Bioanalysis of Ametryn by UHPLC-MS³ Coupled With Multiple Fragmentation to Decrease Interference and Enhance...

Area of Science:

• Environmental Chemistry
• Analytical Chemistry
• Toxicology

Background:

• Ametryn poses risks to aquatic ecosystems.
• Conventional methods for ametryn detection face challenges with matrix interference and sensitivity.
• There is a need for improved analytical techniques for ametryn quantification in biological samples.

Purpose of the Study:

• To develop and validate a highly selective and sensitive UHPLC-MS^3 method for ametryn quantification.
• To apply the developed method for analyzing ametryn levels in various zebrafish tissues.
• To establish a reliable platform for ametryn residue monitoring and toxicological research.

Main Methods:

• A novel ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS^3) method utilizing electrospray ionization was established.
• Quantification was performed using the MS^3 transition m/z 228.0 → 186.1 → 158.0.
• Sample preparation and analysis were conducted on five zebrafish tissues (liver, muscle, brain, intestine, gill) with rapid chromatographic separation (2.8 min).

Main Results:

• The UHPLC-MS^3 method significantly reduced matrix interference compared to MRM.
• Signal intensity increased from 6.4 × 10^4 to 1.6 × 10^7 cps, and signal-to-noise ratio improved from 6.3 to 45.
• The assay demonstrated excellent linearity (10-1000 pg/mL) and met FDA bioanalytical requirements for accuracy, precision, recovery, matrix effects, and stability.

Conclusions:

• This study successfully implemented UHPLC-MS^3 for ametryn quantification in multiple zebrafish tissues.
• The developed method provides a reliable and high-throughput platform for ametryn residue monitoring.
• This technique supports mechanistic toxicology research by enabling accurate ametryn level determination in biological samples.