MassHunter MRM/dMRM/tMRM Database - Familiarization Guide

Význam tématu

Targeted multiple-reaction monitoring (MRM) approaches on triple quadrupole LC/MS systems enable highly selective and sensitive quantitation of trace analytes in complex matrices. The incremental workflows MRM → dynamic MRM (dMRM) → triggered MRM (tMRM) allow analysts to maximize dwell time, minimize cycle time, and confirm compound identity via secondary transitions and retention time windows. This methodology is critical in environmental, food safety, and pharmaceutical QA/QC applications requiring robust pesticide screening and confirmation.

Cíle a přehled studie / článku

This guide demonstrates how to use the Agilent MassHunter MRM/dMRM/tMRM Database to develop and verify MRM, dMRM, and tMRM methods. Using the Checkout Mix pesticide standard, it covers:

• Importing compound and transition parameters from a curated database into an MRM acquisition method
• Acquiring and inspecting MRM data in the Qualitative Analysis software to assign retention times
• Converting the MRM method to a dMRM method using quantified retention time windows and cycle time optimization
• Extending the method to tMRM by adding trigger thresholds, entrance delays, and secondary confirmation transitions
• Building a reference library for spectral matching in Quantitative Analysis

Použitá metodika a instrumentace

• LC/MS system: Agilent 1200/1260 UHPLC with ZORBAX Eclipse Plus C18 column (2.1×100 mm, 1.8 µm) and Agilent 6400 Series Triple Quadrupole MS
• Software: MassHunter Data Acquisition B.06.00+, Qualitative Analysis B.07.00+, Quantitative Analysis B.07.01+
• Mobile phases: A – 5 mM acetic acid in water; B – acetonitrile, gradient from 5 % to 95 % B over 12 min
• Method development: database import of compound names, precursor/product transitions, fragmentor voltages, collision energies; dynamic scheduling based on measured peak widths

Hlavní výsledky a diskuse

• MRM import from the database produced a consistent initial method, with primary and qualifier transitions for 14 pesticides
• dMRM update optimized retention time windows (±1 min baseline) and cycle time (500 ms) to ensure ≥10 data points across 5 s peaks, with dwell times >5 ms
• tMRM scheduling with trigger thresholds, entrance/trigger delays and window of 0.5 min focused secondary transition acquisition near peak apex
• Qualitative Analysis confirmed acquisition of all primaries and secondaries; a reference library built in Quantitative Analysis yielded 100 % match scores for standards

Přínosy a praktické využití metody

By applying database-driven MRM method creation and dynamic scheduling, laboratories can achieve rapid method development and high throughput screening with improved specificity. Triggered MRM adds confirmation power without sacrificing sensitivity, making it ideal for regulatory compliance and trace-level quantitation in environmental and food matrices.

Budoucí trendy a možnosti využití

• Integration of real-time feedback algorithms and AI-driven retention time prediction for automated scheduling
• Expansion to high-multiplex panels (>200 analytes) using advanced agile scheduling and variable dwell time
• Application in metabolomics, biomarker validation, and clinical assays requiring high selectivity and throughput
• Improved data analytics and spectral libraries for reproducible confirmatory workflows

Závěr

The Agilent MassHunter MRM/dMRM/tMRM Database streamlines targeted method creation by importing validated compound and transition parameters. The described workflow—MRM method import, dMRM dynamic scheduling, and tMRM triggered confirmation—provides a systematic approach for high-confidence targeted quantitation and confirmation of pesticide analytes in complex samples. This flexible strategy supports rapid method adaptation and robust performance across various triple quadrupole platforms.

Reference

• Agilent Technologies. MassHunter MRM/dMRM/tMRM Database Familiarization Guide, Rev. A, March 2016
• Agilent Technologies. MassHunter Data Acquisition B.06.00 User Manual
• Agilent Technologies. MassHunter Quantitative and Qualitative Analysis B.07 User Guides