Screening for the presence of lipophilic marine biotoxins in shellfish samples using the neuro-2a bioassay

Research output: Contribution to journalArticleAcademicpeer-review

4 Citations (Scopus)

Abstract

The neuro-2a bioassay is considered as one of the most promising cell-based in vitro bioassays for the broad screening of seafood products for the presence of marine biotoxins. The neuro-2a assay has been shown to detect a wide array of toxins like paralytic shellfish poisons (PSPs), ciguatoxins, and also lipophilic marine biotoxins (LMBs). However, the neuro-2a assay is rarely used for routine testing of samples due to matrix effects that, for example, lead to false positives when testing for LMBs. As a result there are only limited data on validation and evaluation of its performance on real samples. In the present study, the standard extraction procedure for LMBs was adjusted by introducing an additional clean-up step with n-hexane. Recovery losses due to this extra step were less than 10%. This wash step was a crucial addition in order to eliminate false-positive outcomes due to matrix effects. Next, the applicability of this assay was assessed by testing a broad range of shellfish samples contaminated with various LMBs, including diarrhetic shellfish toxins/poisons (DSPs). For comparison, the samples were also analysed by LC-MS/MS. Standards of all regulated LMBs were tested, including analogues of some of these toxins. The neuro-2a cells showed good sensitivity towards all compounds. Extracts of 87 samples, both blank and contaminated with various toxins, were tested. The neuro-2a outcomes were in line with those of LC-MS/MS analysis and support the applicability of this assay for the screening of samples for LMBs. However, for use in a daily routine setting, the test might be further improved and we discuss several recommended modifications which should be considered before a full validation is carried out.
LanguageEnglish
Pages351-365
JournalFood Additives & Contaminants. Pt. A, Chemistry, Analysis, Control, Exposure & Risk Assessment
Volume35
Issue number2
Early online date9 Sep 2017
DOIs
Publication statusPublished - Feb 2018

Fingerprint

Shellfish
Bioassay
shellfish
Biological Assay
Assays
Screening
bioassays
Poisons
screening
Ciguatoxins
toxins
Seafood
Testing
assays
sampling
ciguatoxin
testing
seafoods
Recovery
hexane

Keywords

  • diarrhetic shellfish poisons
  • Lipophilic marine biotoxins
  • MTT-test
  • neuro-2a cells
  • screeningfood safety

Cite this

@article{495ae47f92284795be7bf3c25f971496,
title = "Screening for the presence of lipophilic marine biotoxins in shellfish samples using the neuro-2a bioassay",
abstract = "The neuro-2a bioassay is considered as one of the most promising cell-based in vitro bioassays for the broad screening of seafood products for the presence of marine biotoxins. The neuro-2a assay has been shown to detect a wide array of toxins like paralytic shellfish poisons (PSPs), ciguatoxins, and also lipophilic marine biotoxins (LMBs). However, the neuro-2a assay is rarely used for routine testing of samples due to matrix effects that, for example, lead to false positives when testing for LMBs. As a result there are only limited data on validation and evaluation of its performance on real samples. In the present study, the standard extraction procedure for LMBs was adjusted by introducing an additional clean-up step with n-hexane. Recovery losses due to this extra step were less than 10{\%}. This wash step was a crucial addition in order to eliminate false-positive outcomes due to matrix effects. Next, the applicability of this assay was assessed by testing a broad range of shellfish samples contaminated with various LMBs, including diarrhetic shellfish toxins/poisons (DSPs). For comparison, the samples were also analysed by LC-MS/MS. Standards of all regulated LMBs were tested, including analogues of some of these toxins. The neuro-2a cells showed good sensitivity towards all compounds. Extracts of 87 samples, both blank and contaminated with various toxins, were tested. The neuro-2a outcomes were in line with those of LC-MS/MS analysis and support the applicability of this assay for the screening of samples for LMBs. However, for use in a daily routine setting, the test might be further improved and we discuss several recommended modifications which should be considered before a full validation is carried out.",
keywords = "diarrhetic shellfish poisons, Lipophilic marine biotoxins, MTT-test, neuro-2a cells, screeningfood safety",
author = "{Bodero Baeza}, Marcia and Bovee, {Toine F.H.} and S. Wang and Hoogenboom, {Ron L.A.P.} and Klijnstra, {Mirjam D.} and Liza Portier and Hendriksen, {Peter J.M.} and Arjen Gerssen",
year = "2018",
month = "2",
doi = "10.1080/19440049.2017.1368720",
language = "English",
volume = "35",
pages = "351--365",
journal = "Food Additives & Contaminants. Pt. A, Chemistry, Analysis, Control, Exposure & Risk Assessment",
issn = "1944-0049",
publisher = "Taylor & Francis",
number = "2",

}

TY - JOUR

T1 - Screening for the presence of lipophilic marine biotoxins in shellfish samples using the neuro-2a bioassay

AU - Bodero Baeza, Marcia

AU - Bovee, Toine F.H.

