Bioanalytic Basic LC-MS/MS training

BASIC INFORMATION

• TITLE: Basic LC-MS/MS training
• CATEGORY: LC-MS/MS training
• LOCATION: Lodz, PL
• TRAINING TIME: 3 days
• NUMBER OF PARTICIPANTS: up to 5 people
• EQUIPMENT: SCIEX QTRAP & QTOF LC-MS/MS

Bioanalytic: Bioanalytic Basic LC-MS-MS training.

TRAINING DESCRIPTION

Basic LC-MS/MS training covers the theoretical basics of mass spectrometry and familiarization with the basic operation of the instrument. Basic LC-MS/MS training is primarily aimed at those who are new to LC-MS/MS equipment or who wish to become familiar with this analytical technique. During the training, the following will be presented:

• principles of developing and optimizing quantitative and qualitative methods on low (triple quadrupole, QTRAP) and high resolution (QTOF) instruments,
• basic functionality of the software,
• procedures and objectives of instrument calibration,
• principles of maintaining the cleanliness of mass spectrometer operation.

Basic LC-MS/MS training - the program

Day I

Theoretical classes:

• Introduction to LC
• Introduction to mass spectrometry (basics)
• Quantitative analysis.

Practical part:

QTRAP:

• QTRAP scanning modes on the instrument - Q1, , MS2, MRM, demonstration and discussion on the example of a selected pesticide of the fragmentation spectrum and its fragmentation pathway.
• MRM pair formation (manual, semi-automatic, automatic).
• optimization of ion source parameters -T-conector
• explanation of the main parameters related to LC (column selection, gradient vs isocrat, autosampler)
• LC-MS/MS method setup and optimization

QTOF:

• manual and automatic optimization of MRM^HR
• ion source and LC parameters (transfer from method developed above)

Sequence of quantitative analysis on both instruments.

Day II

Quantitative analysis:

• data processing and analysis,
• reporting - Sciex OS (QTRAP, QTOF).
• Discussion of types of integration algorithms (MQ4, autopeak) and integration parameters.
• independent selection of integration parameters and plotting of master curves.
• independent data analysis and generation of analysis report.

Qualitative analysis:

• Theoretical introduction - introduction to qualitative analysis:
• qwadrupole modes (Prec/NL),
• trap modes (EMS, EPI, MS3),
• QTOF modes (TOF MS, TOF MS/MS).
• mixed methods,
• targeted vs. untargeted analysis,
• DDA (IDA) - discussion of method development,

Practical part:

• creation of qualitative methods on QTRAP (Prec-IDA, MRM-IDA), other parameters (LC, ion source) as the previous day,
• LightSight - discussion of operation, creation of MRM-IDA multimethod for selected pesticide and analysis of samples (night).
• QTOF - creation of IDA method, other parameters (LC, ion source) as the previous day.

Sequence of qualitative analysis overnight.

Day III

Qualitative data analysis:

QTRAP:

• Discussion of Prec-IDA, MRM-IDA results - what we see in different types of MS and MS/MS scans. Manual review of data and via IDA Explorer (SciexOS).
• Analysis of results in LightSight, comparison with Prec-IDA, for example - advantages/disadvantages

QTOF:

• search for potentially present compounds in the sample,
• manual comparison of blank vs. test sample,
• manual by XIC extraction (by sum formula),
• Formula finder - derivation of sum formula on TOF MS data and verification of fragmentation on TOF MS/MS data
• search in IDA Explorer mode - application of filters: similarity, isotopic distribution, mass defect,
• database searching,
• Molecule Profiler - demonstration of software capabilities on selected pesticide example,

Instrument calibration:

• resolution, mass-to-charge ratio, intensity,
• Q1 and Q3, LIT (MS and MS/MS), TOF MS and TOF-MS/MS
• Automation vs. manual calibration,

LC-MS/MS application examples

Summary

Discussion