-Estradiol in Serum by LC-MS/MS for Clinical Laboratory Applications Neus Fabregat-Cabello - University of Liège, CHU de Liège *YI Grantee* ‣ A straightforward analytical method for the determination of estrone (E1) and 17β-estradiol (E2) at ultra-low levels in serum samples by LC-MS/MS has been developed and validated. This method entails an extraction and derivatization with dansyl chloride followed by separation with a C18 column. A comparison of the developed LC-MS/MS method against our routine immunoassay shows a good correlation for E2 while an important negative bias is observed for E1 by RIA. The later comparison for E1 shows the need to switch from the current routine automated immunoassays to highly-sensitive LC-MS/MS quantifications in order to provide accurate and reliable clinical results, especially at very low levels.

Poster #6b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Endocrinology Determination of Cortisol and Cortisone in Urine by LC-MS/MS: Impact on Quality Results and Cost Benefits Matteo Vidali - Maggiore della Carità Hospital ‣ The 24-h urinary free cortisol measurement is widely used as first laboratory approach in the diagnosis of Cushing syndrome. Several immunometric methods are available; however, main concerns are well known and include cross reactivity with similar analytes, lack of standardization between labs and personnel costs due to manual sample preparation. We developed a simple and selective method for the analysis of urinary cortisone and cortisol by LC-MS/MS suitable for use in a clinical laboratory routine. Immunometric and LC-MS/MS methods were compared with regard to analytical results and costs.

Poster #7b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Endocrinology Evaluation of an IVD-CE Certified Plasma Metanephrine Assay: Lessons Learned from External Comparison Data Valentin Braun - University of Innsbruck ‣ We evaluated the performance and suitability of a commercial IVD-CE certified assay for LC-MS/MS analysis of free metanephrines in plasma for the use in routine clinical screening of catecholamine secreting tumours. The assay delivered good results on our analytical platform in terms of sensitivity (limit of quantification), internal QC precision (< 10.8 % at the lowest QC-level) and internal QC accuracy (< −5.8 %). However, the assay showed a constant positive bias for all three analytes in comparison to external quality control scheme data (mean < +4.9 %) and in a laboratory comparison experiment (mean < +11.8 %), exceeding the internal QC bias. The effect can be considered clinically insignificant but certainly deserves attention in the future, e.g. by striving for measurement traceability on an international level.

Poster #7c in Exhibit Hall - attended for 1 hr on Thursday starting at 10:00 Topic: Endocrinology Evaluation of Sample Preparation Options for the Simultaneous Extraction of Angiotensin and Aldosterone Prior to LC-MS/MS Analysis Ryu Konoshita - Shimadzu Europa GmbH ‣ This poster compares options for the simultaneous extraction of aldosterone and angiotensin from plasma. LC-MS/MS analysis was performed using a Shimadzu Nexera UHPLC system coupled to an 8060 triple quadrupole MS. Sample preparation strategies compared polymer-based reverse phase and mixed-mode sorbent chemistries. Analyte properties and solubilities were used to great effect resulting in non-compromised extract cleanliness for each fraction. Analyte recoveries greater than 75% and good removal of phospholipid interferences were demonstrated by the final protocols. Calibration curves demonstrated excellent linearity and corresponding r2 values greater than 0.99. Full results, discussion and conclusion will be shown in the final poster.

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Poster #10a in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Endocrinology Development and Validation of a LC-MS/MS Assay for Simultaneous Analysis of Salivary Cortisol and Cortisone Charlotte Harborow - Liverpool Clinical Laboratories *YI Grantee* ‣ A method has been successfully developed and validated for the simultaneous analysis of salivary cortisol and cortisone using a liquid phase extraction prior to LC-MS/MS analysis. Full validation of the method was performed according to published FDA and CLSI criteria. Chromsystems matrix-matched calibrators were evaluated; use of these calibrators has not been described previously. Interference from other synthetic and endogenous steroids was assessed, including several inhaled steroids. This assay can be used as a first-line test for patients presenting with clinical features of Cushing’s syndrome; providing a simpler, less-invasive sampling technique than other available diagnostic tests.

Poster #13b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Endocrinology Comparison of Atmospheric Pressure Chemical Ionization (APCI) and Electrospray ionization (ESI) and Matrix Types for Determination of Serum Steroid Hormones Atecla Alves - Universidade de São Paulo HCFMUSP ‣ Our aim was to compare equivalence of APCI and ESI ionization and two types of matrices to prepare calibrators for simultaneous analyses of five serum steroids hormones with LC/MS-MS. We selected 27 random samples to be measured by APCI or ESI with standards prepared in hormone-free pooled serum or in 5% BSA. Comparing APCI and ESI, androstenedione, and 21-deoxycortisol presented an inadequate index of total allowable error (TEA) for ESI. Cortisol, 11-deoxycortisol, and 17-OHP showed acceptable TEA for the four methods tested. We concluded that 5% BSA is an adequate matrix and that APCI is more versatile than the ESI method.

Poster #14b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Endocrinology Steroid Panel In Serum Based On LC-MS/MS Marcel Borren van - rijnstate Hospital ‣ We developed a 17-steroid panel in serum based on isotope dilution LC-MSMS. Full chromatographic separation was obtained within 15 minutes for all steroids on a biphenyl column using a slowly increasing methanol gradient. Addition of ammonium fluoride increased ionization for most steroids but not for all.Recovery of steroids proved best with supported liquid extraction. Therefore we concluded that this application is not ready for thorough validation.

Poster #17c in Exhibit Hall - attended for 1 hr on Thursday starting at 10:00 Topic: Endocrinology Performance Characteristics of the New Fully-Automated LC-MS/MS Total Vitamin D Assay on the CascadionTM SM Clinical Analyzer Marta Kozak - Thermo Fisher Scientific ‣ The Cascadion Analyzer is an easy to use, random-access analytical platform implementing LC-MS/MS technology. Performance of the Vitamin D assay for the quantitation of 25-hydroxy Vitamin D3 and 25-hydroxy Vitamin D2 in human serum and plasma was evaluated according to CLSI guidelines. NIST standard reference materials and CDC certified serum samples were analyzed to evaluate accuracy. Precision, accuracy, linearity, limit of quantitation, absence epimer interference, and method robustness met accepted laboratory criteria. Sixteen types of collection tubes were qualified. A calibration curve stability of 2 weeks was demonstrated. A reference range was established using the US population.

Poster #17d in Exhibit Hall - attended for 1 hr on Thursday starting at 12:30 Topic: Endocrinology High Sensitivity Assay of Estrogens in Human Plasma by UHPLC-MS/MS without Derivatization Neil Loftus - Shimadzu Corporation, MS Business Unit Overseas ‣ While many laboratories still use immunoassay to measure estrogen levels in plasma, the se assay suffer from several drawbacks and are not able to measure very low levels in routine. Here we present an UHPLC-MS/MS method that can measure low pg/mL of estrone, estradiol and estriol with an easy and rapid sample preparation. Limits of quantification were of 0.5, 1 and 5 pg/mL for E1, E2 and E3, respectively, using a sample volume of 200μL. The total analysis time was of 3.5 min. Validation experiments and real samples have been assayed to demonstrate the fit-for-purpose of the method.

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Poster #17i in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Endocrinology Salivary Cortisol and Cortisone by LC-MS/MS in a ISO15189 Accredited Laboratory: What Happens when Changing the Calibrators? Giorgia Antonelli - UNIVERSITY OF PADOVA ‣ The examination procedure (EP) salivary cortisol (sF) is accredited according to the ISO15189:2012 since 2016. This EP is a laboratory developed method, using home-made calibrators; according to the requirement 5.5.1.3, this method was validated, producing a validation certificate. Recently CE-IVD calibrators were available and our aim was to substitute the home-made calibrators with these ones. Comparing the obtained results, a significant bias was obtained for the salivary cortisol and cortisone ratio (sFEr), so the reference intervals (RI) were modified according to the Passing Bablok regression. The new RI had to be verified, according to CLSI EP28. Twenty healthy volunteers were recruited and saliva was collected. The results were suitable and a new validation certificate was produced.

Poster #22i in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Endocrinology Evaluation of the Analytical Performance of a LC-HR-MS Method for the Quantification of Hepcidin-25 in Human Serum Sieglinde Zelzer - Clinical Institute of Medical and Chemical Laborat ‣ We aimed to establish a valid and reliable hepcidin-25 LC-HR-MS assay for clinical routine. After method implementation, a study cohort of 200 serum samples was investigated. Evaluated data were within the (±15%) limits of usual validation guidelines, with a limitation in freeze thaw experiments. Freeze-/thaw-cycle experiments showed no significant differences (8%) for two but significant higher levels for three freeze-/-thaw cycles (20%). Females had significantly lower hepcidin-25 concentrations (median 8.42; range 3.71 – 16.81 ng/ml) compared to males (median 15.76; range 8.03 – 31.21 ng/ml; p = 0.004). To increase the precision of this reliable and robust method we propose performing a multi-point calibration for hepcidin-25 measurements.

Poster #23a in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Endocrinology Simultaneous Quantification of Plasma 3-Methoxytyramine, Metanephrine and Normetanephrine by Ultraperformance LC-MSMS Erdim Sertoglu - Gulhane School of Medicine ‣ Metanephrine (MN), normetanephrine (NMN), and 3-methoxytyramine (MTY) are are produced by O-methylation of the catecholamines. In this study, we aimed to develop a rapid and sensitive mass spectrometry based method coupled to ultraperformance liquid chromatography (UPLC-MS/MS) to measure these plasma catecholamine metabolites for the diagnosis of neuroendocrine tumors. Reversed-phase HPLC separation was performed using a Raptor HILIC-Si LC column (50 x 2.1 mm (i.d.); 2.7 μm particle size) after extraction onto Oasis WCX (1 mL, 10 mg) 30 µm solid-phase extraction cartridges. This method, with good precision, sensitivity and linearity, can be used in clinical and research laboratories.

Poster #23e in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Endocrinology Simultaneous Determination of Ten Different Steroid Hormones Using Ultra Performance Liquid Chromatography -Tandem Mass Spectrometry Method Ercan Saruhan - Mugla Sitki Kocman University, School of Medicine ‣ Due to having limitations on measurement by immunoassays, many mass spectrometry based studies have been conducted to ensure accurate measurement of steroid hormones so far. We aimed to develop a rapid, sensitive and high-throughput UPLC-MS/MS method for the simultaneous quantification of 10 steroids. After liquid-liquid extraction by methyl tert-butyl ether, the chromatographic separation was carried out on an Accucore C18 2.6µm, 100mm x 2.1mm column with a gradient mobile phase consisting of 0.2mM ammonium fluoride in water and 100% methanol at a flow rate of 0.5 mL-min⁻¹. This method, with good precision,sensitivity and linearity, can be used in clinical laboratories.

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Glycomics : Poster Presentations

Poster #3c in Exhibit Hall - attended for 1 hr on Thursday starting at 10:00 Topic: Glycomics An MS-based Middle-Up Approach for the Allotype-Specific Analysis of Human Plasma IgG Fc N-Glycosylation Thomas Senard - Leiden University Medical Center *YI Grantee* ‣ Human plasma immunoglobulins G (IgG) are a complex mixture of proteins with several posttranslational modifications (PTM), which are known to modulate antibody effector functions. We here propose a new middle-up strategy for the analysis of the fragment crystallizable (Fc) portion of polyclonal IgG, with a focus on specific glycosylation and PTMs of IgG allotypes. The analyses were performed by hydrophilic interaction liquid chromatography (HILIC) and capillary electrophoresis (CE) coupled with mass spectrometry (MS). These two orthogonal systems allow the resolution of the Fc allotypes with their different proteoforms, such as glycosylated, oxidized, C-terminal lysine-clipped or deamidated IgGs.

