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MSACL 2014: Preliminary Conference Program

Sheraton Hotel & Marina • San Diego, CA • March 1-5, 2014

With Thanks to Our Corporate Sponsors:
Thermo Bruker Agilent Waters   AB SCIEX SimulTOF Shimadzu

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. Space is limited.
6:30 AM
8:00 AM
BREAKFAST
@ Harbor Island Foyer
8:15 AM
10:00 AM

SHORT COURSES: SESSION 1

Introduction to Clinical Mass Spectrometry
Robert Kobelski, PhD
Level: 1 (Beginner)
Location: Executive 2
General Toxicology
Jeffery H. Moran, PhD
Level: 1 (Beginner)
Location: Executive 4
** All day Saturday. Ends at noon on SUN.
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: Nautilus 1-2
How to Develop Robust Assays Faster Using Free Data Analysis Tools
Fred Lytle, PhD
Level: 2 (Intermediate)
Location: Spinnaker
A Comprehensive Review of Clinical Mass Spectrometry Technology & Techniques, including Miniaturization
Jack Henion, PhD
Level: 2/3 (Intermediate/Advanced)
Location: Executive 3
Targeted Proteomics
Andy Hoofnagle, MD, PhD, Mike MacCoss, PhD & Nathan Yates, PhD
Level: 3 (Advanced)
Location: Harbor Ballroom 1
** Starts at 1:00 PM on SAT
Development and Validation of Quantitative LC-MS/MS Assays for Use in Clinical Diagnostics
Russell Grant, PhD & Brian Rappold
Level: 3 (Advanced)
Location: Harbor Ballroom 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
11:30 AM

SHORT COURSES: SESSION 2

11:30 AM
1:00 PM
LUNCH
@ Harbor's Edge Restaurant

Group A at 11:30 AM (All except below).
Group B at 12:15 PM (Grant, Hoofnagle, Lytle).
45 minute lunch
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
8:30 PM
PRIVATE EVENT:Travel Awardee Reception & Dinner
@ Grande Ballroom Foyer

** This event is exclusively for Travel Awardees. **
Dinner and Reception from 6:00 - 8:30 PM.

Thermo Agilent Waters Bruker
8:30 PM
10:00 PM
HOSPITALITY
@ Grande Ballroom Foyer

A bit rainy tonite so we will be inside at the Grand Ballroom Foyer. We go back outside to the Shoreline patio tonite, weather permitting.
8:30 PM
10:00 PM
YOGA & MEDITATION
@ Nautilus 5

Yoga is a complimentary offering for all MSACL registrants.
Participants welcome at anytime during the class.
A limited number of yoga mats will be 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. Space is limited.
6:30 AM
8:00 AM
BREAKFAST
@ Harbor Island Foyer
8:15 AM
10:00 AM

SHORT COURSES: SESSION 1

Preparing Manuscripts for Publication: Improving Your Chances for Success
Thomas Annesley, PhD
Level: 0 (General Interest)
Location: Executive 1
How to Process Body Fluids for LC-MS/MS Analysis of Small Molecules
Karl-Siegfried Boos, PhD
Level: 2 (Intermediate)
Location: Marina 6
Metabolomics
Gary Siuzdak, PhD & Gary Patti, PhD
Level: 3 (Advanced)
Location: Seabreeze
Introduction to Clinical Mass Spectrometry
Continued from Saturday
Robert Kobelski, PhD
Level: 1 (Beginner)
Location: Executive 2
General Toxicology
Continued from Saturday
Jeffery H. Moran, PhD
Level: 1 (Beginner)
Location: Executive 4
** All day Saturday. Ends at noon on SUN.
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: Nautilus 1-2
How to Develop Robust Assays Faster Using Free Data Analysis Tools
Continued from Saturday
Fred Lytle, PhD
Level: 2 (Intermediate)
Location: Spinnaker
A Comprehensive Review of Clinical Mass Spectrometry Technology & Techniques, including Miniaturization
Continued from Saturday
Jack Henion, PhD
Level: 2/3 (Intermediate/Advanced)
Location: Executive 3
Targeted Proteomics
Continued from Saturday
Andy Hoofnagle, MD, PhD, Mike MacCoss, PhD & Nathan Yates, PhD
Level: 3 (Advanced)
Location: Harbor Ballroom 1
** Starts at 1:00 PM on SAT
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 Ballroom 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 PM
1:00 PM
LUNCH
@ Harbor's Edge Restaurant
12:00 PM
1:00 PM
PRIVATE EVENT: Travel Awardee Lunch
@ Shoreline Patio

** This event is exclusively for Travel Awardees. **
Lunch from 12:00 - 1:00 PM.
1-2 min Welcome Introductions from each Travel Award sponsor at 12:15 PM.

Thermo Agilent Waters Bruker
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.

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 4
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, which will be attended from 5:00 - 6:00 PM. Heavy Appetizers and Drinks to be provided.

POSTER PRESENTERS: If you are presenting a poster your poster should have been up at 4:15 PM
Sponsored by:
Thermo
5:00 PM
6:00 PM
POSTERS
@ Exhibit Hall

ALL Posters Attended from 5:00 - 6:00 PM.
PLENARY LECTURE SERIES
@ Harbor Ballroom
Chair: Anthony Craig
7:00 PM
8:00 PM
Commercialization and Protection of Intellectual Assets in Applications of Mass Spectrometry
Ben Borson
Borson Law Group, PC
Long Abstract | Biography
Bringing improvements in assessing conditions using Mass Spectrometry (MS) requires substantial investment in research, development of devices and methods for diagnostic and therapeutic interventions. Investments are unlikely to be made unless returns on those investments can be realized. Returns are made possible through licensing of patented inventions, trade secrets, and other intellectual assets (IP). With proper strategies combining business, technology, and the law, commercializing important innovations will result in improved patient care, recognition of innovators' contributions, and business success, all of which can lead to further innovation and improvements in health care.
8:30 PM
10:00 PM
HOSPITALITY
@ Shoreline Patio

Enjoy the San Diego evening down by the Marina with heaters and fire pits. Drinks provided. At Shoreline Patio by the pool.
8:30 PM
10:00 PM
YOGA & MEDITATION
@ Nautilus 5

Yoga is a complimentary offering for all MSACL registrants.
Participants welcome at anytime during the class.
A limited number of yoga mats will be 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. Space is limited.
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

Poster presenters for Monday must have their posters placed by the end of breakfast (8 AM).
7:00 AM
8:00 AM

CORPORATE WORKSHOPS (AM)

IsoSciences
Marina 6

Deficiencies of Deuterium as an Isotopic Label in MS Standards
Scott Landvatter, Ph.D.

Deuterium is the most frequently used isotope in stable isotope labeled standards. It is generally easy to incorporate and is typically less expensive than synthesizing standards with other stable isotopes such as 13C and/or 15N. However, deuterium suffers from inherent drawbacks that can limit the accuracy or viability in quantification by LC/MS/MS. Among these problems are loss or exchange of deuterium under chemical or MS conditions, ‘messy’ molecular ions and LC or GC co-elution problems. A related issue surrounds the confusion of what exactly is meant by “percent isotope incorporation.” This workshop will focus on these issues and discuss strategies for overcoming these problems by selecting appropriate label location, synthesis methodology and choice of isotope.

Tecan
Seabreeze

Automated Sample Preparation for Mass Spectrometry
Guy Burssens, Robert Wohleb and Roland Geyer

Although there have been great advances in mass spectrometry (MS) instrumentation in recent years, its unglamorous counterpart, sample preparation, has not enjoyed the same rate of development. Sample preparation has become a major bottleneck, and the issues associated with manual processing have hindered the uptake of MS innovation in the life science industry. To help overcome this, we will show how the Tecan Freedom EVO®-based end-to-end process automation for even the most challenging protocols will liberate you from the bottleneck of manual sample preparation. We will also discuss the newly released Tecan AC Extraction Plate with TICE™ (Tecan Immobilized Coating Extraction) technology which will revolutionize your sample preparation routine for LC-MS analysis of small molecules, and present you applications examples.

DPX Labs
Spinnaker

Rapid Sample Preparation using Dispersive Pipette Extraction (DPX) with LC/MS/MS Analyses for Clinical Applications
Dani C. Mata, Orange County Crime Lab, Santa Ana, CA; Dr. Oscar G. Cabrices, Gerstel, Inc., Linthicum, MD; Dr. William E. Brewer, DPX Labs, LLC Columbia, SC

The DPX workshop will highlight more efficient methods for sample preparation by discussing three applications using DPX tips with LC/MS/MS analyses. 1. Analysis of 22 Benzodiazepines and 3 ‘Z-Drugs’ in 5 Matrices using DPX WAX Tips. 2. Automated DPX Extractions of Pain Management Drugs in Urine and Oral Fluid. 3. Ultrafast Sample Preparation and Analysis of 25-OH-Vitamin D2 and D3 in Serum.

8:10 AMWELCOME, INTRODUCTION & ORIENTATION
@ Harbor Ballroom

Welcome, Introduction and Orientation

Time to get this show on the road!

Updates on two MSACL conference apps (beta stage) that will hopefully make your contact collecting and Scientific Program browsing much easier ... now and in the future.

Updates on new developments on the direction MSACL is headed.

And finally, the introduction of the Recognition Award Committee.

