Description:

This training session will provide attendees with actionable best practices for optimizing chromatography and addressing common troubleshooting challenges. Using real-world examples, participants will explore advantages as well as common pitfalls when optimizing autosampler settings, LC pump configurations, column chemistries, mobile phase compositions, and gradients. The session will also demonstrate how these techniques can be effectively applied to identify and resolve common issues with sample injection and chromatographic separations.

Pearls:

After attending this training, participants will be able to:
1. Implement best practices for optimizing chromatographic conditions to improve performance and data quality
2. Design efficient experiments to determine best autosampler settings, columns, mobile phases, and gradients for specific applications
3. Utilize troubleshooting techniques demonstrated in the session to identify and resolve common sample injection and chromatography issues in their laboratories.

This practical training focuses on the real-world implementation of LC-MS/MS kits in routine diagnostic laboratories. The course will cover critical pre-purchase questions to ask vendors, including system compatibility, existing users, validation status, and available (often unpublished) knowledge. Stop/Go decision points are addressed, helping laboratories evaluate implementation challenges, decide when additional vendor support is justified, and recognize when transitioning to an LDT approach may be more appropriate. The training also covers common troubleshooting challenges, such as limited transparency around mobile phases and columns, and discusses strategic use of kit components such as calibrators and quality controls. Using successful implementation examples, participants will learn which validation/verification experiments are required and how to apply practical tips and shortcuts beyond the kit instructions.

Take-home pearls:
By the end of this practical training, participants will be able to:
1) Ask the right vendor questions before committing to a kit and during implementation of the kit.
2) Identify Stop&Go points during implementation of the kit.
3) Select kit components flexibly and effectively.
4) Define which validation and verification experiments are essential.

This session will explore techniques used to ensure analyte specificity in LC-MS analyses for small molecules. The session will apply to both nominal mass as well as high-resolution instruments and will include preanalytical considerations, chromatography considerations, use of appropriate internal standards, and analyte match criteria. Method validation procedures to evaluate potentially interfering compounds and ion cross talk will be discussed. The effects of optimizing ionization energy on fragmentation patterns, and the use of accurate mass and isotopic patterns will be examined. Result reporting and interpretive comments for suspicious but unconfirmed compounds will be addressed. Audience participation will help foster the learning experience.

Pearls:
After this session participants will be able to:
1) Relate factors that can interfere with MS assays.
2) Identify sources of ion cross talk.
3) Describe effects of ionization energy on analyte fragmentation.
4) List analyte match criteria to improve assay specificity.

Is perfect precision possible? This practical training session makes the case for sample preparation as the chief determinant of controlling assay precision and getting as close as possible. We’ll walk through key fundamental practices and experiments to maximize the analytical control of your assay. During the session we’ll cover pipetting strategies, internal standards, dilution strategies, carryover control, and ruggedness by design. Throughout the session we’ll use brief clinical case vignettes to show how “prep first” thinking increases the quality of your clinical batches, therefore reducing sample re-extractions and maintaining proper turn around time. Lastly, we’ll briefly compare standard sample cleanup choices such as protein precipitation, phospholipid removal, and SPE through the lenses of measurement requirements, turnaround time, automation, safety, and cost.
This session is not an all-encompassing guide to sample preparation; rather, it equips you with a practical toolkit to design, execute, and interpret the highest-impact experiments first.

Take home “Pearls”
1) Attendees will learn key experiments design assays with tight analytical control.
2) Contextualize real-life examples to key experiments discussed.
3) Compare/contrast common extraction prep schemes and balancing measurement requirements with workflow needs like speed, automation compatibility, safety, and cost.

To provide best practices for development, validation and implementation of MS methods for the quantitation of therapeutic monoclonal antibodies (tmAbs). The goal of this session is not to provide method specific details from our published methods, but instead to share the lessons learned as we streamlined our pipeline for TDM for new tmAbs. Multiple methods have been utilized for the enrichment of tmAbs to include enriching either all immunoglobulins, a class or subclass, or for a specific antibody. Automation efficiencies has been implemented at various stages. Detection has included either peptide or light chain from the tmAb of interest. Instrumentation has spanned MALDI for screening to single or multiplex HPLC systems connected to either a triple quad, orbitrap or time of flight MS for quantitation. The speakers will share some of the headaches, best practices and innovations of being at the forefront of a new analyte, extraction method or platform.

Take Home Pearls:

1. Describe best practices that improve assay accuracy and precision, including matrix selection, internal standard usage, and selection of appropriate reference materials.
2. Explain how antibody type (chimeric, humanized, subclass specific) influences method selection for clinical therapeutic drug monitoring.
3. Identify key factors that determine appropriate enrichment and detection strategies for quantitating therapeutic monoclonal antibodies using mass spectrometry.
4. Differentiate the advantages and limitations of peptide based versus intact light chain mass spectrometric detection across available instrument platforms (triple quad, orbitrap, TOF, MALDI).
5. Outline a streamlined development pathway for implementing new monoclonal antibody quantitation assays in a clinical laboratory setting.

Planned publication of a revision of C-62: Liquid Chromatography-Mass Spectrometry Methods by CLSI is set for August of 2026. The third edition of this document includes significant revisions and additions to the best practice guidance for LC-MS in the medical laboratory. This workshop will initiate discussion on the updates, providing attendees with up-to-date information and background on the new recommendations with members of the C62 document committee.

Take home pearls: Attendees will be given constructive information addressing the following features included in the C62-A3 revision:

1.) Updated performance metrics (Signal-to-noise/LoD/LoB/LLMI/ULMI).
2.) Clarification of LC-MS specific constraints to CLSI Evaluation Protocols (EP docs).
3.) Detailed Applications of Ion Ratios (e.g. different modes of acceptable range generation).
4.) Processes of verification of critical reagents.
5.) Quality controls for multiplexed analysis.