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MSACL 2015 US: Preliminary Conference Program

San Diego, CA • March 28 - April 1, 2015

With Thanks to Our Corporate Sponsors:
Thermo Agilent Bruker Shimadzu SCIEX IONICS

SATURDAY
5:45 AM
6:45 AM
YOGA
@ Nautilus 5
Energize yourself for the day! Yoga is a complimentary offering for all MSACL registrants.
A limited number of yoga mats will be provided.
6:30 AM
8:00 AM
BREAKFAST
@ Harbor Island Foyer
8:00 AM
10:00 AM

SHORT COURSES: SESSION 1
Getting Started with Quantitative LC-MS/MS in the Diagnostic Laboratory
Judy Stone, PhD & Lorin Bachmann, PhD
Level: 1-2 (Beginner - Intermediate)
Location: Seabreeze 		Breaking up with Excel: A Newbie's Introduction to the R Statistical Programming Language
Daniel Holmes, MD & Stephen Master, MD PhD
Level: 1-2 (Beginner - Intermediate)
Location: Harbor 3 		Clinical and Research Applications using Inductively-Coupled Plasma Mass Spectrometry
Frederick Strathmann, PhD & Carrie Haglock-Adler, MSFS
Level: 1-2 (Beginner - Intermediate)
Location: Executive Center 4 		Intro to Clinical MS Method Development
Robert Kobelski, PhD
Level: 1-2 (Beginner - Intermediate)
Location: Spinnaker
General Toxicology
Jeffery Moran, PhD
Level: 1 (Beginner)
Location: Executive 2 		Understanding and Optimization of LC-MS/MS to Develop Successful Methods for Identification and Quantitation in Complex Matrices
Robert D. Voyksner, PhD
Level: 2 (Intermediate)
Location: Marina 6 		How to Develop Robust Assays Faster Using Free Data Analysis Tools
Fred Lytle, PhD
Level: 2-3 (Intermediate - Advanced)
Location: Marina 5 		A Comprehensive Review of Clinical Mass Spectrometry Technology & Techniques, including Miniaturization
Jack Henion, PhD
Level: 2-3 (Intermediate - Advanced)
Location: Executive 1
Modern Sampling and Sample Preparation Technologies for Clinical Mass Spectrometry
Janusz Pawliszyn, PhD & Barabara Bojko, PhD
Level: 1-3 (Beginner - Advanced)
Location: Executive Center 3 		Introduction to Quantitative Proteomics
Mike MacCoss, PhD & Michael Bereman, PhD
Level: 1-2 (Beginner - Intermediate)
Location: Harbor 1 		Development and Validation of Quantitative LC-MS/MS Assays for Use in Clinical Diagnostics
Russell Grant, PhD & Brian Rappold
Level: 3 (Advanced)
Location: Harbor 2
10:00 AM
10:30 AM
COFFEE BREAK
@ Harbor Island Foyer
Take a break and get a coffee, juice, water and/or small snack.

Commune with colleagues or perhaps go for a short walk outside by the water to refresh for the next session.
10:30 AM
12:00 PM

SHORT COURSES: SESSION 2
12:00 AM
1:00 PM
LUNCH
@ Harbor Foyer
Short Course Registrants Only: Pick up a box lunch and take advantage of the tables outside on the lawn or follow-up on some lingering questions with the instructor in the classroom.
1:00 PM
2:15 PM

SHORT COURSES: SESSION 3
2:15 PM
2:45 PM
COFFEE BREAK
@ Harbor Island Foyer
Take a break and get a coffee, juice, water and/or small snack.

Commune with colleagues or perhaps go for a short walk outside by the water to refresh for the next session.
2:45 PM
4:30 PM

SHORT COURSES: SESSION 4
Ends at 4:30 PM or at discretion of Instructor.
6:00 PM
8:00 PM
PRIVATE EVENT:Travel Grantee Reception & Dinner
@ Shoreline Patio by the Pool
** This event is exclusively for Travel Grantees. **
Dinner and Reception from 6:00 - 8:00 PM.
Thermo Waters Cambride Isotope Labs
8:00 PM
10:00 PM
HOSPITALITY
@ Shoreline Patio by the Pool
Enjoy the San Diego evening down by the Marina with heaters and fire pits.
Drinks provided.
10:00 PMSATURDAY CLOSED

SUNDAY
5:45 AM
6:45 AM
YOGA
@ Nautilus 5
Energize yourself for the day! Yoga is a complimentary offering for all MSACL registrants.
A limited number of yoga mats will be provided.
6:30 AM
8:00 AM
BREAKFAST
@ Harbor Island Foyer
8:00 AM
10:00 AM

SHORT COURSES: SESSION 5
Getting Started with Quantitative LC-MS/MS in the Diagnostic Laboratory
Continued from Saturday
Judy Stone, PhD & Lorin Bachmann, PhD
Level: 1-2 (Beginner - Intermediate)
Location: Seabreeze 		Breaking up with Excel: A Newbie's Introduction to the R Statistical Programming Language
Continued from Saturday
Daniel Holmes, MD & Stephen Master, MD PhD
Level: 1-2 (Beginner - Intermediate)
Location: Harbor 3 		Mass Spectrometry Toxicology Applications: Method Development and Validation, Troubleshooting, Triaging Instrumentation, Tips, Tricks and Lessons Learned
Marilyn Huestis, PhD & Karl Scheidweiler, PhD
Level: 1-3 (Beginner - Advanced)
Location: Executive Center 2 		Preparing Manuscripts for Publication: Improving Your Chances for Success
Thomas Annesley, PhD
Level: 0 (General Interest)
Location: Executive 4
Metabolomics
Gary Siuzdak, PhD & Gary Patti, PhD
Level: 3 (Advanced)
Location: Executive Center 3 		Intro to Clinical MS Method Development
Continued from Saturday
Robert Kobelski, PhD
Level: 1-2 (Beginner - Intermediate)
Location: Spinnaker 		Understanding and Optimization of LC-MS/MS to Develop Successful Methods for Identification and Quantitation in Complex Matrices
Continued from Saturday
Robert D. Voyksner, PhD
Level: 2 (Intermediate)
Location: Marina 6 		How to Develop Robust Assays Faster Using Free Data Analysis Tools
Continued from Saturday
Fred Lytle, PhD
Level: 2-3 (Intermediate - Advanced)
Location: Marina 5
A Comprehensive Review of Clinical Mass Spectrometry Technology & Techniques, including Miniaturization
Continued from Saturday
Jack Henion, PhD
Level: 2-3 (Intermediate - Advanced)
Location: Executive 1 		Clinical Proteomics
Andy Hoofnagle, MD, PhD & Cory Bystrom, PhD
Level: 2-3 (Intermediate - Advanced)
Location: Harbor 1 		Development and Validation of Quantitative LC-MS/MS Assays for Use in Clinical Diagnostics
Continued from Saturday
Russell Grant, PhD & Brian Rappold
Level: 3 (Advanced)
Location: Harbor 2
10:00 AM
10:30 AM
COFFEE BREAK
@ Harbor Island Foyer
Take a break and get a coffee, juice, water and/or small snack.

Commune with colleagues or perhaps go for a short walk outside by the water to refresh for the next session.
10:30 AM
12:00 PM

SHORT COURSES: SESSION 6
12:00 PM
1:00 PM
LUNCH
@ Harbor Foyer
Short Course Registrants Only: Pick up a box lunch in Harbor Foyer.
Travel Grantees see below.

Travel Grantees, you will be served Lunch outside at the Shoreline patio.

See Below
12:00 PM
1:00 PM
PRIVATE EVENT: Travel Grantee Lunch
@ Shoreline Patio by the Pool
** This event is exclusively for Travel Grantees. **

Lunch from 12:00 - 1:00 PM.
Thermo Waters Cambride Isotope Labs

1:00 PM
2:15 PM

SHORT COURSES: SESSION 7
2:15 PM
2:45 PM
COFFEE BREAK
@ Harbor Island Foyer
Take a break and get a coffee, juice, water and/or small snack.
Commune with colleagues or perhaps go for a short walk outside by the water to refresh for the next session.

POSTER PRESENTERS: If you are presenting a poster today now is the time to put up your poster.
2:45 PM
4:30 PM

SHORT COURSES: SESSION 8
Ends at 4:30 PM or at discretion of Instructor.
4:15 PM
7:00 PM
OPENING RECEPTION
@ Exhibit Hall
Enjoy mingling with colleagues and Exhibitors.

Take time to explore the Posters.

Heavy Appetizers and Drinks to be provided.
Sponsored by:
Thermo

5:00 PM
6:00 PM
POSTERS
@ Exhibit Hall
ALL Posters Attended & Presented.
Recognition Award Series
Distinguished Contribution Award Lecture
@ Harbor Ballroom
Chair: Andy Hoofnagle & Russell Grant
7:00 PM
8:00 PM
Tandem Mass Spectrometry in Clinical Diagnostics, Newborn Screening and Targeted Metabolomics
David Millington
Duke Medicine, Department of Pediatrics

MSACL 2015 US - Distinguished Contribution Awardee

A personal account of a journey spanning more than 30 years of applied mass spectrometry in a clinical setting is summarized in this lecture. 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 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 certain essential amino acids, a novel multiplex assay was developed to screen newborns for over 30 inherited metabolic conditions with a single test. This concept subsequently became the basis of a targeted metabolomics platform that was used 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.
8:00 PM
10:00 PM
HOSPITALITY
@ Shoreline Patio by the Pool
Enjoy the San Diego evening down by the Marina with heaters and fire pits.
Drinks provided.
10:00 PMSUNDAY CLOSED

MONDAY
5:45 AM
6:45 AM
YOGA
@ Nautilus 5
Energize yourself for the day! Yoga is a complimentary offering for all MSACL registrants.
A limited number of yoga mats will be provided.
6:00 AM
8:00 AM
BREAKFAST
@ Harbor Island Foyer
Enjoy a light continental breakfast that tastes even better after morning Yoga or a run along the bay, or maybe it tastes just fine by itself.
6:00 AM
8:00 AM
PLACE POSTERS
@ Exhibit Hall
Posters for Monday must be placed by the end of breakfast (8 AM).
7:00 AM
8:00 AM

CORPORATE WORKSHOPS (AM)
Restek
Marina 6

Pain Assays Do Not Have to Be a Big Pain! Analyze 230 therapeutic and drugs of abuse compounds by LC-MS/MS with the Restek Raptor Biphenyl
Frances Carroll, LC Application Chemist, Restek Corporation

The use of pain management drugs is steadily increasing. As a result, hospital and contract labs are seeing an increase in patient samples that must be screened for a wide variety of drugs to prevent drug abuse and to ensure patient safety and adherence to their medication regimen. The Raptor™ Biphenyl column was developed to complement high-throughput LC-MS/MS analyses. In this workshop, we will present the methodology for a 230 compound multi-class drug and metabolite screen and discuss the challenges one must consider when developing a large screening assay. Topics of the discussion will include mobile phase considerations, isobar resolution, drug interference, and Instrumentation. Optimized chromatography will also be presented for nine separate drug panels for use during confirmation and quantitative analyses.

Waters
Seabreeze

Towards Excellence in Steroid Hormone Measurements
Benjamin K. Beppler, TriCore Reference Laboratories, Development and Technology Scientist (1), Heather A. Brown, Ph.D , Senior Clinical Applications Scientist, Health Sciences Diagnostics, Waters Corporation (2)

This workshop will provide valuable details about the use of Waters’ Oasis µElution plate technology and column chemistries for the analysis of serum testosterone, androstenedione, and estradiol. These research methods have been evaluated for analytical sensitivity, reproducibility, and accuracy through participation in the Center for Disease Control and Prevention’s (CDC) Hormone Standardization (HoSt) Program. “Quantitation of 17â-Estradiol in Serum Using an Aggressive Sample Prep Method” “Quantitation of 17â-Estradiol in Serum Using an Aggressive Sample Prep Method”

Sigma-Aldrich
Spinnaker

Sample Preparation Considerations for Multiplexed MRM LC-MS Protein Assays in the Clinical Laboratory
Steve Hunsucker, Ph.D. - Senior Director of Laboratory Operations, Indi (Integrated Diagnostics)

Multiplexed protein assays have tremendous potential in clinical diagnostics, in particular measurement of proteins in plasma or serum derived from circulating blood. The dynamic range of protein concentration in these samples, and the dominance of very high abundance proteins such as albumin and immunoglobulins, make measurement of low concentration proteins impossible without some type of enrichment approach. This workshop will discuss the benefits of using Seppro® protein depletion columns in sample preparation for multiplexed LC-MS protein clinical assays.
8:10 AMWELCOME, INTRODUCTION & ORIENTATION
@ Harbor Ballroom
Welcome, Introduction and Orientation


In memory of Dr. Dr. Karl-Siegfried Boos


Update on MSACL EU.


Plans for next year and the future.


The Mobile Program App


BadgerScan Contact Collection: The next generation in contact and lead collection sharing. For vendors and attendees.
PLENARY LECTURE SERIES
@ Harbor Ballroom
Chair: David Herold
8:30 AM
9:15 AM
Phenotypings for Pharmacogenomics and Precision Medicine
Nigel Clarke
Quest Diagnostics
Precision medicine, also known as personalized medicine, is the term used for the model of health care involving the selection of diagnostic tests that have the potential to guide the physician towards the most efficacious course of treatment for the individual patient. The promise of precision medicine is a reduction in extraneous patient treatment with a concomitant increase in disease management efficacy based around the concept of “the correct drug for the correct reason”. This lecture will look at the role of precision medicine based around mass spectrometry and genetics; specifically the treatment of breast cancer with Tamoxifen and its metabolites. PK/PD relationships and their role in the use of pro-drugs such as Tamoxifen in the face of the various genetic mutations in the CYP family will be defined and the use of individual patient phenotypic metabolite measurements will be examined.
9:15 AM
10:00 AM
Innovations for Translating Metabolomics into the Clinical Laboratory
Rick Yost
University of Florida
Translating rapid advances in mass spectrometry and metabolomics into the clinical laboratory portends major changes in clinical analysis. Innovations in mass spectrometry, including ion mobility and FAIMS, ambient ionization, imaging mass spectrometry and global metabolomics all offer the opportunity for developing more sensitive, selective and rapid methods for the clinical laboratory. This lecture will include a perspective on the changing landscape for mass spectrometry in the clinical laboratory, insights into instrumental innovations, and examples of clinical applications.
10:00 AM
10:45 AM
COFFEE BREAK
@ Exhibit Hall
Visit the Exhibit Hall to procure coffee, juice, water and/or light snacks.

Explore what's on offer from the Exhibiting vendors, reconnect with colleagues, or go for a short walk outside by the water to refresh for the next session.

POSTER PRESENTERS: If you are presenting a poster today your poster should have been up 2 hours ago. If it is not up please put it up immediately.