Metabolomics : Poster Presentations

Poster #1c in Exhibit Hall - attended for 1 hr on Thursday starting at 10:00 Topic: Metabolomics Profiling Pathways of Estrogen Metabolism in Patients with Pulmonary Arterial Hypertension: Development of a Novel Assay for Estrogen Metabolites by LC-MS/MS Nina Denver - University of Glasgow *YI Grantee* ‣ Pulmonary arterial hypertension (PAH) is a debilitating, life limiting, commonly misdiagnosed disease, which ultimately leads to right heart failure and death. Estrogen metabolism plays a key role in disease onset and progression. To simultaneously quantify circulating estrogen levels in human plasma a sensitive liquid chromatography mass spectrometry (LC-MS/MS) was developed to compare plasma from healthy controls and PAH patients. Initial studies show increases in metabolites involved in PAH pathophysiology prompting further investigation for potential diagnostic and therapeutic strategies. Furthermore, implementation of this method will be clinically relevant to a number of estrogen sensitive diseases.

Poster #2d in Exhibit Hall - attended for 1 hr on Thursday starting at 12:30 Topic: Metabolomics Lipidome Profile of Pathology Endometrium by Tandem Liquid Chromatography-Mass Spectrometry Alisa Tokareva - Moscow Institute of Physics and Technology ‣ Myoma and endometriosis are the most widespread pathology of endometrium. Understanding of changing of metabolism. 40 samples of proliferative or secretor endometrium with myoma or endometriosis were analyzed by tandem liquid chromatography-mass spectrometry. Chromatograms were deconvoluted by MzMine 3.0, lipids were identified by LipidMatch scripts. Lipids with statistical significant difference in levels for myoma and endometriosis of proliferative endometrium and for myoma and endometriosis of secretor endometrium. Levels of phosphotidylethanolamines, ceramides and sphingomyelines increases in myoma’s cell on proliferative endometrium phase. Levels of phosphotidylcholines with stearic fatty acid decreases in myoma of secretor endometrium phase.

Poster #3b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Metabolomics Intact-cell Mass Spectrometry for Pluripotent Stem Cell Identification and Authentication Petr Vanhara - St. Anne's University Hospital Brno ‣ Human embryonic stem cells (hESCs) emerged as promising tool in cell therapy, bio-industry or drug development. However, long-term cultured hESCs develop hidden phenotypic changes, preventing clinically relevant use. Quality control based on karyotyping, (epi)genetic traits, or monitoring of adverse phenotypic changes often suffers of low sensitivity. Therefore, there is ongoing need for methods revealing abnormalities in cell phenotype. We modeled shifts in hESCs using different strategies and addressed changes in cell status by intact cell mass spectrometry. Analysis of spectral fingerprints by chemometrics and machine-learning methods distinguished otherwise identical hESCs and provided efficient tool for quality control of hESCs cultures.

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Poster #4b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Metabolomics Separation and Quantitation of Acyl-Carnitines and Acyl-Glycines Bernhard Schoenenberger - Sigma-Aldrich Production GmbH (Merck KGaA) ‣ The influence of several analytical parameters of acyl-carnitines and acyl-glycines were studied. Those results were applied for the development of chromatographic separation of multi component mixtures of these products by HPLC-MS and -CAD towards the development of ready-to-use certified reference material mixtures.

Poster #5c in Exhibit Hall - attended for 1 hr on Thursday starting at 10:00 Topic: Metabolomics The Development of Steroid Panel for Mass Spectrometric Diagnosis of Polycystic Ovary Syndrome Natalia Kitsilovskaya - National Research Center of Obstetrics and Gynecology, Russia *YI Grantee* ‣ There are several phenotypesofPolycystic ovary syndrome (PCOS) and precise determination of steroid hormone levels is essential for adequate PCOS diagnostics ant therapy. Widely used immunoassays are not completely reliable especially in detection of low concentrations of hormones. Liquid chromatography with triple quadrupole mass spectrometry is more sensitive, specific and economically effective, allowing simultaneous analysis of steroid panels with numerous hormones. The present study deals with development of steroid hormones panel for PCOS diagnostics and its validation on a group of patients. Key words: polycystic ovary syndrome, steroid hormones, quantitative analysis, triple quadrupole.

Poster #5d in Exhibit Hall - attended for 1 hr on Thursday starting at 12:30 Topic: Metabolomics False Positive Signals in Profiling of Vitamin D Metabolites by LC-MS/MS Rafal Rola - Nicolaus Copernicus University ‣ One of the critical parameters describing method for determination of vitamin D metabolites is the ability to separate all compounds, since many of them have similar structures. Preliminary experimental data indicate the presence of number of different metabolites, especially double hydroxylated ones. One of them is 25,26(OH)2D3 , which interferes with 24,25(OH)2D3 catabolite. This endogenous compound was determined in serum of idiopathic infantile hypercalcemia (IIH) patients with confirmed knockout of 24-hydroxylase (CYP24A1) gene. Further studies shown presence of 25,26(OH)3D3 also in serum of healthy individuals, which affects significantly on 24,25(OH)2D3 results.

Poster #8a in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Metabolomics Lipidomics in Breast Cancer Cell Lines: A Way to Distinguish Breast Cancer Subtypes by Lipid Composition? Martha Kampp Nøhr Vestergaard - University of Iceland *YI Grantee* ‣ Early diagnosis of breast cancer (BC) is a prerequisite for early treatment onset to improve prospects for the patient. The treatment of BC is selected depending on the BC subtype. This study aims to identify novel lipid biomarkers for distinguishing BC subtypes and for early diagnosis of BC. The lipid composition of six BC cell lines (MDA-MB-231, MDA-MB-436, SK-BR-3, T-47D, MCF7 and CAMA-1) were analysed using UPLC-QToF-MS. The cell lines represent different subtypes of BC. Multivariate data analysis of detected lipid markers showed separated clusters of the individual BC cell lines and indicate different lipid composition between the BC subtypes. Lipid markers from the lipid classes PC, PE, LPC, LPE, SM and TG were identified. Further data interpretation will identify novel lipid biomarkers for BC subtypes and aid early diagnosis of BC from clinical samples.

Poster #8b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Metabolomics A HILIC-ESI-MS Method for Profiling Biomarkers of Maple Syrup Urine Disease and Methyl Malonic Acidemia in Urine Elizabeth Mathew - MANIPAL COLLEGE OF PHARMACEUTICAL SCIENCES *YI Grantee* ‣ Methyl malonic acid and branched chain keto acids are important biomarkers for the diagnosis of cobalamin deficiencies and maple syrup urine disease. Urine samples were 100 times diluted and analysed on a ZIC-HILIC column with 25µM formic acid in water: 25µM formic acid in acetonitrile (45:55) at a flow rate of 0.8 mL/min within 6 minutes. The method demonstrated a sensitivity of 50 ng/mL and recoveries of 87-105% with good precision. Stability studies demonstrated a percent change of ±15% in stock and matrix. The Bland Altman analysis of the developed method with GC-MS method demonstrated a bias between 0.009-0.44 for all analytes.

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Poster #9a in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Metabolomics The Oxytocin Study: A Translational Approach from Molecular Biology to Behavior Analysis Laura Albantakis - Max Planck Institute of Psychiatry *YI Grantee* ‣ The main objectives of the study are to investigate whether the molecular sets (including Oxytocin, BDNF and metabolomics) of ASD patients are altered compared to those of healthy controls and depressed patients. And if so, which analytes reflect the autistic phenotype the most accurately? In this context confounding factors (such as age, hormonal status, comorbidities, medication etc.) and chosen methods (biomaterial, analysis via EIA, LC-MS or RIA) will be closely explored and validated. In a second step, the results of the molecular analysis will be related to data from imaging and behavioral studies. Based on these objective and observer-independent findings, an autistic phenotype will be designed, which will further elucidate the etiology of ASD, and highlight therapeutic approaches.

Poster #9b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Metabolomics Development of an Analytical Method for Serum P-Cresylsulphate and Indoxylsulphate in Patients with Chronic Kidney Disease Marco Bagnati Giorgio Bellomo - Maggiore della Carità Hospital ‣ P-Cresylsulphate (PCS) and Indoxylsulphate (IXS) have been previously found to be increased in subjects with end stage renal disease (ESRD) and associated with cardiovascular mortality in chronic kidney disease (CKD). We developed a LC-MS/MS method for measuring serum PCS and IXS in patients with CKD. We found significantly higher total (protein-bound and unbound) and free (unbound) median PCS and IXS levels than healthy subjects. Higher levels were associated with higher stages of CKD. Our results confirm that free and total PCS and IXS levels are increased in CKD patients possibly be regarded as valid markers for the progression of CKD.

Poster #10b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Metabolomics A Novel Signalling Role for Bacteria Present in Gastro-Oesophageal Tumor Microenvironment and its Contribution to Compound Production Using Mass Spectrometry Mina Adam - Imperial College London UK *YI Grantee* ‣ Cancer of the stomach and oesophagus is among the world's top five cancers. Survival rates are very poor as the disease presents late and early symptoms are non-specific. Our group is currently developing a non-invasive test for cancers of the stomach based on the detection of small molecules in exhaled breath that we believe are produced by the cancer as well as the associated bacteria. The potential role of bacteria and their influence on Volatile Organic Compounds (VOCs) produced within the upper gastrointestinal tract is currently unknown. In this study, Gas Chromatography Mass Spectrometry has been utilised for the identification of VOCs from specific bacteria, which enhanced the accuracy of the breath test.

Poster #10c in Exhibit Hall - attended for 1 hr on Thursday starting at 10:00 Topic: Metabolomics First Very Long-Chain Acyl-CoA Dehydrogenase Deficiency (VLCADD) Patient Detected Through Newborn Screening in Croatia Ivana Križi? - University Hospital Center Zagreb ‣ Expanded newborn screening (NBS) pilot project in Croatia using tandem mass spectrometry (LC-MS/MS) has started in October 2017. In this abstract we describe the first case of very long-chain acyl-CoA dehydrogenase deficiency (VLCADD), one of six additional diseases included, detected through NBS in Croatia. An asymptomatic male newborn was highly suspicious for VLCADD based on NBS results. Levels of tetradecenoylcarnitine as a primary VLCADD marker were elevated, as well as levels of secondary markers. Urinary organic acids showed slightly elevated concentrations of several dicarboxylic acids and plasma acylcarnitine profile revealed increased concentration of C14:1. VLCADD diagnosis was confirmed by genetic analysis of ACADVL gene.

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Poster #10d in Exhibit Hall - attended for 1 hr on Thursday starting at 12:30 Topic: Metabolomics Investigation of Reference Ranges of Long-Chain Fatty Acids in Pediatric Population Ilgar Mamedov - LLC ChromsystemsLab, Moscow, Russia ‣ Long chain fatty acids, as well as the fatty acid with a branched chain (pristanic and phytanic acids), are extremely hydrophobic and practically insoluble in water. Patients with abnormal peroxisome function can be observed, the increased content of long chain fatty acids (with chain greater than 26 carbon atoms) can be determined. Hexacosanoic acid C26:0, lignoceric acid C24:0, behenic acid C22:0, and their relations can also be used for diagnostics. Determination of long chain fatty acids content was performed by gas chromatography with mass spectrometric detection (GC-MS).

