Streamlined Analysis of 400+ Pesticides in a Single Run Using the TSQ Quantum Access MAX Mass Spectrometer and TraceFinder Software

Importance of the Topic

Global food safety regulations increasingly demand high-throughput, sensitive multi-residue pesticide analysis. Regulatory frameworks in the European Union and Japan enforce maximum residue limits (MRLs) for hundreds of pesticides, driving the need for streamlined analytical workflows capable of detecting and confirming over 400 compounds in a single run.

Objectives and Study Overview

This study aimed to develop and validate a single-run LC-MS/MS method for simultaneous screening and quantitation of 437 pesticides within a 45-minute analysis. The approach integrates timed-selected reaction monitoring (T-SRM) and Quantitation-Enhanced Data-Dependent MS/MS (QED-MS/MS) for structural confirmation, with automated processing in TraceFinder software.

Methodology

Sample Preparation

• Standards prepared at 3 mg/L stock and diluted to 0.1–250 µg/L in acetonitrile/water (50:50).
• QuEChERS extraction of apple, orange, and asparagus matrices (15 g sample, 15 mL acetonitrile, dispersive cleanup), spiked at 2 µg/kg for recovery tests.

Chromatographic Conditions

• Column: Hypersil GOLD aQ, 100 × 2.1 mm, 1.9 µm.
• Mobile phases: A = water + 0.1% formic acid + 4 mM ammonium formate; B = methanol + 0.1% formic acid + 4 mM ammonium formate.
• Flow rate: 300 µL/min; gradient from 2% to 100% B over 35 min; column at 40 °C; injection 10 µL.

Used Instrumentation

• Thermo Scientific TSQ Quantum Access MAX triple quadrupole MS with heated electrospray ionization (HESI).
• Thermo Scientific Accela 1250 U-HPLC system with HTC-PAL autosampler.
• TraceFinder software for method development, data acquisition, review, and reporting.

Main Results and Discussion

T-SRM scheduling enabled monitoring of 933 transitions with 60 s windows, improving duty cycle and sensitivity. Limits of detection ranged from 0.1 to 50 µg/L, with correlation coefficients above 0.98 (most > 0.99). All 437 pesticides were chromatographically separated and quantified in 45 min. QED-MS/MS provided fragment-rich spectra for positive structural confirmation. Spiked apple, orange, and asparagus matrices at 2 µg/kg yielded clear chromatograms and passed automated ion ratio criteria.

Benefits and Practical Applications of the Method

This workflow supports regulatory compliance and routine QA/QC by offering

• High throughput: over 400 analytes per run.
• Sensitivity and selectivity: low detection limits and structural confirmation.
• Automated data processing: flagging, re-integration, and report generation.

Future Trends and Potential Applications

Advances may include expanding compound libraries, integrating high-resolution MS for non-target screening, further automating sample preparation, and real-time field monitoring. The approach can be adapted to emerging contaminants and diverse food matrices.

Conclusion

The combination of timed-SRM, QED-MS/MS, and TraceFinder software delivers a robust, efficient, and sensitive solution for large-scale multi-residue pesticide analysis, meeting stringent MRL requirements and facilitating rapid decision-making in food safety laboratories.

References

1. Ministry of Health, Labour and Welfare Japan Positive List System introduction 2006
2. Regulation (EC) No 396/2005 of the European Parliament and of the Council on pesticide residues in food
3. EU Directive 91/414/EEC pesticide safety review program
4. Lehotay SJ Quick Easy Cheap Effective Rugged and Safe (QuEChERS) Approach for Determining Pesticide Residues Methods in Biotechnology 2006 19 239–261