PLENARY LECTURE SERIES
@ Harbor Ballroom
Chair: Gary Siuzdak
8:30 AM
9:15 AM
Direct Mass Spectrometric Characterization of Fluids, Cells and Tissues - the Benefits and the Price of Real-Time Analysis
Zoltan Takats
Imperial College London
No Long Abstract
Development of ambient mass spectrometric techniques opened a new way of looking at clinical samples regarding the simplicity (and time demand) of analysis and the nature of data produced by these techniques. Application of these approaches in cancer surgery, histopathology, clinical microbiology and clinical chemistry has been successfully demonstrated. While these techniques mean valid alternatives to currently used technologies - with clear benefit on both cost and reliability sides - their widespread application can potentially change the current landscape of medical diagnostics. Future role of these methods in stratified clinical patient journeys will be discussed with particular emphasis on the currently unresolved problems and their future solutions regarding both regulatory and technology aspects.
9:15 AM
10:00 AM
When Clinical Lab Tests Fail: New Paradigms of Metabolic Regulation
Gary Patti
Washington University School of Medicine
No Long Abstract
Traditionally, mass spectrometry-based metabolomics has been applied to identify metabolites whose levels are altered as a function of disease. While this platform has provided great insight into many pathologies, it fails to identify important changes in metabolites whose levels are homeostatically controlled by multiple converging pathways. Here we describe an isotope-based platform that enables tracking of pathway dynamics at the systems level. To accomplish this approach, we have developed an extension of the widely used XCMS metabolomic software that processes data from isotopically enriched samples in an automated fashion. We show that monitoring isotope incorporation at the systems level provides new insights into cancer pathogenesis that would otherwise be missed by standard metabolomic approaches.
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
Protein Analysis by Mass Spectrometry
Chair: Leigh Anderson
Track 2
Harbor Ballroom 2
High Throughput Analysis of Low Abundance Steroids
Chair: Christoph Seger
Track 3
Harbor Ballroom 3
Microbiology I
Chair: Nathan Ledeboer
Track 4
Marina 6
Basics: Setting Up a Clinical Lab
Chair: Rob Fitzgerald
10:45 AM
11:10 AM
Automated Mass Spectrometry Analysis of Full-length Hemoglobin for Clinical Applications
Alexander Scherl
Geneva University Hospitals
Long Abstract | Bio | Financial Disclosure
We developed a method for the identification of hemoglobin variants by electron-transfer dissociation (ETD) of the full length protein (Coelho-Graça et al, JASMS, 2012) and a method for the quantification of Hemoglobin A2 using precursor-ion isolation and detection (Acosta-Martin et al., Anal. Chem., 2013). These two workflows were now integrated into a fully automated pipeline, which includes data acquisition for HbA, HbS, HbC identification and HbA2 relative quantification. This workflow will now be tested in a clinical environment for hemoglobin variant identification as well as diagnosis of thalassemia.
High-Throughput Sensitive LC-MS/MS Estradiol Analysis
Laura Owen
University Hospital of South Manchester
Long Abstract | Bio
We have developed a LC-MS/MS assay for the measurement of estradiol and estrone. The assay has a novel sample preparation utilising on-line solid phase extraction combined with a short chromatography column that results in a rapid run time making it suitable for routine clinical use. Despite the rapid run time, estradiol and estrone are adequately separated to remove a potential isobaric interference. The simultaneous measurement of estradiol and estrone gives additional clinical information towards calculating total estrogen status.
Development of an Assay and Viral Database for the Identification of Biothreat Viruses Using MALDI Mass Spectrometry of Intact Virions
Lisa Cazares
USAMRIID
Long Abstract | Bio
Since many laboratories employ the MALDI Biotyper system for bacterial identification, we sought to develop a viral typing assay which would be compatible with this platform. We employed a strategy of producing spectra from highly purified viral stocks for spectral database compilation, which could then be used to type unknowns using a more rapid method of harvest. A custom made set of reference spectra was successfully generated from 7 highly purified viral isolates using a modified BioTyper method. Spectral patterns were unique to each virus and peaks corresponding to the molecular weight of capsid proteins were detected and confirmed by LC/MS-MS. Successful identification of West Nile virus, and Rift Valley Fever virus was achieved from 1ml of infected media using a sucrose cushion and standard micro-centrifugation with BioTyper scores of 1.8 and above.
So You Want a Mass Spectrometer?
Deborah French
University of California San Francisco
Long Abstract | Bio
Use of liquid chromatography-mass spectrometry for analysis of small molecules in the clinical laboratory has increased in the past few years. Many labs have considered implementation of this technology, but some do not have enough experience to decide in which type of mass spectrometry system they should invest. This presentation will describe the basics of liquid chromatography, the types of mass spectrometer available and what type of analysis they are most suited to (quantitative versus qualitative), and the basics of return on investment calculations that may be required to justify a mass spectrometer purchase to laboratory administration.
11:10 AM
11:35 AM
Quantitation of Monoclonal Antibody Therapeutic Infliximab by LC-MS/MS
Maria Alice Willrich
Mayo Clinic
Long Abstract | Bio
We have verified that unique proteotypic peptides from variable regions of Infliximab (chimeric IgG1 kappa targeting TNF-α) may be used to quantify the drug in patient serum using standard SRM analysis on an ABSciex API 5000. The assay was developed, validated and compared to an electrochemiluminescent method. Samples from patients receiving Infliximab therapy were tested at various time-points following treatment. The ability to quantify Infliximab in patients using proteotypic peptides and LC-MS/MS was demonstrated. This analytical approach has the potential to be quickly adaptable to other monoclonal antibody drugs and to significantly improve patient care.
Three Years Experience in Screening and Diagnosis of Primary Aldosteronism by LC-MS/MS
Daniel Holmes
St. Paul's Hospital
Long Abstract | Bio | Financial Disclosure
Primary aldosteronism (PA) is the most common form of secondary (ie potentially treatable) hypertension and is generally caused by adrenal hyperplasia or aldosterone producing adenoma. Biochemical testing for this disease has traditionally relied on immunoassays for aldosterone and plasma renin activity or renin mass. However these have been fraught with problems of intermethod variation and analytical cross-reactivity. LC-MS/MS offers an excellent solution. We describe three years experience in screening, diagnosis and tumor localization for primary aldosteronism by LC-MS/MS. We delineate the technical challenges associated with the methods, data processing, and autointerpretation. Advantages and potential pitfalls will be described along with the the many fortuitous diagnoses provided by LC-MS/MS which would have otherwise been delayed or missed.
Salvage Microbiology: Direct Detection of Pathogens from Sterile Clinical Specimens Submitted for Culture Following Initiation of Antimicrobial Treatment
John Farrell
University of Illinois College of Medicine
Long Abstract | Bio
We prospectively identified 56 cases of suspected bacterial infection from which 80 sterile specimens were submitted for culture after the initiation of antimicrobial treatment (multiple specimens were submitted for culture in 21 cases). Compared to culture, PCR/ESI-MS was more likely to detect potential pathogens: 74% (59/80) vs. 32.5% (26/80). Pathogenic organisms were detected by PCR/ESI-MS that were not identified in culture in 45% (25/56) of the patients included in our study: 36% (20/56) of the PCR/ESI-MS positive patients had completely negative cultures; and 9% (5/56) had polymicrobic infections. Cohen's Kappa statistic confirms that there is discordance between culture and PCR/ESI-MS results (K = 0.348). Our prospective series demonstrates that PCR/ESI-MS can detect pathogenic bacteria that are not grown in conventional culture directly from clinical specimens obtained following initiation of antibiotic treatment.
Instrument Selection, Due Diligence, and Setting Up a Clinical Lab for Successful Implementation of MS
Robert Fitzgerald
UCSD
Long Abstract | Bio
After determining that your laboratory is ready to expand into clinical applications of mass spectrometry and determining what type mass spectrometer will be optimal for the work you propose, the next tasks are to select a vendor, make necessary modifications to your lab, perform instrument qualification, and begin validating clinical assays. This presentation will focus on our laboratories approach to performing due diligence and to setting up a laboratory to successfully implement clinical applications of mass spectrometry.
11:35 AM
12:00 PM
Identification and Quantitation of Insulin Analogues by Immunocapture Coupled with LC-MS/MS
Grace van der Gugten
St. Paul's Hospital
Long Abstract | Bio
Insulin is frequently measured for the investigation of spontaneous hypoglycaemia in adults and children. Several synthetic insulin analogues are in routine clinical use for the management of Type I and Type II diabetes mellitus. In clinical medicine, hypoglycemia remains one of the most frequently encountered sequelae of insulin therapy. Commercial insulin immunoassays exhibit variable cross-reactivities to analogue insulins. Given these limitations, we have developed an immuocapture-liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the identification and quantification of five insulin analogues. We will present our work in developing this assay, and its application to clinical and forensic cases in our laboratory.
Aldosterone by LC-MS/MS: Cost Saving Not at the Cost of Patient Care
Julie Ray
ARUP Laboratories
Long Abstract | Bio
Measurement of low abundance endogenous analytes in high throughput laboratories present challenges associated with specificity, efficiency of analysis and cost of testing. We evaluated Liquid Liquid Extraction (LLE) vs Supported Liquid Extraction (SLE) in measuring serum aldosterone using an AB SCIEX Triple Quad 5500 Mass Spectrometer with and without the use of differential ion mobility spectrometry (DMS). Under the evaluated conditions, over 30% of samples analyzed by various methods had ratio of the mass transitions outside of the expected limits which lead us to evaluate several HPLC column chemistries and mobile phases. LLE and SLE extractions produced comparable results. LLE was 30% more laborious although less expensive. The use of LLE and DMS could reduce expenses but at the cost of high starting serum volume. Our final choice for the assay using low sample volume and low cost was in favor of LLE in conjunction with 2 dimensional chromatography and use of an AB Sciex 5500 MS without the use of DMS.
Computational Considerations for Intensity-Based Microbial Identification Using MALDI-TOF MS
Chris Crutchfield
The Johns Hopkins University School of Medicine
Long Abstract | Bio
MALDI-TOF driven microbial identification is rapidly changing the workflow of clinical microbiology laboratories. The majority of users will understand that the problem the analyzer is attempting to solve is rooted in pattern recognition. The complexities of how this is achieved computationally may be more problematic to understand intuitively. This presentation will approach this computational problem using the historically well performing dot product approach of spectral library search algorithms. The caveats explored will be evaluating how performance characteristics change due to m/z window size, baseline subtraction algorithms, and comparison of median-based and eigenvector based reference spectrum generation. These permutations were evaluated based on the bootstrapped comparison of spectra of E. avium and E. Faecium generated by a Bruker Biotyper.
The MS Test Life Cycle Process: How to Keep Your Lab Moving Forward
Paul Jannetto
Mayo Clinic
Long Abstract | Bio
As more laboratories develop mass spectrometry-based tests, they need to have a standardized program to define, document, validate, and monitor laboratory-developed tests going forward to prepare for and meet the increased regulatory oversight. This presentation will describe the seven phases of a mass spectrometry test life cycle process at the Mayo Clinic which takes a laboratory-developed test from design, development, verification, validation, launch, and maintenance to retirement. It will also focus on the importance of post analytical monitoring and how proficiency testing programs can be used in combination with other assessments to monitor assay performance over time.
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.
• Poster Group 'a' to present during the 2:00 - 3:00 PM coffee break in Exhibit Hall.
1:00 PM
2:00 PM

CORPORATE WORKSHOPS (PM)

Agilent Technologies
Harbor Ballroom 1

Ultrafast SPE-QTOF Methods for Analyzing Panels of Drugs in Urine and Leading Proteomics into the Age of Automation: High-throughput Protein Sample Preparation for Mass Spectrometry Applications Enabled by the AssayMAP Bravo Automation Platform
Dr. Vaughn Miller, Agilent Technologies and Dr. Jason Russell, Agilent Technologies

In this workshop 2 speakers will present SPE and Automated Sample Preparation and how these approaches increase throughput for LCMS Analyses. Dr. Vaughn Miller will discuss Ultrafast analytical methods for drug panels in urine with as many as 35 analytes that have been successfully developed using a SPE-QTOF system using RapidFire High-Throughput MS System. Dr. Jason Russell will discuss the challenges with protein/peptide sample preparation for LCMS analysis. To address these issues routine sample handling tasks common to LC/MS proteomic workflows have been automated using the Agilent AssayMAP Bravo platform and its suite of proteomic tools engineered to bring reproducibility, scalability, protocol portability, and ease-of-use to LC/MS sample preparation.

Thermo Scientific
Harbor Ballroom 2

Transformational Advances in Clinical Research Mass Spectrometry - Novel Approaches to Quantitation with New HPLC and Orbitrap MS Technology
Dr. Roy Peake, Boston Children's Hospital; Dr. William Clarke, Johns Hopkins Medical Center; Dr. David C. Kasper, Medical University of Vienna

Pre-Register

In this workshop three speakers will describe how they utilized the latest LC/MS technologies to unravel common challenges faced by clinical research and forensic toxicology labs. Dr. Roy Peake will discuss a multisite study designed to determine the effect of standardized LC/MS protocols on variability of results obtained in analysis of immunosuppressant drugs. Dr. William Clarke will describe a single injection method to positively identify and quantify triclosan, parabens and PFCs using high resolution accurate mass (HRAM) MS instrumentation. Dr. David Kasper will discuss TurboFlow technology, including staggered injection and high-resolution accurate-mass LC-MS for high-throughput analysis compounds.

Waters
Harbor Ballroom 3

Improving Steroid Hormone Measurements: Addressing the Challenge with LC/MS
1. The Importance of Standardization of Hormone Measurements
Hubert Vesper, CDC

2. Multiplexed Analysis of Steroid Hormones in Human Serum using Novel Microflow Tile Technology and LC-MS/MS
Jessica Prenni, Colorado State University

3. Routine Steroid Hormone Analysis with Online Sample Preparation
Brian Keevil, University Hospital of South Manchester

For decades, laboratory physicians and research scientists have recognized that conventional antibody-based immunoassays have limitations when measuring sex steroid hormones, especially when used to assess new treatments or to address new research questions. Today, LC/MS based solutions are being deployed by clinical researchers for routine sex steroid hormone measurements. Properly implemented, routine LC/MS assays overcome the recognized limitations of immunoassays and may exceed other performance criteria. Join us for this informative workshop to learn about the history of the Endocrine Society’s and CDC’s path toward excellence in steroid hormone analysis and how two laboratories have implemented Waters technologies to develop tests for steroid hormone analysis.

Shimadzu
Marina 6

Prepare Your Laboratory for the Future - Next-Generation Plasma Collection Technology, Mass-Linked Immuno-Selective Analysis and Ultra-Fast Mass Spectrometry.
Fred Regnier - Professor Emeritus & John H. Law Distinguished Professor of Chemistry Purdue University and Novilytic Laboratories, Rachel Lieberman - Senior Product Specialist, Shimadzu Scientific Instruments and Scott Kuzdzal - Life Science Business Manager, Shimadzu Scientific Instruments

This workshop will provide your organization with facts and critical lessons needed for the successful adoption and integration of “lab-on-a-card” plasma collection technology, ultra-fast & automated protein digestions and ultra-fast mass spectrometry in your laboratory. You will discover a new Noviplex Card technology (Novilytic Labs) that enables rapid plasma generation in just minutes without the need for venipuncture, centrifugations, etc. We will also introduce Perfinity Workstation and integrated Digestion Platform technologies that enable automated, reproducible protein digests on a sub-minute timescale, greatly improving the applicability of peptide based SRM assays. These technologies, used with ultra-fast mass spectrometry save your laboratory time, money and resources and will greatly improve data reproducibility and quality.