Poster #11a in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Metabolomics Characterization of Clinically Relevant Markers of Microbial Colonization and Alzheimer’s Disease Zdenek Spacil - Masaryk University *YI Grantee* ‣ Human gut microbiota is arguably the most influential factor affecting health. Bacterial metabolic pathways utilize aromatic amino acids and particularly their interaction with tryptophan metabolism causes lifelong immunomodulation in human. However, specific molecular mechanisms of shaping immune response, distribution of microbiota-modulated metabolites in biofluids or corresponding levels of inflammation markers are poorly understood. We applied mass spectrometry based omics technologies to profile a panel of tryptophan metabolites in several biological matrices, to develop a multiplex assay for quantification of inflammatory proteins and to characterize membrane lipids involved in the immune response to pathogens. We applied the systems approach to investigate microbial influence on Alzheimer’s disease (AD) in effort to identify perspective clinical markers.

Poster #14c in Exhibit Hall - attended for 1 hr on Thursday starting at 10:00 Topic: Metabolomics Obesity-Driven Microglial Response in the Murine Brain - Functional Role of Cerebral Lipid Metabolism Madlen Reinicke - University Leipzig *YI Grantee* ‣ In mice brain after 24 weeks of feeding, detailed picture of the microglial response to obesity were achieved by immunostaining. To identify lipid biomarker candidates with significant diet associated differences in brain of mice on a high-fat diet compared to normal diet, targeted LC-MS/MS analysis of more than 270 analytes related to the lipid metabolism including cholesterol and bile acid metabolism, apolipoproteins, amino acids, acyl carnitines, eicosanoids and resolvins were performed. Correlation of microglial activation and effected lipid pathways were studied.

Poster #15f in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Metabolomics Metabolic Profiling of ADPKD Patient Urine Using NMR Spectroscopy Shosha Dekker - Leiden University Medical Center *YI Grantee* ‣ Quantitative NMR profiling enabled identification of metabolic markers that distinguished ADPKD patients from healthy controls. The ratio of the two most promising metabolites associates well with eGFR in a cohort of patients with ADPKD.

Poster #17g in Exhibit Hall - attended for 1 hr on Thursday starting at 10:00 Topic: Metabolomics Skin Biomarkers for Cystic Fibrosis Screening Jeany Delafiori - University of Campinas (UNICAMP) *YI Grantee* ‣ Genetic disorders such as cystic fibrosis are conditions that lay a heavy burden over the individuals affected by it. The early detection of the disease may not always be readily performed, as many variables are involved in its accurate diagnosis, directly affecting the quality of life of the individuals. Using High Resolution Mass Spectrometry as the analytical tool and sample collection based on skin imprint in silica plates, 8 biomarkers related to the cystic fibrosis pathophysiological condition were identified. These biomarkers showed great potential for diagnostic purposes and monitoring of disease evolution.

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Poster #19b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Metabolomics GC-MS Targeted Screening of Different Metabolite Classes for Prostate Cancer and Prostate Hyperplasia Patients and Correlation with PSA Pavel Markin - Sechenov First Moscow State Medical University *YI Grantee* ‣ Metabolite profiling techniques, such as GC-MS, are finding increasing applications in the diagnosis of prostate cancer. For biofluid analysis, three different methods for reaching fatty acids by their methyl esters, amino acids by methylchloroformiate detivatives and organic acids and sugars by MSTFA derivatives were made and tested using GC-MS (Agilent 7820A Series GC/MSD). 38 men with benign prostate hyperplasia and 27 men with malignant prostate cancer were analyzed. 23 metabolites showed significant difference between groups (p<0.05). Spearman r-test was performed in searching for correlation between metabolites and PSA (prostate specific antigen) levels. 9 metabolites were strongly correllated (p<0.05).

Poster #21a in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Metabolomics Comparative Analysis of Lipid Composition of Endometrium Between Patients with Endometriosis and Uterine Myoma Dinara Salimova - Kulakov National Medical Research Center of ObGYN *YI Grantee* ‣ There is luck of reliable minimally or non-invasive diagnostic methods of endometriosis and insufficient knowledge of its pathophysiology. Previous studies demonstrated differences in lipid metabolism comparing eutopic and ectopic endometrium. The current investigation deals with differences in lipid composition of eutopic endometrium among patients with endometriosis and uterine fibroids. Statistically significant variation of some lipid species belonging to triacylglycerol and phosphatidylcholine and cardiolipin classes between groups was found.

Poster #22c in Exhibit Hall - attended for 1 hr on Thursday starting at 10:00 Topic: Metabolomics Determination of Combretastatin A4 Influence on Human Lung Cancer and Mouse Melanoma as in vitro Cell Culture Models by SPME-LC-HRMS Karol Jaroch - Dep. of Pharmacodynamics and Molecular Pharmacolog *YI Grantee* ‣ Use of solid phase microextraction (SPME) in combination with high-resolution mass spectrometry (HRMS) for cell culture metabolomics analysis allows getting more sophisticated data from in vitro assays. SPME was employed for determination of metabolomic changes after induction of non-small cell lung cancer cell line (A549) or melanoma (B16F10) cell lines with combretastatin A4 phosphate and combretastatin A4. Both, up- and downregulation of aminoacids (e.g.: valine, proline,) low molecular mass acids (pyroglutamic acid) and amides (palmitamide) was found as distinguishing between treated and non-treated cells. The whole procedure can be considered as HTS and „non-invasive” for cells. The use of SPME-(LC)-HRMS with cell cultures in high-throughput manner were presented for the first time. Work has been financed by Nicolaus Copernicus University statutory grant No 451.

Poster #23f in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Metabolomics Comparison of Two LC-MS-based Screening Methods for Acyl-Carnitines in Fatty Acid Oxidation and Abnormalities in LCHAD Deficiency Günter Fauler - Medical University of Graz ‣ Carnitine profiling in plasma (free carnitine and acyl-carnitines, ACs) is a useful method in newborn screening for diagnosis of fatty acid oxidation defects. The aim of this work was to compare a new UHPLC-high resolution-mass spectrometry method (analysis of butyl esters by ESI and Q-Exactive Orbitrap in full scan mode) with a method using HILIC-chromatography and 4000QTrap system (without derivatization) in MRM mode. Both methods are suitable for routine measurements. In samples of affected children an abnormality was detected in patients with a LCHAD deficiency: isobaric double peaks appeared in the chromatograms of particular acyl-carnitines. Attempts have been done to get insight in this characteristic form of appearance.

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Poster #24b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Metabolomics Multidirectional Differential Metabolomic Analysis of Selected Brain Tumors in Humans Paulina Zofia Goryńska - Faculty of Pharmacy, Collegium Medicum *YI Grantee* ‣ Fast and reliable diagnostic methods are still the point of interests for clinicians. Some of the diseases e.g. cancers do not cause any early symptoms. The main goal for the technological progress in terms of diagnostic methods is the development of alternative tools complementary to currently used techniques thus providing a full panel of results for the patient. In current study solid phase microextraction (SPME) has been successfully used for untargeted tissue analysis as a method applicable for in vivo and in situ analysis with no requirement for tissue collection.

Microbiology : Poster Presentations

Poster #3a in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Microbiology Comparative MALDI-TOF Analysis of Staphylococcus spp. from the Environment including Antibiotic Resistant Strains Rory Cave - University of East London *YI Grantee* ‣ In this study, we did a comparative analysis of the Bruker’s Autoflex with ASTA Tinkerbell MALDI-TOF MS using samples spotted onto the same target plate. We used 10 references strains from different species to determine how accurate they identify the species. We also tested a range of environmental staphylococcal isolates that were resistant to antibiotics. From the result, we were able to determine that both excellent congruences between data analysed using ASTA’s Tinkerbell and Bruker’s Biotyper at the species and subspecies levels.

Poster #16c in Exhibit Hall - attended for 1 hr on Thursday starting at 10:00 Topic: Microbiology Detection of Resistance Markers by MALDI-TOF MS – Yesterday, Today, Tomorrow Miriam Cordovana - Univ. Hospital of Bologna Sant'Orsola-Malpighi ‣ Bacterial resistance to antibiotics represents a global threat. MALDI-TOF MS technology proved to be a reliable and robust method for real time detection of specific resistance markers in the bacterial mass spectrum during the species identification process. Here, we investigated the performance of the subtyping module of the MALDI Biotyper system (Bruker Daltonik, GmbH) for the instant identification of KPC-producing Klebsiella pneumoniae, meticillin-resistant Staphylococcus aureus, and carbapenem-producing Bacteroides fragilis during routine workflow. We evaluated accuracy and impact on the turnaround time, with a particular focus on positive blood cultures. Further, we investigated the possibility to extend the subtyping for the detection of KPC specific marker to bacterial species other than K. pneumoniae.

Proteomics : Poster Presentations

Poster #1a in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Proteomics Red/Black Phosphorus and Phosphorene as Matrices and Calibrants in MALDI TOF Mass Spectrometry of Biomolecules Govinda Mandal - Masaryk University, Czech Republic *YI Grantee* ‣ Red and black phosphorus (P), as well as phosphorene, a monolayer or several layers of black phosphorus-prepared by ultrasound-added exfoliation in N-methyl-2-pyrrolidone3, or phosphorene-diamond composites, were examined as matrices in matrix-assisted laser desorption-ionization (MALDI) time-of-flight mass spectrometry of metabolites, peptides or other small molecules in complex mixtures. Performance of P-based matrices in LDI as well as in MALDI was examined, and the formation of Pn clusters suitable for internal mass calibration of complex biological samples was evaluated. Here, we demonstrate enhanced ionization of bio-molecules in complex biological liquids on unusual P matrices.

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Poster #1b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Proteomics Exploring the Effect of Urine Matrix Constituents on Recovery of Kidney Damage Markers in Urine Using a Targeted LC-MRM-MS Approach Tirsa van Duijl - Leiden University Medical Center *YI Grantee* ‣ Urine is a challenging and variable matrix for test development using a bottom-up proteomics strategy. We aimed to evaluate the effect of pre-analytical factors and matrix constituents on the quantification of urinary biomarkers TIMP-2 and IGFBP7 using LC-MRM-MS. Tryptic peptides from abundant non-target proteins induce ion suppression in ESI, resulting in reduced assay sensitivity of >400 % in samples with a protein concentration of >0.5 g/L. Non-specific proteolytic degradation of the peptide measurands by urinary proteases affected test recovery. Yet, non-specific degradation of urinary proteins could be prevented by thermal denaturation or by the addition of protease inhibitors. As the protease activity and total protein concentration is variable between patient urine samples, attention should be given to the pre-analysis in order to develop a robust and quantitative LC-MRM-MS test.

Poster #3d in Exhibit Hall - attended for 1 hr on Thursday starting at 12:30 Topic: Proteomics Lessons Learnt from the Fragmentation of Intact Glycopeptides Using Electron Transfer/Higher-Energy Collisional Dissociation (EThcD) Ádám Pap - Biological Research Center, Szeged *YI Grantee* ‣ A very complex mixture of intact, human glycopeptides was analyzed by LC-MS/MS. During MS/MS analysis a new fragmentation technique was used, electron transfer/higher-energy collisional dissociation (EThcD), which combines the advantages of ETD and HCD fragmentation. This work focuses on the fragmentation characteristics of glycopeptides during EThcD activation.

