Development of a validated UPLC-MS³ method for trace-level azoxystrobin determination in zebrafish liver tissues.

Area of Science:

• Environmental Science
• Analytical Chemistry
• Toxicology

Background:

• Azoxystrobin, a strobilurin fungicide, poses environmental risks due to its persistence and aquatic toxicity.
• Accurate quantification of azoxystrobin in biological tissues is crucial for environmental risk assessment.
• Existing methods struggle with sensitivity and specificity for trace-level determination in small tissue samples.

Purpose of the Study:

• To develop and validate a highly sensitive and specific analytical method for trace azoxystrobin determination in zebrafish liver.
• To establish a robust platform for studying fungicide biodistribution in aquatic organisms.
• To enhance environmental toxicology research through precise fungicide quantification.

Main Methods:

• Ultra-performance liquid chromatography coupled with triple-stage mass spectrometry (UPLC-MS3).
• Protein precipitation extraction followed by C18 column chromatographic separation.
• Optimized MS3 transition (m/z 404.0 → 371.9 → 344.2) for detection.

Main Results:

• The UPLC-MS3 method demonstrated excellent linearity (r > 0.9984) and accuracy (-3.33% to 3.67%).
• High precision (CV 5.57-10.19%), consistent recoveries (94.93-106.64%), and minimal matrix effects (99.45-104.79%) were achieved.
• MS3 significantly improved specificity over conventional MRM by reducing interference, enabling successful application in zebrafish tissue distribution studies.

Conclusions:

• The validated UPLC-MS3 method provides a sensitive and specific approach for trace azoxystrobin quantification in zebrafish liver.
• This method is reliable for environmental toxicology research and investigating fungicide biodistribution in aquatic ecosystems.
• The developed technique offers a robust platform for future studies on fungicide impacts in aquatic organisms.