Liquid chromatography with electrospray ion-trap mass spectrometry for the determination of yessotoxins in shellfish

Yessotoxins are a group of large polyether toxins, produced by marine dinoflagellates, which cause widespread contamination of filter-feeding shellfish. A new, sensitive liquid chromatography–mass spectrometry (LC–MS) method has been developed for the determination of yessotoxin (YTX) and 45-hydroxy-yessotoxin (45-OHYTX), a major metabolite in shellfish. The LC system was coupled, via an electrospray ionisation (ESI) source, to an ion-trap MS in negative mode. The molecular related ion species at m/z 1141 [M-2Na+H]− was used as the parent ion for multiple MS experiments. MS–MS and MS3 gave major fragment ions at m/z 1061 [1141-SO3H]− and m/z 945 [1061-C9H12O]−. Predominant ions, that are due to the fragmentation of the backbone structure of YTXs, were observed at the MS4 stage. Reversed-phase LC using a C16 amide column was preferable to C18 phases for the separation of YTX and 45-OHYTX. Optimum calibration and reproducibility data were obtained for YTX using LC–MS–MS; r2=0.9960, RSD≤6.3% at 0.25 μg YTX/g (n=5). The detection limit (S/N=3) was 30 pg YTX on-column which corresponded to 3 ng/g shellfish tissue.