GENERAL SCIENTIFIC SESSION 1
Track 1
Harbor Ballroom 1
Proteomics: Assay Quality and Performance
Chair: Cory Bystrom		Track 2
Harbor Ballroom 2
Renals & Adrenals
Chair: Julia Drees		Track 3
Harbor Ballroom 3
Metabolomics 1: Metabolic Reprogramming in Disease
Chair: Gary Patti		Track 4
Marina 6
Antimicrobial Susceptibility and Resistance Detection
Chair: Neil Anderson		Track 5
Executive Center
Basics of Mass Spectrometry
Chair: Rob Fitzgerald
10:45 AM
11:10 AM
Optimizing Trypsin Digestion
Kevin Meyer
Perfinity Biosciences
Long Abstract | Financial Disclosure | View Video
Digestion has historically been a significant bottleneck and source of irreproducibility associated with the preparation of certain protein samples for mass spectrometric analyses. This presentation will provide a comparison of the use of recombinant trypsin, pancreatic trypsin, additive/solvent enhanced digestion and elevated temperature methods as they apply to clinically relevant biomarkers including thyroglobulin and C-reactive protein as well as a highly multiplexed assay for apolipoproteins.		Measured GFR by Iohexol Clearance: A Pediatric Perspective
Shannon Haymond
Northwestern University Feinberg School of Medicin
Long Abstract
Accurate assessment of glomerular filtration rate (GFR) is critical for diagnosing and managing kidney disease and for safely prescribing and monitoring side effects of nephrotoxic and renally cleared agents. Estimated GFR (eGFR) is commonly calculated using equations based on creatinine and other parameters. Although relatively inexpensive and convenient, these equations are particularly problematic in children and adolescents. As efforts to refine and improve pediatric eGFR continue, there are frequently cases where an accurate method for mGFR is needed. We will describe our experience developing and implementing a semi-automated LC-MS/MS method for measurement of serum iohexol. Select case studies will be reviewed to demonstrate the clinical benefit of mGFR over eGFR in children. Details of the challenges faced in operationalizing an mGFR procedure will also be discussed.		Taming Tuberculosis: Metabolic Insights into the Host-pathogen Interface
Kyu Rhee
Weill Cornell Medical College
| Bio | View Video
Despite the advent of anti-infective chemotherapy well over 50 years ago, tuberculosis (TB) remains the bacterial cause of deaths worldwide. Unlike other pathogens, Mycobacterium tuberculosis (Mtb), the causative agent of TB, resides in humans as its only known reservoir and host. Mtb has thus evolved within an ultranarrow ecologic niche. Recent evidence has further highlighted metabolism as a key mediator of the host-pathogen interaction. Here, we present recent work highlighting specific elements of the bidirectional metabolic discourse between Mtb and the host. 		Rapid Antibiotic Susceptibility Detection Using Mass Spectrometric-Antibiotic Susceptibility Rapid Assay (MS-ASTRA) in Streptococcus Pneumoniae
Lawrence Tse

Medical College of Wisconsin
Long Abstract | Bio
In this study, we have adapted the Mass Spectrometric-Antibiotic Susceptibility Rapid Assay (MS-ASTRA) technique to determine penicillin resistance in over 50 clinical isolates of Streptococcus pneumoniae in less than 5 hours, compared to the overnight incubation requirement needed by many current methods. This technique uses MALDI-TOF to determine the relative amount of growth in the presence and absence of an antibiotic. Susceptibility data using MS-ASTRA was comparable to broth microdilution techniques (Phoenix, BD, Sparks, MD) with an overall agreement of over 90% and no very major errors.		Introduction to Mass Spectrometry
Jane Yang
UCSD
Long Abstract
Mass spectrometry is a powerful analytical chemistry tool that detects molecules in the gas phase based on their mass to charge ratio. This session will cover the basic concepts of mass spectrometry, how it works, the components of a mass spectrum, tandem MS (or MS/MS), the relationship between chromatograms and mass spectra, tuning, and what your service representative will do.
11:10 AM
11:35 AM
All that Glitters Is Not the Gold Standard: Calibration of a Sensitive Protein Cleavage-Isotope Dilution Mass Spectrometry (PC-IDMS) Assay for Thyroglobulin
Christopher Shuford
Laboratory Corporation of America
Long Abstract
A PC-IDMS assay was developed for quantification thyroglobulin in serum with a lower limit of quantification (LLOQ) of 0.2 ng/mL. To enable sensitive and accurate measurement at this LLOQ, external calibration was investigated with multiple sources of thyroglobulin and surrogate matrices lacking endogenous human thyroglobulin, including pooled human sera from remissive thyroidectomy patients (<0.1 ng/mL). Implications of matrix affects were explored as a function of thyroglobulin source and matrix, with consideration to the digestion process. Veracity of the final PC-IDMS assay was investigated by correlation with the FDA-approved Beckman Access® immunoassay and by comparison of two signature peptides.		Accuracy of Mass Spectrometry with the Ease of Immunoassay? LC-MS/MS Workflow Improvements for Primary Aldosteronism Screening
Daniel Holmes
University of British Columbia
Long Abstract | Bio | View Video
Uptake of LC-MS/MS is often hampered by perceived and actual workflow challenges and the enticement of walk-away automation offered by immunoassay. Both aldosterone and plasma renin activity (PRA) are difficult analytes with separate sample preparations by LC-MS/MS. We have significantly simplified the sample preparation of PRA and aldosterone to a single process. Comparisons with our current assays was excellent: Aldo-New = 0.99 × Aldo-Current + 10.1 pmol/L, R-sq=0.98 PRA-New = 0.94 × PRA-Current + 0.026 ng/mL/h, R-sq=0.99 Additionally, we present exploratory data on the aldosterone:AngI ratio as an equivalent screening tool to aldosterone:PRA ratios. 		Tracing Metabolism in Cancer
Matthew Vander Heiden
Koch Institute for Cancer Research at MIT
| Bio | Financial Disclosure
Cells adapt metabolism to meet distinct physiological needs, and metabolic regulation influences tumor progression. To proliferate, cancer cells must adapt metabolism to support anabolic processes and allow the accumulation of biomass. However, those nutrients with the highest consumption by cancer cells are not necessarily the fuels that contribute directly to cell mass. Cell culture provides a system to study how metabolism supports proliferation, but understanding non-proliferating cell populations requires an analysis of metabolism in patients and in tumor tissue. Use of mass spectrometry to trace nutrient use both in cell culture models and mouse cancer models will be presented to provide insight into how metabolism impacts cancer biology.		Quantitative Assessment of Multifactorial Resistance Mechanisms in Acinetobacter baumannii Using Selected Reaction Monitoring
Tiphaine Cecchini

bioMérieux
Long Abstract | Bio | Financial Disclosure | View Video
Multidrug resistant isolates of Acinetobacter baumannii responsible for nosocomial infections are reported increasingly. β-lactam resistance together with overexpression of RND efflux pumps are of particular concern. Following a simple sample preparation, a conventional liquid chromatography coupled with a triple quadrupole mass spectrometer was carried out in sMRM mode to detect in 1 hour both β-lactam hydrolyzing enzyme production (AmpC, OXA, TEM, GES, NDM, VIM, VEB, PER) and efflux pumps overexpression (AdeABC, AdeIJK) in multidrug resistant strains. Resistance phenotype might be deduced from these quantitative data. Correlation of mRNA and protein expression levels will be discussed for the efflux pump.		Introduction to Ionization Modes in Mass Spectrometry
Imir Metushi
University of California San Diego-CALM
Long Abstract | Bio | View Video
Ionization of analytes into the gas phase is a critical step for accurate mass identification. Various types of ionization methods can be applied for optimal analyte identification. This session will involve a basic introduction to various ionization modes used in mass spectrometry and will discuss the advantages and disadvantages of each.
11:35 AM
12:00 PM
Unraveling Trypsin Digestion, a Continuing Story: Mitigating Matrix Effects for Accurate MS-based Quantification of Serum Apolipoproteins
Irene van den Broek

Leiden University Medical Center (LUMC)
Long Abstract | Bio | Financial Disclosure | View Video
Despite excellent agreement and correlation between LC-MS/MS and immunoturbidimetric quantification of apolipoprotein A-I (apoA-I) in 100 patient sera, matrix effects on digestion completeness have been assigned as a remaining source of inaccuracy. In this study, we describe a detailed optimization of digestion conditions with a particular emphasis on reducing matrix effects on the digestion completeness of apoA-I, while additionally focusing on increasing peptide yield for multiple apolipoproteins (apoB-48, apoB-100, apoC-I, apoC-II, apoC-III, and apoE), and decreasing the digestion time. Effects of automation of the digestion protocol on assay simplicity, imprecision, and throughput will furthermore be discussed. 		The Benefits and Pitfalls of Using MRM3 Detection for the Analysis of Plasma Free Metanephrines by LC-MS/MS
Michael Wright
SEALS, Prince of Wales Hospital
Long Abstract | Bio | View Video
Liquid chromatography coupled to tandem mass spectrometry using multiple reaction monitoring (MRM) is a powerful tool for the quantitation of target analytes in complex matrices. However, this technique lacks specificity when plasma free metanephrines are measured. In this presentation we demonstrate the use of multistage fragmentation (MRM3) to improve the analytical selectivity of plasma free metanephrine service whilst also outlining other considerations that need to be taken into account before introducing this technology to a routine clinical chemistry laboratory.		Understanding the Metabolic Remodeling of the Heart Under Chronic Stresses
Rong Tian
University of Washington
Long Abstract | Bio | View Video
Energy metabolism is essential for maintaining normal cardiac function. Alterations of substrate metabolism, especially glucose and lipid metabolism; are integral to the development of pathological hypertrophy and heart failure. Furthermore, emerging evidence indicates that the branched-chain amino acids (BCAAs) catabolism is significantly changed during the development of cardiovascular and metabolic diseases. For example, plasma accumulation of BCAAs is strongly correlated with insulin resistance and coronary heart disease. Our recent studies describe a reciprocal regulation between glucose and BCAA metabolism in the heart. Investigation of the mechanisms governing the regulatory circuit will provide novel insight on the metabolic regulation of cardiac biology and diseases.		Solving a Microbiological Quandary: Does MALDI-TOF Mass Spectrometry Hold the Key to Rapid Detection of Vancomycin-intermediate Staphylococcus Aureus?
Susan Butler-Wu
University of Washington
Long Abstract | Bio
Vancomycin is a mainstay in the treatment of serious infections due to methicillin-resistant Staphylococcus aureus (MRSA). However, vancomycin treatment failure rates are higher in patients with blood-stream infection due to MRSA strains that have a vancomycin-intermediate sub-population (so-called heterogeneous VISA strains or “hVISA”). This subpopulation can often go undetected because it is present at a frequency lower than that what can be detected by standard susceptibility testing methods. In this study, we investigate the potential for MALDI-TOF Mass Spectrometry to rapidly detect VISA and hVISA strains. 		Introduction to Mass Analyzers: Quadrupole vs. Time-Of-Flight (TOF)
Alec Saitman

University of California, San Diego
Long Abstract | Bio | View Video
Choosing a mass analyzer for clinical analysis is an important step in setting up a mass spectrometry laboratory. But what is a mass analyzer? Are all mass analyzers are created equal? What types of clinical tests can be validated on each? Each type of mass analyzer has its own benefits and caveats in mass resolution, sensitivity, and dynamic range for small molecule analysis. Choosing a mass analyzer to suit the needs of the clinical laboratory is an important consideration to make. This lecture will focus on quadruple and time of flight (TOF) mass analyzers for identification and quantification of small molecules. This lecture will also describe how these different mass analyzers actually create mass separations and will describe the various clinical applications available to each.
12:00 PM
1:00 PM
LUNCH
@ Outside Harbor 3
Lunch to be provided in the Harbor & Bayview Foyers.

• Get ready to join a Corporate Workshop at 1:00 PM.

• Even-Numbered Posters to be Attended & Presented during the 2:00 - 3:00 PM coffee break in Exhibit Hall.

1:00 PM
2:00 PM

CORPORATE WORKSHOPS (PM)
Thomson Instrument Co
Harbor Ballroom 1

Improved Sample Preparation of Biological Samples using the Thomson eXtreme Filter Vials® and analysis by LC-MS/MS
Lisa Wanders, Technical Sales, Thomson Instrument Company

Sample preparation continues to be a critical factor in the quantitative measurement of biological samples. The goal of this seminar is to discuss how to streamline the sample preparation process of oral fluids and urine utilizing the Thomson eXtreme Filter VialsTM to reduce interferences from the sample matrix and increase analyte recovery. The Thomson eXtreme Filter VialsTM saves time, reduces solvent usage, alleviates the need for expensive consumables and lab equipment. Samples preparation for matrices such as urine and oral fluids will be discussed.

Thermo Scientific
Harbor Ballroom 2

Time Matters. Simplify Workflow Complexity.
Jason Lai Ph.D., MBA, Regulated Product Manager - Thermo Fisher Scientific

Pre-Register

Discover three newly listed Class I medical devices for general clinical use: Thermo Scientific™ Prelude MD™ HPLC, Thermo Scientific™ Endura MD™ mass spectrometer, and Thermo Scientific™ ClinQuan MD™ software. Clinical laboratories can use these devices to build their own lab developed tests (LDT). Combined, tools provide laboratories the ability to obtain the quantitative accuracy of LC-MS, easily and confidently. Examples of various compounds and workflows will demonstrate the robustness, stability, and time efficiency of these new class I medical devices.

Phenomenex
Harbor Ballroom 3

Solutions for Challenging Biological Matrices in Analytical Method Development
(1) Seyed Sadjadi, Ph.D., Phenomenex (2) Stephanie J. Marin, Ph.D., ARUP Institute for Clinical and Experimental Pathology

(1) When SPE is Not Enough: Considerations in developing a drug analysis in whole blood. We evaluate 7 common pretreatment procedures that hemolyze erythrocytes and precipitate plasma proteins from whole blood. The effectiveness of a procedure was based on overall recovery, response and reproducibility for analytes representative of different drug classes.

(2) Optimizing Sample Prep for Quantitation of Buprenorphine and Norbuprenorphine in Meconium. Meconium, the first stool produced by a newborn, is a complex matrix of materials ingested by a fetus, making it a good specimen to detect in utero drug exposure. Traditionally, meconium is homogenized in a laborious methodology. A simplified method where meconium is homogenized directly in enzyme, hydrolyzed and cleaned up by SPE to extract buprenorphine and norbuprenorphine is explored.

Bruker
Marina 6

(1) Robustness of the EVOQ Elite in Clinical Research Analysis
(2) MALDI-TOF MS in a Public Health Laboratory: Faster Bacterial Identification, Impact of Cost and Staff Savings and Future Applications
(3) Use of the Bruker Microflex™ LRF MALDI-TOF MS as a Rapid Screening Platform for Clinical Toxicology
(1) Dr. Timothy Garrett, Univ. of Florida, Gainesville, FL, (2) Kimberlee A. Musser, PhD, Wadsworth Center, NYSDOH, (3) Christina Wilson, PhD, Purdue University

(1) Due to the high sensitivity and selectivity provided by LC-MS/MS, this technology is being adopted in more clinical research applications. This talk will describe the unique design and performance characteristics of the EVOQ LC triple quadrupole system for the analysis of various research assays including Vitamin D, alcohol biomarkers, fructose and steroid profiling.
(2) The cost, staffing and turn-around time benefits with the adoption of the testing will be addressed. Additionally, validation of bacterial and mycobacterial identification and MALDI-TOF MS approaches to molecular serogrouping and assessment of antibiotic resistance will be discussed.
(3) This seminar will highlight the use of the Bruker Microflex™ LRF MALDI-TOF MS as a rapid, reliable tool for qualitative screening and confirmation of toxins and toxicants in clinical diagnostic toxicology cases.