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

Group 'a' Posters to be attended from 2:00 - 3:00 PM.
GENERAL SCIENTIFIC SESSION 2
Track 1
Harbor Ballroom 1
Proteins: MS to the Clinical Lab - Why and How
Chair: Bob Bergen
Track 2
Harbor Ballroom 2
Designer Drugs: Getting Your Mass Spectrometer High (Resolution)
Chair: Kara Lynch
Track 3
Harbor Ballroom 3
Metabolomics I: Applications to Cancer
Chair: Gary Siuzdak
Track 4
Marina 6
Basics: Standardization & Harmonization
Chair: Juli Botelho
3:00 PM
3:25 PM
Comparing LC/MS and Immunoassays for Clinical Biomarkers
Michael Lassman
Merck & Co
Long Abstract | Bio
In clinical studies, biomarker assays are routinely measured using immunoassays. These assays generally meet the criteria necessary for clinical biomarkers, but in some cases are not sufficient. Mass spectrometry based assays are inherently more selective and while these assays generally lack sensitivity compared to immunoassays, the selectivity of the platform has advantages in the ability to multiplex, and independently measure PTMs and isotope labeled tracers as part of kinetics studies. Recent technical advances such as immuno-affinity enrichment and low-flow chromatography allow mass spectrometry to compete with immunoassays in terms of sensitivity.
Moving High Resolution Mass Spectrometry into Routine Clinical Practice: Optimizing Targeted Data Analysis
Katie Thoren
University of California, San Francisco
Long Abstract
Although there has been much interest in high resolution mass spectrometry (HRMS), a major challenge in implementing this technology in the clinical laboratory is the data analysis process. Even for targeted analysis, the process is complex because so much data is generated from a single sample and many parameters are involved in identifying compounds. Using a training set and a test set of urine samples, we try to determine the optimal parameters for targeted data analysis. The goal of this study is to streamline the data analysis process and make HRMS more accessible to the clinical lab.
Metabolomics and Metabolite Imaging Correlates Biofilm Polyamine Synthesis with Colon Cancer
Caroline Johnson
The Scripps Research Institute
Long Abstract | Bio
Mass spectrometry-based metabolomics was used to investigate the effects of colonic biofilms on metabolites in colon cancer. Untargeted analysis revealed upregulation of polyamines in tumors compared to normal tissues, which was enhanced in biofilm positive tumors. Antibiotic-treated tumors were metabolically similar to biofilm negative than positive tumors, indicating that polyamines were produced by both host and biofilm. Targeted analysis obtained concentrations of these metabolites, and nanostructure initiator mass spectrometry (NIMS) imaging provided spatial specificity of the metabolites in the tissues. The results show that biofilms can alter the colon metabolome to produce known regulators of cell proliferation and tumor growth.
CDC Efforts Towards the Development of a Reference Method for Measurement of 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 in Serum
Ekaterina Mineva
Centers for Disease Control and Prevention
Long Abstract
Vitamin D status is routinely assessed by measuring serum concentrations of the most stable and abundant vitamin D metabolites. To assure the accuracy of measurements and to harmonize laboratory results regardless of measurement procedure or location, worldwide standardization of 25-hydroxyvitamin D measurements is underway. Several JCTLM-recognized reference measurement systems have been established consisting of reference materials and methods. We are presenting key details from our new isotope dilution LC-MS/MS candidate reference method for measurement of 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 in serum. The method will be used to support standardization activities, including the CDC Vitamin D Standardization Certification Program.
3:25 PM
3:50 PM
Next Generation Blood Plasma Collection and Sample Preparation in Mass Spectrometry Based Diagnostics
Fred Regnier
Novilytic Laboratories
Long Abstract | Bio
A new technology is described that allows blood plasma extraction from a single 25-50 uL drop of blood obtained from a finger-stick. Blood fractionation was achieved by capillary driven flow through multiple membrane layers that sequester blood cells via filtration as plasma migrates to a collection disc at the bottom of the membrane stack. Fractionation is completed in roughly 3 min when a 6.4 mm diameter collection disc is saturated with either a 2.4 or 4.8 uL sample of plasma, depending on the disc size. Analyte concentration and sampling volumes are hematocrit independent. Stripping (delaminating) the upper, cell bearing membrane layers allowed evaporation from the plasma loaded collection disc to dryness in 15 min. Internal standards and antibodies were either preloaded in the collection disc or added after plasma sampling; allowing affinity selection and analysis of analytes with a Perfinity Workstation (PWS) coupled to a Shimadzu 8050 triple quadrapole instrument.
Determination of Metabolites of the Bath Salt Pentylone, Using LC/MSAll
Jessica Lloyd
Washington University
Long Abstract | Bio
Bath salts have emerged recently as recreational drugs, which are known to cause hallucinogenic effects. The pathways metabolizing bath salts have not been well characterized, which complicates our ability to screen for bath salt use. We sought to understand the metabolism of the bath salt pentylone by using a mouse model and a novel untargeted metabolomic platform based on LC/MSAll. MSAll alternates between high and low energy scans during acquisition, resulting in MS1 data and fragmentation data for each precursor ion. We used MS1 data to quantify metabolite alterations in pentylone-treated mice and MSAll data to obtain semi-putative structural assignments.
Colorectal Cancer Detection Using Targeted Metabolic Profiling
Jiangjiang Zhu
University of Washington
Long Abstract | Bio
Colorectal cancer (CRC) is one of the most prevalent and deadly types of cancer worldwide. In this study, we proposed a targeted LC-MS/MS metabolic profiling approach for sensitive and selective CRC diagnosis. 234 serum samples from three groups (66 CRCs, 76 polyps, and 92 controls) were analyzed. 42, 48 and 8 out of 158 targeted metabolites showed statistical significance between pairwise group comparisons. PLS-DA models were established for the separation of CRC patients from other groups, and validated using Monte Carlo cross validation. ROC curves were generated, with excellent AUROCs (>0.93), high sensitivities (>0.89) and high specificities (>0.80).
An Update on Standardization Efforts for Steroid Hormones
Hubert Vesper
CDC
Long Abstract
Correct diagnosis and treatment of patients rely on accurate testing. Furthermore, the effective use of research findings in patient care and public health requires testing in research and patient care to be accurate and comparable. The CDC, together with the Partnership for the Accurate Testing of Hormones (PATH), is working to improve the accuracy and reliability of steroid hormone testing. In addition to its testosterone standardization program, CDC successfully conducts programs to standardize estradiol and vitamin D, and it works with PATH to develop new programs, promote accurate testing, and establish reference ranges.
3:50 PM
4:15 PM
Measurement and Normalization of Low-abundance Protein Analyte Panels in Dried Blood Spots (DBS) Using an Automated SISCAPA Workflow
Leigh Anderson
SISCAPA Assay Technologies
Long Abstract | Financial Disclosure
An improved high-throughput automated SISCAPA workflow was devised and applied to dried blood spots samples (DBS). This protocol provides significant sensitivity and throughput improvements for protein measurement in DBS, and overcomes sample hematocrit and volume variability issues that can limit the accuracy of protein-based DBS technology in clinical settings. We measured two 11-protein panels using 3-minute cycles, including targets in the low ng/ml range, and normalized results by separately measuring a set of high-abundance proteins comprising 80% of the total plasma protein, as well as the dominant red cell proteins, to yield surrogate hematocrit and total plasma load.
Mass Spectrometry vs Designer Synthetic Cannabinoids
Marilyn Huestis
National Institute on Drug Abuse
Long Abstract | Bio
Identification and quantification of new designer drugs is an important public health and safety issue; however, the metabolism of synthetic cannabinoids and appropriate urinary target analytes are frequently unknown. With more than 40 entirely new designer drugs identified by the US Drug Enforcement Agency in the first five months of 2013, designer drugs are the emerging face of drug abuse. We utilized LC-MS/MS to confirm and quantify metabolites, optimize cutoffs for synthetic cannabinoid immunoassays and, when metabolic profiles were unknown, human hepatocyte incubation and high resolution mass spectrometry to decipher urinary targets.
Targeted Metabolomics Identifies Glutathione Inhibition as a Mode of Sensitizing Drug-resistant Ovarian Cancer Cells to Nutrient Deprivation by L-asparaginase
Preeti Purwaha
Department of Bioinformatics and Computational Bio
Long Abstract | Bio
L-asparaginase (L-ASP) is used for treatment of acute lymphoblastic leukemia. Our recent metabolomic investigations showed that cell lines, such as OVCAR-8 which exhibited sensitivity towards L-ASP are glutamine addicted while drug resistant cells, such as OVCAR-4 are glutamine independent. To investigate the biological pathways associated with glutamine independence, we profiled the metabolites of TCA cycle, pentose phosphate pathway, nucleotides, and GSH/GSSG using a label-free, HILIC-based LC-MS/MS assay. Results revealed interesting differences in GSH metabolism between OVCAR-4 and OVCAR-8 cells. Treatment of OVCAR-4 cells with L-ASP combined with BSO, an inhibitor of GSH synthesis, induced cytotoxicity in these cells.
Standardization of Immunosuppressant Drug Measurements
Thomas Annesley
University of Michigan
Long Abstract | Bio
The movement of LC–MS/MS into clinical laboratories has been accelerated by the need to monitor immunosuppressant drugs. The analytical sensitivity and specificity of LC-MS/MS allows laboratories to quantify cyclosporine, tacrolimus, sirolimus, everolimus and mycophenolic acid, often with multiple drugs quantified within a single run. Yet even for small drug molecules the path to standardization has not been easy, as shown by calibrator traceability studies, proficiency testing schemes and inter-laboratory comparisons. This presentation will cover the state of standardization of immunosuppressant quantification from the perspective of both immunoassay and LC-MS/MS, each of which has had challenges to success.
4:15 PM
7:00 PM
RECEPTION
@ Exhibit Hall

Enjoy mingling with colleagues and Exhibitors. Take time to explore the Posters, which will be attended from 5:00 - 6:00 PM. Heavy Appetizers and Drinks to be provided.

POSTER PRESENTERS: If you are presenting a poster from 5-6 PM your poster should've been up at 8 AM.

Sponsored by: TBA
5:00 PM
6:00 PM
POSTERS
@ Exhibit Hall

Group 'b' Posters to be attended from 5:00 - 6:00 PM

All Posters should be removed by 7:30 PM.
TRANSLATIONAL MASS SPEC PLENARY LECTURE
@ Harbor Ballroom
Chair: David Herold
7:00 PM
7:45 PM
Genetic Defects in Bile Acid Synthesis Causing Liver Disease - Diagnosis and Treatment - Translational Medicine from Mass Spectrometry Discovery to the Bedside
Kenneth D. R. Setchell
University of Cincinnati College of Medicine
Long Abstract | Financial Disclosure
The combined application of FAB-MS with metabolic profiling using GC-MS, and electrospray HPLC-MS with accurate mass measurement led to the discovery of six genetic defects in the complex pathway for bile acid synthesis from cholesterol. The biochemical basis of each defect was characterized, mutations in genes encoding the respective enzymes confirmed, and a successful therapy developed. These genetic defects accounted for 2% of >12,000 cases screened for unexplained liver disease. Early diagnosis is important because patients with these fatal forms of metabolic liver disease respond successfully to oral primary bile acid therapy, thereby avoiding the need for liver transplantation - the only other therapeutic option. Based on the successful application of this therapeutic strategy, oral cholic acid therapy was granted approval in November 2013 by the European Medicines Agency (EMA) for the treatment of these bile acid synthetic defects and is currently under review by the US FDA.

This presentation of more than two decades of work is an example of translational medicine - taking a clinical problem at the bedside, to the utilization of mass spectrometry to define a new class of metabolic liver disease, to the development of a successful and safe therapy, and finally taking this back to the bedside by obtaining regulatory approval.

MS to Clinic Presentations
@ Harbor Ballroom
Chair: Ken Setchell
Sponsored by:
Thermo
7:45 PM
Alpha-1-Antitrypsin Deficiency Diagnostics; the Journey from a Research Core to the Clinical Laboratory
H Robert Bergen, III & David L. Murray
Mayo Clinic
Long Abstract | Biography
α-1-antitrypsin deficiency is the result of a genetic disorder at two major loci. Correct diagnosis includes serum quantification and identification of the specific variant present. This is typically accomplished with multiple diagnostic tests including quantification (nephelometry), genotyping and/or phenotyping by isoelectric focusing. We unsuccessfully attempted analysis of intact α-1-antitrypsin by electrospray. Subsequent analysis of tryptic peptides containing the S and Z variants and a surrogate peptide for quantitation yielded a LC-MS/MS inclusive assay for both quantification and identification. We will detail the research and subsequent assay modifications which yielded a validated clinical test.
8:00 PM
LC-MS/MS Measurement of Estradiol in Routine Patient Care
Laura Owen & Brian Keevil
University Hospital of South Manchester
Long Abstract | Biography
We have developed a high throughput LC-MS/MS assay for estradiol and estrone measurement which has been in use for 6 months. During this time there have been 30 patients whose clinicians have sent samples for this LC-MS/MS analysis due to it superior sensitivity and specificity compared to immunoassay. It has been particularly of use in the breast cancer population and has identified immunoassay interference in 3 patients preventing any further investigations. The simultaneous measurement of estrone gives additional information towards calculating total estrogen status.
8:15 PM
Impact of Rapid Organism Identification via Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight Combined with Antimicrobial Stewardship Team Intervention
Angela Huang & Jerod Nagel
University of Michigan Health System
Long Abstract | Biography
Background. Integration of MALDI-TOF with antimicrobial stewardship team (AST) intervention has the potential for improvement in patient outcomes. Methods. A pre-post quasi-experimental study was conducted to analyze the impact of MALDI-TOF with AST notification and intervention following blood culture Gram stain, organism identification, and antimicrobial susceptibilities. Outcomes were compared to a historic control group. Results. 501 patients were included in the final analysis. MALDI-TOF with AST intervention decreased time to organism identification (84.0 vs 55.9h, P<.001), and improved time to effective (30.1 vs 20.4h, P=.021) and optimal antibiotic therapy (90.3 vs 47.3h, P<.001). Mortality, length of ICU stay, and recurrent bacteremia were lower in the intervention group. Acceptance of an AST intervention was associated with a trend toward reduced mortality (OR 0.55,P=.075).
8:30 PM
10:00 PM
HOSPITALITY
@ Shoreline Patio

Enjoy the San Diego evening down by the Marina with heaters and fire pits. Drinks provided. At Shoreline Patio by the pool.
8:30 PM
10:00 PM
YOGA & MEDITATION
@ Nautilus 5

Yoga is a complimentary offering for all MSACL registrants.
Participants welcome at anytime during the class.
A limited number of yoga mats will be 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. Space is limited.
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 presenters for Tuesday must have their posters placed by the end of breakfast (8 AM).
7:00 AM
8:00 AM

CORPORATE WORKSHOPS (AM)

Phytronix
Marina 6

Accelerating sample analysis in clinical and forensic toxicology
Dr. John Laycock, Amgen and Dr. Pierre Picard, Phytronix

Pre-Register

This workshop is aimed at providing information in regards to new methods that will increase your sample analysis speed. Our speakers will offer valuable insight and share with you new ways to improve your workflows using LDTD-High Resolution MS and new developments in forensic toxicology using the LDTD Ion Source. We will present a validated method for the analysis of 13 benzodiazepines using LDTD-MS/MS in under 10 seconds per sample. We will discuss elements of sample preparation and the analysis steps of the LDTD. We will also compare cross-validated data with gold standard analysis method by LC-MS/MS. All compounds are analyzed simultaneously in 6 seconds per sample.

