ASMS: Agilent LC/TQ user meeting

LC/TQ user meeting

• Location: Baltimore Marriott Inner Harbor, 110 S Eutaw St, Baltimore, MD 21201
• Room: Ballroom
• Date: Sunday, 1 June 2025
• Time: 8:30 am - 12:00 pm

Register

8:30 AM – 9:00 AM Registration and Breakfast

9:00 AM – 9:15 AM Agilent Quadrupole Instrumentation: the Past, Present and Future

• Shane Tichy - Associate Vice President of Research and Development, Agilent Technologies

Agilent LC/MS quadrupole instruments have revolutionized analytical chemistry by providing sensitive, precise, and reliable mass spectrometry solutions. I’ll give a quick introduction showcasing the evolution of these instruments, from their inception to their current state-of-the-art capabilities. This talk highlights key technological advancements, such as enhanced sensitivity, faster scan speeds, and improved ionization techniques, which have expanded applications across pharmaceuticals, environmental analysis, and food safety. The future of Agilent LC/MS quadrupole instruments promises further innovation, including AI-driven data analysis, miniaturization, and increased sustainability, ensuring continued impact on scientific research and industry.

9:15 AM – 9:55 AM Single-Cell Metabolomics Using CE-NanoESI-Triple Quadrupole MS

• Jonathan V. Sweedler - James R. Eiszner Family Endowed Chair in Chemistry, Department of Chemistry, University of Illinois Urbana Champaign

Single-cell metabolomics is important for understanding cell-to-cell heterogeneity and to link cell neurochemistry to function. We are unraveling the differences between brain cells, including neurons, astrocytes, and more. We couple an Agilent 7100 CE system with the Agilent 6495C triple quadrupole LC/MS using 3D-printed parts as the CE/MS interface and use the system for single cell measurements. Different identified neurons were isolated from the central nervous system of Aplysia californica and individual neurons from the dorsal root ganglion of rodents. We performed sample preparation steps in vial inserts prepared from commercially-available 10 µL pipette tips; this enables us to work with and inject from a few microliter volumes. Other changes include a nanointerface and a mechanically-tapered capillary tip for optimized detectability. Using the CE-MS system, metabolites in individual cells were characterized using multiple reaction monitoring mode with the QQQ-MS. The CE-nanoESI-QQQ MS system demonstrated detection limits in the attomole range for a range of amino acids and neurotransmitters. We characterize a range of different cells including Aplysia californica neurons, rodent neurons, and individual endocrine cells. The combination of nanovial CE-nanoESI-QQQ MS with the 3D-printed interface, auto sampler, and fast MS2 scanning enables high-throughput, high-sensitivity, and robust metabolite analysis of single cells.

9:55 AM – 10:15 AM Comprehensive Toxicology and TDM by DBS

• Kenneth Lewis - Owner, OpAns LLC

This presentation describes our HPLC/MS/MS method that quantifies hundreds of drugs from a single dried blood spot (DBS). We will also present correlation data from over 1,000 paired urine and DBS forensic samples, demonstrating the differences between the matrices for detection of substance exposure. The analysis is accomplished through optimization of method parameters using the latest generation of HPLC/MS/MS triple quadrupole instruments. The DBS and urine samples were contemporaneously observed collections from 60 days of report and community correction centers across five states. DBS samples are collected on stabilized filter paper and dried. They are punched into microtiter plates where internal standard is added, and the analytes are extracted. The analysis is performed using an Agilent Infinity III LC system and Agilent 6495D triple quadrupole LC/MS system. The analyte list starts with the ~100 typical drugs monitored in pain management and forensic testing and has been expanded to include the CDC Emerging Drug Panel, other OTC and prescription-abused drugs, plus additional fentanyl analogs and synthetic cannabinoids. DBS reporting cutoffs were set based on the blood levels reported for therapeutic use (or typical forensic cutoffs when there is no therapeutic data). Urine cutoffs were typical levels used in clinical diagnostic testing (generally lower than forensic cutoffs). Paired DBS and urine samples were collected from 1,113 donors at 60 community corrections locations (no more than 20 donors per location). The most notable differences were methamphetamine and alcohol biomarkers. There were 232 DBS methamphetamine positives compared to 107 urine positives (50 ng/mL cutoff). Ethanol exposure was determined in DBS using 16:0/18:1 phosphatidylethanol (PEth) and ethylglucuronide/ethylsulfate (ETG/ETS) in urine. There were 187 DBS PEth positives, compared to 50 urine ETG/ETS positives.

10:15 AM – 10:30 AM Break

10:30 AM – 10:50 AM Discovery Metabolomics by Using Targeted Assays and Commercial Kits

• Kevin Cho - Staff Scientist, Department of Chemistry, Washington University in St. Louis

Metabolomics provides insights into the metabolic landscape of diseases, including colorectal cancer (CRC). While untargeted metabolomics offers broad metabolic profiling, it lacks absolute metabolite quantification. This study combines targeted and untargeted metabolomics using commercial kits to identify and quantify CRC-associated metabolic changes in human serum. Serum samples from ten Stage IV CRC subjects and ten healthy controls were analyzed using the Agilent Captiva EMR-Lipid and the Biocrates MxP Quant 500 XL kit. Untargeted metabolomics profiled over 1,500 metabolites using an Agilent 6546 LC/Q-TOF, while targeted analysis quantified 454 metabolites with an Agilent 6495D LC/TQ system. Statistical analyses revealed significant metabolic distinctions, with pathway enrichment highlighting alterations in amino acid metabolism, fatty acid metabolism, and bile acid biosynthesis. Key findings included elevated cystine and reduced levels of TMAO, phosphoethanolamines, triglycerides, and bile acids in CRC samples. This integrated approach enhances our understanding of CRC-associated metabolic dysregulation and may contribute to biomarker discovery.