2:00 PM
3:00 PM
COFFEE BREAK
@ Exhibit Hall
Visit the Exhibit Hall to procure coffee, juice, water and/or light snacks.

Explore what's on offer from the Exhibiting vendors, reconnect with colleagues, check out the posters, or go for a short walk outside by the water to refresh for the next session.
2:00 PM
3:00 PM
POSTERS
@ Exhibit Hall
Even-Numbered Posters to be Attended & Presented.
GENERAL SCIENTIFIC SESSION 2
Track 1
Harbor Ballroom 1
Proteomics: Endocrine Assays
Chair: Leigh Anderson		Track 2
Harbor Ballroom 2
Feeling your pain: TDM and Pain Medication Analysis
Chair: Katie Thoren		Track 3
Harbor Ballroom 3
Metabolomics 2: Metabolic Dynamics and Disease Pathogenesis
Chair: Kyu Rhee		Track 4
Marina 6
Beyond MALDI/Microbial Detection in Clinical Samples
Chair: Vanessa Phelan		Track 5
Executive Center
Tuning Your Mass Spectrometer
Chair: Rob Fitzgerald
3:00 PM
3:25 PM
High Sensitivity Measurement of Parathyroid Hormone Related Protein by LC-MS/MS for Diagnosing Disorders of Calcium Regulation
Mark Kushnir
ARUP Institute for Clinical and Experimental Path
Long Abstract | Bio
Measurement of Parathyroid Hormone related Protein (PTHrP) plays important role in follow-up of patients suspected of hypercalcemia. Concentrations of PTHrP in blood are very low and because commercial PTHrP immunoassays (IAs) are insufficiently sensitive, this result in a high false negative rate (FNR) of IAs and misdiagnosis of patients with elevated calcium. We developed LC-MS/MS method that overcomes drawbacks of commercial IAs and allows accurate measurement of PTHrP. Limit of quantitation of assay is 0.6 pmol/L, sensitivity sufficient to quantify PTHrP in samples of healthy and pathologic individuals. Our data suggest a FNR of one commercial IA of approximately 20%.		Formation of 6-acetylmorphine in Urine Samples with High Morphine Levels During Sample Preparation Involving Enzymatic Hydrolysis
Sihe Wang
Cleveland Clinic
Long Abstract | Financial Disclosure
6-Acetylmorphine (6-AM), an unique metabolite of heroin, is known as a definitive indicator of heroin intake. Due to variable glucuronide conjugation rates between and within individuals, an enzymatic hydrolysis using glucuronidase during sample preparation is frequently used to improve detection sensitivity and consistency. Acetate buffer is the primary choice for preparing enzymatic hydrolysis solution due to the desirable pH. We report here that urine samples with elevated levels of morphine (>100,000 ng/mL) incubated for >12 hours using an acetate buffer prior to LC-MS/MS analysis could form measurable amounts (¡Ý 5 ng/mL) of 6-AM.		Intestinal Fat Absorption and Systemic Metabolism: Applications of Mass Spectrometry in Discovery Research
Eric Yen
University of Wisconsin-Madison
Long Abstract | Bio | View Video
Triacylglycerol is the storage and transport molecule of fatty acids in animals. Its biosynthesis serves many physiological functions, including the absorption of dietary fat. However, excessive accumulation of triacylglycerol leads to obesity and related metabolic diseases. Acyl CoA:monoacylglycerol acyltransferase (MGAT) mediates one of the two TAG synthesis pathways. Among known MGATs, MGAT2 is highly expressed in the intestine of mice and humans. Findings from genetically engineered mice indicate that MGAT2 modulates the kinetics of fat absorption, the efficiency of energy metabolism, and the propensity to gain weight in response to calorie-dense diets. We are using proteomics as well as lipidomics profiling to explore molecular mechanisms underlying the functions of MGAT2.		Taxon-specific Markers for the Qualitative and Quantitative Detection of Bacteria in Human Samples
Nicole Strittmatter

Imperial College London
Long Abstract | Bio | Financial Disclosure
Based on a database of lipid profiles of microorganisms acquired using rapid evaporative ionization mass spectrometry, taxon-specific markers were derived for a variety of bacterial taxa at different levels. These markers were shown to be absent in human lipidome/metabolome and can thus be used to detect and quantify bacteria in human samples as exemplified by the imaging mass spectrometric analysis of human colorectal tissue. The approach can be used to detect certain types of bacteria in arbitrary matrices while maintaining both the untargeted and the spatially resolved nature of the mass spectrometric experiment.		Basics: Tuning Your Mass Spectrometer-How to Make It Sing!
Robert Fitzgerald
UCSD
Long Abstract | Bio | View Video
Tuning is fundamental to operating a mass spectrometer and involves mass calibration and mass resolution. This overview will focus on what happens when the mass spectrometer is tuned and introduces basic chemical concepts to help novice users understand what it means when your instrument representative says “I need to tune the mass spectrometer”.
3:25 PM
3:50 PM
A High-throughput Mass Spectrometry Multiplexed Assay to Measure Insulin and C-peptide
Steven Taylor
Quest Diagnostics Nichol's Institute
Long Abstract | Bio | Financial Disclosure
We have developed a multiplexed method to measure insulin and C-peptide using an LC tandem mass spectrometry assay. The assay involves enrichment of the peptides from patient sera using 2 different monoclonal antibodies immobilized on magnetic beads and processing on a Hamilton Star robotic liquid handler. Eluted peptides are directly analyzed by LC-MS/MS on an Agilent 6490 triple quadrupole mass spectrometer. The assay has a clinical reportable range from 2.5 to 320 µIU/mL for insulin and 0.11 to 27.2 ng/mL for C-peptide. Intra- and inter-day assay variation is less than 11% for both peptides.		Antiretroviral Testing: Development and Validation of LC-MS/MS Assays in Unique Specimen Sources to Support Clinical Trials
Mark Marzinke

Johns Hopkins University
Long Abstract | Bio
In order to better understand the pharmacokinetic-pharmacodynamic (PK-PD) relationships of antiretroviral drugs (ARVs) in disease prevention and management, compartmentalized PK studies are required to assess localized drug concentrations. Quantification of ARVs in cervicovaginal secretions (CVS) and rectal fluid (RF) can help determine efficacy at the site of viral transmission. Thus, liquid chromatographic-tandem mass spectrometric (LC-MS/MS) methods for the dual quantification of tenofovir and emtricitabine in CVS and RF have been validated according to the recommendations of the FDA, Guidance for Industry: Bioanalytical Method Validation document. Further, the described work illustrates the considerations for method validation in unique specimen sources.		Metabolomics by Masses, Metabolomics for the Masses
Nicola Zamboni
ETH Zurich
Long Abstract | Financial Disclosure | View Video
Metabolism plays a pivotal role in cellular processes by providing building blocks and energy for biosynthesis and participating in decision-making. In many biomedical areas such as nutrition, oncology, toxicology, stem cell research, etc., metabolism is currently regarded as a key driver and a differentiating factor to be exploited in diagnostics and selective therapy. To drive these activities, we developed a flow injection mass spectrometry platform that allows to routinely profile thousands of small molecules in biological extracts in thousands of samples per day. The massive throughput and information provided by our flow injection platform opens new fascinating horizons both in discovery and diagnostics.		Rapid Bacterial Identification Using a Mass Spectrometry Based Molecular Diagnostics Approach: Evaluation of the Iridica Platform
Alec Saitman

University of California, San Diego
Long Abstract | Bio | View Video
Abbott Diagnostics has developed the Iridica platform, capable of producing an accurate bacterial identification in less than 8 hours directly from blood specimens. The technology uses PCR based amplification of bacterial DNA followed by analysis by ESI-TOF mass spectrometry to make accurate identifications. The rapid identification is potentially useful in the clinical treatment and outcome of sepsis patients. In this study, we evaluated the clinical robustness of the Iridica platform by analyzing known positive blood cultures and outdated blood spiked with known bacterial isolates and demonstrated that the Iridica platform rapidly identified bacterial pathogens with good accuracy and precision in less than eight hours.		Basics: Compound Specific Tuning
Robert Fitzgerald
UCSD
Long Abstract | Bio | View Video
After ensuring that your instrument’s resolution and mass accuracy are set appropriately the next step in developing a quantitative LC/MS/MS assay based on multiple reaction monitoring (MRM) is to perform compound specific tuning. The established resolution and calibration files are used to optimize ion source electronics as well as gas flows for the compound of interest. This overview will focus on compound specific tuning and introduces basic concepts to help novice users understand what it means when your instrument representative says “We need to tune the mass spectrometer for your compound”.
3:50 PM
4:15 PM
Quantitation of Glycated Hemoglobin by MALDI Mass Spectrometry
Stephen Hattan
SimulTof Systems
Long Abstract | Bio | View Video
MALDI MS of whole blood hemolysates allows the direct quantitation of the total glycated hemoglobin (TGHb) ratios of alpha and beta chains from a single mass spectrum. HbA1c, the standard measure of glycated hemoglobin, can be calculated from TGHb. Sample preparation for this approach is minimal, analysis is rapid, 80 spots in 30 min (20 sec/spot), and hemoglobin variants are also detected. TGHb by MALDI vs. cation exchange HPLC, the reference standard, exhibited linearity (y = 0.79x + 1.4; r2 = 0.99) from 1.36% to 17.94% with CVs < 1.66%. MALDI method is better, faster, and less expensive than HPLC.		To Hydrolyze, or Not to Hydrolyze, that Is the Question for HRMS Pain Management Testing
Kara Lynch
University of California San Francisco
Long Abstract | Bio | Financial Disclosure | View Video
The demand for drug testing to monitor patients receiving treatment for chronic pain is continuing to increase. The objectives of this study were to 1) develop and validate a liquid chromatography high resolution mass spectrometry (LC-HRMS) screening panel for pain management testing and 2) evaluate two sample preparation approaches including hydrolysis and directly monitoring conjugated metabolites. HRMS offers a great platform for testing panels of compounds in one analytical run and shows good concordance with traditional MS based methods. Monitoring conjugated metabolites directly was sufficient for most analytes, however, there were 6 analytes missed in the 24 routine samples. 		Integrative Physiological Analysis of Adipocyte Metabolism
Christian Metallo
University of California, San Diego
| Bio
Obesity and metabolic syndrome are characterized by dysfunction in nutrient homeostasis, and adipose tissue can influence systemic metabolism via signaling or direct metabolic activity. Using a combination of oxygen physiology measurements and 13C metabolic flux analysis we have characterized how adipocyte hypoxia regulates amino acid metabolism. While branched chain amino acids (BCAAs) are normally major oxidative substrates for adipocytes, hypoxia elicits profound and lasting effects on glucose and amino acid metabolism such that BCAA catabolism is significantly compromised. These defects in adipose tissue oxygen physiology therefore contribute to the disturbances in amino acid homeostasis observed in the context of obesity.		MALDI Imaging Mass Spectrometry: Providing Molecular Insight at the Host-Pathogen Interface
Jessica Moore
Vanderbilt University
Long Abstract | Bio
Imaging Mass Spectrometry (IMS) provides specific molecular information directly from tissue while preserving spatial fidelity. As mass spectrometric technologies advance, high spatial (<10µm) and high mass resolution (>100,000 Resolving Power) IMS has been achieved. When applied to infected tissue, IMS can provide molecular information at the host-pathogen interface and molecularly define histological features without a priori knowledge of analytes. High mass resolution IMS using MALDI FTICR MS allows for isotopic resolution of protein species up to 10,000 MW and isolation of post-translationally modified epitopes in infectious lesions. Such analyses provide unprecedented capabilities for the study of infectious diseases.		Basics: Developing MRM Transitions
Robert Fitzgerald
UCSD
Long Abstract | Bio | View Video
After optimizing instrument tuning and compound specific tuning parameters the next step in developing a quantitative LC/MS/MS assay based on multiple reaction monitoring (MRM) is to identify appropriate fragment ions and optimize collision voltages for optimal sensitivity. This overview will focus on optimizing MRM transitions and introduces basic concepts to help novice users understand what it means when your instrument representative says “We need to develop MRM transitions for your assay”.
4:15 PM
7:00 PM
RECEPTION
@ Exhibit Hall
Enjoy mingling with colleagues and Exhibitors.

Take time to explore the Posters.

Heavy Appetizers and Drinks to be provided.
Sponsored by:
Thermo

5:00 PM
6:00 PM
POSTERS
@ Exhibit Hall
Odd-Numbered Posters to be Attended & Presented.

All posters to be removed by 7:30 PM.
7:00 PM
8:00 PM

DISCUSSION GROUPS
@ Harbor 1-3, Spinnaker & Marina 6
MSACL Jeopardy!
@ Harbor Ballroom 1
Lead(s): Robert Kobelski, Jack Henion & Jeff Moran

MSACL Jeopardy! A fun game of answer & question with topics covering aspects of clinical mass spectrometry. The audience will be divided into three teams who will race to pose questions to the answers provided in the jeopardy, double jeopardy and final jeopardy round.

Critical Method Development
@ Harbor Ballroom 2
Lead(s): Russell Grant & Brian Rappold

The discussion group will feature a short “Life in Review” of Professor Karl Siegfried Boos’s contributions to clinical diagnosis. This will be followed with a "Critical Method Review" competition – specifically highlighting sample preparation, a subject near and dear to Professor Boos.

Interest Group : Young Clinical Mass Spectrometrists
@ Harbor Ballroom 3
Lead(s): David Herold

The purpose of this discussion group is to create the opportunity for clinical mass spectrometrists, early in their careers, to assemble with the intent of creating and maintaining an interest group with a discernible voice that will communicate directly and effectively with MSACL, MSACL vendors and the clinical mass spectrometry community at large.

Regulations & Standards
@ Spinnaker
Lead(s): Julianne Botelho & Hubert Vesper

Open discussion on current regulations and standards for routine clinical laboratories employing mass spectrometry methods.

Vendor Feedback
@ Marina 6
Lead(s): Sharon McAvoy

Vendors invited to provide feedback regarding preferences for future MSACL conferences.
8:00 PM
10:00 PM
HOSPITALITY
@ Shoreline Patio by the Pool
Enjoy the San Diego evening down by the Marina with heaters and fire pits.
Drinks provided.
10:00 PMMONDAY CLOSED

TUESDAY
5:45 AM
6:45 AM
YOGA
@ Nautilus 5
Energize yourself for the day! Yoga is a complimentary offering for all MSACL registrants.
A limited number of yoga mats will be provided.
6:00 AM
8:15 AM
BREAKFAST
@ Harbor Island Foyer
Enjoy a light continental breakfast that tastes even better after morning Yoga or a run along the bay, or maybe it tastes just fine by itself.
6:00 AM
8:00 AM
PLACE POSTERS
@ Exhibit Hall
Poster for Tuesday must have their posters placed by the end of breakfast (8 AM).
7:00 AM
8:00 AM

CORPORATE WORKSHOPS (AM)
ZefSci
Seabreeze

Special Considerations for Clinical LC-MS/MS Method Validation
James Byrd, Application Chemist for Zef Scientific

Pre-Register

CLIA regulations require method validation, but do not specify which tests are required. We will talk about why LC-MS/MS methods may need extra validation steps compared to other high complexity tests.

