Liquid Chromatography with Mass Spectrometry—Detection of Lipophilic Shellfish Toxins

Abstract

A rapid multiple toxin method based on liquid chromatography with mass spectrometry (LC/MS) was developed for the detection of okadaic acid (OA), dinophysistoxin-1 (DTX-1), DTX-2, yessotoxin (YTX), homoYTX, 45-hydroxy-YTX, 45-hydroxyhomo-YTX, pectenotoxin-1 (PTX-1), PTX-2, azaspiracid-1 (AZA-1), AZA-2, and AZA-3. Toxins were extracted from shellfish using methanol–water (80%, v/v) and were analyzed using a C8 reversed-phase column with a 5 mM ammonium acetate–acetonitrile mobile phase under gradient conditions. The method was validated for the quantitative detection of OA, YTX, PTX-2, and AZA-1 in 4 species (mussels, Mytilus edulis; cockles, Cerastoderma edule; oysters, Crassostrea gigas; king scallop, Pecten maximus) of shellfish obtained from United Kingdom (UK) waters. Matrix interferences in the determination of the toxins in these species were investigated. The validated linear range of the method was 13–250 μg/kg for OA, PTX-2, and AZA-1 and 100–400 μg/kg for YTX. Recovery and precision ranged between 72–120 and 1–22%, respectively, over a fortification range of 40–160 μg/kg for OA, PTX-2, and AZA-1 and 100–400 μg/kg for YTX. The limit of detection, reproducibility, and repeatability of analysis showed acceptable performance characteristics. A further LC/MS method using an alkaline hydrolysis step was assessed for the detection of OA, DTX-1, and DTX-2 on their esterified forms. In combination with the LC/MS multiple toxin method, this allows detection of all toxin groups described in Commission Decision 2002/225/EC.

Yessotoxins, a Group of Marine Polyether Toxins: an Overview

“…It is possible to analyze all the YTXs, even those derivatized with DMEQ-TAD [ 108 ], and also different algal toxins. In fact, several multi-toxin detection methods have been developed [ 109 ]. However, as is the case for other chemical methods, calibration standards are required for method development and toxin quantification.…”

Determination of lipophilic marine toxins in mussels. Quantification and confirmation criteria using high resolution mass spectrometry

“…However, with this technique it is mandatory to detect compounds that are pre-selected and it is sensible to matrix effects. Several methods of multitoxin analysis are described in the literature [13][14][15][16]. Some of them are intra-or inter-laboratory validated [15,17,18], but the analysis of lipophilic toxins by means of mass spectrometry still generates some controversies [19][20][21][22].…”

Marine biotoxins in shellfish – Pectenotoxin group

“…A collaborative study by McNabb et al (2005) focused on the analysis of extracts and found good apparent recovery. Stobo et al (2005), found little matrix effect in the determination of PTX2 by LC-MS, while Fux et al (2008) found between 0 and 92 % signal enhancement, depending on chromatographic conditions. A study by Gerssen et al (2009a) also confirmed that reproducibility is poor for PTX2 compared to other compounds.…”