Fast and Sensitive Method for Determination of Domoic Acid in Mussel Tissue

Abstract

Domoic acid (DA), a neurotoxic amino acid produced by diatoms, is the main cause of amnesic shellfish poisoning (ASP). In this work, we propose a very simple and fast analytical method to determine DA in mussel tissue. The method consists of two consecutive extractions and requires no purification steps, due to a reduction of the extraction of the interfering species and the application of very sensitive and selective HILIC-MS/MS method. The procedural method was validated through the estimation of trueness, extract yield, precision, detection, and quantification limits of analytical method. The sample preparation was also evaluated through qualitative and quantitative evaluations of the matrix effect. These evaluations were conducted both on the DA-free matrix spiked with known DA concentration and on the reference certified material (RCM). We developed a very selective LC-MS/MS method with a very low value of method detection limit (9 ng g−1) without cleanup steps.

Toxicokinetics and Physiologically Based Pharmacokinetic Modeling of the Shellfish Toxin Domoic Acid in Nonhuman Primates

“…The lack of knowledge of the toxicokinetics of DA is largely due to limited bioanalytical methods to measure DA. LC-MS/MS methods exist for measuring DA in seawater and in contaminated seafood (Wang et al, 2012;Barbaro et al, 2016), but these methods lack the sensitivity needed to quantify DA in biological samples after low level exposure (Frame and Lefebvre, 2013). ELISA can measure DA at concentrations down to 0.4 ng/ml in biologic specimens (Frame and Lefebvre, 2013), but the ELISA also crossreacts with antibodies against DA, decreasing the usefulness of this assay following chronic exposures (Lefebvre et al, 2012).…”

The Preparation and Identification of a Monoclonal Antibody against Domoic Acid and Establishment of Detection by Indirect Competitive ELISA

“…Therefore, the results validated the working of ic-ELISA in complex matrix, showed insignificant levels of DA toxin found in the shellfish samples, and hence these samples were not contaminated and safe for the human consumption [ 8 ]. The ic-ELISA established in this study showed higher sensitivity demonstrating lower DA level in real shellfish samples (4 ng/g) compared with lower sensitivity exhibited previously by instrumental detection methods i.e., LC-MS/MS (9 ng/g) [ 37 ], LC-UV (6 ng/g) [ 38 ], respectively. Therefore, the results showed that the 1C3 mAb secreted by 1C3 hybridoma could be used to develop an ic-ELISA kit for the detection of DA.…”

Quantitative determination by screening ELISA and HPLC-MS/MS of microcystins LR, LY, LA, YR, RR, LF, LW, and nodularin in the water of Occhito lake and crops

“…Water samples processing and analysis Lake samples, previously analyzed by ELISA test, were also analyzed using HPLC-MS/MS instrumental method developed by Gambaro et al [23]. The pre-analytical procedure for lake water samples is quite similar than the protocol for a neurotoxin domoic acid in water samples developed by Barbaro et al [24]. An aliquot of lake water was acidified with 2 % of formic acid and was then spiked with 50 ng of the ENK internal standard in a 500-mL volumetric flask; the sample was then brought to volume with sample.…”