Poster #6c in Exhibit Hall - attended for 1 hr on Thursday starting at 10:00 Topic: Proteomics Targeted Proteomics Identifies Proteomic Signatures in Liquid-Biopsies of the Endometrium to Diagnose Endometrial Cancer Eva Coll de la Rubia - Vall Hebron Research Institute ‣ Endometrial cancer (EC) diagnosis relies on the observation of tumor cells in uterine aspirates, but it is associated with significant rates of undiagnosed patients and incorrectly diagnosed patients. We aimed to identify biomarker signatures in the fluid sample to overcome these limitations. The levels of 52 proteins were measured from two independent cohorts of patients of 38 and 116 patients (controls and EC including endometrioid and serous EC subtypes) by LC-PRM. Great values of sensitivity and specificity were achieved by a 2-panel signature for detecting EC cases and a 3-panel signature for the discrimination of EC subtypes. This study will improve diagnosis and assist in the prediction of the optimal surgical treatment.

Poster #8d in Exhibit Hall - attended for 1 hr on Thursday starting at 12:30 Topic: Proteomics Proteome Analysis of the Cervicaginal Fluid for Assessing the Severity of HPV-associated Intraepithelial Neoplasias Alexander Brzhozovskiy - V. I. Kulakov National Medical Research Center for *YI Grantee* ‣ The aim of this study is characterization of proteomic composition of the cervicaginal fluid for assessing the severity of HPV-associated intraepithelial neoplasias of the cervix among women of reproductive age. The study involved 80 volunteers who had various forms of HPV-associated cervical lesions (LSIL, HSIL and cervical cancer). The proteins associated with cervical lesions (LSIL, HSIL) and specific for cervical cancer were identified. A non-invasive approach based on cervicaginal fluid proteomic composition analysis for screening of HPV-associated intraepithelial neoplasias is developed. The method can be used for patient monitoring during treatment of various forms of HPV-associated cervical lesions.

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Poster #16a in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Proteomics Development of Parallel Reaction Monitoring (PRM) Assays for the Validation of Biomarkers Associated to Alzheimer’s Disease in CSF Claudia Fredolini - KTH – Royal Institute of Technology *YI Grantee* ‣ Suspension bead arrays (SBA) have shown to be a convenient and successful method for protein profiling of cerebrospinal fluid (CSF) in the contest of Alzheimer’s disease. Nevertheless, robust and specific orthogonal analytical methods are needed to support the verification of antibody based discoveries. We developed and validate parallel reaction monitoring (PRM) assays for 17 brain enriched proteins previously profiled by SBA in a cohort of 92 CSF samples from Alzheimer’s disease (AD) patients and control individuals. In order to verify the differential profiles observed by affinity proteomics, PRM assays were used to quantify the proteins in the same cohort.

Poster #16b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Proteomics A Comparative Study on Capillary and Venous Blood: Quantification of Proteins in Dried Blood Spots Eliška Stuchlíková - Masaryk University *YI Grantee* ‣ Dried blood spots (DBS) could be generated from capillary (cDBS) or venous (vDBS) blood and therefore the correlation of analyte levels in venous and capillary blood has to be evaluated. It was shown that levels of small molecules are different in cDBS vs. vDBS. However, limited data are available on protein concentration levels in cDBS and vDBS. In this study we have quantified and compared levels of three acute phase proteins alpha-1-antitrypsin, alpha-1-acid glycoprotein 1, alpha-1-acid glycoprotein 2, constitutive protein serum amyloid A4 and immunoglobulin A1 in cDBS and vDBS.

Poster #18b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Proteomics Mitochondrial Proteins as Parkinson’s Disease Circulatory Biomarkers – a Translational Study Sandra Anjo - University of Coimbra (CNC-UC) *YI Grantee* ‣ Using a translational approach (from cells secretome to plasma samples), we could identify two mitochondria-related proteins in plasma samples, which in combination lead to a powerful model with potential diagnostic value to discriminate the PD patients from matched. The analysis of secretomes from cells cultured under control or oxidative stress conditions, reveals several mitochondria-related proteins released in higher amounts under oxidative stress. This screening, performed by SWATH-MS, was translated to plasma samples from 28 control and 31 PD patients, and two of these proteins were found to be significantly changed. These proteins are associated with apoptotic mitochondrial changes, which may correspond to potential indicators of cell death and have never been reported as blood biomarkers for PD.

Poster #22g in Exhibit Hall - attended for 1 hr on Thursday starting at 10:00 Topic: Proteomics Plasma Proteome Profiling to Detect and Avoid Sample-related Biases in Biomarker Studies Philipp Geyer - Department of Proteomics and Signal Transduction, ‣ Recently, we have developed an automated and robust shotgun proteomics pipeline – called `Plasma Proteome Profiling` – to yield a proteomic reflection of an individual´s phenotype. So far, we have applied this workflow to different health and disease studies. Here we ask, if we can use the pipeline to determine the quality of plasma samples. We carried out experiments resulting in three different quality marker panels that allow assessment on the level of the individual samples and of entire studies, both on a global scale. Furthermore, our recently developed global correlation analysis allowed us to determine if biomarker candidates belong to one of the three quality marker panels. The relevance of the quality marker panels is demonstrated in a literature analysis of more than 200 publications, half of which reported quality markers as biomarker candidates.

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Poster #24e in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Proteomics Study of the Human Tear Proteome as a Possible Biomarker Source Bella Bruszel - University of Szeged ‣ Human tear is a relatively easily collectible body fluid which contains several types of compounds what makes it a possible biomarker source. The human tear however has several functions and according to it its composition and volume show large dynamic variations. Because of this property of tear fluid a question emerged: could be a single sample from a randomly chosen single eye at any day a representative sample for the tear proteome of a healthy person? In our work using a shotgun label free LC-MS method we wanted to find the answer to this question by examining the qualitative and quantitative intra- and interpersonal variances of tear proteome. High intra- and interpersonal variances were found in the total protein content and in the relative amount of the different proteins. Because of this observation more samples are needed for a correct statistical evaluation.

Small Molecules : Poster Presentations

Poster #1d in Exhibit Hall - attended for 1 hr on Thursday starting at 12:30 Topic: Small Molecules Easy and Fast Analysis of Vitamin K1 in Serum Using 2-Dimensional-UPLC-MS/MS Ida Bøgh Andersen - Uni.of Southern Denmark,Lillebaelt Hospital, Vejle *YI Grantee* ‣ Vitamin K1 is a sub-group of the fat-soluble very non-polar vitamin Ks. The non-polar nature has given rise to several challenges during method development, because vitamin K1 sticks to materials used during the process and is lost during evaporation. We found that reducing the sample preparation as much as possible offline, instead using 2-dimensional UPLC-MS/MS improves recovery and gives satisfactory chromatograms. Moreover, the method is simple and fast. We have now validated the method for vitamin K1, and validation details will be presented.

Poster #2b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Small Molecules Quantification of Antimycotics in Human Plasma or Serum by LC-MS/MS for Clinical Research Claudio De Nardi - Thermo Fisher Scientific ‣ A clinical research analytical method for the quantification of 8 antimycotics in human plasma or serum was implemented on a Thermo Scientific™ Vanquish™ Flex Binary LC system connected to a Thermo Scientific™ TSQ Quantis™ triple quadrupole mass spectrometer. Detection was performed by selected reaction monitoring (SRM) using an isotopically labeled internal standard for each target analyte. Method performance was evaluated using the ClinMass® TDM Platform with the ClinMass Add-On Set for Antimycotics from RECIPE Chemicals + Instruments GmbH (Munich, Germany) to obtain linearity of response within the calibration ranges, accuracy, and intra- and inter-assay precision for each analyte.

Poster #2c in Exhibit Hall - attended for 1 hr on Thursday starting at 10:00 Topic: Small Molecules Towards the Discovery of Markers in Parkinson’s Disease: Investigation of Trace Amines and Related Metabolites Antonina Gucciardi - University of Padova *YI Grantee* ‣ This pilot study evaluated whether circulating profiles of catecholamines, trace amines and indoles were altered in early and late stages of Parkinson’s disease. Plasma samples were collected from 48 PD patients, of which 21 untreated de novo and 27 treated patients, and 10 healthy subjects and metabolites were quantified by UPLC-MS/MS. Significantly different circulating levels of tyrosine, tyramine, synephrine, norepinephrine, metanephrine, β-phenylethylamine and serotonin were found. Our findings suggest that PD is characterized by profound changes in the aminergic and indolic neurotransmitters. Compounds within these families may constitute putative markers for early stage detection and progression of PD.

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Poster #4a in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Small Molecules Analysis of Thiamine Pyrophosphate and Pyridoxal-5’-Phosphate in Whole Blood Fully Automated Sample Preparation LC-MS/MS System (CLAM-2000 + LCMS-8045) Dennis van den Heuvel - Shimadzu Benelux ‣ Thiaminepyrophosphate and Pyridoxal-5’-phosphate in whole blood are predominantly analysed with HPLC and fluorescence detection with excessive sample preparation including derivatisation. The aim of this study was to set up an automated solution for the analysis of TPP and PLP in whole blood samples, using the vitamin B1 & B6 LC-MS/MS kit from Instruchemie in combination with the CLAM-2000 coupled to the Nexera X2 UHPLC LCMS-8045 (Shimadzu Corporation). Both compounds showed good linearity (r2> 0.995) in a clinically relevant concentration range and acceptable within and between precision and accuracy.

Poster #4c in Exhibit Hall - attended for 1 hr on Thursday starting at 10:00 Topic: Small Molecules Vitamin D in Veterinary Health and Disease: Development of an LC-MS Assay for Clinical and Research Use Emma Hurst - University of Edinburgh *YI Grantee* ‣ Vitamin D status is linked to several non-skeletal health disorders in veterinary species. The current challenge is to define whether vitamin D is causally linked to the initiation, development and outcome of these conditions, or whether it is simply a marker of ill-health. Here, I will discuss the development a LC-MS assay to accurately quantify several vitamin D analytes in a variety of species. This will allow us to answer key research questions about the role of vitamin D analytes in disease development, and to potentially impact veterinary clinical care by investigating the potential role of vitamin D in the treatment of important diseases.

Poster #4d in Exhibit Hall - attended for 1 hr on Thursday starting at 12:30 Topic: Small Molecules Clam- SPE on Line- UHPLC-MS/MS : Fully Automatic, Simultaneous and Quickly Quantification of Drugs of Abuse in Blood and Saliva Maria Rosaria Innarella - Shimadzu Europe ‣ The developed method is able to automatically quantify various drugs of abuse in blood and saliva matrix on Floqswab®, through by an online sample preparation and cleaning with Clam-2000 and SPE. The calibration curve is determined between 0.5 ng/mL and 500 ng/mL. The achieved results in both matrix have a good accuracy between 85 and 115%. The ion ratio stays stable with relative response which doesn’t exceed 30%. The repeatability is determined by spiking 5 samples in blood and 5 saliva on Floqswab® to the limit of quantification They are estimated to be less than 15%.