Spark Holland
Seabreeze

Automated dried blood spot analysis with FTDTM-SPE-MS/MS – no punching, no hematocrit issue
Emile Koster and Bert Ooms

Quantitative dried bloodspot analysis may suffer from variations in hematocrit (Ht) causing unacceptable inaccuracy and imprecision. Full-spot desorption avoids Ht effects on spot size, but Ht effects on analyte recovery may still remain. The best strategy to reduce this effect is to maximize the recovery of analytes from a dried blood spot. Flow Through Desorption (FTDTM) technology with solvent heating appears to be very effective, especially for immunosuppressant drugs. A completely automated DBS method based on SPE–MS/MS analysis, independent of the blood Ht value, will be presented. The DBSA is a front-end for any LC–MS/MS system and has card capacity from 96 (standard) to ~500 (extended). A built-in camera provides sample tracking, spot locating, and recording of the desorbed spot area. Both full-spot and partial-spot analysis are supported.

Phenomenex
Spinnaker

Chromatographic Method Development for the LC/MS User
Seyed Sadjadi, Senior Scientist

Pre-Register

The advent and rapid popularity of the MS detectors have led to shorter and narrower columns and subsequently faster chromatographic run times. This presentation covers a brief discussion of chromatographic media, bonded phases and method development in connection with LC/MS and LC/MS/MS technology and requirements.

PLENARY LECTURE SERIES
@ Harbor Ballroom
Chair: Andy Hoofnagle
8:30 AM
9:15 AM
Accelerating Clinical Mass Spectrometry; Workflows and Whitespace
Russell Grant
Laboratory Corporation of America
No Long Abstract | Biography | Financial Disclosure
Over the last decade, LC-MS/MS technologies have evolved from their innate promise of reference level measurement to become a circumscribed technology in patient management. The implementation of LC-MS/MS reflects the abstruse and inchoate status of the platform; we are challenged with ensuring future advancements are not ersatz as we balance sanguine hype and egregious hubris. This presentation will highlight our efforts to enable LC-MS/MS and peripheral technologies in clinical diagnosis, subsequently converting the apostates. Specific examples involving contiguous automation from patient to result, atavistic enhancements in hormone analysis, targeted protein analysis despite opprobrium and functional immune-response determination will be shown as we adduce the locution "mass spectrometry est sui generis".
9:15 AM
10:00 AM
Moving Mold Identification into the 21st Century with MALDI-TOF MS
Anna Lau
National Institutes of Health
No Long Abstract | Biography
MALDI-TOF MS has revolutionized clinical microbiology laboratories, enabling accurate bacterial and yeast identification within minutes at minimal costs and labor. However, mold identification by mass spectrometry has lagged, forcing many laboratories to rely on slow phenotypic methods that are prone to misidentifications. This presentation will describe the development of the NIH Mold Database - a clinically comprehensive, freely available database for the identification of molds from solid media. Its clinical performance will be highlighted through a series of multicenter studies and case presentations, with the NIH Mold Database outperforming a commercially available database. The NIH Mold Database has pushed the boundaries of clinical mycology, providing faster therapeutic guidance, positively impacting patient care, and opening new doors and challenges for clinical research.
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
Clinical Use of LC-MS/MS for Protein Analysis
Chair: Brian Keevil
Track 2
Harbor Ballroom 2
New Solutions to Nagging Problems
Chair: Dan Holmes
Track 3
Harbor Ballroom 3
Microbiology II
Chair: Susan Butler-Wu
Track 4
Marina 6
Basics: Implementing Small Molecule Assays
Chair: Nandu Chindarkar
10:45 AM
11:10 AM
Using the Accurate Molecular Mass of Monoclonal Light Chains to Detect Residual Disease in Patients with Multiple Myeloma
John Mills
Mayo Clinic
Long Abstract | Bio
Multiple myeloma is a plasma cell proliferative disorder where the M-protein (monoclonal immunoglobulin) serves as a surrogate marker for the plasma clone responsible for its production. In multiple myeloma along with clinical symptoms, the diagnosis, monitoring and treatment decisions are dependent on the detection of an M-protein. The absence of an M-protein is indicative of remission, whereas an increase of M-protein can signal progression. Here, we report a mass spectrometry based method to determine the accurate molecular mass of the M-protein in patients diagnosed with multiple myeloma. We are able detect these patient-specific M-proteins in sera months after successful therapy when the patient was deemed in remission using stringent criteria. This highlights the enhance sensitivity mass spectrometry can offer in detecting monoclonal immunoglobulins in patients sera.
Reducing the Need for Calibration Standards
Geoffrey Rule
ARUP Laboratories
Long Abstract | Bio
Mass spectrometers are extremely reliable tools for clinical testing, especially when isotopically labeled internal standards are used. This lead us to consider if a traditional number of calibration standards are always necessary. With linear data, a single calibrator should be sufficient to establish detector response for quantitation. Here we compare efforts utilizing a weighted historical response factor to a conventional approach. For nonlinear data we utilize a unique solution accounting for isotope contributions to internal standard. In this case, use of one or two experimentally determined constants and a single adjustable parameter still allows use of a single calibration standard.
Multicenter Evaluation of MALDI-TOF MS for the Identification of Bacteria and Yeast
Christopher Doern
Virginia Commonwealth University Medical Center
Long Abstract | Bio
Inter-laboratory performance of MALDI-TOF MS for organism identification is unstudied. Herein, we assess Bruker Biotyper MALDI-TOF MS validation data from > 5,500 isolates from 9 medical centers. Of 1,820 Gram positives, 94.8% were correctly identified, 1.9% incorrectly identified while 3.2% could not be identified. Of 2,635 Gram negatives, 95.9% were correctly identified, 2.3% incorrectly identified while 1.8% could not be identified. Of 382 yeast, 95.8% were correctly identified, 2.4% incorrectly identified. Of 863 anaerobes 92.78% were correctly identified, 0.1% incorrectly identified while 8.1% could not be identified. In conclusion, MALDI-TOF MS is reliable and reproducible for organism identification across laboratories.
Vitamin D by LC-MS/MS: Practical Tips and Stories from the Front-Line
Lorin Bachmann
Virginia Commonwealth University
No Long Abstract | Bio | Financial Disclosure
Are you thinking of measuring a small molecule such as Vit D by mass spectrometry? Maybe you have an instrument and are wondering how to bridge the [large!] gap between installation and launching a validated method? Want some advice on how to deal with problems encountered along the way? This session is intended to provide practical advice, examples and first-hand accounts of experiences encountered during development, validation and on-going performance assessment of Vitamin D testing by LC-MS/MS. Method selection/development tips, validation design examples and troubleshooting case studies will be presented.
11:10 AM
11:35 AM
Meeting the Needs of In-house Thyroid Cancer Patients: Identifying Appropriate Thyroglobulin and Thyroglobulin Antibody Testing Algorithms
Sarah Wheeler
University of Pittsburgh
Long Abstract | Bio
Thyroglobulin (Tg) immunoassay methods are susceptible to Tg antibody (TgAb) interference necessitating referral of high TgAb specimens. The most common referral tests are radioimmunoassay (RIA) and LC/MS/MS. This study will identify the limitations of UPMC in-house Tg to develop appropriate testing algorithms. High TgAb specimens (n=20) from adults post total thyroidectomy were analyzed for Tg using immunoassay (DxI800; Beckman Coulter), RIA (USC Endocrine Laboratories), and LC/MS/MS (Quest Diagnostics). At low Tg concentrations there was poor correlation between the DxI800 and other methods (RIA, R2 = 0.64; LC/MS/MS, R2 = 0.76). Comparison of RIA and LC/MS/MS showed some discrepancy (R2 = 0.82) but had high correlation for low Tg concentrations (<13 ng/mL; R2 = 0.96). Reference testing using LC/MS/MS for TgAb positive specimens that exceed 13ng/mL Tg is clinically necessary.
Finding the Fakes of Drug Testing; New Strategies to Identify Substituted Samples
Gregory Janis
MedTox Laboratories
Long Abstract | Bio | Financial Disclosure
Urine- based drug testing is a key component of pre-employment screening, probationary monitoring, and pain management compliance programs. There exists an enormous incentive for deception if a sample donor believes they are at risk of failing. Substituted synthetic urine samples are the most common deception attempted today. Synthetic urine products are increasingly complex, capable of passing routine sample validity tests. We have characterized synthetic urine and mined natural urine identifying novel targets to distinguish real from artificial samples. Exogenous markers and esoteric endogenous markers, functioning as validity indicators, have been incorporated into LC-MS/MS assays at high risk for substitution.
Comparison of Preparation Methods and Instrumentation Platforms for MALDI-TOF MS Identification of Medically Relevant Yeast Species
Morgan A. Pence
Washington University in St. Louis
Long Abstract
Our objective was to evaluate multiple “direct smear” techniques for yeast identification using the VITEK MS and Bruker Biotyper MALDI-TOF MS platforms. Yeast isolates (117) were analyzed after 24 and 48h incubation. Isolates were analyzed by the direct smear method +/- a 25% formic acid on-plate extraction on both platforms and with 100% formic acid on Biotyper. VITEK MS correctly identified 113 isolates at 24h with one misidentification. With a revised threshold of >1.7, Biotyper correctly identified 103 isolates, with 3 misidentifications. Formic acid significantly increased the identification rate. MALDI-TOF MS analysis for yeast identification drastically improves time to identification.
How to Make a Clinical LC-MS Testosterone Assay that Actually Works for Real Patients: Lessons Learned from the Front Lines
Nigel Clarke
Quest Diagnostics Nichols Institute
Long Abstract | Financial Disclosure
This presentation will discuss the whole process of developing, validating and operationalising a Dx MS assay using an example testosterone assay currently being offered in our lab. Discussion of the approaches, instrumentation and methodologies for analysis of small molecules will be examined. Examples will be given of the sample preparation strategies, analysis modes and software options as well as how they behaved in our laboratory.
11:35 AM
12:00 PM
Shotgun Proteomics-based Clinical Testing for Diagnosis and Classification of Amyloidosis
Surendra Dasari
Mayo Clinic
Long Abstract | Bio
Shotgun proteomics technology has matured in research laboratories and is poised to enter clinical laboratories. However, the road to this transition is sprinkled with major technical unknowns like long-term stability of the platform, reproducibility and clinical utility over traditional IHC platform. As a result, diagnostic laboratories have avoided using shotgun proteomics for routine diagnostics. We developed two shotgun proteomics assays (FFPE tissue-based and subcutaneous fat aspirate-based) for amyloid subtyping by standardizing the platform for better quality control and earning clinical acceptance. These assays have >91% success rate of subtyping compared to 48% of traditional IHC technique. These assays were utilized to detect 19 different amyloid subtypes in 5600 patients. Retrospective data mining of amyloid proteomes revealed ALect2 as a new third major type in the kidney.
Drug Screening Using Multiple Methods (immunoassay, GC-MS, LC-MS/MS): Is there an Easier Way?
Kara Lynch
University of California San Francisco
Long Abstract | Bio | Financial Disclosure
In the clinical laboratory, drug screening is done by a combination of methodologies. Recently, drug screening methods that utilize high resolution mass spectrometry(HRMS) have been published, however, few studies have compared HRMS screening to traditional drug screening methodologies. The objective of this study was to develop LC-HRMS and LC-MS/MS drug screening methods that are identical except for the mass spectrometer used and to compare them to immunoassay screening for their ability to identify drugs/metabolites in routine specimens. The MS methods were more sensitive and specific compared to immunoassay and had a better positive and negative predictive value. They were also capable of detecting additional compounds with a high degree of confidence in positive identifications.
Identification of Filamentous Fungi Using MALDI-TOF Mass Spectrometry
Alexander Gertel
Medical College of Wisconsin
Long Abstract | Bio
Identifying filamentous fungi using MALDI-TOF MS has been difficult and spectral libraries have been inadequate for generating consistent matches. This study aims to verify this application of MALDI-TOF using a liquid broth-based growth protocol. Additionally, we hypothesize that advances in a current mold spectrum library will yield more frequent matches for identification. MALDI-TOF analysis of 158 mold isolates matched microscopic identification 72% of the time and DNA sequencing proved MALDI-TOF to be accurate in 82% of total cases. MALDI-TOF following growth in liquid broth is a viable method for identifying molds in the clinical laboratory.
Development and Validation of Broad Spectrum Drug Screening Method on a Time-Of-Flight Mass Spectrometer
Nandkishor Chindarkar
University of California- San Diego
Long Abstract | Bio | Financial Disclosure
Recent developments in the high resolution mass spectrometry have lead to instruments with improved sensitivity and specificity as compared with contemporary low resolution LC-MS/MS instruments. HRMS instruments have potential to be universal detectors for broad spectrum drug screening in clinical and toxicology laboratories. We investigated a panel of more than sixty compounds in urine with a dilute, hydrolyze, and shoot methodology using a TOF instrument. Qualitative results obtained by our method had very good correlation with those obtained by LC-MS/MS methods. We observed that the addition of a fragment ion was essential in eliminating false positive results. This report will outline the challenges faced during the method development for a broad spectrum drug screen using high resolution TOF MS.
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.
• Poster Group 'a' to present during the 2:00 - 3:00 PM coffee break in Exhibit Hall.
1:00 PM
2:00 PM

CORPORATE WORKSHOPS (PM)

Phenomenex
Harbor Ballroom 1

Solid-Phase Extraction: Tools, Tips and Tricks on How to Develop the Best SPE Method
Seyed Sadjadi, Senior Scientist

Pre-Register

Solid Phase Extraction (SPE) presents a versatile and reliable sample preparation procedure in many clinical laboratories. This workshop presents an in-depth discussion of the various SPE support media, bonded ligands and their interaction mechanism. Additional tools and guideline will be provided to aid basic and successful SPE method development.

