Hydrophilic interaction liquid chromatography/mass spectrometry for determination of domoic acid in Adriatic shellfish

Abstract

This paper describes a new method for sensitive, specific and direct determination of domoic acid (DA), the causative toxin of amnesic shellfish poisoning (ASP) syndrome, in shellfish. It is based on combination of hydrophilic interaction liquid chromatography with mass spectrometry (HILIC/MS). The high percentage of organic modifier in the mobile phase and the omission of ion-pairing reagents, both favoured in HILIC, result in enhanced detection limits with MS detection. The new method was set up either on an ionspray ion trap MS instrument operating in MS and MS/MS scanning acquisition modes, or on a turboionspray triple-quadrupole MS system operating in selected ion monitoring (SIM) and multiple reaction monitoring (MRM) acquisition modes. Positive and negative ion experiments were performed. MRM experiments are recommended for screening contaminated shellfish tissue and for quantitative analyses due to highest sensitivity and selectivity. The minimum detection levels for the toxin in tissue were found to be 63 and 190 ng/g in positive and negative MRM experiments, respectively, which are well below the regulatory limit for DA in tissue (20 microg/g). Application to shellfish samples collected in the Adriatic Sea (Italy) in the period 2000-2004 demonstrated for the first time in Italy the presence of DA as a new toxin that has entered the Adriatic Mytilus galloprovincialis toxin profile.

Hydrophilic interaction chromatography

“…The method is robust and analyses can be done without sample cleanup or preconcentration. It was later extended to domoic acid [165] and a wider range of PSP toxins [166], and recently applied along with other methods for studies of the toxin profile of Mytilus galloprovincialis [167].…”

“…Certain solutes seem to be more affected than others. [54,233,266]; pyridylaminated sialo sugar chains [219]; quaternary ammonium compounds [133]; small polar compounds in food analysis by HILIC/ESI-MS [155]; hydroxybenzoate saxitoxin analogs in dinoflagellates [160]; chitine fragments [198]; cyanobacterial toxins by HILIC/MS [164]; 1-deoxynojirimycin in mulberry leaves [150]; polar compounds contributing to umami taste and mouth-drying oral sensation of morel mushrooms [156]; elucidation of yeast sulfur metabolism [253]; PSP toxins [166,165,167]; monosaccharides [227,267]; identification of a bitter inhibitor [157] Peptides, nucleic acids, and other polar compounds [13]; peptides [67]; tyrosine protein-kinase specificity [234]; desalting of electroeluted proteins [241]; glycopeptides [268]; site-specific glycosylation microheterogeneity in recombinant human interferon-c [269]; urea, allantoin and lysine pyroglutamate in cosmetic samples [134]; glucosinolates in vegetables [201]; folates in human plasma by HILIC/ MS/MS [137]; citrulline in dried blood spots [138]; SPE enrichment of avoparcin from kidney samples [254]: Ionic compounds by pressurized flow CEC [270]; sucrose in a plasma-derived process solution [271]; glycoalkaloids in transgenic potatoes [104]; glycopeptide microheterogeneity [272] Peptides [67,68,276]; amphipathic a-helical peptides …”

The advantages of ESI‐MS detection in conjunction with HILIC mode separations: Fundamentals and applications

“…Multi-toxin determination was possible even at trace levels [146,149], allowing a rapid, simultaneous screening of a wide range of toxins [145]. A specific compound related to toxins, domoic acid, was also determined at low detection levels in Adriatic shellfish samples [149]. Within these applications, amide columns with an ammonium salt additive in the mobile phase seemed to be commonly exerted in HILIC separation with ESI-MS/MS detection.…”

“…To obtain the more complete profiles of toxins from a specific species, several research groups assimilated data from HILIC-ESI-MS and from RPLC on a C 8 column coupled with ESI-MS [146,147] or on a C 18 column with post-column derivatization-fluorescence detection [148]. Multi-toxin determination was possible even at trace levels [146,149], allowing a rapid, simultaneous screening of a wide range of toxins [145]. A specific compound related to toxins, domoic acid, was also determined at low detection levels in Adriatic shellfish samples [149].…”

Palytoxins: A still haunting Hawaiian curse

“…Since our group had long been involved in the isolation, analysis, and structural determination of marine biotoxins, we were asked by ARPAL (Ligurian Regional Agency for the Environmental Protection) to investigate plankton samples collected off Genoa during the toxic outbreak. Firstly, the presence of okadaic acid, spirolides, yessotoxins, saxitoxins, and domoic acid in the Genoa extract was investigated by LC-MS (Quilliam et al 2001;Dell'Aversano et al 2005;Ciminiello et al 2005). But none of the above toxins-selected because quite common across the Mediterranean Sea (Ciminiello and Fattorusso 2004)-was indeed detected.…”