10:50 AM – 11:10 AM Quantitative Measurement of 2000+ Metabolites in Biofluids Using the Agilent 6495D LC/TQ System

• David S. Wishart - Distinguished University Professor, Depts. of Computing Science and Biological Sciences, University of Alberta

Metabolomics is reaching an inflection point. Either it can continue with traditional, manually- intensive, nontargeted approaches and remain a niche field of scientific research -- or it can change to become a truly quantitative, fully automated endeavor, serving as the main vehicle to advance the next generation of clinical, pharmaceutical, environmental, and forensic applications. In this presentation I will describe how it is possible to develop and implement a truly quantitative, fully-automated metabolomic assay that can routinely measure 2000+ metabolites (including metabolite sums and ratios) in less than 20 minutes on the Agilent 6495D triple quadrupole LC/MS system. In implementing this assay (called the GIGA assay) we have taken advantage of the enhanced sensitivity of the Agilent iFunnel technology, the AI-based autotuning, and the submillisecond dwell times that allowed us to rapidly collect high-quality MS/MS data on hundreds of metabolites. The improved collision cell design of the 6495D LC/TQ also allowed us to perform much more accurate quantification, even for low S/N situations. These technology advances allowed us to rapidly evolve an assay that could measure just 650 metabolites to one that could measure 2000 metabolites with just a few weeks of effort. I will describe the assay design, the assay specifications and performance assessment, and examples of its application to real metabolomics studies. I will also describe the software we have developed (called LC-AutoFit) to automatically process the MS/MS spectra collected on the 6495D LC/TQ and the low-cost robotic systems we have created to both prepare the assay kits (in a 96-well format) and to run the GIGA assay. We believe this new, fast, low-cost, and fully-automated approach to doing comprehensive, quantitative metabolite measurement will enable metabolomics to be an integral part of next-generation applications in clinical, pharmaceutical, environmental, and forensic chemistry.

11:10 AM – 11:30 AM Enabling the Advancement of Molecular Medicine

• Rob Fraser - CSO and President, Co-Founder, Molecular You Corporation

The lack of early detection and actionability costs the healthcare industry over a trillion dollars each year. Molecular You (MY) is leading the rising research and development by providing a comprehensive multi-omic analysis that potentially provide accurate, predictive insights into health risks, biological pathways at the root cause of the risk at the earliest phase of pathogenesis to enable safe, effective and personalized interventions. The platform is a two sided solution that employs advanced artificial intelligence (AI) tools and scientific expertise to a) select high value metabolites and proteins with the supporting documentation and wet lab validation to be incorporated into our multiplexed LC/MS/MS quantitation assays and b) interpret the data and potentially provide health insights in an easy to use, practical interface that removes the friction for adopting molecular medicine. This multi-omic chipset and software pairing is regularly reassessed and updated. It is also designed to incorporate other Omic assay data in collaboration with clinical research labs. MY technologies are particularly useful for measuring a wide range of interconnected molecular, microbial, and physiological changes, including those driven by pharmaceutical interventions.  Having demonstrated MY can also support the selection and development of safe and effective therapies in collaboration with pharmaceutical companies. The expanding database of longitudinal multi-omic and EHR data is designed to be analysed by researchers to generate new knowledge into the mechanisms of health and disease towards the establishment of more effective therapeutic interventions.

11:30 AM – 11:50 AM Development of a Quantitative High-Throughput Kinase Panel to Assess Degrader Selectivity

• Paul Auger - Nurix Therapeutics

Degradation selectivity is an essential criterion in targeted protein degradation (TPD) to determine which compounds progress during drug development. To assess degradation selectivity, mass spectrometry-based methods can be used to quantitatively monitor protein abundance. Multiple reaction monitoring (MRM) targeted proteomics is commonly used to detect multiple peptides—often simultaneously—within a single method, enabling quantitative readouts for a diverse set of proteins in a single assay. The ability to multiplex these measurements allows for the design of highly curated protein panels, that can quantitate collections of proteins to determine degrader selectivity. Additionally, the robust and reproducible nature of MRM triple quadrupole mass spectrometry (QqQ) makes it ideal for high-throughput screening for degrader optimization.

Nurix Therapeutics developed a quantitative kinase panel with over 45 unique proteins for degrader screening and analysis using an Agilent 6495 series QqQ LCMS. Our data demonstrates the ability to quantify kinase abundance across multiple cell matrices, an essential step in determining matrix sensitivity and on-target selectivity in TPD molecule development.

11:50 AM - 12:00 PM Application of Targeted Mass Spectrometry Imaging in the Pharmaceutical Industry

• Andrew Bowman - AbbVie, Inc.

Within the pharmaceutical industry, the prevailing drive for potent, targeted compounds has driven the need for the confirmation of drug delivery to the target. While overall mass spectrometric instrumentation has delivered ever greater sensitivity and precision, to achieve the absolute greatest sensitivity requires the use of QqQ instruments. To date, there is no commercially available, MALDI-based, targeted imaging platform. AbbVie has worked together with both MassTech and Agilent to provide a robust, rapid, and most importantly sensitive Atmospheric Pressure Matrix-Assisted Laser Desorption/Ionization Triple Quadrupole platform for the targeted analysis of known compounds of interest, providing up to 200-fold greater sensitivity in comparison to on-market broad-spectrum MALDI instruments.

12:00 PM Thank you, Steve Berger Award, lead-in to networking lunch

• Shane Tichy - Associate Vice President of Research and Development, Agilent Technologies

12:00 PM - 1:00 PM Networking Lunch

*Please indicate whether or not you will be attending in the "Personal Information" form of the registration process.