An Analytical Method of Paralytic Shellfish Toxins Using a Triple Quadrupole Mass Spectrometer

User Benefits

- It is possible to simultaneously analyze for 12 toxins (saxitoxin [STX], decarbamoylsaxitoxin [dcSTX], neosaxitoxin [NEO],decarbamoylneosaxitoxin[dcNEO], gonyautoxins [GTX1 to 4], protogonyautoxins[C1 and C2], and decarbamoylgonyautoxins [dcGTX2, dcGTX3]) using an LC-MS/MS. - Rapid analysis can be achieved within 15 minutes with a simple LC system.

Introduction

Shellfish poisoning mainly occurs when bivalves (scallops, clams, oysters, etc.) feed on toxin-containing marine dinoflagellates and these toxins accumulate in the body of the bivalve. Eating these poisonous shellfish causes four syndromes termed diarrheal shellfish poisoning, paralytic shellfish poisoning, neurotoxic shellfish poisoning, and amnesic shellfish poisoning. In Japan, the syndromes of principal concern are paralytic shellfish poisoning and diarrheal shellfish poisoning, each of which is caused by several chemicalgroups of toxins. Safety standards and regulatory limits for shellfish toxins in Japan are established based on the Food Sanitation Act (Shokuan Notice No. 0306-1, dated March 6, 2015). Regulatory levels for the toxic potency or toxic dose of the edible parts of bivalves is 4 mouse units/g or below for paralytic shellfish toxins and 0.16 mg of okadaic acid equivalent/kg or below for diarrheal shellfish toxins. This regulatory level for paralytic shellfish toxins is equivalent to 800 µg of STX·2HCl equivalent/kg, the regulatory limit cited in CODEX CXS 292-2008 and Regulation EC No. 853/2004. Paralytic shellfish toxins are currently tested for using mouse toxicity tests, but with Shokuanki Notice No. 0306-3, dated March 6, 2015, testing for diarrheal shellfish toxins is already moving to an instrumental analysis that uses LC-MS/MS. In addition, given the current status of regulations in other countries, Japan's Ministry of Agriculture, Forestry and Fisheries has cooperated with other institutions to validate instrumental methods for paralytic shellfish toxin testing and is now working on establishing guidelinesforthese methods. Internationally, paralytic shellfish toxin levels are tested by methods that include the liquid chromatography post-column oxidation and fluorescence detection method (J. AOAC Int. 2011, 94, 1154–1176), recognized by The International Ship Security Certificate (ISSC) and used by Canada, parts of the USA, Norway, and other countries, and the liquid chromatography pre-column oxidation and fluorescence detection method (AOAC 2005.06), which became an Official Reference Method of the EU in January 2019 and is used by the UK, Ireland, Portugal, New Zealand, and other countries. Due to the complexity of the HPLC systems and analytical operations needed to implement these methods, an ultra high-speed LC-MS/MS method has recently been developed that uses hydrophilic interaction liquid chromatography (HILIC).1)2) In 2018, 21 institutions participated in an international collaborative validation of this method, and the LC-MS/MS instrumental method is now considered a valid method alongside the existingHPLC method. This article describes a case study in which toxin-containing scallop samples were analyzed using the hydrophilic interaction liquid chromatography LC-MS/MSmethod.

July 28, 2022 GMT