New method for the analysis of lipophilic marine biotoxins in fresh and canned bivalves by liquid chromatography coupled to high resolution mass spectrometry: A quick, easy, cheap, efficient, rugged, safe approach

• A QuEChERS methodology is applied in the lipophilic marine biotoxins (MBTXs) analysis.
• UHPLC with HRMS/MS furnishes excellent results in the quantitative analysis of MBTXs.
• Calibration with surrogate matrix matched standards provides reliable results.
• Eprinomectin is proposed as internal standard for lipophilic marine biotoxins.
• The method is suitable for the analysis of both fresh and canned bivalve molluscs.

A new method for the analysis of lipophilic marine biotoxins (okadaic acid, dinophysistoxins, azaspiracids, pectenotoxins, yessotoxins, spirolids) in fresh and canned bivalves has been developed. A QuEChERS methodology is applied; i.e. the analytes are extracted with acetonitrile and clean-up of the extracts is performed by dispersive solid phase extraction with C18. The extracts are analyzed by ultra-high performance liquid chromatography coupled to a hybrid quadrupole-Orbitrap mass spectrometer, operating in tandem mass spectrometry mode, with resolution set at 70,000 (m/z 200, FWHM). Separation of the analytes, which takes about 10 min, is carried out in gradient elution mode with a BEH C18 column and mobile phases based on 6.7 mM ammonia aqueous solution and acetonitrile mixtures. For each analyte the molecular ion and 1 or 2 product ions are acquired, with a mass accuracy better than 5 ppm. The quantification is performed using surrogate matrix matched standards, with eprinomectin as internal standard. The high-throughput method, which has been successfully validated, fulfills the requirements of European Union legislation, and has been implemented as a routine method in a public health laboratory.