Single Laboratory Validation of A Ready-to-Use Phosphatase Inhibition Assay for Detection of Okadaic Acid Toxins

Smienk, Henry; Calvo, Dolores; Razquín, Pedro; Domı́nguez, Elena; Mata, Luis

doi:10.3390/toxins4050339

Cited by 29 publications

(18 citation statements)

References 14 publications

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“…In addition, there is a large number of articles demonstrating validation of LC-ESI-MS methods for practical analytical tasks with respect to certain validation guidelines such as FDA [68][69][70][71][72], ICH [73][74][75] Eurachem [76][77][78][79][80][81][82][83], EMA [84][85][86], IUPAC [87][88][89] or SANCO [90,91]. However, in most cases not all parameters are investigated.…”

Section: Practicalmentioning

confidence: 99%

Tutorial review on validation of liquid chromatography–mass spectrometry methods: Part I

Kruve

Rebane

Kipper

et al. 2015

Analytica Chimica Acta

246

145

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This is the part I of a tutorial review intending to give an overview of the state of the art of method validation in liquid chromatography mass spectrometry (LC-MS) and discuss specific issues that arise with MS (and MS/MS) detection in LC (as opposed to the "conventional" detectors). The Part I briefly introduces the principles of operation of LC-MS (emphasizing the aspects important from the validation point of view, in particular the ionization process and ionization suppression/enhancement); reviews the main validation guideline documents and discusses in detail the following performance parameters: selectivity/specificity/identity, ruggedness/robustness, limit of detection, limit of quantification, decision limit and detection capability. With every method performance characteristic its essence and terminology are addressed, the current status of treating it is reviewed and recommendations are given, how to determine it, specifically in the case of LC-MS methods.

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Section: Practicalmentioning

confidence: 99%

Tutorial review on validation of liquid chromatography–mass spectrometry methods: Part I

Kruve

Rebane

Kipper

et al. 2015

Analytica Chimica Acta

246

145

View full text Add to dashboard Cite

show abstract

“…PP2A inhibition fluorometric assays were also developed with a lOD of 12.8 µg OAequiv./ kg shellfish hepatopancreas (Vieytes et al, 1997), followed by further optimizations and adaptations, e.g., competitive displacement (Doskeland et al, 2000;Serres et al, 2000). Recent developments on the basis of phosphatase inhibition (single laboratory validated) suggested a lOD and a lOQ of 44 µg OAequiv./kg and 56 µg OAeequiv./kg shellfish meat, which could be confirmed with chemical analytical methods (Smienk et al, 2012). Shellfish matrix has been shown to affect the applicability of the PP2A inhibition assay when used at high concentrations.…”

Section: Standardized Reference Compoundsmentioning

confidence: 90%

A roadmap for hazard monitoring and risk assessment of marine biotoxins on the basis of chemical and biological test systems

Daneshian

2013

ALTEX

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Summary Aquatic food accounts for over 40% of global animal food products, and the potential contamination with toxins of algal origin -marine biotoxins -poses a health threat for consumers. The gold standards to assess toxins in aquatic food have traditionally been in vivo methodssingle marine biotoxins or combinations of marine biotoxins and intoxication symptomatology as well as monitoring the "success" of controls implemented during fishery and aquaculture production, processing, and retailing. For this reason, a workshop was organized in ermatingen, Switzerland by the Center for Alternatives to Animal testing -europe (CAAteurope) within the framework of the transatlantic think tank for toxicology (t 4 ).In contrast to other food products where hygiene and potential contamination with microbes or mold toxins is the prime issue, the emphasis in aquatic products (shellfish and finfish) is, beyond viral and bacterial contaminations, primarily placed on potential contamination with marine biotoxins owing to the numerous and recurring human intoxications.The gold standards to assess toxins in aquatic food have traditionally been in vivo methods, i.e., the mouse and the rat bioassays. Besides the ethical issues of in vivo bioassays there are specific difficulties, e.g., different exposure route (i.p. versus oral), low inter-species comparability, high intra-species variability, and questionable extrapolation of quantitative risk to humans, thus highlighting the need for the development of more reliable detection and quantification methods. Based on this reasoning, the european Food Safety Authority (eFSA) advocates the use of an analytical method, i.e., LC-MS (Liquid Chromatography coupled Mass Spectrometry), as a substitute for in vivo bioassays for almost all classes of marine toxins. However, although lC-MS is a very sensitive method that has the advantages of being able to detect multiple toxins in a single analysis and being good for confirming the identity of toxins, the quality of quantitative analysis is dependent on the availability of calibration standards. While many new toxin structural analogues have been detected and identified using LC-MS, this technique -as with most other methods -is not good at detecting new toxin types. When new toxin analogues are detected but accurate standards are not yet available, only an approximate quantitation is possible using estimated response factors.For the purpose of risk assessment of food, however, it is imperative to focus on the possible adverse effects, independent of whether they stem from one toxin or a combination of toxins. As toxicity is defined by functional and structural changes of biological systems, the development of a human-relevant in vitro system for appropriate risk assessment of marine biotoxins in seafood stands to reason. Moreover, poor correlation of human intoxications, of acute symptomatology and long-term adverse effects, and of predictive in vitro, in vivo, and analytical detection methodologies of marine biotoxins stems not least from a...