SimulTOF
Harbor Ballroom 2

Accurate determination of protein profiles in complex biological samples by MALDI-TOF MS
Marvin L. Vestal, SimulTOF Systems

Protein profiling has been investigated for various diagnostic applications such as cancer detection, biomarker determination and pathogen identification (approved by the FDA last year). Despite enormous effort and expense, biomarkers for other diagnostic applications have not been approved. The difficulties inherent in correlating the mass spectrum of a complex mixture with a biological state are well-known. These include variations in sample acquisition, processing, and preservation; reproducibility of mass spectrometric data acquisition and processing; and unrelated biological variations. SimulTOF’s focus is minimization of instrumental variations. The SimulTOF 100 Linear MALDI-TOF, operating at laser rate of 5 kHZ, provides high sensitivity and reproducibility. Use of this technology on biological fluids, with and without purification, as well as in tissue imaging, will be presented. In addition, we will present a new approach to spectral comparison.

AB SCIEX
Harbor Ballroom 3

(1) LC/MS/MS sample prep automation, workflow integration and data flow for the clinical research laboratory
(2) Resolution of Lipid Biomarkers using Differential Mobility Separation

(1) Michael Jarvis, Technical Marketing Manager, Clinical Research - AB SCIEX
(2) Paul RS Baker, PhD, Senior Applications Specialist, Global Lead Scientist in Lipidomics - AB SCIEX

No summary submitted.

Bruker Daltonics
Marina 6

Recent Developments in the MALDI Biotyper for Microbial Identification and Beyond and Development of a SISCAPA MALDI Workflow for Biomarker Development and Routine Measurement
Gongyi Shi, Director of Scientific Affairs, Bruker Daltonics and N. Leigh Anderson, SISCAPA Assays Technologies Inc.

Routine identification of microbes using MALDI-TOF mass spectrometry is widely accepted. Recent developments in microbial identification, such as the FDA clearance for IVD use in the US, and development work on the use of the MALDI Biotyper platform for the investigation of antibiotic resistance will be presented. SISCAPA MALDI workflows for specific assays of protein biomarkers have been shown to be robust and highly quantitative. An automated platform for sample preparation and data analysis for SISCAPA MALDI will be presented, and its simplicity and throughput advantages discussed.

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

Group 'a' Posters to be attended from 2:00 - 3:00 PM.
GENERAL SCIENTIFIC SESSION 4
Track 1
Harbor Ballroom 1
LC-MS/MS Protein Analyis Performance - A Critical View
Chair: Stephen Master
Track 2
Harbor Ballroom 2
Applications of Accelerator-MS and ICP-MS
Chair: Paul Jannetto
Track 3
Harbor Ballroom 3
Metabolomics II: Clinical Diagnostics
Chair: Gary Patti
Track 4
Marina 6
Basics: Automating Data Exchange
Chair: Judy Stone
3:00 PM
3:25 PM
Unraveling Trypsin Digestion of Apolipoproteins A-I and B to Evaluate the Impact on Imprecision of MS-based Quantification
Irene van den Broek
Leiden University Medical Center
Long Abstract | Bio | Financial Disclosure
Trypsin digestion of apolipoproteins A-I and B was evaluated in normo- and hypertriglyceridemic serum to characterize the cleavage efficiency of multiple signature peptides. Addition of stable-isotope labeled peptides prior to digestion enhanced reproducibility, and only minor differences were observed across the various specimens. The total imprecision of apolipoprotein A-I and B measurement by LC-MS/MS, after quadruplicate sample preparations on five separate days, was <10% for all measured peptides. In general, 70 to 90% could be attributed to variations before LC-MS/MS analysis. The results provide valuable information about the impact of proteolysis on apolipoprotein A-I and B measurement imprecision, of particular importance for application in clinical chemistry.
Accelerator Mass Spectrometry to Detect Platinum-induced DNA Adducts and Identify Chemoresistance: from Bench Research to a Phase II Clinical Trial
Paul Henderson
University of California Davis
Long Abstract | Financial Disclosure
Accelerator mass spectrometry (AMS) can measure carbon-14 at the sub-attomole level in mg-size specimens, and is commonly used for radiocarbon dating. We use AMS to measure DNA adducts induced by platinum chemotherapeutic agents. As alkylating agents, platinum agents kill cancer cells through induction of DNA adducts. We hypothesize that cancer cells with low platinum-induced DNA adducts will survive chemotherapy and are chemoresistant. Pre-clinical studies in cell lines and animal models support this hypothesis. After completion of pre-clinical studies and a Phase I clinical trial, a Phase II trial is going on and will be presented at the meeting.
Targeted Profiling of Inflammation Related Oxylipins in Human Biofluids: Application to Surgery Patients
Arnaud Wolfer
Imperial College London
Long Abstract | Bio
Current clinical risk scores are inadequate for surgical outcomes prediction, and robust biomarkers for the early detection of complications are lacking. To help understand these conditions, a sensitive UPLC-MS profiling method for the quantification of 48 oxylipins involved in inflammation signalling has been developed. Blood samples of patients undergoing surgery were analysed and significant lipid mediator evolutions were related to different stages of the inflammatory response and invasiveness of the intervention. Serial oxylipin measurements are of clinical merit in surgery and may serve as a novel biomarker for prediction of poor surgery outcomes, by accounting for patient-specific pre-operative inflammatory states.
Translating Between IT and MS Subcultures - An Overview of IT Technologies and Terminology Designed for Clinical Laboratory Personnel
Tony DaSilva
Kaiser Permanente
Long Abstract | Bio
In the quest to automate mass spectrometry instrument interfaces in the clinical laboratory, IT technologies play a critical role in the way disparate systems integrate. At play are a wide range of networking technologies, a dizzying array of communications protocols, laboratory information systems, middleware systems, and challenging instrument software systems. In most cases, laboratorians are the ones needing to orchestrate how all this technology must play together and to figure out what kind of assistance they need from their IT departments, vendors, and technical staff for a successful implementation. This presentation will provide laboratorians with the basic skills and understanding of the underlying IT technologies to effectively communicate with their technology providers.
3:25 PM
3:50 PM
Development of a High-Throughput Mass Spectrometry Method to Screen for Familial Hypercholesterolemia and Detect a Common Genetic Variant of Apolipoprotein B
Jeff Meeusen
Mayo Clinic
Long Abstract | Bio
Familial hypercholesterolemia (FH) is an autosomal dominant genetic disorder largely underdiagnosed (prevalence 1:500), resulting from mutations in LDLR, APOB or PCSK9 genes and characterized phenotypically with elevated low-density lipoprotein cholesterol and apolipoprotein B particles. We developed an LC-MS/MS method in patient specimens utilizing peptides specific to apolipoprotein B (apoB100) and the most common APOB mutation (R3500Q). Method optimization was achieved by reduction in trypsin digestion times without compromising signal intensity and accuracy. Identification of an R3500Q mutation on LC-MS/MS was confirmed with genetic testing. Our data supports LC-MS/MS as a promising FH screening method, particularly in pediatric populations.
Reduced Sample Volume and Simplified Sample Preparation in the Simultaneous Measurement of Zinc, Antimony, Bismuth, and Manganese in Whole Blood Using ICP-MS
Carrie Haglock-Adler
ARUP Institute for Clinical & Experimental Path
Long Abstract | Bio
To achieve required sensitivity from whole blood samples with Inductively Coupled Plasma-Mass Spectrometry (ICP-MS), historic extraction methods often required a time-consuming chemical and/or microwave digestion. The reported method describes a simplified dilution for the simultaneous analysis of Zn, Sb, Bi, and Mn in whole blood. A 100 uL sample was extracted with 4.9 mL of basic diluent containing an Indium internal standard. Goat blood was added to all calibrators to achieve a matrix matched method. Inter and Intra-assay imprecision was less than 5% for all four elements.
Validation and Application of a LC-HRMS Platform for Simultaneous Absolute Quantification and Biomarker Discovery in the Clinical Laboratory
Ilya Gertsman
University of California, San Diego
Long Abstract | Bio
We have recently developed and validated a Q-TOF mass spec platform that simultaneously collects high resolution/high mass accuracy data for discovery work while simultaneously performing absolute quantification of over 50 relevant analytes commonly measured for variety of inherited metabolic disorders. These include amino acids, organic acids, purines/pyrimidines, and acylcarnitines. Previously, multiple forms of instrumentation were required to measure all of these different chemical classes, but advances in both chromatography and the linear dynamic range of high resolution mass specs make this attainable with a singe system, We show this method in its application to a drug study involving Alkaptonuria patients.
A Road Map to Interfacing Your Mass Spectrometer
Judy Stone
CALM, Univ. of Calif., San Diego
Long Abstract | Bio
A barrier to implementing mass spectrometry (MS) in the clinical laboratory is the challenge of interfacing MS and automated sample preparation instruments to laboratory information systems (LIS) for electronic order and result transmission. This presentation will review basics of interface terminology, protocols, and workflows and will explain common differences encountered between interfacing mass spectrometers versus automated chemistry analyzers. Interfacing options now available from MS vendors will be discussed. Case histories will be used to illustrate recommendations to laboratorians for choosing between interface protocol options (.txt export, XML export, HL7), designing MS data storage architectures, selecting MS fields to export, interface testing and version control, and using middleware/LIS rules to automate data review.
3:50 PM
4:15 PM
What Contributes to Inter-laboratory Variability of Targeted Protein LC-MS/MS Assays: A Case Study Using IGF-1
Andy Hoofnagle
University of Washington Medical Center
Long Abstract | Bio | Financial Disclosure
The use of targeted LC-MS/MS assays for proteins will simplify the harmonization and standardization of clinical measurements. Careful investigation into the variability of the bottom-up proteomics process and its effect on inter-laboratory agreement is lacking. We developed a detailed standard operating procedure to quantify serum IGF-1, which is a well-characterized biomarker of growth hormone abuse, and deployed it in 5 laboratories in 3 countries. We identified the variability due to instrumentation and the influence of the approach to calibration on inter-laboratory agreement. The primary source of variability was not derived from the sample preparation but from the method of calibration.
Use of Isocratic Pump Direct Injection and Inductively Coupled Plasma-Mass Spectrometry for Analysis of Lead in 20 µL of Whole Blood
Sarah Hackenmueller
University of Utah/ARUP Laboratories
Long Abstract | Bio
Lead in whole blood is currently analyzed to monitor exposures in children and in the workplace. Routine analysis by inductively coupled plasma-mass spectrometry (ICP-MS) utilizes a peristaltic pump for sample introduction into the ICP-MS, requiring 1-2 mL of prepared sample for each injection, despite the use of discrete injection valve systems. We investigated the use of an isocratic pump for discreet 100 µL injections into the ICP-MS. This method modification resulted in an 80% reduction in required sample and reagent volumes while maintaining acceptable analytical sensitivity, imprecision, and accuracy with respect to the original assay.
Lipidomics of Omega-3 Transgenic Mice Reveals a Panel of Anti-inflammatory and Pro-resolving Mediators
Giuseppe Astarita
Waters Corporation
Long Abstract | Bio | Financial Disclosure
A large number of studies have demonstrated reduced disease risk and health benefits of long term omega-3 fatty acid supplementation. In this study, we used high-throughput MS assays, including a prototype microfluidic MS platform, to characterize the molecular phenotype of plasma samples from an animal model of long term omega-3 supplementation. Our results highlight a novel panel of anti-inflammatory and pro-resolving mediators, which may be involved in the health benefits associated with alterations of the omega-6/omega-3 fatty acids ratio.
Keeping Techs from Early Retirement: Approaches to Data Review in Multiplexed Mass Spectrometry Assays
Jane Dickerson
Seattle Children's Hospital
Long Abstract | Bio
Development of multiplexed mass spectrometry assays seems like every lab’s dream – however, increasing the number of analytes directly increases the complexity of the analysis. In an assay that measures 20 opioids and metabolites, there are nine metrics monitored for each of four sample preparation methods per analyte, totaling 720 data points per patient specimen. Without automated software tools, result preparation involved several manual processes and decision-making steps requiring extensive technologist time. Implementation of either custom or commercial software to perform quality control calculations reduces the amount of time required to manually review data, and ultimately, reduces the risk for errors.
4:15 PM
7:00 PM
RECEPTION
@ Exhibit Hall

Enjoy mingling with colleagues and Exhibitors. Take time to explore the Posters, which will be attended from 5:00 - 6:00 PM. Heavy Appetizers and Drinks to be provided.

