Simultaneous Suspect Screening and Quantitation of Pesticides in a Complex Matrix Using an Agilent 6546 LC/Q-TOF

Significance of the topic

Pesticide contamination in complex food matrices such as black pepper poses a significant risk to consumer health and global trade. Black pepper’s rich composition of essential oils, alkaloids, and pigments complicates analytical detection of trace residues. High resolution mass spectrometry coupled with efficient sample cleanup is critical to meet stringent regulatory maximum residue limits and to ensure reliable screening for a wide range of pesticide chemistries.

Objectives and Overview

This application note describes a comprehensive workflow for simultaneous suspect screening and targeted quantitation of over 300 pesticides in black pepper. The aim was to develop a single analytical method that provides high throughput, robust cleanup, and accurate quantification to satisfy regulatory and quality control requirements.

Methodology and Instrumentation

Samples were extracted using an Agilent Bond Elut QuEChERS EN kit, followed by pass-through cleanup with Agilent Captiva EMR–GPD cartridges. Chromatographic separation employed an Agilent 1290 Infinity II UHPLC, and detection used an Agilent 6546 LC/Q-TOF MS operated in All Ions MS/MS data-independent acquisition mode. Agilent MassHunter Quantitative Analysis software with a personal compound database library enabled integrated workflows for both targeted quantitation and suspect screening.

Main results and discussion

The method achieved screening detection limits of ≤5 µg/kg for most pesticides. Limits of quantitation met regulatory thresholds at ≤10 µg/kg, with calibration linearity above R2 = 0.99 across 0.5–10 µg/L in matrix-matched standards. Mean recoveries for the majority of analytes ranged from 70 to 120 %, with intraday and interday precision below 20 % RSD. Mass accuracy was consistently within ±5 ppm throughout the chromatographic run, ensuring reliable identification of both target and suspect compounds.

Benefits and practical applications of the method

The unified workflow reduces analysis time by combining targeted and non-target screening in a single instrument run. Efficient cleanup improves instrument performance and minimizes matrix effects. This approach supports routine QA/QC and regulatory testing laboratories in maintaining high throughput without sacrificing data quality.

Future trends and potential applications

The methodology can be extended to other challenging food and botanical matrices. Emerging cleanup materials and advanced data processing algorithms, including machine learning-enhanced screening, will further expand non-targeted contaminant discovery. Integration with automated sample preparation platforms promises additional gains in throughput and reproducibility.

Conclusion

The described workflow delivers comprehensive suspect screening and accurate quantitation of hundreds of pesticides in black pepper using a single LC/Q-TOF analysis. The combination of tailored sample preparation, high resolution mass spectrometry, and unified data processing meets the stringent performance criteria required for modern food safety testing.

References

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• European Commission EU Pesticide Residues Database 2022
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• Pham TQ et al Simultaneous Screening and Quantification of Pesticide Residues in Red Cabbage Agilent Application Note 5994-3123EN 2021
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• Yannell KE, Chen K Enhanced Food Safety Testing A Pesticide Screening Methodology Using the Agilent 6546 LC/Q-TOF and MassHunter Screener Agilent Application Note 5994-0738EN 2019