Tecan
Spinnaker

Automation Solutions for Mass Spec Sample Preparation in the Clinical Laboratory
Daniel Leach, Senior Application Scientist

Although there have been monumental advances in mass spectrometry (MS) instrumentation in recent years, its unglamorous counterpart, sample preparation, has not enjoyed the same rate of development. This workshop will illustrate how the status quo is changing and will highlight a number of recent advances by Tecan utilizing its world class Freedom EVO liquid handling workstation to develop a variety of protocols designed to handle a broad range of sample types, sample preparation protocols and analytes to meet the needs of the modern analytical clinical laboratory. This includes both vacuum and positive pressure based SPE protocols, tip based cleanup for rapid processing of a small number of samples, sample tracking and LIMS integration.

IONICS MS
Spinnaker

Identification of Potential Biomarkers for Efficacy and Toxicity in Clinical Trials of a Cancer Vaccine using Targeted Metabolomic Profiling
Dr. Devanand Pinto

Pre-Register

Dr. Devanand Pinto will discuss what his Biomarker Quantification Team at the National Research Council of Canada has accomplished in their pursuit of a faster, more sensitive and more robust method of measuring immune response in cancer vaccine trials. Immune response is typically measured by cytokine profiling and flow cytometry; however, these techniques require extensive sample preparation and, due to the use of antibodies, have limited multiplexing capabilities. The Biomarker Quantification Team investigated the use of targeted metabolomics profiling to study the metabolic profile of 38 patients enrolled in a Phase I/Ib clinical trial for a novel cancer vaccine.This targeted metabolomic profiling approach utilizes an IONICS 3Q 320 which provides the leading sensitivity and advanced multiplexing capabilities necessary for success.
PLENARY LECTURE SERIES
@ Harbor Ballroom
Chair: Gary Siuzdak
8:30 AM
9:15 AM
Microfluidic Devices and Microscale Mass Spectrometry: Integration of Miniaturized Technologies for Acquiring Biological Information
J. Michael Ramsey
University of North Carolina at Chapel Hill
Mass spectrometry is a label free measurement technique that provides primary structure in addition to quantification, provided the target molecules can be placed into the gas phase as ions. While electrospray ionization has largely solved the problem of converting liquid-borne analytes into the gas phase ions, mass spectrometry has had the burden of being instrumentally complex, costly, and unwieldy.

The primary reason for the size, weight, and cost of mass spectrometers is the vacuum systems conventionally required for their operation. Over the past decade, we have been developing miniaturized mass analyzers that relax the necessary mean-free-path for operation. As a result these instruments can operate at pressures many orders of magnitude higher than conventional instruments, e.g., in the 1 Torr range. We refer to this mode of operation as High Pressure Mass Spectrometry (HPMS). HPMS allows mass spectrometry platforms that have size, weight, and power metrics that are all at least an order of magnitude smaller than the most compact conventional platforms. We have also been involved in integrating nano-electrospray (nESI) functionalities to microfluidic separation devices that yield state-of-the-art separations and nESI efficiency. The coupling of microfluidic nESI devices with HPMS platforms and their potential applications will be discussed in this presentation.

9:15 AM
10:00 AM
The Rise of Intelligent Machines and What It Means to the Lab and Healthcare
Randall Julian
Indigo BioAutomation
Long Abstract | Financial Disclosure
Intelligent machines teamed with experts are superior to experts working alone. This will have profound effects on the nature of healthcare delivery. Further, the advance of automation is already having a significant effect on labor markets, and there is no reason to believe healthcare will not be impacted. In this lecture examples of human-machine teams will be given. Also, the impact on society of the increased role of smart machines will be discussed. Comparisons between the first and second machine ages will be used to draw out the consequences, benefits and difficulties we will face as a scientific community.
10:00 AM
10:45 AM
COFFEE BREAK
@ Exhibit Hall
Visit the Exhibit Hall to procure coffee, juice, water and/or light snacks.

Explore what's on offer from the Exhibiting vendors, reconnect with colleagues, or go for a short walk outside by the water to refresh for the next session.

POSTER PRESENTERS: If you are presenting a poster today your poster should have been up 2 hours ago. If it is not up please put it up immediately.
GENERAL SCIENTIFIC SESSION 3
Track 1
Harbor Ballroom 1
Proteomics: Methods and Strategies
Chair: Tim Collier		Track 2
Harbor Ballroom 2
Small Molecule Analysis: Sunshine and Datamine
Chair: Daniel Holmes		Track 3
Harbor Ballroom 3
Metabolomics 3: Pathways to Lipids
Chair: Mohit Jain		Track 4
Marina 6
Microbial Metabolomics
Chair: Pieter Dorrestein		Track 5
Executive Center
Fundamentals: Sample Prep
Chair: Judy Stone
10:45 AM
11:10 AM
Development of Clinical Assays Based on Parallel Reaction Monitoring
Bruno Domon
Luxembourg Clinical Proteomics Center
Long Abstract | Bio | View Video
Targeted analyses of clinical samples performed on a quadrupole-orbitrap instrument using parallel reaction monitoring showed a significant gain in sensitivity and selectivity. In order to fully leverage the potential of this approach to develop clinical assays, we have designed a new data acquisition scheme. It relies on added internal standards and on-the-fly adjustment of acquisition parameters to drive in real-time the measurement of endogenous peptides (corresponding to proteins of interest) and generate precise and high confidence quantitative results. Applied to the analysis of lung cancer markers in plasma samples, it improved the discrimination of the disease stages and subtypes. 		Comparison of CDC’s Candidate Reference Method for Serum 25-hydroxyvitamin D3 and 25-hydroxyvitamin D2 with Recognized Reference Methods
Ekaterina Mineva
CDC
Long Abstract
Vitamin D status is routinely assessed by measuring serum concentrations of the most stable and abundant vitamin D metabolites, namely, 25-hydroxyvitamin D3 and 25-hydroxyvitamin D2. To assure the accuracy of measurements, worldwide vitamin D standardization activities are ongoing. As part of these efforts, we have developed a candidate reference measurement procedure to support CDC’s Vitamin D Standardization Certification Program. We are presenting a comparison of the serum concentrations of 25-hydroxyvitamin D3 and 25-hydroxyvitamin D2, measured using CDC’s candidate reference method, to target concentrations established by JCTLM-recognized reference methods (University of Ghent and NIST) in various single donor samples and standard reference materials. We will compare method features and highlight performance. 		Discovery and Characterization of Novel Bioactive Mammalian Lipids
Alan Saghatelian
The Salk Institute
Long Abstract | Bio | View Video
Lipidomics led to the identification of a novel class of mammalian lipids which have beneficial metabolic effects – Branched Fatty Acid esters of Hydroxy-Fatty Acids (FAHFAs). A member of this class, Palmitic Acid Hydroxy Stearic Acid (PAHSA) has at least 8 isomers which are present in serum and many mouse and human tissues. And administration of these lipids to mice improved various metabolic parameters. These lipids are of interest as biomarkers and potential therapeutics. 		Three-dimensional Metabolic Imaging of Live Bacterial Colonies by Laser Ablation Electrospray Ionization Mass Spectrometry with Ion Mobility Separation
Hang Li

George Washington University
Long Abstract | Bio | View Video
With the rising tide of antibiotic resistant bacterial strains, understanding microbial metabolism is of great importance. Conventional bioanalytical tools often have limited ability to provide spatiotemporal metabolite distributions in microbial colonies. Here, we introduce laser ablation electrospray ionization (LAESI), in combination with ion mobility separation (IMS) and mass spectrometry (MS), to rapidly investigate microbial metabolism. Over one hundred metabolites and lipids were detected by LAESI-IMS-MS from a single bacterial colony within seconds. Three-dimensional images were built from lateral rastering and depth profiling. For example, the m/z 188.2 ion, identified as acetylspermidine, exhibited stronger signal on the perimeter of the colony. Our results show that high throughput metabolic analysis and three-dimensional imaging can be performed by LAESI-IMS-MS. 		Part 1: Short Review of LC-MS/MS Sample Preparation Principles
Julia Drees
Kaiser Permanente Regional Laboratory
Long Abstract | Bio | View Video
This presentation is an introduction prior to an in-depth case study examining sample preparation. The most common types of sample clean-up used in clinical LC-MS/MS will be discussed along with their relative strengths and weaknesses. Ion suppression will be introduced and a robust method to quantify matrix effects and recovery will be explained. Options for automating sample prep will be presented.
11:10 AM
11:35 AM
Evaluation of Different Mass Spectrometry Data Acquisition and Analysis Strategies for Amyloidosis Typing
Han-Yin Yang

University of Washington
Long Abstract | Bio | View Video
Laser microdissection couple with mass spectrometry (LDMS) based amyloidosis diagnosis method has been a better alternative to immunohistochemistry method. However, several aspects need to be considered in the assessment of a reliable diagnosis platform. Here, we apply different normalization approach to correct for the variances in LD capture quantities and sample preparation. In addition, we quantify the difference between samples using peptide and protein levels, and discuss the diagnosis ability of different quantification approaches. Preliminary results show data-independent acquisition followed by intensity based method provides more evidence for diagnosis.		Direct Quantitation via Internal Standard Ratios for LC-MS/MS Assays
Brian Rappold
Essential Testing
Long Abstract | View Video
Diagnostic MS/MS methods largely utilize calibration curve-based quantitation on a batch basis. Initial forays into internal standard ratio quantitation have been recently proposed. This mode allows for smaller batches, faster run-times and expedited data review while still leveraging the analytical horsepower of mass spectrometric platforms. Additionally, this workflow satisfies the core requirements of quality control under CLIA and various accrediting agencies. These approaches, however, do not address non-linear MS detection, increased imprecision at low responses or noise/baseline variations between analytes and internal standard transitions. This paper shall introduce a highly accurate means of internal standard ratio quantitation via calculation of correction factors across the entire analytical range for a diverse array of analytes and internal standard labeling conditions.		Metabolomic Analyses of Subclinical Indicators and Predictors of Disease
Clary Clish
Broad Institute
Long Abstract | Bio | View Video
We have developed a robust LC-MS-based metabolomics platform that is capable of measuring hundreds of intermediate metabolites in multiple pathways, including glucose and lipid metabolism, and we have applied this technology to profile samples derived from cell culture, body fluids, tissues, and microbes. We have investigated whether plasma metabolite profiles predict future development of diabetes in participants of the Framingham Heart Study. These analyses revealed amino acid and lipid signatures that are predictive of risk and were replicated in an independent, prospective cohort. In addition to these results, findings from recent efforts to find metabolic indicators of pancreatic cancer in will be presented.		Harnessing Metabolomics for Microbial Identification in Complex Samples
Vanessa Phelan
University of California, San Diego
Long Abstract | View Video
Current mass spectrometry methods in clinical laboratories for microbial detection are limited to identifying the genus and species. However, it is well established that microbes produce a number for organism specific metabolites to interact with their environments. These metabolites function as toxins, antibiotics, redox active molecules and nutrient scavenging entities. Using newly developed bioinformatics tools, we can begin to evaluate whether specific specialized metabolites are detectable in clinical samples and if those metabolites can provide insight into patient status. Here, we describe the application of MS/MS based molecular networking to identify Pseudomonas aeruginosa metabolites in sputum samples from CF patients.		Part 2a: The Trials and Tribulations of Sample Preparation for Testosterone by LC-MS/MS
Deborah French
University of California, San Francisco
Long Abstract | Bio | View Video
Use of liquid chromatography-mass spectrometry for analysis of small molecules in clinical laboratories has increased in the past few years. An important component of developing a mass spectrometry method is the sample preparation technique employed in order to sufficiently clean up the sample without sacrificing analyte recovery. A number of references out there detail sample preparation techniques but which one do you choose? This presentation will detail the thought process behind the sample preparation technique choice for measurement of serum total testosterone in one laboratory, including the balancing act of what you want to, and what you can realistically achieve.
11:35 AM
12:00 PM
Development of an Automated Exosome Isolation Procedure for Determining Prostate Cancer Aggressiveness
Alex Rai
Columbia University Medical Center
Long Abstract | Bio | Financial Disclosure | View Video
Almost 230,000 men are diagnosed with prostate cancer annually, but over 900,000 men undergo biopsy. A biomarker to identify aggressive disease could reduce biopsies and associated morbidity. Exosomes are small (30-200 nm) membranous vesicles secreted by all cells. They harbor biomarkers and can serve as a real-time “liquid biopsy”. Proteomics analysis using 1D LC-MS/MS and multidimensional protein identification technology (MudPIT) was performed after trypsin digest. A 44 exosomal-protein signature was delineated to distinguish patients with metastatic disease and those after radical prostactectomy. The 44 proteins can be classified into eight major functions, identifying pathways involved in disease progression & metastasis.		Using Database Mining on Residual Samples to Establish Healthy Reference Intervals for Testosterone Measured by LC-MS/MS
Julia Drees
Kaiser Permanente Regional Laboratory
Long Abstract | Bio | View Video
Establishing reference intervals is challenging for clinical laboratories. In the case of testosterone, clinical laboratories must decide whether to consider the age-related decline in male testosterone a normal phenomenon which should be reflected in the reference intervals. In this study, automated electronic medical record database mining was used to identify residual samples from healthy adult males and females for use in establishing total and calculated free testosterone reference intervals. Total testosterone was measured on an LC-MS/MS assay that demonstrated excellent correlation with the CDC reference method. Data from 292 adult males did not support decreasing reference intervals for total testosterone with increasing age. In contrast, free testosterone was observed to decline.		Credentialing Biologically Relevant Features: Honey, I Shrunk the Metabolome
Gary Patti
Washington University
In LC/MS-based metabolomics, it is routine to detect thousands of features from the metabolic extract of a biological sample. When the mass-to-charge values of these features are searched exhaustively in available databases, however, about 50% do not return hits. This raises questions about the size of the metabolome and the number of potential unknown structures that have yet to be characterized. We have developed a platform called "credentialing" to identify features in a metabolomic experiment that are of biological origin. In brief, unlabeled and labeled samples are mixed at unique ratios and analyzed together on the basis of isotope spacing and intensity. The credentialing platform enables data reduction and improved identification of bona fide unknowns. The application of credentialing to identify an unknown altered in cancer cells will be discussed.		Direct Microbial Analysis by Ambient Ionization MS: Paper Spray and Swab Touch Spray
Alan Jarmusch

Purdue University
Long Abstract | Bio | View Video
Ambient ionization mass spectrometry allows for rapid analysis of bacteria and fungi samples with minimal sample preparation. The biomolecule profiles (e.g. lipids) detected are characteristic and allow for molecular-based identification via multivariate statistics. Paper spray (PS) ionization allowed differentiation of various species of bacteria and yeast, requiring only a minute amount of material applied to a paper triangle. Swab touch spray (STS), a novel method developed for in vivo sampling, allowed direct microbial analysis from swabs. PS and STS methods for microorganism identification can extend into clinical medicine, enhancing current analytical methods with significant impact on patient treatment.		Part 2b: The Trials and Tribulations of Sample Preparation for Testosterone by LC-MS/MS, CONTINUED
Deborah French
University of California, San Francisco
Long Abstract | Bio | View Video
Use of liquid chromatography-mass spectrometry for analysis of small molecules in clinical laboratories has increased in the past few years. An important component of developing a mass spectrometry method is the sample preparation technique employed in order to sufficiently clean up the sample without sacrificing analyte recovery. A number of references out there detail sample preparation techniques but which one do you choose? This presentation will detail the thought process behind the sample preparation technique choice for measurement of serum total testosterone in one laboratory, including the balancing act of what you want to, and what you can realistically achieve.
12:00 PM
1:00 PM
LUNCH
@ Outside Harbor 3
Lunch to be provided in the Harbor & Bayview Foyers.

