Quantitative determination by liquid chromatography (LC) coupled with mass spectrometry
(MS) was achieved for the following 10 toxins found in association with diarrhetic
shellfish poisoning: okadaic acid (OA), dinophysistoxin-1 (DTX1), 7-O-palmitoylokadaic
acid (palOA), 7-O-palmitoyldinophysistoxin-1 (pa1DTX1), pectenotoxin-1 (PTX1), pectenotoxin-2
(PTX2), pectenotoxin-2 seco acid (PTX2SA), pectenotoxin-6 (PTX6), yessotoxin (YTX),
and 45-hydroxyyessotoxin (YTXOH). Toxins in 2 g of the adductor muscle or the digestive
glands of scallops, Patinopecten yessoensis, were extracted with 18 ml of methanol-water
(9:1, v/v), freed of polar contaminants by partition between chloroform and water,
and treated by solid-phase extraction on a silica cartridge column. Samples containing
YTXOH were purified separately on a buffered reversed-phase column. Chromatographic
separation was achieved by the following combinations of columns and mobile phases:
a Symmetry C18 column with acetonitrile-0.05% acetic acid (7:3, v/v) for OA, DTX1,
PTX6 and PTX2SA; a Develosil ODS column with the same mobile phase for PTX1 and PTX2;
a Capcellpak column with methanol-2.5% acetic acid (98:2, v/v) for palOA and palDTX1;
and an Inertsil ODS column with methanol-0.2 M ammonium acetate (8:2, v/v) for YTX
and YTXOH. Carboxylic acid toxins were selectively monitored on [M-H]- ions, sulfated
toxins on [M-Na]-ions, and neutral toxins on [M+NH4]+ ions. Average recoveries of
the toxins spiked to tissue homogenates ranged from 70 to 134%. Detection limits in
the muscle ranged from 5 to 40 ng/g and those in the digestive glands from 10 to 80
ng/g.