Quantitative analysis of conventional UPLC/MS/MS of regulated and unregulated lipophilic marine biotoxins in shellfish
Arjen Gerssen 1 , Mirjam D. Klijnstra 1 , Simon Cubbon 2 and Antonietta Gledhill 2
1 Wageningen University Food Safety Institute (Waggeningen, The Netherlands)
2 Waters Corporation (Manchester, UK)
Application advantages <br> This is a way regulated lipophilic marine biotoxins in shellfish and non-regulated fast and reliable analysis. Compared to the traditional HPLC/MS/MS method, the analysis speed is increased by 4 times: (single analysis is shortened from 20 min to 5 min). The method can also be used to meet the specified detection level and can be used to replace the mouse bioassay.
Waters Solutions
ACQUITY UPLC® System
ACQUITY UPLC BEH C 18 Column
Xevo® TQ-S Mass Spectrometer
TargetLynxTM application software
QUANPEDIATM database
Keywords <br> shellfish biotoxins, lipophilic, algal toxins, mussels, clams, oysters, diarrheal shellfish poisoning, DSP
About <br> by eating contaminated shellfish biotoxins (mussels, oysters, clams, etc.) may cause serious poisoning the body, such as diarrheal shellfish poisoning (DSP). Because DSP toxins are lipophilic, they are often classified as lipophilic marine biotoxins. Marine biotoxins are naturally produced by different species of phytoplankton and are therefore also referred to as algal toxins. Lipophilic marine biotoxins exhibit versatility in terms of physicochemical properties, such as carboxylic acid, sulfonic acid, amino and imino functional groups, and thus have some complexity.
European Union (EU) legislation regulates various types of toxins and describes how these toxins are monitored in official control procedures. As shown in Figure 1A, the lipophilic biotoxin that should be monitored is Okada's soft sponge acid (OA); the algae toxins 1, 2, 3 (DTX 1, 2, 3), where DTX3 is OA, DTX 1 and 2, respectively. Ester form; scallop toxin 1, 2 (PTX1, 2); scallop toxin (YTX); 45OH scallop toxin (45OH YTX); homologous scallop toxin (homoYTX); 45OH homologous scallop toxin (45OH homoYTX); pro-polyalginic acid 1, 2 and 3 (AZA1, 2, 3) 1 .
Prior to July 2011, the official detection method for the above toxins was a biological assay based on rat oral shellfish or mice injected intraperitoneally with shellfish extracts. There are two main problems with this method: first, it is against bioethics; second, it cannot measure trace amounts of specific toxins very scientifically and reliably.
When tested by bioassay, the presence of a cyclic imine toxoid causes a physical reaction that usually kills the animal. The cyclic imine toxoid is also classified as a lipophilic marine toxin according to physicochemical properties as shown in FIG. 1B. Although these toxins are not currently regulated, the European Food Safety Authority (EFSA) notes that the toxicity data for such toxins and their presence in shellfish should be collected more.
Since July 2011, LC/MS/MS has become the official test method for lipophilic marine biotoxins in shellfish 2 . The LC/MS/MS method cited by the European Union uses a fixed extraction step to extract biotoxins, which are then separated by acidic or alkaline mobile phases by conventional LC methods and detected using tandem quadrupole MS.
The purpose of this study was to establish a routine analytical method that is faster than conventional LC methods under alkaline conditions, including the analysis of other non-regulated compounds of interest to EFSA. In this application note, we used the Waters® ACQUITY UPLC System in series with the Xevo TQ-S mass spectrometer to perform a five-minute analysis of various lipophilic marine biotoxins including some non-regular cyclic imine toxoids. . UPLC® technology reduces analysis run time without adversely affecting chromatographic peak resolution and peak quality, while the Xevo TQ-S (series quadrupole mass spectrometer) provides ultra-high sensitivity detection and is also capable of switching power Multiple reaction monitoring (MRM) transition data is acquired simultaneously in spray positive ion (ESI+) and negative ion (ESI-) modes, which is an essential step in the analysis of lipophilic marine biotoxins.
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