• Get ready to join a Corporate Workshop at 1:00 PM.

• Even-Numbered Posters to be Attended & Presented during the 2:00 - 3:00 PM coffee break in Exhibit Hall.

1:00 PM
2:00 PM

CORPORATE WORKSHOPS (PM)
Waters
Harbor Ballroom 1

Clinical Research and Biomarker Discoveries, Challenges and Opportunities
(1) Dr. Amrita K Cheema, Associate Professor & Co-Director-PMSR, Georgetown University Medical Center. (2) J. Will Thompson, Ph.D., Assistant Research Professor, Proteomics and Metabolomics, Duke University

In this workshop, the directors of the large MS core facilities at Duke and Georgetown University Medical Centers will discuss the real-life challenges of, and the opportunities for conducting clinical research and biomarker discovery using state-of-the-art MS-based approaches, including proteomics, lipidomics and metabolomics. Application examples, useful tips and tricks on how to manage large MS experiments, and some practical MS troubleshooting advice will be provided. Presentation #1: Supporting the Systems Medicine Paradigm: Metabolomics and Lipidomics Core Technologies for Clinical and Translational Research Presentation #2: The Utilization of Quality Measures in Experimental Design and Interpretation to Aid Quantitative Clinical Proteomics and Metabolomics Studies”

Shimadzu
Harbor Ballroom 2

Secrets to Successful LC-Tandem MS Implementation - Tips and tricks for Triple Quadrupole Mass Spec Techniques
Kent Johnson (Exceltox Laboratories, LLC), Chris Gilles (Shimadzu Scientific Instruments)

From screening to confirmation/quantitative testing, there are many pitfalls that can prevent maximum laboratory efficiency. This workshop will discuss several specific scenarios and offer solutions to everyday challenges in medication monitoring and urine drug testing, including: - Better sample preparation, - Validity Testing (to ensure the sample is human and that it has not been diluted nor adulterated), - Detection of early signs of medication misuse or illicit drug use, - State of the art LC/MS technology, - Delivering quality data on an ultra-fast timescale, and - Simplifying reporting and billing. Please join us for this practical workshop and learn how to operate your laboratory at maximum efficiency. Attendees will receive a lunch, t-shirt and GiantMicrobe!

Agilent Technologies
Harbor Ballroom 3

(1) Development of a Multiplexed MRM LCMS LDT
(2) Challenges in Quantitative Dried Spot Sampling and Analyses
(1) Stephen W. Hunsucker, Ph.D., Integrated Diagnostics (2) Kenneth Lewis, Ph.D., OpAns, LLC

Pre-Register

(1) The potential impact of multiplexed MRM LCMS analysis for clinical diagnostics is enormous and has garnered much attention in recent years. This workshop will discuss the approach for the development of multiplexed MRM LCMS LDT(s) and highlight some of the analytical challenges associated with the design and development of such an LDT.

(2)The Pediatric Trial Network (PTN) was established to create an infrastructure for investigators to conduct trials that improve child health. Advancement of low volume sampling technologies is being achieved through these trials by demonstrating the concordance of drug concentrations in plasma to Dried Blood Spots (DBS). In this presentation, Dr. Lewis will give a brief description of challenges and remedies encountered in the quantitative analysis of dried spot specimens for small molecule targets.

Biotage
Marina 6

Advances in Sample Prep for Pain Management: Developing a Simple, Fast Method for Nonpolar Compounds and Polar Metabolites in Urine prior to LC/MS
Matthew Slawson, PhD

Described here is a method utilizing supported liquid extraction for the detection of tapentadol, its glucuronidated, sulfated, and N-desmethyl metabolites; tramadol, its N- and O- desmethyl metabolites; and meperidine and its N-desmethyl metabolite. The method utilizes ISOLUTE SLE+ (Biotage; Charlotte, NC) for sample clean-up followed by analysis by LC-MS/MS. The extraction was optimized to ensure good recovery of both nonpolar parent drug polar metabolites by utilizing a sample pretreatment with a basified brine solution and elution with acidified methylene chloride/isopropanol. These steps ensured that both polar and nonpolar analytes eluted in the same extract. The method has a limit of detection of at least 50 ng/mL and an upper limit of linearity of at least 5,000 ng/mL. The method is simple, fast and offers excellent S:N.
2:00 PM
3:00 PM
COFFEE BREAK
@ Exhibit Hall
Visit the Exhibit Hall to procure coffee, juice, water and/or light snacks.

Explore what's on offer from the Exhibiting vendors, reconnect with colleagues, or go for a short walk outside by the water to refresh for the next session.
2:00 PM
3:00 PM
POSTERS
@ Exhibit Hall
Even-Numbered Posters to be Attended & Presented.
GENERAL SCIENTIFIC SESSION 4
Track 1
Harbor Ballroom 1
Proteomics: Clinical Assays I
Chair: Stephen Master		Track 2
Harbor Ballroom 2
Optimization of HRMS
Chair: Kara Lynch		Track 3
Harbor Ballroom 3
Metabolomics 4: High-Throughput & Computational Approaches to Lab Medicine
Chair: Caroline Johnson		Track 4
Marina 6
Diagnostic Gaps in Infectious Disease Testing
Chair: Nate Ledeboer		Track 5
Executive Center
LC Separation
Chair: Judy Stone
3:00 PM
3:25 PM
Detection of CSF Proteins Using LC-MS/MS: Measurement of Beta-Trace Protein as an Indicator of a CNS Breach
Kari Gurtner
Mayo Clinic
Long Abstract | Bio | View Video
The presence of CSF in a body fluid, indicative of a CNS breach, is currently detected by gel immunofixation (IFE) and nephelometric quantification. Both methods rely on the detection of a single marker for CSF and have limitations in the presence of serum contamination. Testing for beta-trace proteotypic peptides by LC-MS/MS aims to overcome those limitations. Of 104 body fluids tested, 96% of the samples were in agreement between IFE and LC-MS/MS testing. Peptidyl analysis by LC-MS/MS increases the accuracy of detection of CSF proteins without being impacted by non-CSF contamination or bacterial deglycosylation, offering superior sensitivity and specificity.		Broad Spectrum Drug Screening Using Liquid Chromatography Quadrupole Time-Of-Flight Mass Spectrometry: How Valuable Is Retention Time for Identifying Compounds?
Katie Thoren

Memorial Sloan Kettering Cancer Center
Long Abstract | Bio | View Video
Using a LC-QTOF mass spectrometer for broad-spectrum drug screening, compounds are identified based on their precursor mass, isotope pattern, retention time and product ion spectra. Ideally, assays would only be based on intrinsic compound parameters so that changes in method conditions would not affect compound identification. Out of these four parameters, retention time information is the most method-dependent. But how valuable is it for compound identification, especially when product ion spectra are available? Using 100 routine clinical urine samples, we compared how well our method identifies compounds with and without the use of retention time information. Ultimately, our goal is to make drug screening methods more universal while still achieving reasonable compound identification.		Metabostasis of the Aging Brain
Gary Siuzdak
The Scripps Research Institute
| Bio
Brain structure and function are highly dependent upon metabolic homeostasis (metabostasis) however it is unknown how this is affected during the natural aging process. Biochemical characterization of metabostasis would provide important insight into brain metabolism and potential impairment related to metabolic dysfunction as a hallmark of aging. Here we apply cutting-edge, mass spectrometry-based metabolomics to characterize metabolites across anatomical regions of a mouse brain at different stages of the life cycle. This temporal overview of metabostasis integrated with protein expression data and metabolite magnetic resonance imaging further validated and help define regional brain metabolism during the healthy aging process. 		PANEL: Diagnostic Gaps in Infectious Diseases: a Proposal for an Interdisciplinary Approach to Develop Mass Spectrometry Methods to Find the Bug and Treat the Host
Carey-Ann Burnham Et Al. (see Long Abstract)
Washington University
Long Abstract
The application of Mass Spectrometry in the clinical microbiology laboratory has been limited mainly to the identification of microorganisms growing in culture. However, there are a large number of technological gaps in both the diagnosis and management of infections that could potentially make ideal candidates for MS-based solutions. In this panel discussion, three clinical microbiologists will present clinical problems and/or unmet diagnostic needs in the field of clinical microbiology. A panel of experts in MS will then respond to each need presented, weighing in on the potential feasibility of a MS-based analytical solution. The major objective of this session is to foster interdisciplinary collaborations to improve both the diagnosis and management of infections using novel MS-based assays.		Part 1: Short Review of LC Method Development Principles
Julia Drees
Kaiser Permanente Regional Laboratory
Long Abstract | Bio | View Video
This presentation will introduce the principles of reverse-phase and HILIC high performance liquid chromatography (HPLC) method development and address common problems such as column overload, injection matrix/mobile phase mismatch, and poor ionization. Best practices and maintenance suggestions will be discussed.
3:25 PM
3:50 PM
Using MALDI-TOF MS to Screen for Monoclonal Proteins in Serum
Mindy Kohlhagen
Mayo Clinic
Long Abstract | Bio | View Video

Replacing Robert Bergen

This study aims to evaluate MALDI-TOF MS as a clinical screening method for serum monoclonal proteins (M-proteins). A set of 556 serum samples previously tested by gel electrophoresis (PEL) and immunofixation (IFE) were analyzed by MALDI-TOF MS. The mass distributions of immunoglobulin light chains from each sample were compared to normal serum. Deviations from the normal distribution were considered positive. Upon analysis, 100% of PEL positive, 91% of IFE positive and 19% of PEL/IFE negative samples screened positive by MALDI-TOF MS. The results suggest that MALDI-TOF MS has near equivalent sensitivity compared to current methods and would be an effective screen for M-proteins.		Comparing TOF and QTOF for Comprehensive Drug Screening: Do You Really Need Fragmentation Information?
Jennifer Colby

University of California San Francisco
Long Abstract | Bio | Financial Disclosure | View Video
High resolution mass spectrometry (HRMS) is an emerging technique that has been applied to comprehensive drug screening. HRMS instruments, including time of flight (TOF) analyzers, measure mass accurately. Quadrupole TOF (QTOF) mass analyzers can also select precursor ions and produce compound specific fragmentation patterns. We compared the ability of a QTOF instrument to identify drugs in patient samples, including and excluding fragmentation information. We found that including fragmentation patterns improved the assay’s sensitivity by 11% and improved positive predictive value by 25%. Fragmentation patterns increase confidence in compound identification and may allow screening results to be released without confirmatory testing.		High Throughput Monitoring of Small Molecules in Human Plasma
Mohit Jain
University of California, San Diego
Both internal and external environmental factors are critical modulators of human disease through the introduction of small molecules into circulating plasma. Traditional LC-MS based approaches for assessing plasma small molecules are limited in their throughput. In our talk, we will introduce new approaches for high throughput measure of plasma small molecules using automated in-line SPE with direct infusion mass spectrometry, and the association of small molecules with human disease. 		PANEL: Diagnostic Gaps in Infectious Diseases: A Proposal for an Interdisciplinary Approach to Develop Mass Spectrometry Methods to Find the Bug and Treat the Host
Susan Butler-Wu Et Al. (see Long Abstract)
University of Washington
Long Abstract
The application of Mass Spectrometry in the clinical microbiology laboratory has been limited mainly to the identification of microorganisms growing in culture. However, there are a large number of technological gaps in both the diagnosis and management of infections that could potentially make ideal candidates for MS-based solutions. In this panel discussion, three clinical microbiologists will present clinical problems and/or unmet diagnostic needs in the field of clinical microbiology. A panel of experts in MS will then respond to each need presented, weighing in on the potential feasibility of a MS-based analytical solution. The major objective of this session is to foster interdisciplinary collaborations to improve both the diagnosis and management of infections using novel MS-based assays.		Part 2a: Serum Aldosterone: An LC Method Development Case Study for a Difficult Analyte
Grace van der Gugten
Provincial Health Services Authority
Long Abstract | Bio | View Video
For the most part, LC-MS/MS assay development and validation is, and should be, a within-laboratory project. However, the new user can find assay development a daunting task. We will describe the LC method development for one of our more challenging to measure endogenous steroids: serum/plasma aldosterone. We will discuss selection of mobile phases, column selection, gradient program development, interference testing, and addition of cortisol to the method. Importantly, we will cover unexpected workflow challenges and continual improvements and discoveries we have made while the assay has been in clinical production.
3:50 PM
4:15 PM
Targeted Quantitative Mass Spectrometric Immunoassay for Analysis of Serum Amyloid A (SAA) in Human Plasma
Olgica Trenchevska
Arizona State University
Long Abstract | Bio | View Video
Proteins can exist as multiple proteoforms in vivo that can have important roles in physiological and pathological states. Presented here is the development and characterization of mass spectrometric immunoassay for quantitative determination of serum amyloid A (SAA) proteoforms. Intra- and inter-day precision and recovery characteristics of the assay were established, yielding CVs<10%. The new assay was benchmarked against existing SAA ELISA, producing 2.2% Altman-Bland bias. We used the assay to determine the individual concentrations of the SAA proteoforms across a cohort of ~ 300 samples, revealing 7 different SAA genetic polymorphic types and a total of 18 different proteoforms.		Development, Implementation, and Stress Testing of a Rapid HPLC-HRAMS Screening Method for Detection of Twenty Antiretroviral (ARV) Compounds in Human Serum
Autumn Breaud
The Johns Hopkins University
Long Abstract | Bio | Financial Disclosure | View Video
The aim of this work was to develop a method for cost-effective and rapid screening for the presence of a panel of ARV drugs using a multiplexed HPLC-HRAMS approach. Validation studies were performed before preparation and analysis of multiple, large batches of study samples. Upon increased throughput of the method, we found that a number of components of the assay required better life cycle definition. We also found a number of challenges associated with the acquisition mode of the instrument when panel components share fragments and when there may be an isotopic peak interference from one precursor mass to another.		Lipidomics of Eicosanoids Highlights Infection and Inflammation Progression
Edward A. Dennis
University of California, San Diego
Long Abstract | Bio
The largest numbers of distinct molecular species in cellular metabolism are the lipids where tens of thousands of distinct molecular species exist in cells/tissues. We have developed novel liquid chromatographic-mass spectrometric based lipidomics techniques termed “CLASS” to solve lipidomics problems, often in the context of an overall omics analysis of immunologically-activated macrophages integrating transcriptomics, proteomics, and metabolomics of lipid metabolites. Our laboratory has developed a robust and comprehensive approach to the lipidomics analysis of hundreds of fatty acids, acylethanolamines and inflammatory eicosanoids. We have built on our previous application of lipidomic analysis to characterize “synergistic” cellular lipid signaling of Toll-like (TLR) and purinergic receptors in stimulated macrophages as models of bacterial infection and inflammation.		PANEL: Diagnostic Gaps in Infectious Diseases: A Proposal for an Interdisciplinary Approach to Develop Mass Spectrometry Methods to Find the Bug and Treat the Host
Andy Hoofnagle Et Al. (see Long Abstract)
University of Washington
Long Abstract
The application of Mass Spectrometry in the clinical microbiology laboratory has been limited mainly to the identification of microorganisms growing in culture. However, there are a large number of technological gaps in both the diagnosis and management of infections that could potentially make ideal candidates for MS-based solutions. In this panel discussion, three clinical microbiologists will present clinical problems and/or unmet diagnostic needs in the field of clinical microbiology. A panel of experts in MS will then respond to each need presented, weighing in on the potential feasibility of a MS-based analytical solution. The major objective of this session is to foster interdisciplinary collaborations to improve both the diagnosis and management of infections using novel MS-based assays.		Part 2b: Serum Aldosterone: An LC Method Development Case Study for a Difficult Analyte
Grace van der Gugten
Provincial Health Services Authority
Long Abstract | Bio | View Video
For the most part, LC-MS/MS assay development and validation is, and should be, a within-laboratory project. However, the new user can find assay development a daunting task. We will describe the LC method development for one of our more challenging to measure endogenous steroids: serum/plasma aldosterone. We will discuss selection of mobile phases, column selection, gradient program development, interference testing, and addition of cortisol to the method. Importantly, we will cover unexpected workflow challenges and continual improvements and discoveries we have made while the assay has been in clinical production.
4:15 PM
7:00 PM
RECEPTION
@ Exhibit Hall
Enjoy mingling with colleagues and Exhibitors.

