Automated on-line solid-phase extraction coupled to liquid chromatography-tandem mass spectrometry for determination of lipophilic marine toxins in shellfish.

Automated on-line solid-phase extraction (SPE) coupled to liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been developed for fast determination of lipophilic marine toxins in shellfish samples. The direct coupling of an on-line SPE column to LC-MS/MS was accomplished using column switching techniques. Suitable chromatographic separation was performed on a reversed-phase C18 column under alkaline conditions (pH 11). The selected reversed-phase C18 SPE column allowed rapid and efficient on-line desalting of hydrolysed shellfish samples, avoiding signal suppression during mass spectrometry detection. Furthermore, the on-line SPE procedure allowed reducing matrix effects observed in raw and hydrolysed shellfish extracts. The proposed method was evaluated in terms of linearity, precision, accuracy and limits of detection (LODs). Quantitative recovery (97-102%) and satisfactory inter-day precision (RSD<8%) were achieved for all target compounds. LODs in the sub-μgkg level (0.37-0.68μgkg) were obtained for all toxins except for okadaic acid, which showed a value of 2.75μgkg. Several mussel samples from North-western Spain were finally analysed in order to demonstrate the applicability of the method. Okadaic acid was the predominant toxin in all samples, although other lipophilic toxins were also detected.