Poster #5a in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Small Molecules UPLC-MS/MS Method For Rapid Urinary GHB Confirmation Leen Mortier - Jessa Hospital, Clinical Laboratory ‣ Confronted with questionable results from a dipstick test for the DoA gamma-hydroxybutyric acid (GHB) in urine and possible major patient implications of false results, an UPLC-MS/MS confirmatory method based on direct isotope dilution analysis (DIDA) was validated and implemented. A dilute-and-shoot approach was combined with chromatographic separation by gradient elution in the reversed phase mode and MS detection in the ES negative mode (3 MRM transitions). Imprecision and accuracy were acceptable at relevant concentrations, LOD was 2 mg/L. Alpha- and beta-hydroxybutyric acid were chromatographically separated. Matrix effects were substantial but were compensated by the deuterated internal standard. We thus implemented a fit-for-purpose UPLC-MS/MS method for confirmation of urinary GHB levels pointing to exogenous ingestion/administration (cutoff 10 µg/mL).

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Poster #5b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Small Molecules Analysis of DHT, DHEA, Testosterone, Androstenedione, 17-OHP and Progesterone by LC-MS/MS for Clinical Research Robert Wardle - Waters Corporation ‣ An offline automated method for the measurement of serum dihydrotestesterone (DHT), dehydroepiandrosterone (DHEA), testosterone, androstenedione, 17-hydroxyprogesterone (17-OHP) and progesterone has been evaluated, enabling steroid profiling for the investigation of metabolic dysfunction biomarkers for clinical research. An LC-MS/MS method was developed using a mixed-mode Solid Phase Extraction (SPE) sorbent in 96-well plate format, improving workflow and reducing sample preparation time. Chromatographic resolution between structurally related steroid species was achieved using a reversed phase C18 UPLC column. This offline automated method demonstrates excellent linearity, analytical sensitivity, selectivity, precision and accuracy, while providing high sample throughput capabilities. For Research Use Only, Not for use in diagnostic procedures.

Poster #6d in Exhibit Hall - attended for 1 hr on Thursday starting at 12:30 Topic: Small Molecules LC-MS/MS Method for the Quantification of Nine Antiepileptic Drugs from Dried Plasma Spots and Their Long Term Stability Ugo de Grazia - Fondazione IRCCS Istituto Nazionale Neurologico Ca ‣ A method for the determination of nine commonly prescribed AEDs in plasma spotted on DSSD was developed and long term stability evaluation was carried out. 50 µl of plasma were distributed on DSSD then the device was dried and stored at different temperatures. Drugs were extracted from the device, separation was performed on a UHPLC C-18 RP column and quantified using a triple quadrupole mass spectrometer. The method was validated considering the concentration ranges encountered in the routine clinical practice. The assay was linear over the concentration ranges tested. Recovery ranged from 93.7% to 106.8%. Precision ranged from 2.1% to 18.4% while accuracies ranged from 88.4% to 113.9%. No matrix effects was found. At temperatures tested stability results showed that the mean differences were comprised between -3.4 and -12.9% after a 15-day storage period on DSSD.

Poster #8c in Exhibit Hall - attended for 1 hr on Thursday starting at 10:00 Topic: Small Molecules An UHPLC-MS/MS Method with Protein Precipitation to Quantify Fourteen Antihypertensive Drugs and Two Metabolites in Human Plasma: Update for Adherence Testing Amedeo De Nicolò - University of Turin ‣ One of the main problems in the management of resistant hypertension (RH) is the discrimination of real cases of RH from poor therapeutic adherence. In this work our group validated a method for the quantification of fourteen antihypertensive drugs and two metabolites in human plasma within the same run with high sensitivity performance. Considering the high usefulness and reliability of TDM for adherence testing, this method is expected to be widely applied in the near future.

Poster #9c in Exhibit Hall - attended for 1 hr on Thursday starting at 10:00 Topic: Small Molecules Fully Automated LC-MS/MS Analysis of Anticoagulants Using a Novel Reagent Kit Sigrid Baumgarten - Alsachim ‣ The screening and precise quantitation of several Anticoagulants for therapeutic purpose is clinically relevant to avoid any hemoragy. In order to develop a multi-analyte approach, we have used a novel reagent kit. To allow an easy screening by tandem MS for the panel of anticoagulants targeted, an automated sample preparation robot directly coupled to the LCMS system was used. By simply placing plasma collection tubes in the system, the analysis is performed through the LCMS analysis automatically.

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Poster #12a in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Small Molecules Fully Automated Analysis of Immunosuppressant Drugs in Whole Blood using Stable Labeled Isotope Internal Standards Fanny Dayot - Alsachim ‣ The Quantitation of Immunosuppressant for therapeutic purpose is clinically relevant after organ transplantation. In order to develop a multi-analyte approach, we have used a novel reagent kit. The analysis of Immunosuppressant drugs was performed by simply placing whole blood collection tubes in the fully automatic preparation unit online with a LCMS system. The fully automatic LCMS preparation unit was programmed to perform sample extraction and protein precipitation followed by filtration and sample collection. Then the extracted samples were analyzed in 1.4 min.

Poster #12b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Small Molecules Comparison of Sample Preparation Options for the Extraction of a Panel of Endogenous Steroids from Serum Prior to UHPLC-MS/MS Analysis Alan Edgington - Biotage GB Limited ‣ This poster compares sample preparation options for the extraction of a panel of endogenous steroids from serum. Method parameters were optimized for increased sensitivity: MRM transitions, chromatography and mobile phase additives due to the necessity of positive and negative ionisation modes. LC-MS/MS analysis was performed using a Shimadzu Nexera UHPLC system coupled to an 8060 triple quadrupole MS. Solid phase extraction was compared to supported liquid extraction and methods optimized for a steroid panel including and without DHEAS. Final optimized methods were compared for extract cleanliness by investigation for phospholipid content.

Poster #13a in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Small Molecules Plasma Free Metanephrines Measured by LC-MS/MS: A New Diagnostic Tool for the Diagnosis of Neuroblastoma Giuliana Cangemi - PhD, Istituto Giannina Gaslini ‣ In this work we have investigated the diagnostic role of plasma free metanephrines (PFM):metanephrine (MN), normetanephrine (NMN) and 3-methoxytiramine (3-MT)quantified by LC-MS/MS, for neuroblastoma (NB), the most common extra-cranial solid tumor in children. 3-MT and NMN showed an excellent diagnostic performance. The determination of PFM should be taken in consideration as a new diagnostic and prognostic tool and further validated in clinical prospective studies in comparison to urinary catecholamines and metanephrines.

Poster #14d in Exhibit Hall - attended for 1 hr on Thursday starting at 12:30 Topic: Small Molecules Cascadion™ SM Clinical Analyzer: The First Fully Integrated LC-MS/MS Analyzer for the Clinical Laboratory Torsten Binscheck-Domaß - Thermo Fisher Scientific Clinical Mass Spectrometry ‣ Coupling high performance liquid chromatography to quadrupole based mass spectrometry (LC-MS/MS) has been proven to be a valuable technology to address analytical challenges in clinical laboratories. However the limited throughput of samples, the need for highly qualified operators and the low level of automatization prevents clinical LC-MS/MS from entering laboratory routine. As a first and IvD certified instrument the Cascadion™ SM Clinical Analyzer is designed to close the gap between demands for analytical performance and the needs of clinical laboratories. It is discussed, how the Cascadion™ SM Clinical Analyzer can improve the quality of patient care.P roduct not 510(k) cleared and not yet available for sale in the U.S.

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Poster #15d in Exhibit Hall - attended for 1 hr on Thursday starting at 12:30 Topic: Small Molecules GC-NCD-QTOF Dual Detector in Quantitative Screening of 40 Stimulant-Type New Psychoactive Substances in Urine and Blood without Using Reference Standards Samuel Mesihää - University of Helsinki, Department of Forensic Med *YI Grantee* ‣ Analysis of the new psychoactive substances (NPS) is a challenge to the clinical toxicology laboratories because it is difficult to obtain an authentic reference standard in short period of time, while these compounds are still being consumed. An analytical platform was developed for the simultaneous identification and quantification of NPS by using a GC system that directs the gas flow to quadrupole time-of-flight mass analyzer (QTOF) and nitrogen chemiluminescence detector (NCD). In this system, the drug screening is based on the detection of the accurate masses of the protonated molecular ion (MH+) produced in the atmospheric pressure chemical ionization source. Quantification is based on the equimolar response of drugs to nitrogen. In this work, GC-NCD-QTOF detector used to measure 40 stimulant-type NPS spiked in the blood and urine samples.

Poster #16d in Exhibit Hall - attended for 1 hr on Thursday starting at 12:30 Topic: Small Molecules Drugs Of Abuse 24/7 Monitoring Frank Streit - UMGL UMG Goettingen ‣ For clinical toxicological screening polyclonal antibodies are most common in emergency laboratories. Diverse cross reactivity results in a large number of false positive or false negative determinations. Up till now the 24/7 emergency analysis of these drugs is restricted to a certain number of special skilled staff with chromatographic and mass spectrometry expertise and therefore not available. Personal costs for implementing such an emergency service are high. The aim of this study was to establish a standardized methodology for screening up to 150 common used drugs with a fully automated LC-MS/MS system equipped with an integrated sample preparation module (CLAM-2000, Shimadzu). We validated antidepressive drugs, AEDs, benzodiazepines as well as neuroleptic drugs in one quantitative 6min screening method.

Poster #16e in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Small Molecules Isotope Dilution Turboflow™ LC-HRMS for the Simultaneous Quantification of 12 Antimycotics in Human Serum Carina Schuster - University Hospital LMU Munich ‣ The aim of this study was to develop and validate an isotope-dilution-TurboFlow™-high resolution mass spectrometry (MS) method for therapeutic drug monitoring of the 12 most widely used systemic antimycotics. With a short turn-around time the proposed method provides a simple and robust sample clean-up minimizing the interference of the sample matrix. Application of commercially available kit components allows for handling high level of convenience.

Poster #17a in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Small Molecules Fully Automated Solution for Therapeutic Drug Monitoring of over 150 Drugs in Serum or Plasma with LC-MS/MS Analysis Katharina Kern - RECIPE Chemicals + Instruments GmbH ‣ We report a fully automated solution for the RECIPE ClinMass® TDM Kit System for LC-MS/MS analysis of over 150 drugs and metabolites in serum or plasma by the use of a Tecan® Freedom EVO® liquid handling system. The sample preparation procedure was automated by the use of 96-well filter plates. Data was acquired with a Thermo ScientificTM EnduraTM and an Agilent 6460 triple quadrupole mass spectrometer. Precision and accuracy testing showed outstanding results for all analytes. Values obtained with the automated approach also showed excellent correlation with manual sample preparation results.

Poster #17e in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Small Molecules Diagnosis of Gout and Pseudogout Using Inorganic TiO2 Matrices for LDI-TOF Mass Spectrometry Moon-Ju Kim - Yonsei University *YI Grantee* ‣ Gout and pseudogout are two types of crystal-induced arthropathies. Because they share similar symptoms, it"s very difficult to differentiate these two diseases from the observation of signs. Therefore, accurate diagnosis is critical for the appropriate treatment. Although finding crystals in synovial fluid has been the gold standard for the diagnosis of these disorders, it contains several problems such as time-consuming analysis and high false negative results. In this work, we introduced a new diagnostic approach using LDI-mass spectrometry with inorganic nanostructures as solid matrices. It includes the quantification of arthropathy-triggering crystals as well as the differentiation of two similar diseases.

