Selective liquid chromatographic determination of trace domoic acid in seawater and phytoplankton: improvement using the o‐phthaldialdehyde/9‐fluorenylmethylchloroformate derivatization

Abstract

Domoic acid (DA), the amnesic shellfish poisoning toxin (ASP), is an excitatory amino acid that can accumulate, under certain environmental conditions, in shellfish, finfish, birds, and mammals, by direct filtration or by feeding on contaminated organisms (Mos 2001). This potent neurotoxin with three carboxylic groups, responsible for its high polarity and hydrophilicity, is a cyclic amino acid and a secondary amine with a molecular weight of 311 Daltons. DA belongs to a neurotransmitter class of compounds with a structure very similar to an important neurotransmitter excitatory, acid glutamate, and indeed mimics glutamate in its interaction with some of its receptor subtypes. DA can damage the neurons by activating R-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and kainate receptors, causing an influx of calcium, and then be the cause of several symptoms such as nausea, disorientation, temporary amnesia and, in more serious cases, persistent memory loss and/or coma, and ultimately even death (Costa et al. 2010). This toxin was originally isolated from a red microalga Chondria armata by Japanese researchers (Takemoto and Daigo 1958) and is now reported to be produced in several species of marine diatoms from the genus Pseudo-nitzschia. DA was identified as the causative agent of the well-known tragic intoxication event observed in 1987 (Prince Edward Island from Cardigan Bay in Eastern Canada). More than one hundred people became ill and three victims died, by septic shock or pneumonia, 11-24 days after intoxication. Whereas most victims recovered within 10 days, after a period of confusion and nausea, the others continued to show signs of selective short-term memory loss (Bates et al. 1989). Additional consequences of the DA presence included the temporary closure of shellfish aquaculture industries and a strong impact on tourism activities (Pistocchi et al. 2012;Trainer et al. 2012).The detection limit of the method generally considered as the reference for particulate DA quantification in marine organisms (high-performance liquid chromatography with ultra-violet detection at λ = 242 nm, HPLC-UV) ranges between 13 and 250 nM depending on the detector (Quilliam 2003). This sensitivity threshold is often insufficient to follow the dynamics of both dissolved and particulate domoic acid AbstractDomoic acid (DA), a toxin produced worldwide by some species of the genus Pseudo-nitzschia, is responsible for contamination of marine molluscs, mammals, birds, and for human intoxication, and when detected in high levels results in closures of shellfish farms, thus causing severe economic losses to aquaculture. Studies on algal production of DA in cultures and field samples require sensitive methods capable of measuring trace concentrations of domoic acid. Measuring domoic acid concentrations at trace levels is still a significant challenge. A sensitive and reliable double derivatization using o-phthaldialdehyde-mercaptoethanol/9-fluorenylmethylchloroformate (OPA-MeSH/FMOC-Cl) followed by high-performance liq...

Diversity and regional distribution of harmful algal events along the Atlantic margin of Europe

“…At least 153 people were affected with symptoms including both diarrheic and neurological, including confusion and memory loss and there were three mortalities (Quilliam et al, 1988Quilliam and Wright, 1989;Perl et al, 1990). This event led to the determination of domoic acid (DA) as the causative toxin and the subsequent rapid development of monitoring methods using liquid chromatography (LC) with UV or diode-array detection (DAD), for shellfish tissues, and shortly thereafter a more sensitive fluorescence-based method (LC-FD) for detection in plankton and seawater matrices (Pocklington et al, 1990, Devez andDelmas 2013). These chemical analytical methods were implemented into an ongoing regulatory management programme from the early 1990s for prevention of subsequent outbreaks affecting human health of shellfish consumers (Quilliam et al, 1991).…”

A Rapid LC-HRMS Method for the Determination of Domoic Acid in Urine Using a Self-Assembly Pipette Tip Solid-Phase Extraction

“…Therefore, DA is considered a significant risk to public health and to the economic viability of the aquaculture and shellfish harvesting industries. To ensure human health protection, monitoring programs are carried out in many countries and several analytical methods have been developed for the monitoring of DA in shellfish, phytoplankton, and seawater [ 7 , 8 , 9 , 10 , 11 , 12 , 13 ]. The European Community’s regulations have required the concentration of DA in shellfish should not exceed the permissible limit of 20 mg/kg [ 14 ].…”

A surface plasmon resonance system for the underwater detection of domoic acid

“…It would then be possible to work with lower antibody concentrations. Even though our system demonstrates very good performances, the relative standard deviation of the measurements is about 15%, which is two to three times greater than with chemical methods (Wang et al 2007;Devez and Delmas 2013).…”