Sponsored by: TBA
5:00 PM
6:00 PM
POSTERS
@ Exhibit Hall

Group 'b' Posters to be attended from 5:00 - 6:00 PM

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

USER GROUPS
@ Rooms off Harbor Island Foyer
Commercial Presentations for current and future product users. Note: Access Controlled by Sponsor.

SimulTOF
Harbor Ballroom 1

MALDI Imaging Mass Spectrometry: Technical Advances and Clinical Problem Solving
Jeremy L. Norris, National Research Resource for Imaging Mass Spectrometry,Vanderbilt University School of Medicine

MALDI Imaging Mass Spectrometry (IMS) is a unique technology that combines the spatial advantages of microscopy with the chemical specificity of modern mass spectrometers. The combination of these advantages can be leveraged to investigate biological systems of clinical importance (e.g., tissues). Recent advances in MS instrumentation, sample preparation methods, and biocomputational approaches to image analysis are converging to enable novel, clinically important applications of IMS. Advances emerging from collaboration between the National Research Resource for Imaging Mass Spectrometry, Department of Biochemistry, Vanderbilt University School of Medicine and SimulTOF Systems will be discussed by key personnel from both organizations.

Thermo Scientific
Harbor Ballroom 2

New Application Seminar - Mass Spectrometric Immunoassays (MSIA) - The evolution of targeted MS workflows and platforms
Dr. Eric Niederkofler, Dr. Lewis Couchman, Dr. Benoit Coulombe, Dr. Chegjie Ji

Pre-Register

Speakers will describe the applications & advantages of Thermo Scientific MSIA (Mass Spectrometric Immunoassay) workflows. MSIA workflows are comprehensive and customizable targeted-MS based approaches that combine highly selective sample interface preparation with MS for simultaneous qual/quan analysis. Dr. Niederkofler will define the MSIA workflows and technologies. Dr. Couchman will highlight reproducible and high-throughput methods for intact insulin and its therapeutic analogs from donor samples by using MSIA with high-resolution accurate mass (HRAM) MS instrumentation. Dr. Ji will discuss a new approach to detect low-level protein, TNFa. The speakers will participate in a panel discussion, hosted by Dr. Coulombe, to provide further insight into MSIA applications and how new users can quickly adopt MSIA workflows in their labs.

AB SCIEX
Harbor Ballroom 3

(1) New sensitivity and selectivity gains with the QTRAP 6500® system and SelexION™ technology for steroid analysis and other clinical research applications.
(2) Innovations in microflow LC-MS to improve methods in clinical research and forensic toxicology applications.

1) Michael Jarvis, Technical Marketing Manager, Clinical Research - AB SCIEX
2) Evelyn McClure, MSc., Application Scientist - AB SCIEX

Bruker Daltonics
Marina 6

(1) Late-Breaking Updates from the MALDI Biotyper R&D group
(2) Clinical Impact of MALDI-TOF MS: Making MALDI Matter beyond the Lab
(3) GC-MS/MS: Applications for Routine Clinical and Emergency Toxicology

(1) Markus Kostrzewa, Ph. D. and VP for Clinical Research at Bruker Daltonics
(2) Christopher Doern, Associate Director of Microbiology at the VCU Medical Center
(3) Jennifer Colby, Ph. D., and Fellow of Clinical Chemistry and Toxicology at UCSF.

Join us for three short (about 15 minutes each) presentations by current Bruker users, followed by plenty of time Q&A, discussion and networking. All current Bruker users and those interested in Bruker Daltonics mass spectrometry products may attend. will present some late-breaking updates from the MALDI Biotyper R&D group. Prof. will speak on, “Clinical Impact of MALDI-TOF MS: Making MALDI Matter beyond the Lab.” And Jennifer Colby, Ph. D., and Fellow of Clinical Chemistry and Toxicology at the University of California, San Francisco will present on, “GC-MS/MS: Applications for Routine Clinical and Emergency Toxicology.”

8:30 PM
10:00 PM
HOSPITALITY
@ Shoreline Patio

Enjoy the San Diego evening down by the Marina with heaters and fire pits. Drinks provided. At Shoreline Patio by the pool.
8:30 PM
10:00 PM
YOGA & MEDITATION
@ Nautilus 5

Yoga is a complimentary offering for all MSACL registrants.
Participants welcome at anytime during the class.
A limited number of yoga mats will be 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. Space is limited.
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)

Indigo Biosystems
Marina 6

Software for Automating Data Review for Real World Assays
Randy Julian

Indigo Biosystems has produced the first automated chromatographic analysis software which allows targeted review and review by exception for MRM quantitation. In this workshop, the features of ASCENT that automate data review will be described, including Indigo's novel approach to peak integration. This presentation will review how ASCENT works and include examples of a number of real world chromatographic and mass spectrometric problems and they are handled by ASCENT. We will show how our patented algorithms impact real data and simplify data analysis.

Zef Scientific
Seabreeze

LC-MS/MS Troubleshooting and Best Practices
James Byrd, Zef Scientific, Inc.

The technique of LC-MS/MS is powerful and specific, but it comes with a unique set of challenges. If you are looking for ideas to increase instrumental uptime in your lab, come join us for breakfast; the speaker will share systematic troubleshooting strategies and anecdotes from over a decade of LC/MS customer support. Find out how to avoid common pitfalls and save time and money. Zef Scientific is one of the nation’s leading independent multi-vendor service providers, focused on chromatography and mass spec.

