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

2016 ◽  
Vol 2016 ◽  
pp. 1-6 ◽  
Author(s):  
Elena Barbaro ◽  
Roberta Zangrando ◽  
Carlo Barbante ◽  
Andrea Gambaro
Keyword(s):  
Analytical Method ◽  
Domoic Acid ◽  
Shellfish Poisoning ◽  
Method Detection Limit ◽  
Mussel Tissue ◽  
Qualitative And Quantitative ◽  
Amnesic Shellfish Poisoning ◽  
The Matrix ◽  
Procedural Method

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.

scholarly journals Determination of Domoic Acid in Japanese Mussels by Enzyme Immunoassay

2000 ◽  
Vol 83 (6) ◽  
pp. 1384-1386 ◽  
Author(s):  
Kentaro Kawatsu ◽  
Yonekazu Hamano ◽  
Tamao Noguchi
Keyword(s):  
Monoclonal Antibody ◽  
Enzyme Immunoassay ◽  
Domoic Acid ◽  
Immunoaffinity Chromatography ◽  
Shellfish Poisoning ◽  
Mussel Tissue ◽  
Blue Mussels ◽  
Amnesic Shellfish Poisoning ◽  
Low Concentrations

Abstract Ten samples of commercial blue mussels (Mytilus edulis) from Japan were analyzed for domoic acid by an indirect competitive enzyme immunoassay (idc–EIA) based on an anti-domoic acid monoclonal antibody. Domoic acid was found in all samples at low concentrations (0.11–1.81 ng/g mussel tissue). The presence of domoic acid was confirmed by liquid chromatography coupled with immunoaffinity chromatography using an anti-domoic acid monoclonal antibody as ligand. To our knowledge, this is the first reported detection of domoic acid, a causative agent of amnesic shellfish poisoning, in Japanese mussels.

scholarly journals Sensitive determination of domoic acid in mussel tissue using dansyl chloride derivatization and liquid chromatography-mass spectrometry

2015 ◽  
Vol 7 (3) ◽  
pp. 1000-1007 ◽  
Author(s):  
Daniel G. Beach ◽  
Hechun Liu ◽  
Michael A. Quilliam
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Domoic Acid ◽  
New Method ◽  
Liquid Chromatography Mass Spectrometry ◽  
Shellfish Poisoning ◽  
Mussel Tissue ◽  
Amnesic Shellfish Poisoning ◽  
Sensitive Determination

This paper describes a new method for sensitive determination of domoic acid (DA), the causative toxin of amnesic shellfish poisoning (ASP), in shellfish.

scholarly journals Determination and Confirmation of the Amnesic Shellfish Poisoning Toxin, Domoic Acid, in Shellfish from Scotland by Liquid Chromatography and Mass Spectrometry

2001 ◽  
Vol 84 (5) ◽  
pp. 1657-1667 ◽  
Author(s):  
Philipp Hess ◽  
Susan Gallacher ◽  
Lesley A Bates ◽  
Nigel Brown ◽  
Michael A Quilliam
Keyword(s):  
Liquid Chromatography ◽  
Domoic Acid ◽  
Pecten Maximus ◽  
Shellfish Poisoning ◽  
Mussel Tissue ◽  
Crude Extracts ◽  
Amnesic Shellfish Poisoning ◽  
Routine Monitoring ◽  
Strong Anion Exchange ◽  
Array Detection

Abstract During 1998 and early 1999, shellfish samples from sites in Scotland were found to contain the amnesic shellfish poisoning toxin, domoic acid (DA). Two different techniques, liquid chromatography (LC) with UV diode-array detection and LC with mass spectrometric (MS) detection, were used to detect and confirm DA in shellfish extracts. The LC/UV method was validated for routine monitoring by recovery experiments on spiked mussel and scallop tissues with a certified mussel tissue used as reference material. Crude extracts of selected samples as well as extracts cleaned with strong anion exchange (SAX) were analyzed by both LC/UV and LC/MS. Good correlation (linear regression r2 = 0.996, slope = 0.93) between the 2 methods was found for cleaned extracts. Analyses of crude extracts by LC/UV produced false-positive results in 2 crab samples, whereas LC/MS analyses gave accurate results. It was concluded that LC/UV is a valid approach for routine monitoring of DA in shellfish when cleanup is performed with a SAX cartridge to prevent false positives. A variety of shellfish species were surveyed for DA content, including Pecten maximus (king scallops), Chlamys opercularis (queen scallop), Mytilus edulis (blue mussels), Cancer pugaris (crab), and Ensis ensis (razor fish). The highest concentration of DA was 105 μg/g in Pecten maximus.