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Poster #18a in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Small Molecules Analysis of Antiepileptic Drugs in Human Serum Using an Ultivo LC/TQ Carrie Adler - Agilent Technologies ‣ Analyzing antiepileptic drugs can be challenging in clinical research due to the disparate concentrations at which these drugs may be present in human serum. Therefore, a quality assay must be able to analyze many compounds simultaneously, over several orders of magnitude. A highly sensitive and specific analytical method for the quantitation of 15 antiepileptic drugs in human serum was tested on an innovative miniature triple quadrupole mass spectrometer, the Agilent Ultivo triple quadrupole LC/MS (LC/TQ). Samples were prepared through a simple protein precipitation/dilution protocol. Analytes could be quantified over a wide dynamic range; accuracy and reproducibility metrics, as well as R2 values, were acceptable. For Research Use Only. Not for use in diagnostic procedures.

Poster #18c in Exhibit Hall - attended for 1 hr on Thursday starting at 10:00 Topic: Small Molecules Integration of Mycophenolate and its Metabolite Analysis in Plasma Using LC-MS/MS with Full-Automated Sample Preparation Stephane Moreau - Shimadzu Europa GmbH ‣ The main goal of this research was to develop a simple, rapid and sensitive LC-MS/MS method with automated sample preparation unit for simultaneous quantification of mycophenolic acid and its glucuronide in human plasma. Sample extraction was carried out using a simple protein precipitation automatically. The extracted samples were analyzed with a trap and elute method in 2 min. Method linearity was established in the concentration range of 0.1 to 50 mg/L for mycophenolic acid and 1 to 250 mg/L for its glucuronide. Repeatability, reproducibility and accuracy of these compounds were well within the acceptance criteria specified in the guidelines such as ICH and EMA.

Poster #18d in Exhibit Hall - attended for 1 hr on Thursday starting at 12:30 Topic: Small Molecules Quantification of Antiepileptics in Human Plasma or Serum by LC-MS/MS for Clinical Research Magnus Olin - Thermo Fisher Scientific ‣ A clinical research analytical method for the quantification of 25 Antiepileptics in human plasma or serum was implemented on a Thermo Scientific™ Vanquish™ Flex Binary LC system connected to a Thermo Scientific™ TSQ Quantis™ triple quadrupole mass spectrometer. Detection was performed by selected reaction monitoring (SRM) using 20 deuterated internal standards. Method performance was evaluated using the ClinMass® TDM Platform with the ClinMass Add-On Set for Neuroleptics from RECIPE Chemicals + Instruments GmbH (Munich, Germany) to obtain limits of quantification, linearity ranges, accuracy, and intra- and inter-assay precision for each analyte.

Poster #19a in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Small Molecules Analysis of Phosphatidylethanol in Human Whole Blood by LC-MS/MS Frances Carroll - Restek Corporation ‣ Phosphatidylethanol is a group of phospholipids formed through enzymatic reaction between ethanol and phosphatidylcholine on the cell membrane. Among multiple homologues of PEth, PEth-16:0/18:1 (palmitic acid/oleic acid) is the predominant molecule extracted from human erythrocytes and can be measured in whole blood as specific biomarker of alcohol consumption with a detection window of up to 3-4 weeks and with the application of highly sensitive LC-MS/MS techniques, it is now possible to use PEth concentration in blood to differentiate chronic drinking from social drinking or as a marker of absolute abstinence. In this study, a fast chromatographic analysis was developed using a Raptor FluoroPhenyl column. Specific and sensitive measurement of PEth-16:0/18:1 in whole blood was achieved with a combination of simple protein precipitation and fast 3.5-minute LC cycle time.

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Poster #19d in Exhibit Hall - attended for 1 hr on Thursday starting at 12:30 Topic: Small Molecules A Novel LC-MS/MS Quantification Method for Amino Acids in Human Plasma, including Alloisoleucine, without Ion Pairing or Derivatization Aurore Jaffuel - Shimadzu France ‣ Amino acids are routinely assayed to diagnose inherited metabolism disorders. Recently, a new LCMSMS method was developed, without derivatization or ion paring, for the high sensitive quantification of amino acids (A. Jaffuel, poster, MSACL EU 2016, Salzburg). However a separate injection was needed for alloisoleucine quantification. Alloisoleucine is of much interest as currently the most specific marker for MSUD. We here present a new method for the quantification of 50 amino acids, including the separation of leucine, isoleucine and alloisoleucine. Total cost per sample is reduced from 13€ to 2€ compare to current methods. Plasma controls showed good accuracies.

Poster #20a in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Small Molecules High-Throughput Simultaneous Measurement of Vitamins A, D and E in Human Plasma by LDTD-MS/MS. Ichiro HIRANO - SHIMADZU Corporation, MS Business Unit ‣ Epidemiologic studies to asses nutritional status of population, require high-throughput and multiplexed analysis of small sample volumes.fat-soluble vitamins have several roles to maintain good health and normal development. In order to face the large number of samples, a LDTD/MS/MS method has been developed to assay vitamins A, D and E in human serum. The method was partially validated and compared to a LC/MS/MS to check if the lack of separation was affecting the results.

Poster #21b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Small Molecules Antiepileptic Drugs 24/7 Monitoring Gry Helene Dihazi - Clinical Chemistry, University Medicine Goettingen ‣ Epilepsy is one of the most common neurological disorder affecting people of all ages. Various factors may influence plasma concentrations of antiepileptic drugs (AEDs). TDM of AEDs may be useful for individualizing the therapy to aid the treatment of uncontrollable seizures and clinical toxicity. The aim of this study was to develop and to implement a methodology for 26 different AEDs with a fully automated LC-MS/MS system equipped with an integrated sample preparation module (CLAM-2000, Shimadzu) and to test the feasibility of a 24/7 monitoring service by technicians without prior chromatographic or mass spectrometric experience. Robustness, excellent precision and accuracy, which were confirmed in our evaluation study, were convincing. In addition, the user-friendly control panel and familiar user interface of the CLAM-2000 provide 24-hour LC-MS for emergency analyses.

Poster #22a in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Small Molecules Determination of Eicosanoids by UHPLC-MS/MS in Diluted Interstitial Fluid Obtained with Open Flow Microperfusion (OFM) Anita Eberl - Joanneum Research ‣ This study describes the quantification of eicosanoids in dermal interstitial fluid (dISF) samples using UHPLC-MS/MS. The dISF samples were obtained by using dermal open flow microperfusion (dOFM) in a rat model of skin inflammation. Eicosanoids were significantly increased in inflamed skin sites that were treated with imiquimod relative to untreated control sites. In addition, oral treatment with an anti-inflammatory glucocorticoid decreased eicosanoid concentrations. Results show that a combination of tissue-specific sampling with LC-MS analytics is well suited to analyze small sample volumes from minimally invasive sampling methods to study local inflammation and the effect of treatments in skin diseases.

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Poster #22b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Small Molecules A Novel Solution for Vitamin K1 and K2 Analysis in Human Plasma by LC-MS/MS Hansjoerg Majer - Restek Corporation ‣ Vitamin K is a group of fat-soluble vitamins divided into vitamin K1 (one compound, phylloquinone) and K2 (a group of compounds, menaquinones). As an interest for their biological action in extra-hepatic tissues is increasing, an accurate and simple measurement of vitamin K status remains a critical issue for both clinical research and diagnostics. There is a high degree of analytical challenges in vitamin K analysis, as it is the most lipophilic and least abundant of the fat-soluble vitamins. In this study, a simple and fast plasma sample preparation procedure was developed using phospholipid removal in combination with a chromatographic analysis using a Raptor Biphenyl column. The method provides a novel solution towards vitamin K research and high-throughput clinical diagnosis.

Poster #22f in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Small Molecules A UPLC-MS/MS Method for the Quantification of Serum Methylmalonic Acid (MMA) Craig Livie - Glasgow Royal Infirmary *YI Grantee* ‣ Deficiency in vitamin B12 can lead to severe symptoms in patients, including neuropsychiatric changes, therefore timely and accurate detection is vital. Detection of total B12 from serum is an unreliable measure of B12 status in patients. Methods which accurately measure alternative markers of B12 such as methylmalonyl CoA (MMA) are therefore urgently needed for use in clinical laboratories. We established an LC-MS/MS method for measurement of serum MMA developed in collaboration with Waters. The method described demonstrated robust performance on reverse phase chromatography with SPE in-well protein crash.

Poster #23b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Small Molecules Introducing a LC-MS/MS Method for the Screening of Anti-Hypertensive Drugs in Urine Melissa McNaughton - University of Manchester *YI Grantee* ‣ A liquid chromatography-tandem mass spectrometry method to measure anti hypertensive drugs in urine was set up using the Waters Acquity TQD. The method was set up to allow measurement of a total of 29 drugs which are currently prescribed to NHS Lothian patients for hypertension.

Poster #24c in Exhibit Hall - attended for 1 hr on Thursday starting at 10:00 Topic: Small Molecules Volumetric Absorptive Microsampling as an Alternative Sampling Strategy for Cerebrospinal Fluid Lisa Delahaye - Ghent University *YI Grantee* ‣ Dried matrix spot (DMS) sampling is an alternative sampling strategy with typical sample volumes less than 50µL, with many benefits over traditional sampling. For cerebrospinal fluid (CSF), the number of applications making use of DMS is limited. Using a microsampling strategy like volumetric absorptive microsampling (VAMS) allows to collect larger numbers of samples from patients. To date, VAMS has not yet been used for the analysis of CSF. As a proof of concept for CSF microsampling via VAMS, we chose paracetamol as model compound. Methods were developed and validated for the quantification of paracetamol in dried blood and dried CSF utilizing VAMS. This method is the first to quantify analytes in dried CSF samples obtained via VAMS. As the knowledge of paracetamol levels in CSF and its correlation with blood levels is limited, this method could be used for pharmacokinetic research.

Poster #24d in Exhibit Hall - attended for 1 hr on Thursday starting at 12:30 Topic: Small Molecules Quantification of Δ9-Tetrahydrocannabinol and Cannabidiol in Plasma and Decoctions by LC-MS/MS: Application for TDM of Medical Cannabis Sebastiano Barco - Istituto Giannina Gaslini *YI Grantee* ‣ Medical cannabis has being increasingly administered as medical therapy to treat disease or alleviate symptoms conditions. Monitoring of blood levels of Δ9-tetrahydrocannabinol and cannabidiol is necessary for assessing pharmacokinetic parameters in order to optimize drug administration. Due to low concentration in plasma and possible interference by other cannabinoid present both in biological sample and in the galenic preparations, we propose a new UHPLC-MS/MS method of analysis.

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Tissue Imaging : Poster Presentations

Poster #11b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Tissue Imaging Biomedical Applications of the Ambient Mass Spectrometry: Myths and Realities Vladimir Frankevich - National Research Center for Obstetrics and Gynec ‣ A direct analysis of the physiological processes in living organisms remains a desirable but so far, an unreachable goal for mass spectrometry (MS). The conventional MS implies the destructive sample preparation routines and as such has limited applicability for the in-vivo experiments. Yet, with a development of the ambient ionization techniques the door to a direct in-vivo analysis could be opened. In this presentation we give recent results of new techniques and their applicability to the in vivo analysis of the biological material such as e.g. malignant tissues.