GENERAL SCIENTIFIC SESSION 5
Track 1
Harbor Ballroom 1
Tissue Imaging and Analysis
Chair: Jeremy Norris
Track 2
Harbor Ballroom 2
Novel Sample Prep and Automation: Accelerating Laboratory Workflows
Chair: Jason Sawyer
Track 3
Harbor Ballroom 3
Microbiology III
Chair: Chris Doern
Track 4
Marina 6
Basics: Metabolomics
Chair: Caroline Johnson
8:30 AM
8:55 AM
Molecular Analysis in 3D Using Imaging Mass Spectrometry and Randomised Compression Methods
Andrew Palmer
University of Bremen
Long Abstract
Development of 3D imaging mass spectrometry has provided a technique with great potential to determine molecular localisations throughout a tissue volume. By simultaneously mapping multiple molecules a new perspective of tissue function and disease alterations is obtained which can reveal the volumetric interactions of molecules. However, the increased data volume and complexity requires suitable algorithms for extracting meaningful information. In this work tissue volumes are automatically identified based on their individual spectral differences using automated spatial segmentation. This analysis provides a molecular profile of each tissue type as well as visualisation of the 3D structure. This analysis is greatly accelerated for 3D images by a data compression scheme based on randomised data sampling.
DEEM - A New Approach to Automated Method Development for LC-MS/MS Sample Preparation
Roland Geyer
Tecan Switzerland AG
Long Abstract | Financial Disclosure
The new automated method development process Direct Extraction Elution Measurement (DEEM) will be presented. Method development using the AC Extraction Plate is simplified by evaluating the three 'pipette and shake' steps simultaneously. Using a Tecan EVO, an array of spiked solutions were prepared with different pH values (acid, neutral and basic) and organic solvent percentages ranging from 5% to 100%. These are extracted and directly analyzed to determine the extraction efficiency. Percentage extraction is plotted against organic solvent content and pH values to determine suitable extraction, rinse and elution conditions. Examples of developed methods will be shown.
An Assessment of MALDI-TOF MS for the Identification of Infrequently Encountered Gram-negative Bacteria Unidentifiable by Conventional Methods
Tanis Dingle
University of Washington
Long Abstract
Gram-negative bacteria that cannot be identified by biochemical methods in our laboratory have traditionally been identified by 16S rRNA sequencing. Despite it's accuracy, this method is laborious, time consuming and costly. We compared the performance of 16S rRNA sequencing to MALDI-TOF MS for 65 Gram-negative isolates that could not be identified by conventional means. MALDI-TOF MS correctly identified 100% and 90.9% of isolates to the genus and species level, respectively. Additionally, MALDI-TOF MS provided species level identification for 7 of 10 isolates for which 16S rRNA sequencing could only provide a genus level or bacterial complex group identification. These results highlight the utility of MALDI-TOF MS in providing rapid and accurate identification to the species level for infrequently isolated Gram-negative organisms that are difficult to identify by traditional methods.
Experimental Designs for Metabolomics: Untargeted Does Not Mean Unplanned
Gary Patti
Washington University School of Medicine
No Long Abstract
The purpose of this educational course is to discuss the basic principles that investigators new to metabolomics should consider when designing untargeted studies. Factors to be discussed will include sample choice for comparison, size of cohorts analyzed, instrument(s) applied, and public resources available for data processing. Special attention will be devoted to optimizing experimental design to answer specific types of scientific questions. A list of minimum criteria will be recommended that should be in place prior to analyzing clinical samples. Strategies to deal with the challenges of studying clinical laboratory samples will be discussed, such as sample normalization, sample handling, extraction procedures, biological variation, and sample heterogeneity.
8:55 AM
9:20 AM
High Performance MALDI MS, MS/MS, and Multiplexed MS/MS Tissue Imaging
Boone Prentice
Vanderbilt University
Long Abstract | Bio | Financial Disclosure
Imaging mass spectrometry (IMS) has demonstrated the unique ability to provide untargeted and spatially resolved molecularly specific information. Recent advances in matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry instrumentation have provided for systems capable of high throughput, high sensitivity, and high specificity imaging. Specifically, the SimulTOF 300 Tandem Mass Spectrometer (SimulTOF Systems, Sudbury, MA) allows for multiple high resolution precursor ion selection tandem mass spectrometry (MS/MS) experiments to be performed in a single laser shot. The goal of this technology is to allow for targeted analyte MS/MS imaging as well as untargeted MS/MS discovery in tissues.
An improved method to quantify total cotinine and trans-3'-hydroxycotinine in urine using UHPLC/MS/MS, automated sample preparation and automated data analysis
Joseph Alexander
Centers for Disease Control and Prevention
Long Abstract | Bio
We developed and validated an automated high-throughput, low-cost, sensitive and selective method to analyze the nicotine metabolites total cotinine and hydroxycotinine in urine. Sample preparation was performed on a PerkinElmer Staccato System Robotics station. We used UHPLC coupled to an AB Sciex API 6500 triple quadupole mass spectrometer for peak separation and detection, and we used Indigo ASCENT software for automated peak integration and quality assurance screening. The limit of detection was 0.030 ng/mL for both analytes on a sample size of 0.2 mL. Our method will be used to measure secondhand smoke exposure in large population studies.
High Intra- and Inter-Laboratory Reproducibility of Mass Spectrometric-Based Identification of Bacteria and Yeasts
Lars Westblade
Washington University in St. Louis
Long Abstract | Bio
Data is scarce describing the intra-/inter-laboratory reproducibility of MALDI-TOF MS for identification of microorganisms. Our objective was to assess the performance of the VITEK MS® v2.0 system at three distinct sites for identification of bacteria and yeast. The study included 10 isolates analyzed by two operators at three sites on five different days, analysis of 300 'challenge'ť isolates and analysis of 1,089 “rare” isolates. All experiments demonstrated excellent reproducibility and analytical performance characteristics for identification of all organism types. MALDI-TOF MS-based microbial identification exhibits high intra-/inter-laboratory reproducibility and can tolerate multiple operators using multiple reagent lots on multiple instruments.
Data Processing and Analysis for Metabolomics
Gary Siuzdak
Scripps Research Institute
No Long Abstract
The purpose of this section of the metabolomic educational course is to discuss and demonstrate the basic principles behind metabolomic data processing and analysis. This presentation will cover how informatic technologies allow for metabolomic data processing, including automated feature detection, retention time correction, alignment of chromatograms, peak area calculations and, ultimately, statistical analysis and identification of significantly altered features. This presentation will be geared to the non-expert, providing an overview of the most straightforward approaches to performing mass spectrometry-based metabolomic experiments.
9:20 AM
9:45 AM
Simultaneous Profiling of Multiple N-linked Glycans on FFPE Tissue Slides and Tumor Tissue Microarrays Using MALDI Mass Spectrometry Imaging
Richard Drake
Medical University of South Carolina
Long Abstract | Bio
A new MALDI imaging mass spectrometry workflow for simultaneous analysis of multiple N-linked glycans in formalin-fixed tissue microarrays (TMA) and standard FFPE tissue slides will be presented. Following antigen retrieval processing and on-tissue digestion with peptide N-glycanase, 30-50 glycan species can be detected. The approach is amenable to any FFPE tissue, and represents an additional molecular correlate assay for use with the TMA format. Additionally, depending on the construction of the TMA and targeted tumor type, the approach has the potential to create novel glycan biomarker panels for cancer detection and prognosis.
Deproteination of Serum Samples for LC-MS/MS Analyses Applying Magnetic Micro-particles
Michael Vogeser
Hospital of the University of Munich, Germany
Long Abstract | Bio
A novel, micro-particle based approach for sample preparation in LC-MS/MS was evaluated. In a commercially available kit, a proprietary protein denaturation reagent is combined with ferromagnetic particles which capture and remove denatured proteins from the sample. The principle was tested for amiodarone as an exemplary analyte. On-line solid phase extraction was applied as a second clean-up step. This set-up was found highly efficient and convenient. The principle competes with protein precipitation and seems highly attractive for automation, since no centrifugation or application of positive or negative pressure is required.
VITEK Mass Spectrometry for the Identification of Bacteria and Yeast Directly from Positive Blood Culture Broths
Christine Ginocchio
North Shore-LIJ Health System Laboratories
Long Abstract | Bio | Financial Disclosure
We evaluated VITEK MS (IVD Knowledgebase v2.0 and Saramis RUO database v4.10) for detection of bacteria (28 genus, 61 species) and yeast (4 genus, 7 species) directly from 632 blood culture broths representing 407 bacteremic episodes. MS-IVD identified more organisms (84.7%) than MS-RUO (80.1%), and combined use of MS-IVD and MS-RUO increased identification to 86.4%. Correlation between IVD/RUO-MS (bloods) and conventional testing (isolates) was 99.4% at genus-level and/or species-level. MS-IVD and MS-RUO results (isolates) correlated 95.8% to genus-level and/or species-level. MS-IVD/RUO results (isolates) correlated 97.9% with conventional testing and MS-IVD/RUO correctly identified 4 isolates, misidentified to genus by conventional testing.
Applications of Metabolomics in Biomedical Research
Caroline Johnson
Scripps Research Institute
No Long Abstract
In this course the utility of metabolomics will be discussed and its application to different areas of biomedical research. One of the major uses of metabolomics is biomarker discovery, which can help determine mechanisms of diseases; here their ultimate value in biomedical research will be discussed. Another topic introduced will be drug metabolism studies, which have heavily benefited from metabolomic analysis. Numerous novel metabolites and toxic modes of action have been discovered, and the emerging field of pharmacometabolomics can aid in predicting responders and non-responders. The course will end with a discussion on the integration of other –omic technologies with metabolomics, such as the rising field of metagenomics for correlating bacterial-conferred metabolic dysregulation with disease. This course is meant to introduce the non-expert to potential applications of 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
Development of Reference Measurement Systems
Chair: Don Mason
Track 2
Harbor Ballroom 2
Small Molecule Analytes - Dried Spot Analysis
Chair: Brian Rappold
Track 3
Harbor Ballroom 3
New Advances
Chair: Randy Nelson
Track 4
Marina 6
Basics: Proteomics
Chair: Cory Bystrom
10:45 AM
11:10 AM
The Development and Utilization of Liquid Chromatograph-Tandem Mass Spectrometry Methods for Value Assignment of Vitamers in Clinical Standard Reference Materials®
Johanna Camara
National Institute of Standards and Technology
Long Abstract | Bio
The National Institute of Standards and Technology (NIST) has utilized isotope-dilution liquid chromatography tandem mass spectrometry to value assign vitamin D, folate, and vitamin B6 metabolites in a variety of clinical Standard Reference Materials (SRMs). NIST currently offers and is developing SRMs which include single level materials, as well as materials specifically focused on classes of related vitamers spanning multiple levels and encompassing value ranges relevant to the clinical measurement community. The availability of new standard materials, as well as advances in separations and mass spectrometry, allow NIST to continually expand the number of value assigned vitamers in clinical SRMs.
Direct Analysis of Dried Blood and Urine Samples by PaperSpray-MS: Applications in Pain Management, Compliance, and Therapeutic Drug Monitoring
Nicholas Manicke
Indiana University-Purdue University Indianapolis
Long Abstract | Bio
PaperSpray is a rapid method for the direct analysis of dried matrix spots by MS. Analysis is performed by depositing the sample on a porous substrate (e.g. paper) that has been cut to a sharp point followed by simultaneous extraction and ionization directly into the MS. Two applications will be discussed. 1) Screening of illicit drugs and pain medications from dried urine spots on an orbitrap MS. 2) Quantitative analysis of tacrolimus and other immunosuppressive drugs from dried blood spots with LLOQs as low as 1.5 ng/mL on a triple quad. Cross-validation data with HPLC-MS/MS assays will also be presented.
Protein Biomarkers Validation Platform
Dobrin Nedelkov
Biodesign Institute, Arizona State University
Long Abstract
Described in this presentation is a Protein Biomarkers Validation Platform for performing MS-based assays at a scale, throughput, and cost that justifies their use in biomarker validation and clinical laboratories. The platform is comprised of four key modules: 1) Sample preparation, 2) Immunoaffinity isolation and protein elution, 3) Mass Spectrometry analysis, and 4) Data processing. The platform can tackle the high-demanding task of analyzing thousands of samples per day, with analytical performances and cost basis matching those of existing enzymatic immunoassays. At the same time, the platform can detect and quantify protein variants, which could have yet-undiscovered pathophysiological implications and potential clinical utility for specific diseases. The platform's performance is demonstrated through analysis of insulin-like growth factor 1 (IGF1) from >1,000 human plasma samples.
De novo Biomarker Development: Learning the Ropes: Part 1
Jennifer Van Eyk
Cedar Sinai Medical Center
No Long Abstract | Bio | Financial Disclosure
There is a need to develop new biomarkers to provide quantitative and actionable information essential for good clinical decisions. This information can aid in screening, prognosis, diagnosis and for therapeutic monitoring or improving the cost effectiveness of providing healthcare. The technologies involved in developing new markers are generic but unique characteristics of the targeted clinical area impacts the development strategy. Overall, the biomarker development process consists of multiple steps, each increasing the probability of producing clinically useful assays. This session provides insight into the strategies for discovery to increase the probability of ultimate success and outline traditional and emerging approaches for validation to hone the panel. Finally, an expert panel will discuss the challenges and obstacles in the transition of new markers to clinical practice.
11:10 AM
11:35 AM
Critical Considerations for the Use of Isotopically Labeled Protein Standards for SI-traceable Quantification in Serum: Human Growth Hormone as a Model System
Caroline Pritchard
LGC
Long Abstract | Bio
To manage and inform diagnostic or therapeutic decisions, measurement results which are accurate, specific and comparable between laboratories are required. Two challenges associated with this are the definition of the measurand and the commutability of the reference standard used. Once the measurand is defined, the next step in improving standardization is developing traceable quantification methods for proteins in biological fluids. A reference method for the quantification of human growth hormone (rhGH) in serum has been developed using multi-step sample clean-up, tryptic digestion, and isotope dilution mass spectrometry. Critical considerations for using isotopically labeled rhGH as the internal standard are described.
Overcoming All Issues Associated with Dried Blood Sampling? the Case of CYP1A2 Phenotyping
Christophe Stove
Laboratory of Toxicology, Ghent University
Long Abstract | Bio
The standard approach for CYP1A2 phenotyping is the determination of the paraxanthine/caffeine ratio in plasma, at a fixed time point after intake of the CYP1A2 substrate caffeine. Here we set up an LC-MS/MS method for DBS-based CYP1A2 phenotyping, with special attention for methodological issues associated with DBS sampling. Apart from classical validation parameters, we evaluated the (combined) impact of hematocrit, volume spotted and punch location and we compared capillary DBS concentrations (non-volumetric application, direct from the fingertip) with those in venous blood, DBS and plasma in >70 volunteers. While several of the evaluated parameters had a significant impact on the concentration of the individual compounds, the paraxanthine/caffeine CYP1A2 phenotyping ratio essentially remained unaffected. These findings suggest that CYP1A2 phenotyping is an ideal DBS application.
Towards the Development of a µHuman: Dynamic Metabolic Response Analysis of 3-Dimensional Hepatocyte Bioreactor to Acetaminophen Exposure Using UPLC-IM-MS
Cody Goodwin
Vanderbilt University
Long Abstract | Bio
An integral aspect of the integration of multiple organ-mimics for streamlined drug testing is the determination of methods for bioreactor viability, baseline, and condition interrogation in a non-destructive manner. Herein, we describe the analysis of effluent from a three-dimensional liver bioreactor using ultraperformance liquid chromatography-ion mobility-mass spectrometry and multivariate statistical analysis and self-organizing map-based approaches to data distillation. Bioreactor responses to acetaminophen are described, and metabolic perturbations are correlated to physiological reasoning and proposed metabolic pathway disruption. These analyses demonstrate an untargeted approach for benchmarking organ-mimic metabolic profiles and uncovering biological ramification of xenobiotic stimuli.
Biomarker Development: Learning the Ropes: Validation: Part 2
Dawn Chen
Quest Diagnostics Nichols Institute
No Long Abstract | Bio | Financial Disclosure
Validation of candidate biomarkers requires moving to a quantitative assay able to assess increasingly large number of samples. Traditionally, this has involved ELISA assays, which uses antibodies to different epitopes but which can be influenced (low sensitivity/specificity) by a number of factors including cross reactivity and interferences from the matrix or analyte. Recent developments in protein-based mass spectrometry methods with respect to sample preparation and instrumentation allows for absolute quantification of an analyte based on accurate measurement of representative and unique peptides. These targeted MS methods, called multiplex or selective reaction monitoring (MRM or SRM, respectively) can produce equivalent data to an ELISA. MRM assays have additional advantages with respect to multiplexing and quantification of disease-induced post-translational modifications.
11:35 AM
12:00 PM
Development of a Reference Measurement System for Urinary 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 urinary albumin is vital to clinical diagnosis. Therefore to address urinary albumin measurement precision, we have developed a reference measurement procedure that utilizes isotope dilution-mass spectrometry (ID-MS) and multiple reaction monitoring (MRM) to quantify urinary albumin and a primary reference material to be used as a calibrator for higher-order urinary albumin methods. The MS-based quantitation of urinary albumin provides both accurate and repeatable measurements at both micro- and normoalbuminuria levels, which can facilitate early diagnosis of kidney dysfunction.
Recent Developments in Automated Dried Plasma Spot Analysis Using a Novel Membrane Size Exclusion Card
Jack Henion
Quintiles Bioanalytical and ADME Labs
Long Abstract | Bio | Financial Disclosure
Recent advancements in LC/MS technologies have revived interest in using dried blood spots (DBS) in routine bioanalysis. This approach is useful for patients who are challenged with conventional larger volume blood collections. To circumvent possible issues such as hematocrit and compound instability a novel two-layered membrane-cellulose substrate device is described that enables the collection of dried plasma spots (DPS). The new device is suitable for applications of micro samples of whole blood (5 to 25 µL). In order to construct an effective device, materials from several major polymer membrane suppliers as well as high-quality scientific paper suppliers were acquired and tested. The preferred membrane and paper materials were then constructed into standard dimension ‘cards’ which are amenable to modern automated bioanalyses using on-line LC/MS/MS.
Inertial Imaging of Individual Biomolecules with Nanomechanical Systems
Scott Kelber
California Institute of Technology
Long Abstract | Bio

Selim Hanay will speak in place of Scott.

The spatial distribution of mass within an individual analyte can be imaged - in real time and with molecular-scale resolution - when it adsorbs onto a nanomechanical resonator. Each single-molecule adsorption event induces discrete, time-correlated perturbations to the modal frequencies of the device. We show that by continuous monitoring of multiple vibrational modes, the spatial moments of mass distribution can be deduced for individual analytes, one-by-one, as they adsorb. We validate this new method for inertial imaging using both experimental multimode frequency-shift data and finite-element simulations - to analyze the inertial mass, position-of-adsorption, and the shape of individual analytes. The spatial resolution is limited only by frequency noise and can achieve atomic scale with advanced NEMS devices.
Expert Panel Discussion: Learning the Ropes: Part 3
Ian Wright
Advanced Clinical Biosystems Institute
Long Abstract | Bio | Financial Disclosure
There is a need to develop new biomarkers to provide quantitative and actionable information. The technical ability and strategic approach on how to move from discovery through validation has been developing over recent years. Mass spectrometry has been central to this. The next stage is to move these new assays into clinical realm. This session will be a panel discussion with Shijun (Simon) Sheng (Senior application scientist, Thermo), Cory Bystrom (Director, Research and Development, Cleveland Heart Lab) and Ian Wright (ACBI ). Their experience with IVD and MS manufacturers as well as their interaction with academic and clinical labs will provide insight into the various aspects involved. They will highlight the challenges and successes of using MS- based methods for accurate quantification. A vision for the diagnostic industry and MRM assays will be discussed.
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)

Sigma-Aldrich
Harbor Ballroom 1

Leveraging Sample Preparation and Chromatography to Demystify LC/MS Sample Analysis
Craig R. Aurand, Senior Applications Scientist, Bioanalytical Research, Supelco
David S. Bell, R&D Manager, Pharmaceutical & Bioanalytical Research, Supelco

As LC/MS becomes more widely accepted within the clinical setting, more emphasis is being placed on the rapid generation of data to enable timely assessment of patient conditions. With the need for rapid sample turn-around, there is risk of sacrificing quality for throughput. The power of the "upfront" aspect of the LC/MS workflow – the sample preparation and chromatographic separation – is often poorly understood and often overlooked as a means to increase throughput and improve data quality. This seminar will expose users to easy-to-implement sample prep and chromatographic tools, while demonstrating strategies to help demystify the sample prep process. Discussions on leveraging chromatographic selectivity along with sample prep techniques for increasing throughput and accuracy will be covered. Attendees will come away with strategies to improve LC/MS analyses in terms of speed, sensitivity, accuracy and precision.