Take time to explore the Posters.

Heavy Appetizers and Drinks to be provided.
Sponsored by:
Thermo

5:00 PM
6:00 PM
POSTERS
@ Exhibit Hall
Odd-Numbered Posters to be Attended & Presented.

All Posters should be removed by 7:30 PM.
7:00 PM
8:00 PM

CORPORATE WORKSHOPS (Evening)
Thermo Scientific
Harbor Ballroom 2

Break the Bottleneck - Accelerate Genomics to Proteomics to Bedside with New Cloud-computing and Informatics Solutions
Mazi Mohiuddin, Senior Applications Scientist Thermo Fisher Scientific, Biomarker Research Initiatives in MS (BRIMS)

Pre-Register

An overview of the proteo-genomic data analysis workflows and automation of informatics solutions for clinical service providers as well as researchers to run routine data analysis workflows in a seamless manner.

Agilent Technologies
Harbor Ballroom 3

The Agilent StreamSelect LC/MS System – Up To Four Times the Throughput with Outstanding Reliability
Agilent Technologies

Pre-Register

The StreamSelect LC/MS System delivers up to four parallel chromatographic separations to the same triple quadrupole mass spectrometer, with superior robustness and data quality. Intuitive automation software coordinates the completely integrated system, maximizing MS utilization and greatly enhancing throughput and cost-effectiveness.

IONICS MS
Marina 6

Recent Developments in Endocrinology – Diurnal Steroids Fluctuations, Free 25-OH Vitamin D3 & Thyroid Hormone Management
Dr. Steven Soldin

Pre-Register

In this workshop Dr. Steven Soldin from the National Institutes of Health (NIH) will address several recent developments in endocrinology. Topics discussed will include: diurnal fluctuations of steroids, changing trends in management of thyroid patients, the role in measurement of free 25-OH Vitamin D3 and a variety of clinical applications.
8:00 PM
10:00 PM
HOSPITALITY
@ Shoreline Patio by the Pool
Enjoy the San Diego evening down by the Marina with heaters and fire pits.
Drinks provided.
10:00 PMTUESDAY CLOSED

WEDNESDAY
5:45 AM
6:45 AM
YOGA
@ Nautilus 5
Energize yourself for the day! Yoga is a complimentary offering for all MSACL registrants.
A limited number of yoga mats will be provided.
6:00 AM
8:15 AM
BREAKFAST
@ Harbor Island Foyer
Enjoy a light continental breakfast that tastes even better after morning Yoga or a run along the bay, or maybe it tastes just fine by itself.
7:00 AM
8:00 AM

CORPORATE WORKSHOPS (AM)
Spark Holland
Seabreeze

(1) Routine Clinical LC-MS Assays Using Online Sample Preparation: Daily User Experiences and Illustrations
(2) Fully Automated DBS and DPS Analysis by LC/MS
Martijn van Faassen, UMCG, The Netherlands (1). Jack Henion, Rob Sturm and Regina Oliveira, Quintiles BioAnalytical and ADME Labs, Ithaca, NY 14850 (2).

(1) For reproducible highly sensitive LC-MS/MS assays, sample preparation is pivotal. Several sample prep approaches are available, each with its own (dis)advantages. In our clinical chemistry laboratory different sample prep strategies are used with online sample prep as chosen method. In this presentation we provide an overview of our clinical LC-MS/MS assays using online sample prep and discuss different strategies by focusing on relevant examples. (2) Punching of DBS cards for LC/MS analysis is tedious and too manual for a high sample volume lab. This presentation describes how commercial DBS cards as well as a prototype dried plasma spot (DPS) card can be analyzed on-line in a fully automated fashion using either SRM LC/MS or high res. mass spectrometry techniques for the bioanalytical determination of drugs in micro blood samples.

Sigma-Aldrich
Spinnaker

Developing the Best Sample Preparation Methods to Ensure Robust LC/MS Analyses in Biological Fluids
Craig R. Aurand, Supelco/Sigma-Aldrich

Sample preparation continues to be one of the critical factors for effective method development when analyzing biological samples. Too often this portion of the assay is not allocated sufficient attention to ensure a robust analytical method. The goal of this seminar is to discuss several approaches for sample preparation for biological fluids, and to demonstrate the benefit that proper sample clean up can have on an LC/MS based methods. Sample preparation methods for matrices such as plasma and urine will be covered, along with techniques such as solid phase extraction, phospholipid depletion, and enzymatic digestion.
GENERAL SCIENTIFIC SESSION 5
Track 1
Harbor Ballroom 1
Proteomics: Protein Variants
Chair: John Mills		Track 2
Harbor Ballroom 2
Small Molecules: Sample Prep
Chair: Brian Rappold		Track 3
Harbor Ballroom 3
Metabolomics 5: Inborn Errors of Metabolism
Chair: Elizabeth Payne		Track 4
Marina 6
Sample Prep: Proteomic Workflow
Chair: Mark Duncan		Track 5
Executive Center
Basics: Proteomics
Chair: TBA
8:30 AM
8:55 AM
Identification of Insulin Like Growth Factor 1 (IGF1) Variants Using High-Resolution Accurate-Mass Mass Spectrometry (HRAM-MS)
Hemamalini Ketha

Mayo Clinic
Long Abstract | Bio | View Video
We demonstrate how high-resolution-accurate-mass mass spectrometry (HRAM-MS) identifies IGF1 variants (V-IGF1) that immunoassays identify as wild type IGF1. IGF1 measured with immunoassay-measured-IGF1 (IGF1-IA) in patients with V-IGF1 are higher than with IGF1-MS. In 15 out of 2480 (0.6%) patients half of IGF1-IA was identified as V-IGF1. Of note, one patient with V-IGF1 was receiving recombinant growth hormone (rGH) for treatment of Noonan Syndrome with a protein tyrosine phosphatase 11 (PTPN11) mutation. Identification and accurate quantitation of V-IGF1 is of clinical value especially for patients receiving GH therapy, those exhibiting GH resistance or IGF1 mutations. This presentation will describe the identification, characterization of V-IG1.		One SLE to Measure Them All: Two-part Elution for Analysis of Urinary HIAA, HVA and VMA
Zlatuse Clark
ARUP Laboratories
Long Abstract | View Video
The monoamine acids 5’-hydroxyindoleacetic (HIAA), homovanillic (HVA), and vanillylmandelic (VMA) are metabolites of serotonin and the catecholamine neurotransmitters. Laboratory measurement of these acids in urine is used to assess overproduction of the parent amines due to neuroendocrine tumors. We developed and validated a method for HIAA, HVA, and VMA using SLE-based sample preparation and LC-MS/MS analysis. Development of the sample preparation method required significant diversion from manufacturer guidelines, but resulted in the use of a single product to extract all analytes. Sequential two-part elution of the analytes yielded cleaner extracts and higher recoveries. The final method is robust and fast. 		Preliminary Study on Clinical Application of Metabolomics for Laboratory Diagnosis of Inborn Errors of Metabolism
Qin Sun
Baylor College of Medicine
Long Abstract | Bio | Financial Disclosure | View Video
To meet the challenge of integrating metabolomic test in diagnostic labs, we developed a rapid metabolomic workflow to analyze plasma from patients with a confirmed inborn error of metabolism (IEM). Analysis was performed using a non-targeted multi-mass spectrometry platform. The analytes detected encompass a number of classes of important small molecule biomarkers such as fatty acids, acylcarnitines, amino acids, bile acids, carbohydrates, lipids and nucleotides, etc. Metabolic profiling was able to correctly diagnose 20 of the 21 disorders. In an additional set of prior unresolved cases, metabolomic analysis detected disturbances that pointed to a genetic disorder or assisted in the interpretation of concurrent DNA analysis. In summary we have demonstrated the clinical utility in the diagnosis of IEMs as well as functional confirmation of genetic and genomic results of uncertain significance.		High-throughput and Reproducible Workflows to Prepare Human Plasma Samples for Proteomic Analysis
Jennifer van Eyk
Advanced Clinical Biosystems Institute, Cedars Sinai
Long Abstract | Bio
We have implemented an automated peptide preparation protocol on a liquid handling workstation (Biomek NXP ) coupled with an SRM workflow using triple quadruple mass spectrometer (QTRAP® 6500 system). The complete analysis had a coefficient of variation of less than 10% based on assessment of the internal beta-galactosidase standard. We present automated high-throughput workflows with sample processing and enrichment robotics for accurate and reproducible large-scale analysis of biological/clinical samples. This workflow can be applied to biomarker validation, drug response monitoring, disease state and progress monitoring.		Part 1: Introduction to Clinical Proteomics
Timothy Collier
Cleveland HeartLab, Inc.
| View Video
This session is directed toward attendees new to mass spectrometry and its use for the measurement of proteins and peptides (proteomics) in the clinical laboratory. It will be presented in three parts with questions and discussion welcomed after each. Part I of this session will focus on the definition of proteomics within the broader context of systems biology. We will also discuss how information gathered from proteomic measurements can offer clinical insight into a patient’s state of health, compared and contrasted to genetic, metabolite, and other small molecule diagnostics.
8:55 AM
9:20 AM
Mass Spectrometry of Hemoglobin Variants
Jane Yang
UCSD
Long Abstract | Financial Disclosure
Hemoglobin variants caused by a single nucleotide polymorphism are mutant forms of hemoglobin that typically result in a single amino acid substitution, such as hemoglobin S. Two classic techniques used in the clinical laboratory, cation exchange (CE)-HPLC and capillary zone electrophoresis (CZE), are oftentimes insufficient to identify specific variants. In addition to the CE-HPLC and CZE results, we use both top-down and bottom-up mass spectrometry (MS) approaches to confirm the identities of hemoglobin variants at the intact subunit and peptide levels, by using mass differences and fragmentation spectra to predict the amino acid substitution and location, respectively.		Elimination of Matrix Effects Using Mixed-mode SPE Plate for High Throughput Analysis of Free Arachidonic Acid in Plasma by LC-MS/MS
Warren Chen
Bonna-Agela Technologies
Long Abstract | View Video
The matrix effects on the analysis of arachidonic acid in plasma on LC-MS/MS was investigated by comparing various sample pretreatment methods including protein precipitation, LLE, single-mode SPE, and mixed-mode SPE. The results indicated the last one with Cleanert MAS-M gave better results in terms of eliminating matrix effect of phospholipids and proteins in plasma. The optimized method was applied on real human plasma. The study demonstrated that a mixed-mode SPE plate could be adapted to high throughput sample pretreatment of hydrophobic analytes in plasma which are usually co-eluted with phospholipids and proteins on reversed phase HPLC column.		High-Performance Chemical Isotope Labeling LC-MS for Clinical Metabolomics
Liang Li
University of Alberta
Long Abstract | Bio | View Video
In clinical metabolomics, both targeted and untargeted analyses are being used for monitoring the metabolic changes associated with a disease. However, there are still several technical challenges in untargeted metabolome profiling. In particular, it is difficult to detect, quantify and identify a large number of metabolites that are present in a metabolomic sample with a wide range of concentrations and diverse physicochemical properties. In this presentation, a high-performance chemical isotope labeling liquid chromatography mass spectrometric platform for quantitative and comprehensive metabolome profiling of biological systems will be described. Some selected applications of this platform for clinical metabolomics studies including disease biomarker discovery will be presented.		Automated High-throughput Clinical Proteomics Workflow Using Hydrogel Nanotrap Particle Technology and Mass Spectrometry Analysis
Matthew Rosenow
Translational Genomics Research Institute
Long Abstract | Financial Disclosure | View Video
The functionality of a high-throughput automated preparation workflow that incorporates a novel nanoparticle technology is presented. Serum samples were spiked with cytokines and other small cell signaling proteins, and processed using an automated system. Levels of the protein spikes used to generate response curves ranged from the pg/mL to ng/mL levels and were within the physiological relevant levels of the proteins. SRM mass spectrometry analysis using corresponding isotopically labeled peptides, show low total process CV’s at the various levels, and show the physiological relevant concentration of the proteins to be within the linear region of the response curves. This study demonstrates a simple clinical proteomics workflow that does not require antibody-based target enrichment or complex sample processing that is also capable of producing clinical grade process CVs. 		Part 2: Introduction to Clinical Proteomics
Timothy Collier
Cleveland HeartLab, Inc.
| View Video
A mass spectrometer is more than a box with specimens going in and a numbers coming out. This part of the session will discuss a few of the most used types of mass spectrometers in the clinical laboratory and how they perform their measurements in the context of proteomic diagnostics. This includes: (1) Protein/Peptide ionization, (2) Guiding ions through the instrument, (3)Fragmentation, (4) Detection, and (5) Databases and Data Interpretation Tools
9:20 AM
9:45 AM
Analysis of Hemoglobin Variants by Top-down Mass Spectrometry
Didia Coelho Graça

University of Geneva
Long Abstract | Bio
A high-resolution top-down mass spectrometry method was developed for the detection of mutated hemoglobin chains using selected diagnostic product ions for data interpretation. This procedure brings more precise information about the considered hemoglobin variants (proteoforms) than any current protein analysis methods used for hemoglobin disorders diagnosis. The method was successfully applied to the analysis of hemoglobin β chain variants carrying various single point mutations and an Aγ-β fusion protein. The results showed that the developed data analysis process allows fast and reliable interpretation of top-down electron transfer dissociation mass spectrometry data by non-expert users in the clinical area.		Ultra-rapid, Fully-automated Plasma Clozapine and Norclozapine Analysis Using AC Extraction Plate Technology and Flow-injection MS/MS
Lewis Couchman

King's College Hospital
Long Abstract | Bio | Financial Disclosure | View Video
Therapeutic drug monitoring (TDM) of plasma clozapine and N-desmethylclozapine (norclozapine) is well-established. Fully-automated sample preparation using novel AC Extraction PlatesTM and automated liquid handling (both Tecan Schweiz AG) combined with flow-injection MS/MS of extracts minimises analysis times. The data capture time was 5 seconds per sample. Results were comparable to those produced using manual extraction and LC-MS/MS. Use of deuterated internal standards compensated for matrix effects for both analytes. The principle embodied in this approach may have wide application.		Metabolic Profiling of Developing Infants: Term vs. Preterm at Birth
Frances Jackson

Imperial College London
Long Abstract | Bio
Advances in neonatal medicine have seen an increase in the survival of babies born preterm. This has resulted in a significant burden to health and education services with increased risk for several childhood and later life diseases. To understand these conditions, urine and stool samples from infants born term (> 37 weeks gestation) or preterm (24 - 36 weeks gestation) at birth have been analysed using UPLC-MS techniques. Samples were taken from birth until up to 3 months of age giving us the metabolic profiles of infant’s development. This will provide insight and identify biomarkers which may be prognostic of developmental conditions including adverse health outcomes that manifest later in life. 		Nanoporous Substrates-Enabled, Functional Mechanism-Based Method for the Early Diagnosis in Cancer Diseases
Tony Hu

Houston Methodist Research Institute
Long Abstract | Bio | View Video
Circulating peptides have been recognized as useful signatures that can be traced to cancer-specific metabolic or post-translational modification events at early-stage tumor progression. We have established “Nanotrap” to effectively fractionate blood peptides with little to no sample processing. By coupling this technique to advanced mass spectrometry, we can bypass the limitation of current proteomic technologies, by “amplifying” the amount of small peptides extracted from blood samples. Our cutting-edge nanotechnologies coupled with advanced mass spectrometry and customized biostatistical analysis facilitated the functional mechanism-driven peptide biomarker studies for revealing the early events associated with the signature mutations or pathways in tumor progression.		Part 3: Introduction to Clinical Proteomics
Timothy Collier
Cleveland HeartLab, Inc.
| View Video
The final part of the session will feature examples from the literature of diagnostics employing mass spectrometry based proteomics, highlighting unmet clinical needs addressed in the development of such assays. The session will conclude with an overview of new techniques and technologies that present opportunities for further advancement in the field.
9:45 AM
10:45 AM
COFFEE BREAK
@ Harbor Island Foyer
Take a break and get a coffee, juice, water and/or small snack.