scholarly journals Long Term Memory Outcome of Repetitive, Low-Level Dietary Exposure to Domoic Acid in Native Americans

2021 ◽  
Vol 18 (8) ◽  
pp. 3955
Author(s):  
Lynn M. Grattan ◽  
Laura Kaddis ◽  
J. Kate Tracy ◽  
John Glenn Morris
Keyword(s):  
Native Americans ◽  
Domoic Acid ◽  
Dietary Exposure ◽  
Long Term Memory ◽  
Shellfish Poisoning ◽  
Amnesic Shellfish Poisoning ◽  
The Pacific ◽  
Small Decline ◽  
Speed And Accuracy

Domoic acid (DA) is a marine-based neurotoxin that, if ingested via tainted shellfish, is associated with Amnesic Shellfish Poisoning (ASP). These acute effects of elevated DA exposure in humans have been well described. In contrast, the long-term impacts of lower level, repetitive, presumably safe doses of DA (less than 20 ppm) are minimally known. Since Native Americans (NA) residing in coastal communities of the Pacific NW United States are particularly vulnerable to DA exposure, this study focuses on the long-term, 8-year memory outcome associated with their repeated dietary consumption of the neurotoxin. Measures of razor clam consumption, memory, clerical speed and accuracy, and depression were administered over eight years to 500 randomly selected adult NA men and women ages 18–64. Data were analyzed using GEE analyses taking into consideration the year of study, demographic factors, and instrumentation in examining the association between dietary exposure and outcomes. Findings indicated a significant but small decline in total recall memory within the context of otherwise stable clerical speed and accuracy and depression scores. There is reason to believe that a continuum of memory difficulties may be associated with DA exposure, rather than a unitary ASP syndrome.

scholarly journals Characterization of the Domoic Acid Uptake Mechanism of the Mussel (Mytilus galloprovincialis) Digestive Gland

Toxins ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 458
Author(s):  
Juan Blanco ◽  
Carmen Mariño ◽  
Helena Martín ◽  
Gonzalo Álvarez ◽  
Araceli E. Rossignoli
Keyword(s):  
Digestive Gland ◽  
Domoic Acid ◽  
Mytilus Galloprovincialis ◽  
Metabolic Inhibitor ◽  
Acid Uptake ◽  
Free Medium ◽  
Shellfish Poisoning ◽  
Uptake Mechanism ◽  
Amnesic Shellfish Poisoning

Cultures of the mussel Mytilus galloprovincialis are frequently affected by accumulation of the amnesic shellfish poisoning toxin domoic acid (DA). This species is characterized by a fast uptake and release of the toxin. In this work, the main characteristics of the uptake mechanism have been studied by incubation of digestive gland thin slices in media with different composition and DA concentration. DA uptake seems to follow Michaelis–Menten kinetics, with a very high estimated KM (1722 µg DA mL−1) and a Vmax of 71.9 µg DA g−1 h−1, which is similar to those found for other amino acids in invertebrates. Replacement of NaCl from the incubation media by Cl-choline (Na+-free medium) did not significantly reduce the uptake, but replacement by sorbitol (Na+-free and Cl−-depleted medium) did. A new experiment replacing all chlorides with their equivalent gluconates (Na+- and Cl−-free medium) showed an important reduction in the uptake that should be attributed to the absence of chloride, pointing to a Na+-independent, Cl− (or anion-) dependent transporter. In media with Na+ and Cl−, neither decreasing the pH nor adding cyanide (a metabolic inhibitor) had significant effect on DA uptake, suggesting that the transport mechanism is not H+- or ATP-dependent. In a chloride depleted medium, lowering pH or adding CN increased the uptake, suggesting that other anions could, at least partially, substitute chloride.

scholarly journals Monitoring the presence of domoic acid in the production areas of bivalve molluscs

