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

Quilliam, Michael A.; Xie, Mie; Hardstaff, W. R.

doi:10.1093/jaoac/78.2.543

Cited by 183 publications

(76 citation statements)

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“…Assays were conducted in 96-well Multi-Screen plates (Millepore , Bedford, Massachusetts), after which 25 L of Optiphase liquid scintillant were added to each well, and the well counted directly on a microplate scintillation counter (Microbeta 1450; Wallac Inc., Gaithersburg, Maryland). Selected samples were also analyzed by HPLC-UV (Quilliam et al 1995) to corroborate receptor-based DA activity, with values from both methods agreeing to within Ϯ20% (n ϭ 5). Certified reference material used to calibrate all DA measurements was the DACS-1B standard (National Research Council, Halifax, Nova Scotia, Canada).…”

Section: Methodsmentioning

confidence: 98%

PSEUDO‐NITZSCHIA SPECIES (BACILLARIOPHYCEAE) IN LOUISIANA COASTAL WATERS: MOLECULAR PROBE FIELD TRIALS, GENETIC VARIABILITY, AND DOMOIC ACID ANALYSES

Parsons

Scholin

Miller

et al. 1999

Journal of Phycology

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An 18-month field survey of the Pseudo-nitzschia population present in Louisiana coastal waters was conducted comparing species abundance estimates by novel fluorescent molecular probes (16S large subunit rDNA oligonucleotide sequences) with traditional electron and differential-interference light microscopy. While the probe and microscopic analyses agreed on the presence or absence of four common Pseudo-nitzschia species (P. multiseries (Hasle) Hasle, P. pseudodelicatissima (Hasle) Hasle, P. delicatissima (P.T. Cleve) Heiden, and P. pungens (Grunow) Hasle in 66% of the samples analyzed, the probes gave conflicting results with the microscopic methods in the remaining 34% of the samples. The majority of the discrepancies appear to be because of genetic variation within the Pseudo-nitzschia population, especially in P. pseudodelicatissima, indicating that the Monterey Bay Pseudo-nitzschia spp. may not be appropriate reference strains for distinguishing Louisiana Pseudo-nitzschia spp. Additionally, P. pseudodelicatissima has been associated with domoic acid (DA) activity in three field samples, at levels up to 22 times higher than the highest value given in 1 other published reports of DA production by this species. The contemporaneous existence of multiple strains of P. pseudodelicatissima (toxic and nontoxic) presents new challenges to the study of the ecophysiology and population dynamics of this bloom-forming species.

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Section: Methodsmentioning

confidence: 98%

PSEUDO‐NITZSCHIA SPECIES (BACILLARIOPHYCEAE) IN LOUISIANA COASTAL WATERS: MOLECULAR PROBE FIELD TRIALS, GENETIC VARIABILITY, AND DOMOIC ACID ANALYSES

Parsons

Scholin

Miller

et al. 1999

Journal of Phycology

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“…Toxin analyses of shellfish and plankton net samples were performed using the Varian HPLC‐UV‐DAD system. For determination of DA and epi‐domoic acid (epi‐DA) in shellfish, approximately 100 g of tissue was homogenized, out of which a 4 g subsample of Venus verrucosa and an 8 g subsample of other shellfish species were processed according to the protocol proposed by Quilliam et al (). Samples were cleaned using strong anion exchange (SAX) to avoid interference with tryptophan.…”

Section: Methodsmentioning

confidence: 99%

Taxonomical diversity of Pseudo‐nitzschia from the Central Adriatic Sea

Arapov

Skejić

Bužančić

et al. 2017

Phycological Research

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SUMMARY The aim of our research was to study the composition of Pseudo‐nitzschia species during a period when neurotoxin domoic acid (DA) was present in shellfish. Sampling was conducted in Kaštela Bay (Central Adriatic Sea), between November 2015 and January 2016. Concentrations of DA analyzed in various shellfish species were low, below the regulatory limit, while the highest abundance of Pseudo‐nitzschia spp. reached 1.85 × 105 cells L−1 in the surface layer, at the beginning of November. Within the temperature and salinity range recorded during the investigated period, a positive correlation of Pseudo‐nitzschia spp. abundance was recorded with temperature. Morphological analyses by scanning electron microscopy revealed the presence of five Pseudo‐nitzschia species that had already been reported in the Adriatic Sea – P. calliantha, P. delicatissima, P. fraudulenta, P. pseudodelicatissima /P. cuspidata and P. subfraudulenta – as well as an unknown Pseudo‐nitzschia sp. The composition of the Pseudo‐nitzschia assemblage changed over the investigated period. The species P. pseudodelicatissima/P.cuspidata was found throughout the entire period and the highest diversity was noticed in January, when all six observed species were recorded. These results represent the first taxonomical investigation of the genus Pseudo‐nitzschia in Kaštela Bay, as well as the first report of DA in shellfish from this area.

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“…The regulatory method for DA detection sanctioned by the Interstate Shellfish Sanitation Conference (Quilliam et al 1989a, Quilliam et al 1995) is a high performance liquid chromatography (HPLC) assay (Quilliam et al 1991. Though accurate, these analyses are generally run by centralized state facilities with results typically not available for 3-14 days after the samples are collected.…”

Section: Introductionmentioning

confidence: 99%

Rapid Enzyme-linked Immunosorbent Assay for Detection of the Algal Toxin Domoic Acid

Litaker

Stewart²,

Eberhart

et al. 2008

Journal of Shellfish Research

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Domoic acid (DA) is a potent toxin produced by bloom-forming phytoplankton in the genus Pseudo-nitzschia, which is responsible for causing amnesic shellfish poisoning (ASP) in humans. ASP symptoms include vomiting, diarrhea, and in more severe cases confusion, loss of memory, disorientation, and even coma or death. This paper describes the development and validation of a rapid, sensitive, enzyme linked immunosorbent assay test kit for detecting DA using a monoclonal antibody. The assay gives equivalent results to those obtained using standard high performance liquid chromatography, fluorenylmethoxycarbonyl high performance liquid chromatography, or liquid chromatography-mass spectrometry methods. It has a linear range from 0.1-3 ppb and was used successfully to measure DA in razor clams, mussels, scallops, and phytoplankton. The assay requires approximately 1.5 h to complete and has a standard 96-well format where each strip of eight wells is removable and can be stored at 4°C until needed. The first two wells of each strip serve as an internal control eliminating the need to run a standard curve. This allows as few as 3 or as many as 36 duplicate samples to be run at a time enabling real-time sample processing and limiting degradation of DA, which can occur during storage. There was minimal cross-reactivity in this assay with glutamine, glutamic acid, kainic acid, epi-or iso-DA. This accurate, rapid, cost-effective, assay offers environmental managers and public health officials an effective tool for monitoring DA concentrations in environment samples.

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

Cited by 183 publications

References 0 publications

PSEUDO‐NITZSCHIA SPECIES (BACILLARIOPHYCEAE) IN LOUISIANA COASTAL WATERS: MOLECULAR PROBE FIELD TRIALS, GENETIC VARIABILITY, AND DOMOIC ACID ANALYSES

PSEUDO‐NITZSCHIA SPECIES (BACILLARIOPHYCEAE) IN LOUISIANA COASTAL WATERS: MOLECULAR PROBE FIELD TRIALS, GENETIC VARIABILITY, AND DOMOIC ACID ANALYSES

Taxonomical diversity of Pseudo‐nitzschia from the Central Adriatic Sea

Rapid Enzyme-linked Immunosorbent Assay for Detection of the Algal Toxin Domoic Acid

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