Application of rapid test kits for the determination of paralytic shellfish poisoning (PSP) toxins in bivalve molluscs from Great Britain

Toxicon ◽  
2016 ◽  
Vol 119 ◽  
pp. 352-361 ◽  
Author(s):  
Keith Harrison ◽  
Sarah Johnson ◽  
Andrew D. Turner
Keyword(s):  
Great Britain ◽  
Rapid Test ◽  
Paralytic Shellfish Poisoning ◽  
Shellfish Poisoning ◽  
Bivalve Molluscs ◽  
Paralytic Shellfish ◽  
Psp Toxins

Assessment of a Semiquantitative Liquid Chromatography- Fluorescence Detection Method for the Determination of Paralytic Shellfish Poisoning Toxin Levels in Bivalve Molluscs from Great Britain

2014 ◽  
Vol 97 (2) ◽  
pp. 492-497 ◽  
Author(s):  
Andrew D Turner ◽  
Monika Dhanji-Rapkova ◽  
Clothilde Baker ◽  
Myriam Algoet
Keyword(s):  
Great Britain ◽  
Fluorescence Detection ◽  
Detection Method ◽  
Reference Method ◽  
Paralytic Shellfish Poisoning ◽  
Official Method ◽  
Shellfish Poisoning ◽  
Bivalve Molluscs ◽  
Paralytic Shellfish

Abstract AOAC Official Method 2005.06 precolumn oxidation LC-fluorescence detection method has been used for many years for the detection and quantitation of paralytic shellfish poisoning (PSP) toxins in bivalve molluscs. After extensive single- and multiple-laboratory validation, the method has been slowly gaining acceptance worldwide as a useful and practical tool for official control testing. In Great Britain, the method has become routine since 2008, with no requirement since then for reverting back to the bioassay reference method. Although the method has been refined to be semiautomated, faster, and more reproducible, the quantitation step can be complex and time-consuming. An alternative approach was developed to utilize the qualitative screening results for generatinga semiquantitative results assessment. Data obtained over 5 years enabled the comparison of semiquantitative and fully quantitative PSP results in over 15 000 shellfish samples comprising eight different species showed that the semiquantitative approach resulted in over-estimated paralytic shellfish toxin levels by an average factor close to two in comparison with the fully quantified levels. No temporal trends were observed in the data or relating to species type, with the exception of surf clams. The comparison suggested a semiquantitative threshold of 800 μg saxitoxin (STX) eq/kg should provide a safe limitfor the determination of samples to be forwarded to full quantitation. However, the decision was taken to halve this limit to include an additional safety factor of 2, resulting in the use of a semiquantitative threshold of 400 μg STX eq/kg. Implementation of the semiquantitative method into routine testing would result in a significant reduction in the numbers of samples requiring quantitation and have a positive impact on the overall turnaround of reported PSP results. The refined method would be appropriate for any monitoring laboratory faced with high throughput requirements.

scholarly journals Occurrence of saxitoxine (PSP marine biotoxine) in live molluscs available on Polish market

2013 ◽  
Vol 57 (4) ◽  
pp. 525-528 ◽  
Author(s):  
Mirosław M. Michalski ◽  
Katarzyna Grądziel-Krukowska ◽  
Jacek Osek
Keyword(s):  
Maximum Concentration ◽  
Paralytic Shellfish Poisoning ◽  
Shellfish Poisoning ◽  
Bivalve Molluscs ◽  
Paralytic Shellfish ◽  
Razor Clams ◽  
Psp Toxins

Abstract The study was carried out on live bivalve molluscs available on Polish market. Samples of the molluscs (n = 124) were collected from warehouses and markets. Six different species of molluscs (mussels, oysters, vongole, scallops, Japanese clams, and razor clams) were used for the determination of saxitoxine (responsible for paralytic shellfish poisoning, PSP) by ELISA. The maximum concentration of PSP toxins (756.69 μg/kg of meat) was found in scallops. The majority of tested mussels were free from the PSP toxins or contained biotoxins bellow the permitted level (800 μg/kg). The analysis of toxicological status of raw bivalve molluscs available on Polish market indicated that they are safe for consumers.

scholarly journals Assessment of MIST Alert™, a Commercial Qualitative Assay for Detection of Paralytic Shellfish Poisoning Toxins in Bivalve Molluscs