Poster #15a in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Tissue Imaging Structural Polymorphism in Evolving Amyloid Plaque Pathology is Associated with Distinct Amyloid-Beta (Aβ) Truncation Profiles Wojciech Michno - Sahlgrenska Academy, University of Gothenburg ‣ Alzheimer's disease (AD) is characterized by accumulation of amyloid-β (Aβ) peptides into different extracellular plaques. Plaques have also been found in non-demented pathological ageing patients. Therefore, discrimination between structural and molecular plaque architecture are of essential interest to resolve Aβ plaque pathology in AD. Here, electrooptic fluorescent probes that recognize structural Aβ polymorphism, laser microdissection, and MALDI IMS were utilized to probe Aβ peptides in morphologically distinct Aβ plaques, in AD, cognitively normal PA, and transgenic AD mice. Human based study revealed cored deposits to be associated with presence of Aβ1-40, while no Aβ1-40 was observed in diffuse plaques in AD or PA. The AD mice data revealed further that the increase in Aβ1-40 was associated with plaque maturation over time.

Poster #15e in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Tissue Imaging Three Dimensional MALDI-Imaging Mass Spectrometry Mapping of a Calcified Mouse Heart Stephanie Mezger - M4I, Maastricht University *YI Grantee* ‣ Cardiac ischemia may cause myocardial injury followed by cardiac remodelling and dystrophic calcification, this process is not completely understood yet. With this project we aim to establish a (3D) spatial map of lipids, metabolites and peptides using matrix assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) on a calcified mouse heart to understand the physiological mechanisms that leads to the process of calcification. Our data reveals specific molecular profiles in the calcified and non-calcified regions, suggesting a switch in the metabolic pathways that occur with remodelling of the heart in the process of calcification. Complementary protein and peptide analysis will help to understand the underlying biological mechanisms related to myocardial injury.

Poster #15g in Exhibit Hall - attended for 1 hr on Thursday starting at 10:00 Topic: Tissue Imaging Cross-Modality Correlation of Multimodal Imaging Mass Spectrometry Data at Single-Pixel Resolution Patrick Wehrli - University of Gothenburg *YI Grantee* ‣ Our group has previously been investigating neuropathology associated distribution patterns of lipids and proteins in transgenic mouse models of Alzheimer’s disease (AD) using multimodal imaging strategies.1 In here, were present a continuation of these efforts by means of a data processing routine that enables direct correlation of chemical information across multiple imaging mass spectrometry (IMS) modalities. Here brain sections of transgenic AD mice (tgSwe) were analyzed using matrix-assisted laser desorption/ionization (MALDI) IMS in three modalities: lipids in (1) negative and (2) positive ion mode, and (3) peptides in positive ion mode. The three IMS data cubes were processed by spectral pretreatment and co-registration prior to multivariate image data analysis. This strategy enabled direct and unsupervised cross-modality correlation at the measured image resolution.

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Poster #17f in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Tissue Imaging Evaluation of Different Embedding Methods for Fragile and Small Size Clinical Samples in MSI Analysis Emine Kazanc - Imperial College of London *YI Grantee* ‣ Mass spectrometry imaging (MSI) is a powerful technique which able to provide detail information on the spatial distribution of metabolites, drug molecules and lipids in tissue sections. However, sectioning of fragile and small specimens can be challenging for MSI analysis. Embedding multiple small specimens allow simultaneous sectioning but requires an optimum embedding medium. This study aims to compare different polymers and freezing methods to develop a comprehensive tissue embedding protocol for the best MSI analysis.

Poster #24a in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Tissue Imaging Dual Polarity Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging (MALDI-MSI) of Focal Cerebral Ischemic Rat Brain Using 1,5-Diaminonaphthalen Sohee Yoon - Korea Research Institute of Standards and Science ‣ A matrix application was constructed to perform matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) for lipid analysis of positive ions, negative ions, and MS/MS in a single tissue. Focal cerebral ischemia-induced rat brain tissue was imaged to profile lipids associated with the disease. The specific lipids profiled included ceramide, phosphatidylcholine, lysophosphatidylcholine, and N-acyl-phosphatidylethanolamine. In particular, it is the first MALDI-MSI report has demonstrated that NAPEs are lipids related to focal cerebral ischemia. Here, we discussed the analytical method that can be perform dual polarity and MS/MS in the same tissue under same condition by using a single matrix.

Troubleshooting : Poster Presentations

Poster #25a in Exhibit Hall - attended for 15 min on Tuesday starting at 19:00 Topic: Troubleshooting What Happened to My Topiramate? Magdalena Rajska - SPADIA Lab, a.s. ‣ We ran into a problem with spiked calibrations due to technical problem with pipets. Until technical problem can be solved a commercial calibrator set for Antiepileptics in serum assay was purchased. When evaluated with commercial calibration 6 of 7 antiepileptics met IQC requirements, topiramate did not. We checked for potential changes/errors in acquisition and evaluation methods, when those factors were ruled out we thought there may have been an error in ClinCal®-Calibrator list but further troubleshooting steps reveled that chosen ESI+ MRMs for TOPI and TOPI-d12 are probably matrix and gradient effected. When ESI- MRMs for TOPI were used, topiramate met IQC requirements for both, spiked and commercial calibration.

Poster #25b in Exhibit Hall - attended for 15 min on Tuesday starting at 19:15 Topic: Troubleshooting Optimizing Chromatographic Conditions for HILIC Analysis of Catecholamines Gareth Hammond - Waters Corporation ‣ Using HILIC chromatography for very polar analytes is prone to peak tailing, due to low ionic strength mobile phases and immiscibility of organic mobile phases with high salt concentration aqueous mobile phases. Here we describe the use of mobile phase preparation protocols, which ensure full mobile phase miscibility and high salt concentration solubility, when using HILIC chromatography for the separation of catecholamines. For Research Use Only. Not for use in diagnostic procedures.

Poster #25c in Exhibit Hall - attended for 15 min on Tuesday starting at 19:30 Topic: Troubleshooting An Alternative Solution to High Blank Problems Anne Schmedes - Department of Clinical Biochemistry ‣ When setting up new analysis for drugs or endogenous compounds a gradient separation on a UPLC column is often used before tandem mass spectrometry. However on two occasions we have experienced that it can be impossible to have clean blank samples when running a gradient. Two examples will be shown: 1) analysis of Caffeine and 2) analysis of Erlotinib. For both these compounds we experienced that mobile phases inevitably contain the compounds or similar compounds. The only way to obtain acceptable blank samples was to use isocratic elution.

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Poster #26a in Exhibit Hall - attended for 15 min on Wednesday starting at 12:30 Topic: Troubleshooting Metformin Interference in LC-MS/MS Analysis of Plasma Methoxycatecholamines Marianne Bergmann - Lillebaelt Hospital, Vejle ‣ The LC-MS/MS analysis for plasma methoxycatecholamines has been in routine use at Lillebaelt Hospital since October 2015. During the spring of 2017 we experienced an increasing problem with sample chromatograms showing lower peak height for metanephrine and d3-metanephrine, but not for normetanephrine and d3-normetanephrine. The problem only affects a few samples in each run, but these samples also show poor peak shape. We discovered that the problem was caused by metformin – a drug used to treat type 2 diabetes. Metformin co-elutes with metanephrine and causes major ion suppression. Because metformin is widely used and in high doses (up to 2000 mg/day), we set out to solve this problem.

Poster #26b in Exhibit Hall - attended for 15 min on Wednesday starting at 12:45 Topic: Troubleshooting Charge Wars of Ion Suppression - Awakening the Force for the Analysis of Estrogens in Clinical Research Robert Wardle - Waters Corporation ‣ Ion suppression in LC-MS/MS methods can be considered the dark side of the force, possibly leading to analytical sensitivity, selectivity and accuracy goals not being met. Here we discuss the routine measurement of 17β-Estradiol (E2) and Estrone (E1) for clinical research as an example. Testing was performed to investigate the source of a disturbance in the force (labware, biological matrix or solvents), chromatographically resolve it from the analytes and identify the interferent using MS scanning techniques and accurate mass analysis. For Research Use Only, Not for use in diagnostic procedures.

Poster #26c in Exhibit Hall - attended for 15 min on Wednesday starting at 13:00 Topic: Troubleshooting Cortisol Everywhere! Neus Fabregat-Cabello - University of Liège, CHU de Liège *YI Grantee* ‣ Our problem with a routine LC-MS/MS system is related to the presence of an isobaric interference of cortisol from an unknown source. After cleaning and checking the proper function of the LC system, we performed an exhaustive study of all the possible contamination sources that can affect the method performance. Bovine Serum Albumin (BSA) seems the critical reactive that contained the highest amount of this interference, among other sources tested. Identity of this compound was obtained by QTOF analysis and resulted to be cortisol.

Various Other : Poster Presentations

Poster #2a in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Various Other Quantification of Neuroleptics in Human Plasma or Serum by LC-MS/MS for Clinical Research Madalina Oppermann - Thermo Fisher Scientific ‣ A clinical research analytical method for the quantification of 28 neuroleptics in human plasma or serum was implemented on a Thermo Scientific™ Vanquish™ Flex Binary LC system connected to a Thermo Scientific™ TSQ Quantis™ triple quadrupole mass spectrometer. Detection was performed by selected reaction monitoring (SRM) using 25 deuterated internal standards. Method performance was evaluated using the ClinMass® TDM Platform with the ClinMass Add-On Set for Neuroleptics from RECIPE Chemicals + Instruments GmbH (Munich, Germany) to obtain limits of quantification, linearity ranges, accuracy, and intra- and inter-assay precision for each analyte.

Poster #15b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Various Other Development of an LC-MS/MS Method to Determine Incorporation Efficiency of Selected Modified Adenine Nucleotides into in vitro Transcribed RNA Dominika Strzelecka - Faculty of Physics, University of Warsaw *YI Grantee* ‣ The aim of the project is development of LC-MS/MS method to determine incorporation efficiency of selected modified adenine nucleotides into in vitro transcribed RNA. Modifications introduced within polyA tail may influence mRNA stability and yield transcripts with superior properties for the application in mRNA-based therapies. We used ion-pair chromatography coupled with ESI-QQQ to determine the concentration of adenine nucleotides obtained after RNA degradation and thereby determine RNA composition. The developed method is a tool for establishing mRNA structure-biological properties relationship.

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Poster #16f in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Various Other Tier 1 and Tier 2 Mass Spectrometry Methodologies for Determining Enzymatic Defects Associated with Krabbe Disease Collin Hill - PerkinElmer ‣ Krabbe disease is a rare lysosomal storage disorder caused by a defect in the galactosylceramidase (GALC) enzyme, rendering it unable to break down galactolipids which are abundant in the brain. It is believed that the accumulating galactolipids, namely psychosine, contribute to the destruction of myelinating oligodendrocytes, resulting in severe neurological disfunction. Therefore, it is crucial to detect Krabbe during the early days of life, during a critical development window for neonates. Here we describe two mass-spectrometry based assays which determine GALC activity through first tier rapid screening and subsequent confirmatory analysis of psychosine using LC-MS/MS in contrived positive samples.

