Analysis of anatoxin-a in biological samples using liquid chromatography with fluorescence detection after solid...

Area of Science:

• Environmental Chemistry
• Analytical Chemistry
• Toxicology

Background:

• Anatoxin-a is a potent neurotoxin produced by cyanobacteria in freshwater environments.
• This toxin poses a significant health risk, particularly to animals, necessitating reliable detection methods.

Purpose of the Study:

• To develop and validate a sensitive High Performance Liquid Chromatography with Fluorescence detection (HPLC-FLD) method for anatoxin-a determination.
• To assess the method's efficacy across diverse biological matrices, including cyanobacterial cultures, water, and fish/mussel tissues.

Main Methods:

• Intracellular anatoxin-a extraction using sonication.
• Sample concentration and purification via solid-phase extraction (SPE) with weak cation exchange.
• Derivatization of anatoxin-a with 4-Fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) for fluorescence detection.

Main Results:

• The developed HPLC-FLD method demonstrated adequate recovery rates (71-87%) across tested matrices.
• Low limits of detection were achieved: ng/L for water samples and ng/g for cyanobacterial samples.
• Comparison with solid-phase microextraction (SPME) highlighted differences in impurity removal, simplicity, rapidity, and concentration enhancement.

Conclusions:

• The validated HPLC-method provides a sensitive and reliable approach for quantifying anatoxin-a in environmental and biological samples.
• The study discusses the comparative advantages of SPE and SPME for anatoxin-a analysis, aiding method selection.
• This research contributes to safeguarding public and animal health by enabling effective monitoring of cyanobacterial toxins.