2002 ◽  
Vol 85 (3) ◽  
pp. 632-641 ◽  
Author(s):  
Fiona H Mackintosh ◽  
Susan Gallacher ◽  
Aileen M Shanks ◽  
Elizabeth A Smith
Keyword(s):  
Monitoring Program ◽  
Rapid Test ◽  
Paralytic Shellfish Poisoning ◽  
Ease Of Use ◽  
Mouse Bioassay ◽  
Shellfish Poisoning ◽  
Paralytic Shellfish ◽  
Test Kit ◽  
The Uk ◽  
Psp Toxins

Abstract A recently developed commercial rapid test kit (MIST Alert™) was assessed for determination of the presence of paralytic shellfish poisoning (PSP) toxins in shellfish. Several commercially important shellfish species obtained from the UK shellfish toxin monitoring program, containing a range of total PSP toxicities as determined by the mouse bioassay (MBA), were tested. The kit detected toxin in all samples containing the European Community tolerance level of 80 μg saxitoxin (STX) equivalents/100 g shellfish flesh as determined by the MBA. With one exception, the kit detected toxin in all samples that contained >40 μg STX equivalents/100 g according to the MBA. Among samples in which the MBA did not detect toxin, the kit disagreed in 25% of the tests, although further analysis by liquid chromatography (LC) and MBA of some samples confirmed the presence of toxins. These results suggest that MIST Alert may be suitable as an initial screen for PSP toxins as part of routine monitoring programs, thereby greatly reducing the number of MBAs. Trials were also performed by nonscientific personnel to evaluate the ease of use and interpretation of results obtained by MIST Alert. The results indicated that the kits could be readily used and accurately interpreted by individuals with no technical or scientific background.

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 Quantitative Determination of Paralytic Shellfish Poisoning Toxins in Shellfish by Using Prechromatographic Oxidation and Liquid Chromatography with Fluorescence Detection

2001 ◽  
Vol 84 (4) ◽  
pp. 1099-1108 ◽  
Author(s):  
James F Lawrence ◽  
Barbara Niedzwiadek
Keyword(s):  
Quantitative Determination ◽  
Oxidation Reaction ◽  
Solid Phase ◽  
Paralytic Shellfish Poisoning ◽  
Phase Extraction ◽  
Shellfish Poisoning ◽  
Paralytic Shellfish ◽  
Cleanup Procedure ◽  
Psp Toxins

Abstract The prechromatographic oxidation LC method developed by Lawrence [J. Assoc. Off. Anal. Chem. 74, 404–409(1991)] for the determination of paralytic shellfish poisoning (PSP) toxins has been tested for the quantitative determination of PSP toxins in shellfish. All aspects of the method were studied and modified as necessary to improve its performance for routine regulatory purposes. The chromatographic conditions were changed to shorten analysis time. The oxidation reaction was tested for repeatability and the influence of the s ample matrix on quantitation. An important part of the study was to quantitatively evaluate an ion exchange (-COOH) cleanup step using disposable solid-phase extraction cartridges that separated the PSP toxins into 3 distinct groups for quantitation, namely the C toxins, the GTX toxins, and the saxitoxin group. The cleanup step was very simple and used increasing concentrations of aqueous NaCl for elution of the toxins. The C toxins were not retained by the cartridges and thus were eluted unretained with water. The GTX toxins (GTX1 to GTX6 as well as dcGTX2 and dcGTX3) eluted from the cartridges with 0.05M NaCl while the saxitoxin group (saxitoxin, neosaxitoxin, and dcsaxitoxin) required 0.3M NaCl for elution. Each fraction was analyzed by LC after oxidation with periodate or peroxide. All of the compounds could be separated and quantitatively determined in spiked samples of mussels, clams, and oysters. The nonhydroxylated toxins could be quantitated at concentrations as low as about 0.02 μg/g (2 μg/100 g) of tissue while the hydroxylated toxins could be quantitated at concentrations as low as about 0.1 μg/g (10 μg/100 g). Average recoveries of the toxins through the complete cleanup procedure were 85%or greater for spiked extracts of oysters and clams and greater than 73%for mussels.

scholarly journals Optimization of Hydrophilic Interaction Liquid Chromatography/Mass Spectrometry and Development of Solid-Phase Extraction for the Determination of Paralytic Shellfish Poisoning Toxins