Thermo Scientific
Harbor Ballroom 2

Transforming Approaches to Clinical Research and Forensic Toxicology Screening - Employing Ultra Fast Triple Quad and HRAM Orbitrap Technology with Novel Ionization Technology
Marta Kozak, Applications Manager, Thermo Fisher Scientific; Kristine Van Natta, Applications Specialist, Thermo Fisher Scientific

Pre-Register

In this workshop two speakers will describe how they utilized the latest LC/MS technologies to perform ultra fast screening. Marta Kozak, applications manager, and Kristine Van Natta, applications specialist, will describe how they used the latest ultra fast QQQ technology and HRAM technology for screening panels of analytes with fast HPLC, Laser Diode Thermal Desorption, and Paperspray. This workshop will feature data presentations and interactive discussion on applying novel technologies to fast, quantitative screening to a variety of clinical research and forensic toxicology analytes.

Biotage
Harbor Ballroom 3

Advances in Sample Preparation for Mass Spectrometry Assays in Clinical Toxicology
Stephanie J. Marin, Ph.D., R&D Investigator II, ARUP Institute for Clinical and Experimental Pathology, ARUP Laboratories, Inc.

Developing a new method requires knowledge of sample prep options and an understanding of extraction mechanisms. This talk will show results from SPE and SLE ( Supported Liquid Extraction) methods which have been evaluated for TOF assays, while developing a single sample prep protocol for about 70 compounds. Ms. Marin will then discuss why they decided to move forward with the ISOLUTE SLE+ to do these extractions, and show the method development for both cord and urine assays.

IONICS Mass Spectrometry
Marina 6

Better diagnosis and treatment of thyroid diseases using sensitive LC/MS/MS measurement of FT4/FT3
Dr Steven J Soldin PhD, FACB, FCACB; Senior Scientist, National Institutes of Health, Bethesda, MD, USA

Pre-Register

In this workshop Dr. Steven Soldin from the National Institutes of Health (NIH) will discuss problems with current immunoassays to measure T4, T3, FT4 and FT3. The commonly used direct analogue immunoassays for the measurement of FT4 have been shown to have poor performance at the upper and lower limits of the FT4 reference interval. Also, recent advances in testing for thyroid illness and the role of mass spectrometry in improving the diagnosis and treatment of hyper and hypothyroidism will be addressed.

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
Protein Biomarkers of Cancer
Chair: Mark Duncan
Track 2
Harbor Ballroom 2
Trends in Toxicology: High Resolution Mass Spectrometry
Chair: Fred Strathmann
Track 3
Harbor Ballroom 3
Metabolomics III: Untargeted Profiling
Chair: Jessica Lloyd
Track 4
Marina 6
Basics: Microbiology
Chair: Carey-Ann Burnham
2:30 PM
2:55 PM
Taking Protein-targeted Multiplexed Multiple Reaction Monitoring Mass Spectrometry to the Clinic
Stephen Hunsucker
Integrated Diagnostics
Long Abstract | Bio | Financial Disclosure
There is tremendous potential for MRM MS analysis of proteins in clinical diagnostics. Progress has been limited by inability to translate protein biomarkers from discovery through validation required for clinical implementation. We present successful translation and validation of a multiplexed protein MRM assay available through a CLIA-certified laboratory. The assay identifies benign pulmonary nodules with high negative predictive value to avoid unnecessary invasive procedures. Importantly, the assay provides molecular information independent of clinical data currently used to assess risk of malignancy. The assay proteins are associated with cancer-related pathways, including cell growth and proliferation, lung inflammation, and oxidative stress response.
Detection of Allenic Norleucine, a Nephrotoxic Amino Acid from Amanita Smithiana Mushrooms
Ian Garber
University of British Columbia
Long Abstract | Bio
Amanita smithiana is a poisonous mushroom, growing along the Pacific coast of North America, which is sometimes confused with the edible and highly valuable Pine mushroom. It causes acute renal failure, usually requiring dialysis. We present one such case of misidentification from British Columbia. Up until now, laboratory confirmation of A. smithiana in poisoning cases has relied on thin-layer chromatography. The chemical identity of the characteristic spot seen on TLC has been uncertain. We used LC-MS/MS to confirm that the toxin spot consists of a modified amino acid, allenic norleucine. Chlorocrotylglycine, an additional amino acid unique to this species, was also detected by LC-MS/MS from an adjacent TLC spot. Mass spectrometric detection of these two compounds should now be able to replace the more labour intensive and subjective thin-layer chromatography assay.
Untargeted Metabolomics Identifies Numerous Solutes that Accumulate when the Kidneys Fail
Pavel Aronov
Thermo Scientific
Long Abstract | Financial Disclosure
Solutes normally cleared by the kidneys accumulate and cause illness when the kidneys fail. Untargeted metabolomics is an ideal way to expand our knowledge of these "uremic" solutes. The current study employed the Orbitrap based mass spectrometer (Q Exactive MS) to profile solutes that accumulate in hemodialysis samples. The discovery phase revealed 400 uremic components in negative ESI mode and 175 uremic components in positive ESI mode. Chemical identification using MS/MS scans and accurate mass information has so far revealed novel uremic solutes including N-glutamyl heptenoic and N-glutamyl octenoic acid.
MALDI-TOF 101 for Microorganism Identification
Morgan A. Pence
Washington University in St. Louis
No Long Abstract
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an emerging diagnostic tool in the clinical microbiology laboratory. MALDI-TOF MS provides identification of microorganisms within minutes, decreasing turnaround time compared to conventional phenotypic methods, and has been shown to be reliable and cost effective. The method generates a spectrum, or “fingerprint,” of ribosomal proteins, which is compared against a reference database to assign an identification and confidence value to the organism. This session will focus on the basics of MALDI-TOF MS in the clinical microbiology laboratory and discuss the two platforms that currently exist, the Bruker Biotyper and bioMérieux’s VITEK MS.
2:55 PM
3:20 PM
High-Resolution Mass Spectrometric Quantification of Peptides in Clinical Samples
Bruno Domon
Luxembourg Clinical Proteomics Center
Long Abstract | Financial Disclosure
New hybrid mass spectrometers with high resolution and accurate mass capabilities have opened new avenues in quantitative proteomics. Targeted clinical analyses, routinely performed on triple quadrupole instruments in selected reaction monitoring mode, were replicated on a high-resolution quadrupole-orbitrap instrument operated in parallel reaction monitoring (PRM) mode. The trapping capability is well indicated to isolate and analyze peptides in tiny amounts and thus dramatically increased the dynamic range while providing highly selective measurements. The PRM technique was applied to the evaluation of large panels of putative makers in clinical plasma samples and showed a gain in selectivity and an increase in the confidence of measurements in complex matrices. The PRM analysis of lung cancer candidate markers resulted in a clear discrimination of the disease stages and subtypes.
High Resolution Mass Spectrometry: The Comprehensive Toxicology Screen of the Future?
Jennifer Colby
University of California San Francisco
Long Abstract | Bio
High resolution mass spectrometry (HRMS), which allows exact determination of molecular weight, has been proposed as an alternate to immunoassay screening of biological samples. One advantage of HRMS is the ability to screen for unknown compounds. The molecular formula of the target ion is predicted from the measured mass, then the user must determine which formula and structural isomer best fit the data. In our laboratory the search process has been empirical, often requiring multiple iterations. We present here a strategy for optimizing the parameters required for untargeted searches as well as final parameters and method performance on blinded samples.
Characterization of Metabolomic Profiles of Nipple Aspirate Fluid Using UPLC-QToF; Potential Insights for Novel Breast Cancer Risk Biomarkers
Jessica Miller
University of Arizona Cancer Center
Long Abstract | Bio
Nipple aspirate fluid (NAF) components are constantly secreted, metabolized and re-absorbed by the epithelial lining of the breast duct making NAF a viable biologic to mine for novel early detection and cancer surrogate biomarkers. We characterized metabolomic profiles of NAF and plasma from healthy pre (N=10) and postmenopausal (N=19) women using UPLC-QToF. NAF was composed of lipids, drug metabolites and food additives as well as multiple bioactive food components. Plasma primarily contained endogenous metabolites. This work suggests NAF contains novel biomarkers distinct from plasma; further profile characterization, metabolite identification, and determination of potential breast cancer relationship are ongoing.
MALDI-ToF-MS for Mycobacteria & Mold
Susan Butler-Wu
University of Washington
No Long Abstract | Bio | Financial Disclosure
The use of MALDI-ToF-MS for the identification of routine bacterial isolates in the clinical microbiology laboratory is becoming increasingly commonplace. Similarly, there is a large body of published literature describing the performance of this method for these organisms. However, the application of MALDI-ToF-MS to mycobacteria and mold species poses unique challenges such as the need for initial extraction, safety considerations and database composition. In this session, we will review published extraction methods as well as the performance of both commercial MALDI-ToF-MS systems for these organisms. The practicalities of implementing MALDI-ToF-MS for mycobacteria and mold identification will also be discussed.
3:20 PM
3:45 PM
Analysis of Serum Glycopeptides from Aggressive and Non-aggressive Prostate Cancer Patients Using Automated Glycopeptide Extraction and Parallel Reaction Monitor
Robert Harlan
Johns Hopkins University
Long Abstract | Bio
Current prostate cancer screening techniques cannot distinguish between aggressive and non-aggressive forms of tumors. In this study, we used an automated solid phase extraction (SPEG) of glycopeptides from serum samples of men with varying degrees of prostate pathology and analyzed with parallel reaction monitoring (PRM) in an attempt to discover prostate cancer biomarkers. Seventy-five serum samples were obtained representing patients with non-cancerous, aggressive, or non-aggressive prostate cancer. Glycopeptides were isolated from 40 µl of serum from each patient via automated glycopeptide capture using liquid handling robotic system (Versette, Thermo Fisher Scientific). Samples were then analyzed using a Q Exactive mass spectrometer by PRM assay (Thermo Scientific). We identified six potential serum biomarkers that that may distinguish aggressive from non-aggressive forms of prostate cancer. In addition, we identified three potential serum biomarkers that may be used to distinguish between cancer and non-cancer.
All Ions MS/MS in TOF/MS: A Reliable and Cost-effective Alternative to Collecting Accurate Mass Data on Molecular and Fragment Ions from QTOF/MS
Roy Gerona
UCSF
Long Abstract | Bio | Financial Disclosure
All Ions MS/MS in TOF/MS is a technique that uses multiple fragmentor voltages to induce in-source fragmentation allowing simultaneous generation of molecular and fragment ions in full scan MS mode. Cycling of fragmentor voltages during each time segment in a TOF/MS run from low to high energy allows formation of molecular and fragment ions, respectively. We successfully demonstrated the ability of All Ions MS/MS in an Agilent LC 1200- TOF/MS 6230 to generate and identify fragment ions of 75 designer drugs in a pyschostimulants drug panel. This allowed for significantly improving the accuracy of drug identification by LC-TOF/MS including discrimination between structural isomers. .
The Cancer Isotopolome in Whole Cells by Metabolomics
Ying-Jr Chen
Washington University in St. Louis
Long Abstract | Bio
NMR and mass spectrometry (MS) are powerful analytical technologies that provide complementary information about the cancer metabolome and biomass profile. Here we describe software for integrating MS and NMR data that filters MS-based metabolomic features which are not identified by NMR. By applying this approach to cancer cells grown on specific isotope labels, we are able to reduce the thousands of peaks detected in metabolic extracts by MS to a small number that are consistent with the analytes observed in whole-cell NMR. The analysis of whole cells by NMR also allows for quantitative assessment of the amount of label that ends up in protein, DNA, and lipid membrane biomass. Comparing quiescent cells to cancer cells, our approach has revealed that the metabolic reprogramming observed in cancer results in major alterations in the precursors used for biomass production.
Advanced Applications for MALDI-TOF MS in Clinical Microbiology
Christopher Doern
Virginia Commonwealth University Medical Center
No Long Abstract | Bio
This educational session will focus on advanced applications of matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) as they pertain to the diagnosis of infectious diseases. Three primary topics will be discussed: 1) the use of MALDI-TOF MS for the detection of beta-lactamases and other resistance determinants, 2) phylogenetic typing of bacterial organisms, 3) direct identification of organisms from positive blood cultures and patient specimen. The educational session will focus on describing the methods used in these techniques as well as reviewing the literature that addresses their performance.
3:45 PM
7:00 PM
HOSPITALITY
@ Shoreline Patio

Enjoy the San Diego evening down by the Marina with heaters and fire pits. Food & Drinks provided. At Shoreline Patio by the pool.
7:00 PMCONFERENCE CLOSED