Poster #17b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Various Other A Single High-Throughput UPLC-MS/MS Platform for Targeted Metabolomic, Lipidomic and Proteomic Studies (Targeted Multi-OMICS) Billy Molloy - Waters Corporation ‣ Targeted liquid chromatography - mass spectrometry (LC-MS) assays, using unit mass resolution, tandem quadrupole mass spectrometers, and standard analytical flow UPLC, are increasingly being used in the discovery omics arena. This approach offers an alternative to the untargeted, high-resolution, approach traditionally applied in these types of studies, using time of flight instruments and micro/nano flow LC. These methods offer increased simplicity and throughput, while still offering the level of sensitivity and specificity required Here, this methodology is applied to metabolomic, lipidomic and proteomic preparations of a set of human plasma samples, using the same LC-MS platform. The ability to run 3 different analysis types on the same LC-MS platform facilitates the consecutive analysis of multiple sample sets from different sample preparations with virtually no down time.

Poster #20b in Exhibit Hall - attended for 1 hr on Wednesday starting at 15:30 Topic: Various Other Identification of Hemoglobinopathies and Thalassemias : A Prospective Study of Up to 10,000 Newborn Samples Thomas Wiesinger - ARCHIMED Life Science GmbH ‣ The early diagnosis of hemoglobin disorders such as sickle cell anemia (SCD) or Thalassemias become more and more important due to increasing carrier-frequencies worldwide (approx. 330.000) as well as in the European Union, especially in urban areas (e.g. Berlin 1 birth in 2.500 newborns).[1] Since the composition of hemoglobin (fetal to adult) is affected by the age of the patient, a reliable strategy, which can handle this challenge of nature, was established and are currently tested in an ongoing study ( 10.000 samples).

Poster #22e in Exhibit Hall - attended for 1 hr on Wednesday starting at 10:00 Topic: Various Other Trimethylamine-N-Oxide Status in Patients with Helicobacter Pylori Eradication Therapy Assessed with Mass Spectrometry Andreas Meinitzer - CIMCL University Hospital Graz ‣ A routine method for the determination of trimethylamine-N-oxide (TMAO)in human serum was developed and tested in patients with antibiotic therapy.

Poster #22h in Exhibit Hall - attended for 1 hr on Thursday starting at 12:30 Topic: Various Other A Rapid and Simultaneous Quantification of Anti-Epileptic Drugs with a Liquid Chromatography-Tandem Mass Spectrometry (LC MS/MS) Method Recep Genç - Synlab Ankara Laboratory ‣ We aimed to develop a rapid and sensitive LC-MS/MS method for the simultaneous quantification of anti-epileptic drugs (AEDs) in plasma samples. The intra-day and inter-day precisions at two different concentrations were about 1.8-2.2% and 2-2.8% for Gabapentin, 2.9-3.4% and 3.3-4.8% for Levetiracetam, 3.8-3.6% and 4.7-4.8% for Lamotrigine, 4-4.8% and 4.6-5.1% for 10-OH-carbamazepin, 3.4-3.8% and 4-4.8% for Topiramate and 2-3.9% and 2.6-4.6% for Oxcarbamezapin, respectively. LODs and LOQs were 0.12, 0.18, 0.19, 0.18, 0.14, 0.095 mg/L and 0.73, 1.82, 0.77, 1.45, 0.55, 0.38 mg/L for Gabapentin, Levetiracetam, Lamotrigine, 10-OH-carbamazepin, Topiramate and Oxcarbamezapin, respectively. We recommend the use of this sensitive and simple LC-MS/MS method in clinical laboratories.

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Presenter Index

A

Adam, Mina ..... 26, 83 Adamski, Jurek ...... 22 Adhikari, Khem 26, 64 Adler, Carrie .......... 94 Albantakis, Laura .. 26,

83 Alves, Atecla .......... 79 Andersen, Ida Bøgh

.................... 26, 89 Anjo, Sandra .... 26, 88 Antonelli, Giorgia .. 80 Auray-Blais,

Christiane ... 28, 65, 71

B

Bagdonaite, Ieva .... 49 Balogh, Gábor ........ 40 Barco, Sebastiano . 26,

96 Barré, Florian ... 26, 43 Baumgarten, Sigrid 91 Belluomo, Ilaria 26, 47 Bendt, Anne ..... 41, 59 Bergmann, Marianne

.......................... 99 Binscheck-Domaß,

Torsten .............. 92 Borren van, Marcel 79 Boutin, Michel ....... 52 Braun, Valentin ...... 78 Bruszel, Bella ......... 89 Brzhozovskiy,

Alexander .... 26, 87 Burla, Bo .......... 26, 59 Bystrom, Cory .. 29, 54

C

Cameron, Simon ... 50, 68

Cangemi, Giuliana . 92 Carroll, Frances ...... 94 Cave, Rory ....... 26, 86 Chaudhari, Ravindra

.......................... 66

Chetouane, Yasmine .................... 26, 68

Cobbaert, Christa .... 6, 42, 66

Colas, Eva ............... 72 Coll de la Rubia, Eva

.......................... 87 Collier, Timothy ..... 45 Cordovana, Miriam 86

D

Darebna, Petra ...... 68 Dayot, Fanny .......... 92 de Grazia, Ugo ....... 91 De Nardi, Claudio ... 89 De Nicolò, Amedeo 91 Dekker, Shosha 26, 84 Delafiori, Jeany 26, 84 Delahaye, Lisa .. 26, 96 Denver, Nina .... 26, 81 Dihazi, Gry Helene . 95 Drouin, Nicolas ...... 46

E

Eberl, Anita ............ 95 Edgington, Alan ..... 92

F

Fabregat-Cabello, Neus 26, 47, 78, 99

Farre-Segura, Jordi26, 60

Fauler, Günter ....... 85 Frankevich, Vladimir

.................... 43, 97 Fredolini, Claudia .. 26,

88

G

Gaudl, Alexander ... 41 Genç, Recep ......... 100 Geyer, Philipp ........ 88 Giorgio Bellomo,

Marco Bagnati ... 83 Goryńska, Paulina

Zofia ............ 26, 86 Graça, Gonçalo 26, 46 Grant, Russ ...... 69, 70

Gray, Nicola26, 52, 64 Greaves, Ronda .... 23,

30, 48 Grenga, Lucia .. 26, 50 Gucciardi, Antonina

.................... 26, 89

H

Hammond, Gareth 98 Hanrieder, Jörg 26, 43 Harborow, Charlotte

.................... 26, 79 Hill, Collin ............ 100 HIRANO, Ichiro ...... 95 Horro Pita, Catarina

.......................... 63 Hurst, Emma ... 26, 90

I

Innarella, Maria Rosaria .............. 90

Ísberg, Ólöf Gerdur .................... 26, 43

J

Jaffuel, Aurore ...... 95 Jaroch, Karol ... 26, 85 Jauffrit, Frédéric ... 26,

50

K

Kazanc, Emine . 26, 98 Keevil, Brian 6, 37, 48,

57 Kern, Katharina ..... 93 Kim, Dong-Hyun .... 52 Kim, Moon-Ju .. 26, 93 Kim, Youngsoo ...... 66 Kitsilovskaya, Natalia

.................... 26, 82 Klinke, Glynis ......... 71 Konoshita, Ryu ...... 78 Kostrzewa, Markus 74 Kozak, Marta ......... 79 Križi?, Ivana ........... 83 Kuzyk, Valeriia . 26, 56

L

Lageveen-Kammeijer, Guinevere S.M. 26, 56

Lauc, Gordon ......... 67 Letertre, Marine ... 26,

64 Liigand, Jaanus 26, 65 Livie, Craig ....... 26, 96 Loftus, Neil ............ 79

M

Madunic, Katarina 26, 49

Majer, Hansjoerg .. 96 Mamedov, Ilgar ..... 84 Mandal, Govinda .. 26,

86 Markin, Pavel .. 26, 85 Marshall, David 26, 44 Mathew, Elizabeth 26,

82 McDonnell, Liam .. 24,

55 McNaughton, Melissa

.................... 26, 96 Meinitzer, Andreas

........................ 100 Melnikov, Arsenty 26,

58 Memarian, Elham . 26,

49 Mesihää, Samuel .. 26,

93 Mezger, Stephanie 26,

97 Mezzullo, Marco .. 26,

44 Michno, Wojciech . 97 Millington, David .. 25,

70 Mischak, Harald ... 23,

42 Molloy, Billy ........ 100 Moore, Edward 7, 24,

62 Moran, Alan .... 26, 56

102

Moreau, Stephane 94 Mortier, Leen ........ 90

N

Nicolardi, Simone .. 74 Nicolas, Mouton ... 26,

50

O

Ohlson, Sten .......... 44 Olin, Magnus ......... 94 Oppermann,

Madalina ........... 99

P

Panesar, Harrypal.. 47 Pap, Ádám ....... 27, 87 Piga, Isabella ... 27, 60 Pirro, Martina . 27, 72 Porta Siegel, Tiffany

.......................... 61 Pratt, Mark ...... 27, 41 Prost, Jean-

Christophe ........ 60

R

Rajska, Magdalena 98 Rappold, Brian 29, 75

Reinicke, Madlen .. 27, 84

Rola, Rafal ............. 82 Ruhaak, Renee 54, 66

S

Salimova, Dinara .. 27, 85

Santoru, Maria Laura .................... 27, 40

Saruhan, Ercan ...... 80 Schmedes, Anne .... 98 Schoenenberger,

Bernhard ........... 82 Schuster, Carina .... 93 Schwamborn, Kristina

................ 7, 61, 73 Senard, Thomas ... 27,

81 Sertoglu, Erdim ..... 80 Shackleton, Cedric . 21 Shuford, Christopher

.......................... 51 Sickmann, Albert ... 54 Smith, Andrew 27, 61 Spacil, Zdenek 27, 71,

84 Sriboonvorakul,

Natthida ...... 27, 65

Streit, Frank ........... 93 Strzelecka, Dominika

.................... 27, 99 Stuchlíková, Eliška 27,

88

T

Thevis, Mario ... 24, 53 Thorsteinsdottir,

Unnur Arna . 27, 71 Tiphaine, Robin 27, 65 Tokareva, Alisa ...... 81 Torta, Federico 27, 40 Toth, Gabor ........... 72

V

Van de Plas, Raf . 7, 73 van den Heuvel,

Dennis ............... 90 van der Velpen, Vera

.................... 27, 46 van Duijl, Tirsa . 27, 87 Vanhara, Petr ........ 81 Varela Coelho, Ana

.................... 27, 74 Vestergaard, Martha

Kampp Nøhr 27, 82 Vidali, Matteo ........ 78

Vogeser, Michael ....7, 29, 30, 53, 59

W

Wardle, Robert 91, 99 Wehrli, Patrick . 27, 97 Whiley, Luke ... 27, 52,

58 Wiesinger, Thomas

....................... 100 Woodfield, Georgia

.................... 27, 47 Wuhrer, Manfred ...7,

22, 67

Y

Yanshole, Lyudmila .................... 27, 58

Yanshole, Vadim ...27, 64

Yoon, Sohee .......... 98

Z

Zelzer, Sieglinde .... 80

103

MAP: 1st Floor - Exhibit Hall

Exhibits & Posters as well as Coffee Breaks, Lunches and Receptions will be held on the 1st Floor.

104

MAP: Tracks and Floors Plenary Lectures will be held in Europa (not shown, 2nd Floor) and Mozart Hall. Scientific Sessions and Corporate Workshops will be held on the Ground Floor, Paracelsus (2nd), Trakl (3rd) and Doppler (4th).

Documents

of The Association for Mass Spectrometry · The Association for Mass Spectrometry: Applications to the Clinical Lab Salzburg, AUSTRIA September 9 - 13, 2018 Salzburg Congress Center