2016 ◽  
Vol 5 (4) ◽  
Author(s):  
Rachele Rossi ◽  
Olga Arace ◽  
Maria Giovanna Buonomo ◽  
Daniela Capozzo ◽  
Vincenzo Castellano ◽  
...  
Keyword(s):  
Health Care ◽  
Experimental Method ◽  
Domoic Acid ◽  
Food Poisoning ◽  
Chemical Compounds ◽  
Official Method ◽  
Shellfish Poisoning ◽  
Bivalve Molluscs ◽  
Amnesic Shellfish Poisoning ◽  
Production Areas

Algal biotoxins, chemical compounds produced by some microscopic algae, constitute the <em>phytoplankton</em>. The mussels, feeding on phytoplankton, can accumulate these compounds to become themselves toxic. There have been several cases of food poisoning by consumption of contaminated shellfish. Such food poisoning have pushed our health care system to provide monitoring of shellfish in the framework of the monitoring plans carried out by AASSLL. In this paper we report the results obtained monitoring the presence of ASP (Amnesic Shellfish Poisoning) biotoxins, like domoic acid (DA) and its isomers, produced by <em>Pseudonitzschia</em> algae. The analysis were carried out by using both the HPLC-UV official method and an experimental method performed with a Time of Flight mass spectrometer (ESI-TOF). The 100% of samples analysed by the official method have always been below the limits of sensitivity (except one sample), the 65% of samples analysed by ESI-TOF, showed the presence of domoic acid.

scholarly journals High-Throughput Analysis of Amnesic Shellfish Poisoning Toxins in Shellfish by Ultra-Performance Rapid Resolution LC-MS/MS

2011 ◽  
Vol 94 (2) ◽  
pp. 555-564 ◽  
Author(s):  
Pablo De La Iglesia ◽  
Esther Barber ◽  
Gemma Giménez ◽  
María Luisa Rodríguez-Velasco ◽  
Adriano Villar-González ◽  
...  
Keyword(s):  
Domoic Acid ◽  
Proficiency Test ◽  
Direct Analysis ◽  
Z Score ◽  
Shellfish Poisoning ◽  
High Throughput Analysis ◽  
Fast Analysis ◽  
Rapid Resolution ◽  
Crude Extracts ◽  
Amnesic Shellfish Poisoning

Abstract The application of ultra-performance rapid resolution LC on a 1.8 m particle-size column coupled with tandem MS (RRLC-MS/MS) is described for the analysis of amnesic shellfish poisoning (ASP) toxins in shellfish. Complete resolution among domoic acid (DA) and the isomers was achieved in less than 3 min. The method was intralaboratory validated for direct analysis of crude extracts without further cleanup. It showed LODs ranging from 0.05 to 0.09 mg/kg and a working range that complied with the current regulatory level for DA of 20 mg/kg, and with the level of 4.5 mg/kg recently proposed by the European Food Safety Authority. Confirmatory capabilities were demonstrated according to the Commission Decision 2002/657/EC criteria. The results obtained by RRLC-MS/MS agreed with those provided by the reference LC-UV method, both intralaboratory for the analysis of blind samples (R2 0.9751) and interlaboratory through participation in the proficiency test for ASP toxins during 2009 (z-score 0.962 and 0.177 for low- and high-contaminated samples, respectively). RRLC-MS/MS provided fast analysis and additional confirmatory capabilities for direct analysis of crude extracts while the performance and reliability of the results were maintained, even in very complex matrixes.

Rapid Extraction and Cleanup for Liquid Chromatographic Determination of Domoic Acid in Unsalted Seafood

1995 ◽  
Vol 78 (2) ◽  
pp. 543-554 ◽  
Author(s):  
Michael A Quilliam ◽  
Mie Xie ◽  
William R Hardstaff
Keyword(s):  
Domoic Acid ◽  
Dynamic Range ◽  
Solid Phase ◽  
Chromatographic Determination ◽  
Single Step ◽  
Shellfish Poisoning ◽  
Rapid Extraction ◽  
Liquid Chromatographic Determination ◽  
Liquid Chromatographic

Abstract Domoic acid is the toxin responsible for incidents of amnesic shellfish poisoning. A rapid extraction and cleanup for the liquid chromatographic determination of domoic acid in unsalted seafood is reported. The method uses a single-step extraction with 50% aqueous methanol and a selective cleanup and preconcentration with strong-anion exchange, solid-phase extraction. Determination is performed by liquid chromatography with ultraviolet absorbance detection. The detection limit was 20–30 ng/g. Recoveries of 93% were achieved from 0.2 to 20 μg/g in mussel tissues. The method gave a precision of less than 3% for concentrations greater than 2 μg/g and less than 6% at 0.2 μg/g. A linear dynamic range of 104 can be achieved. The method was successfully applied to a variety of seafood products, including mussels, razor clams, crabs, and anchovies.