Commune with colleagues or perhaps go for a short walk outside by the water to refresh for the next session.
GENERAL SCIENTIFIC SESSION 6
Track 1
Harbor Ballroom 1
Proteomics: Clinical Assays II
Chair: Andy Hoofnagle		Track 2
Harbor Ballroom 2
Analysis of Chemical/Drug Exposure
Chair: Mark Marzinke		Track 3
Harbor Ballroom 3
Metabolomics 6: Clinical Metabolomics
Chair: Kevin Cho		Track 4
Marina 6
Organism Identification
Chair: Susan Butler-Wu		Track 5
Executive Center
Metabolomics for Newbies
Chair: Caroline Johnson
10:45 AM
11:10 AM
Trials and Triumphs in the Development of a Quantitative Assay for Amyloid-beta Peptides
Mari Demarco

St Paul's Hospital & Univ. British Columbia
Long Abstract | Bio
While Alzheimer’s disease has a defined pathology on autopsy, in vivo diagnosis is challenging—particularly in early stages of disease when treatment opportunities are greatest. Toward the development of an in vivo diagnostic model, cerebrospinal fluid biomarkers amyloid-beta and tau proteins have been extensively studied and are now included in research diagnostic criteria. Historically, quantitation of amyloid-beta peptides has relied on immunometric techniques. Herein we describe our antibody-free LC-MS/MS workflow to quantitate amyloid-beta peptides in cerebrospinal fluid. We will also present a method comparison between LC-MS/MS and a commonly used commercial ELISA, as well as results from a case-control study.		Steroidomic Footprinting Based on UHPLC Qualitative and Quantitative High-resolution MS for the Evaluation of Endocrine Disrupting Chemicals in H295R Cells
David Tonoli

University of Geneva
Long Abstract | Bio | View Video
UHPLC coupled to high-resolution MS approaches were devised for the simultaneous untargeted screening and quantification of a selected subset of steroids in H295R cell culture supernatant exposed to different concentrations of a potential endocrine disrupting chemical, triclocarban (TCC). Chemically-driven feature selection with database matching followed by multivariate analysis led to a selection of the most important steroids perturbed by TCC. This approach indicates that TCC affects an early step in steroid biosynthesis. This strategy was devised to be compatible with high-throughput screening and stratification of potential endocrine disruptors.		A Bioinformatic Package for Automated Targeted and Global Profiling Analysis of Large Direct Infusion Mass Spectrometry Datasets
Gonçalo Correia

Imperial College London
Long Abstract | Bio
Direct infusion mass spectrometry based methods (DIMS) are being increasingly deployed for high-throughput and cost effective analysis of large sample sets. However, computational tools for automated data pre-processing and analysis tailored for this particular type data are almost nonexistent. We present a new open-source package written in the Python programming language for the automated pre-processing and analysis of DIMS data. It contains modules for pre-processing, targeted quantification and untargeted/profiling analysis. Its development was prompted by the ongoing DIMS analysis of more than 10,000 24 hour urine collection samples from the INTERMAP study.		Improved Turnaround Time for Identification of Blood Culture Isolates Using MALDI-TOF MS to Assay "Scum", or Brief Outgrowths of Blood Culture Broths
Mark Gonzalez

Washington University in St. Louis
Long Abstract | Bio | View Video
Prompt and appropriate treatment for bloodstream infections results in improved patient outcomes; this is facilitated by early identification of the causative pathogen. MALDI-TOF MS offers an accurate and rapid method for microbial organism identification. To further expedite identification of bloodstream pathogens, MALDI-TOF was performed on early subculture growth, or “scum” growth, from positive blood cultures, prior to the formation of isolated colonies. We found that this easy, rapid, and cost effective method resulted in a significant reduction in time for microorganism identification for yeast, non-fermenting Gram-negative bacteria and Enterobactericeae isolated from blood culture specimens.		Metabolomics for Newbies
Caroline Johnson & Julijana Ivanisevic

The Scripps Research Institute
Long Abstract | View Video
Part 1: In this course we will introduce mass spectrometry-based metabolomics. We will cover the importance of metabolomics in science research, the diversity of the metabolome, and challenges of global metabolomics. Mass spectrometry technologies will be covered and include electrospray ionization mass spectrometry coupled to liquid chromatography which is making a significant impact in metabolomics. We will cover untargeted and targeted approaches, data processing, including profile alignment, statistical analysis and metabolite identification. To follow on from the basics of metabolomics, the biomedical applications of metabolomics will be discussed for biomarker discovery, pharmacometabolomics and integration of other –omic technologies with metabolomics. This course is designed to introduce the non-expert to metabolomics and its potential applications of the field.
11:10 AM
11:35 AM
A Mass Spectrometric Immunoassay Coupled to Selected Reaction Monitoring Reveals Novel Relationships Between Plasma PCSK9 and Metabolic Phenotypes in Patients
Benoit Coulombe
Institut de recherches cliniques de Montréal-IRCM
Long Abstract | View Video
Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a key regulator of blood low density lipoprotein cholesterol (LDL-C) levels and circulates in various forms that affect PCSK9's function and thereby LDL-C levels. Commercial PCSK9 ELISA assays do not allow discrimination between the various forms of the protein. We have developed and applied a robust and sensitive Mass Spectrometric ImmunoAssay coupled to Selected Reaction Monitoring assay for the absolute quantification of all PCSK9 domains and a posttranslational modification in plasma of two distinct human cohorts. This assay revealed novel relationships between PCSK9 and metabolic phenotypes, as compared to classical ELISA assays. 		Meconium Fatty Acid Ethyl Ester, Ethyl Glucuronide, and Ethyl Sulfate Sensitivity and Specificity to Detect Maternal Drinking During Pregnancy
Sarah Himes
Quest Diagnostics Nichols Institute
Long Abstract | Bio | View Video
Meconium fatty acid ethyl esters (FAEE), ethyl glucuronide (EtG), and ethyl sulfate (EtS) were quantified by liquid chromatography-tandem mass spectrometry in the same meconium sample (n=107). Moderate-substantial agreement between maternal self-reported drinking after 19 weeks gestation and meconium EtG >30 ng/g was observed (kappa: 0.57, 95% CI 0.41-0.73). This marker and associated cutoff was superior to a 7 FAEE sum >2 nmol/g and all other individual and combination marker cutoffs. With meconium EtG >30 ng/g as the standard and maternal self-report beyond 19 weeks gestation as the test condition, 82% sensitivity and 75% specificity were observed. These data indicate maternal alcohol consumption in the second half of pregnancy is best represented by meconium EtG >30 ng/g. We recommend meconium EtG as a better alcohol marker than FAEE for identifying prenatal alcohol exposure.		Metabolomics and Mitochondria in Human Diabetic Nephropathy
Kumar Sharma
University of California San Diego and the Veteran
| Bio | Financial Disclosure | View Video
We have taken a urinary-based targeted metabolomic approach to identify biomarkers for diabetic nephropathy. We identified that a panel of metabolites that are characteristic of patients with diabetes and reduced renal function. These metabolites comprise a large network of interacting pathways and largely indicate a reduction in mitochondrial function. Several novel therapeutic agents provide renoprotection and are able to stimulate mitochondrial biogenesis and increase urinary levels of the metabolomic panel of diabetic kidney disease. These studies suggest that metabolomics is a powerful platform to identify new therapeutic targets and to monitor drug development for diabetic complications. 		Identification of Nocardia Species by MALDI-TOF Mass Spectrometry
Brian Bird

ARUP Laboratories
Long Abstract | Bio | View Video
MALDI-TOF mass spectrometry for identification of Nocardia species remains challenging. However, routine sample preparation on young cultures using an up-to-date commercial database minimally augmented with custom spectra allowed 82% (64 of 78), 89% (8 of 9), and 94% identification to species, complex, and genus level, respectively. This indicates that special sample preparation and custom databases may be unnecessary for routine Nocardia spp. identification. 		Metabolomics for Newbies
Caroline Johnson & Julijana Ivanisevic

The Scripps Research Institute
Long Abstract | View Video
Part 2: In this course we will introduce mass spectrometry-based metabolomics. We will cover the importance of metabolomics in science research, the diversity of the metabolome, and challenges of global metabolomics. Mass spectrometry technologies will be covered and include electrospray ionization mass spectrometry coupled to liquid chromatography which is making a significant impact in metabolomics. We will cover untargeted and targeted approaches, data processing, including profile alignment, statistical analysis and metabolite identification. To follow on from the basics of metabolomics, the biomedical applications of metabolomics will be discussed for biomarker discovery, pharmacometabolomics and integration of other –omic technologies with metabolomics. This course is designed to introduce the non-expert to metabolomics and its potential applications of the field.
11:35 AM
12:00 PM
Development of a Reference Measurement System for Urine Albumin
Ashley Beasley Green
National Institute of Standards and Technology
Long Abstract | Bio
Urinary excretion of albumin is a major diagnostic and prognostic marker of renal dysfunction and cardiovascular disease; therefore, accurate measurement of urine albumin is vital to clinical diagnosis. To address urine albumin measurement precision, we have developed the following components of the urine albumin reference measurement system: a multiplexed candidate reference measurement procedure that utilizes isotope dilution-mass spectrometry (ID-MS) and multiple reaction monitoring (MRM) to quantify urine albumin; a primary reference material to be used as a calibrator for higher-order urine albumin methods; and a secondary reference material to be used as a matrix-based quality control for commercially-available urine albumin assays. 		The Utility of High Resolution Mass Spectrometry in Identifying Potential Chemical Culprits of Synthetic Cannabinoid Epidemics
Roy Gerona
University of California, San Francisco
Long Abstract | View Video
We report the use of LC-QTOF/MS in the rapid identification of novel synthetic cannabinoids associated with three epidemics in the last three years. Effective collaboration between an analytical toxicology lab, Poison Centers, DPH, DEA and the CDC along with Cayman Chemical, a reference standard provider, in one series of cases allowed the effective use of both non-targeted and targeted screening workflows to identify the novel drug common to patient samples that is associated with the toxidrome observed.		Pre-diagnostic Biomarkers and Clinical Biochemistry of Lung Cancer
William Wikoff
UC Davis Genome Center
Long Abstract | Bio
The potential for metabolomics to yield blood-based biomarkers relevant to lung cancer screening and early detection has not been previously investigated. We applied an untargeted metabolomics approach to identify pre-diagnostic biomarkers using sera from a large patient cohort in a blinded discovery mode from current or former heavy smokers, resulting in a single, highly significant potential biomarker. Blinded validation was performed using an independent set of samples and matched controls. Tissue level changes were investigated by comparing normal to matched biopsy tissue from the same patient. Metabolomics revealed key perturbations in multiple pathways associated with early stage lung adenocarcinoma.		Evaluation of Sample Preparation Methods, Instrumentation, and Databases for the Identification of Rapid Growing Mycobacterium spp... Using MALDI-TOF MS
Allison McMullen

Washinton University in St. Louis
Long Abstract | Bio | View Video
We evaluated different sample preparation methods, instrumentation platforms (VITEK MS and Bruker Biotyper), and databases for MALDI-TOF MS identification of 64 rapid growing Mycobacterium spp. (RGM) cultivated on solid media. Bruker Biotyper correctly identified 62/64 isolates. No identification was obtained for two isolates. The VITEK MS provided no result for one isolate and one incorrect identification (Saramis database) and no results for four isolates (v3.0 database). The VITEK-MS sample preparation required less hands-on time and had high confidence values/scores for the RGM isolates tested. MALDI-TOF MS will be able to expedite the identification of RGM from clinical specimens. 		Metabolomics for Newbies
Caroline Johnson & Julijana Ivanisevic

The Scripps Research Institute
Long Abstract
Part 3: In this course we will introduce mass spectrometry-based metabolomics. We will cover the importance of metabolomics in science research, the diversity of the metabolome, and challenges of global metabolomics. Mass spectrometry technologies will be covered and include electrospray ionization mass spectrometry coupled to liquid chromatography which is making a significant impact in metabolomics. We will cover untargeted and targeted approaches, data processing, including profile alignment, statistical analysis and metabolite identification. To follow on from the basics of metabolomics, the biomedical applications of metabolomics will be discussed for biomarker discovery, pharmacometabolomics and integration of other –omic technologies with metabolomics. This course is designed to introduce the non-expert to metabolomics and its potential applications of the field.
12:00 PM
1:00 PM
LUNCH
@ Harbor Island Foyer
Lunch to be provided in the Harbor & Bayview Foyers.

• Get ready to join a Corporate Workshop at 1:00 PM.
1:00 PM
2:00 PM

CORPORATE WORKSHOPS (PM)
Shimadzu
Harbor Ballroom 1

Prepare Your Laboratory for the Future - Laboratory-on-a-Card Technology and Ultra-Fast Mass Spectrometry
Fred Regnier (Novilytic Labs), Scott Kuzdzal (Shimadzu Scientific Instruments)

Please join us for this interactive workshop where you will discover a new technology (Noviplex Duo) that enables multiple plasma extractions and sample preparation steps to be performed from a single blood drop in just minutes. This “lab-on-a-card” technology simplifies and accelerates sample preparation by: - Integrating and automating multiple sample prep steps, - Introducing in-transit sample preparation, - Allowing collection of multiple fractions, - Reducing sample prep time, - Simplifying sample transport, - Reducing chain of custody issues, and - Enabling simultaneous analysis of proteins and metabolites. Combined with ultra-fast mass spectrometry, these technologies can save your laboratory time, money and resources. Attendees will receive a lunch, t-shirt and GiantMicrobe!

