Coupling High-Throughput Magnetic-Bead-Based Immunoaffinity with LC-MS/MS Analysis for Quantifying Cylindrospermopsin...

Area of Science:

• Environmental Chemistry
• Analytical Chemistry
• Toxicology

Background:

• Cylindrospermopsin (CYN) bioaccumulation in aquatic food chains poses risks to human and animal health.
• Complex biological matrices challenge accurate LC-MS/MS analysis of CYN.
• Efficient CYN enrichment methods are crucial for toxicological studies.

Purpose of the Study:

• To develop a novel immunoaffinity extraction method for CYN enrichment.
• To overcome matrix effects in LC-MS/MS analysis of CYN.
• To assess CYN and deoxy-CYN levels in fish tissues.

Main Methods:

• Utilized magnetic beads functionalized with anti-CYN monoclonal antibodies for specific CYN capture.
• Developed an automated extraction process compatible with LC-MS/MS.
• Validated the method for sensitivity, linearity, precision, and recovery.

Main Results:

• The automated immunoaffinity method processed 32 samples in 54 minutes without centrifugation or filtration.
• Achieved high sensitivity (LOQ 0.025 μg L^-1), linearity (R² > 0.99), precision (RSD < 6%), and recovery (85-105%).
• Detected deoxy-CYN more frequently and at higher concentrations than CYN in fish tissues, with significant tissue-specific accumulation (muscle: 2.38 μg kg^-1, liver: 71.99 μg kg^-1).

Conclusions:

• This study presents the first report of immunoaffinity extraction for CYN enrichment.
• The developed method is reliable, automated, and effective in mitigating matrix effects for LC-MS/MS analysis.
• The findings highlight the prevalence of deoxy-CYN and tissue-specific accumulation patterns of CYN analogues in fish.