scholarly journals Microbiological and Marine Biotoxins Contamination of Raw Bivalve Molluscs Commercially Available in Poland

2012 ◽  
Vol 56 (4) ◽  
pp. 563-568 ◽  
Author(s):  
Remigiusz Pomykała ◽  
Mirosław Michalski ◽  
Artur Jóźwik ◽  
Jacek Osek
Keyword(s):  
Vibrio Parahaemolyticus ◽  
Paralytic Shellfish Poisoning ◽  
Shellfish Poisoning ◽  
Bivalve Molluscs ◽  
E Coli ◽  
Marine Biotoxins ◽  
Amnesic Shellfish Poisoning ◽  
Paralytic Shellfish ◽  
Coagulase Positive Staphylococci

Abstract The study was carried out on live bivalve molluscs available on Polish market. Microbiological tests were performed for the presence of Salmonella sp., Vibrio parahaemolyticus, spore-forming anaerobe bacteria, and coagulase-positive Staphylococcus sp., and for the enumeration of Escherichia coli. ELISA was used for the determination of marine biotoxins, paralytic shellfish poisoning (PSP), amnesic shellfish poisoning (ASP), and diarrhoeic shellfish poisoning (DSP). Microbiological examinations were performed according to ISO and Polish Standards. Salmonella sp. was not detected in any sample tested. Coagulase-positive staphylococci were identified in 9.0% of the samples. V. parahaemolyticus was isolated from 17.0% of mussels. Shellfish were highly contaminated by anaerobes, which were isolated from 68.0% of the samples. The number of E. coli ranged from <2.0 x 101 up to >1.8 x 104 MPN/100 g. The majority of mussels were free from the marine biotoxins tested or contained them bellow the permitted level. The analysis of microbiological and toxicological status of raw bivalve molluscs available on Polish market indicates that they are generally safe for the consumers.

scholarly journals Single-Laboratory Validation of the Biosense Direct Competitive Enzyme-Linked Immunosorbent Assay (ELISA) for Determination of Domoic Acid Toxins in Shellfish

2007 ◽  
Vol 90 (4) ◽  
pp. 1000-1010 ◽  
Author(s):  
Hans Kleivdal ◽  
Sven-Inge Kristiansen ◽  
Mona V Nilsen ◽  
Lyn Briggs
Keyword(s):  
Immunosorbent Assay ◽  
Domoic Acid ◽  
Matrix Effects ◽  
Limit Of Detection ◽  
Method Comparison ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Shellfish Poisoning ◽  
Relative Standard

Abstract Method validation was conducted for an enzyme-linked immunosorbent assay (ELISA) for the determination of domoic acid (DA) toxins, known to give amnesic shellfish poisoning (ASP) symptoms, in shellfish. The calibration curve range of the assay is approximately 10260 pg/mL, with a dynamic working range for DA toxins in shellfish from 0.01 to at least 250 mg/kg. The ASP ELISA showed no significant cross-reactivity to structural analogs, and proved to be robust to deliberate alterations of the optimal running conditions. The shellfish matrix effects observed with mussels, oysters, and scallops were eliminated by diluting shellfish extracts 1:200 prior to analysis, leading to a limit of detection at 0.003 mg/kg. Thirteen blank shellfish homogenates were spiked with certified mussel material containing DA to levels in the range of 0.125 mg DA/kg, and analyzed in quadruplicate on 3 different days. The relative standard deviation (RSD) under intra-assay repeatability conditions ranged from 6.5 to 13.1%, and under interassay repeatability conditions the RSD ranged from 5.7 to 13.4%, with a mean value of 9.3%. The recoveries ranged from 85.5 to 106.6%, with a mean recovery of 102.2%. A method comparison was conducted with liquid chromatography with ultraviolet detection, using naturally contaminated scallop samples (n = 27) with DA levels at 0244 mg/kg. The overall correlation coefficient was 0.960 and the slope of the regression was 1.218, indicating a good agreement between the methods.

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