Bioanalytic Quantitative LC-MS/MS analysis training

BASIC INFORMATION

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

Bioanalytic: Bioanalytic Quantitative LC-MS-MS analysis training.

TRAINING DESCRIPTION

Quantitative LC-MS/MS analysis - the training is designed for laboratory personnel using LC-MS/MS systems for quantitative analysis. The following topics are covered during the training:

• Triple quadrupole spectrometer calibration
• MRM pair formation and optimization
• Optimization of ion source parameters
• Optimization of chromatographic parameters
• Sample preparation using the example of blood drawn on Mitra® samplers (VAMS)
• Matrix effect, internal standards (ISTD), matrix curves
• Validation of the quantitative method
• Reporting of results

The goal of the training is to introduce users to the procedures and capabilities of the LC-MS/MS system and software to achieve sensitivity, selectivity, precision and reproducibility in quantitative analysis while covering the widest possible linear range of concentrations.

Training program

Day I

Theoretical part:

Triple quadrupole structure, ionization, MRM and sMRM scanning, chromatography, multimethods

Practical part:

• Q1/Q3 calibration (10 Da/s), MS/MS - resolution, m/z, intensity.
• MRM pair formation and optimization (manual, semi-automatic, automatic).
• Ion source optimization - T-concector and FIA
• Gradient vs isocrat, dosing, column selection, temperature selection, other important parameters
• Establishment and optimization of LC-MS/MS method.
• How to take care of MS cleanliness - ionization time programming, multi-way valve, cleaning of ion source and instrument front end

Sample preparation in pure solvent (standard curve, QC, solvent, solvent+ISTD, test samples), overnight analysis sequence.

Day II

Practical part - preparation of whole blood samples (Mitra®):

• Preparation of matrix standard curves
• Preparation of quality control (QC) samples in three concentrations
• Preparation of blank samples (without ISTD and with ISTD).
• Preparation of samples for recovery determination (“pre-spiked” and “post-spiked”)
• Preparation of test samples

Theoretical part:

• Samples in the matrix - discussion of the matrix effect and ways to level it.
• Optimization of sample preparation method and LC-MS/MS method

Practical part:

• Sample extraction from Mitra® samplers for LC-MS/MS analysis.
• Sequence of analysis of prepared samples overnight

Day III

Theoretical part:

• Validation of the quantitative LC-MS/MS method

Practical part:

• Software - data integration algorithms, signal smoothing, weighing.
• Matrix curves vs curves in solvent, influence of ISTD (Sciex OS).
• Method validation - validation criteria, determination of LOD, LLOQ, ULOQ, linearity, working range of the method, repeatability, recovery.
• Preparation of summary validation reports
• Reporting of results from test samples - report vs export of data to Excel.
• Summary and discussion.