2008 ◽  
Vol 91 (6) ◽  
pp. 1372-1386 ◽  
Author(s):  
Elizabeth Turrell ◽  
Lesley Stobo ◽  
Jean-Pierre Lacaze ◽  
Sergey Piletsky ◽  
Elena Piletska
Keyword(s):  
Liquid Chromatography ◽  
Solid Phase ◽  
Paralytic Shellfish Poisoning ◽  
Phase Extraction ◽  
Liquid Chromatography Mass Spectrometry ◽  
Shellfish Poisoning ◽  
Hydrophilic Interaction ◽  
Paralytic Shellfish ◽  
Psp Toxins

Abstract The combination of hydrophilic interaction liquid chromatography (HILIC) and liquid chromatography/mass spectrometry (LC/MS) for the determination of paralytic shellfish poisoning (PSP) toxins has been proposed for use in routine monitoring of shellfish. In this study, methods for the detection of multiple PSP toxins [saxitoxin (STX), neosaxitoxin (NEO), decarbamoyl saxitoxin (dcSTX), decarbamoyl neosaxitoxin (dcNEO), gonyautoxins 15 (GTX1, GTX2, GTX3, GTX4, GTX5), decarbamoyl gonyautoxins (dcGTX2 and dcGTX3), and the N-sulfocarbamoyl C toxins (C1 and C2)] were optimized using single (MS) and triple quadrupole (MS/MS) instruments. Chromatographic separation of the toxins was achieved by using a TSK-gel Amide-80 analytical column, although superior chromatography was observed through application of a ZIC-HILIC column. Preparative procedures used to clean up shellfish extracts and concentrate PSP toxins prior to analysis were investigated. The capacity of computationally designed polymeric (CDP) materials and HILIC solid-phase extraction (SPE) cartridges to retain highly polar PSP toxins was explored. Three CDP materials and 2 HILIC cartridges were assessed for the extraction of PSP toxins from aqueous solution. Screening of the CDPs showed that all tested polymers adsorbed PSP toxins. A variety of elution procedures were examined, with dilute 0.01 acetic acid providing optimum recovery from a CDP based on 2-(trifluoromethyl)acrylic acid as the monomer. ZIC-HILIC SPE cartridges were superior to the PolyLC equivalent, with recoveries ranging from 70 to 112 (ZIC-HILIC) and 0 to 90 (PolyLC) depending on the PSP toxin. It is proposed that optimized SPE and HILIC-MS methods can be applied for the quantitative determination of PSP toxins in shellfish.

scholarly journals Quantitative Determination of Paralytic Shellfish Poisoning Toxins in Shellfish Using Prechromatographic Oxidation and Liquid Chromatography with Fluorescence Detection: Collaborative Study

2005 ◽  
Vol 88 (6) ◽  
pp. 1714-1732 ◽  
Author(s):  
James F Lawrence ◽  
Barbara Niedzwiadek ◽  
Cathie Menard ◽  
L Rojas de Astudillo ◽  
R Biré ◽  
...  
Keyword(s):  
Liquid Chromatography ◽  
Fluorescence Detection ◽  
Collaborative Study ◽  
Paralytic Shellfish Poisoning ◽  
Shellfish Poisoning ◽  
Paralytic Shellfish ◽  
Study Results ◽  
Concentration Levels ◽  
Psp Toxins

Abstract A collaborative study was conducted for the determination of paralytic shellfish poisoning (PSP) toxins in shellfish. The method used liquid chromatography with fluorescence detection after prechromatographic oxidation of the toxins with hydrogen peroxide and periodate. The PSP toxins studied were saxitoxin (STX), neosaxitoxin (NEO), gonyautoxins 2 and 3 (GTX2,3; together), gonyautoxins 1 and 4 (GTX1,4; together), decarbamoyl saxitoxin (dcSTX), B-1 (GTX5), C-1 and C-2 (C1,2; together), and C-3 and C-4 (C3,4; together). B-2 (GTX6) toxin was also included, but for qualitative identification only. Mussels, both blank and naturally contaminated, were mixed and homogenized to provide a variety of PSP toxin mixtures and concentration levels. The same procedure was followed with clams, oysters, and scallops. Twenty-one test samples in total were sent to 21 collaborators who agreed to participate in the study. Results were obtained from 18 laboratories representing 14 different countries. It is recommended that the method be adopted First Action by AOAC INTERNATIONAL.

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