AU - Wang, S.

AU - Hoogenboom, Ron L.A.P.

AU - Klijnstra, Mirjam D.

AU - Portier, Liza

AU - Hendriksen, Peter J.M.

AU - Gerssen, Arjen

PY - 2018/2

Y1 - 2018/2

N2 - The neuro-2a bioassay is considered as one of the most promising cell-based in vitro bioassays for the broad screening of seafood products for the presence of marine biotoxins. The neuro-2a assay has been shown to detect a wide array of toxins like paralytic shellfish poisons (PSPs), ciguatoxins, and also lipophilic marine biotoxins (LMBs). However, the neuro-2a assay is rarely used for routine testing of samples due to matrix effects that, for example, lead to false positives when testing for LMBs. As a result there are only limited data on validation and evaluation of its performance on real samples. In the present study, the standard extraction procedure for LMBs was adjusted by introducing an additional clean-up step with n-hexane. Recovery losses due to this extra step were less than 10%. This wash step was a crucial addition in order to eliminate false-positive outcomes due to matrix effects. Next, the applicability of this assay was assessed by testing a broad range of shellfish samples contaminated with various LMBs, including diarrhetic shellfish toxins/poisons (DSPs). For comparison, the samples were also analysed by LC-MS/MS. Standards of all regulated LMBs were tested, including analogues of some of these toxins. The neuro-2a cells showed good sensitivity towards all compounds. Extracts of 87 samples, both blank and contaminated with various toxins, were tested. The neuro-2a outcomes were in line with those of LC-MS/MS analysis and support the applicability of this assay for the screening of samples for LMBs. However, for use in a daily routine setting, the test might be further improved and we discuss several recommended modifications which should be considered before a full validation is carried out.

AB - The neuro-2a bioassay is considered as one of the most promising cell-based in vitro bioassays for the broad screening of seafood products for the presence of marine biotoxins. The neuro-2a assay has been shown to detect a wide array of toxins like paralytic shellfish poisons (PSPs), ciguatoxins, and also lipophilic marine biotoxins (LMBs). However, the neuro-2a assay is rarely used for routine testing of samples due to matrix effects that, for example, lead to false positives when testing for LMBs. As a result there are only limited data on validation and evaluation of its performance on real samples. In the present study, the standard extraction procedure for LMBs was adjusted by introducing an additional clean-up step with n-hexane. Recovery losses due to this extra step were less than 10%. This wash step was a crucial addition in order to eliminate false-positive outcomes due to matrix effects. Next, the applicability of this assay was assessed by testing a broad range of shellfish samples contaminated with various LMBs, including diarrhetic shellfish toxins/poisons (DSPs). For comparison, the samples were also analysed by LC-MS/MS. Standards of all regulated LMBs were tested, including analogues of some of these toxins. The neuro-2a cells showed good sensitivity towards all compounds. Extracts of 87 samples, both blank and contaminated with various toxins, were tested. The neuro-2a outcomes were in line with those of LC-MS/MS analysis and support the applicability of this assay for the screening of samples for LMBs. However, for use in a daily routine setting, the test might be further improved and we discuss several recommended modifications which should be considered before a full validation is carried out.

KW - diarrhetic shellfish poisons

KW - Lipophilic marine biotoxins

KW - MTT-test

KW - neuro-2a cells

KW - screeningfood safety

U2 - 10.1080/19440049.2017.1368720

DO - 10.1080/19440049.2017.1368720

M3 - Article

VL - 35

SP - 351

EP - 365

JO - Food Additives & Contaminants. Pt. A, Chemistry, Analysis, Control, Exposure & Risk Assessment

T2 - Food Additives & Contaminants. Pt. A, Chemistry, Analysis, Control, Exposure & Risk Assessment

JF - Food Additives & Contaminants. Pt. A, Chemistry, Analysis, Control, Exposure & Risk Assessment

SN - 1944-0049

IS - 2

ER -