Thermo Scientific
Harbor Ballroom 2

Employing Translational Research Workflow on LC-HRAM Platform for Detection of Pathogen Induced Cancer in a Human T-Cell Leukemia Virus Type 1 Disease Model
Sucharita M. Dutta, Ph.D. – LeRoy T. Canoles Jr. Cancer Research Center, Eastern Virginia Medical School

Pre-Register

Dr. Sucharita Dutta, from Eastern Virginia Medical School will discuss the Human T-Cell Leukemia Virus Type 1 (HTLV-1) as the causative factor for the development of an aggressive lymphoma, Adult T-cell Leukemia (ATL). The translational workflow involves standard data-dependent acquisition (DDA) experiments followed by more in-depth pSMART data acquisition methodologies or the most comprehensive global profiling of proteins from exosome samples to exhaustively mine for proteins that show functional significance via pathway analysis.

SCIEX
Harbor Ballroom 3

1) Lipid Clinical Biomarkers: From Discovery to Quantitation. 2) Implementing LC-TOF-MS/MS in the Clinical Research Arena Developing A Broad Spectrum Test
1) Paul RS Baker, PhD, SCIEX. 2) Kara Lynch, PhD, UCSF

1) Lipid Clinical Biomarkers: From Discovery to Quantitation - This presentation will focus on the general methods used for lipid biomarker discovery will be presented with special focus on the novel use of DMS as a tool orthogonal to chromatography to achieve better qualitative lipid identification. 2) Implementing LC-TOF-MS/MS in the clinical research arena developing a broad spectrum test - This presentation will focus on the pros and cons of assorted screening approaches for maximum compound coverage, minimizing false positive finding and improving confidence in results. As well as, tips for managing challenging cases that include complex samples and matrices.

2:00 PM
2:30 PM
COFFEE BREAK
@ Harbor Island Foyer
Take a break and get a coffee, juice, water and/or small snack.

Commune with colleagues or perhaps go for a short walk outside by the water to refresh for the next session.
GENERAL SCIENTIFIC SESSION 7
Track 1
Harbor Ballroom 1
Proteomics: Dried Blood Spots
Chair: Irene van den Broek		Track 2
Harbor Ballroom 2
Small Molecule Analysis 3
Chair: Jason Sawyer		Track 3
Harbor Ballroom 3
Metabolic Phenotypes of Organ Dysfunction
Chair: Nathaniel Mahieu		Track 4
Marina 6
New Advances in Ionization
Chair: Jane Yang		Track 5
Executive Center
Clinical Microbiology & MALDI-TOF
Chair: Carey-Ann Burnham
2:30 PM
2:55 PM
The Development of a Proteomic Wellness Assay Using Dried Blood Spots: Moving Clinical Protein Diagnostics Towards Personalized Reference Ranges
James Bollinger

University of Washington - Genome Sciences
Long Abstract | Bio | View Video
Dried blood spot (DBS) sampling represent and attractive technique suitable to meet the demands of both personalized medicine and widespread epidemiological research. We describe method development parameters for the multiplexed analysis of a panel of clinically relevant proteins in DBS samples via selected reaction monitoring. For each protein target, we derive a set of optimal peptides for tandem MS analysis and demonstrate the value of our approach by designing and validating an SRM assay for the multiplexed quantitation of a set of 50 proteins within DBS samples using a single protein as an internal standard.		A Novel Method for Plasma Metanephrine Analysis Using the Waters Unispray Ionisation Technique
Joanne Adaway
University Hospital South Manchester
Long Abstract | Bio | Financial Disclosure | View Video
Measurement of plasma metanephrines is useful in the diagnosis of paragangliomas, but many assays require a large volume of plasma due to poor assay sensitivity, and often require lengthy sample preparation. We developed a method using the Waters Online Solid Phase Extraction manager coupled to a Waters Xevo TQS with a Unispray source that required only 50 µL of sample. Validation was carried out according to FDA guidelines and found to be acceptable, with an LLOQ of 18.75 pmol/L for metanephrine and 20.2 pmol/L for normetanephrine. We believe that this method is suitable for use in a busy clinical laboratory.		Metabolic Reprogramming and Lipid Distribution Protects Gclm KO Mice from Alcohol Induced Steatosis
Srujana GOLLA
National Cancer Institute, NIH
Long Abstract | Bio | View Video
The present study elucidated the protective role of antioxidant glutathione against chronic alcohol induced steatosis using the integrated analysis of metabolomics and lipidomics. The biochemical adaptation and lipid distribution patterns of wild-type and gamma-glutamylcysteine synthetase (Gclm)-null mice on chronic alcohol exposure were analyzed by robust UPLC coupled with ESI-QTOF mass spectrometry. The data matrix generated was further analyzed by multivariate data analysis using SIMCA 13+ software for identifying differential makers underlying the disease. The depleted glutathione was to shown impair de novo fatty acid and alters lipid distribution patterns in the Gclm-null mice upon alcohol exposure. 		Clinical Applications of Top-down Proteomics
Daojing Wang
Newomics Inc.
Long Abstract | Bio | Financial Disclosure
The penetration of MS-based proteomics, particularly top-down proteomics, into the in vitro diagnostics market has remained low. MS-based platform has to achieve the: 1) sensitivity, 2) throughput; and 3) robustness, comparable to or even better than those of ELISA in order to find wider clinical acceptance. We have developed the silicon-based monolithic multinozzle emitter array (MEA) chip for high-sensitivity and high-throughput nanoflow-LC-ESI/MS. In this talk, I will present new MEA chip-enabled workflows and assays, for direct top-down proteomics analysis of sub-microliter volumes of human blood samples, and demonstrate the utilities of our assays in diagnosis and monitoring of several diseases. 		Part 1: Cases in Clinical Microbiology and MALDI-TOF Mass Spectrometry
Allison Mcmullen1 And Lori Bourassa2
1Washington University School of Medicine, St. Louis MO and 2University of Washington, Seattle WA
| Bio | View Video
Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is quickly becoming the primary method of microorganism identification in many clinical laboratories. It is rapid, accurate, and cost-effective. In addition, the increased species-level resolution made possible by using this method, especially for Gram-positive bacilli and infrequently encountered taxa, is helping to inform clinicians and laboratorians about the biology and clinical significance of many bacterial species. In this session, we will present {interactive} case studies that demonstrate both the diagnostic pitfalls and the clinical utility of MALDI-TOF MS and "interesting" results acquired as a result of widespread use of this identification method.
2:55 PM
3:20 PM
In Pursuit of ‘Normal Baselines’: Longitudinal Measurement of Protein Biomarkers in Dried Blood Spots
N. Leigh ANDERSON
SISCAPA Assay Technologies
Long Abstract | Bio | Financial Disclosure | View Video
Precise, longitudinal measurement of protein biomarkers in dried-blood-spots (DBS) is an attractive option for providing preventative, personalized medicine at reasonable cost. Here we present an automated SISCAPA workflow for multiplexed measurement of 20 proteins in longitudinal DBS samples collected from 10 individuals. By reducing the total workflow CV for all proteins below their ‘normal’ biological coefficient of variation, we were able to define the ‘normal’ range for each analyte in the longitudinal DBS samples. The data suggests that each individual possesses a unique ‘protein-fingerprint’ that if monitored longitudinally can provide invaluable insight into the person’s state of wellness and/or disease. 		Recurrent Need for a Robust Method for MeasuringT3/rT3 by LC-MS/MS: An Exercise in Madness or a New Beginning for a Misused Marker?
Julie Ray
ARUP Laboratories
Long Abstract | Bio | View Video
Even though measurement of reverse T3 (rT3) remains controversial in the diagnosis of hypothyroidism, the test continues to attract attention for managing an underactive thyroid in the integrative medical community. We present an evaluation of our sample preparation method for measuring T3/rT3 by LC-MS/MS and the frequent fouling of the mass spectrometer associated with it. Removal of pigmentation in 30-40% samples (suspected from bilirubin), with the use of 1% formic acid in dichloromethane helped maintain cleanliness of the instrument. A new application for the measurement of rT3 and T3 was tested in the CSF (cerebro spinal fluid) of brain injury patients. Preliminary results indicated that measurement of rT3 may have utility in brain injury in addition to the diagnosis of hypothyroidism.		Metabolomics Study of Premature Labor: Combined Supercritical Fluid Chromatography and Liquid Chromatography Coupled with Ion Mobility-Mass Spectrometry
Rafael Montenegro-Burke

Vanderbilt University
Long Abstract | Bio
The cause of preterm birth is not entirely understood, and several factors (maternal age, infections, etc.) can influence the gestation period. Therefore, discovering a set of delivery date predictive biomarkers would improve the diagnosis of preterm risk pregnancies. In order to address this, an unbiased, untargeted metabolomics study of both term and preterm amniotic fluid samples was performed. The combination of liquid chromatography (LC) and supercritical fluid chromatography (SFC) coupled to ion mobility-mass spectrometry (IM-MS) demonstrates the great advantage of higher metabolome coverage, which increases the probabilities of detecting significant differences between sample groups in complex biological samples.		Rapid Quantitative Biomedical Testing with Ambient Mass Spectrometry
Jentaie Shiea
National Sun Yat-sen University
Long Abstract | Bio | View Video
Ambient mass spectrometry is a technique that operates the ionization source under ambient conditions to characterize analytes with minimum or no sample pretreatment. However, applying AMS for accurately quantitative analysis is still a problem, since the sample quantity is inconsistent for each analysis. In this study, an AMS technique - thermal desorption-electrospray ionization/mass spectrometry combined with solid-phase microextraction was developed for quantifying trace drugs in biological fluids. SPME/TD-ESI/MS contains features from both SPME and TD-ESI/MS, including simple and fast extraction, concentration, and characterization. Since the quantity of analytes extracted by SPME is dependent on the surface area of the SPME fiber, a combination of SPME with AMS makes it possible to perform rapidly quantitative analysis for certain types of drugs and chemical compounds in biofluids.		Part 2: Cases in Clinical Microbiology and MALDI-TOF Mass Spectrometry
Allison Mcmullen1 And Lori Bourassa2
1Washington University School of Medicine, St. Louis MO and 2University of Washington, Seattle WA
| View Video
Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is quickly becoming the primary method of microorganism identification in many clinical laboratories. It is rapid, accurate, and cost-effective. In addition, the increased species-level resolution made possible by using this method, especially for Gram-positive bacilli and infrequently encountered taxa, is helping to inform clinicians and laboratorians about the biology and clinical significance of many bacterial species. In this session, we will present {interactive} case studies that demonstrate both the diagnostic pitfalls and the clinical utility of MALDI-TOF MS and "interesting" results acquired as a result of widespread use of this identification method.
3:20 PM
3:45 PM
Systematic Comparison of Internal Standard Platforms for Absolute Protein Quantification of Cytokines by MRM-MS
Kerry Scott

NIST
Long Abstract | Bio
Protein quantification methods based on the principle of multiple reaction monitoring can provide absolute protein quantification values. These methodologies are broadly applicable to clinical biomarkers, systems biology, and the pharmaceutical industry; which require accurate and precise protein abundance measurements. An evaluation of three conventional internal standards (AQUA peptides, QconCAT constructs, and PSAQ proteins) for quantitative accuracy, precision, and inherent advantages and limitations was conducted. A strategy for methodological improvement to current internal standard platforms was also investigated. The results of these studies as well as take home lessons will be presented.		Performance of Symmetric Dimethylarginine Against Other Markers of Kidney Function
Joe El-Khoury

Yale University
Long Abstract | Bio | View Video
Glomerular filtration rate (GFR) is the best overall index of kidney function. Knowledge of GFR is essential to the diagnosis, classification, and management of kidney disease. Serum measurements of cystatin C (cysC), creatinine and their equations are the most widely used indirect markers, while symmetric dimethylarginine (SDMA) is an emerging marker. The objective of this study was to compare the performance of SDMA with cysC, creatinine and their equations to direct GFR measurement by radioactive iothalamate. A total of 40 subjects were included in this study. A published LC-MS/MS method was used for measuring SDMA. CysC and creatinine were measured by Roche Cobas 8000 (Indianapolis, IN). CysC showed the best overall correlation with GFR (r=0.92) and highest AUC (0.923) for kidney donor eligibility, followed by SDMA (r=0.87, AUC=0.882) then creatinine (r=0.76, AUC=0.767).		In vivo Global Isotope Metabolomics Implicates the Arginase Pathway in Ischemic Retinopathy
Caroline Johnson

The Scripps Research Institute
Long Abstract | Bio
Proliferative diabetic retinopathy (PDR) is the most severe form of diabetic retinopathy, here, global isotope metabolomic analysis was used to investigate metabolism to understand the ocular metabolic landscape. Analysis of vitreous humor from patients revealed an upregulation of arginine metabolism in PDR patients compared to non-diabetic controls. The oxygen-induced-retinopathy (OIR) mouse model revealed similar dysregulation.Validation by targeted metabolomics and the analysis of a second set of patient samples confirmed the upregulation of the same metabolites. The metabolic fate of U-15N-arginine determined in the OIR model, revealed a predominance of the arginase pathway to form proline. These results indicate that in PDR, the arginase pathway predominates, decreasing the availability of arginine for NO synthesis a metabolite required for adequate endothelial cell function. 		Challenges in Rapid Evaporative Ionization of Breast Tissue: A Novel Method for Real-time MS Guided Margin Control During Breast Surgery
Julia Balog

Imperial College London
Long Abstract | Bio | Financial Disclosure | View Video
A novel setup based on Rapid Evaporative Ionisation Mass Spectrometry has been developed for the analysis of heterogeneous breast tissue. This technique is suitable with almost 100% accuracy for the separation of normal breast tissue, benign alterations (fibroadenoma) and different breast cancers. The novel setup enables both the use of cutting and coagulation electrosurgical settings, allowing us to record and analyze REIMS data throughout the whole breast surgery. The technique can be used for MS guided breast surgery, likely reducing the number of positive margins and therefore decreasing local regional cancer recurrence and necessary re-operations.		Part 3: Cases in Clinical Microbiology and MALDI-TOF Mass Spectrometry
Allison Mcmullen1 And Lori Bourassa2
1Washington University School of Medicine, St. Louis MO and 2University of Washington, Seattle WA
| View Video
Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is quickly becoming the primary method of microorganism identification in many clinical laboratories. It is rapid, accurate, and cost-effective. In addition, the increased species-level resolution made possible by using this method, especially for Gram-positive bacilli and infrequently encountered taxa, is helping to inform clinicians and laboratorians about the biology and clinical significance of many bacterial species. In this session, we will present {interactive} case studies that demonstrate both the diagnostic pitfalls and the clinical utility of MALDI-TOF MS and "interesting" results acquired as a result of widespread use of this identification method.
3:45 PM
7:00 PM
HOSPITALITY
@ Shoreline Patio by the Pool
Enjoy the San Diego evening down by the Marina with heaters and fire pits.

Dinner and Drinks provided. Some of the best wine you may ever taste will also be provided as a sweetener for staying the whole conference through.
7:00 PMCONFERENCE CLOSED