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“…The amount of enzyme determines the amount of analyte (OA and DTX-1) needed for the inhibition, while the quality of the enzyme ensures the amount of product formed per time unit [122]. In addition, the lack of stability or impurity of the standards used (OA and DTX-1) directly affect the quantification [123,124].…”

Section: Stages Of the Protein Phosphatase 2a Inhibition Assay For Thmentioning

confidence: 99%

“…Thus, the PP2aIA can be considered to detect toxins associated with toxins of the OA-group since its limit of detection (LOD) and limit of quantification (LOQ ) are <160 μg OA equivalents kg −1 (international maximum limit). Nevertheless, its optimal range of toxic detection is very narrow (between ≈56 and ≈96 μg OA kg −1 shellfish) due to the fact that matrices and pigments of bivalves may affect the interpretation of results [14,123]. It should be considered that samples with DTX-2 only are not able to adequately inhibit PP2a.…”

Section: Stages Of the Protein Phosphatase 2a Inhibition Assay For Thmentioning

confidence: 99%

Instrumental Methods for Detection of Lipophilic Marine Toxins in Endemic Species from Pacific Austral Fjords

Garcı́a¹,

Oyaneder-Terrazas²,

Contreras³

2019

Endemic Species

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Lipophilic marine toxins (LMTs) are a group of marine toxins which in recent years have been consistently identified in the vast majority of shellfish worldwide. One of their main characteristics is having a latitudinal variability and an assimilation/retention specific for each species. LMTs consist of four important groups: okadaic acid group (OA-group), pectenotoxin group (PTX-group), azaspiracid group (AZA-group) and yessotoxin group (YTX-group). These groups have different chemical structures, which has generated an important challenge to establish analytical techniques to identify all toxic analogues from the same toxic matrix. Likewise, in the aquatic environment, shellfish represent the best bio-indicator model that allows for the establishment of levels of toxicities related to LMTs. In this chapter, the evolution for detection of LMTs from mouse bioassay (MBA), enzymatic assays (PP2a), and analytical techniques, such as liquid chromatography tandem-mass spectrometry (LC-MS/MS), are described. These analytical advances have allowed us to determine and identify the characteristic profiles of LMTs produced by marine microalgae, including the prevalence and biotransformation of LMTs in the different endemic species. It is worth mentioning that these techniques have favoured the updating of numerous sanitary standards and the definition of the most appropriate technique for the detection of LMTs in shellfish and endemic species.

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Single Laboratory Validation of A Ready-to-Use Phosphatase Inhibition Assay for Detection of Okadaic Acid Toxins

Cited by 29 publications

References 14 publications

Tutorial review on validation of liquid chromatography–mass spectrometry methods: Part I

Tutorial review on validation of liquid chromatography–mass spectrometry methods: Part I

A roadmap for hazard monitoring and risk assessment of marine biotoxins on the basis of chemical and biological test systems

Instrumental Methods for Detection of Lipophilic Marine Toxins in Endemic Species from Pacific Austral Fjords

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