Day 3

Training Program

Learner Description

The training program is an advanced LC/MS workflow program that assumes the trainee matches the following profile: Possesses a good understanding of Customer Familiarization and deliveryHas attended any LC courseHas attended Blueline or Legacy classPossesses Good Laboratory PracticesUnderstands Lab safetyLearner ResponsibilitiesCompletes assigned training tasksCompleted Customer Familiarization both training and field implementationDevelops an understanding of LC/MS workflowUnderstands how to use analysis software and IDA

Sample Prep.(types and examples) Mass Spec Consideration (Triple/Traps vs TOF) Triple/Traps analyzer Modes Selecting analyzer mode for quantitation LC Method Considerations  (Column/Mobile Phase/buffer)

Prerequisites: Completion of an LC Training classCompletion of a Blueline or legacy trainingFor those with electronic background -Completion of the Fundamentals of LC/MS  program Pre-Learning (training modules)ELearning on MyDevelopmentInstructor Led Training (ILT)Instruments covered HPLC: Shimadzu or AgilentMass spectrometer: Blueline or legacySafety (EHS Training) Personal safety and the use of PPE (Personal protective equipment)Nitrile powder-free glovesLab coatSafety glassesElectrical safety Good laboratory practices Lab safety and safety of others Complying with Customer site requirements Proper Pipetting Proper measure weights Good document keeping Sample Preparation Types of Sample preparation (Liquid/liquid Extraction, Protein Precipitation, Solid Phase Extraction)Results of poor sample prepResults of good sample prep Basic Troubleshooting Tubing connectionsSoftware (Analyst, Multiquant and IDA)Sample PreparationChromatographyLC/MS Method Development Concepts Types of  compounds to separateQualitative vs Quantitative analysisLC/MS system considerationTypes of Columns to useMobile phase & modifier selectionLC system selection Gradient vs isocratic selection

Compound Optimization High Flow Optimization Source & gas Optimization LC/MS Operation Aquisition/batch

Applied LC/MS

Quantitation/Data Generation IDA MRM_IDA and library searching Data processing/Review Quantitation/MultiQuant

Day 4

Practise/Review Exercise 1

Program Description

Day 5

Practice/Review Exercise 2

Fundamentals of LC/MS Workflow

The Applied LCMS training course covers the major areas a Triple Quad / QTRAP customer will use. This will assist the engineer to better understand the many non-hardware related aspects of the covered systems. The course includes sample preparation, LC method modifications, compound optimization, quantitation including MultiQuant and IDA. Included in the training is extensive practice and advanced use of Analyst software. The course is taught from a Customer’ use perspective that highlights the potential areas of issues that may typically be encountered along the LC/MS workflow. This will allow the service engineer to gain experience on the workflow and to better diagnose potential application issues. The starting point of the course is a working system. Throughout the course, different scenarios will be covered that represent real-life examples seen in the field. Out of scope tasksIt is important to understand that the Field Service Engineer will NOT be taught the following items:Servicing any instrumentsLearning/using all related software packages for Triple Quad / QTRAP

LC-MS/MS Operation Preparation Guideline Mobile phasesWash Solvent for the Autosampler and Syringe CleaningTriazine DilutionsInfusion Dilution for OptimizationSystem ConfigurationOptimize for the Compound (Manual and Automated Compound OptimizationIon Production & High Flow Optimization Source/Gas ParametersPlumb the HPLC System and Load the SamplesInformation Dependent Acquisition Brief overviewMRM_IDALibrary searchingAcquisitionBuild the Acquisition MethodCreate the Acquisition BatchSubmit the BatchReview Acquired Data in Explore ModeData AnalysisExplore modeAnalyst® QuantitationMultiQuant™ software Data ProcessingBuilding a Quantitation MethodCreate a results TableBatch creation including QuickQuanStandard curve generationReview of ResultsTroubleshooting LC/MS  General over view of troubleshooting processUnderstanding these concepts to distinguish between an application vs hardware issueParticipantsMaximum 6 Service Technicians (if 3 instruments available)

Day 1

Fundamentals LC/MS Workflow *Performing Compound and source/gas Optimization  Understanding and using LC Method variables  Demonstrating basic proficiancy in sMRM  How to set up a basic MRM IDA method  General competancy using Analyst software including select scripts  Demonstrate proficiency using Analyst Explore  Demonstrate proficiency using Multiquant and analyst quantitation  Demonstrate proficiency using Analyst Quantitation  Sample Preparation theory  Performing Sample Preparation  * In workflow since this is not related to instrument tuning and optimization  General Laboratory Practices  4.1 calculations  4.2 Weighing, measuring and pipetting  4.3 Safety

Day 2

Agenda

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The evaluation for the associate’s training completion will be based on the following competencies:

Competency

This course will cover the operational theory and workflow of LC/MS. Topics that bring familiarization to the customer workflow (such as method development, source and compound optimization, method creation and data analysis) will be covered in detail.  The learner will acquire the knowledge, tools and skills to successfully run through the LC/MS (customer) workflow. The FSEs will learn how to develop and optimize an LC/MS method. The learner will also acquire knowledge to quantitate data using both Analyst Quant and Multiquant. Out of scope tasksIt is important to understand that the Field Service Engineer will NOT be taught the following items:Servicing any instruments

The training program will assume the trainee matches the following profile: An FSE with an electrical backgroundHas attended any LC courseHas attended Blueline or Legacy classHas Good Laboratory PracticesUnderstands Lab safetyLearner ResponsibilitiesCompletes assigned training tasksDevelops an understanding of LC/MS workflow

Prerequisites: Completion of an LC Training classCompletion of a Blueline or legacy trainingPre-Learning (training modules)ELearning on MyDevelopmentInstructor Led Training (ILT)Instruments covered LCShimadzu or AgilentMass spectrometerBlueline or legacySafety (EHS Training) Personal safety and the use of PPE (Personal protective equipment)Nitrile powder-free glovesLab coatSafety glassesElectrical safetyGood laboratory practices Lab safety and safety of othersComplying with Customer site requirementsCustomer Familiarization Documentation Review LC-MS/MS Operation Preparation Guideline Mobile phasesWash Solvent for the Autosampler and Syringe CleaningTriazine DilutionsInfusion Dilution for OptimizationSystem ConfigurationConfigure the System for InfusionOptimize for the CompoundPlumb the HPLC System and Load the SamplesAcquisitionBuild the Acquisition MethodCreate the Acquisition BatchSubmit the BatchReview Acquired Data in Explore ModeTroubleshooting LC/MS  General over view of troubleshooting processUnderstanding these concepts to distinguish between an application vs hardware issueParticipantsMaximum 6 Service Technicians (if 3 instruments available)

Fundamentals LC/MS Workflow:Understands how to prepare standard samples and HPLC mobile phaseCan set up an LC/MS systemCan perform Compound Optimization on the Mass SpecCan create an LC/MS methodCan create a sample batchCan process data to obtain calibration curves and results from MultiQuantDistinguish between application and an LC/MS hardware issue

LC/MS Method Creation Building an acquisition methodCreating the acquisition BatchSubmitting a BatchReview acquired data in explore mode Building a Quantitation MethodCreate a Results table Data Analysis (Multiquant and Analyst Quant)

LC-MS overview (theory) Customer Familiarization Document overview Triazine LC/MS WorkflowMobile Phase preparationPreparation of Triazine StandardsConfiguring the LC/MS System for infusion Performing Compound OptimizationSetting up LC/MS to run samplesIon Production and High Flow Optimization

The evaluation for the associate’s training completion will be based on the following competency: